Sample records for extract activates apical

  1. Protective effect of hexane and ethanol extract of piper longum L. On gentamicin-induced hair cell loss in neonatal cultures.

    PubMed

    Yadav, Mukesh Kumar; Choi, June; Song, Jae-Jun

    2014-03-01

    Gentamicin (GM) is a commonly used aminoglycoside antibiotic that generates free oxygen radicals within the inner ear, which can cause vestibulo-cochlear toxicity and permanent damage to the sensory hair cells and neurons. Piper longum L. (PL) is a well-known spice and traditional medicine in Asia and Pacific islands, which has been reported to exhibit a wide spectrum of activity, including antioxidant activity. In this study, we evaluated the effect of hexane:ethanol (2:8) PL extract (subfraction of PL [SPL] extract) on GM-induced hair cell loss in basal, middle and apical regions in a neonatal cochlea cultures. The protective effects of SPL extract were measured by phalloidin staining of cultures from postnatal day 2-3 mice with GM-induced hair cell loss. The anti-apoptosis activity of SPL extract was measured using double labeling by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and myosin-7a staining. The radical-scavenging activity of SPL extract was assessed using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. SPL extract at a concentration of 1 µg/mL significantly inhibited GM-induced hair cell loss at basal and middle region of cochlea, while 5 µg/mL was effective against apical region hair cell loss. The protective effect of SPL extract was concentration dependent and hair cells retained their stereocilia in explants treated with SPL extract prior to treatment with 0.3 mM GM. SPL extract decreased GM-induced apoptosis of hair cells as assessed by TUNEL staining. The outer hair and inner hair counts were not decreased in SPL extract treated groups in compare to GM treated explants. Additionally, SPL extract showed concentration dependent radical scavenging activity in a DPPH assay. An anti-apoptosis effect and potent radical scavenger activity of SPL extract protects from GM-induced hair cell loss at basal, middle and apical regions in neonatal cochlea cultures.

  2. Association between bacteria occurring in the apical canal system and expression of bone-resorbing mediators and matrix metalloproteinases in apical periodontitis.

    PubMed

    Takahama, A; Rôças, I N; Faustino, I S P; Alves, F R F; Azevedo, R S; Gomes, C C; Araújo-Filho, W R; Siqueira, J F

    2018-07-01

    To evaluate the association between the presence of selected bacterial species/groups in the apical root canal and expression of mediators of soft and bone tissue destruction in apical periodontitis lesions. Relationships between bacteria and some other features of apical periodontitis were also investigated. Seventeen freshly extracted teeth with pulp necrosis and apical periodontitis were included. The apical root segment was sectioned and cryopulverized; DNA was extracted and evaluated for the presence of 9 bacterial species/groups using real-time polymerase chain reaction. Lesions were processed for histopathological and immunohistochemical analyses, which targeted matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9), receptor activator of NFκB (RANK), RANK ligand (RANKL) and osteoprotegerin (OPG). Associations of the target bacteria with expression of these mediators, presence of symptoms, lesion size and histopathological diagnosis were evaluated. Data were analysed using the chi-square, Fisher's exact, Mann-Whitney and Pearson tests. P values lower than 0.05 were considered significant. All pulverized apical root samples were positive for bacteria. The most prevalent taxa were Actinobacteria (53%), Streptococcus species (35%), Fusobacterium species and Parvimonas micra (18%). The target mediators exhibited a high mean expression in the lesions (MMP-2: 82%; MMP-9: 73%; RANK: 78%; RANKL; 81%; OPG; 83%). Mean RANKL:OPG ratio was significantly higher in granulomas than cysts (P < 0.05, Mann-Whitney test). Actinobacteria were associated with granulomas, higher MMP-2 expression, lower OPG expression, and higher RANKL:OPG ratio (P < 0.05 for all, Fisher's exact test or Mann-Whitney test). No other significant associations were found. Actinobacteria may play an important role in the active phase of soft and bone tissue destruction in apical periodontitis. © 2018 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  3. Radiographic, computed tomographic, gross pathological and histological findings with suspected apical infection in 32 equine maxillary cheek teeth (2012-2015).

    PubMed

    Liuti, T; Smith, S; Dixon, P M

    2018-01-01

    Equine maxillary cheek teeth apical infections are a significant disorder because of frequent spread of infection to the supporting bones. The accuracy of computed tomographic imaging (CT) of this disorder has not been fully assessed. To compare the radiographic and CT findings in horses diagnosed with maxillary cheek teeth apical infections with pathological findings in the extracted teeth to assess the accuracy of these imaging techniques. Observational clinical study. Thirty-two maxillary cheek teeth (in 29 horses) diagnosed with apical infections by clinical, radiographic and principally by CT examinations, were extracted orally. The extracted teeth were subjected to further CT, gross pathological and histological examinations. Four normal teeth extracted from a cadaver served as controls. Pulpar and apical changes highly indicative of maxillary cheek teeth apical infection were present in all 32 teeth on CT, but in just 17/32 teeth (53%) radiographically. Gross pulpar/apical abnormalities and histological pulpar/periapical changes were present in 31/32 (97%) extracted teeth. On CT, one tooth contained small gas pockets in the apical aspect of one pulp and adjacent periodontal space, however no pathological changes were found following its extraction. The study is descriptive and is confined to a small number of cases. This study showed a 97% agreement between CT diagnosis of maxillary cheek teeth apical infection and the presence of pathological changes in the extracted teeth, confirming the diagnostic accuracy of CT compared with radiography for this disorder. © 2017 EVJ Ltd.

  4. Pro-oxidant status and matrix metalloproteinases in apical lesions and gingival crevicular fluid as potential biomarkers for asymptomatic apical periodontitis and endodontic treatment response

    PubMed Central

    2012-01-01

    Background Oxidative stress and matrix metalloproteinases -9 and -2 are involved in periodontal breakdown, whereas gingival crevicular fluid has been reported to reflect apical status. The aim of this study was to characterize oxidant balance and activity levels of MMP -2 and -9 in apical lesions and healthy periodontal ligament; and second, to determine whether potential changes in oxidant balance were reflected in gingival crevicular fluid from asymptomatic apical periodontitis (AAP)-affected teeth at baseline and after endodontic treatment. Methods Patients with clinical diagnosis of AAP and healthy volunteers having indication of tooth extraction were recruited. Apical lesions and healthy periodontal ligaments, respectively, were homogenized or processed to obtain histological tissue sections. Matrix metalloproteinase -9 and -2 levels and/or activity were analyzed by Immunowestern blot, zymography and consecutive densitometric analysis, and their tissue localization was confirmed by immunohistochemistry. A second group of patients with AAP and indication of endodontic treatment was recruited. Gingival crevicular fluid was extracted from AAP-affected teeth at baseline, after endodontic treatment and healthy contralateral teeth. Total oxidant and antioxidant status were determined in homogenized tissue and GCF samples. Statistical analysis was performed using STATA v10 software with unpaired t test, Mann-Whitney test and Spearman's correlation. Results Activity of MMP-2 and MMP-9 along with oxidant status were higher in apical lesions (p < 0.05). Total oxidant status correlated positively with matrix metalloproteinase-2 and lesion size (p < 0.05). Gingival crevicular fluid showed significantly lower levels of total antioxidant status in diseased teeth at baseline compared to controls and endodontically-treated groups. Conclusions Apical lesions display an oxidant imbalance along with increased activity of matrix metalloproteinase-2 and -9 and might contribute to AAP progression. Oxidant imbalance can also be reflected in GCF from AAP-affected teeth and was restored to normal levels after conservative endodontic treatment. These mediators might be useful as potential biomarkers for chair-side complementary diagnostic of apical status in GCF. PMID:22436166

  5. Pro-oxidant status and matrix metalloproteinases in apical lesions and gingival crevicular fluid as potential biomarkers for asymptomatic apical periodontitis and endodontic treatment response.

    PubMed

    Dezerega, Andrea; Madrid, Sonia; Mundi, Verónica; Valenzuela, María A; Garrido, Mauricio; Paredes, Rodolfo; García-Sesnich, Jocelyn; Ortega, Ana V; Gamonal, Jorge; Hernández, Marcela

    2012-03-21

    Oxidative stress and matrix metalloproteinases -9 and -2 are involved in periodontal breakdown, whereas gingival crevicular fluid has been reported to reflect apical status. The aim of this study was to characterize oxidant balance and activity levels of MMP -2 and -9 in apical lesions and healthy periodontal ligament; and second, to determine whether potential changes in oxidant balance were reflected in gingival crevicular fluid from asymptomatic apical periodontitis (AAP)-affected teeth at baseline and after endodontic treatment. Patients with clinical diagnosis of AAP and healthy volunteers having indication of tooth extraction were recruited. Apical lesions and healthy periodontal ligaments, respectively, were homogenized or processed to obtain histological tissue sections. Matrix metalloproteinase -9 and -2 levels and/or activity were analyzed by Immunowestern blot, zymography and consecutive densitometric analysis, and their tissue localization was confirmed by immunohistochemistry. A second group of patients with AAP and indication of endodontic treatment was recruited. Gingival crevicular fluid was extracted from AAP-affected teeth at baseline, after endodontic treatment and healthy contralateral teeth. Total oxidant and antioxidant status were determined in homogenized tissue and GCF samples. Statistical analysis was performed using STATA v10 software with unpaired t test, Mann-Whitney test and Spearman's correlation. Activity of MMP-2 and MMP-9 along with oxidant status were higher in apical lesions (p < 0.05). Total oxidant status correlated positively with matrix metalloproteinase-2 and lesion size (p < 0.05). Gingival crevicular fluid showed significantly lower levels of total antioxidant status in diseased teeth at baseline compared to controls and endodontically-treated groups. Apical lesions display an oxidant imbalance along with increased activity of matrix metalloproteinase-2 and -9 and might contribute to AAP progression. Oxidant imbalance can also be reflected in GCF from AAP-affected teeth and was restored to normal levels after conservative endodontic treatment. These mediators might be useful as potential biomarkers for chair-side complementary diagnostic of apical status in GCF.

  6. Correlation between calmodulin activity and gravitropic sensitivity in primary roots of maize

    NASA Technical Reports Server (NTRS)

    Stinemetz, C. L.; Kuzmanoff, K. M.; Evans, M. L.; Jarrett, H. W.

    1987-01-01

    Recent evidence indicates a role for calcium and calmodulin in the gravitropic response of primary roots of maize (Zea mays, L.). We examined this possibility by testing the relationship between calmodulin activity and gravitropic sensitivity in roots of the maize cultivars Merit and B73 x Missouri 17. Roots of the Merit cultivar require light to the gravitropically competent. The gravitropic response of the Missouri cultivar is independent of light. The occurrence of calmodulin in primary roots of these maize cultivars was tested by affinity gel chromatography followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with bovine brain calmodulin as standard. The distribution of calmodulin activity was measured using both the phosphodiesterase and NAD kinase assays for calmodulin. These assays were performed on whole tissue segments, crude extracts, and purified extracts. In light-grown seedlings of the Merit cultivar or in either dark- or light-grown seedlings of the Missouri cultivar, calmodulin activity per millimeter of root tissue was about 4-fold higher in the apical millimeter than in the subtending 3 millimeters. Calmodulin activity was very low in the apical millimeter of roots of dark-grown (gravitropically nonresponsive) seedlings of the Merit cultivar. Upon illumination, the calmodulin activity in the apical millimeter increased to a level comparable to that of light-grown seedlings and the roots became gravitropically competent. The time course of the development of gravitropic sensitivity following illumination paralleled the time course of the increase in calmodulin activity in the apical millimeter of the root. The results are consistent with the suggestion that calmodulin plays an important role in the gravitropic response of roots.

  7. Effect of four different intracanal medicaments on the apical seal of the root canal system: a dye extraction study.

    PubMed

    Tandan, Monika; Hegde, Mithra N; Hegde, Priyadarshini

    2014-01-01

    The aim was to determine the effect of four different intracanal medicaments on the apical seal of the root canal system in vitro. Fifty freshly extracted intact human permanent maxillary central incisors were collected, stored and disinfected. The root canals were prepared to a master apical size of number 50 using step back technique. Depending upon the intracanal medicament used, the teeth were divided randomly into five groups of 10 teeth each including one control group and four experimental groups. Group A: No intracanal medicament. Group B: Calcium hydroxide powder mixed with distilled water. Group C: Calcium hydroxide gutta percha points (calcium hydroxide points). Group D: 1% chlorhexidine gel (hexigel). Group E: Chlorhexidine gutta percha points (Roeko Activ Points). The medication was left in canals for 14 days. Following removal of the intracanal medicament, all the groups were obturated with lateral compaction technique. The apical leakage was then evaluated using dye extraction method with the help of a spectrophotometer. RESULTS were statistically analyzed using Kruskal-Wallis and Mann-Whitney U-test, which showed statistically significant difference among the five groups tested. It can be concluded from this study that the control group showed least amount of leakage, whereas the 1% chlorhexidine gel group showed maximum amount of leakage. Apical leakage was observed with all the experimental groups with little variations in between them. Under the parameters of this study, it can be concluded that use of intracanal medicaments during endodontic treatment has a definite impact on the apical seal of the root canal system.

  8. Assessment of apical transportation caused by nickel-titanium rotary systems with full rotation and reciprocating movements using extracted teeth and resin blocks with simulated root canals: A comparative study.

    PubMed

    Alrahabi, M; Zafar, M S

    2018-06-01

    : We compared apical transportation in the WaveOne and ProTaper Next systems, which are rotary nickel-titanium systems with reciprocating and continuous rotation movements, respectively, using manual measurements obtained from resin blocks with simulated root canals and double digital radiographs of extracted teeth. : We used 30 resin blocks with simulated root canals and 30 extracted teeth for this study. The same endodontist performed root canal shaping using the WaveOne or ProTaper Next system. We assessed apical transportation by measuring the amounts (in mm) of material lost 1 mm from the apical foramen in the resin blocks and by using double digital radiography for the extracted teeth. Significant differences between groups were assessed using t-tests. P < 0.05 was considered statistically significant. : The amount of apical transportation differed significantly between the two systems when resin blocks were used for assessment (P < 0.05), but there were no significant differences when extracted teeth were used (P < 0.05). In the current study, there was no significant difference in apical transportation between natural teeth prepared using WaveOne and those prepared using ProTaper Next. However, significant differences were observed between the two systems with resin blocks. These findings indicate that the use of resin blocks is not an accurate method for apical transportation evaluation.

  9. Comparative evaluation of total RNA extraction methods in Theobroma cacao using shoot apical meristems.

    PubMed

    Silva, D V; Branco, S M J; Holanda, I S A; Royaert, S; Motamayor, J C; Marelli, J P; Corrêa, R X

    2016-03-04

    Theobroma cacao is a species of great economic importance with its beans used for chocolate production. The tree has been a target of various molecular studies. It contains many polyphenols, which complicate the extraction of nucleic acids with the extraction protocols requiring a large amount of plant material. These issues, therefore, necessitate the optimization of the protocols. The aim of the present study was to evaluate different methods for extraction of total RNA from shoot apical meristems of T. cacao 'CCN 51' and to assess the influence of storage conditions for the meristems on the extraction. The study also aimed to identify the most efficient protocol for RNA extraction using a small amount of plant material. Four different protocols were evaluated for RNA extraction using one shoot apical meristem per sample. Among these protocols, one that was more efficient was then tested to extract RNA using four different numbers of shoot apical meristems, subjected to three different storage conditions. The best protocol was tested for cDNA amplification using reverse transcription-polymerase chain reaction; the cDNA quality was determined to be satisfactory for molecular analyses. The study revealed that with the best RNA extraction protocol, one shoot apical meristem was sufficient for extraction of high-quality total RNA. The results obtained might enable advances in genetic analyses and molecular studies using reduced amount of plant material.

  10. Apical and root canal space sealing abilities of resin and glass ionomer-based root canal obturation systems.

    PubMed

    Royer, Kinga; Liu, Xue Jun; Zhu, Qiang; Malmstrom, Hans; Ren, Yan-Fang

    2013-01-01

    To investigate the apical sealing ability of glass ionomer and resin-based root canal obturation systems in comparison to a conventional vertical compaction of warm guttapercha. Forty-five extracted human teeth were randomly assigned into 3 groups of 15 each: a resin-based (EndoRez), a glass ionomer-based (Activ GP), and a conventional gutta-percha plus pulp sealer obturation system (GP/EWT). Apical and root canal space sealing abilities were assessed on five cross-sections 1.0 mm apart starting from the apex. Cross-section images were analysed using a focus-variation 3D scanning microscope and unsealed space was calculated as the percentage of total root canal space occupied by voids and debris. EndoRez had significantly higher rate of apical leakage and deeper dye penetration as compared to GP/EWT and Activ GP. EndoRez group had also more voids and debris (22.5%) in the root canal spaces as compared to GP/EWT (10.5%) and Activ GP (10.8%). Apical leakages occurred not only along the root canal walls, but also along the gutta-percha cones with EndoRez as a result of significant polymerisation shrinkage of the resin sealer. Resin-based EndoRez did not form an adequate apical seal of filled root canals. Glass ionomer-based Activ GP was comparable to a vertical compaction of warm guttapercha plus EWT sealer in sealing root canal spaces.

  11. Evaluation of Apical Vapor Lock Formation and comparative Evaluation of its Elimination using Three different Techniques: An in vitro Study.

    PubMed

    Agarwal, Anand; Deore, Rahul B; Rudagi, Kavitarani; Nanda, Zinnie; Baig, Mirza Osman; Fareez, Md Adil

    2017-09-01

    The aim of this study was (i) to evaluate the formation of air bubbles in the apical region of root canal (apical vapor lock) during syringe irrigation, using cone beam computed tomography (CBCT) and (ii) comparative evaluation of the elimination of an established vapor lock by EndoActivator, ultrasonics, and manual dynamic agitation (MDA), using CBCT. A total of 60 extracted human single-rooted teeth were equally divided into three groups of 20 teeth each. The samples were decoronated 17 mm from the apex, cleaned, and shaped to size F4 Protaper using 3% sodium hypochlorite. Samples were irrigated with 3% sodium hypochlorite + cesium chloride radiopaque dye, and preoperative CBCT images were obtained. After formation of apical vapor lock in the scanned teeth, EndoActivator (group I), passive ultrasonic irrigation (group II), and MDA with K-file (group III) were performed and the teeth were again placed in CBCT scanner and results analyzed using the chi-square test. The apical vapor lock was formed in all the samples. Out of the 20 teeth in each group, the apical vapor lock was eliminated in 18 samples of EndoActivator group (90%), 16 samples of ultrasonic group (80%), while it was eliminated in 10 samples by MDA (50%). It is concluded that (1) apical vapor lock is consistently formed during endodontic irrigation in closed canal systems and (2) sonic activation performs better than the ultrasonics and MDA in eliminating the apical vapor lock, with statistically significant difference between all the three groups (p < 0.05). The results suggest that the apical vapor lock (dead water zone) is consistently formed during routine endodontic irrigation which impedes irrigant penetration till the working length, thereby leading to inefficient debridement. Hence, to eliminate this vapor lock, techniques, such as sonics or ultrasonics should be used along with the irrigant after shaping and cleaning of the root canal.

  12. Evaluation of 4 Different Irrigating Systems for Apical Extrusion of Sodium Hypochlorite.

    PubMed

    Yost, Ross A; Bergeron, Brian E; Kirkpatrick, Timothy C; Roberts, Mark D; Roberts, Howard W; Himel, Van T; Sabey, Kent A

    2015-09-01

    The aim of this study was to evaluate NaOCl apical extrusion by using negative apical pressure (EndoVac), sonic agitation (EndoActivator), side-vented needle (Max-i-Probe), and photon induced photoacoustic streaming (PIPS 10 mJ and PIPS 20 mJ) laser irrigation in an in vitro gel model. Extracted mandibular and maxillary central incisors (n = 18) were prepared to size 35/.04 and 55/.04, respectively. Teeth were mounted in transparent containers with clear acrylic and suspended in a color-changing pH-sensitive gel, creating a closed system. By using a crossover design, each tooth was sequentially irrigated by using 6% NaOCl with each device following manufacturers' recommendations. Each tooth served as its own control. Pre-irrigation and post-irrigation buccal and proximal view photographs served to measure the longest distance of extrusion and were analyzed with ImageJ software. Mean results were analyzed by using Kruskal-Wallis and Dunn post hoc test (P < .05). There were no significant differences between EndoVac, EndoActivator, and the passive extrusion groups. The EndoVac and EndoActivator groups produced significantly less extrusion than PIPS irrigation. Max-i-Probe extrusion results were more variable than those of EndoActivator but had no significant difference. Across all irrigation systems, there were no significant differences with respect to apical preparation size. Under the in vitro conditions of this study, no difference was found between the 10 mJ and 20 mJ PIPS laser groups. EndoVac demonstrated significantly less potential for apical extrusion than PIPS and Max-i-Probe, whereas apical preparation size did not significantly affect extrusion of irrigant. The potential for apical extrusion of endodontic irrigants should be a consideration when selecting a system for final irrigation. Published by Elsevier Inc.

  13. Bone resorptive activity in symptomatic and asymptomatic apical lesions of endodontic origin.

    PubMed

    Salinas-Muñoz, M; Garrido-Flores, M; Baeza, M; Huamán-Chipana, P; García-Sesnich, J; Bologna, R; Vernal, R; Hernández, M

    2017-11-01

    The aim of this study is to assess the levels and diagnostic accuracy of a set of bone resorption biomarkers, including TRAP-5, RANKL, and OPG in symptomatic and asymptomatic apical lesions and controls. Apical tissues from symptomatic and asymptomatic apical periodontitis patients and periodontal ligaments from healthy teeth extracted for orthodontic reasons were processed for tissue homogenization and the levels of TRAP-5, RANKL, and OPG were determined by multiplex assay. Marker levels were analyzed by Kruskal Wallis test, and diagnostic accuracy was analyzed with ROC curves. Higher levels of RANKL, OPG, and RANKL/OPG ratio were determined in both types of apical lesions compared to healthy periodontal ligament, whereas higher TRAP-5 levels were found only in symptomatic apical lesions (p < 0.05). OPG, RANKL, and RANKL/OPG ratio showed diagnostic potential to identify apical lesions versus healthy controls (AUC = 0.69, p < 0.05); while TRAP-5 showed a potential to discriminate symptomatic versus asymptomatic apical periodontitis (AUC = 0.71, p < 0.05) and healthy controls (AUC = 0.83, p < 0.05). Apical lesions showed higher RANKL and OPG levels than healthy tissues. TRAP-5 levels were the highest in symptomatic apical lesions, suggesting that these represent a progressive state, and showed diagnostic potential. Clinically symptomatic apical periodontitis might represent biologically progressive apical lesions based on TRAP5 levels. TRAP5 has diagnostic potential to identify these lesions, representing a candidate prognostic biomarker.

  14. SEM observations of resected root canal ends following apicoectomy.

    PubMed

    Furusawa, Masahiro; Asai, Yasuhiro

    2002-02-01

    The purpose of this study was to examine the apical foramen of root apices extracted during apicotomies. A total of 25 teeth extracted from 25 patients admitted to the Department of Conservative Dentistry at Tokyo Dental College's Chiba Hospital were used for the study. All patients were between 22 to 56 years of age at the time of the study, and each of the 25 cases was determined clinically on radiographs to be chronic apical suppurative periodontitis. Microsurgery was performed on all cases, and the extracted root apices were then observed using SEM. The results demonstrated a wide opening, greater than 350 microns as measured along the major axis, of the apical foramen in 80% of the cases. Various characteristics indicative of resorption were observed around the apical foramen. These features included those believed to have been caused by overinstrumentation during root canal treatment as well as irregularly shaped areas presumed to be apical lesions that had enlarged and eroded. We observed a high frequency of manifestations of cementum resorption surrounding the root apices of teeth with apical lesions. Furthermore, we concluded that in the majority of cases in the present study, due to the fact that the apical foramen exceeded normal opening dimensions as a result of overinstrumentation during root canal treatment or resorption around the root apex, prolongation of the lesions had occurred in response to direct contact of microbial infectious matter and tissues surrounding the root apex over a large area. The above finding suggested that, in cases in which the apical foramen is destroyed through overinstrumentation larger than #35 or in which the apical foramen opens up to dimensions greater than 350 microns due to pathologic resorption, surgical intervention may be indicated. On the other hand, in 64% of the cases, an accessory canal was observed in the root apical lesion. Based on this observation, the presence of an accessory canal in the root apex may contribute to some degree to the prolongation of the lesion.

  15. Apical sorting of a voltage- and Ca2+-activated K+ channel α-subunit in Madin-Darby canine kidney cells is independent of N-glycosylation

    PubMed Central

    Bravo-Zehnder, Marcela; Orio, Patricio; Norambuena, Andrés; Wallner, Martin; Meera, Pratap; Toro, Ligia; Latorre, Ramón; González, Alfonso

    2000-01-01

    The voltage- and Ca2+-activated K+ (KV,Ca) channel is expressed in a variety of polarized epithelial cells seemingly displaying a tissue-dependent apical-to-basolateral regionalization, as revealed by electrophysiology. Using domain-specific biotinylation and immunofluorescence we show that the human channel KV,Ca α-subunit (human Slowpoke channel, hSlo) is predominantly found in the apical plasma membrane domain of permanently transfected Madin-Darby canine kidney cells. Both the wild-type and a mutant hSlo protein lacking its only potential N-glycosylation site were efficiently transported to the cell surface and concentrated in the apical domain even when they were overexpressed to levels 200- to 300-fold higher than the density of intrinsic Slo channels. Furthermore, tunicamycin treatment did not prevent apical segregation of hSlo, indicating that endogenous glycosylated proteins (e.g., KV,Ca β-subunits) were not required. hSlo seems to display properties for lipid-raft targeting, as judged by its buoyant distribution in sucrose gradients after extraction with either detergent or sodium carbonate. The evidence indicates that the hSlo protein possesses intrinsic information for transport to the apical cell surface through a mechanism that may involve association with lipid rafts and that is independent of glycosylation of the channel itself or an associated protein. Thus, this particular polytopic model protein shows that glycosylation-independent apical pathways exist for endogenous membrane proteins in Madin-Darby canine kidney cells. PMID:11069304

  16. Inhibition of vinblastine efflux mediated by P-glycoprotein by grapefruit juice components in caco-2 cells.

    PubMed

    Takanaga, H; Ohnishi, A; Matsuo, H; Sawada, Y

    1998-10-01

    We investigated the effect of components in grapefruit juice (GFJ) on the transport of vinblastine, a substrate of P-glycoprotein (P-gp), across Caco-2 cells. The apical to basolateral flux of [3H]vinblastine was increased in the presence of GFJ extracts. The steady-state uptake of [3H]vinblastine from the apical side was significantly increased in the presence of GFJ in a dose-dependent manner within the range of 2.5 to 50% (v/v) of GFJ. Although naringin and naringenin reduced apical efflux of [3H]vinblastine at the concentration present in GFJ and increased steady-state uptake from the apical side to 124 and 240%, respectively, the observed effect of naringin was not enough to account for the effect of GFJ and naringenin is not naturally present in GFJ. To investigate the effective components in GFJ, we examined the inhibitory effect of several organic solvent extracts of GFJ on the transport of [3H]vinblastine in Caco-2 cells. Organic solvent extracts of GFJ enhanced the apical to basolateral transcellular transport and inhibited the apical efflux. The permeability coefficient of apical to basolateral transport of [3H]vinblastine increased in the order of the ethyl acetate>diethyl ether>methylene chloride extracts of GFJ. Since the extracted amount of naringenin by ethyl acetate was less than that with the other organic solvents, the primary inhibitor in GFJ is suggested to be different from this flavonoid. The present study demonstrated the existence of inhibitory components in GFJ for the P-gp function in Caco-2 cells, which are distinct from known components such as naringin or naringenin.

  17. Exogenous Methyl Jasmonate Treatment Increases Glucosinolate Biosynthesis and Quinone Reductase Activity in Kale Leaf Tissue

    PubMed Central

    Ku, Kang-Mo; Jeffery, Elizabeth H.; Juvik, John A.

    2014-01-01

    Methyl jasmonate (MeJA) spray treatments were applied to the kale varieties ‘Dwarf Blue Curled Vates’ and ‘Red Winter’ in replicated field plantings in 2010 and 2011 to investigate alteration of glucosinolate (GS) composition in harvested leaf tissue. Aqueous solutions of 250 µM MeJA were sprayed to saturation on aerial plant tissues four days prior to harvest at commercial maturity. The MeJA treatment significantly increased gluconasturtiin (56%), glucobrassicin (98%), and neoglucobrassicin (150%) concentrations in the apical leaf tissue of these genotypes over two seasons. Induction of quinone reductase (QR) activity, a biomarker for anti-carcinogenesis, was significantly increased by the extracts from the leaf tissue of these two cultivars. Extracts of apical leaf tissues had greater MeJA mediated increases in phenolics, glucosinolate concentrations, GS hydrolysis products, and QR activity than extracts from basal leaf tissue samples. The concentration of the hydrolysis product of glucoraphanin, sulforphane was significantly increased in apical leaf tissue of the cultivar ‘Red Winter’ in both 2010 and 2011. There was interaction between exogenous MeJA treatment and environmental conditions to induce endogenous JA. Correlation analysis revealed that indole-3-carbanol (I3C) generated from the hydrolysis of glucobrassicin significantly correlated with QR activity (r = 0.800, P<0.001). Concentrations required to double the specific QR activity (CD values) of I3C was calculated at 230 µM, which is considerably weaker at induction than other isothiocyanates like sulforphane. To confirm relationships between GS hydrolysis products and QR activity, a range of concentrations of MeJA sprays were applied to kale leaf tissues of both cultivars in 2011. Correlation analysis of these results indicated that sulforaphane, NI3C, neoascorbigen, I3C, and diindolylmethane were all significantly correlated with QR activity. Thus, increased QR activity may be due to combined increases in phenolics (quercetin and kaempferol) and GS hydrolysis product concentrations rather than by individual products alone. PMID:25084454

  18. Immediate Implant Placement in Sockets with Asymptomatic Apical Periodontitis.

    PubMed

    Crespi, Roberto; Capparé, Paolo; Crespi, Giovanni; Lo Giudice, Giuseppe; Gastaldi, Giorgio; Gherlone, Enrico

    2017-02-01

    The purpose of the present study was to evaluate if the presence of granulation tissue in asymptomatic apical periodontitis compromised immediate implant placement. Patients requiring extraction of one tooth (maxillary and mandibular incisive, canine or premolar) with asymptomatic apical periodontitis, were recruited for this prospective study. They were randomly scheduled into two groups: in first group (A) including 30 teeth, reactive soft tissue was debrided before implant placement, and in second group (B) including 30 teeth, reactive soft tissue was left in the apical lesion. Implants were positioned immediately after tooth extraction, and were loaded after 3 months in both groups. Cone beam computed tomography was performed before tooth extraction and at 1-year follow-up to evaluate the radiolucency around the root apex and the implant, bucco-lingual bone levels were also checked. Sixty patients were included in this study. Sixty implants were placed immediately after tooth extraction and, at 1-year follow-up, a survival rate of 100% was reported. After one year both groups showed absence of radiolucent zone at the apical region of implants. All fresh sockets presented a buccal-palatal bone reduction in both groups after one year, even if not statistically significant differences were found between baseline bone levels and within groups. Within the limitations of the present study, the immediate placement of implants into the extraction sockets with asymptomatic apical periodontitis, in presence of primary stability, did not lead to an increased rate of complications and rendered an equally favorable type of tissue integration. © 2016 Wiley Periodicals, Inc.

  19. Allergenic proteases cleave the chemokine CX3CL1 directly from the surface of airway epithelium and augment the effect of rhinovirus.

    PubMed

    Loxham, M; Smart, D E; Bedke, N J; Smithers, N P; Filippi, I; Blume, C; Swindle, E J; Tariq, K; Howarth, P H; Holgate, S T; Davies, D E

    2018-03-01

    CX3CL1 has been implicated in allergen-induced airway CD4 + T-lymphocyte recruitment in asthma. As epidemiological evidence supports a viral infection-allergen synergy in asthma exacerbations, we postulated that rhinovirus (RV) infection in the presence of allergen augments epithelial CX3CL1 release. Fully differentiated primary bronchial epithelial cultures were pretreated apically with house dust mite (HDM) extract and infected with rhinovirus-16 (RV16). CX3CL1 was measured by enzyme-linked immunosorbent assay and western blotting, and shedding mechanisms assessed using inhibitors, protease-activated receptor-2 (PAR-2) agonist, and recombinant CX3CL1-expressing HEK293T cells. Basolateral CX3CL1 release was unaffected by HDM but stimulated by RV16; inhibition by fluticasone or GM6001 implicated nuclear factor-κB and ADAM (A Disintegrin and Metalloproteinase) sheddases. Conversely, apical CX3CL1 shedding was stimulated by HDM and augmented by RV16. Although fluticasone or GM6001 reduced RV16+HDM-induced apical CX3CL1 release, heat inactivation or cysteine protease inhibition completely blocked CX3CL1 shedding. The HDM effect was via enzymatic cleavage of CX3CL1, not PAR-2 activation, yielding a product mitogenic for smooth muscle cells. Extracts of Alternaria fungus caused similar CX3CL1 shedding. We have identified a novel mechanism whereby allergenic proteases cleave CX3CL1 from the apical epithelial surface to yield a biologically active product. RV16 infection augmented HDM-induced CX3CL1 shedding-this may contribute to synergy between allergen exposure and RV infection in triggering asthma exacerbations and airway remodeling.

  20. Differential subcellular membrane recruitment of Src may specify its downstream signalling

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Diesbach, Philippe de; Medts, Thierry; Carpentier, Sarah

    2008-04-15

    Most Src family members are diacylated and constitutively associate with membrane 'lipid rafts' that coordinate signalling. Whether the monoacylated Src, frequently hyperactive in carcinomas, also localizes at 'rafts' remains controversial. Using polarized MDCK cells expressing the thermosensitive v-Src/tsLA31 variant, we here addressed how Src tyrosine-kinase activation may impact on its (i) membrane recruitment, in particular to 'lipid rafts'; (ii) subcellular localization; and (iii) signalling. The kinetics of Src-kinase thermoactivation correlated with its recruitment from the cytosol to sedimentable membranes where Src largely resisted solubilisation by non-ionic detergents at 4 deg. C and floated into sucrose density gradients like caveolin-1 andmore » flotillin-2, i.e. 'lipid rafts'. By immunofluorescence, activated Src showed a dual localization, at apical endosomes/macropinosomes and at the apical plasma membrane. The plasma membrane Src pool did not colocalize with caveolin-1 and flotillin-2, but extensively overlapped GM1 labelling by cholera toxin. Severe ({approx} 70%) cholesterol extraction with methyl-{beta}-cyclodextrin (M{beta}CD) did not abolish 'rafts' floatation, but strongly decreased Src association with floating 'rafts' and abolished its localization at the apical plasma membrane. Src activation independently activated first the MAP-kinase - ERK1/2 pathway, then the PI3-kinase - Akt pathway. MAP-kinase - ERK1/2 activation was insensitive to M{beta}CD, which suppressed Akt phosphorylation and apical endocytosis induced by Src, both depending on the PI3-kinase pathway. We therefore suggest that activated Src is recruited at two membrane compartments, allowing differential signalling, first via ERK1/2 at 'non-raft' domains on endosomes, then via PI3-kinase-Akt on a distinct set of 'rafts' at the apical plasma membrane. Whether this model is applicable to c-Src remains to be examined.« less

  1. Apical root canal microbiota as determined by reverse-capture checkerboard analysis of cryogenically ground root samples from teeth with apical periodontitis.

    PubMed

    Rôças, Isabela N; Alves, Flávio R F; Santos, Adriana L; Rosado, Alexandre S; Siqueira, José F

    2010-10-01

    Bacteria located in the apical root canal system potentially participate in the pathogenesis of apical periodontitis. Detection and identification of apical bacteria can be compromised because of limitations in conventional sampling and identification procedures. This study identified several bacterial taxa in the apical and middle/coronal segments of primarily infected root canal system by using pulverized root segments and a culture-independent molecular method. Seventeen extracted teeth with attached apical periodontitis lesions were sectioned to obtain 2 root fragments (apical and middle/coronal segments). Root fragments were cryogenically ground, and DNA was extracted from samples. After multiple displacement amplification, DNA from samples was used as template in a reverse-capture checkerboard hybridization assay targeting 28 bacterial taxa. Bacterial DNA was detected in all samples. The most prevalent taxa in the apical root canal system were Olsenella uli (76.5%), Prevotella baroniae (71%), Porphyromonas endodontalis (65%), Fusobacterium nucleatum (53%), and Tannerella forsythia (47%). O. uli, P. endodontalis, and Propionibacterium acnes were as frequently detected in apical samples as they were in middle/coronal samples. P. baroniae, T. forsythia, and F. nucleatum were found more frequently in the apical part of the canal as compared with matched coronal segments. Streptococcus species were more prevalent in middle/coronal samples. The median and mean of shared bacterial taxa between matched apical and middle/coronal segments were 27% and 41%, respectively. Several candidate endodontic pathogens were very prevalent in the apical root canal system. The apical microbiota was usually complex and differed in species composition when compared with the microbiota of middle/coronal samples from the same tooth. Copyright © 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  2. Efficacy of laser-driven irrigation versus ultrasonic in removing an airlock from the apical third of a narrow root canal.

    PubMed

    Peeters, Harry Huiz; Gutknecht, Norbert

    2014-08-01

    The purpose of the study was to test the hypothesis that air entrapment occurs in the apical third of a root canal during irrigation. A second objective was to test the null hypothesis that there is no difference between laser-driven irrigation (an erbium, chromium:yttrium-scandium-gallium-garnet laser) and passive ultrasonic irrigation in removing an airlock from the apical third. One hundred twenty extracted human teeth with single narrow root canals were randomised into two experimental groups (n = 40) and two control groups (n = 20). The specimens were shaped using hand instruments up to a size 30/0.02 file. The teeth were irrigated with a mixture of saline, radiopaque contrast and ink in solution. In the passive ultrasonic irrigation group, the irrigant was activated with an ultrasonic device for 60 s. In the laser group, the irrigant was activated with a laser for 60 s. It was concluded that if the insertion of irrigation needle is shorter than the working length, air entrapment may develop in the apical third, but the use of laser-driven irrigation is completely effective in removing it. © 2013 The Authors. Australian Endodontic Journal © 2013 Australian Society of Endodontology.

  3. Monocyte chemotactic protein-3: possible involvement in apical periodontitis chemotaxis.

    PubMed

    Dezerega, A; Osorio, C; Mardones, J; Mundi, V; Dutzan, N; Franco, M; Gamonal, J; Oyarzún, A; Overall, C M; Hernández, M

    2010-10-01

    To study the expression of monocyte chemotactic protein-3 (MCP-3, also known as chemokine CCL-7) in tissue from apical lesions (AL) and to associate MCP-3 expression with symptomatic or asymptomatic apical periodontitis. To determine the expression of MCP-3 in AL, biopsies obtained during tooth extraction procedures were fixed, subjected to routine processing and diagnosed as apical granuloma (AG) (n = 7) or radicular cyst (RC) (n = 5). As controls, apical periodontal ligament (PDL) specimens from healthy premolars extracted for orthodontics reasons were included (n = 7). All specimens were immunostained for MCP-3 and examined under a light microscope. In addition, homogenates from AL (n = 14) and healthy PDL samples (n = 7) were studied through immunowestern blot. Finally, periapical exudates samples were collected from root canals of teeth having diagnosis of symptomatic (n = 14) and asymptomatic apical periodontitis (n = 14) during routine endodontic treatments and analysed by immunowestern blot and densitometry.   MCP-3 was detected in AG and RC and localized mainly to inflammatory leucocytes, whereas no expression was observed in healthy PDLs. MCP-3 was also detected in periapical exudate, and its levels were significantly higher in symptomatic than in asymptomatic apical periodontitis. MCP-3 was expressed in AL and its levels associated with clinical symptoms. MCP-3 might play a role in disease pathogenesis, possibly by stimulating mononuclear chemotaxis. © 2010 International Endodontic Journal.

  4. [Radiographic and histological study of a case of apexification in a human molar].

    PubMed

    Sahli, C C

    1989-01-01

    A case of apexification in a lower right second molar is described. Radiographs demonstrate apical closure with a different morphological pattern from that of the lower left second molar. Following extraction, after 15 months, serial histologic sections show calcified tissue obturating the apical foramen, well adapted to the initial dentin and cementum walls. Inside some small areas containing connective tissue with capillaries can be observed. The histologic and radiographic observations indicate that apical closure occurs as a result of differentiation of periodontal apical cells.

  5. Dbl3 drives Cdc42 signaling at the apical margin to regulate junction position and apical differentiation

    PubMed Central

    Zihni, Ceniz; Munro, Peter M.G.; Elbediwy, Ahmed; Keep, Nicholas H.; Terry, Stephen J.; Harris, John

    2014-01-01

    Epithelial cells develop morphologically characteristic apical domains that are bordered by tight junctions, the apical–lateral border. Cdc42 and its effector complex Par6–atypical protein kinase c (aPKC) regulate multiple steps during epithelial differentiation, but the mechanisms that mediate process-specific activation of Cdc42 to drive apical morphogenesis and activate the transition from junction formation to apical differentiation are poorly understood. Using a small interfering RNA screen, we identify Dbl3 as a guanine nucleotide exchange factor that is recruited by ezrin to the apical membrane, that is enriched at a marginal zone apical to tight junctions, and that drives spatially restricted Cdc42 activation, promoting apical differentiation. Dbl3 depletion did not affect junction formation but did affect epithelial morphogenesis and brush border formation. Conversely, expression of active Dbl3 drove process-specific activation of the Par6–aPKC pathway, stimulating the transition from junction formation to apical differentiation and domain expansion, as well as the positioning of tight junctions. Thus, Dbl3 drives Cdc42 signaling at the apical margin to regulate morphogenesis, apical–lateral border positioning, and apical differentiation. PMID:24379416

  6. Efficiency of Different Endodontic Irrigation and Activation Systems in Removal of the Smear Layer: A Scanning Electron Microscopy Study.

    PubMed

    Karade, Priyatam; Chopade, Rutuja; Patil, Suvarna; Hoshing, Upendra; Rao, Madhukar; Rane, Neha; Chopade, Aditi; Kulkarni, Anish

    2017-01-01

    This in vitro study was designed to evaluate and compare different endodontic irrigation and activation systems for removal of the intracanal smear layer. Forty recently extracted, non-carious human intact single rooted premolars were selected and divided into five groups ( n =10) according to the root canal irrigation systems; syringe and needle irrigation (CTR), sonic irrigation, passive ultrasonic irrigation (PUI) and EndoVac irrigation system. All groups were prepared to #40 apical size with K-files. Each sample was subjected to final irrigation by using four different irrigation/activation systems. After splitting the samples, one half of each root was selected for examination under scanning electron microscope (SEM). The irrigation systems were compared using the Fisher's exact test with the level of significance set at 0.05. The four groups did not differ from each other in the coronal and mid-root parts of the canal. In the apical part of the canal none of the methods could completely remove all the smear layer but EndoVac system showed significantly better removal of smear layer and debris than the other methods. Within the limitations of the present study, the EndoVac system cleaned the apical part of the canal more efficiently than sonic, ultrasonic and syringe and needle irrigation.

  7. Detection of Treponema Denticola in Symptomatic Apical Periodontitis and in Symptomatic Apical Abscesses by Real-Time PCR

    PubMed Central

    Ozbek, Selcuk M.; Ozbek, Ahmet; Erdogan, Aziz S.

    2009-01-01

    Objectives: The aim of this study was to investigate the presence of Treponema denticola in symptomatic apical periodontitis and in symptomatic apical abscesses by real-time polymerase chain reaction (PCR) method. Methods: Microbial samples were collected from 60 single-rooted teeth having carious lesions and necrotic pulps. For each tooth, clinical data including patient symptoms were recorded. Teeth were categorized by diagnosis as having symptomatic apical periodontitis or symptomatic apical abscess. Aseptic microbial samples were collected using paper points from 30 infected root canals and from aspirates of 30 abscesses. DNA was extracted from the samples by using a QIAamp® DNA mini-kit and analyzed with real-time PCR. Results: T. denticola was detected in 24 of 30 cases diagnosed as symptomatic apical abscesses (80%), and 19 of 30 cases diagnosed as symptomatic apical periodontitis (63.3%). In general T. denticola was found in 43 of 60 cases (71.6%). Conclusions: Our findings suggest that T. denticola can participate in the pathogenesis of symptomatic apical abscesses. PMID:19421390

  8. Relationship between the Apical Preparation Diameter and the Apical Seal: An In Vitro Study.

    PubMed

    Laslami, Kaoutar; Dhoum, Sara; El Harchi, Amine; Benkiran, Iman

    2018-01-01

    The aim of the study is to define the relationship between the apical preparation diameter and the apical sealing ability to highlight the importance of the preservation of the diameter and the original position of the apical foramen. 50 extracted maxillary incisors were randomly allocated into three groups of 15 teeth each (n = 15) according to the apical preparation size: Group 1: finishing file F1 corresponding to size 20 reached the working length (ProTaper Universal system Dentsply®); Group 2: prepared up to size 30 corresponding to finishing file F30; Group 3: prepared up to size 50 corresponding to finishing file F5. Five teeth were assigned to positive and negative control groups. After the filling of the root canals, the teeth were isolated and immersed in a dye solution, then cut longitudinally, photographed, and the dye penetration were calculated using a computer software. Comparison of the three different apical preparation sizes showed no statistically significant differences regarding the apical microleakage. The most important value of the dye penetration was observed in the group with the largest apical diameter.

  9. An Apical MRCK-driven Morphogenetic Pathway Controls Epithelial Polarity

    PubMed Central

    Zihni, Ceniz; Vlassaks, Evi; Terry, Stephen; Carlton, Jeremy; Leung, Thomas King Chor; Olson, Michael; Pichaud, Franck; Balda, Maria Susana; Matter, Karl

    2017-01-01

    Polarized epithelia develop distinct cell surface domains, with the apical membrane acquiring characteristic morphological features such as microvilli. Cell polarization is driven by polarity determinants including the evolutionarily conserved partitioning defective (PAR) proteins that are separated into distinct cortical domains. PAR protein segregation is thought to be a consequence of asymmetric actomyosin contractions. The mechanism of activation of apically polarized actomyosin contractility is unknown. Here we show that the Cdc42 effector MRCK activates Myosin-II at the apical pole to segregate aPKC-Par6 from junctional Par3, defining the apical domain. Apically polarized MRCK-activated actomyosin contractility is reinforced by cooperation with aPKC-Par6 downregulating antagonistic RhoA-driven junctional actomyosin contractility, and drives polarization of cytosolic brush border determinants and apical morphogenesis. MRCK-activated polarized actomyosin contractility is required for apical differentiation and morphogenesis in vertebrate epithelia and Drosophila photoreceptors. Our results identify an apical origin of actomyosin-driven morphogenesis that couples cytoskeletal reorganization to PAR polarity signalling. PMID:28825699

  10. Push-out bond strengths of fiber-reinforced composite posts with various resin cements according to the root level

    PubMed Central

    Chang, Hoon-Sang; Noh, Young-Sin; Lee, Yoon; Min, Kyung-San

    2013-01-01

    PURPOSE The aim of this study was to determine whether the push-out bond strengths between the radicular dentin and fiber reinforced-composite (FRC) posts with various resin cements decreased or not, according to the coronal, middle or apical level of the root. MATERIALS AND METHODS FRC posts were cemented with one of five resin cement groups (RelyX Unicem: Uni, Contax with activator & LuxaCore-Dual: LuA, Contax & LuxaCore-Dual: Lu, Panavia F 2.0: PA, Super-Bond C&B: SB) into extracted human mandibular premolars. The roots were sliced into discs at the coronal, middle and apical levels. Push-out bond strength tests were performed with a universal testing machine at a crosshead speed of 0.5 mm/min, and the failure aspect was analyzed. RESULTS There were no significant differences (P>.05) in the bond strengths of the different resin cements at the coronal level, but there were significant differences in the bond strengths at the middle and apical levels (P<.05). Only the Uni and LuA cements did not show any significant decrease in their bond strengths at all the root levels (P>.05); all other groups had a significant decrease in bond strength at the middle or apical level (P<.05). The failure aspect was dominantly cohesive at the coronal level of all resin cements (P<.05), whereas it was dominantly adhesive at the apical level. CONCLUSION All resin cement groups showed decreases in bond strengths at the middle or apical level except LuA and Uni. PMID:24049569

  11. Novel pentapeptide, PALAL, derived from a bony fish elicits contraction of the muscle in starfish Patiria pectinifera.

    PubMed

    Go, Hye-Jin; Kim, Chan-Hee; Oh, Hye Young; Park, Nam Gyu

    2016-10-01

    A bioactive peptide mimicking peptide-signaling molecules has been isolated from the skin extract of fish Channa argus which caused contraction of the apical muscle of a starfish Patiria pectinifera, a deuterostomian invertebrate. The primary structure of the isolated pentapeptide comprises amino acid sequence of H-Pro-Ala-Leu-Ala-Leu-OH (PALAL) with a molecular mass of 483.7 Da. Pharmacological activity of PALAL, dosage ranging from 10 -9 to 10 -5 M, revealed concentration-dependent contraction of the apical muscles of P. pectinifera and Asterias amurensis. However, PALAL was not active on the intestinal smooth muscle of the goldfish Carassius auratus and has presumably other physiological roles in fish skin. Investigation of structure-activity relationship using truncated and substituted analogs of PALAL demonstrated that H-Ala-Leu-Ala-Leu-OH was necessary and should be sufficient to constrict apical muscle of P. pectinifera. Furthermore, the second alanine residue was required to display the activity, and the fifth leucine residue was responsible for its potency. Comparison with PALAL's primary structure with those of other known bioactive peptides from fish and starfish revealed that PALAL does not have any significant homology. Consequently, PALAL is a bioactive peptide that elicits a muscle contraction in starfish, and the isolation of PALAL may lead to develop other bioactive peptides sharing its similar sequence and/or activity. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

  12. Comparison of apical and coronal extrusions using reciprocating and rotary instrumentation systems.

    PubMed

    Lu, Yan; Chen, Min; Qiao, Feng; Wu, Ligeng

    2015-08-07

    The aim of this study was to compare the apical and coronal extrusions by using two reciprocating and two rotary instrumentation systems. Eighty extracted human single-rooted anterior teeth were randomly assigned to four groups. Four different root canal instrumentation systems were used according to the manufacturers instructions, including two reciprocating single-file systems, Reciproc and WaveOne, and two full-sequence rotary BLX and ProTaper instruments. Debris collected from the coronal by the instruments and apical extrusions were quantified respectively. After drying the collections, the mean weight of debris collected from apical and coronal extrusions was assessed using an electronic balance and analyzed using the Kruskal-Wallis H and Mann-Whitney U tests. Statistically significant differences in the apical extrusion were observed among the four groups. Reciproc and WaveOne instruments produced significantly less debris than BLX and ProTaper instruments (P < 0.05). All of the systems produced apical extrusion of debris. However, reciprocating single-file systems produced less apical extrusion than full-sequence rotary systems. No relationship was observed between apical and coronal extrusions.

  13. Differential signaling and regulation of apical vs. basolateral EGFR in polarized epithelial cells.

    PubMed

    Kuwada, S K; Lund, K A; Li, X F; Cliften, P; Amsler, K; Opresko, L K; Wiley, H S

    1998-12-01

    Overexpression of the epidermal growth factor receptors (EGFR) in polarized kidney epithelial cells caused them to appear in high numbers at both the basolateral and apical cell surfaces. We utilized these cells to look for differences in the regulation and signaling of apical vs. basolateral EGFR. Apical and basolateral EGFR were biologically active and mediated EGF-induced cell proliferation to similar degrees. Receptor downregulation and endocytosis were less efficient at the apical surface, resulting in prolonged EGF-induced tyrosine kinase activity at the apical cell membrane. Tyrosine phosphorylation of EGFR substrates known to mediate cell proliferation, Src-homologous and collagen protein (SHC), extracellularly regulated kinase 1 (ERK1), and ERK2 could be induced similarly by activation of apical or basolateral EGFR. Focal adhesion kinase was tyrosine phosphorylated more by basolateral than by apical EGFR; however, beta-catenin was tyrosine phosphorylated to a much greater degree following the activation of mislocalized apical EGFR. Thus EGFR regulation and EGFR-mediated phosphorylation of certain substrates differ at the apical and basolateral cell membrane domains. This suggests that EGFR mislocalization could result in abnormal signal transduction and aberrant cell behavior.

  14. Prevalence of ciliated epithelium in apical periodontitis lesions.

    PubMed

    Ricucci, Domenico; Loghin, Simona; Siqueira, José F; Abdelsayed, Rafik A

    2014-04-01

    This article reports on the morphologic features and the frequency of ciliated epithelium in apical cysts and discusses its origin. The study material consisted of 167 human apical periodontitis lesions obtained consecutively from patients presenting for treatment during a period of 12 years in a dental practice operated by one of the authors. All of the lesions were obtained still attached to the root apices of teeth with untreated (93 lesions) or treated canals (74 lesions). The former were obtained by extraction and the latter by extraction or apical surgery. Specimens were processed for histopathologic and histobacteriologic analyses. Lesions were classified, and the type of epithelium, if present, was recorded. Of the lesions analyzed, 49 (29%) were diagnosed as cysts. Of these, 26 (53%) were found in untreated teeth, and 23 (47%) related to root canal-treated teeth. Ciliated columnar epithelium was observed partially or completely lining the cyst wall in 4 cysts, and all of them occurred in untreated maxillary molars. Three of these lesions were categorized as pocket cysts, and the other was a true cyst. Ciliated columnar epithelium-lined cysts corresponded to approximately 2% of the apical periodontitis lesions and 8% of the cysts of endodontic origin in the population studied. This epithelium is highly likely to have a sinus origin in the majority of cases. However, the possibility of prosoplasia or upgraded differentiation into ciliated epithelium from the typical cystic lining squamous epithelium may also be considered. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  15. Apical extrusion of debris in four different endodontic instrumentation systems: A meta-analysis.

    PubMed

    Western, J Sylvia; Dicksit, Daniel Devaprakash

    2017-01-01

    All endodontic instrumentation systems tested so far, promote apical extrusion of debris, which is one of the main causes of postoperative pain, flare ups, and delayed healing. Of this meta-analysis was to collect and analyze in vitro studies quantifying apically extruded debris while using Hand ProTaper (manual), ProTaper Universal (rotary), Wave One (reciprocating), and self-adjusting file (SAF; vibratory) endodontic instrumentation systems and to determine methods which produced lesser extrusion of debris apically. An extensive electronic database search was done in PubMed, Scopus, Cochrane, LILACS, and Google Scholar from inception until February 2016 using the key terms "Apical Debris Extrusion, extruded material, and manual/rotary/reciprocating/SAF systems." A systematic search strategy was followed to extract 12 potential articles from a total of 1352 articles. The overall effect size was calculated from the raw mean difference of weight of apically extruded debris. Statistically significant difference was seen in the following comparisons: SAF < Wave One, SAF < Rotary ProTaper. Apical extrusion of debris was invariably present in all the instrumentation systems analyzed. SAF system seemed to be periapical tissue friendly as it caused reduced apical extrusion compared to Rotary ProTaper and Wave One.

  16. Incidence of apical crack formation and propagation during removal of root canal filling materials with different engine driven nickel-titanium instruments.

    PubMed

    Özyürek, Taha; Tek, Vildan; Yılmaz, Koray; Uslu, Gülşah

    2017-11-01

    To determine the incidence of crack formation and propagation in apical root dentin after retreatment procedures performed using ProTaper Universal Retreatment (PTR), Mtwo-R, ProTaper Next (PTN), and Twisted File Adaptive (TFA) systems. The study consisted of 120 extracted mandibular premolars. One millimeter from the apex of each tooth was ground perpendicular to the long axis of the tooth, and the apical surface was polished. Twenty teeth served as the negative control group. One hundred teeth were prepared, obturated, and then divided into 5 retreatment groups. The retreatment procedures were performed using the following files: PTR, Mtwo-R, PTN, TFA, and hand files. After filling material removal, apical enlargement was done using apical size 0.50 mm ProTaper Universal (PTU), Mtwo, PTN, TFA, and hand files. Digital images of the apical root surfaces were recorded before preparation, after preparation, after obturation, after filling removal, and after apical enlargement using a stereomicroscope. The images were then inspected for the presence of new apical cracks and crack propagation. Data were analyzed with χ 2 tests using SPSS 21.0 software. New cracks and crack propagation occurred in all the experimental groups during the retreatment process. Nickel-titanium rotary file systems caused significantly more apical crack formation and propagation than the hand files. The PTU system caused significantly more apical cracks than the other groups after the apical enlargement stage. This study showed that retreatment procedures and apical enlargement after the use of retreatment files can cause crack formation and propagation in apical dentin.

  17. [Cytocompatibility of Co-Cr ceramic alloys after recasting].

    PubMed

    Hu, Yu-Feng; Jin, Wen-Zhong

    2017-06-01

    To study the correlation between apical foramen area and accuracy of PropexII electronic apex locator under destroyed apical constriction. Forty extracted teeth with single straight root canal were ground down 1 mm in the root tip and placed in 2% liquid agar gel injected into Castro model. The length of root canal was measured by PropexII electronic apex locator. The difference (L) between the electronic length (LP) and actual length was calculated. Imaging of apical foramen was recorded under microscope and apical foramen area (S) was measured by image processing software Photoshop CS. SPSS 22.0 software package was used to analyze the linear correlation and regression. With ±0.5 mm as the allowable range, all value of L was positive. The precise rate of PropexII was 52.5% when apical constriction was destroyed. There was a linear relationship between S and L (S=0.04+0.11×L,R=0.903). The accuracy decreases when apical constriction is destroyed. The accuracy is worse when the apical foramen area is larger.

  18. Pollen-tube tip growth requires a balance of lateral propagation and global inhibition of Rho-family GTPase activity

    PubMed Central

    Hwang, Jae-Ung; Wu, Guang; Yan, An; Lee, Yong-Jik; Grierson, Claire S.; Yang, Zhenbiao

    2010-01-01

    Rapid tip growth allows for efficient development of highly elongated cells (e.g. neuronal axons, fungal hyphae and pollen tubes) and requires an elaborate spatiotemporal regulation of the growing region. Here, we use the pollen tube as a model to investigate the mechanism regulating the growing region. ROPs (Rho-related GTPases from plants) are essential for pollen tip growth and display oscillatory activity changes in the apical plasma membrane (PM). By manipulating the ROP activity level, we showed that the PM distribution of ROP activity as an apical cap determines the tip growth region and that efficient tip growth requires an optimum level of the apical ROP1 activity. Excessive ROP activation induced the enlargement of the tip growth region, causing growth depolarization and reduced tube elongation. Time-lapse analysis suggests that the apical ROP1 cap is generated by lateral propagation of a localized ROP activity. Subcellular localization and gain- and loss-of-function analyses suggest that RhoGDI- and RhoGAP-mediated global inhibition limits the lateral propagation of apical ROP1 activity. We propose that the balance between the lateral propagation and the global inhibition maintains an optimal apical ROP1 cap and generates the apical ROP1 activity oscillation required for efficient pollen-tube elongation. PMID:20053639

  19. Cellular Profile and Expression of Immunologic Markers in Chronic Apical Periodontitis from HIV-infected Patients Undergoing Highly Active Antiretroviral Therapy.

    PubMed

    Gama, Túlio Gustavo Veiga; Pires, Fabio Ramoa; Armada, Luciana; Gonçalves, Lucio Souza

    2016-06-01

    This study tested the hypothesis that the inflammatory cell profile (CD3-, CD4-, CD8-, CD20-, and CD68-positive cells) and the expression of immunologic markers (tumor necrosis factor α, interferon-γ, interleukin-6, and interleukin-18) in chronic apical periodontitis are the same between non-HIV-infected patients and HIV-infected patients undergoing highly active antiretroviral therapy (HAART). Thirty-four surgically excised chronic apical periodontitis lesions were sampled from 34 patients (17 HIV-infected and 17 non-HIV-infected). The lesions were extracted from teeth with no previous endodontic treatment. All HIV-infected patients were undergoing HAART. The specimens were submitted to histopathologic and immunohistochemical analyses by using an optical microscope. Immunoexpression was graded into 2 levels, focal to weak and moderate to strong. The χ(2), Fisher exact, and Mann-Whitney tests were used to analyze all significant differences between groups. Periapical cysts represented 70.6% and 52.9% of the lesions in the HIV-infected and non-HIV-infected groups, respectively; however, no statistically significant difference was observed (P = .481). There were no statistically significant differences between groups for the inflammatory cell profile and for any of the immunologic markers (P > .05). There are no statistically significant differences of the cellular profile and expression of immunologic markers in chronic apical periodontitis between non-HIV-infected patients and HIV-infected patients undergoing HAART. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  20. MRCK-1 drives apical constriction in C. elegans by linking developmental patterning to force generation

    PubMed Central

    Marston, Daniel J.; Higgins, Christopher D.; Peters, Kimberly A.; Cupp, Timothy D.; Dickinson, Daniel J.; Pani, Ariel M.; Moore, Regan P.; Cox, Amanda H.; Kiehart, Daniel P.; Goldstein, Bob

    2016-01-01

    Summary Apical constriction is a change in cell shape that drives key morphogenetic events including gastrulation and neural tube formation. Apical force-producing actomyosin networks drive apical constriction by contracting while connected to cell-cell junctions. The mechanisms by which developmental patterning regulates these actomyosin networks and associated junctions with spatial precision are not fully understood. Here, we identify a myosin light chain kinase MRCK-1 as a key regulator of C. elegans gastrulation that integrates spatial and developmental patterning information. We show that MRCK-1 is required for activation of contractile actomyosin dynamics and elevated cortical tension in the apical cell cortex of endodermal precursor cells. MRCK-1 is apically localized by active Cdc42 at the external, cell-cell contact-free surfaces of apically constricting cells, downstream of cell fate determination mechanisms. We establish that the junctional components α-catenin, β-catenin, and cadherin become highly enriched at the apical junctions of apically-constricting cells, and that MRCK-1 and myosin activity are required in vivo for this enrichment. Taken together, our results define mechanisms that position a myosin activator to a specific cell surface where it both locally increases cortical tension and locally enriches junctional components to facilitate apical constriction. These results reveal crucial links that can tie spatial information to local force generation to drive morphogenesis. PMID:27451898

  1. Apical debris extrusion associated with oval shaped canals: a comparative study of WaveOne vs Self-Adjusting File.

    PubMed

    Farmakis, Eleftherios Terry R; Sotiropoulos, G G; Abràmovitz, I; Solomonov, M

    2016-11-01

    The aim was to evaluate ex vivo apical debris extrusion associated with WaveOne and Self-Adjusting File instruments when used in oval canals. Twenty-four extracted human mandibular premolars with oval-shaped canals were assigned in two equal groups. Following coronal cavity preparation, a glide path was created. Group A was subjected to canal preparation using a WaveOne primary file, which was used along with syringe and needle irrigation and 10 mL of 2.4 % NaOCl solution, followed by flushing with 10 mL of 17 % EDTA solution, activation with EndoActivator for 1 min and final flushing with 10 mL of 2.4 % NaOCl solution, and activation for 30 s. Group B, the SAF system was used with continuous simultaneous irrigation, provided by the system's pump. The irrigant was supplied at 5 mL/min, alternating every minute between 2.4 % NaOCl solution and 17 % EDTA solution, over a total of 4 min followed by final flushing with 10 mL of 2.4 % NaOCl solution. Extruded apical debris from each root canal was collected into a preweighed glass vial and dried. The mean weight of the debris from each group was assessed and analyzed statistically. Both systems resulted in apical debris extrusion. The WaveOne system was associated with a statistically significant greater mean mass of apically extruded debris (2.18 ± 0.44 mg) than the SAF system (0.49 ± 0.33 mg, permutation-based Wilcoxon test, p < 0.001). Both WaveOne and the SAF systems were associated with apical debris extrusion. The amount of debris extruded by the WaveOne system was 4.4 times greater than that extruded by the SAF system. The results of the present ex vivo comparative study cannot be directly applied to the clinical situation. Difference between both groups remains completely unclear; maybe the amount of extrusion is harmless in both groups or similarly deleterious for the periradicular tissues in both groups or may be dose-related to the amount of the extruded material.

  2. Abscisic Acid and the Photoperiodic Induction of Dormancy in Salix viminalis L.

    PubMed

    Alvim, R; Saunders, P F; Barros, R S

    1979-04-01

    A series of growth room experiments was carried out aiming to establish the role of abscisic acid on dormancy of Salix viminalis L. The inhibitor content and abscisic acid levels of extracts from roots, sap, leaves, and apical tissues of willow were measured using biological assay and gas-liquid chromatography.No evidence was obtained that photoperiodically mediated dormancy is associated with changes in abscisic acid levels or beta-inhibitor activity.

  3. In vitro evaluation of apical extrusion of bacteria following use of new rotary instrumentation system.

    PubMed

    Mohammadi, Zahed

    2009-04-01

    The aim of this study was to evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using two engine-driven nickel-titanium instruments (Flex Master and V-Taper). Seventy extracted maxillary central incisor teeth were used. After preparing access cavities, root canals were contaminated with a suspension of Enterococcus faecalis, then dried. The contaminated roots were divided into two experimental groups of 30 teeth each and one control group of 10 teeth. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 hours. Colonies of bacteria were counted, and the results were given as number of colony-forming units. The data obtained were analyzed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests, with alpha = 0.05 as the level for statistical significance. Results showed that there was no significant difference as to the number of extruded bacteria between the two engine-driven systems (P > 0.05). Both engine-driven nickel-titanium systems extruded bacteria through the apical foramen.

  4. Distribution and Function of HCN Channels in the Apical Dendritic Tuft of Neocortical Pyramidal Neurons

    PubMed Central

    Harnett, Mark T.; Magee, Jeffrey C.

    2015-01-01

    The apical tuft is the most remote area of the dendritic tree of neocortical pyramidal neurons. Despite its distal location, the apical dendritic tuft of layer 5 pyramidal neurons receives substantial excitatory synaptic drive and actively processes corticocortical input during behavior. The properties of the voltage-activated ion channels that regulate synaptic integration in tuft dendrites have, however, not been thoroughly investigated. Here, we use electrophysiological and optical approaches to examine the subcellular distribution and function of hyperpolarization-activated cyclic nucleotide-gated nonselective cation (HCN) channels in rat layer 5B pyramidal neurons. Outside-out patch recordings demonstrated that the amplitude and properties of ensemble HCN channel activity were uniform in patches excised from distal apical dendritic trunk and tuft sites. Simultaneous apical dendritic tuft and trunk whole-cell current-clamp recordings revealed that the pharmacological blockade of HCN channels decreased voltage compartmentalization and enhanced the generation and spread of apical dendritic tuft and trunk regenerative activity. Furthermore, multisite two-photon glutamate uncaging demonstrated that HCN channels control the amplitude and duration of synaptically evoked regenerative activity in the distal apical dendritic tuft. In contrast, at proximal apical dendritic trunk and somatic recording sites, the blockade of HCN channels decreased excitability. Dynamic-clamp experiments revealed that these compartment-specific actions of HCN channels were heavily influenced by the local and distributed impact of the high density of HCN channels in the distal apical dendritic arbor. The properties and subcellular distribution pattern of HCN channels are therefore tuned to regulate the interaction between integration compartments in layer 5B pyramidal neurons. PMID:25609619

  5. Comparative evaluation of the shaping ability of two different nickel-titanium rotary files in curved root canals of extracted human molar teeth.

    PubMed

    Ferrara, Giulia; Taschieri, Silvio; Corbella, Stefano; Ceci, Caterina; Del Fabbro, Massimo; Machtou, Pierre

    2017-02-01

    The aim of the present study was to compare the shaping ability of two different nickel-titanium rotary files in the curved root canals of extracted human molar teeth. Thirty root canals of 17 extracted human molars teeth were randomly assigned to two experimental groups (n = 15): ProTaper Next and ProTaper Universal (PTU), on the basis of the rotary files system used. The final size of all apical foramina was 0.25 mm in diameter. Standardized digital radiographs were taken before and after instrumentation in both clinical and proximal views, with a size 10 K-file inserted into the canal for the determination of the angle of curvature and apical transportation. Preparation time and fractured or deformed instruments were also recorded. The unpaired Student's t-test was used to compare results. There was no significant difference between the two instruments with respect to canal straightening and apical transportation before and after instrumentation (P > 0.05). The use of both instruments resulted in a significant reduction in the angle of curvature after instrumentation (P < 0.05). Instrumentation time was significantly greater for PTU (P < 0.05). The ProTaper Universal and ProTaper Next systems performed similarly with regard to the straightening of curved root canals and apical transportation. ProTaper Next was significantly faster than ProTaper Universal. © 2015 Wiley Publishing Asia Pty Ltd.

  6. A comparison of apical transportation between FlexMaster and Twisted Files rotary instruments.

    PubMed

    Duran-Sindreu, Fernando; García, Marc; Olivieri, Juan Gonzalo; Mercadé, Montse; Morelló, Sergio; Roig, Miguel

    2012-07-01

    The aim of this study was to evaluate apical transportation in root canals after the use of Twisted Files (TF; SybronEndo, Orange, CA) and FlexMaster (VDW, Munich, Germany) #40/04 rotary files. A double-digital radiographic technique was used to compare apical transportation between the TF and FlexMaster systems. Each rotary system was used to instrument mesial canals from 80 extracted mandibular molars. The central axes of the file imaged before instrumentation (#15 K-file) and the master apical rotary file (#40/04) were superimposed digitally. AutoCAD 2008 (Autodesk Inc, San Rafael, CA) was used to measure apical transportation at 0.5 mm from the working length (WL). The data were analyzed using the Student's t test, and significance was set at P < .05. The mean amount of apical transportation at 0.5 mm from the WL was 0.17 ± 0.09 mm for the FlexMaster group and 0.19 ± 0.12 mm for the TF group. No statistically significant differences in apical transportation were found between the 2 groups. Under the conditions of the study, no statistically significant differences in apical transportation were observed between FlexMaster and TF rotary files. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  7. Apical extrusion of debris in four different endodontic instrumentation systems: A meta-analysis

    PubMed Central

    Western, J. Sylvia; Dicksit, Daniel Devaprakash

    2017-01-01

    Background: All endodontic instrumentation systems tested so far, promote apical extrusion of debris, which is one of the main causes of postoperative pain, flare ups, and delayed healing. Objectives: Of this meta-analysis was to collect and analyze in vitro studies quantifying apically extruded debris while using Hand ProTaper (manual), ProTaper Universal (rotary), Wave One (reciprocating), and self-adjusting file (SAF; vibratory) endodontic instrumentation systems and to determine methods which produced lesser extrusion of debris apically. Methodology: An extensive electronic database search was done in PubMed, Scopus, Cochrane, LILACS, and Google Scholar from inception until February 2016 using the key terms “Apical Debris Extrusion, extruded material, and manual/rotary/reciprocating/SAF systems.” A systematic search strategy was followed to extract 12 potential articles from a total of 1352 articles. The overall effect size was calculated from the raw mean difference of weight of apically extruded debris. Results: Statistically significant difference was seen in the following comparisons: SAF < Wave One, SAF < Rotary ProTaper. Conclusions: Apical extrusion of debris was invariably present in all the instrumentation systems analyzed. SAF system seemed to be periapical tissue friendly as it caused reduced apical extrusion compared to Rotary ProTaper and Wave One. PMID:28761250

  8. Mitogen-activated protein kinases participate in determination of apical-basal symmetry in Pisum sativum.

    PubMed

    Winnicki, Konrad; Polit, Justyna Teresa; Żabka, Aneta; Maszewski, Janusz

    2017-03-01

    Mitogen-activated protein kinases (MAPKs) are implicated in various processes in plants. Apart from response to biotic and abiotic stresses they are involved in regulation of embryo development. Although MAPKs were found to be indispensable during embryo development it has never been established at which stages of embryogenesis and in which regions of a plant embryo activated MAPKs can be observed. Here, we show that apical and basal regions display activation of the same types of MAPKs and the only difference concerns the level of their phosphorylation and cellular localization. Dually-phosphorylated MAPKs were found in nuclei of the apical region of an embryo both at the early and late cotyledonary stage while no immunofluorescence signals were detected in nuclei of the basal region. However, in this case phosphorylated MAPKs were immunodetected in cytoplasm in the apical domain of cortex cells, indicating their role in auxin transport from the basal to apical region of an embryo. Furthermore, obtained data indicate that nuclear localization of activated MAPKs may result from epigenetic modifications and polar auxin transport. The presented data and previous studies lead to the conclusion that activated MAPKs and their cellular localization define apical and basal regions during formation of an apical-basal axis. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Apical targeting of the formin Diaphanous in Drosophila tubular epithelia

    PubMed Central

    Rousso, Tal; Shewan, Annette M; Mostov, Keith E; Schejter, Eyal D; Shilo, Ben-Zion

    2013-01-01

    Apical secretion from epithelial tubes of the Drosophila embryo is mediated by apical F-actin cables generated by the formin-family protein Diaphanous (Dia). Apical localization and activity of Dia are at the core of restricting F-actin formation to the correct membrane domain. Here we identify the mechanisms that target Dia to the apical surface. PI(4,5)P2 levels at the apical membrane regulate Dia localization in both the MDCK cyst model and in Drosophila tubular epithelia. An N-terminal basic domain of Dia is crucial for apical localization, implying direct binding to PI(4,5)P2. Dia apical targeting also depends on binding to Rho1, which is critical for activation-induced conformational change, as well as physically anchoring Dia to the apical membrane. We demonstrate that binding to Rho1 facilitates interaction with PI(4,5)P2 at the plane of the membrane. Together these cues ensure efficient and distinct restriction of Dia to the apical membrane. DOI: http://dx.doi.org/10.7554/eLife.00666.001 PMID:23853710

  10. Histobacteriologic Conditions of the Apical Root Canal System and Periapical Tissues in Teeth Associated with Sinus Tracts.

    PubMed

    Ricucci, Domenico; Loghin, Simona; Gonçalves, Lucio S; Rôças, Isabela N; Siqueira, José F

    2018-03-01

    This histobacteriologic study described the pattern of intraradicular and extraradicular infections in teeth with sinus tracts and chronic apical abscesses. The material comprised biopsy specimens from 24 (8 untreated and 16 treated) roots of teeth associated with apical periodontitis and a sinus tract. Specimens were obtained by periradicular surgery or extraction and were processed for histobacteriologic and histopathologic methods. Bacteria were found in the apical root canal system of all specimens, in the main root canal (22 teeth) and within ramifications (17 teeth). Four cases showed no extraradicular infection. Extraradicular bacteria occurred as a biofilm attached to the outer root surface in 17 teeth (5 untreated and 12 treated teeth), as actinomycotic colonies in 2 lesions, and as planktonic cells in 2 lesions. Extraradicular calculus formation (mineralized biofilm) was evident in 10 teeth. Teeth with chronic apical abscesses and sinus tracts showed a very complex infectious pattern in the apical root canal system and periapical lesion, with a predominance of biofilms. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  11. Distribution and function of HCN channels in the apical dendritic tuft of neocortical pyramidal neurons.

    PubMed

    Harnett, Mark T; Magee, Jeffrey C; Williams, Stephen R

    2015-01-21

    The apical tuft is the most remote area of the dendritic tree of neocortical pyramidal neurons. Despite its distal location, the apical dendritic tuft of layer 5 pyramidal neurons receives substantial excitatory synaptic drive and actively processes corticocortical input during behavior. The properties of the voltage-activated ion channels that regulate synaptic integration in tuft dendrites have, however, not been thoroughly investigated. Here, we use electrophysiological and optical approaches to examine the subcellular distribution and function of hyperpolarization-activated cyclic nucleotide-gated nonselective cation (HCN) channels in rat layer 5B pyramidal neurons. Outside-out patch recordings demonstrated that the amplitude and properties of ensemble HCN channel activity were uniform in patches excised from distal apical dendritic trunk and tuft sites. Simultaneous apical dendritic tuft and trunk whole-cell current-clamp recordings revealed that the pharmacological blockade of HCN channels decreased voltage compartmentalization and enhanced the generation and spread of apical dendritic tuft and trunk regenerative activity. Furthermore, multisite two-photon glutamate uncaging demonstrated that HCN channels control the amplitude and duration of synaptically evoked regenerative activity in the distal apical dendritic tuft. In contrast, at proximal apical dendritic trunk and somatic recording sites, the blockade of HCN channels decreased excitability. Dynamic-clamp experiments revealed that these compartment-specific actions of HCN channels were heavily influenced by the local and distributed impact of the high density of HCN channels in the distal apical dendritic arbor. The properties and subcellular distribution pattern of HCN channels are therefore tuned to regulate the interaction between integration compartments in layer 5B pyramidal neurons. Copyright © 2015 the authors 0270-6474/15/351024-14$15.00/0.

  12. Localization and signaling patterns of vascular endothelial growth factors and receptors in human periapical lesions.

    PubMed

    Virtej, Anca; Løes, Sigbjørn S; Berggreen, Ellen; Bletsa, Athanasia

    2013-05-01

    Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are key players in vasculogenesis and are also involved in pathologic conditions with bone destruction. Vasculogenesis is critical for disease progression, and bone resorption is a hallmark of apical periodontitis. However, the localization of VEGFs and VEGFRs and their gene signaling pathways in human apical periodontitis have not been thoroughly investigated. The aim of this study was to localize VEGFs and VEGFRs and analyze their gene expression as well as signaling pathways in human periapical lesions. Tissue was collected after endodontic surgery from patients diagnosed with chronic apical periodontitis. Periodontal ligament samples from extracted healthy wisdom teeth was also collected and used as control tissue. In lesion cryosections, VEGFs/VEGFRs were identified by immunohistochemistry/double immunofluorescence by using specific antibodies. A human VEGF signaling polymerase chain reaction array system was used for gene expression analysis comparing lesions with periodontal ligament samples. The histologic evaluation revealed heterogeneous morphology of the periapical lesions with various degrees of inflammatory infiltrates. In the lesions, all investigated factors and receptors were identified in blood vessels and various immune cells. No lymphatic vessels were detected. Gene expression analysis revealed up-regulation of VEGF-A and VEGFR-3, although not significant. Phosphatidylinositol-3-kinases, protein kinase C, mitogen-activated protein kinases, and phospholipases, all known to be involved in VEGF-mediated angiogenic activity, were significantly up-regulated. The cellular and vascular expressions of VEGFs and VEGFRs in chronic apical periodontitis, along with significant alterations of genes mediating VEGF-induced angiogenic responses, suggest ongoing vascular remodeling in established chronic periapical lesions. Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  13. Quantitative evaluation of apical extrusion of debris and irrigants using four rotary instrumentation systems: an in vitro study.

    PubMed

    Nagaveni, S Aspalli; Balakoti, K Reddy; Smita, Karan; Ratnakar, P; Satish, S V; Aravind, T

    2013-11-01

    The apical extrusion of infected debris may have the potential to disrupt the balance between microbial aggression and host defense, resulting in incidents of acute inflammation. During preparation, irrigants and debris, such as bacteria, dentin filings and necrotic tissue may be extruded into the periradicular region leading to periapical inflammation and postoperative flare ups. Using an instrumentation technique that minimizes apical extrusion would be beneficial to both the practitioner and patient. The purpose of the study was to evaluate the weight of debris and volume of irrigant extruded apically from extracted teeth in vitro after endodontic instrumentation using four different rotary root canal instrumentation systems. Four groups of each 20 extracted mandibular premolars were instrumented using one of the four systems: ProTaper Universal (Dentsply Maillefer, Ballaigues, Switzerland)), Hero-shaper (MicroMega, Besancon, France), RaCe (FKG Dentaire, La-Chaux-de-Fonds, Switzerland) and K3 (SybronEndo, West Collins, CA). Debris and irrigant extruded from the apical foramen during instrumentation were collected in preweighed test tubes. Volume of irrigant extruded was noted. The containers were stored in incubator at 70° for two days to evaporate the moisture. Weight of dry debris was noted. Data was analyzed using Kruskall-Wallis and Mann-Whitney U test at a significance of 0.001. The results indicated that all of the instrumentation systems tested caused measurable apical extrusion of debris and irrigants. Higher extrusion was observed with Protaper system which was statistically significant with Hero-Shaper, RaCe and K3 systems. There were no statistical differences between Hero-shaper, K3 and RaCe systems (p < 0.05). All instrumentation techniques apically extruded debris and irrigant. However, Hero-shaper, K3 and RaCe systems produced less extruded debris and irrigant than the Protaper system.

  14. Abscisic Acid and the Photoperiodic Induction of Dormancy in Salix viminalis L 1

    PubMed Central

    Alvim, Ronald; Saunders, Peter F.; Barros, Raimundo S.

    1979-01-01

    A series of growth room experiments was carried out aiming to establish the role of abscisic acid on dormancy of Salix viminalis L. The inhibitor content and abscisic acid levels of extracts from roots, sap, leaves, and apical tissues of willow were measured using biological assay and gas-liquid chromatography. No evidence was obtained that photoperiodically mediated dormancy is associated with changes in abscisic acid levels or β-inhibitor activity. PMID:16660810

  15. Neisseria gonorrhoeae breaches the apical junction of polarized epithelial cells for transmigration by activating EGFR

    PubMed Central

    Edwards, Vonetta L.; Wang, Liang-Chun; Dawson, Valerie; Stein, Daniel C.; Song, Wenxia

    2017-01-01

    Summary Neisseria gonorrhoeae initiates infection at the apical surface of columnar endocervical epithelial cells in the female reproductive tract. These cells provide a physical barrier against pathogens by forming continuous apical junctional complexes between neighbouring cells. This study examines the interaction of gonococci (GC) with polarized epithelial cells. We show that viable GC preferentially localize at the apical side of the cell–cell junction in polarized endometrial and colonic epithelial cells, HEC-1-B and T84. In GC-infected cells, continuous apical junctional complexes are disrupted, and the junction-associated protein β-catenin is redistributed from the apical junction to the cytoplasm and to GC adherent sites; however, overall cellular levels remain unchanged. This redistribution of junctional proteins is associated with a decrease in the ‘fence’ function of the apical junction but not its ‘gate’ function. Disruption of the apical junction by removing calcium increases GC transmigration across the epithelial monolayer. GC inoculation induces the phosphorylation of both epidermal growth factor receptor (EGFR) and β-catenin, while inhibition of EGFR kinase activity significantly reduces both GC-induced β-catenin redistribution and GC transmigration. Therefore, the gonococcus is capable of weakening the apical junction and polarity of epithelial cells by activating EGFR, which facilitates GC transmigration across the epithelium. PMID:23279089

  16. Apical Na(+)-D-glucose cotransporter 1 (SGLT1) activity and protein abundance are expressed along the jejunal crypt-villus axis in the neonatal pig

    USDA-ARS?s Scientific Manuscript database

    Gut apical Na(+)-glucose cotransporter 1 (SGLT1) activity is high at the birth and during suckling, thus contributing substantially to neonatal glucose homeostasis. We hypothesize that neonates possess high SGLT1 maximal activity by expressing apical SGLT1 protein along the intestinal crypt-villus a...

  17. Microbiome in the Apical Root Canal System of Teeth with Post-Treatment Apical Periodontitis

    PubMed Central

    Siqueira, José F.; Antunes, Henrique S.; Rôças, Isabela N.; Rachid, Caio T. C. C.

    2016-01-01

    Introduction Bacteria present in the apical root canal system are directly involved with the pathogenesis of post-treatment apical periodontitis. This study used a next-generation sequencing approach to identify the bacterial taxa occurring in cryopulverized apical root samples from root canal-treated teeth with post-treatment disease. Methods Apical root specimens obtained during periradicular surgery of ten adequately treated teeth with persistent apical periodontitis were cryogenically ground. DNA was extracted from the powder and the microbiome was characterized on the basis of the V4 hypervariable region of the 16S rRNA gene by using paired-end sequencing on Illumina MiSeq device. Results All samples were positive for the presence of bacterial DNA. Bacterial taxa were mapped to 11 phyla and 103 genera composed by 538 distinct operational taxonomic units (OTUs) at 3% of dissimilarity. Over 85% of the sequences belonged to 4 phyla: Proteobacteria, Firmicutes, Fusobacteria and Actinobacteria. In general, these 4 phyla accounted for approximately 80% of the distinct OTUs found in the apical root samples. Proteobacteria was the most abundant phylum in 6/10 samples. Fourteen genera had representatives identified in all cases. Overall, the genera Fusobacterium and Pseudomonas were the most dominant. Enterococcus was found in 4 cases, always in relatively low abundance. Conclusions This study showed a highly complex bacterial community in the apical root canal system of adequately treated teeth with persistent apical periodontitis. This suggests that this disease is characterized by multispecies bacterial communities and has a heterogeneous etiology, because the community composition largely varied from case to case. PMID:27689802

  18. Microbiome in the Apical Root Canal System of Teeth with Post-Treatment Apical Periodontitis.

    PubMed

    Siqueira, José F; Antunes, Henrique S; Rôças, Isabela N; Rachid, Caio T C C; Alves, Flávio R F

    Bacteria present in the apical root canal system are directly involved with the pathogenesis of post-treatment apical periodontitis. This study used a next-generation sequencing approach to identify the bacterial taxa occurring in cryopulverized apical root samples from root canal-treated teeth with post-treatment disease. Apical root specimens obtained during periradicular surgery of ten adequately treated teeth with persistent apical periodontitis were cryogenically ground. DNA was extracted from the powder and the microbiome was characterized on the basis of the V4 hypervariable region of the 16S rRNA gene by using paired-end sequencing on Illumina MiSeq device. All samples were positive for the presence of bacterial DNA. Bacterial taxa were mapped to 11 phyla and 103 genera composed by 538 distinct operational taxonomic units (OTUs) at 3% of dissimilarity. Over 85% of the sequences belonged to 4 phyla: Proteobacteria, Firmicutes, Fusobacteria and Actinobacteria. In general, these 4 phyla accounted for approximately 80% of the distinct OTUs found in the apical root samples. Proteobacteria was the most abundant phylum in 6/10 samples. Fourteen genera had representatives identified in all cases. Overall, the genera Fusobacterium and Pseudomonas were the most dominant. Enterococcus was found in 4 cases, always in relatively low abundance. This study showed a highly complex bacterial community in the apical root canal system of adequately treated teeth with persistent apical periodontitis. This suggests that this disease is characterized by multispecies bacterial communities and has a heterogeneous etiology, because the community composition largely varied from case to case.

  19. Survival and genetic stability of Dendranthema grandiflora Tzvelev shoot apices after cryopreservation by vitrification and encapsulation-dehydration.

    PubMed

    Martín, Carmen; González-Benito, M Elena

    2005-12-01

    The aim of this study was to compare the genetic stability of chrysanthemum (cv. Pasodoble) apices cryopreserved using two different methods: encapsulation-dehydration and vitrification. The assessment of the genetic stability was developed using RAPDs markers. Assessment of stability was evaluated in pot-cultivated mother plants (from which buds were excised for micropropagation), in shoots (leave tissue) from which apices were extracted for cryopreservation, and in shoots regenerated from cryopreserved apices 30 days after recovery and after further 3 months in culture. Throughout the process the origin of the apices (in vitro-shoot from which they were excised) was recorded. Twenty one regenerants cryopreserved by vitrification and 25 by encapsulation-dehydration were assessed. Only one cryopreserved regenerant from the encapsulation-dehydration method showed a different band pattern. These results support the necessity of monitoring the genetic stability of the regenerants obtained after cryopreservation, as this is a very useful technique for the conservation of plant genetic resources.

  20. Reciproc versus Twisted file for root canal filling removal: assessment of apically extruded debris.

    PubMed

    Altunbas, Demet; Kutuk, Betul; Toyoglu, Mustafa; Kutlu, Gizem; Kustarci, Alper; Er, Kursat

    2016-01-01

    The aim of this study was to evaluate the amount of apically extruded debris during endodontic retreatment with different file systems. Sixty extracted human mandibular premolar teeth were used in this study. Root canals of the teeth were instrumented and filled before being randomly assigned to three groups. Guttapercha was removed using the Reciproc system, the Twisted File system (TF), and Hedström-files (H-file). Apically extruded debris was collected and dried in pre-weighed Eppendorf tubes. The amount of extruded debris was assessed with an electronic balance. Data were statistically analyzed using one-way ANOVA, Kruskal-Wallis, and Mann-Whitney U tests. The Reciproc and TF systems extruded significantly less debris than the H-file (p<0.05). However, no significant difference was found between the Reciproc and TF systems. All tested file systems caused apical extrusion of debris. Both the rotary file (TF) and the reciprocating single-file (Reciproc) systems were associated with less apical extrusion compared with the H-file.

  1. Herpesviruses in asymptomatic apical periodontitis lesions: an immunohistochemical approach.

    PubMed

    Saboia-Dantas, C J; Coutrin de Toledo, L F; Sampaio-Filho, H R; Siqueira, J F

    2007-10-01

    Human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) have been recently detected in samples from apical periodontitis lesions by means of molecular biology techniques and a role in the pathogenesis of this disease has been suggested. The present study was designed to survey asymptomatic primary apical periodontitis lesions for the presence of HCMV- and/or EBV-infected cells by means of immunohistochemistry. Apical periodontitis lesions were obtained from 35 patients [26 human immunodeficiency virus (HIV) -seronegative patients and nine HIV-seropositive patients] after tooth extraction and subjected to immunohistochemical analysis using monoclonal antibodies specific for HCMV and EBV. Fifteen of the 35 apical periodontitis lesions were positive for the target herpesviruses. Overall, EBV was found in 31% of the samples and HCMV in 23%, with 14% of the lesions showing EBV and HCMV dual infection. No association was found between HCMV or EBV with any particular histopathological type of apical periodontitis (P > 0.05). HCMV was significantly more frequent in apical periodontitis lesions from HIV-positive patients (67%) than in lesions from HIV-negative patients (8%) (P = 0.001). EBV was detected in 44% of lesions from HIV-positive patients and in 27% of lesions from HIV-negative patients, but this difference was not significant (P = 0.91). Our results showed that cells infected by HCMV and EBV can be found in apical periodontitis lesions, with a higher prevalence in HIV-positive patients. The specific role that these viruses play in the pathogenesis of apical periodontitis remains to be described.

  2. Plant Origin of Green Propolis: Bee Behavior, Plant Anatomy and Chemistry

    PubMed Central

    2005-01-01

    Propolis, a honeybee product, has gained popularity as a food and alternative medicine. Its constituents have been shown to exert pharmacological effects, such as anti-microbial, anti-inflammatory and anticancer. Shoot apices of Baccharis dracunculifolia (alecrim plant, Asteraceae) have been pointed out as sources of resin for green propolis. The present work aimed (i) to observe the collecting behavior of bees, (ii) to test the efficacy of histological analysis in studies of propolis botanical origin and (iii) to compare the chemistries of alecrim apices, resin masses and green propolis. Bee behavior was observed, and resin and propolis were microscopically analyzed by inclusion in methacrylate. Ethanol extracts of shoot apices, resin and propolis were analyzed by gas chromatography/mass spectroscopy. Bees cut small fragments from alecrim apices, manipulate and place the resulting mass in the corbiculae. Fragments were detected in propolis and identified as alecrim vestiges by detection of alecrim structures. Prenylated and non-prenylated phenylpropanoids, terpenoids and compounds from other classes were identified. Compounds so far unreported for propolis were identified, including anthracene derivatives. Some compounds were found in propolis and resin mass, but not in shoot apices. Differences were detected between male and female apices and, among apices, resin and propolis. Alecrim apices are resin sources for green propolis. Chemical composition of alecrim apices seems to vary independently of season and phenology. Probably, green propolis composition is more complex and unpredictable than previously assumed. PMID:15841282

  3. Ecology of the microbiome of the infected root canal system: a comparison between apical and coronal root segments

    PubMed Central

    Özok, A.R.; Persoon, I.F.; Huse, S.M.; Keijser, B.J.F.; Wesselink, P.R.; Crielaard, W.; Zaura, E.

    2016-01-01

    Aim To evaluate the microbial ecology of the coronal and apical segments of infected root canal systems using a complete sampling technique and next-generation sequencing. Methodology The roots of 23 extracted teeth with apical periodontitis were sectioned in half, horizontally, and cryo-pulverized. Bacterial communities were profiled using tagged 454 pyrosequencing of the 16S rDNA hypervariable V5–V6 region. Results The sequences were classified into 606 taxa (species or higher taxon), representing 24 bacterial phyla or candidate divisions and one archaeal phylum. Proteobacteria were more abundant in the apical samples (p<0.05), while Actinobacteria were in significantly higher proportions in the coronal samples. The apical samples harbored statistically significantly more taxa than the coronal samples (p=0.01), and showed a higher microbial diversity. Several taxa belonging to fastidious obligate anaerobes were significantly more abundant in the apical segments of the roots compared to their coronal counterparts. Conclusions Endodontic infections are more complex than reported previously. The apical part of the root canal system drives the selection of a more diverse and more anaerobe community than the coronal part. The presence of a distinct ecological niche in the apical region explains the difficulty of eradication of the infection, and emphasizes the need that new treatment approaches should be developed. PMID:22251411

  4. Does apical negative pressure prevent the apical extrusion of debris and irrigant compared with conventional irrigation? A systematic review and meta-analysis.

    PubMed

    Romualdo, Priscilla Coutinho; de Oliveira, Katharina Morant Holanda; Nemezio, Mariana Alencar; Küchler, Erika Calvano; Silva, Raquel Assed Bezerra; Nelson-Filho, Paulo; Silva, Lea Assed Bezerra

    2017-12-01

    The aim of this study was to evaluate if apical negative pressure (ANP) irrigation prevents the apical extrusion of debris and irrigant compared with conventional needle irrigation through a systematic review and meta-analysis. A computer search of dental literature was performed using four different databases. A combination of the terms 'apical negative pressure', 'endovac', 'apical extrusion', 'extrusion' and 'endodontics' was used. Studies that used extracted human teeth with a mature apex and that evaluated the apical extrusion of debris and/or irrigating solution were included. After an evaluation of the full studies according to the eligibility criteria, eight studies were critically analysed and subjected to quality assessment and risk of bias. Only four studies that evaluated extrusion of irrigant were considered as having high methodological quality and were subjected to a meta-analysis. Studies evaluating extrusion of debris did not have sufficient methodological quality to be subjected to the meta-analysis. The forest plot indicated that ANP irrigation prevents the risk of irrigant extrusion compared with conventional irrigation (OR 0.07 [95%CI 0.02-0.20]; P < 0.00001). This systematic review and meta-analysis showed that ANP prevents the apical extrusion of irrigant. There is no evidence if this type of irrigation prevents the extrusion of debris. © 2017 Australian Society of Endodontology Inc.

  5. Apical extrusion of debris and irrigants using two hand and three engine-driven instrumentation techniques.

    PubMed

    Ferraz, C C; Gomes, N V; Gomes, B P; Zaia, A A; Teixeira, F B; Souza-Filho, F J

    2001-07-01

    To evaluate the weight of debris and irrigant volume extruded apically from extracted teeth in vitro after endodontic instrumentation using the balanced force technique, a hybrid hand instrumentation technique, and three engine-driven techniques utilizing nickel-titanium instruments (ProFile .04, Quantec 2000 and Pow-R). Five groups of 20 extracted human teeth with single canals were instrumented using one or other of five techniques: balanced force, hybrid, Quantec 2000, ProFile .04, or Pow-R. Debris extruded from the apical foramen during instrumentation were collected into preweighed 1.5 mL tubes. Following instrumentation, the volume of extruded irrigant fluid was determined by visual comparison to control centrifuge tubes filled with 0.25 mL increments of distilled water. The weight of dry extruded dentine debris was also established. Overall, the engine-driven techniques extruded less debris than the manual ones. However, there was no statistical difference between the balanced force technique and the engine-driven methods. The volume of irrigant extruded through the apex was directly associated with the weight of extruded debris, except within the ProFile group. The hybrid technique was associated with the greatest extrusion of both debris and irrigant. Overall, the engine-driven nickel-titanium systems were associated with less apical extrusion.

  6. An ex vivo quantification of the apically extruded bacteria following use of nickel-titanium rotary instruments.

    PubMed

    Mohammadi, Zahed

    2007-01-01

    The aim of this study was to evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using the five different engine-driven techniques utilizing nickel-titanium instruments. One-hundred and ten extracted maxillary central incisor teeth were used. Access cavities were prepared, and root canals were then contaminated with a suspension of Enterococcus faecalis and dried. The contaminated roots were divided into five experimental groups of 20 teeth each and one control group of 10 teeth: Group 1, HERO 642; Group 2, Flex Master; Group 3, Profile GT; Group 4, RoCe; Group 5, K3; and Group 6, control group. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 hours. Colonies of bacteria were counted, and the results were given as the number of colony-forming units. The data obtained were analyzed using one-way ANOVA, with p = 0.05 as the level for statistical significance. There was no significant difference as to the number of extruded bacteria between five engine-driven systems (p > 0.05). All engine-driven nickel-titanium systems extruded bacteria through the apical foramen.

  7. Ex vivo study on root canal instrumentation of two rotary nickel-titanium systems in comparison to stainless steel hand instruments.

    PubMed

    Vaudt, J; Bitter, K; Neumann, K; Kielbassa, A M

    2009-01-01

    To investigate instrumentation time, working safety and the shaping ability of two rotary nickel-titanium (NiTi) systems (Alpha System and ProTaper Universal) in comparison to stainless steel hand instruments. A total of 45 mesial root canals of extracted human mandibular molars were selected. On the basis of the degree of curvature the matched teeth were allocated randomly into three groups of 15 teeth each. In group 1 root canals were prepared to size 30 using a standardized manual preparation technique; in group 2 and 3 rotary NiTi instruments were used following the manufacturers' instructions. Instrumentation time and procedural errors were recorded. With the aid of pre- and postoperative radiographs, apical straightening of the canal curvature was determined. Photographs of the coronal, middle and apical cross-sections of the pre- and postoperative canals were taken, and superimposed using a standard software. Based on these composite images the portion of uninstrumented canal walls was evaluated. Active instrumentation time of the Alpha System was significantly reduced compared with ProTaper Universal and hand instrumentation (P < 0.05; anova). No instrument fractures occurred in any of the groups. The Alpha System revealed significantly less apical straightening compared with the other instruments (P < 0.05; Mann-Whitney U test). In the apical cross-sections Alpha System resulted in significantly less uninstrumented canal walls compared with stainless steel files (P < 0.05; chi-squared test). Despite the demonstrated differences between the systems, an apical straightening effect could not be prevented; areas of uninstrumented root canal wall were left in all regions using the various systems.

  8. Effect of Reciprocating Systems and Working Lengths on Apical Microcrack Development: a micro-CT Study.

    PubMed

    Oliveira, Bruna Paloma de; Câmara, Andréa Cruz; Duarte, Daniel Amancio; Heck, Richard John; Antonino, Antonio Celso Dantas; Aguiar, Carlos Menezes

    2017-01-01

    The objective of this study was to evaluate the effect of root canal preparation with single-file reciprocating systems at different working lengths on the development of apical microcracks using micro-computed tomographic (micro-CT) imaging. Forty extracted human mandibular incisors were randomly assigned to 4 groups (n=10) according to the systems and working length used to prepare the root canals: Group A - WaveOne Gold at apical foramen (AF), Group B - WaveOne Gold 1 mm short of the AF (AF-1 mm), Group C - Unicone (AF) and Group D - Unicone (AF-1 mm). Micro-CT scanning was performed before and after root canal preparation at an isotropic resolution of 14 µm. Then, three examiners assessed the cross-sectional images generated to detect microcracks in the apical portion of the roots. Apical microcracks were visualized in 3, 1, 1, and 3 specimens in groups A, B, C, and D, respectively. All these microcracks observed after root canal preparation already existed prior to instrumentation, and no new apical microcrack was detected. For all groups, the number of slices presenting microcracks after root canal preparation was the same as before canal preparation. Root canal preparation with WaveOne Gold and Unicone, regardless of the working length, was not associated with apical microcrack formation.

  9. Pak1 Kinase Maintains Apical Membrane Identity in Epithelia.

    PubMed

    Aguilar-Aragon, Mario; Elbediwy, Ahmed; Foglizzo, Valentina; Fletcher, Georgina C; Li, Vivian S W; Thompson, Barry J

    2018-02-13

    Epithelial cells are polarized along their apical-basal axis by the action of the small GTPase Cdc42, which is known to activate the aPKC kinase at the apical domain. However, loss of aPKC kinase activity was reported to have only mild effects on epithelial cell polarity. Here, we show that Cdc42 also activates a second kinase, Pak1, to specify apical domain identity in Drosophila and mammalian epithelia. aPKC and Pak1 phosphorylate an overlapping set of polarity substrates in kinase assays. Inactivating both aPKC kinase activity and the Pak1 kinase leads to a complete loss of epithelial polarity and morphology, with cells losing markers of apical polarization such as Crumbs, Par3/Bazooka, or ZO-1. This function of Pak1 downstream of Cdc42 is distinct from its role in regulating integrins or E-cadherin. Our results define a conserved dual-kinase mechanism for the control of apical membrane identity in epithelia. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

  10. Electrogenic active proton pump in Rana esculenta skin and its role in sodium ion transport.

    PubMed

    Ehrenfeld, J; Garcia-Romeu, F; Harvey, B J

    1985-02-01

    Kinetic and electrophysiological studies were carried out in the in vitro Rana esculenta skin, bathed in dilute sodium solution, to characterize the proton pump and coupling between sodium absorption (JNa+n) and proton excretion (JH+n). JNa+n and JH+n were both dependent on transepithelial potential (psi ms); hyperpolarizing the skin decreased JNa+n and increased JH+n; depolarization produced the opposite effects. Amiloride (5 X 10(-5) M) at a clamped psi ms of +50 mV inhibited JNa+n without affecting JH+n. Variations of psi ms or pH had identical effects on JH+n. Ethoxzolamide inhibited JH+n and simultaneously increased psi ms by 15-30 mV. These changes were accompanied by depolarization of the apical membrane potential psi mc from -47 to -25 mV and an increase in apical membrane resistance of 30%; no significant effects on basolateral membrane potential (psi cs) and resistance (Rb) nor on shunt resistance (Rj) were observed. The proton pump appears to be localized at the apical membrane. The proton pump was also inhibited by deoxygenation, oligomycin, dicyclohexylcarbodiimide and vanadate (100, 78, 83 and 100% inhibition respectively). The variations of JH+n and of the measured electrical currents were significantly correlated. These findings are supportive evidence of a primary active proton pump, electrogenic and strictly linked to aerobic metabolism. The current-voltage (I-V) relation of the proton pump was obtained as the difference in the I-V curves of the apical membrane extracted before and after proton-pump inhibition by ethoxzolamide during amiloride block of sodium transport. The proton-pump current (IP) was best described by a saturable exponential function of psi mc. Maximal pump current (ImaxP) was calculated to be 200 nequiv h-1 cm-2 at a psi mc of +50 mV and the pump reversal potential ERP was -130 mV. The effect of ethoxzolamide to depolarize psi mc was dependent on the relation between psi mc and ERP. Maximal induced depolarization occurred at a psi mc of +50 mV whereas ethoxzolamide exerted minimal effect on psi mc when the ERP was approached either by voltage clamping the apical membrane or by the addition of amiloride. We show that electroneutral sodium-proton countertransport is not the mechanism of active proton excretion in frog skin but that it is the proton excretion which provides a favourable electrical driving force for passive apical sodium entry.(ABSTRACT TRUNCATED AT 400 WORDS)

  11. Electrogenic active proton pump in Rana esculenta skin and its role in sodium ion transport.

    PubMed Central

    Ehrenfeld, J; Garcia-Romeu, F; Harvey, B J

    1985-01-01

    Kinetic and electrophysiological studies were carried out in the in vitro Rana esculenta skin, bathed in dilute sodium solution, to characterize the proton pump and coupling between sodium absorption (JNa+n) and proton excretion (JH+n). JNa+n and JH+n were both dependent on transepithelial potential (psi ms); hyperpolarizing the skin decreased JNa+n and increased JH+n; depolarization produced the opposite effects. Amiloride (5 X 10(-5) M) at a clamped psi ms of +50 mV inhibited JNa+n without affecting JH+n. Variations of psi ms or pH had identical effects on JH+n. Ethoxzolamide inhibited JH+n and simultaneously increased psi ms by 15-30 mV. These changes were accompanied by depolarization of the apical membrane potential psi mc from -47 to -25 mV and an increase in apical membrane resistance of 30%; no significant effects on basolateral membrane potential (psi cs) and resistance (Rb) nor on shunt resistance (Rj) were observed. The proton pump appears to be localized at the apical membrane. The proton pump was also inhibited by deoxygenation, oligomycin, dicyclohexylcarbodiimide and vanadate (100, 78, 83 and 100% inhibition respectively). The variations of JH+n and of the measured electrical currents were significantly correlated. These findings are supportive evidence of a primary active proton pump, electrogenic and strictly linked to aerobic metabolism. The current-voltage (I-V) relation of the proton pump was obtained as the difference in the I-V curves of the apical membrane extracted before and after proton-pump inhibition by ethoxzolamide during amiloride block of sodium transport. The proton-pump current (IP) was best described by a saturable exponential function of psi mc. Maximal pump current (ImaxP) was calculated to be 200 nequiv h-1 cm-2 at a psi mc of +50 mV and the pump reversal potential ERP was -130 mV. The effect of ethoxzolamide to depolarize psi mc was dependent on the relation between psi mc and ERP. Maximal induced depolarization occurred at a psi mc of +50 mV whereas ethoxzolamide exerted minimal effect on psi mc when the ERP was approached either by voltage clamping the apical membrane or by the addition of amiloride. We show that electroneutral sodium-proton countertransport is not the mechanism of active proton excretion in frog skin but that it is the proton excretion which provides a favourable electrical driving force for passive apical sodium entry.(ABSTRACT TRUNCATED AT 400 WORDS) Images Fig. 6 Fig. 7 PMID:2582114

  12. Soybean extracts increase cell surface ZIP4 abundance and cellular zinc levels: a potential novel strategy to enhance zinc absorption by ZIP4 targeting.

    PubMed

    Hashimoto, Ayako; Ohkura, Katsuma; Takahashi, Masakazu; Kizu, Kumiko; Narita, Hiroshi; Enomoto, Shuichi; Miyamae, Yusaku; Masuda, Seiji; Nagao, Masaya; Irie, Kazuhiro; Ohigashi, Hajime; Andrews, Glen K; Kambe, Taiho

    2015-12-01

    Dietary zinc deficiency puts human health at risk, so we explored strategies for enhancing zinc absorption. In the small intestine, the zinc transporter ZIP4 functions as an essential component of zinc absorption. Overexpression of ZIP4 protein increases zinc uptake and thereby cellular zinc levels, suggesting that food components with the ability to increase ZIP4 could potentially enhance zinc absorption via the intestine. In the present study, we used mouse Hepa cells, which regulate mouse Zip4 (mZip4) in a manner indistinguishable from that in intestinal enterocytes, to screen for suitable food components that can increase the abundance of ZIP4. Using this ZIP4-targeting strategy, two such soybean extracts were identified that were specifically able to decrease mZip4 endocytosis in response to zinc. These soybean extracts also effectively increased the abundance of apically localized mZip4 in transfected polarized Caco2 and Madin-Darby canine kidney cells and, moreover, two apically localized mZip4 acrodermatitis enteropathica mutants. Soybean components were purified from one extract and soyasaponin Bb was identified as an active component that increased both mZip4 protein abundance and zinc levels in Hepa cells. Finally, we confirmed that soyasaponin Bb is capable of enhancing cell surface endogenous human ZIP4 in human cells. Our results suggest that ZIP4 targeting may represent a new strategy to improve zinc absorption in humans. © 2015 Authors; published by Portland Press Limited.

  13. [Developmental radicular groove as a cause of endodontic failure].

    PubMed

    Fabra Campos, H; Millet Part, J

    1989-01-01

    A clinical case of apical injury on an upper lateral incisor with endodontical and surgical failures in its treatment is presented. Extraction of the incisor and its study at the stereoscopic microscope showed the existence of a developmental groove running from the cingulum to the end of the root, establishing a communication between the crevice and the apical part of the tooth. Bacterial infection through the groove could provide an explanation for treatment failure.

  14. Effect of glide path and apical preparation size on the incidence of apical crack during the canal preparation using Reciproc, WaveOne, and ProTaper Next systems in curved root canals: A stereomicroscope study.

    PubMed

    Topçuoğlu, Hüseyin Sinan; Düzgün, Salih; Akpek, Firdevs; Topçuoğlu, Gamze

    2016-11-01

    This study evaluated the effect of creating a glide path and apical preparation size on the incidence of apical cracks during canal preparation in mandibular molar teeth with curved canals. One hundred and forty extracted teeth were used. The teeth were randomly assigned to one control group or six experimental groups (n = 20 per group) for canal preparation. No preparation was performed on teeth in the control group. In three of the six experimental groups, a glide path was not created; a glide path was created on the curved mesial canals of all teeth in the remaining three experimental groups. All teeth in experimental groups were then instrumented with the following systems: Reciproc, WaveOne (WO), and ProTaper Next (PTN). Digital images of the apical root surfaces of these teeth were recorded before preparation, after instrumentation with size 25 files, and after instrumentation with size 40 files. The images were then inspected for the presence of any new apical cracks and propagation. There was no significant difference between the experimental groups during canal preparation using size 25 files (p > 0.05). Reciproc and WO caused more new apical cracks than did PTN during canal preparation using size 40 files (p < 0.05). However, canal preparation using size 40 files did not cause propagation of existing cracks (p > 0.05). Performing a glide path prior to canal preparation did not change the incidence of apical crack during preparation. Additionally, increasing apical preparation size may increase the incidence of apical crack during canal preparation. SCANNING 38:585-590, 2016. © 2016 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

  15. The effect of cervical preflaring using different rotary nickel-titanium systems on the accuracy of apical file size determination.

    PubMed

    Tennert, Christian; Herbert, Josef; Altenburger, Markus Jörg; Wrbas, Karl-Thomas

    2010-10-01

    An exact determination of the apical root canal diameter is crucial for correct cleaning and shaping of a root canal. The aim of this study was to investigate the discrepancies of the initial apical root canal diameter and the diameter that is measured by the initial apical file (IAF) after cervical flaring using current rotary nickel-titanium systems. Mesiobuccal canals of 40 extracted mandibular molars were randomly assigned to four groups. In the first group, root canals were not flared. Root canals of the other groups were preflared using FlexMaster (VDW, Munich, Germany), ProTaper (Dentsply, Konstanz, Germany), or RaCe (FKG Dentaire, Genf, Switzerland) instruments. The tooth length was determined by inserting an ISO 06 K-file to the apical foramen. The working length (WL) was set 1 mm short of the apical foramen. File sizes were increased after binding sensation was felt at the WL. Transversal sections of the WL regions were examined under stereomicroscope, and the diameter of the root canal and the IAF at WL were assessed. Canals preflared with RaCe instruments had the lowest discrepancy between the apical root canal diameter and the IAF diameter (15.7 ± 9.7 μm) followed by ProTaper (22.2 ± 11.0 μm) and FlexMaster (35.0 ± 17.2 μm). Preflaring of root canals prevents underestimation of the actual apical root canal diameter. The type of instruments used for preflaring show differences on the accuracy of IAF determination. Preflaring with larger tapered instruments leads to a more accurate apical sizing, and this information is crucial concerning the appropriate final diameter for complete apical shaping. Copyright © 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  16. Association analysis of clinical aspects and vitamin D receptor gene polymorphism with external apical root resorption in orthodontic patients.

    PubMed

    Fontana, Maria Luiza S Simas Netta; de Souza, Cleber Machado; Bernardino, José Fabio; Hoette, Felix; Hoette, Maura Levi; Thum, Lotario; Ozawa, Terumi O; Capelozza Filho, Leopoldino; Olandoski, Marcia; Trevilatto, Paula Cristina

    2012-09-01

    Vitamin D is responsible for the regulation of certain genes at the transcription level, via interaction with the vitamin D receptor, and influences host immune responses and aspects of bone development, growth, and homeostasis. Our aim was to investigate the association of TaqI vitamin D receptor gene polymorphism with external apical root resorption during orthodontic treatment. Our subjects were 377 patients with Class II Division 1 malocclusion, divided into 3 groups: (1) 160 with external apical root resorption ≤1.43 mm, (2) 179 with external apical root resorption >1.43 mm), and (3) 38 untreated subjects. External apical root resorption of the maxillary incisors was evaluated on periapical radiographs taken before and after 6 months of treatment. After DNA collection and purification, vitamin D receptor TaqI polymorphism analysis was performed by polymerase chain reaction-restriction fragment length polymorphism. Univariate and multivariate analyses were performed to verify the association of clinical and genetic variables with external apical root resorption (P <0.05). There was a higher proportion of external apical root resorption in orthodontically treated patients compared with the untreated subjects. In patients orthodontically treated, age higher than 14 years old, initial size of the maxillary incisor root superior to 30 mm, and premolar extraction were associated with increased external apical root resorption. Genotypes containing the C allele were weakly associated with protection against external apical root resorption (CC + CT × TT [odds ratio, 0.29; 95% confidence interval, 0.07-1.23; P = 0.091]) when treated orthodontic patients were compared to untreated individuals. Clinical factors and vitamin D receptor TaqI polymorphism were associated with external apical root resorption in orthodontic patients. Copyright © 2012 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.

  17. Quantitative assessment of apical debris extrusion and intracanal debris in the apical third, using hand instrumentation and three rotary instrumentation systems.

    PubMed

    H K, Sowmya; T S, Subhash; Goel, Beena Rani; T N, Nandini; Bhandi, Shilpa H

    2014-02-01

    Decreased apical extrusion of debris and apical one third debris have strong implications for decreased incidence of postoperative inflammation and pain. Thus, the aim of this study was to assess quantitatively the apical extrusion of debris and intracanal debris in the apical third during root canal instrumentation using hand and three different types of rotary instruments. Sixty freshly extracted single rooted human teeth were randomly divided into four groups. Canal preparation was done using step-back with hand instrumentation, crown-down technique with respect to ProTaper and K3, and hybrid technique with LightSpeed LSX. Irrigation was done with NaOCl, EDTA, and normal saline and for final irrigation, EndoVac system was used. The apically extruded debris was collected on the pre-weighed Millipore plastic filter disk and weighed using microbalance. The teeth were submitted to the histological processing. Sections from the apical third were analyzed by a trinocular research microscope that was coupled to a computer where the images were captured and analyzed using image proplus V4.1.0.0 software. The mean weight of extruded debris for each group and intracanal debris in the root canal was statistically analyzed by a Kruskal-Wallis one-way analysis of variance and Mann-Whitney U test. The result showed that, hand instrumentation using K files showed the highest amount of debris extrusion apically when compared to ProTaper, K3 and LightSpeed LSX. The result also showed that there was no statistically significant difference between the groups in relation to presence of intracanal debris in the apical one third. Based on the results, all instrumentation techniques produced debris extrusion. The engine driven Ni-Ti systems extruded significantly less apical debris than hand instrumentation. There was no statistically significant difference between the groups in relation to presence of intracanal debris in the apical one third.

  18. Quantitative Assessment of Apical Debris Extrusion and Intracanal Debris in the Apical Third, Using Hand Instrumentation and Three Rotary Instrumentation Systems

    PubMed Central

    H.K., Sowmya; T.S., Subhash; Goel, Beena Rani; T.N., Nandini; Bhandi, Shilpa H.

    2014-01-01

    Introduction: Decreased apical extrusion of debris and apical one third debris have strong implications for decreased incidence of postoperative inflammation and pain. Thus, the aim of this study was to assess quantitatively the apical extrusion of debris and intracanal debris in the apical third during root canal instrumentation using hand and three different types of rotary instruments. Methodology: Sixty freshly extracted single rooted human teeth were randomly divided into four groups. Canal preparation was done using step-back with hand instrumentation, crown-down technique with respect to ProTaper and K3, and hybrid technique with LightSpeed LSX. Irrigation was done with NaOCl, EDTA, and normal saline and for final irrigation, EndoVac system was used. The apically extruded debris was collected on the pre-weighed Millipore plastic filter disk and weighed using microbalance. The teeth were submitted to the histological processing. Sections from the apical third were analyzed by a trinocular research microscope that was coupled to a computer where the images were captured and analyzed using image proplus V4.1.0.0 software. The mean weight of extruded debris for each group and intracanal debris in the root canal was statistically analyzed by a Kruskal-Wallis one-way analysis of variance and Mann-Whitney U test. Results: The result showed that, hand instrumentation using K files showed the highest amount of debris extrusion apically when compared to ProTaper, K3 and LightSpeed LSX. The result also showed that there was no statistically significant difference between the groups in relation to presence of intracanal debris in the apical one third. Conclusion: Based on the results, all instrumentation techniques produced debris extrusion. The engine driven Ni-Ti systems extruded significantly less apical debris than hand instrumentation. There was no statistically significant difference between the groups in relation to presence of intracanal debris in the apical one third. PMID:24701536

  19. Increased Na+/H+ exchanger activity on the apical surface of a cilium-deficient cortical collecting duct principal cell model of polycystic kidney disease

    PubMed Central

    Olteanu, Dragos; Liu, Xiaofen; Liu, Wen; Roper, Venus C.; Sharma, Neeraj; Yoder, Bradley K.; Satlin, Lisa M.; Schwiebert, Erik M.

    2012-01-01

    Pathophysiological anomalies in autosomal dominant and recessive forms of polycystic kidney disease (PKD) may derive from impaired function/formation of the apical central monocilium of ductal epithelia such as that seen in the Oak Ridge polycystic kidney or orpk (Ift88Tg737Rpw) mouse and its immortalized cell models for the renal collecting duct. According to a previous study, Na/H exchanger (NHE) activity may contribute to hyperabsorptive Na+ movement in cilium-deficient (“mutant”) cortical collecting duct principal cell monolayers derived from the orpk mice compared with cilium-competent (“rescued”) monolayers. To examine NHE activity, we measured intracellular pH (pHi) by fluorescence imaging with the pH-sensitive dye BCECF, and used a custom-designed perfusion chamber to control the apical and basolateral solutions independently. Both mutant and rescued monolayers exhibited basolateral Na+-dependent acid-base transporter activity in the nominal absence of CO2/HCO3−. However, only the mutant cells displayed appreciable apical Na+-induced pHi recoveries from NH4+ prepulse-induced acid loads. Similar results were obtained with isolated, perfused collecting ducts from orpk vs. wild-type mice. The pHi dependence of basolateral cariporide/HOE-694-sensitive NHE activity under our experimental conditions was similar in both mutant and rescued cells, and 3.5- to 4.5-fold greater than apical HOE-sensitive NHE activity in the mutant cells (pHi 6.23–6.68). Increased apical NHE activity correlated with increased apical NHE1 expression in the mutant cells, and increased apical localization in collecting ducts of kidney sections from orpk vs. control mice. A kidney-specific conditional cilium-knockout mouse produced a more acidic urine compared with wild-type littermates and became alkalotic by 28 days of age. This study provides the first description of altered NHE activity, and an associated acid-base anomaly in any form of PKD. PMID:22301060

  20. Analysis of correlation between initial alveolar bone density and apical root resorption after 12 months of orthodontic treatment without extraction

    PubMed Central

    Scheibel, Paula Cabrini; Ramos, Adilson Luiz; Iwaki, Lilian Cristina Vessoni; Micheletti, Kelly Regina

    2014-01-01

    OBJECTIVE: The aim of the present study was to investigate the correlation between initial alveolar bone density of upper central incisors (ABD-UI) and external apical root resorption (EARR) after 12 months of orthodontic movement in cases without extraction. METHODS: A total of 47 orthodontic patients 11 years old or older were submitted to periapical radiography of upper incisors prior to treatment (T1) and after 12 months of treatment (T2). ABD-UI and EARR were measured by means of densitometry. RESULTS: No statistically significant correlation was found between initial ABD-UI and EARR at T2 (r = 0.149; p = 0.157). CONCLUSION: Based on the present findings, alveolar density assessed through periapical radiography is not predictive of root resorption after 12 months of orthodontic treatment in cases without extraction. PMID:25715722

  1. Prevalence and activity of Epstein-Barr virus and human cytomegalovirus in symptomatic and asymptomatic apical periodontitis lesions.

    PubMed

    Hernádi, Katinka; Szalmás, Anita; Mogyorósi, Richárd; Czompa, Levente; Veress, György; Csoma, Eszter; Márton, Ildikó; Kónya, József

    2010-09-01

    Apical periodontitis is a polymicrobial inflammation with a dominant flora of opportunistic Gram-negative bacteria; however, a pathogenic role of human herpesviruses such as Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) has been implicated recently. The aims of this study were to determine the prevalence, activity, and disease association of EBV and HCMV in apical periodontitis in an Eastern Hungarian population. Forty samples with apical periodontitis (17 symptomatic and 23 asymptomatic) and 40 healthy pulp controls were collected. EBV and HCMV prevalences were measured by polymerase chain reaction (PCR) detection of the viral DNA and viral activity was tested by reverse-transcription PCR amplification of viral messenger RNA. EBV DNA and EBNA-2 messenger RNA were found in apical periodontitis lesions at significantly (p < 0.0001) higher frequencies (72.5% and 50%, respectively) than in controls (both 2.5%). The occurrence of HCMV infection was rare in both apical lesions (10%) and controls (0%). The presence of EBV DNA in apical lesions was associated significantly with large (> or = 5 mm) lesion size (p = 0.02) but not with symptoms (p = 0.30). Symptomatic manifestation was significantly associated with the co-occurrence (odds ratio [OR], 8.80; 95% confidence interval [CI], 1.69-45.76) but not the sole occurrences of EBNA-2 messenger RNA (OR, 2.29; 95% CI, 0.48-11.06) and large lesion size (OR, 4.02; 95% CI, 0.81-19.89). EBV infection is a frequent event in apical periodontitis, whereas the involvement of HCMV still remains to be elucidated. This study showed that symptomatic manifestation was likely to occur if a large-sized apical periodontitis lesion is aggravated with active EBV infection. Copyright 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  2. K(Ca)3.1 channels facilitate K+ secretion or Na+ absorption depending on apical or basolateral P2Y receptor stimulation.

    PubMed

    Palmer, Melissa L; Peitzman, Elizabeth R; Maniak, Peter J; Sieck, Gary C; Prakash, Y S; O'Grady, Scott M

    2011-07-15

    Human mammary epithelial (HME) cells express several P2Y receptor subtypes located in both apical and basolateral membranes. Apical UTP or ATP-γ-S stimulation of monolayers mounted in Ussing chambers evoked a rapid, but transient decrease in short circuit current (I(sc)), consistent with activation of an apical K+ conductance. In contrast, basolateral P2Y receptor stimulation activated basolateral K+ channels and increased transepithelial Na+ absorption. Chelating intracellular Ca2+ using the membrane-permeable compound BAPTA-AM, abolished the effects of purinoceptor activation on I(sc). Apical pretreatment with charybdotoxin also blocked the I(sc) decrease by >90% and similar magnitudes of inhibition were observed with clotrimazole and TRAM-34. In contrast, iberiotoxin and apamin did not block the effects of apical P2Y receptor stimulation. Silencing the expression of K(Ca)3.1 produced ∼70% inhibition of mRNA expression and a similar reduction in the effects of apical purinoceptor agonists on I(sc). In addition, silencing P2Y2 receptors reduced the level of P2Y2 mRNA by 75% and blocked the effects of ATP-γ-S by 65%. These results suggest that P2Y2 receptors mediate the effects of purinoceptor agonists on K+ secretion by regulating the activity of K(Ca)3.1 channels expressed in the apical membrane of HME cells. The results also indicate that release of ATP or UTP across the apical or basolateral membrane elicits qualitatively different effects on ion transport that may ultimately determine the [Na+]/[K+] composition of fluid within the mammary ductal network.

  3. A comparison of residual smear layer and erosion following different endodontic irrigation protocols tested under clinical and laboratory conditions.

    PubMed

    Cehreli, Zafer C; Uyanik, M Ozgur; Nagas, Emre; Tuncel, Behram; Er, Nuray; Comert, Fugen Dagli

    2013-09-01

    To compare the smear layer removal efficacy and erosive effects of different irrigation protocols under clinical and laboratory conditions. Mandibular third molars (n = 32) of 30-45 year-old patients were instrumented with rotary files and were randomly assigned to one of the following groups for final irrigation: (1) 5.25% NaOCl; (2) 17% EDTA; and (3) BioPure MTAD. Thereafter, the teeth were immediately extracted and processed for micromorphological investigation. In vitro specimen pairs were prepared by repeating the clinical experiments on freshly-extracted mandibular third molars. To compare open and closed systems, laboratory experiments were repeated on 32 additional teeth with enlarged apical foramen. The cleanliness of the root canals and the extent of erosion were assessed by environmental scanning electron microscopy. Specimens prepared under clinical and laboratory conditions had similar cleanliness and erosion scores (p > 0.05). Under both conditions, the tested solutions were more effective in removing the smear layer in the coronal and middle regions than in the apical one. Comparison of closed and open systems showed similar levels of cleanliness and erosion in all regions (p > 0.05), with the exception of 17% EDTA showing significantly higher levels of cleanliness and erosion in the apical third of open-end specimens. Based on clinical correlates of in vitro root canal cleanliness and erosion, laboratory testing of root canal irrigants on extracted teeth with closed apices can serve as a reliable method to simulate the clinical condition. EDTA was the most effective final irrigation solution in removing the smear layer at the expense of yielding the greatest erosive effect.

  4. Histology of periapical lesions obtained during apical surgery.

    PubMed

    Schulz, Malte; von Arx, Thomas; Altermatt, Hans Jörg; Bosshardt, Dieter

    2009-05-01

    The aim of this was to evaluate the histology of periapical lesions in teeth treated with periapical surgery. After root-end resection, the root tip was removed together with the periapical pathological tissue. Histologic sectioning was performed on calcified specimens embedded in methylmethacrylate (MMA) and on demineralized specimens embedded in LR White (Fluka, Buchs, Switzerland). The samples were evaluated with light and transmission electron microscopy (TEM). The histologic findings were classified into periapical abscesses, granulomas, or cystic lesions (true or pocket cysts). The final material comprised 70% granulomas, 23% cysts and 5% abscesses, 1% scar tissues, and 1% keratocysts. Six of 125 samples could not be used. The cystic lesions could not be subdivided into pocket or true cysts. All cysts had an epithelium-lined cavity, two of them with cilia-lined epithelium. These results show the high incidence of periapical granulomas among periapical lesions obtained during apical surgery. Periapical abscesses were a rare occasion. The histologic findings from samples obtained during apical surgery may differ from findings obtained by teeth extractions. A determination between pocket and true apical cysts is hardly possible when collecting samples by apical surgery.

  5. Hypercementosis and odontogenic epithelial hyperplasia associated with a tooth root remnant mimicking a neoplasm. A case report.

    PubMed

    Zustin, J; Friedrich, R E

    2010-01-01

    Hypercementosis presents as painless, single or multiple non-neoplastic cementum formation beyond the physiological limits of the tooth. It often occurs in the apical area of the involved tooth following infection, chemical or mechanical trauma. We report on radiographic and histopathological findings in a single case of late intraosseous hypercementosis and odontogenic epithelial hyperplasia associated with a minute apical tooth root remnant years after its extraction, mimicking a tumour.

  6. Apical extrusion of intracanal bacteria following use of two engine-driven instrumentation techniques.

    PubMed

    Er, K; Sümer, Z; Akpinar, K E

    2005-12-01

    To evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using the two engine-driven techniques utilizing nickel-titanium instruments (ProTaper and System GT). Forty extracted single-rooted human mandibular premolar teeth were used. Access cavities were prepared and root canals were then contaminated with a suspension of Enterococcus faecalis and dried. The contaminated roots were divided into two experimental groups of 15 teeth each and one control group of 10 teeth. Group 1, ProTaper group: the root canals were instrumented using ProTaper instruments. Group 2, System GT group: the root canals were instrumented using System GT instruments. Group 3, control group: no instrumentation was attempted. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 h. Colonies of bacteria were counted and the results were given as number of colony-forming units. The data obtained were analysed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests, with alpha = 0.05 as the level for statistical significance. There was no significant difference as to the number of extruded bacteria between the ProTaper and System GT engine-driven systems (P > 0.05). Both engine-driven nickel-titanium systems extruded bacteria through the apical foramen.

  7. Src- and Fyn-dependent apical membrane trafficking events control endothelial lumen formation during vascular tube morphogenesis.

    PubMed

    Kim, Dae Joong; Norden, Pieter R; Salvador, Jocelynda; Barry, David M; Bowers, Stephanie L K; Cleaver, Ondine; Davis, George E

    2017-01-01

    Here we examine the question of how endothelial cells (ECs) develop their apical membrane surface domain during lumen and tube formation. We demonstrate marked apical membrane targeting of activated Src kinases to this apical domain during early and late stages of this process. Immunostaining for phosphotyrosine or phospho-Src reveals apical membrane staining in intracellular vacuoles initially. This is then followed by vacuole to vacuole fusion events to generate an apical luminal membrane, which is similarly decorated with activated phospho-Src kinases. Functional blockade of Src kinases completely blocks EC lumen and tube formation, whether this occurs during vasculogenic tube assembly or angiogenic sprouting events. Multiple Src kinases participate in this apical membrane formation process and siRNA suppression of Src, Fyn and Yes, but not Lyn, blocks EC lumen formation. We also demonstrate strong apical targeting of Src-GFP and Fyn-GFP fusion proteins and increasing their expression enhances lumen formation. Finally, we show that Src- and Fyn-associated vacuoles track and fuse along a subapically polarized microtubule cytoskeleton, which is highly acetylated. These vacuoles generate the apical luminal membrane in a stereotypically polarized, perinuclear position. Overall, our study identifies a critical role for Src kinases in creating and decorating the EC apical membrane surface during early and late stages of lumen and tube formation, a central event in the molecular control of vascular morphogenesis.

  8. A radiographic study of external apical root resorption in patients treated with single-phase fixed orthodontic therapy.

    PubMed

    Agarwal, S S; Chopra, S S; Kumar, Prasanna; Jayan, B; Nehra, K; Sharma, Mohit

    2016-12-01

    External apical root resorption (EARR) is one of the most common iatrogenic consequences of orthodontic tooth movement. Many factors like gender, duration, orthodontic force and duration of orthodontic treatment have been implicated to cause EARR. Pre- and post-treatment OPGs of 60 orthodontic patients (30 males and 30 females) who had undergone treatment with a single phase of fixed orthodontic therapy were randomly selected from institutional archives. The root apices were evaluated for EARR by a single operator on an radiograph viewing box at a standardized source of light using a four-grade ordinal scale. Anterior EARR was measured on the maxillary and mandibular canines. Posterior EARR was measured on premolars, mesiobuccal and distobuccal roots of maxillary first molars and mesial and distal roots of mandibular first molars. The results were compiled and subjected to statistical analysis. The cases in which the patients underwent therapeutic extraction had a relatively higher amount of EARR compared to the cases in which the patients were treated by non-extraction therapy ( P  < 0.001). Odds ratio indicated that extraction cases had two-fold increased risk of EARR than non-extraction cases ( P  < 0.001). No statistically significant difference was observed in the distribution of EARR based on gender or duration of orthodontic treatment ( P  > 0.05). Therapeutic extraction is an important determinant of post-treatment EARR. Gender and duration of orthodontic treatment may not be important variables in the causation of EARR according to the findings of this study. However, longitudinal studies with larger sample size are required to validate the results of this study.

  9. A Comparison of Apical Transportation in Severely Curved Canals Induced by Reciproc and BioRaCe Systems.

    PubMed

    Nabavizadeh, Mohammadreza; Abbaszadegan, Abbas; Khojastepour, Leila; Amirhosseini, Mohsen; Kiani, Ebrahim

    2014-01-01

    Preserving the apical root structure during cleaning and shaping of the canal has always been a challenge in endodontics particularly when the root canals are curved. The purpose of this in vitro study was to compare the apical transportation induced by the Reciproc and BioRaCe rotary systems in preparing the mesiobuccal root canal of the human maxillary molars. The mesiobuccal canals of sixty extracted maxillary molars with curvature angle of 25˚-35˚ were selected and randomly assigned into two groups. Each canal was prepared by either Reciproc or BioRaCe rotary systems. A double-digital radiographic technique and AutoCAD software were used to compare the apical transportation at 0.5, 1, 2, 3, 4 and 5 mm distances from the working length (WL). The distance between the master apical rotary file and the initial K-file in the superimposed radiographs determined the amount of apical transportation. An independent t-test was used to compare the groups. The statistical significant level was set at 0.05. Apical transportation of the Reciproc group was significantly greater than the BioRaCe group in all distances (P<0.001). The maximum apical transportation occurred in the Reciproc group at 0.5 mm from the WL (0.048±0.0028 mm) and the minimum occurred for BioRaCe at 5 mm from the WL (0.010±0.0005 mm). The Reciproc system produced significantly more apical transportation than the BioRaCe, but this fact does not seem to negatively alter the clinical success or quality of root canal treatment.

  10. Apical pressure created during irrigation with the GentleWave™ system compared to conventional syringe irrigation.

    PubMed

    Haapasalo, Markus; Shen, Ya; Wang, Zhejun; Park, Ellen; Curtis, Allison; Patel, Payal; Vandrangi, Prashanthi

    2016-09-01

    The purpose of this study is to compare pressures at the apical foramen created by conventional syringe irrigation and the GentleWave™ System, which releases high-velocity degassed irrigants to the pulp chamber and uses broad-spectrum sound energy for cleaning. The apical pressure generated during irrigation was measured for palatal and distobuccal root canals of four extracted maxillary molars after no instrumentation, minimal instrumentation to a size #15/.04, instrumentation to a size #40/.04 taper, and after perforating the apical foramen to size #40. The root canals opened into an air-tight custom fixture coupled to a piezoresistive pressure transducer. Apical pressures were measured for the GentleWave™ System and syringe-needle irrigation at different irrigant flow rates, with the needle tip at 1 and 3 mm from the apical foramen using 30-gauge (G) open-ended or side-vented safety tip needles. The GentleWave™ System generated negative apical pressures (P < 0.001 compared with syringe irrigation); the mean pressures were between -13.07 and -17.19 mmHg. The 30 G needles could not reach the 1 and 3 mm from the working length in uninstrumented and 1 mm in minimally instrumented canals. The mean positive pressures between 6.46 and 110.34 mmHg were measured with needle irrigation depending on the flow rate, needle insertion depth, and size of the root canal. The GentleWave™ System creates negative pressure at the apical foramen during root canal cleaning irrespective of the size of canal instrumentation. Positive apical pressures were measured for syringe irrigation. Negative pressure during irrigation contributes to improved safety as compared to high-positive pressure.

  11. A Comparison of Apical Transportation in Severely Curved Canals Induced by Reciproc and BioRaCe Systems

    PubMed Central

    Nabavizadeh, Mohammadreza; Abbaszadegan, Abbas; Khojastepour, Leila; Amirhosseini, Mohsen; Kiani, Ebrahim

    2014-01-01

    Introduction: Preserving the apical root structure during cleaning and shaping of the canal has always been a challenge in endodontics particularly when the root canals are curved. The purpose of this in vitro study was to compare the apical transportation induced by the Reciproc and BioRaCe rotary systems in preparing the mesiobuccal root canal of the human maxillary molars. Materials and Methods: The mesiobuccal canals of sixty extracted maxillary molars with curvature angle of 25˚-35˚ were selected and randomly assigned into two groups. Each canal was prepared by either Reciproc or BioRaCe rotary systems. A double-digital radiographic technique and AutoCAD software were used to compare the apical transportation at 0.5, 1, 2, 3, 4 and 5 mm distances from the working length (WL). The distance between the master apical rotary file and the initial K-file in the superimposed radiographs determined the amount of apical transportation. An independent t-test was used to compare the groups. The statistical significant level was set at 0.05. Results: Apical transportation of the Reciproc group was significantly greater than the BioRaCe group in all distances (P<0.001). The maximum apical transportation occurred in the Reciproc group at 0.5 mm from the WL (0.048±0.0028 mm) and the minimum occurred for BioRaCe at 5 mm from the WL (0.010±0.0005 mm). Conclusions: The Reciproc system produced significantly more apical transportation than the BioRaCe, but this fact does not seem to negatively alter the clinical success or quality of root canal treatment. PMID:24688580

  12. AMP-activated protein kinase (AMPK)-dependent and -independent pathways regulate hypoxic inhibition of transepithelial Na+ transport across human airway epithelial cells.

    PubMed

    Tan, C D; Smolenski, R T; Harhun, M I; Patel, H K; Ahmed, S G; Wanisch, K; Yáñez-Muñoz, R J; Baines, D L

    2012-09-01

    Pulmonary transepithelial Na(+) transport is reduced by hypoxia, but in the airway the regulatory mechanisms remain unclear. We investigated the role of AMPK and ROS in the hypoxic regulation of apical amiloride-sensitive Na(+) channels and basolateral Na(+) K(+) ATPase activity. H441 human airway epithelial cells were used to examine the effects of hypoxia on Na(+) transport, AMP : ATP ratio and AMPK activity. Lentiviral constructs were used to modify cellular AMPK abundance and activity; pharmacological agents were used to modify cellular ROS. AMPK was activated by exposure to 3% or 0.2% O(2) for 60 min in cells grown in submerged culture or when fluid (0.1 mL·cm(-2) ) was added to the apical surface of cells grown at the air-liquid interface. Only 0.2% O(2) activated AMPK in cells grown at the air-liquid interface. AMPK activation was associated with elevation of cellular AMP:ATP ratio and activity of the upstream kinase LKB1. Hypoxia inhibited basolateral ouabain-sensitive I(sc) (I(ouabain) ) and apical amiloride-sensitive Na(+) conductance (G(Na+) ). Modification of AMPK activity prevented the effect of hypoxia on I(ouabain) (Na(+) K(+) ATPase) but not apical G(Na+) . Scavenging of superoxide and inhibition of NADPH oxidase prevented the effect of hypoxia on apical G(Na+) (epithelial Na(+) channels). Hypoxia activates AMPK-dependent and -independent pathways in airway epithelial cells. Importantly, these pathways differentially regulate apical Na(+) channels and basolateral Na(+) K(+) ATPase activity to decrease transepithelial Na(+) transport. Luminal fluid potentiated the effect of hypoxia and activated AMPK, which could have important consequences in lung disease conditions. © 2012 The Authors. British Journal of Pharmacology © 2012 The British Pharmacological Society.

  13. AMP-activated protein kinase (AMPK)–dependent and –independent pathways regulate hypoxic inhibition of transepithelial Na+ transport across human airway epithelial cells

    PubMed Central

    Tan, CD; Smolenski, RT; Harhun, MI; Patel, HK; Ahmed, SG; Wanisch, K; Yáñez-Muñoz, RJ; Baines, DL

    2012-01-01

    BACKGROUND AND PURPOSE Pulmonary transepithelial Na+ transport is reduced by hypoxia, but in the airway the regulatory mechanisms remain unclear. We investigated the role of AMPK and ROS in the hypoxic regulation of apical amiloride-sensitive Na+ channels and basolateral Na+K+ ATPase activity. EXPERIMENTAL APPROACH H441 human airway epithelial cells were used to examine the effects of hypoxia on Na+ transport, AMP : ATP ratio and AMPK activity. Lentiviral constructs were used to modify cellular AMPK abundance and activity; pharmacological agents were used to modify cellular ROS. KEY RESULTS AMPK was activated by exposure to 3% or 0.2% O2 for 60 min in cells grown in submerged culture or when fluid (0.1 mL·cm−2) was added to the apical surface of cells grown at the air–liquid interface. Only 0.2% O2 activated AMPK in cells grown at the air–liquid interface. AMPK activation was associated with elevation of cellular AMP : ATP ratio and activity of the upstream kinase LKB1. Hypoxia inhibited basolateral ouabain-sensitive Isc (Iouabain) and apical amiloride-sensitive Na+ conductance (GNa+). Modification of AMPK activity prevented the effect of hypoxia on Iouabain (Na+K+ ATPase) but not apical GNa+. Scavenging of superoxide and inhibition of NADPH oxidase prevented the effect of hypoxia on apical GNa+ (epithelial Na+ channels). CONCLUSIONS AND IMPLICATIONS Hypoxia activates AMPK-dependent and -independent pathways in airway epithelial cells. Importantly, these pathways differentially regulate apical Na+ channels and basolateral Na+K+ ATPase activity to decrease transepithelial Na+ transport. Luminal fluid potentiated the effect of hypoxia and activated AMPK, which could have important consequences in lung disease conditions. PMID:22509822

  14. A residual granuloma in association with a dental implant.

    PubMed

    McCracken, Michael S; Chavali, Ramakiran V; Al-Naief, Nasser Said; Eleazer, Paul D

    2012-04-01

    At times, dental implants are placed into sites with a history of periapical pathology. Sometimes the infection is active, and other times the tooth may have been extracted years before implant placement. In either case, the possibility exists for long-term residual cysts or infections that can negatively impact the prognosis of the implant. In this case report, an implant is placed into a healed mandibular ridge several months after extraction of the tooth. A radiolucency was noted on routine radiographic examination 2 years later. Surgical inspection and histology revealed a periapical granuloma with acute and chronic inflammatory cells. After surgical curettage of the site, the patient healed without complication. Implants may develop apical pathology as a result of a preexisting long-term residual infection.

  15. Prevalence of different periapical lesions associated with human teeth and their correlation with the presence and extension of apical external root resorption.

    PubMed

    Vier, F V; Figueiredo, J A P

    2002-08-01

    The aim of this study was to determine the prevalence of various periapical pathologies and their association with the presence and extent of apical external inflammatory root resorption in human teeth. One hundred and four root apices from extracted teeth with periapical lesions were examined. Semi-serial sections of soft tissue lesions were stained with HE. The lesions were classified as noncystic or cystic, each with different degrees of acute inflammation: 0, 1, 2 and 3, increasing in severity. The root apices were analysed by SEM. External root resorption was classified according to site, as periforaminal or foraminal, and the extension of the resorbed area graded in increasing area as 0, 1, 2 or 3. Cysts accounted for 24.5% of the samples, 84% of which were associated with marked inflammation. The most prevalent diagnosis was noncystic periapical abscess with varying degrees of severity (63.7%). Periapical granuloma was not a frequent finding. SEM analysis showed that 42.2% of the root apices had periforaminal resorption extending over 50% of their circumference. When the foraminal resorption was evaluated, 28.7% had resorption affecting >50% of the periphery. Only 8.9% of the samples showed no periforaminal or foraminal resorption. In the sample of extracted teeth investigated, 24.5% of the periapical lesions were cysts. Most periapical lesions (84.3%) displayed acute inflammation, whether cystic or not. Periforaminal resorption was present in 87.3% of the cases, and foraminal resorption in 83.2%. Periforaminal and foraminal resorptions were independent entities. There was no association between external root resorption and the nature of the periapical lesions.

  16. The world of epithelial sheets.

    PubMed

    Honda, Hisao

    2017-06-01

    An epithelium is a layer of closely connected cells covering the body or lining a body cavity. In this review, several fundamental questions are addressed regarding the epithelium. (i) While an epithelium functions as barrier against the external environment, how is barrier function maintained during its construction? (ii) What determines the apical and basal sides of epithelial layer? (iii) Is there any relationship between the apical side of the epithelium and the apical membrane of an epithelial cell? (iv) Why are hepatocytes (liver cells) called epithelial, even though they differ completely from column-like shape of typical epithelial cells? Keeping these questions in mind, multiple shapes of epithelia were considered, extracting a few of their elemental processes, and constructing a virtual world of epithelia by combining them. Epithelial cells were also classified into several types based on the number of apical domains of each cell. In addition, an intracellular organelle was introduced within epithelial cells, the vacuolar apical compartment (VAC), which is produced within epithelial cells surrounded by external cell matrix (ECM). The VAC interacts with areas of cell-cell contact of the cell surface membrane and is converted to apical membrane. The properties of VACs enable us to answer the initial questions posed above. Finally, the genetic and molecular mechanisms of epithelial morphogenesis are discussed. © 2017 Japanese Society of Developmental Biologists.

  17. Apical extrusion of debris in primary molar root canals using mechanical and manual systems.

    PubMed

    Buldur, B; Hascizmeci, C; Aksoy, S; Nur Aydin, M; Guvendi, O N

    2018-03-01

    Apical extrusion of debris in primary root canal treatment has not been well elucidated. The purpose of this study is to compare the amount of apically extruded debris during the preparation of primary molar root canals using ProTaper, ProTaper Next, Self-adjusting File (SAF) and hand files. One hundred sixty extracted primary mandibular molar teeth were assigned to 2 groups: Group 1: Resorbed (n=80) and Group 2: Non-resorbed (n=80) and randomly to four subgroups (n=20 teeth for each subgroup) according to the instruments used, ProTaper, ProTaper Next, SAF, and hand file. The apically extruded debris was collected and dried in preweighed Eppendof tubes. The dry weight was calculated by subtracting the preoperative weight from the postoperative weight. Data were analysed statistically using the ANOVA and the Bonferroni post hoc t-test. The amount of apically extruded debris was significantly less for the non-resorbed group compared to the resorbed group (P<0.05). Regardless of the resorption groups, ProTaper Next and SAF extruded significantly less debris than did the ProTaper and hand files (P<0.05), while no statistically significant difference was found between ProTaper Next and SAF (P>0.05). All instruments caused apically extruded debris in primary teeth.

  18. Stability and function of a putative microtubule-organizing center in the human parasite Toxoplasma gondii

    PubMed Central

    Leung, Jacqueline M.; He, Yudou; Zhang, Fangliang; Hwang, Yu-Chen; Nagayasu, Eiji; Liu, Jun; Murray, John M.; Hu, Ke

    2017-01-01

    The organization of the microtubule cytoskeleton is dictated by microtubule nucleators or organizing centers. Toxoplasma gondii, an important human parasite, has an array of 22 regularly spaced cortical microtubules stemming from a hypothesized organizing center, the apical polar ring. Here we examine the functions of the apical polar ring by characterizing two of its components, KinesinA and APR1, and show that its putative role in templating can be separated from its mechanical stability. Parasites that lack both KinesinA and APR1 (ΔkinesinAΔapr1) are capable of generating 22 cortical microtubules. However, the apical polar ring is fragmented in live ΔkinesinAΔapr1 parasites and is undetectable by electron microscopy after detergent extraction. Disintegration of the apical polar ring results in the detachment of groups of microtubules from the apical end of the parasite. These structural defects are linked to a diminished ability of the parasite to move and invade host cells, as well as decreased secretion of effectors important for these processes. Together the findings demonstrate the importance of the structural integrity of the apical polar ring and the microtubule array in the Toxoplasma lytic cycle, which is responsible for massive tissue destruction in acute toxoplasmosis. PMID:28331073

  19. Ex vivo investigation on the postoperative integrity of the apical constriction after the sole use of electronic working length determination.

    PubMed

    Wolgin, Michael; Grundmann, Markus J; Tchorz, Jörg P; Frank, Wilhelm; Kielbassa, Andrej M

    2017-09-01

    The present study investigated the accuracy of root canal preparation with regard to the integrity of the apical constriction (AC) using two different working length determination approaches: (1) the electronic method of working length determination (EWLD), and (2) the radiologic "gold standard" method (GS). Simulation models were constructed by arranging extracted human teeth by means of silicon bolstered gingiva masks, along with a conductive medium (alginate). Electronic working length determination (group 1; EWLD) and radiologic plus initial electronic working length determination for posterior comparability (group 2; GS) preceded manual root canal preparation of teeth in both groups. Master cones were inserted according to working lengths obtained from the group specific method. Subsequently, root apices (n=36) were longitudinally sectioned using a diamond-coated bur. The distance between the achieved apical endpoint of the endodontic preparation and the apical constriction (AC) was measured using digital photography. Then, distances between radiologically identified apical endpoints and AC (GS-AC) were compared with the corresponding distances EWLD-AC. Moreover, the postoperative status of the AC was examined with regard to both preparation approaches. Differences between distances GS-AC and EWLD-AC were not statistically significant (p >0.401) (Mann-Whitney-U). Among EWLD samples, 83% of the master cones exhibiting tugback at final insertion terminated close to the apical constriction (±0.5 mm), and no impairment of the minor diameter's integrity was observed. The sole use of EWLD allowed for a high accuracy of measurements and granted precise preparation of the apical regions. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Anterior mandibular apical base augmentation in the surgical orthodontic treatment of mandibular retrusion.

    PubMed

    Brusati, R; Giannì, A B

    2005-12-01

    The authors describe a surgical technique alternative to traditional pre-surgical orthodontics in order to increase the apical base in mandibular retrusion (class II, division I). This subapical osteotomy, optimizing inferior incisal axis without dental extractions and a long orthodontic treatment, associated to genioplasty permits to obtain an ideal labio-dento-mental morphology. This procedure avoids in some cases the need of a mandibular advancement and, if necessary, it reduces his entity with obvious advantages.

  1. A posteriori registration and subtraction of periapical radiographs for the evaluation of external apical root resorption after orthodontic treatment.

    PubMed

    Kreich, Eliane Maria; Chibinski, Ana Cláudia; Coelho, Ulisses; Wambier, Letícia Stadler; Zedebski, Rosário de Arruda Moura; de Moraes, Mari Eli Leonelli; de Moraes, Luiz Cesar

    2016-03-01

    This study employed a posteriori registration and subtraction of radiographic images to quantify the apical root resorption in maxillary permanent central incisors after orthodontic treatment, and assessed whether the external apical root resorption (EARR) was related to a range of parameters involved in the treatment. A sample of 79 patients (mean age, 13.5±2.2 years) with no history of trauma or endodontic treatment of the maxillary permanent central incisors was selected. Periapical radiographs taken before and after orthodontic treatment were digitized and imported to the Regeemy software. Based on an analysis of the posttreatment radiographs, the length of the incisors was measured using Image J software. The mean EARR was described in pixels and relative root resorption (%). The patient's age and gender, tooth extraction, use of elastics, and treatment duration were evaluated to identify possible correlations with EARR. The mean EARR observed was 15.44±12.1 pixels (5.1% resorption). No differences in the mean EARR were observed according to patient characteristics (gender, age) or treatment parameters (use of elastics, treatment duration). The only parameter that influenced the mean EARR of a patient was the need for tooth extraction. A posteriori registration and subtraction of periapical radiographs was a suitable method to quantify EARR after orthodontic treatment, and the need for tooth extraction increased the extent of root resorption after orthodontic treatment.

  2. New propolis type from north-east Brazil: chemical composition, antioxidant activity and botanical origin.

    PubMed

    Ferreira, Joselena M; Fernandes-Silva, Caroline C; Salatino, Antonio; Negri, Giuseppina; Message, Dejair

    2017-08-01

    Propolis is a bee product with wide diversity of biological activity. It has a complex composition, which is dependent on its botanical source. The present study aimed to determine the chemical profile, antioxidant activity and botanical origin of two samples of a propolis type from two locations of the state of Rio Grande do Norte (RN, north-east Brazil). The standard chemical characteristics of the RN propolis are similar or superior to the internationally marketed Brazilian green propolis. RN propolis from two locations have high antioxidant activity, corresponding to 10% (municipality of Afonso Bezerra) and 13% (municipality of Alto do Rodrigues) of quercetin activity by the 2,2-diphenyl-1-picrylhydrazyl method and to 15% (both locations) by the β-carotene discoloration method. High-performance liquid chromatography with diode array detection (HPLC-DAD)-electrospray ionization-tandem mass spectrometry analyses revealed that most constituents of the RN propolis are flavonoids, mainly flavonols and chalcones. HPLC-DAD analysis of ethanol extracts revealed a great similarity between the chemical profile of RN propolis and shoot apices of 'jurema-preta' (Mimosa tenuiflora, Leguminosae, Mimosoideae). 'Jurema-preta' shoot apices are likely resin sources of RN propolis. The chemical characteristics and antioxidant property of RN propolis provide promising prospects for the introduction of this type of propolis into the apicultural market. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  3. Phase 1/2a Study of the Malaria Vaccine Candidate Apical Membrane Antigen-1 (AMA-1) Administered in Adjuvant System AS01B or AS02A

    DTIC Science & Technology

    2009-04-01

    purified saponin extract from the bark of the South American tree Quillaja saponaria, while AS02A is based on an oil-in-water emulsion with the same amounts... Technology in Health (PATH) Malaria Vaccine Initiative, Bethesda, Maryland, United States of America, 3 Department of Medical Microbiology, Radboud...Additionally, active immunization of rhesus monkeys with P. knowlesi AMA-1 adjuvanted in saponin resulted in some animals demonstrating a delayed

  4. Proliferative effects of apical, but not basal, matrix metalloproteinase-7 activity in polarized MDCK cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Harrell, Permila C.; McCawley, Lisa J.; Fingleton, Barbara

    2005-02-15

    Matrix metalloproteinase-7 (MMP-7) is primarily expressed in glandular epithelium. Therefore, its mechanism of action may be influenced by its regulated vectorial release to either the apical and/or basolateral compartments, where it would act on its various substrates. To gain a better understanding of where MMP-7 is released in polarized epithelium, we have analyzed its pattern of secretion in polarized MDCK cells expressing stably transfected human MMP-7 (MDCK-MMP-7), and HCA-7 and Caco2 human colon cancer cell lines. In all cell lines, latent MMP-7 was secreted to both cellular compartments, but was 1.5- to 3-fold more abundant in the basolateral compartment asmore » compared to the apical. However, studies in the MDCK system demonstrated that MMP-7 activity was 2-fold greater in the apical compartment of MDCK-MMP-7{sup HIGH}-polarized monolayers, which suggests the apical co-release of an MMP-7 activator. In functional assays, MMP-7 over-expression increased cell saturation density as a result of increased cell proliferation with no effect on apoptosis. Apical MMP-7 activity was shown to be responsible for the proliferative effect, which occurred, as demonstrated by media transfer experiments, through cleavage of an apical substrate and not through the generation of a soluble factor. Taken together, our findings demonstrate the importance of MMP-7 secretion in relation to its mechanism of action when expressed in a polarized epithelium.« less

  5. Larval body patterning and apical organs are conserved in animal evolution

    PubMed Central

    2014-01-01

    Background Planktonic ciliated larvae are characteristic for the life cycle of marine invertebrates. Their most prominent feature is the apical organ harboring sensory cells and neurons of largely undetermined function. An elucidation of the relationships between various forms of primary larvae and apical organs is key to understanding the evolution of animal life cycles. These relationships have remained enigmatic due to the scarcity of comparative molecular data. Results To compare apical organs and larval body patterning, we have studied regionalization of the episphere, the upper hemisphere of the trochophore larva of the marine annelid Platynereis dumerilii. We examined the spatial distribution of transcription factors and of Wnt signaling components previously implicated in anterior neural development. Pharmacological activation of Wnt signaling with Gsk3β antagonists abolishes expression of apical markers, consistent with a repressive role of Wnt signaling in the specification of apical tissue. We refer to this Wnt-sensitive, six3- and foxq2-expressing part of the episphere as the ‘apical plate’. We also unraveled a molecular signature of the apical organ - devoid of six3 but expressing foxj, irx, nkx3 and hox - that is shared with other marine phyla including cnidarians. Finally, we characterized the cell types that form part of the apical organ by profiling by image registration, which allows parallel expression profiling of multiple cells. Besides the hox-expressing apical tuft cells, this revealed the presence of putative light- and mechanosensory as well as multiple peptidergic cell types that we compared to apical organ cell types of other animal phyla. Conclusions The similar formation of a six3+, foxq2+ apical plate, sensitive to Wnt activity and with an apical tuft in its six3-free center, is most parsimoniously explained by evolutionary conservation. We propose that a simple apical organ - comprising an apical tuft and a basal plexus innervated by sensory-neurosecretory apical plate cells - was present in the last common ancestors of cnidarians and bilaterians. One of its ancient functions would have been the control of metamorphosis. Various types of apical plate cells would then have subsequently been added to the apical organ in the divergent bilaterian lineages. Our findings support an ancient and common origin of primary ciliated larvae. PMID:24476105

  6. Systemic antibiotics for symptomatic apical periodontitis and acute apical abscess in adults.

    PubMed

    Cope, Anwen; Francis, Nick; Wood, Fiona; Mann, Mala K; Chestnutt, Ivor G

    2014-06-26

    Dental pain can have a considerable detrimental effect on an individual's quality of life. Symptomatic apical periodontitis and acute apical abscess are common causes of dental pain and arise from an inflamed or necrotic dental pulp, or infection of the pulpless root canal system. Clinical guidelines recommend that the first-line treatment for teeth with symptomatic apical periodontitis or an acute apical abscess should be removal of the source of inflammation or infection by local, operative measures, and that systemic antibiotics are currently only recommended for situations where there is evidence of spreading infection (cellulitis, lymph node involvement, diffuse swelling) or systemic involvement (fever, malaise). Despite this, there is evidence that dentists continue to prescribe antibiotics for these conditions. There is concern that this could contribute to the development of antibiotic-resistant bacterial colonies both within the individual and within the community as a whole. To evaluate the effects of systemic antibiotics provided with or without surgical intervention (such as extraction, incision and drainage of a swelling or endodontic treatment), with or without analgesics, for symptomatic apical periodontitis or acute apical abscess in adults. We searched the following electronic databases: Cochrane Oral Health Group's Trials Register (to 1 October 2013); Cochrane Central Register of Controlled Trials (The Cochrane Library 2013, Issue 9); MEDLINE via OVID (1946 to 1 October 2013); EMBASE via OVID (1980 to 1 October 2013) and CINAHL via EBSCO (1980 to 1 October 2013). We searched the World Health Organization (WHO) International Trials Registry Platform and the US National Institutes of Health Trials Registry (ClinicalTrials.gov) on 1 October 2013 to identify ongoing trials. We searched for grey literature using OpenGrey (to 1 October 2013) and ZETOC Conference Proceedings (1993 to 1 October 2013). We placed no restrictions on the language or date of publication when searching the electronic databases. Randomised controlled trials of systemic antibiotics in adults with a clinical diagnosis of symptomatic apical periodontitis or acute apical abscess, with or without surgical intervention (considered in this situation to be extraction, incision and drainage or endodontic treatment) and with or without analgesics. Two review authors screened the results of the searches against inclusion criteria, extracted data and assessed risk of bias independently and in duplicate. We calculated mean differences (MD) (standardised mean difference (SMD) when different scales were reported) and 95% confidence intervals (CI) for continuous data and, where results were meta-analysed, we used a fixed-effect model as there were fewer than four studies. We contacted study authors to obtain missing information. We included two trials in this review, with 62 participants included in the analyses. Both trials were conducted in university dental schools in the USA and compared the effects of oral penicillin V potassium (penicillin VK) versus a matched placebo given in conjunction with a surgical intervention (total or partial pulpectomy) and analgesics to adults with acute apical abscess or symptomatic necrotic tooth (no signs of spreading infection or systemic involvement (fever, malaise)). We assessed one study as having a high risk of bias and the other study as having unclear risk of bias.The primary outcome variables presented were participant-reported pain and swelling (one trial also reported participant-reported percussion pain). One study reported the type and number of analgesics taken by participants. One study recorded the incidence of postoperative endodontic flare-ups (people who returned with symptoms that necessitated further treatment). Adverse effects as reported in one study were diarrhoea (one participant, placebo group) and fatigue and reduced energy postoperatively (one participant, antibiotic group). No studies reporting quality of life measurements were suitable for inclusion. Objective 1: systemic antibiotics versus placebo with surgical intervention and analgesics for symptomatic apical periodontitis or acute apical abscess. Two studies provided data for the comparison between systemic antibiotics (penicillin VK) and a matched placebo for adults with acute apical abscess or a symptomatic necrotic tooth. Participants in one study all underwent a total pulpectomy of the affected tooth while participants in the other study had their tooth treated by either partial or total pulpectomy. Participants in both trials received oral analgesics. There were no statistically significant differences in participant-reported measures of pain or swelling at any of the time points assessed within the review. The MD for pain (short ordinal numerical scale 0 to 3) was -0.03 (95% CI -0.53 to 0.47) at 24 hours; 0.32 (95% CI -0.22 to 0.86) at 48 hours and 0.08 (95% CI -0.38 to 0.54) at 72 hours. The SMD for swelling was 0.27 (95% CI -0.23 to 0.78) at 24 hours; 0.04 (95% CI -0.47 to 0.55) at 48 hours and 0.02 (95% CI -0.49 to 0.52) at 72 hours. The body of evidence was assessed as at very low quality. Objective 2: systemic antibiotics without surgical intervention for adults with symptomatic apical periodontitis or acute apical abscess. We found no studies that compared the effects of systemic antibiotics with a matched placebo delivered without a surgical intervention for symptomatic apical periodontitis or acute apical abscess in adults. There is very low quality evidence that is insufficient to determine the effects of systemic antibiotics on adults with symptomatic apical periodontitis or acute apical abscess.

  7. Calcium in Gravitropism of the Moss Pohlia nutans (Hedw.) Lindb. protonemata

    NASA Astrophysics Data System (ADS)

    Khorkavtsiv, O. Ya; Demkiv, O. T.; Khorkavtsiv, Ya. D.

    Protonemata of mosses of Pohlia nutans grow both by extension and division of single apical cells which are negatively gravitropic in darkness. The fluorescence of Ca2+ increased in the tip of apical cells from the first hours of gravitropism stimulation. Cytochemical investigations confirmed the existence of a well pronounced tip-to-base Ca2+-gradient, its formation being favoured by localization of calcium influx in the tip of the apical cell. Measurement of the cytochemical reaction intensity showed that the level of Ca2+-ATFase activity is low in apex and increases towards the base of the apical cell. The gravitropic protonemata and filaments which grew on the clinostat showed similar distributions of the Ca2+ and Ca2+-ATFase activity along the apical cell axis. Thus, these data demonstrate that growing apical cells of gravitropic protonemata have a high tip-to-base Ca2+ gradient, the Ca2+ transport being afforded by Ca2+-ATFase.

  8. The influence of irrigating solutions on the accuracy of the electronic apex locator facility in the Tri Auto ZX handpiece.

    PubMed

    Erdemir, A; Eldeniz, A U; Ari, H; Belli, S; Esener, T

    2007-05-01

    To determine the influence of various irrigating solutions on the accuracy of the electronic apex locator facility in the Tri Auto ZX handpiece. One hundred and forty teeth with single canals and mature apices, scheduled for extraction for either periodontal or prosthetic reasons in 76 patients were used. Following informed written consent local anaesthesia was administered, access cavities were prepared and pulp tissue removed. The teeth were then randomly divided into seven groups according to the irrigating solutions used. The root canal length measurements were completed using the Tri Auto ZX handpiece with automatic reverse function in the presence of one or other of the following solutions: 0.9% saline, 2.5% NaOCl, 3% H(2)O(2), 0.2% chlorhexidine, 17% EDTA, Ultracaine D-S or in the absence of an irrigating solution (control). Files were immobilized in the access cavity with composite resin. After extraction, the apical regions of the teeth were exposed and the file tips examined under a stereomicroscope. Distances between the file tips and the apical constriction were measured (mm) and analysed using a one-way anova and post hoc Tukey test. Mean distances from the apical constriction to the file tip were longer in the 0.9% saline group (P<0.05). There was no statistically significant difference on file tip position between the other solutions. Tri Auto ZX gave reliable results with all irrigating solutions apart from in the presence of 0.9% saline.

  9. [Measurement of periapical pressure created by occlusal loading].

    PubMed

    Dobó, Nagy Csaba; Fejérdy, Pál; Angyal, János; Harasztosi, Lajos; Daróczi, Lajos; Beke, Dezsó; Wesselink, Paul R

    2004-04-01

    The aim of this study was to develop an in vitro model in which the pressure in the periapical tissues can be measured during loading. Extracted human maxillary central incisors were embedded into resin blocks that had physical characteristics similar to bone and periodontal ligament. Each tooth was loaded with 20, 40, 50, 60, 75, 85, 100, 200, 300 and 450 N vertical forces from the incisal edge of the crown; this procedure was carried out three consecutive times. A minute resistor embedded in the periapical space was used to detect apical pressure changes during occlusal loading. The ratio of apical pressure changes (delta P) to the loading force changes (delta F) was calculated. The periapical pressure detected was in direct proportion to the loading force. The mean value of delta P/delta F was 5.994 kPa/N (SD = 2.04). Direct proportionality was found between the coronal loading and the apical hydrostatic pressure. The (delta P)/(delta F) ratio determined in this study makes it easier to estimate the apical hydrostatic pressure values during occlusal loading of single rooted teeth. In this study the apical pressure generated under occlusal loading was of the same magnitude as that estimated with the finite element method.

  10. Measurement of periapical pressure created by occlusal loading.

    PubMed

    Dobó-Nagy, C; Fejérdy, P; Angyal, J; Harasztosi, L; Daróczi, L; Beke, D; Wesselink, P R

    2003-10-01

    To develop an in vitro model in which the pressure in the periapical tissues can be measured during loading. Extracted human maxillary central incisors were embedded in resin blocks that had physical characteristics similar to those of bone and periodontal ligament. Each tooth was loaded with 20, 40, 50, 60, 75, 85, 100, 200, 300 and 450 N vertical forces from the incisal edge of the crown on three consecutive occasions. A minute resistor embedded in the periapical space was used to detect apical pressure changes during occlusal loading. The ratio of apical pressure changes (DeltaP) to the loading force changes (DeltaF) was calculated. The periapical pressure detected was in direct proportion to the loading force. The mean value of DeltaP/DeltaF was 5.994 kPa N-1 (SD = 2.04). Direct proportionality was found between the coronal loading and the apical hydrostatic pressure. The DeltaP/DeltaF ratio determined in this study makes it easier to estimate the apical hydrostatic pressure values during occlusal loading of single-rooted teeth. In this study, the apical pressure generated under occlusal loading was the same magnitude as that estimated with the finite element method.

  11. Palmatine, a protoberberine alkaloid, inhibits both Ca2+- and cAMP-activated Cl− secretion in isolated rat distal colon

    PubMed Central

    Wu, D Z; Yuan, J Y; Shi, H L; Hu, Z B

    2008-01-01

    Background and purpose: The protoberberine alkaloid berberine has been reported to inhibit colonic Cl− secretion. However, it is not known if other protoberberine alkaloids share these effects. We have therefore selected another protoberberine alkaloid, palmatine, to assess its effects on active ion transport across rat colonic epithelium. Experimental approach: Rat colonic mucosa was mounted in Ussing chambers and short circuit current (I SC), apical Cl− current and basolateral K+ current were recorded. Intracellular cAMP content was determined by an enzyme immunoassay. Intracellular Ca2+ concentration was measured with Fura-2 AM. Key results: Palmatine inhibited carbachol-induced Ca2+-activated Cl− secretion and the carbachol-induced increase of intracellular Ca2+ concentration. Palmatine also inhibited cAMP-activated Cl− secretion induced by prostaglandin E2 (PGE2) or forskolin. Palmatine prevented the elevation of intracellular cAMP by forskolin. Determination of apical Cl− currents showed that palmatine suppressed the forskolin-stimulated, apical cAMP-activated Cl− current but not the carbachol-stimulated apical Ca2+-activated Cl− current. Following permeabilization of apical membranes with nystatin, we found that palmatine inhibited a carbachol-stimulated basolateral K+ current that was sensitive to charybdotoxin and resistant to chromanol 293B. However, the forskolin-stimulated basolateral K+ current inhibited by palmatine was specifically blocked by chromanol 293B and not by charybdotoxin. Conclusions and implications: Palmatine attenuated Ca2+-activated Cl− secretion through inhibiting basolateral charybdotoxin-sensitive, SK4 K+ channels, whereas it inhibited cAMP-activated Cl− secretion by inhibiting apical CFTR Cl− channels and basolateral chromanol 293B-sensitive, KvLQT1 K+ channels. PMID:18204477

  12. Do Parameters Of Irradiation Influences The Apical Sealing Of Er:YAG Laser Apicetomies?

    NASA Astrophysics Data System (ADS)

    Marques, Aparecida Maria Cordeiro; dos Santos, Jean Nunes; Pinheiro, Antonio Luiz Barbosa

    2011-08-01

    Failures on the sealing of the tooth apex are responsible for many failures of apical surgeries. The Er:YAG laser has been proposed as an alternative for the use of rotator instruments on surgical endodontics. 12 human extracted canines had root endodontic treatment being the apical limit was set at 1mm before the apical foramen and were distributed into 2 groups. On group I, apicectomy was performed with the Er:YAG laser (250 mJ/15 Hz). Apical cut was performed of perpendicular mode with 3mm from the apical foramen. On Group II, the same procedures and the same sequence as above was used, varying only the parameters of the Er:YAG laser (400 mJ/6 Hz). The specimens were divided into groups and fixed, by the cervical third, on wax. Impermeabilization of the residual root apical third was performed following the same procedures used in the cervical third but the residual apex was left free from the impermeabilization. After that, the roots were immersed in a 2% methylene blue solution and placed in a bacteriological oven for 48 h. The segments were visually observed and the one showing greatest level of dye leakage was selected and kept in individual container and coded accordingly. Apical staining was measured using a stereoscopic magnifying glass; a compass; and caliper. The results showed that Group I showed significantly different higher mean level of dye leakage (5.67±4.9, p<0.05). There was a significantly difference between the groups. It is concluded that the apicectomies carried out with 400 mJ/6 Hz showed the smallest infiltration value.

  13. Expression of apical Na(+)-L-glutamine co-transport activity, B(0)-system neutral amino acid co-transporter (B(0)AT1) and angiotensin-converting enzyme 2 along the jejunal crypt-villus axis in young pigs fed a liquid formula.

    PubMed

    Yang, Chengbo; Yang, Xiaojian; Lackeyram, Dale; Rideout, Todd C; Wang, Zirong; Stoll, Barbara; Yin, Yulong; Burrin, Douglas G; Fan, Ming Z

    2016-06-01

    Gut apical amino acid (AA) transport activity is high at birth and during suckling, thus being essential to maintain luminal nutrient-dependent mucosal growth through providing AA as essential metabolic fuel, substrates and nutrient stimuli for cellular growth. Because system-B(0) Na(+)-neutral AA co-transporter (B(0)AT1, encoded by the SLC6A19 gene) plays a dominant role for apical uptake of large neutral AA including L-Gln, we hypothesized that high apical Na(+)-Gln co-transport activity, and B(0)AT1 (SLC6A19) in co-expression with angiotensin-converting enzyme 2 (ACE2) were expressed along the entire small intestinal crypt-villus axis in young animals via unique control mechanisms. Kinetics of Na(+)-Gln co-transport activity in the apical membrane vesicles, prepared from epithelial cells sequentially isolated along the jejunal crypt-villus axis from liquid formula-fed young pigs, were measured with the membrane potential being clamped to zero using thiocyanate. Apical maximal Na(+)-Gln co-transport activity was much higher (p < 0.05) in the upper villus cells than in the middle villus (by 29 %) and the crypt (by 30 %) cells, whereas Na(+)-Gln co-transport affinity was lower (p < 0.05) in the upper villus cells than in the middle villus and the crypt cells. The B(0)AT1 (SLC6A19) mRNA abundance was lower (p < 0.05) in the crypt (by 40-47 %) than in the villus cells. There were no significant differences in B(0)AT1 and ACE2 protein abundances on the apical membrane among the upper villus, the middle villus and the crypt cells. Our study suggests that piglet fast growth is associated with very high intestinal apical Na(+)-neutral AA uptake activities via abundantly co-expressing B(0)AT1 and ACE2 proteins in the apical membrane and by transcribing the B(0)AT1 (SLC6A19) gene in the epithelia along the entire crypt-villus axis.

  14. PGE(2) activation of apical membrane Cl(-) channels in A6 epithelia: impedance analysis.

    PubMed Central

    Păunescu, T G; Helman, S I

    2001-01-01

    Measurements of transepithelial electrical impedance of continuously short-circuited A6 epithelia were made at audio frequencies (0.244 Hz to 10.45 kHz) to investigate the time course and extent to which prostaglandin E(2) (PGE(2)) modulates Cl(-) transport and apical membrane capacitance in this cell-cultured model epithelium. Apical and basolateral membrane resistances were determined by nonlinear curve-fitting of the impedance vectors at relatively low frequencies (<50 Hz) to equations (Păunescu, T. G., and S. I. Helman. 2001. Biophys. J. 81:838--851) where depressed Nyquist impedance semicircles were characteristic of the membrane impedances under control Na(+)-transporting and amiloride-inhibited conditions. In all tissues (control, amiloride-blocked, and amiloride-blocked and furosemide-pretreated), PGE(2) caused relatively small (< approximately 3 microA/cm(2)) and rapid (<60 s) maximal increase of chloride current due to activation of a rather large increase of apical membrane conductance that preceded significant activation of Na(+) transport through amiloride-sensitive epithelial Na(+) channels (ENaCs). Apical membrane capacitance was frequency-dependent with a Cole-Cole dielectric dispersion whose relaxation frequency was near 150 Hz. Analysis of the time-dependent changes of the complex frequency-dependent equivalent capacitance of the cells at frequencies >1.5 kHz revealed that the mean 9.8% increase of capacitance caused by PGE(2) was not correlated in time with activation of chloride conductance, but rather correlated with activation of apical membrane Na(+) transport. PMID:11463630

  15. Evaluation of the Amount of Debris extruded apically by using Conv-entional Syringe, Endovac and Ultrasonic Irrigation Technique: An In Vitro Study

    PubMed Central

    Tambe, Varsha H; Nagmode, Pradnya S; Vishwas, Jayshree R; P, Saujanya K; Angadi, Prabakar; Ali, Fareedi Mukram

    2013-01-01

    Background: To compare the amount of debris extruded apically by using conventional syringe, Endovac & Ultrasonic irrigation. Materials & Methods: Thirty freshly extracted mandibular premolars were selected, working length was determined and mounted in a debris and collection apparatus. The canals were prepared. After each instrument change, 1 ml. of 3% sodium hypochlorite was used as irrigation. Debris extruded apically by using conventional syringe, endovac& ultrasonic irrigation tech, was measured using the electronic balance to determine its weight and statistical analysis was performed. The mean difference between the groups was determined using statistical analysis within the groups &between the groups for equal variances. Results: Among all the groups, significantly less debris were found apically in the Endovac group (0.96) compared to conventional and ultrasonic group (1.23) syringe. Conclusion: The present study showed that endovac system extrudes less amount of debris apically as compared to ultrasonic followed by conventional so incidence of flare up can be reduce by using endovac irrigation system. How to cite this article: Tambe V H, Nagmode P S, Vishwas J R, Saujanya K P, Angadi P, Ali F M. Evaluation of the Amount of Debris extruded apically by using Conventional Syringe, Endovac and Ultrasonic Irrigation Technique: An In Vitro Study. J Int Oral Health 2013; 5(3):63-66. PMID:24155604

  16. H(+)/solute-induced intracellular acidification leads to selective activation of apical Na(+)/H(+) exchange in human intestinal epithelial cells.

    PubMed

    Thwaites, D T; Ford, D; Glanville, M; Simmons, N L

    1999-09-01

    The intestinal absorption of many nutrients and drug molecules is mediated by ion-driven transport mechanisms in the intestinal enterocyte plasma membrane. Clearly, the establishment and maintenance of the driving forces - transepithelial ion gradients - are vital for maximum nutrient absorption. The purpose of this study was to determine the nature of intracellular pH (pH(i)) regulation in response to H(+)-coupled transport at the apical membrane of human intestinal epithelial Caco-2 cells. Using isoform-specific primers, mRNA transcripts of the Na(+)/H(+) exchangers NHE1, NHE2, and NHE3 were detected by RT-PCR, and identities were confirmed by sequencing. The functional profile of Na(+)/H(+) exchange was determined by a combination of pH(i), (22)Na(+) influx, and EIPA inhibition experiments. Functional NHE1 and NHE3 activities were identified at the basolateral and apical membranes, respectively. H(+)/solute-induced acidification (using glycylsarcosine or beta-alanine) led to Na(+)-dependent, EIPA-inhibitable pH(i) recovery or EIPA-inhibitable (22)Na(+) influx at the apical membrane only. Selective activation of apical (but not basolateral) Na(+)/H(+) exchange by H(+)/solute cotransport demonstrates that coordinated activity of H(+)/solute symport with apical Na(+)/H(+) exchange optimizes the efficient absorption of nutrients and Na(+), while maintaining pH(i) and the ion gradients involved in driving transport.

  17. Auditory brainstem activity and development evoked by apical versus basal cochlear implant electrode stimulation in children.

    PubMed

    Gordon, K A; Papsin, B C; Harrison, R V

    2007-08-01

    The role of apical versus basal cochlear implant electrode stimulation on central auditory development was examined. We hypothesized that, in children with early onset deafness, auditory development evoked by basal electrode stimulation would differ from that evoked more apically. Responses of the auditory nerve and brainstem, evoked by an apical and a basal implant electrode, were measured over the first year of cochlear implant use in 50 children with early onset severe to profound deafness who used hearing aids prior to implantation. Responses at initial stimulation were of larger amplitude and shorter latency when evoked by the apical electrode. No significant effects of residual hearing or age were found on initial response amplitudes or latencies. With implant use, responses evoked by both electrodes showed decreases in wave and interwave latencies reflecting decreased neural conduction time through the brainstem. Apical versus basal differences persisted with implant experience with one exception; eIII-eV interlatency differences decreased with implant use. Acute stimulation shows prolongation of basally versus apically evoked auditory nerve and brainstem responses in children with severe to profound deafness. Interwave latencies reflecting neural conduction along the caudal and rostral portions of the brainstem decreased over the first year of implant use. Differences in neural conduction times evoked by apical versus basal electrode stimulation persisted in the caudal but not rostral brainstem. Activity-dependent changes of the auditory brainstem occur in response to both apical and basal cochlear implant electrode stimulation.

  18. A posteriori registration and subtraction of periapical radiographs for the evaluation of external apical root resorption after orthodontic treatment

    PubMed Central

    Chibinski, Ana Cláudia; Coelho, Ulisses; Wambier, Letícia Stadler; Zedebski, Rosário de Arruda Moura; de Moraes, Mari Eli Leonelli; de Moraes, Luiz Cesar

    2016-01-01

    Purpose This study employed a posteriori registration and subtraction of radiographic images to quantify the apical root resorption in maxillary permanent central incisors after orthodontic treatment, and assessed whether the external apical root resorption (EARR) was related to a range of parameters involved in the treatment. Materials and Methods A sample of 79 patients (mean age, 13.5±2.2 years) with no history of trauma or endodontic treatment of the maxillary permanent central incisors was selected. Periapical radiographs taken before and after orthodontic treatment were digitized and imported to the Regeemy software. Based on an analysis of the posttreatment radiographs, the length of the incisors was measured using Image J software. The mean EARR was described in pixels and relative root resorption (%). The patient's age and gender, tooth extraction, use of elastics, and treatment duration were evaluated to identify possible correlations with EARR. Results The mean EARR observed was 15.44±12.1 pixels (5.1% resorption). No differences in the mean EARR were observed according to patient characteristics (gender, age) or treatment parameters (use of elastics, treatment duration). The only parameter that influenced the mean EARR of a patient was the need for tooth extraction. Conclusion A posteriori registration and subtraction of periapical radiographs was a suitable method to quantify EARR after orthodontic treatment, and the need for tooth extraction increased the extent of root resorption after orthodontic treatment. PMID:27051635

  19. Complex Apical Intraradicular Infection and Extraradicular Mineralized Biofilms as the Cause of Wet Canals and Treatment Failure: Report of 2 Cases.

    PubMed

    Ricucci, Domenico; Candeiro, George T M; Bugea, Calogero; Siqueira, José F

    2016-03-01

    This article describes 2 cases that showed persistent intracanal exudation (wet canal) even after several visits of antimicrobial endodontic treatment. Histologic and histobacteriologic investigation was conducted for determination of the cause. The 2 cases involved teeth with apical periodontitis lesions, which presented persistent exudation refractory to treatment after several visits. In case 1, it was not possible to achieve a dry canal, and surgery had to be performed. In case 2, attempts to dry the canal succeeded and the canal was filled, but follow-up examination showed an enlarged apical periodontitis lesion and extraction was performed. Biopsy specimens consisting of the root apex and apical periodontitis lesion for case 1 and the whole root for case 2 were subjected to histologic and histobacteriologic analyses. Both cases showed complex bacterial infection in the apical root, affecting both the intraradicular space and the outer root surface. Case 1 showed bacterial biofilms in ramifications, on untouched walls, and extending to the external root surface to form a thick and partially mineralized structure with high bacterial density. Different bacterial morphotypes were evidenced. Case 2 had a ledge on the apical canal wall created during instrumentation, which was filled with necrotic debris, filling material, and bacteria. The walls of the apical portion of the canal were covered by a bacterial biofilm, which was continuous with a thick extraradicular biofilm covering the cementum and dentin in resorptive defects. The extraradicular biofilm showed areas of mineralization and was dominated by filamentous bacteria. The 2 cases with wet canals and treatment failure were associated with complex persistent infection in the apical part of the root canal system extending to form thick and partially mineralized biofilm structures (calculus) on the outer apical root surface. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  20. Determining the apical terminus of root-end resected teeth using three modern apex locators: a comparative ex vivo study.

    PubMed

    ElAyouti, A; Kimionis, I; Chu, A-L; Löst, C

    2005-11-01

    To assess ex vivo the accuracy of various electronic apex locators in locating the apical terminus of root-end resected teeth. Ninety extracted human posterior teeth (182 root canals) were prepared to a minimum size of 40 and filled with gutta-percha and sealer. After resection of the apical 3 mm of the root, the root canal filling was removed using HERO rotary instruments. The size of the root canal at the apical terminus after removal of the filling ranged from size 50 to 90. The root canal length to the apical terminus was determined using 3 apex locators (Root ZX, Raypex4 and Apex Pointer). A new mounting model that utilized a micrometer was used to perform the measurements and to visually determine the actual position of the apical terminus. The frequency of locating the apical terminus and the corresponding 95% confidence interval (CI) were calculated. Additionally, the coefficient of repeatability of each apex locator and the limits of inter-operator agreement were determined. All apex locators showed an acceptable repeatability (0.02-0.03 mm coefficient of repeatability) and narrow limits of inter-operator agreement (+0.07 and -0.07 mm). The accuracy of determining the apical terminus within 1 mm in the root canal was as follows: Root ZX 90% (164/182 root-canals) [95%CI: 86-94%], Raypex4 74% (135/182 root-canals) [95%CI: 68-80%], and Apex Pointer 71% (129/182 root canals) [95%CI: 65-77%]. No over-instrumentation resulted when the Root ZX device was used. In contrast, using the Raypex4 or the Apex Pointer device resulted in over-instrumentation in 8 of 182 root canals (4%). Under the conditions of this study all three apex locators were able to detect the apical terminus of root-end resected teeth with an acceptable range. The Root ZX device was the most accurate without over-instrumentation of the root canals.

  1. Apical constriction is driven by a pulsatile apical myosin network in delaminating Drosophila neuroblasts.

    PubMed

    An, Yanru; Xue, Guosheng; Shaobo, Yang; Mingxi, Deng; Zhou, Xiaowei; Yu, Weichuan; Ishibashi, Toyotaka; Zhang, Lei; Yan, Yan

    2017-06-15

    Cell delamination is a conserved morphogenetic process important for the generation of cell diversity and maintenance of tissue homeostasis. Here, we used Drosophila embryonic neuroblasts as a model to study the apical constriction process during cell delamination. We observe dynamic myosin signals both around the cell adherens junctions and underneath the cell apical surface in the neuroectoderm. On the cell apical cortex, the nonjunctional myosin forms flows and pulses, which are termed medial myosin pulses. Quantitative differences in medial myosin pulse intensity and frequency are crucial to distinguish delaminating neuroblasts from their neighbors. Inhibition of medial myosin pulses blocks delamination. The fate of a neuroblast is set apart from that of its neighbors by Notch signaling-mediated lateral inhibition. When we inhibit Notch signaling activity in the embryo, we observe that small clusters of cells undergo apical constriction and display an abnormal apical myosin pattern. Together, these results demonstrate that a contractile actomyosin network across the apical cell surface is organized to drive apical constriction in delaminating neuroblasts. © 2017. Published by The Company of Biologists Ltd.

  2. Cementum as an age determinant: A forensic view.

    PubMed

    Raju, Godishala Swamy Sugunakar; Keerthi, Muddana; Nandan, Surapaneni Rateesh Kumar; Rao, Thokala Madhusudan; Kulkarni, Pavan G; Reddy, Dorankula Shyam Prasad

    2016-01-01

    Forensic age estimation (FAE) defines an expertise in forensic medicine, which aims to define in the most accurate way to determine the unknown chronological age of the person involved in judicial or legal proceedings. Dental cementum is a vital tissue which demonstrates continuous apposition throughout the life of the tooth. This appositional changes of cementum helps in approximation of age inforensic investigations. To correlate age by measuring the overlap or coronal migration of thecementum at thecementoenamel junction (CEJ) and the thickness of the cementum at the apical third of the root. A hundred freshly extracted teethfrom patients ranging from ages 17-55were longitudinal buccolingually ground sectioned using a mounted lathe wheel and Arkansas stone. 100 freshly extracted teeth of age group ranging from 17-55 years were taken. These teeth were longitudinally ground sectioned to a thickness of 8-10μm using a mounted lathe wheel and Arkansas stone. Afterwards the teeth were examined under a light microscope using a micrometer eyepiece for measuring the overlap or coronal migration of the cementum at the CEJ and the thickness of the cementum at the apical one-third of root. Measurements of the overlap or the coronal migration of the cementum at the CEJ and the thickness of the cementum at the apical one-third of the root are correlated with age. Results of the study indicated that the cementum at the CEJ migrated coronally during theaging process in case of the impacted teeth. There is also a significant increase in the thickness of the cementum at the apical onethird of rootin the case of both the impacted and erupted teeth. Approximation of age by measuring overlap or coronal migration of the cementum at the CEJ and the thickness of the cementum at the apical one-third of the rootsets new alleys in FAE.

  3. Marginal adaptation of newer root canal sealers to dentin: A SEM study.

    PubMed

    Polineni, Swapnika; Bolla, Nagesh; Mandava, Pragna; Vemuri, Sayesh; Mallela, Madhusudana; Gandham, Vijaya Madhuri

    2016-01-01

    This in vitro study evaluated and compared the marginal adaptation of three newer root canal sealers to root dentin. Thirty freshly extracted human single-rooted teeth with completely formed apices were taken. Teeth were decoronated, and root canals were instrumented. The specimens were randomly divided into three groups (n = 10) based upon the sealer used. Group 1 - teeth were obturated with epoxy resin sealer (MM-Seal). Group 2 - teeth were obturated with mineral trioxide aggregate (MTA) based sealer (MTA Fillapex), Group 3 - teeth were obturated with bioceramic sealer (EndoSequence BC sealer). Later samples were vertically sectioned using hard tissue microtome and marginal adaptation of sealers to root dentin was evaluated under coronal and apical halves using scanning electron microscopy (SEM) and marginal gap values were recorded. The data were statistically analyzed by two-way ANOVA and Tukey's multiple post hoc test. The highest marginal gap was seen in Group 2 (apical-16680.00 nm, coronal-10796 nm) and the lowest marginal gap was observed in Group 1 (apical-599.42 nm, coronal-522.72 nm). Coronal halves showed superior adaptation compared to apical halves in all the groups under SEM. Within the limitations of this study epoxy resin-based MM-Seal showed good marginal adaptation than other materials tested.

  4. Quantitative Analysis of the Area of the Apical Ectodermal Ridge in Chick Appendages Using Image-J.

    PubMed

    Syed, Hamd Binte Shahab; Khan, Muhammad Yunus

    2018-06-01

    To determine the effect of sodium phenytoin on the apical ectodermal ridges (AER) of chick wing buds by using the software program Image-J. An experimental study. Department of Anatomy, Regional Center, College of Physicians and Surgeons Pakistan (CPSP), Islamabad, from January 2014 to January 2015. Sixty fertilised chicken eggs of 'Egyptian fayoumi' breed were selected and separated into experimental (B) and control (A) groups, each having 30 eggs. A single dose of 3.5 mg sodium phenytoin was injected into each egg of the experimental group. The controls were injected with the same volume of normal saline. Developing embryos were extracted 96 hours (day 4) after incubation and histological sections were cut at 5 μm thickness. These sections were stained with Feulgen Nuclear and Light Green. The area of apical ectodermal ridges of chick wing buds was calculated by employing Image-J and subjected to statistical analysis. The difference between the mean values of the area of apical ectodermal ridges of experimental and control groups, as calculated by Image-J, was found to be statistically insignificant. Change in the area of the apical ectodermal ridges in experimental chicks, following phenytoin exposure, was insignificant as proven on the basis of quantification by Image-J.

  5. Micro-computed tomography study of the internal anatomy of mesial root canals of mandibular molars.

    PubMed

    Villas-Bôas, Marcelo Haas; Bernardineli, Norberti; Cavenago, Bruno Cavalini; Marciano, Marina; Del Carpio-Perochena, Aldo; de Moraes, Ivaldo Gomes; Duarte, Marco H; Bramante, Clovis M; Ordinola-Zapata, Ronald

    2011-12-01

    The aim of this study was to determine the mesiodistal and buccolingual diameter, apical volume, and the presence of isthmuses at the apical level of mesial root canals of mandibular molars. Sixty extracted first and second mandibular molars were scanned by using a SkyScan 1076 micro-computed tomography system with a voxel size of 18 μm. The apical thirds of the samples were reconstructed to allow a perpendicular section of the apical third by using the multiplanar reconstruction tool of the OsiriX software. The mesiodistal and the buccolingual distances of root canals were measured between the 1- to 4-mm levels. The type of root canal isthmuses present at these levels was classified by using modified criteria of Hsu and Kim. The volume of the root canal anatomy between the 1- to 3-mm apical levels was obtained by using the CTAN-CTVOL software. The medians of the mesiodistal diameter at the 1-, 2-, 3-, and 4-mm levels in the mesiobuccal and mesiolingual canals were 0.22 and 0.23 mm, 0.27 and 0.27 mm, 0.30 and 0.30 mm, and 0.36 and 0.35 mm, respectively. The buccolingual lengths at the 1-, 2-, 3-, and 4-mm levels were 0.37-0.35 mm, 0.55-0.41 mm, 0.54-0.49 mm, and 0.54 and 0.60 mm, respectively. The presence of isthmuses was more prevalent at the 3- to 4-mm level. However, 27 cases presented complete or incomplete isthmuses at the 1-mm apical level. The mean of the volume of the apical third was 0.83 mm(3), with a minimum value of 0.02 and a maximum value of 2.4 mm(3). Mesial root canals of mandibular molars do not present a consistent pattern. A high variability of apical diameters exists. The presence of isthmuses at the apical third was not uncommon even at the 1-mm apical level. Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  6. Pulp and apical tissue response to deep caries in immature teeth: A histologic and histobacteriologic study.

    PubMed

    Ricucci, Domenico; Siqueira, José F; Loghin, Simona; Lin, Louis M

    2017-01-01

    Descriptions of the pathologic changes in the pulp and associated apical structures of human immature teeth in response to deep caries are lacking in the literature. This article describes the histologic events associated with the radicular pulp and the apical tissues of human immature teeth following pulp inflammation and necrosis. Twelve immature teeth with destructive caries lesions were obtained from 8 patients. Two intact immature teeth served as controls. Teeth were extracted for reasons not related to this study and immediately processed for histopathologic and histobacteriologic analyses. Serial sections were examined for the pulp conditions and classified as reversible or irreversible pulp inflammation, or pulp necrosis. Other histologic parameters were also evaluated. In the 3 cases with reversible pulp inflammation, tissue in the pulp chamber showed mild to moderate inflammation and tertiary dentin formation related to tubules involved in the caries process. Overall, the radicular pulp tissue, apical papilla and Hertwig's epithelial root sheath (HERS) exhibited characteristics of normality. In the 3 cases with irreversible pulp inflammation, the pulps were exposed and severe inflammation occurred in the pulp chamber, with minor areas of necrosis and infection. Large areas of the canal walls were free from odontoblasts and lined by an atubular mineralized tissue. The apical papilla showed extremely reduced cellularity or lack of cells and HERS was discontinuous or absent. In the 6 cases with pulp necrosis, the coronal and radicular pulp tissue was necrotic and colonized by bacterial biofilms. The apical papilla could not be discerned, except for one case. HERS was absent in the necrotic cases. While immature teeth with reversible pulpitis showed histologic features almost similar to normal teeth in the canal and in the apical region, those with irreversible pulpitis and necrosis exhibited significant alterations not only in the radicular pulp but also in the apical tissues, including the apical papilla and HERS. Alterations in the radicular pulp and apical tissues help explain the outcome of current regenerative/reparative therapies and should be taken into account when devising more predictable therapeutic protocols for teeth with incomplete root formation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Cone-beam computed tomography evaluation of the association of cortical plate proximity and apical root resorption after orthodontic treatment.

    PubMed

    Nakada, Tomoo; Motoyoshi, Mitsuru; Horinuki, Eri; Shimizu, Noriyoshi

    2016-01-01

    We investigated the effects of proximity of the root apex to the maxillary labial cortical plate, palatal cortical plate, and incisive canal cortical plate on apical root resorption. Cone-beam computed tomography was used to measure the amount of root resorption and root apex movement around maxillary right and left central incisors in 30 adults who underwent four-bicuspid extraction followed by treatment with multibracket appliances. The patients were divided into three groups on the basis of the direction of root apex movement, after which the correlation between the amount of root resorption and root apex movement was determined. Mean apical root resorption was 1.80 ± 0.82 mm (range, 0.18-3.96 mm). The amount of root apex movement was positively correlated with the amount of root resorption on the side of pressure. Root apex proximity to the maxillary labial cortical plate, palatal cortical plate, and incisive canal cortical plate was associated with apical root resorption. Orthodontic treatment plans should carefully consider root proximity to the maxillary cortical plate. (J Oral Sci 58, 231-236, 2016).

  8. Detection of Apical Root Cracks Using Spectral Domain and Swept-source Optical Coherence Tomography.

    PubMed

    de Oliveira, Bruna Paloma; Câmara, Andréa Cruz; Duarte, Daniel Amancio; Gomes, Anderson Stevens Leonidas; Heck, Richard John; Antonino, Antonio Celso Dantas; Aguiar, Carlos Menezes

    2017-07-01

    This study aimed to evaluate the ability of 2 optical coherence tomographic (OCT) systems to detect apical dentinal microcracks. Twenty extracted human single-rooted mandibular incisors were selected. After root canal preparation with an R40 Reciproc file (VDW, Munich, Germany), the specimens presenting apical microcracks were identified using micro-computed tomographic (micro-CT) scanning as the gold standard. Then, the apical portions of the roots were imaged with spectral domain OCT (SD-OCT) and swept-source OCT systems, and the resulting images were blindly evaluated by 3 independent examiners to detect microcracks. The diagnostic performance of each OCT device was calculated, and statistical analysis was performed. Based on the micro-CT images, 12 (60%) roots presented dentinal microcracks in the apical region. The images generated by the OCT systems were able to show microcrack lines at the same location as the corresponding micro-CT cross sections. Although the diagnostic performance of the SD-OCT device was superior, there were no statistically significant differences between the 2 OCT devices (P > .05). Interexaminer agreement was substantial to almost perfect for the SD-OCT system and moderate to almost perfect for the swept-source OCT system, whereas intraexaminer agreement was substantial to almost perfect for both OCT devices. The detection ability verified for both OCT systems renders them promising tools for the diagnosis of apical microcracks. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  9. Polyphenols and phenolic acids from strawberry and apple decrease glucose uptake and transport by human intestinal Caco-2 cells.

    PubMed

    Manzano, Susana; Williamson, Gary

    2010-12-01

    The effect of polyphenols, phenolic acids and tannins (PPTs) from strawberry and apple on uptake and apical to basolateral transport of glucose was investigated using Caco-2 intestinal cell monolayers. Substantial inhibition on both uptake and transport was observed by extracts from both strawberry and apple. Using sodium-containing (glucose transporters SGLT1 and GLUT2 both active) and sodium-free (only GLUT2 active) conditions, we show that the inhibition of GLUT2 was greater than that of SGLT1. The extracts were analyzed and some of the constituent PPTs were also tested. Quercetin-3-O-rhamnoside (IC₅₀ =31 μM), phloridzin (IC₅₀=146 μM), and 5-caffeoylquinic acid (IC₅₀=2570 μM) contributed 26, 52 and 12%, respectively, to the inhibitory activity of the apple extract, whereas pelargonidin-3-O-glucoside (IC₅₀=802 μM) contributed 26% to the total inhibition by the strawberry extract. For the strawberry extract, the inhibition of transport was non-competitive based on kinetic analysis, whereas the inhibition of cellular uptake was a mixed-type inhibition, with changes in both V(max) and apparent K(m) . The results in this assay show that some PPTs inhibit glucose transport from the intestinal lumen into cells and also the GLUT2-facilitated exit on the basolateral side. Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Evaluation of Bacteriological Profile in the Apical Root Segment of the Patients with Primary Apical Periodontitis.

    PubMed

    Tatikonda, Aravind; Sudheep, N; Biswas, Krishna P; Gowtham, K; Pujari, Sudarshan; Singh, Padam

    2017-01-01

    Apical periodontitis usually results from bacterial accumulation and contamination occurring in the root-canal system, and extending beyond the apical foramen to involve the periapical tissues. Literature has a paucity of the studies that stress on the division and analysis of the pulp canal segments. The reason for this disparity might be the technique used for collecting the samples from the pulp canals. Hence, we carried out the present study to evaluate the microbial flora in the apical part of the roots with necrotic pulp canals. The present study included the assessment of 40 freshly extracted teeth that had necrotized pulpal tissue along with the presence of periapical periodontal lesions. Removal of the soft tissue lesions attached to the root portion of the teeth along with apical periodontal lesions was done with the help of scalpel blade, after rinsing them with a sterile solution of saline. Thorough cleaning of the root surfaces was done with hydrogen peroxide followed by rapid disinfection with the help of sodium hypochlorite at varying concentrations. Sectioning of the root portion of all the specimens with the help of a disk was done perpendicular to the long axis of the teeth at a distance of roughly 5 to 6 mm from the teeth's apicalmost point. Cryotubes were used for transferring the specimens of apical portions containing 1 mL of buffer and were subjected to immediate frozen processing at a temperature of -20°C. A 10 K-type file was used for the initial collection of the samples followed by subsequent incubation of the files and paper pints in the incubation cabinet. Subsequent deoxyribonucleic acid (DNA) extraction from the samples was done following the procedure described by Siqueira et al. Paster et al's modification of the reverse-capture checkerboard assay was used in the present study. Semiquantitative data were used for overcoming the difficulties arising due to obtaining the counts of the polymerase chain reaction (PCR)-based analysis of specimens. A positive result for the 16S ribosomal ribonucleic acid (rRNA) gene primer was observed only in two examined specimens of all the samples of the apical portion of the root canals in the present study. Negative result was shown by all the control group specimens, which were sterile samples. Presence of bacteria was confirmed by PCR in 38 out of 40 examined specimens. Amount of bacterial taxa, out of these 24 samples, ranged up to 6. Pseudoramibacter alactolyticus, Porphyromonas endodontalis, Dialister oral species, Bacteroidetes species, Streptococcus species, Olsenella uli, Synergistes species, Fusobacterium nucleatum, Parvimonas micra, Treponema denticola, and Filifactor alocis were the specific species detected. Bacteroidetes species was the only species that were detected at levels at or above 10 5 . Heavy bacterial infections were noticed in more than 45% of the cases at the periradicular part of the root canals. Microbial flora of the apical segment of the root with necrotized pulp tissue comprises a vast variety of pathogenic bacteria. For better prognosis of the treatment of such cases, adequate knowledge of the microbial flora of the root, especially the apical portion is necessary.

  11. Quantifying the extruded bacteria following use of two rotary instrumentation systems.

    PubMed

    Mohammadi, Zahed; Khademi, Abbasali

    2007-01-01

    All instrumentation techniques have been reported to be associated with extrusion of infected debris. The aim of this study was to evaluate the number of bacteria extruded apically from extracted teeth ex vivo after canal instrumentation using the two engine-driven techniques utilizing nickel-titanium instruments (Flex Master and Mtwo). Seventy extracted maxillary central incisor teeth were used. Access cavities were prepared and root canals were then contaminated with a suspension of Enterococcus faecalis and dried. The contaminated roots were divided into two experimental groups of 30 teeth each and one control group of 10 teeth. Group 1, Flex Master; Group2, Mtwo; Group 3, control group: no instrumentation was attempted. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples from the vials were incubated in culture media for 24 h. Colonies of bacteria were counted and the results were given as number of colony-forming units. The obtained data were analyzed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests, with α = 0.05 as the level for statistical significance. Findings showed that there was no significant difference as to the number of extruded bacteria between two engine-driven systems (P>0.05). Both engine-driven Nickel-Titanium systems extruded bacteria through the apical foramen.

  12. Effects of Cordyceps sinensis, Cordyceps militaris and their isolated compounds on ion transport in Calu-3 human airway epithelial cells.

    PubMed

    Yue, Grace Gar-Lee; Lau, Clara Bik-San; Fung, Kwok-Pui; Leung, Ping-Chung; Ko, Wing-Hung

    2008-04-17

    The traditional Chinese medicine Cordyceps sinensis (CS) (Clavicipitaceae) improves pulmonary function and is used to treat respiratory disease. Here, we compare the efficacy and mechanisms of action of Cordyceps sinensis and Cordyceps militaris (CM) (Clavicipitaceae) in Calu-3 human airway epithelial monolayer model. The extracts of Cordyceps sinensis and Cordyceps militaris, as well as their isolated compounds, cordycepin and adenosine, stimulated ion transport in a dose-dependent manner in Calu-3 monolayers. In subsequent experiments, transport inhibitor bumetanide and carbonic anhydrase inhibitor acetazolamide were added after Cordyceps sinensis and Cordyceps militaris extracts to determine their effects on Cl- and HCO3- movement. The results suggested that Cordyceps sinensis and Cordyceps militaris extracts may affect the anion movement from the basolateral to apical compartments in the airway epithelia. Basolateral Na+-K+-2Cl- cotransporter and apical cAMP-dependent cystic fibrosis transmembrane conductance regulator Cl- channel are involved in the process. The results provide the first evidence for the pharmacological mechanism of Cordyceps sinensis and Cordyceps militaris on respiratory tract.

  13. Influence of Apical Diameter on the Outcome of Regenerative Endodontic Treatment in Teeth with Pulp Necrosis: A Review.

    PubMed

    Fang, Yanjun; Wang, Xinhuan; Zhu, Jingjing; Su, Chaonan; Yang, Ying; Meng, Liuyan

    2018-03-01

    The aim of this review was to evaluate whether the apical diameter of teeth with necrotic pulp affects the outcomes of regenerative endodontic treatment and to determine the minimal apical size needed to obtain proper pulp revascularization. A literature search was performed from January 1, 2001, to November 25, 2016. Studies that satisfied the inclusion criteria were subjected to data extraction and analysis. In total, 14 studies with 85 patients were included. There were 10 case reports, 3 case series, and 1 prospective cohort study. The apical diameters of the teeth were divided into 3 groups: a narrow-sized group (group N), <0.5 mm (n = 10); a medium-sized group (group M), 0.5-1.0 mm (n = 25); and a wide-sized group (group W), >1.0 mm (n = 60). In group N, 1 tooth failed, 2 teeth completely healed, and 7 teeth incompletely healed. In group M, 2 teeth were excluded, and 1 tooth failed. In group W, 3 teeth were excluded, and 4 teeth failed. The clinical success rates were 90%, 95.65%, and 92.98% in groups N, M, and W, respectively. Within the limitations, the teeth with apical diameters <1.0 mm achieved clinical success after regenerative endodontic treatment. Meanwhile, the teeth with apical diameters of 0.5-1.0 mm attained the highest clinical success rate, which may be related to other potential factors, including patient age, pulp necrosis etiology, preoperative apical radiolucency, procedure details, follow-up period, and sample size. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  14. The effect of using an inverted master cone in a lateral compaction technique on the density of the gutta-percha fill.

    PubMed

    Wu, Min-Kai; de Groot, Sjoerd D; van der Sluis, Luc W M; Wesselink, Paul R

    2003-09-01

    We sought to measure and calculate the percentage of the gutta-percha-filled area in the apical root canal after the use of a standardized or inverted master cone in cold lateral compaction.Study design Two groups of extracted mandibular premolars with a single canal were instrumented with instruments of the same size; furthermore, they were obturated with laterally compacted gutta-percha cones with AH26 used as a sealer. In the first group, a standardized master cone was used with its narrow end in an apical position, whereas in the other group, an inverted master cone was used with its wide end in an apical position. The 2 master cones had the same apical diameter and fit in the apical canal. After lateral compaction, horizontal sections were cut at a level 3 and 5 mm from the apex of each filled tooth. Photographs of the sections were taken by using a microscope equipped with a digital camera; the photos were then scanned as tagged-image file format images. The cross-sectional area of the canal and the gutta-percha were measured by using an image-analysis program. The percentage of gutta-percha-filled area was calculated. At both levels, the inverted master cone produced a significantly higher percentage, statistically, of gutta-percha-filled area than did the standardized master cone (P =.001 at 3 mm; P =.012 at 5 mm). The use of an inverted master cone in cold lateral compaction may facilitate the apical placement of accessory cones, significantly increasing the volume of gutta-percha while reducing the volume of sealer in the apical root canal.

  15. Effect of the angle of apical resection on apical leakage, measured with a computerized fluid filtration device.

    PubMed

    Garip, Hasan; Garip, Yıldız; Oruçoğlu, Hasan; Hatipoğlu, Seda

    2011-03-01

    We determined the effect of the angle of apical resection on apical leakage using a computerized fluid filtration meter with a laser system and a digital air pressure regulator in 46 extracted single-rooted human teeth. Orthograde endodontic treatment was performed. The root canals were prepared up to a size 50 K-type file with 17% EDTA solution (Roth International, Chicago, IL) and 5% NaOCl solution as the irrigant. Gates Glidden burs (Maillefer Instruments, Ballaigues, Switzerland) were used to flare the coronal two thirds of the canal. All canals were dried with paper points and then obturated using cold lateral condensation (except for the positive controls) of gutta-percha points and AH plus (Dentsply DeTrey, Konstanz, Germany). All 40 roots were sectioned 3 mm from the apex. Forty teeth were assigned randomly into 1 of 4 experimental groups of 10 teeth each: in group 1, the teeth were resected apically (90° angle) and the cavities were obturated with mineral trioxide aggregate (MTA); in group 2, after apical resection (90° angle), a root-end cavity was prepared using ultrasonic diamond retrotips and the cavities were obturated with MTA; in group 3, the teeth were resected apically (∼45° angle) and the cavities were obturated with MTA; and in group 4, after apical resection (∼45° angle), a root-end cavity was prepared using ultrasonic diamond retrotips and the cavities were obturated with MTA. An additional 6 teeth were used as controls (3 each, negative and positive controls). Apical leakage was measured using a computerized fluid filtration meter with a laser system. The mean apical microleakage was 2.0 ± 0.4 × 10(-4), 1.6 ± 0.6 × 10(-4), 1.6 ± 0.9 × 10(-4), and 1.8 ± 0.7 × 10(-4) μL/cmH(2)O/min(-1) at 1.2 atm, in groups 1 to 4, respectively. Although the mean apical microleakage was greater in group 1, the differences among the 4 groups were not statistically significant (P > .05). The results of these in vitro studies showed that when an adequate retrograde cavity depth is prepared, variation in the root-end cutting angle does not necessarily cause any difference in microleakage. Copyright © 2011 Mosby, Inc. All rights reserved.

  16. In vitro uptake and immune functionality of digested Rosemary extract delivered through food grade vehicles.

    PubMed

    Arranz, E; Guri, A; Fornari, T; Mendiola, J A; Reglero, G; Corredig, M

    2017-07-01

    The digestion, absorption, uptake and bioavailability of a rosemary supercritical fluid extract encapsulated in oil in water emulsion were studied. Two emulsions with opposite surface charge were prepared, containing 7% canola oil, and either 2% lactoferrin or whey protein isolate. When absorption and uptake of carnosic acid and carnosol were followed on Caco-2 cell monolayers, there were no differences with protein type. However, when co-cultures of HT-29 MTX were employed, the presence of mucus caused a higher retention of carnosic acid in the apical layer for lactoferrin emulsions. The immune activity of the bioavailable fractions collected from cell absorption experiments was tested ex vivo on murine splenocytes. Although transport through the intestinal barrier models was low, the bioavailable fractions showed a significant effect on splenocytes proliferation. These results demonstrated the potential of using rosemary supercritical extract through protein stabilized oil in water emulsions, as a food with immunomodulatory functionality. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Light and nitrogen nutrition regulate apical control in Rosa hybrida L.

    PubMed

    Furet, Pierre-Maxime; Lothier, Jérémy; Demotes-Mainard, Sabine; Travier, Sandrine; Henry, Clémence; Guérin, Vincent; Vian, Alain

    2014-03-01

    Apical control is defined as the inhibition of basal axillary bud outgrowth by an upper actively growing axillary axis, whose regulation is poorly understood yet differs markedly from the better-known apical dominance. We studied the regulation of apical control by environmental factors in decapitated Rosa hybrida in order to remove the apical hormonal influence and nutrient sink. In this plant model, all the buds along the main axis have a similar morphology and are able to burst in vitro. We concentrated on the involvement of light intensity and nitrate nutrition on bud break and axillary bud elongation in the primary axis pruned above the fifth leaf of each rose bush. We observed that apical control took place in low light (92 μmol m(-2)s(-1)), where only the 2-apical buds grew out, both in low (0.25 mM) and high (12.25 mM) nitrate. In contrast, in high light (453 μmol m(-2)s(-1)), the apical control only operates in low nitrate while all the buds along the stem grew out when the plant was supplied with a high level of nitrate. We found a decreasing photosynthetic activity from the top to the base of the plant concomitant with a light gradient along the stem. The quantity of sucrose, fructose, glucose and starch are higher in high light conditions in leaves and stem. The expression of the sucrose transporter RhSUC2 was higher in internodes and buds in this lighting condition, suggesting an increased capacity for sucrose transport. We propose that light intensity and nitrogen availability both contribute to the establishment of apical control. Copyright © 2013 Elsevier GmbH. All rights reserved.

  18. Polarizing intestinal epithelial cells electrically through Ror2

    PubMed Central

    Cao, Lin; McCaig, Colin D.; Scott, Roderick H.; Zhao, Siwei; Milne, Gillian; Clevers, Hans; Zhao, Min; Pu, Jin

    2014-01-01

    ABSTRACT The apicobasal polarity of enterocytes determines where the brush border membrane (apical membrane) will form, but how this apical membrane faces the lumen is not well understood. The electrical signal across the epithelium could serve as a coordinating cue, orienting and polarizing enterocytes. Here, we show that applying a physiological electric field to intestinal epithelial cells, to mimic the natural electric field created by the transepithelial potential difference, polarized phosphorylation of the actin-binding protein ezrin, increased expression of intestinal alkaline phosphatase (ALPI, a differentiation marker) and remodeled the actin cytoskeleton selectively on the cathode side. In addition, an applied electric field also activated ERK1/2 and LKB1 (also known as STK11), key molecules in apical membrane formation. Disruption of the tyrosine protein kinase transmembrane receptor Ror2 suppressed activation of ERK1/2 and LKB1 significantly, and subsequently inhibited apical membrane formation in enterocytes. Our findings indicate that the endogenous electric field created by the transepithelial potential difference might act as an essential coordinating signal for apical membrane formation at a tissue level, through activation of LKB1 mediated by Ror2–ERK signaling. PMID:24928904

  19. Influence of the Apical Preparation Size and the Irrigant Type on Bacterial Reduction in Root Canal-treated Teeth with Apical Periodontitis.

    PubMed

    Rodrigues, Renata Costa Val; Zandi, Homan; Kristoffersen, Anne Karin; Enersen, Morten; Mdala, Ibrahimu; Ørstavik, Dag; Rôças, Isabela N; Siqueira, José F

    2017-07-01

    This clinical study evaluated the influence of the apical preparation size using nickel-titanium rotary instrumentation and the effect of a disinfectant on bacterial reduction in root canal-treated teeth with apical periodontitis. Forty-three teeth with posttreatment apical periodontitis were selected for retreatment. Teeth were randomly divided into 2 groups according to the irrigant used (2.5% sodium hypochlorite [NaOCl], n = 22; saline, n = 21). Canals were prepared with the Twisted File Adaptive (TFA) system (SybronEndo, Orange, CA). Bacteriological samples were taken before preparation (S1), after using the first instrument (S2), and then after the third instrument of the TFA system (S3). In the saline group, an additional sample was taken after final irrigation with 1% NaOCl (S4). DNA was extracted from the clinical samples and subjected to quantitative real-time polymerase chain reaction to evaluate the levels of total bacteria and streptococci. S1 from all teeth were positive for bacteria. Preparation to the first and third instruments from the TFA system showed a highly significant intracanal bacterial reduction regardless of the irrigant (P < .01). Apical enlargement to the third instrument caused a significantly higher decrease in bacterial counts than the first instrument (P < .01). Intergroup comparison revealed no significant difference between NaOCl and saline after the first instrument (P > .05). NaOCl was significantly better than saline after using the largest instrument in the series (P < .01). Irrespective of the type of irrigant, an increase in the apical preparation size significantly enhanced root canal disinfection. The disinfecting benefit of NaOCl over saline was significant at large apical preparation sizes. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  20. A laboratory comparison of apical leakage between immediate versus delayed post space preparation in root canals filled with Resilon.

    PubMed

    Attam, K; Talwar, S

    2010-09-01

    To analyse ex vivo the integrity of the apical seal of Resilon root fillings following immediate post space preparation or after 1 week when leaving either 5 mm or 3 mm of remaining filling material. One hundred and fifty freshly extracted mandibular premolar teeth with single, straight root canals were used. Teeth were autoclaved and cleaned of debris and soft tissue. After decoronation at 16 mm length, the root canals were instrumented using the K3 rotary system to apical file size 45, 0.04 taper with intermittent irrigation using 1% sodium hypochlorite. The teeth were randomly divided into five groups with 30 teeth in each group. Group 1: control group in which the root canals were not filled. Group 2: the canals were filled with Resilon and a post space prepared immediately leaving 5 mm of apical filling. Group 3: the canals were filled and post space preparation delayed for 1 week leaving 5 mm of material apically. Group 4: post space preparation in the canals was initiated immediately after filling leaving 3 mm of material apically. Group 5: the root canals were filled and post space preparation was delayed for 1 week leaving 3 mm of material apically. Microleakage was evaluated using a fluid transport device. The results were analysed statistically using one-way anova followed by Bonferroni's post hoc test. P < 0.05 was considered as statistically significant. There was a highly significant difference amongst the groups (P < 0.001). The maximum mean leakage was observed in group 5 (141.63 x 10(-6) microL min(-1) cm(-1)) and the least microleakage in group 2 (99.87 x 10(-6) microL min(-1) cm(-1)). Immediate post space preparation was associated with less microleakage than delayed preparation when both 5 mm and 3 mm of apical filling remained. Leaving 5 mm of Resilon provided a significantly better seal compared to leaving 3 mm of material.

  1. Chloride channel inhibition by a red wine extract and a synthetic small molecule prevents rotaviral secretory diarrhoea in neonatal mice

    PubMed Central

    Ko, Eun-A; Jin, Byung-Ju; Namkung, Wan; Ma, Tonghui; Thiagarajah, Jay R.; Verkman, A. S.

    2014-01-01

    Background Rotavirus is the most common cause of severe secretory diarrhoea in infants and young children globally. The rotaviral enterotoxin, NSP4, has been proposed to stimulate calcium-activated chloride channels (CaCC) on the apical plasma membrane of intestinal epithelial cells. We previously identified red wine and small molecule CaCC inhibitors. Objective To investigate the efficacy of a red wine extract and a synthetic small molecule, CaCCinh-A01, in inhibiting intestinal CaCCs and rotaviral diarrhoea. Design Inhibition of CaCC-dependent current was measured in T84 cells and mouse ileum. The effectiveness of an orally administered wine extract and CaCCinh-A01 in inhibiting diarrhoea in vivo was determined in a neonatal mouse model of rotaviral infection. Results Screening of ~150 red wines revealed a Cabernet Sauvignon that inhibited CaCC current in T84 cells with IC50 at a ~1:200 dilution, and higher concentrations producing 100% inhibition. A >1 kdalton wine extract prepared by dialysis, which retained full inhibition activity, blocked CaCC current in T84 cells and mouse intestine. In rotavirus-inoculated mice, oral administration of the wine extract prevented diarrhoea by inhibition of intestinal fluid secretion without affecting rotaviral infection. The wine extract did not inhibit the cystic fibrosis chloride channel (CFTR) in cell cultures, nor did it prevent watery stools in neonatal mice administered cholera toxin, which activates CFTR-dependent fluid secretion. CaCCinh-A01 also inhibited rotaviral diarrhoea. Conclusions Our results support a pathogenic role for enterocyte CaCCs in rotaviral diarrhoea and demonstrate the antidiarrhoeal action of CaCC inhibition by an alcohol-free, red wine extract and by a synthetic small molecule. PMID:24052273

  2. Neuroelectric Tuning of Cortical Oscillations by Apical Dendrites in Loop Circuits

    PubMed Central

    LaBerge, David; Kasevich, Ray S.

    2017-01-01

    Bundles of relatively long apical dendrites dominate the neurons that make up the thickness of the cerebral cortex. It is proposed that a major function of the apical dendrite is to produce sustained oscillations at a specific frequency that can serve as a common timing unit for the processing of information in circuits connected to that apical dendrite. Many layer 5 and 6 pyramidal neurons are connected to thalamic neurons in loop circuits. A model of the apical dendrites of these pyramidal neurons has been used to simulate the electric activity of the apical dendrite. The results of that simulation demonstrated that subthreshold electric pulses in these apical dendrites can be tuned to specific frequencies and also can be fine-tuned to narrow bandwidths of less than one Hertz (1 Hz). Synchronous pulse outputs from the circuit loops containing apical dendrites can tune subthreshold membrane oscillations of neurons they contact. When the pulse outputs are finely tuned, they function as a local “clock,” which enables the contacted neurons to synchronously communicate with each other. Thus, a shared tuning frequency can select neurons for membership in a circuit. Unlike layer 6 apical dendrites, layer 5 apical dendrites can produce burst firing in many of their neurons, which increases the amplitude of signals in the neurons they contact. This difference in amplitude of signals serves as basis of selecting a sub-circuit for specialized processing (e.g., sustained attention) within the typically larger layer 6-based circuit. After examining the sustaining of oscillations in loop circuits and the processing of spikes in network circuits, we propose that cortical functioning can be globally viewed as two systems: a loop system and a network system. The loop system oscillations influence the network system’s timing and amplitude of pulse signals, both of which can select circuits that are momentarily dominant in cortical activity. PMID:28659768

  3. Neuroelectric Tuning of Cortical Oscillations by Apical Dendrites in Loop Circuits.

    PubMed

    LaBerge, David; Kasevich, Ray S

    2017-01-01

    Bundles of relatively long apical dendrites dominate the neurons that make up the thickness of the cerebral cortex. It is proposed that a major function of the apical dendrite is to produce sustained oscillations at a specific frequency that can serve as a common timing unit for the processing of information in circuits connected to that apical dendrite. Many layer 5 and 6 pyramidal neurons are connected to thalamic neurons in loop circuits. A model of the apical dendrites of these pyramidal neurons has been used to simulate the electric activity of the apical dendrite. The results of that simulation demonstrated that subthreshold electric pulses in these apical dendrites can be tuned to specific frequencies and also can be fine-tuned to narrow bandwidths of less than one Hertz (1 Hz). Synchronous pulse outputs from the circuit loops containing apical dendrites can tune subthreshold membrane oscillations of neurons they contact. When the pulse outputs are finely tuned, they function as a local "clock," which enables the contacted neurons to synchronously communicate with each other. Thus, a shared tuning frequency can select neurons for membership in a circuit. Unlike layer 6 apical dendrites, layer 5 apical dendrites can produce burst firing in many of their neurons, which increases the amplitude of signals in the neurons they contact. This difference in amplitude of signals serves as basis of selecting a sub-circuit for specialized processing (e.g., sustained attention) within the typically larger layer 6-based circuit. After examining the sustaining of oscillations in loop circuits and the processing of spikes in network circuits, we propose that cortical functioning can be globally viewed as two systems: a loop system and a network system. The loop system oscillations influence the network system's timing and amplitude of pulse signals, both of which can select circuits that are momentarily dominant in cortical activity.

  4. P2 purinoceptors regulate calcium-activated chloride and fluid transport in 31EG4 mammary epithelia.

    PubMed

    Blaug, Sasha; Rymer, Jodi; Jalickee, Stephen; Miller, Sheldon S

    2003-04-01

    It has been reported that secretory mammary epithelial cells (MEC) release ATP, UTP, and UDP upon mechanical stimulation. Here we examined the physiological changes caused by ATP/UTP in nontransformed, clonal mouse mammary epithelia (31EG4 cells). In control conditions, transepithelial potential (apical side negative) and resistance were -4.4 +/- 1.3 mV (mean +/- SD, n = 12) and 517.7 +/- 39.4 Omega. cm(2), respectively. The apical membrane potential was -43.9 +/- 1.7 mV, and the ratio of apical to basolateral membrane resistance (R(A)/R(B)) was 3.5 +/- 0.2. Addition of ATP or UTP to the apical or basolateral membranes caused large voltage and resistance changes with an EC(50) of approximately 24 microM (apical) and approximately 30 microM (basal). Apical ATP/UTP (100 microM) depolarized apical membrane potential by 17.6 +/- 0.8 mV (n = 7) and decreased R(A)/R(B) by a factor of approximately 3. The addition of adenosine to either side (100 microM) had no effect on any of these parameters. The ATP/UTP responses were partially inhibited by DIDS and suramin and mediated by a transient increase in free intracellular Ca(2+) concentration (427 +/- 206 nM; 15-25 microM ATP, apical; n = 6). This Ca(2+) increase was blocked by cyclopiazonic acid, by BAPTA, or by xestospongin C. 31EG4 MEC monolayers also secreted or absorbed fluid in the resting state, and ATP or UTP increased fluid secretion by 5.6 +/- 3 microl x cm(-2) x h(-1) (n = 10). Pharmacology experiments indicate that 31EG4 epithelia contain P2Y(2) purinoceptors on the apical and basolateral membranes, which upon activation stimulate apical Ca(2+)-dependent Cl channels and cause fluid secretion across the monolayer. This suggests that extracellular nucleotides could play a fundamental role in mammary gland paracrine signaling and the regulation of milk composition in vivo.

  5. Isotropic actomyosin dynamics promote organization of the apical cell cortex in epithelial cells.

    PubMed

    Klingner, Christoph; Cherian, Anoop V; Fels, Johannes; Diesinger, Philipp M; Aufschnaiter, Roland; Maghelli, Nicola; Keil, Thomas; Beck, Gisela; Tolić-Nørrelykke, Iva M; Bathe, Mark; Wedlich-Soldner, Roland

    2014-10-13

    Although cortical actin plays an important role in cellular mechanics and morphogenesis, there is surprisingly little information on cortex organization at the apical surface of cells. In this paper, we characterize organization and dynamics of microvilli (MV) and a previously unappreciated actomyosin network at the apical surface of Madin-Darby canine kidney cells. In contrast to short and static MV in confluent cells, the apical surfaces of nonconfluent epithelial cells (ECs) form highly dynamic protrusions, which are often oriented along the plane of the membrane. These dynamic MV exhibit complex and spatially correlated reorganization, which is dependent on myosin II activity. Surprisingly, myosin II is organized into an extensive network of filaments spanning the entire apical membrane in nonconfluent ECs. Dynamic MV, myosin filaments, and their associated actin filaments form an interconnected, prestressed network. Interestingly, this network regulates lateral mobility of apical membrane probes such as integrins or epidermal growth factor receptors, suggesting that coordinated actomyosin dynamics contributes to apical cell membrane organization. © 2014 Klingner et al.

  6. Early extraction: a silver bullet to avoid nerve injury in lower third molar removal?

    PubMed

    Zhang, Q-B; Zhang, Z-Q

    2012-10-01

    This retrospective study evaluated the effects of early extraction of immature lower third molar on preventing complications, particularly nerve injury following lower third molar removal. Patients were grouped according to age and radiographic results: group A (518 patients, ≤23 years, immature teeth with apical foramen not closed); group B (532 patients, >23 years, mature teeth with closed apical foramen). Group A included 230 males and 288 females (average age 17 years). In group A, 808 lower mandibular third molars were extracted bilaterally in 290 and unilaterally in 228 patients; the incidence of complications was 2.48% (20/808) (all were temporary), the incidence of nerve injury was 0%. Group B included 250 males and 282 females (average age 39 years). In group B, 810 lower third molars were extracted bilaterally in 278 and unilaterally in 254 patients; the incidence of complications was 10% (81/810), the incidence of nerve injury was 1.6% (13/810). All complications were temporary, except two removals of permanent inferior alveolar nerve numbness (>6 months). In this study, early removal of the lower third molar was effective in avoiding some postoperative complications, especially nerve injury. Early extraction of lower third molar in youngsters is recommended following a team consultation. Copyright © 2012 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  7. Cytokinin is required for escape but not release from auxin mediated apical dominance

    PubMed Central

    Müller, Dörte; Waldie, Tanya; Miyawaki, Kaori; To, Jennifer PC; Melnyk, Charles W; Kieber, Joseph J; Kakimoto, Tatsuo; Leyser, Ottoline

    2015-01-01

    Auxin produced by an active primary shoot apex is transported down the main stem and inhibits the growth of the axillary buds below it, contributing to apical dominance. Here we use Arabidopsis thaliana cytokinin (CK) biosynthetic and signalling mutants to probe the role of CK in this process. It is well established that bud outgrowth is promoted by CK, and that CK synthesis is inhibited by auxin, leading to the hypothesis that release from apical dominance relies on an increased supply of CK to buds. Our data confirm that decapitation induces the expression of at least one ISOPENTENYLTRANSFERASE (IPT) CK biosynthetic gene in the stem. We further show that transcript abundance of a clade of the CK-responsive type-A Arabidopsis response regulator (ARR) genes increases in buds following CK supply, and that, contrary to their typical action as inhibitors of CK signalling, these genes are required for CK-mediated bud activation. However, analysis of the relevant arr and ipt multiple mutants demonstrates that defects in bud CK response do not affect auxin-mediated bud inhibition, and increased IPT transcript levels are not needed for bud release following decapitation. Instead, our data suggest that CK acts to overcome auxin-mediated bud inhibition, allowing buds to escape apical dominance under favourable conditions, such as high nitrate availability. Significance Statement It has been proposed that the release of buds from auxin-mediated apical dominance following decapitation requires increased cytokinin biosynthesis and consequent increases in cytokinin supply to buds. Here we show that in Arabidopsis, increases in cytokinin appear to be unnecessary for the release of buds from apical dominance, but rather allow buds to escape the inhibitory effect of apical auxin, thereby promoting bud activation in favourable growth conditions. PMID:25904120

  8. Healing of ungrafted and grafted extraction sockets after 12 weeks: a prospective clinical study.

    PubMed

    Heberer, Susanne; Al-Chawaf, Bassem; Jablonski, Carlo; Nelson, John J; Lage, Hermann; Nelson, Katje

    2011-01-01

    In this prospective study, bone formation in human extraction sockets augmented with Bio-Oss Collagen after a 12-week healing period was quantified and compared to bone formation in unaugmented extraction sockets. Selected patients with four-walled extraction sockets were included in this prospective study. After extraction, the sockets were randomly augmented using Bio-Oss Collagen or left to heal unfilled without raising a mucoperiosteal flap. At the time of implant placement, histologic specimens were obtained from the socket and analyzed. Statistical analysis was performed using the Wilcoxon signed-rank test. Twenty-five patients with a total of 39 sockets (20 augmented, 19 unaugmented) were included in the study and the histologic specimens analyzed. All specimens were free of inflammatory cells. The mean overall new bone formation in the augmented sites was 25% (range, 8%-41%) and in the unaugmented sockets it was 44% (range, 3%-79%). There was a significant difference in the rate of new bone formation between the grafted and ungrafted sockets and a significant difference in the bone formation rate in the apical compared to the coronal regions of all sockets, independent of the healing mode. This descriptive study demonstrated that bone formation in Bio-Oss Collagen-grafted human extraction sockets was lower than bone formation in ungrafted sockets. Bone formation occurred in all specimens with varying degrees of maturation independent of the grafting material and was initiated from the apical region.

  9. Effect of coronal flaring on apical extrusion of debris during root canal instrumentation using single-file systems.

    PubMed

    Topçuoğlu, H S; Üstün, Y; Akpek, F; Aktı, A; Topçuoğlu, G

    2016-09-01

    To evaluate the effect of coronal flaring on the amount of debris extruded apically during root canal preparation using the Reciproc, WaveOne (WO) and OneShape (OS) single-file systems. Ninety extracted single-rooted mandibular incisor teeth were randomly assigned to six groups (n = 15 for each group) for canal instrumentation. Endodontic access cavities were prepared in each tooth. In three of the six groups, coronal flaring was not performed; coronal flaring was performed with Gates-Glidden drills on all teeth in the remaining three groups. The canals were then instrumented with one or other of the following single-file instrument systems: Reciproc, WO and OS. Debris extruded apically during instrumentation was collected into pre-weighed Eppendorf tubes. The tubes were then stored in an incubator at 70 °C for 5 days. The weight of the dry extruded debris was established by subtracting the pre-instrumentation and post-instrumentation weight of the Eppendorf tubes for each group. Data were analysed using one-way analysis of variance (anova) and Tukey's post hoc tests (P = 0.05). Reciproc and WO files without coronal flaring produced significantly more debris compared with the other groups (P < 0.05). There was no significant difference in apical extrusion of debris amongst the other groups (P > 0.05). All single-file systems caused apical extrusion of debris. Performing coronal flaring prior to canal preparation reduced the amount of apically extruded debris when using Reciproc or WO systems. © 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  10. Comparative evaluation of apically extruded debris with V-Taper, ProTaper Next, and the Self-adjusting File systems.

    PubMed

    Vyavahare, Nishant K; Raghavendra, Srinidhi Surya; Desai, Niranjan N

    2016-01-01

    Complete cleaning of the root canal is the goal for ensuring success in endodontics. Removal of debris plays an important role in achieving this goal. In spite of advancements in instrument design, apical extrusion of debris remains a source of inflammation in the periradicular region. To comparatively evaluate the amount of apically extruded debris with V-Taper, ProTaper Next, and the self-adjusting File (SAF) system. Sixty-four extracted human mandibular teeth with straight root canals were taken. Access openings were done and working length determined. The samples were randomly divided into three groups: Group I - V-Taper files (n = 20), Group II - ProTaper Next (n = 20), Group III - SAF (n = 20). Biomechanical preparation was completed and the debris collected in vials to be quantitatively determined. The data obtained was statistically analyzed using ANOVA and post hoc Tukey's test. All the specimens showed apical debris extrusion. SAF showed significantly less debris extrusion compared to V-Taper and ProTaper Next (P < 0.001). Among Groups I and II, ProTaper Next showed lesser debris extrusion as compared to V-Taper, but it was not significant (P = 0.124). The SAF showed least amount of apical debris extrusion when compared to newer rotary endodontic instruments. This indicates that the incidence of inter-treatment flare-ups due to debris extrusion would be less with the SAF.

  11. Evaluation of apically extruded debris during root canal preparation in primary molar teeth using three different rotary systems and hand files.

    PubMed

    Topçuoğlu, Gamze; Topçuoğlu, Hüseyin Sinan; Akpek, Firdevs

    2016-09-01

    To assess the amount of debris extruded apically during root canal preparation using various nickel titanium instrumentation systems and hand files in primary molar teeth. Sixty extracted primary first mandibular molar human teeth were randomly assigned to four groups (n = 15 teeth for each group). The canals were then instrumented with the following instrument systems: Revo-S, Mtwo, ProTaper Next, and hand files. Apically extruded debris during instrumentation was collected into pre-weighed Eppendorf tubes. The Eppendorf tubes were then stored in an incubator at 70°C for 5 days. The weight of the dry extruded debris was established by subtracting the pre-instrumentation and post-instrumentation weight of the Eppendorf tubes for each group. The data were analyzed using one-way analysis of variance (anova) and Tukey's post hoc tests. ProTaper Next files were associated with less apically extruded debris than the Mtwo, Revo-S, and hand files (P < 0.05). Hand files extruded more debris than Mtwo and Revo-S instruments (P < 0.05), but there was no significant difference found between the Mtwo and Revo-S instruments (P > 0.05). All instruments were associated with apical extrusion of debris. ProTaper Next files caused less debris extrusion compared to the other systems used. © 2015 BSPD, IAPD and John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  12. Comparison of the amount of apical debris extrusion associated with different retreatment systems and supplementary file application during retreatment process.

    PubMed

    Çiçek, Ersan; Koçak, Mustafa Murat; Koçak, Sibel; Sağlam, Baran Can

    2016-01-01

    The type of instrument affects the amount of debris extruded. The aim of this study was to compare the effect of retreatment systems and supplementary file application on the amount of apical debris extrusion. Forty-eight extracted mandibular premolars with a single canal and similar length were selected. The root canals were prepared with the ProTaper Universal system with a torque-controlled engine. The root canals were dried and were obturated using Gutta-percha and sealer. The specimens were randomly divided into four equal groups according to the retreatment procedures (Group 1, Mtwo retreatment files; Group 2, Mtwo retreatment files + Mtwo rotary file #30 supplementary file; Group 3, ProTaper Universal retreatment (PTUR) files; and Group 4, PTUR files + ProTaper F3 supplementary file). The extruded debris during instrumentation were collected into preweighed Eppendorf tubes. The amount of apically extruded debris was calculated by subtracting the initial weight of the tube from the final weight. Three consecutive weights were obtained for each tube. No statistically significant difference was found in the amount of apically extruded debris between Groups 1 and 3 (P = 0.590). A significant difference was observed between Groups 1 and 2 (P < 0.05), and between Groups 3 and 4 (P < 0.05). The use of supplementary file significantly increased the amount of apically extruded debris.

  13. KIFC3, a microtubule minus end–directed motor for the apical transport of annexin XIIIb–associated Triton-insoluble membranes

    PubMed Central

    Noda, Yasuko; Okada, Yasushi; Saito, Nobuhito; Setou, Mitsutoshi; Xu, Ying; Zhang, Zheizeng; Hirokawa, Nobutaka

    2001-01-01

    We have identified and characterized a COOH-terminal motor domain–type kinesin superfamily protein (KIFC), KIFC3, in the kidney. KIFC3 is a minus end–directed microtubule motor protein, therefore it accumulates in regions where minus ends of microtubules assemble. In polarized epithelial cells, KIFC3 is localized on membrane organelles immediately beneath the apical plasma membrane of renal tubular epithelial cells in vivo and polarized MDCK II cells in vitro. Flotation assay, coupled with detergent extraction, demonstrated that KIFC3 is associated with Triton X-100–insoluble membrane organelles, and that it overlaps with apically transported TGN-derived vesicles. This was confirmed by immunoprecipitation and by GST pulldown experiments showing the specific colocalization of KIFC3 and annexin XIIIb, a previously characterized membrane protein for apically transported vesicles (Lafont, F., S. Lecat, P. Verkade, and K. Simons. 1998. J. Cell Biol. 142:1413–1427). Furthermore, we proved that the apical transport of both influenza hemagglutinin and annexin XIIIb was partially inhibited or accelerated by overexpression of motor-domainless (dominant negative) or full-length KIFC3, respectively. Absence of cytoplasmic dynein on these annexin XIIIb–associated vesicles and distinct distribution of the two motors on the EM level verified the existence of KIFC3-driven transport in epithelial cells. PMID:11581287

  14. Modulation of apical constriction by Wnt signaling is required for lung epithelial shape transition.

    PubMed

    Fumoto, Katsumi; Takigawa-Imamura, Hisako; Sumiyama, Kenta; Kaneiwa, Tomoyuki; Kikuchi, Akira

    2017-01-01

    In lung development, the apically constricted columnar epithelium forms numerous buds during the pseudoglandular stage. Subsequently, these epithelial cells change shape into the flat or cuboidal pneumocytes that form the air sacs during the canalicular and saccular (canalicular-saccular) stages, yet the impact of cell shape on tissue morphogenesis remains unclear. Here, we show that the expression of Wnt components is decreased in the canalicular-saccular stages, and that genetically constitutive activation of Wnt signaling impairs air sac formation by inducing apical constriction in the epithelium as seen in the pseudoglandular stage. Organ culture models also demonstrate that Wnt signaling induces apical constriction through apical actomyosin cytoskeletal organization. Mathematical modeling reveals that apical constriction induces bud formation and that loss of apical constriction is required for the formation of an air sac-like structure. We identify MAP/microtubule affinity-regulating kinase 1 (Mark1) as a downstream molecule of Wnt signaling and show that it is required for apical cytoskeletal organization and bud formation. These results suggest that Wnt signaling is required for bud formation by inducing apical constriction during the pseudoglandular stage, whereas loss of Wnt signaling is necessary for air sac formation in the canalicular-saccular stages. © 2017. Published by The Company of Biologists Ltd.

  15. The Balance of HCO3- Secretion vs. Reabsorption in the Endometrial Epithelium Regulates Uterine Fluid pH

    PubMed Central

    Xie, Zhang-Dong; Guo, Yi-Min; Ren, Mei-Juan; Yang, Jichun; Wang, Shao-Fang; Xu, Tong-Hui; Chen, Li-Ming; Liu, Ying

    2018-01-01

    Uterine fluid contains a high concentration of HCO3- which plays an essential role in sperm capacitation and fertilization. In addition, the HCO3- concentration in uterine fluid changes periodically during the estrous cycle. It is well-known that the endometrial epithelium contains machineries involving the apical SLC26 family anion exchangers for secreting HCO3- into the uterine fluid. In the present study, we find for the first time that the electroneutral Na+/HCO3- cotransporter NBCn1 is expressed at the apical membrane of the endometrial epithelium. The protein abundance of the apical NBCn1 and that of the apical SLC26A4 and SLC26A6 are reciprocally regulated during the estrous cycle in the uterus. NBCn1 is most abundant at diestrus, whereas SLC26A4/A6 are most abundant at proestrus/estrus. In the ovariectomized mice, the expression of uterine NBCn1 is inhibited by β-estradiol, but stimulated by progesterone, whereas that of uterine SLC26A4/A6 is stimulated by β-estradiol. In vivo perfusion studies show that the endometrial epithelium is capable of both secreting and reabsorbing HCO3-. Moreover, the activity for HCO3- secretion by the endometrial epithelium is significantly higher at estrus than it is at diestrus. The opposite is true for HCO3- reabsorption. We conclude that the endometrial epithelium simultaneously contains the activity for HCO3- secretion involving the apical SLC26A4/A6 and the activity for HCO3- reabsorption involving the apical NBCn1, and that the acid-base homeostasis in the uterine fluid is regulated by the finely-tuned balance of the two activities. PMID:29422866

  16. Characterization of tomato apical stunt viroid isolated from a 24-year old seed lot of Capsicum annuum.

    PubMed

    Verhoeven, J Th J; Koenraadt, H M S; Westenberg, M; Roenhorst, J W

    2017-06-01

    Tomato apical stunt viroid (TASVd) has been identified in a 24-year old seed lot of Capsicum annuum produced in Taiwan. It is the first finding of TASVd in this plant species. The isolate could be discriminated from all reported isolates of TASVd based on its nucleotide sequence, which showed only 94.8% identity with the most related genotype of TASVd. This discrimination was substantiated by phylogenetic analysis. Inoculation of a RNA extract of contaminated seeds to healthy pepper plants showed that the infectivity of the viroid had remained over time. Nevertheless, no transmission to seedlings was observed.

  17. Oxidative Stress in the Local and Systemic Events of Apical Periodontitis

    PubMed Central

    Hernández-Ríos, Patricia; Pussinen, Pirkko J.; Vernal, Rolando; Hernández, Marcela

    2017-01-01

    Oxidative stress is involved in the pathogenesis of a variety of inflammatory disorders. Apical periodontitis (AP) usually results in the formation of an osteolytic apical lesion (AL) caused by the immune response to endodontic infection. Reactive oxygen species (ROS) produced by phagocytic cells in response to bacterial challenge represent an important host defense mechanism, but disturbed redox balance results in tissue injury. This mini review focuses on the role of oxidative stress in the local and associated systemic events in chronic apical periodontitis. During endodontic infection, ligation of Toll-like receptors (TLRs) on phagocytes' surface triggers activation, phagocytosis, synthesis of ROS, activation of humoral and cellular responses, and production of inflammatory mediators, such as, cytokines and matrix metalloproteinases (MMPs). The increment in ROS perturbs the normal redox balance and shifts cells into a state of oxidative stress. ROS induce molecular damage and disturbed redox signaling, that result in the loss of bone homeostasis, increased pro-inflammatory mediators, and MMP overexpression and activation, leading to apical tissue breakdown. On the other hand, oxidative stress has been strongly involved in the pathogenesis of atherosclerosis, where a chronic inflammatory process develops in the arterial wall. Chronic AP is associated with an increased risk of cardiovascular diseases (CVD) and especially atherogenesis. The potential mechanisms linking these diseases are also discussed. PMID:29163211

  18. Role of the BK channel (KCa1.1) during activation of electrogenic K+ secretion in guinea pig distal colon

    PubMed Central

    Zhang, Jin; Halm, Susan T.

    2012-01-01

    Secretagogues acting at a variety of receptor types activate electrogenic K+ secretion in guinea pig distal colon, often accompanied by Cl− secretion. Distinct blockers of KCa1.1 (BK, Kcnma1), iberiotoxin (IbTx), and paxilline inhibited the negative short-circuit current (Isc) associated with K+ secretion. Mucosal addition of IbTx inhibited epinephrine-activated Isc (epiIsc) and transepithelial conductance (epiGt) consistent with K+ secretion occurring via apical membrane KCa1.1. The concentration dependence of IbTx inhibition of epiIsc yielded an IC50 of 193 nM, with a maximal inhibition of 51%. Similarly, IbTx inhibited epiGt with an IC50 of 220 nM and maximal inhibition of 48%. Mucosally added paxilline (10 μM) inhibited epiIsc and epiGt by ∼50%. IbTx and paxilline also inhibited Isc activated by mucosal ATP, supporting apical KCa1.1 as a requirement for this K+ secretagogue. Responses to IbTx and paxilline indicated that a component of K+ secretion occurred during activation of Cl− secretion by prostaglandin-E2 and cholinergic stimulation. Analysis of KCa1.1α mRNA expression in distal colonic epithelial cells indicated the presence of the ZERO splice variant and three splice variants for the COOH terminus. The presence of the regulatory β-subunits KCaβ1 and KCaβ4 also was demonstrated. Immunolocalization supported the presence of KCa1.1α in apical and basolateral membranes of surface and crypt cells. Together these results support a cellular mechanism for electrogenic K+ secretion involving apical membrane KCa1.1 during activation by several secretagogue types, but the observed K+ secretion likely required the activity of additional K+ channel types in the apical membrane. PMID:23064759

  19. Quantitative digital subtraction radiography in the assessment of external apical root resorption induced by orthodontic therapy: a retrospective study.

    PubMed

    Sunku, Raghavendra; Roopesh, R; Kancherla, Pavan; Perumalla, Kiran Kumar; Yudhistar, Palla Venkata; Reddy, V Sridhar

    2011-11-01

    The objective of this study was to evaluate density changes around the apices of teeth during orthodontic treatment by using digital subtraction radiography to measure the densities around six teeth (maxilla central incisors, lateral incisors, and canines) before and after orthodontic treatment in 36 patients and also assess treatment variables and their coorelation with root resorption. A total of 36 consecutive patient files were selected initially. The selected patients presented with a class I or II relationship and were treated with or without premolar extractions and fixed appliances. Some class II patients were treated additionally with extraoral forces or functional appliances. External apical root resorption (EARR) per tooth in millimeters was calculated and was also expressed as a percentage of the original root length. Image reconstruction and subtraction were performed using the software Regeemy Image Registration and Mosaicing (version 0.2.43-RCB, DPI-INPE, Sao Jose dos Campos, Sao Paulo, Brazil) by a single operator. A region of interest (ROI) was defined in the apical third of the root and density calibration was made in Image J® using enamel (gray value = 255) as reference in the same image. The mean gray values in the ROIs were reflective of the change in the density values between the two images. The root resorption of the tooth and the factors of malocclusion were analyzed with a one-way ANOVA. An independent t-test was performed to compare the mean amount of resorption between male and female, between extraction and nonextraction cases. The density changes after orthodontic treatment were analyzed using the Wilcoxon signedrank test. In addition, the density changes in different teeth were analyzed using the Kruskal-Wallis test. The cut-off for statistical significance was a p-value of 0.05. All the statistical analyses were carried out using SPSS (version 13.0 for Windows, Chicago, IL, USA). Gender, the age at which treatment was started and Angle's classification was not statistically related with observed root resorption. The mean percentage density reduction as assessed by DSR was greatest in both central incisor: by 27.2 and 25.2% in the upper-right and upper-left central incisors, respectively, followed by the upper-right and upper-left canine teeth (23.5 and 21.0%) and then the upper-right and upper-left lateral incisors (19.1 and 17.4%). Tooth extraction prior to treatment initiation and the duration of orthodontic treatment was positively correlated with the amount of root resorption. DSR is useful for evaluating density changes around teeth during orthodontic treatment. The density around the apices of teeth reduced significantly after the application of orthodontic forces during treatment. Assessment of density changes on treatment radiographs of patients undergoing orthodontic therapy may help in the monitoring of external apical root resorption during course of treatment.

  20. Visualization of removal of trapped air from the apical region of the straight root canal models generating 2-phase intermittent counter flow during ultrasonically activated irrigation.

    PubMed

    Peeters, Harry Huiz; Iskandar, Bernard; Suardita, Ketut; Suharto, Djoko

    2014-06-01

    The purpose of this in vitro study was to obtain a better understanding of the mechanism of irrigant traveling apically and generating 2-phase intermittent counter flow in straight root canal models during activation of the irrigant by ultrasonic means in an endodontic procedure. A high-speed imaging system, with high temporal and spatial resolution (FastCam SA5; Photron, Tokyo, Japan) at a frame rate of 100,000 frames per second using a macro lens (60 mm, f/2.8; Nikon, Tokyo, Japan), was used to visualize, in glass models of root canals, an ultrasonically induced acoustic pressure wave in an EDTA solution environment. A 25-mm stainless steel noncutting file #20 driven by an ultrasonic device (P5 Newtron; Satelec Acteon, Mérignac, France) at power settings of 5 and 7 produced disturbances at the solution-air interface. We found that apically directed travel of the irrigant was caused by disruption of the surface tension at the solution-air interface. This disruption caused by ultrasonic activation energy displaced air in the form of bubbles from the apical region toward the solution. The apical movement of the solution may be attributed to ultrasonically induced wave generation at the solution-air interface, resulting in the removal of trapped air from the root canal and allowing the solution to travel apically in the opposite directions (via a 2-phase intermittent counter flow). Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  1. High sodium intake increases HCO3− absorption in medullary thick ascending limb through adaptations in basolateral and apical Na+/H+ exchangers

    PubMed Central

    George, Thampi; Watts, Bruns A.

    2011-01-01

    A high sodium intake increases the capacity of the medullary thick ascending limb (MTAL) to absorb HCO3−. Here, we examined the role of the apical NHE3 and basolateral NHE1 Na+/H+ exchangers in this adaptation. MTALs from rats drinking H2O or 0.28 M NaCl for 5–7 days were perfused in vitro. High sodium intake increased HCO3− absorption rate by 60%. The increased HCO3− absorptive capacity was mediated by an increase in apical NHE3 activity. Inhibiting basolateral NHE1 with bath amiloride eliminated 60% of the adaptive increase in HCO3− absorption. Thus the majority of the increase in NHE3 activity was dependent on NHE1. A high sodium intake increased basolateral Na+/H+ exchange activity by 89% in association with an increase in NHE1 expression. High sodium intake increased apical Na+/H+ exchange activity by 30% under conditions in which basolateral Na+/H+ exchange was inhibited but did not change NHE3 abundance. These results suggest that high sodium intake increases HCO3− absorptive capacity in the MTAL through 1) an adaptive increase in basolateral NHE1 activity that results secondarily in an increase in apical NHE3 activity; and 2) an adaptive increase in NHE3 activity, independent of NHE1 activity. These studies support a role for NHE1 in the long-term regulation of renal tubule function and suggest that the regulatory interaction whereby NHE1 enhances the activity of NHE3 in the MTAL plays a role in the chronic regulation of HCO3− absorption. The adaptive increases in Na+/H+ exchange activity and HCO3− absorption in the MTAL may play a role in enabling the kidneys to regulate acid-base balance during changes in sodium and volume balance. PMID:21613418

  2. Immediate autotransplantation of mandibular third molar in China.

    PubMed

    Yan, Quanmei; Li, Bo; Long, Xing

    2010-10-01

    Tooth autotransplantation is a useful surgical method to replace a nonrestorable tooth. We reported our experiences in the replacement of mandibular nonrestorable molars by immediate autotransplantation in a Chinese population. Thirty-five mandibular third molars with open or closed apices from 34 patients were autotransplanted into the same or contralateral fresh recipient sites immediately after the extraction of the nonrestorable mandibular molars. Root canal treatment was routinely performed in the closed-apical molars within 1 month after surgery. Clinical and radiographic examination of the transplanted donor molars was done after surgery. Two teeth were been extracted for progressive root resorption. The remaining 33 autotransplanted teeth were asymptomatic and functioning after a mean follow-up period of 5.2 years. No infection, ankylosis, loss of the transplants, or root resorption was noted in the remaining autotransplanted teeth. Immediate autotransplantation of the mandibular third molar is a reasonable and alternative treatment to replace a nonrestorable tooth in China. Copyright © 2010 Mosby, Inc. All rights reserved.

  3. Frequency and levels of candidate endodontic pathogens in acute apical abscesses as compared to asymptomatic apical periodontitis.

    PubMed

    Rôças, Isabela N; Siqueira, José F

    2018-01-01

    Acute apical abscess is caused by bacteria that leave the infected dental root canal to invade the periodontal tissues. Most species occurring in abscesses are also found in asymptomatic infections; therefore, the possibility exists that not only the presence of certain species but also their specific counts influence the appearance of symptoms. This molecular study compared the frequency and levels of several candidate endodontic pathogens in teeth with acute apical abscesses and asymptomatic apical periodontitis. Samples were taken from the root canals of teeth with asymptomatic apical periodontitis (n = 73) and by aspiration of purulent exudate from acute abscesses (n = 55). DNA was extracted from samples and bacterial identifications were performed by a closed-ended semi-quantitative reverse-capture checkerboard approach targeting 40 bacterial species/phylotypes. Bacterial DNA was detected in all cases. In abscesses, the most prevalent taxa were Fusobacterium nucleatum (60%), Porphyromonas endodontalis (53%), Parvimonas micra (51%), and Streptococcus species (45%). The most frequently detected taxa in asymptomatic teeth were P. endodontalis (63%), Dialister invisus (58%), Olsenella uli (56%), and F. nucleatum (51%). None of the targeted taxa were significantly associated with abscesses when only mere presence was evaluated (p>0.05). However, semi-quantitative data demonstrated that P. endodontalis, Prevotella baroniae, Treponema denticola and Streptococcus species were significantly more frequent at levels >105 in abscesses than in asymptomatic cases (p<0.05). None of the target species/phylotypes were associated with abscesses in terms of frequency. However, some taxa were significantly found in higher levels in abscesses. Presence of a potentially virulent pathogen in high counts may increase the collective pathogenicity of the bacterial community and give rise to symptoms.

  4. Frequency and levels of candidate endodontic pathogens in acute apical abscesses as compared to asymptomatic apical periodontitis

    PubMed Central

    Rôças, Isabela N.

    2018-01-01

    Introduction Acute apical abscess is caused by bacteria that leave the infected dental root canal to invade the periodontal tissues. Most species occurring in abscesses are also found in asymptomatic infections; therefore, the possibility exists that not only the presence of certain species but also their specific counts influence the appearance of symptoms. This molecular study compared the frequency and levels of several candidate endodontic pathogens in teeth with acute apical abscesses and asymptomatic apical periodontitis. Methods Samples were taken from the root canals of teeth with asymptomatic apical periodontitis (n = 73) and by aspiration of purulent exudate from acute abscesses (n = 55). DNA was extracted from samples and bacterial identifications were performed by a closed-ended semi-quantitative reverse-capture checkerboard approach targeting 40 bacterial species/phylotypes. Results Bacterial DNA was detected in all cases. In abscesses, the most prevalent taxa were Fusobacterium nucleatum (60%), Porphyromonas endodontalis (53%), Parvimonas micra (51%), and Streptococcus species (45%). The most frequently detected taxa in asymptomatic teeth were P. endodontalis (63%), Dialister invisus (58%), Olsenella uli (56%), and F. nucleatum (51%). None of the targeted taxa were significantly associated with abscesses when only mere presence was evaluated (p>0.05). However, semi-quantitative data demonstrated that P. endodontalis, Prevotella baroniae, Treponema denticola and Streptococcus species were significantly more frequent at levels >105 in abscesses than in asymptomatic cases (p<0.05). Conclusion None of the target species/phylotypes were associated with abscesses in terms of frequency. However, some taxa were significantly found in higher levels in abscesses. Presence of a potentially virulent pathogen in high counts may increase the collective pathogenicity of the bacterial community and give rise to symptoms. PMID:29293651

  5. In Vitro Comparison of Apically Extruded Debris during Root Canal Preparation of Mandibular Premolars with Manual and Rotary Instruments.

    PubMed

    Soi, Sonal; Yadav, Suman; Sharma, Sumeet; Sharma, Mohit

    2015-01-01

    Background and aims. During root canal preparation, debris extruded beyond the apical foramen may result in periapical inflammation and postoperative pain. To date no root canal preparation method has been developed that extrudes no periapical debris. The purpose of this study was to identify a system leading to minimal extrusion of debris from the apical foramen. The study was conducted to comparatively evaluate the amount of apical extrusion of debris during root canal preparation using hand ProTaper and GT rotary and RaCe rotary instruments using crown-down technique. Materials and methods. Ninety freshly extracted human single-rooted mandibular premolars were equally assigned to three groups (n=30). The root canals were instrumented using hand ProTaper, GT rotary and RaCe rotary systems. Debris and irrigant extruded from the apical foramen were collected into vials. The mean weight of the remaining debris was calculated for each group and subjected to statistical analysis. Results. ANOVA was used to compare the mean dry weights of the debris extruded in the three groups, followedby post hoc Tukey tests for multiple comparisons the between groups. Highly significant differences were found in the amount of debris extruded among all the groups (P<0.001). The ProTaper group exhibited the highest mean debris weight (0.8293±0.05433 mg) and the RaCe system exhibited the lowest mean debris weight (0.1280±0.01606 mg). Conclusion. All the systems tested resulted in apical extrusion of debris. However, the hand ProTaper files extruded a significantly higher amount of debris than GT and RaCe systems.

  6. In Vitro Comparison of Apically Extruded Debris during Root Canal Preparation of Mandibular Premolars with Manual and Rotary Instruments

    PubMed Central

    Soi, Sonal; Yadav, Suman; Sharma, Sumeet; Sharma, Mohit

    2015-01-01

    Background and aims. During root canal preparation, debris extruded beyond the apical foramen may result in periapical inflammation and postoperative pain. To date no root canal preparation method has been developed that extrudes no periapical debris. The purpose of this study was to identify a system leading to minimal extrusion of debris from the apical foramen. The study was conducted to comparatively evaluate the amount of apical extrusion of debris during root canal preparation using hand ProTaper and GT rotary and RaCe rotary instruments using crown-down technique. Materials and methods. Ninety freshly extracted human single-rooted mandibular premolars were equally assigned to three groups (n=30). The root canals were instrumented using hand ProTaper, GT rotary and RaCe rotary systems. Debris and irrigant extruded from the apical foramen were collected into vials. The mean weight of the remaining debris was calculated for each group and subjected to statistical analysis. Results. ANOVA was used to compare the mean dry weights of the debris extruded in the three groups, followedby post hoc Tukey tests for multiple comparisons the between groups. Highly significant differences were found in the amount of debris extruded among all the groups (P<0.001). The ProTaper group exhibited the highest mean debris weight (0.8293±0.05433 mg) and the RaCe system exhibited the lowest mean debris weight (0.1280±0.01606 mg). Conclusion. All the systems tested resulted in apical extrusion of debris. However, the hand ProTaper files extruded a significantly higher amount of debris than GT and RaCe systems. PMID:26697144

  7. Five-year longitudinal assessment of the prognosis of apical microsurgery.

    PubMed

    von Arx, Thomas; Jensen, Simon S; Hänni, Stefan; Friedman, Shimon

    2012-05-01

    Apical surgery is an important treatment option for teeth with post-treatment apical periodontitis. Knowledge of the long-term prognosis is necessary when weighing apical surgery against alternative treatments. This study assessed the 5-year outcome of apical surgery and its predictors in a cohort for which the 1-year outcome was previously reported. Apical microsurgery procedures were uniformly performed using SuperEBA (Staident International, Staines, UK) or mineral trioxide aggregate (MTA) (ProRoot MTA; Dentsply Tulsa Dental Specialties, Tulsa, OK) root-end fillings or alternatively Retroplast capping (Retroplast Trading, Rorvig, Denmark). Subjects examined at 1 year (n = 191) were invited for the 5-year clinical and radiographic examination. Based on blinded, independent assessment by 3 calibrated examiners, the dichotomous outcome (healed or nonhealed) was determined and associated with patient-, tooth-, and treatment-related variables using logistic regression. At the 5-year follow-up, 9 of 191 teeth were unavailable, 12 of 191 teeth were extracted, and 170 of 191 teeth were examined (87.6% recall rate). A total of 129 of 170 teeth were healed (75.9%) compared with 83.8% at 1 year, and 85.3% were asymptomatic. Two significant outcome predictors were identified: the mesial-distal bone level at ≤ 3 mm versus >3 mm from the cementoenamel junction (78.2% vs 52.9% healed, respectively; odds ratio = 5.10; confidence interval, 1.67-16.21; P < .02) and root-end fillings with ProRoot MTA versus SuperEBA (86.4% vs. 67.3% healed, respectively; odds ratio = 7.65; confidence interval, 2.60-25.27; P < .004). This study suggested that the 5-year prognosis after apical microsurgery was 8% poorer than assessed at 1 year. It also suggested that the prognosis was significantly impacted by the interproximal bone levels at the treated tooth and by the type of root-end filling material used. Copyright © 2012 American Association of Endodontists. All rights reserved.

  8. Comparison of Apical Transportation with the Use of Rotary System and Reciprocating Handpiece with Precurved Hand Files: An In Vitro Study

    PubMed Central

    Mohammadian, Fatemeh; Sadeghi, Atefeh; Dibaji, Fatemeh; Sadegh, Mona; Ghoncheh, Zahra; Kharrazifard, Mohammad Javad

    2017-01-01

    Introduction: Success of root canal treatment depends on several factors; among which, maintaining the original canal path during mechanical preparation is extremely important. This in vitro study aimed to compare apical transportation using RaCe NiTi rotary system and precurved stainless steel (SS) hand files in a reciprocating handpiece. Methods and Materials: Mesiobuccal canals of 40 extracted human mandibular first and second molars with 20 to 45° curvatures and 3 to 7 mm curve radius were chosen for this study. After working length determination, the teeth were divided into two groups (n=20). Root canals were prepared with RaCe in group 1 and NSK handpiece and precurved SS hand files in group 2 up to #30 with 2% taper in both groups. Radiographs were taken of teeth before and after instrumentation from buccolingual and mesiodistal directions. The images were superimposed using Adobe Photoshop CS3 software. Degree of straightening and amount of apical transportation at 0, 0.5, 1, 2, 3, 4 and 5 mm levels short of the working length were determined using digital subtraction radiography. The student’s t test was used to compare the degree of straightening and Mann Whitney test was applied to compare apical transportation (millimeters) between the two groups. Results: No significant difference was noted between the two groups on buccolingual or mesiodistal views in degree of straightening and apical transportation on buccolingual view (P>0.05). However, on mesiodistal view, NSK reciprocating handpiece caused greater apical transportation at 0. 0.5 and 1 mm levels (P<0.05). Conclusion: The RaCe system and precurved SS files in reciprocating handpiece were highly similar in terms of degree of straightening and apical transportation. Thus, engine-driven NSK reciprocating handpiece can be used as an efficient adjunct for root canal preparation. PMID:29225642

  9. Comparison of Apical Transportation with the Use of Rotary System and Reciprocating Handpiece with Precurved Hand Files: An In Vitro Study.

    PubMed

    Mohammadian, Fatemeh; Sadeghi, Atefeh; Dibaji, Fatemeh; Sadegh, Mona; Ghoncheh, Zahra; Kharrazifard, Mohammad Javad

    2017-01-01

    Success of root canal treatment depends on several factors; among which, maintaining the original canal path during mechanical preparation is extremely important. This in vitro study aimed to compare apical transportation using RaCe NiTi rotary system and precurved stainless steel (SS) hand files in a reciprocating handpiece. Mesiobuccal canals of 40 extracted human mandibular first and second molars with 20 to 45 ° curvatures and 3 to 7 mm curve radius were chosen for this study. After working length determination, the teeth were divided into two groups ( n =20). Root canals were prepared with RaCe in group 1 and NSK handpiece and precurved SS hand files in group 2 up to #30 with 2% taper in both groups. Radiographs were taken of teeth before and after instrumentation from buccolingual and mesiodistal directions. The images were superimposed using Adobe Photoshop CS3 software. Degree of straightening and amount of apical transportation at 0, 0.5, 1, 2, 3, 4 and 5 mm levels short of the working length were determined using digital subtraction radiography. The student's t test was used to compare the degree of straightening and Mann Whitney test was applied to compare apical transportation (millimeters) between the two groups. No significant difference was noted between the two groups on buccolingual or mesiodistal views in degree of straightening and apical transportation on buccolingual view ( P >0.05). However, on mesiodistal view, NSK reciprocating handpiece caused greater apical transportation at 0. 0.5 and 1 mm levels ( P <0.05) . The RaCe system and precurved SS files in reciprocating handpiece were highly similar in terms of degree of straightening and apical transportation. Thus, engine-driven NSK reciprocating handpiece can be used as an efficient adjunct for root canal preparation.

  10. Actomyosin-based tissue folding requires a multicellular myosin gradient

    PubMed Central

    Miller, Pearson W.; Chanet, Soline; Stoop, Norbert; Dunkel, Jörn

    2017-01-01

    Tissue folding promotes three-dimensional (3D) form during development. In many cases, folding is associated with myosin accumulation at the apical surface of epithelial cells, as seen in the vertebrate neural tube and the Drosophila ventral furrow. This type of folding is characterized by constriction of apical cell surfaces, and the resulting cell shape change is thought to cause tissue folding. Here, we use quantitative microscopy to measure the pattern of transcription, signaling, myosin activation and cell shape in the Drosophila mesoderm. We found that cells within the ventral domain accumulate different amounts of active apical non-muscle myosin 2 depending on the distance from the ventral midline. This gradient in active myosin depends on a newly quantified gradient in upstream signaling proteins. A 3D continuum model of the embryo with induced contractility demonstrates that contractility gradients, but not contractility per se, promote changes to surface curvature and folding. As predicted by the model, experimental broadening of the myosin domain in vivo disrupts tissue curvature where myosin is uniform. Our data argue that apical contractility gradients are important for tissue folding. PMID:28432215

  11. In vitro comparison in a manikin model: increasing apical enlargement with K3 and K3XF rotary instruments.

    PubMed

    Olivieri, Juan Gonzalo; Stöber, Eva; García Font, Marc; González, Jose Antonio; Bragado, Pablo; Roig, Miguel; Duran-Sindreu, Fernando

    2014-09-01

    The aim of the study was to compare the K3 and K3XF systems (SybronEndo, Glendora, CA) after 1 and 2 uses by evaluating apical transportation, working length loss, and working time in a manikin model. Mesial canals of 40 extracted first mandibular molars were instrumented. Radiographs taken after instrumentation with #25, #30, #35, and #40 files were superimposed on the preoperative image in both mesiodistal and buccolingual angulations. AutoCAD (Autodesk Inc, San Rafael, CA) was used to measure working length loss and apical transportation at 0, 0.5, and 1 mm from the working length (WL). The working time was measured. Group comparison was analyzed using post hoc Tukey honestly significant difference tests (P < .05). No significant differences were found in apical transportation, working length loss between K3 and K3XF systems, or between the number of uses. Significant differences were found when canal enlargement was performed to a #35-40 (P < .05). K3 instrumentation performed significantly faster (29.6 ± 15.4) than with the K3XF system (40.2 ± 17.7) (P < .05). No differences were observed in working time when comparing the number of uses. K3 and R-phase K3XF rotary systems shaped curved root canals safely with minimal apical transportation, even up to a 40/04 file. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  12. Comparative evaluation of apically extruded debris with V-Taper, ProTaper Next, and the Self-adjusting File systems

    PubMed Central

    Vyavahare, Nishant K.; Raghavendra, Srinidhi Surya; Desai, Niranjan N.

    2016-01-01

    Background: Complete cleaning of the root canal is the goal for ensuring success in endodontics. Removal of debris plays an important role in achieving this goal. In spite of advancements in instrument design, apical extrusion of debris remains a source of inflammation in the periradicular region. Aim: To comparatively evaluate the amount of apically extruded debris with V-Taper, ProTaper Next, and the self-adjusting File (SAF) system. Materials and Methods: Sixty-four extracted human mandibular teeth with straight root canals were taken. Access openings were done and working length determined. The samples were randomly divided into three groups: Group I - V-Taper files (n = 20), Group II - ProTaper Next (n = 20), Group III - SAF (n = 20). Biomechanical preparation was completed and the debris collected in vials to be quantitatively determined. The data obtained was statistically analyzed using ANOVA and post hoc Tukey's test. Results: All the specimens showed apical debris extrusion. SAF showed significantly less debris extrusion compared to V-Taper and ProTaper Next (P < 0.001). Among Groups I and II, ProTaper Next showed lesser debris extrusion as compared to V-Taper, but it was not significant (P = 0.124). Conclusion: The SAF showed least amount of apical debris extrusion when compared to newer rotary endodontic instruments. This indicates that the incidence of inter-treatment flare-ups due to debris extrusion would be less with the SAF. PMID:27217636

  13. Molecular characterization of the microbial flora residing at the apical portion of infected root canals of human teeth.

    PubMed

    Chugal, Nadia; Wang, Jen-Kuei; Wang, Renke; He, Xuesong; Kang, Mo; Li, Jiyao; Zhou, Xuedong; Shi, Wenyuan; Lux, Renate

    2011-10-01

    This study investigated the bacterial communities residing in the apical portion of human teeth with apical periodontitis in primary and secondary infections by using a culture-independent molecular biology approach. Root canal samples from the apical root segments of extracted teeth were collected from 18 teeth with necrotic pulp and 8 teeth with previous endodontic treatment. Samples were processed for amplification via polymerase chain reaction and separated with denaturing gradient gel electrophoresis. Selected bands were excised from the gel and sequenced for identification. Comparable to previous studies of entire root canals, the apical bacterial communities in primary infections were significantly more diverse than in secondary infections (P = .0003). Interpatient and intrapatient comparisons exhibited similar variations in profiles. Different roots of the same teeth with secondary infections displayed low similarity in bacterial composition, whereas an equivalent sample collected from primary infection contained almost identical populations. Sequencing revealed a high prevalence of Fusobacteria, Actinomyces species, and oral Anaeroglobus geminatus in both types of infection. Many secondary infections contained Burkholderiales or Pseudomonas species, both of which represent opportunistic environmental pathogens. Certain microorganisms exhibit similar prevalence in primary and secondary infection, indicating that they are likely not eradicated during endodontic treatment. The presence of Burkholderiales and Pseudomonas species underscores the problem of environmental contamination. Treatment appears to affect the various root canals of multirooted teeth differently, resulting in local changes of the microbiota. Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  14. Evaluation of Enterococcus faecalis adhesion, penetration, and method to prevent the penetration of Enterococcus faecalis into root cementum: Confocal laser scanning microscope and scanning electron microscope analysis.

    PubMed

    Halkai, Rahul S; Hegde, Mithra N; Halkai, Kiran R

    2016-01-01

    To ascertain the role of Enterococcus faecalis in persistent infection and a possible method to prevent the penetration of E. faecalis into root cementum. One hundred and twenty human single-rooted extracted teeth divided into five groups. Group I (control): intact teeth, Group II: no apical treatment done, Group III divided into two subgroups. In Groups IIIa and IIIb, root apex treated with lactic acid of acidic and neutral pH, respectively. Group IV: apical root cementum exposed to lactic acid and roughened to mimic the apical resorption. Group V: apical treatment done same as Group IV and root-end filling done using mineral trioxide aggregate (MTA). Apical one-third of all samples immersed in E. faecalis broth for 8 weeks followed by bone morphogenetic protein and obturation and again immersed into broth for 8 weeks. Teeth split into two halves and observed under confocal laser scanning microscope and scanning electron microscope, organism identified by culture and polymerase chain reaction techniques. Adhesion and penetration was observed in Group IIIa and Group IV. Only adhesion in Group II and IIIB and no adhesion and penetration in Group I and V. Adhesion and penetration of E. faecalis into root cementum providing a long-term nidus for subsequent infection are the possible reason for persistent infection and root-end filling with MTA prevents the adhesion and penetration.

  15. Morphology of the Physiological Apical Foramen in Maxillary and Mandibular First Molars

    PubMed Central

    Abarca, J.; Zaror, C.; Monardes, H.; Hermosilla, V.; Muñoz, C.; Cantin, M.

    2015-01-01

    SUMMARY Information regarding the anatomy of the physiological apical foramen is limited. Knowing its diameter and shapes contributes to clinical work, specifically to the cleaning and shaping of the apical third. The aim of this ex vivo study was to determine the minimum and maximum diameters and shape of the physiological apical foramen in the roots of maxillary and mandibular first molars. A descriptive study was conducted on 89 recently extracted first molars. Roots 3–5 mm from the apex were sectioned and prepared for analysis at 40× magnification. The minimum and maximum diameters of each physiological foramen were measured using the program Motic Images plus 2.0 ML. The shape of the foramina, classified as round, oval or irregular, was determined by the difference between the maximum and minimum diameters. A total of 174 physiological foramina were analyzed. The average of the minimum and maximum diameters was between 0.24–0.33 mm in maxillary first molars and between 0.25–0.33 mm in mandibular first molars. In maxillary molars, the most common shape of the foramen was oval (50%), then irregular (32%), then round (18%). In mandibular molars, the oval shape was also the most frequent (59%), followed by irregular (23%) and round (18%). The findings of this study regarding the morphology of physiological apical foramina in first molars make it easier for the operator to choose the appropriately-sized instruments to perform endodontic therapy successfully. PMID:25937698

  16. Morphology of the Physiological Apical Foramen in Maxillary and Mandibular First Molars.

    PubMed

    Abarca, J; Zaror, C; Monardes, H; Hermosilla, V; Muñoz, C; Cantin, M

    2014-06-01

    Information regarding the anatomy of the physiological apical foramen is limited. Knowing its diameter and shapes contributes to clinical work, specifically to the cleaning and shaping of the apical third. The aim of this ex vivo study was to determine the minimum and maximum diameters and shape of the physiological apical foramen in the roots of maxillary and mandibular first molars. A descriptive study was conducted on 89 recently extracted first molars. Roots 3-5 mm from the apex were sectioned and prepared for analysis at 40× magnification. The minimum and maximum diameters of each physiological foramen were measured using the program Motic Images plus 2.0 ML. The shape of the foramina, classified as round, oval or irregular, was determined by the difference between the maximum and minimum diameters. A total of 174 physiological foramina were analyzed. The average of the minimum and maximum diameters was between 0.24-0.33 mm in maxillary first molars and between 0.25-0.33 mm in mandibular first molars. In maxillary molars, the most common shape of the foramen was oval (50%), then irregular (32%), then round (18%). In mandibular molars, the oval shape was also the most frequent (59%), followed by irregular (23%) and round (18%). The findings of this study regarding the morphology of physiological apical foramina in first molars make it easier for the operator to choose the appropriately-sized instruments to perform endodontic therapy successfully.

  17. Quantitative evaluation of apical extrusion of intracanal bacteria using rotary ProTaper, K3XF, twisted and hand K-file system: An ex vivo study.

    PubMed

    Ghogre, Priyanka; Chourasia, Hemant Ramesh; Agarwal, Manish; Singh, M P; Gurav, Sandeep; Ghogre, Rahul

    2015-01-01

    The aim of this study was to evaluate the number of intracanal bacteria extruded apically during root canal preparation using rotary ProTaper, K3XF, twisted, and hand K-file system. Seventy extracted single-rooted human mandibular premolar teeth were used. Access cavities were prepared and the teeth were mounted in glass vials. Root canals were then contaminated with a pure culture of Enterococcus faecalis (ATCC 29212) and incubated at 37°C for 24 h. The contaminated roots were divided into four experimental groups of 15 teeth each and one control group of 10 teeth. Group 1: ProTaper; Group 2: K3XF; Group 3: Twisted file; Group 4: Hand K-file; Group 5: Control group. Bacteria extruded from the apical foramen during instrumentation were collected into vials. The microbiological samples were incubated in culture media for 24 h. Colonies of bacteria were counted and the results were given as number of colony-forming units (CFU)/ml. The obtained data were analyzed using the Kruskal-Wallis one-way analysis of variance and Mann-Whitney U-tests. There was a significant difference between the rotary and hand instrumentation system related to the apically extruded intracanal bacteria. Both the rotary and hand instrumentation systems extruded intracanal bacteria through the apical foramen. K3XF file system showed least bacterial extrusion amongst all instrumentation groups.

  18. Par3 integrates Tiam1 and phosphatidylinositol 3-kinase signaling to change apical membrane identity

    PubMed Central

    Ruch, Travis R.; Bryant, David M.; Mostov, Keith E.; Engel, Joanne N.

    2017-01-01

    Pathogens can alter epithelial polarity by recruiting polarity proteins to the apical membrane, but how a change in protein localization is linked to polarity disruption is not clear. In this study, we used chemically induced dimerization to rapidly relocalize proteins from the cytosol to the apical surface. We demonstrate that forced apical localization of Par3, which is normally restricted to tight junctions, is sufficient to alter apical membrane identity through its interactions with phosphatidylinositol 3-kinase (PI3K) and the Rac1 guanine nucleotide exchange factor Tiam1. We further show that PI3K activity is required upstream of Rac1, and that simultaneously targeting PI3K and Tiam1 to the apical membrane has a synergistic effect on membrane remodeling. Thus, Par3 coordinates the action of PI3K and Tiam1 to define membrane identity, revealing a signaling mechanism that can be exploited by human mucosal pathogens. PMID:27881661

  19. In vitro Comparison of Apical Debris Extrusion Using Rotary and Reciprocating Systems in Severely Curved Root Canals

    PubMed Central

    Delvarani, Abbas; Mohammadzadeh Akhlaghi, Nahid; Aminirad, Raana; Tour Savadkouhi, Sohrab; Vahdati, Seyed Aliakbar

    2017-01-01

    Introduction: The aim of this in vitro study was to compare the amount of apically extruded debris after root canal preparation using rotary and reciprocating systems in severely curved root canals. Methods and Materials: Thirty six extracted human mandibular first molars with 25-35° curvature in their mesiobuccal (MB) canal (according to Schneider’s method) were cleaned and shaped with ProTaper and WaveOne systems. The extruded debris was collected and their net weight was calculated. To compare the efficiency of the two systems, the operation time was also measured. The data were analyzed with t-test. Results: The amount of extruded debris in WaveOne group was significantly greater in comparison with ProTaper group (26%). The operating time for ProTaper was however, significantly longer than WaveOne. Conclusion: Both root preparation systems caused some degree of debris extrusion through the apical foramen. However, this amount was greater in WaveOne instruments. PMID:28179921

  20. Isolation of gibberellin A8-glucoside from shoot apices of Althaea rosea.

    PubMed

    Harada, H; Yokota, T

    1970-03-01

    Gibberellin A8-glucoside has been isolated from shoot apices of Althaea rosea. It showed a weak growth-promoting activity on rice seedlings and oat mesocotyl sections but did not induce germination of lettuce seeds in darkness.

  1. Preparation Prerequisites for Effective Irrigation of Apical Root Canal: A Critical Review.

    PubMed

    Tziafas, Dimitrios; Alraeesi, Dana; Al Hormoodi, Reem; Ataya, Maamoun; Fezai, Hessa; Aga, Nausheen

    2017-10-01

    It is well recognized that disinfection of the complex root canal system at the apical root canal remains the most critical therapeutic measure to treat apical periodontitis. Observational and experimental data in relation to the anatomy of the apical root canal in different tooth types and the cross sectional diameters of the apical part of the most commonly used hand and rotary files are critically reviewed. The present data analysis confirm that the challenging issue of antibacterial efficacy of modern preparation protocols in non-surgical endodontics requires more attention to apical root canal irrigation as a balance between safety and effectiveness. Ex vivo investigations clearly indicate that a specific design of the chemo-mechanical preparation is needed at the onset of RCT, more particularly in infected teeth. Design should be based on specific anatomical parameters, and must determine the appropriate size and taper of preparation as pre-requirements for effective and safe apical irrigation. The optimal irrigation protocols might be designed on the basis of technical specifications of the preparations procedures, such as the penetration depth, the type of the needle, the required time for continuous irrigant flow, the concentration of NaOCl, and the activation parameters. Key words: Endodontics, root canal treatment, instrumentation, irrigation, apical root canal.

  2. Absorption and biological activity of phytochemical-rich extracts from açai (Euterpe oleracea Mart.) pulp and oil in vitro.

    PubMed

    Pacheco-Palencia, Lisbeth A; Talcott, Stephen T; Safe, Stephen; Mertens-Talcott, Susanne

    2008-05-28

    Polyphenolic extracts from various fruits and vegetables have been shown to exert growth inhibitory effects in cell culture studies. Whereas individual polyphenolic compounds have been extensively evaluated, understanding of the biological activity of polyphenolic extracts from natural sources is limited and critical to the understanding of their potential effects on the human body. This study investigated the absorption and antiproliferative effects of phytochemical extracts from acai pulp and a polyphenolic-enriched acai oil obtained from the fruit pulp of the acai berry ( Euterpe oleracea Mart.). Chemical composition, antioxidant properties, and polyphenolic absorption of phytochemical fractions in a Caco-2 monolayer were determined, along with their cytotoxicity in HT-29 human colon adenocarcinoma cells. Standardized extracts were characterized by their predominance of hydroxybenzoic acids, monomeric flavan-3-ols, and procyanidin dimers and trimers. Polyphenolic mixtures (0-12 microg of gallic acid equiv/mL) from both acai pulp and acai oil extracts inhibited cell proliferation by up to 90.7%, which was accompanied by an increase of up to 2.1-fold in reactive oxygen species. Absorption experiments using a Caco-2 intestinal cell monolayer demonstrated that phenolic acids such as p-hydroxybenzoic, vanillic, syringic, and ferulic acids, in the presence of DMSO, were readily transported from the apical to the basolateral side along with monomeric flavanols such as (+)-catechin and (-)-epicatechin. Results from this study provide further evidence for the bioactive properties of acai polyphenolics and offer new insight on their composition and cellular absorption.

  3. Membrane recycling at the infranuclear pole of the outer hair cell

    NASA Astrophysics Data System (ADS)

    Harasztosi, Csaba; Harasztosi, Emese; Gummer, Anthony W.

    2015-12-01

    Rapid endocytic activity of outer hair cells (OHCs) in the guinea-pig cochlea has been already studied using the fluorescent membrane marker FM1-43. It was demonstrated that vesicles were endocytosed at the apical pole of OHCs and transcytosed to the basolateral membrane and through a central strand towards the nucleus. The significance of endocytic activity in the infranuclear region is still not clear. Therefore, in this study endocytic activity at the synaptic pole of OHCs was investigated. Confocal laser scanning microscopy was used to visualize dye uptake of OHCs isolated from the guinea-pig cochlea. Signal intensity changes were quantified in the apical and basal poles relative to the signal at the membrane. Data showed no significant difference in fluorescent signal intensity changes between the opposite poles of the OHC. These results suggest that endocytic activities in both the basal and the apical poles contribute equally to the membrane recycling of OHCs.

  4. Rho-associated protein kinase regulates subcellular localisation of Angiomotin and Hippo-signalling during preimplantation mouse embryo development.

    PubMed

    Mihajlović, Aleksandar I; Bruce, Alexander W

    2016-09-01

    The differential activity of the Hippo-signalling pathway between the outer- and inner-cell populations of the developing preimplantation mouse embryo directs appropriate formation of trophectoderm and inner cell mass (ICM) lineages. Such distinct signalling activity is under control of intracellular polarization, whereby Hippo-signalling is either supressed in polarized outer cells or activated in apolar inner cells. The central role of apical-basolateral polarization to such differential Hippo-signalling regulation prompted us to reinvestigate the role of potential upstream molecular regulators affecting apical-basolateral polarity. This study reports that the chemical inhibition of Rho-associated kinase (Rock) is associated with failure to form morphologically distinct blastocysts, indicative of compromised trophectoderm differentiation, and defects in the localization of both apical and basolateral polarity factors associated with malformation of tight junctions. Moreover, Rock-inhibition mediates mislocalization of the Hippo-signalling activator Angiomotin (Amot), to the basolateral regions of outer cells and is concomitant with aberrant activation of the pathway. The Rock-inhibition phenotype is mediated by Amot, as RNAi-based Amot knockdown totally rescues the normal suppression of Hippo-signalling in outer cells. In conclusion, Rock, via regulating appropriate apical-basolateral polarization in outer cells, regulates the appropriate activity of the Hippo-signalling pathway, by ensuring correct subcellular localization of Amot protein in outer cells. Copyright © 2016 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  5. Rhubarb tannins extract inhibits the expression of aquaporins 2 and 3 in magnesium sulphate-induced diarrhoea model.

    PubMed

    Liu, Chunfang; Zheng, Yanfang; Xu, Wen; Wang, Hui; Lin, Na

    2014-01-01

    Tannins, a group of major active components of Chinese rhubarb and widely distributed in nature, have a significant antidiarrhoeal activity. Aquaporins (AQPs) 2 and 3 play important roles in regulating water transfer during diarrhoea. The present study aims to determine the effect of the total tannins extract of rhubarb on aquaporins (AQPs) 2 and 3 in diarrhoea mice and HT-29 cells both induced by magnesium sulphate (MgSO4). Our results showed that rhubarb tannins extract (RTE) significantly decreased the faecal water content in colon and evaluation index of defecation of diarrhoea mice. Interestingly, RTE could markedly reduce the mRNA and protein expression levels of AQPs 2 and 3 in apical and lateral mucosal epithelial cells in the colons of diarrhoea mice and HT-29 cells both induced by MgSO4 in a dose-dependent manner. Furthermore, RTE suppressed the production of cyclic monophosphate- (cAMP-) dependent protein kinase A catalytic subunits α (PKA C-α) and phosphorylated cAMP response element-binding protein (p-CREB, Ser133) in MgSO4-induced HT-29 cells. Our data showed for the first time that RTE inhibit AQPs 2 and 3 expression in vivo and in vitro via downregulating PKA/p-CREB signal pathway, which accounts for the antidiarrhoeal effect of RTE.

  6. Proliferation of epithelial cell rests, formation of apical cysts, and regression of apical cysts after periapical wound healing.

    PubMed

    Lin, Louis M; Huang, George T-J; Rosenberg, Paul A

    2007-08-01

    There is continuing controversy regarding the potential for inflammatory apical cysts to heal after nonsurgical endodontic therapy. Molecular cell biology may provide answers to a series of related questions. How are the epithelial cell rests of Malassez stimulated to proliferate? How are the apical cysts formed? How does the lining epithelium of apical cysts regress after endodontic therapy? Epithelial cell rests are induced to divide and proliferate by inflammatory mediators, proinflammatory cytokines, and growth factors released from host cells during periradicular inflammation. Quiescent epithelial cell rests can behave like restricted-potential stem cells if stimulated to proliferate. Formation of apical cysts is most likely caused by the merging of proliferating epithelial strands from all directions to form a three-dimensional ball mass. After endodontic therapy, epithelial cells in epithelial strands of periapical granulomas and the lining epithelium of apical cysts may stop proliferating because of a reduction in inflammatory mediators, proinflammatory cytokines, and growth factors. Epithelial cells will also regress because of activation of apoptosis or programmed cell death through deprivation of survival factors or by receiving death signals during periapical wound healing.

  7. Dental caries and pulpal disease.

    PubMed

    Zero, Domenick T; Zandona, Andrea Ferreira; Vail, Mychel Macapagal; Spolnik, Kenneth J

    2011-01-01

    This article reviews the diagnostic process, from the first clinically evident stages of the caries process to development of pulpal pathosis. The caries diagnostic process includes 4 interconnected components-staging caries lesion severity, assessing caries lesion activity, and risk assessments at the patient and tooth surface level - which modify treatment decisions for the patient. Pulpal pathosis is diagnosed as reversible pulpitis, irreversible pulpitis (asymptomatic), irreversible pulpitis (symptomatic), and pulp necrosis. Periapical disease is diagnosed as symptomatic apical periodontitis, asymptomatic apical periodontitis, acute apical abscess, and chronic apical abscess. Ultimately, the goal of any diagnosis should be to achieve better treatment decisions and health outcomes for the patient. Copyright © 2011 Elsevier Inc. All rights reserved.

  8. Nicotine-induced activation of soluble adenylyl cyclase participates in ion transport regulation in mouse tracheal epithelium.

    PubMed

    Hollenhorst, Monika I; Lips, Katrin S; Kummer, Wolfgang; Fronius, Martin

    2012-11-27

    Functional nicotinic acetylcholine receptors (nAChR) have been identified in airway epithelia and their location in the apical and basolateral membrane makes them targets for acetylcholine released from neuronal and non-neuronal sources. One function of nAChR in airway epithelia is their involvement in the regulation of transepithelial ion transport by activation of chloride and potassium channels. However, the mechanisms underlying this nicotine-induced activation of ion transport are not fully elucidated. Thus, the aim of this study was to investigate the involvement of adenylyl cyclases in the nicotine-induced ion current in mouse tracheal epithelium. To evaluate the nicotine-mediated changes of transepithelial ion transport processes electrophysiological Ussing chamber measurements were applied and nicotine-induced ion currents were recorded in the absence and presence of adenylyl cyclase inhibitors. The ion current changes induced by nicotine (100 μM, apical) were not altered in the presence of high doses of atropine (25 μM, apical and basolateral), underlining the involvement of nAChR. Experiments with the transmembrane adenylyl cyclase inhibitor 2'5'-dideoxyadenosine (50 μM, apical and basolateral) and the soluble adenylyl cyclase inhibitor KH7 (10 μM, apical and basolateral) both reduced the nicotine-mediated ion current to a similar extent. Yet, a statistically significant reduction was obtained only in the experiments with KH7. This study indicates that nicotine binding to nAChR in mouse tracheal epithelium activates transepithelial ion transport involving adenylyl cyclase activity. This might be important for novel therapeutic strategies targeting epithelial ion transport mediated by the non-neuronal cholinergic system. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Altered Regulation of Airway Epithelial Cell Chloride Channels in Cystic Fibrosis

    NASA Astrophysics Data System (ADS)

    Frizzell, Raymond A.; Rechkemmer, Gerhard; Shoemaker, Richard L.

    1986-08-01

    In many epithelial cells the chloride conductance of the apical membrane increases during the stimulation of electrolyte secretion. Single-channel recordings from human airway epithelial cells showed that β -adrenergic stimulation evoked apical membrane chloride channel activity, but this response was absent in cells from patients with cystic fibrosis (CF). However, when membrane patches were excised from CF cells into media containing sufficient free calcium (approximately 180 nanomolar), chloride channels were activated. The chloride channels of CF cells were similar to those of normal cells as judged by their current-voltage relations, ion selectivity, and kinetic behavior. These findings demonstrate the presence of chloride channels in the apical membranes of CF airway cells. Their regulation by calcium appears to be intact, but cyclic adenosine monophosphate (cAMP)-dependent control of their activity is defective.

  10. Histologic Outcomes of Uninfected Human Immature Teeth Treated with Regenerative Endodontics: 2 Case Reports.

    PubMed

    Nosrat, Ali; Kolahdouzan, Alireza; Hosseini, Farzaneh; Mehrizi, Ehsan A; Verma, Prashant; Torabinejad, Mahmoud

    2015-10-01

    A growing body of evidence exists showing the possibility of growing vital tissues in the root canal spaces of teeth with necrotic pulps and open apices. However, there is very limited histologic information regarding characteristics of tissues formed in the root canal space of human teeth after regenerative endodontics. The aim of this study was to examine clinically and histologically the outcomes of human immature teeth treated with regenerative endodontics. Two healthy birooted human maxillary first premolar teeth scheduled for extraction were included. Preoperative radiographs confirmed that these teeth had immature apices. Vitality tests showed the presence of vital pulps in these teeth. After receiving consent forms, the teeth were isolated with a rubber dam, and the pulps were completely removed. After the formation of blood clots in the canals, the teeth were covered with mineral trioxide aggregate. Four months later, the teeth were clinically and radiographically evaluated, extracted, and examined histologically. Both patients remained asymptomatic after treatment. Radiographic examination of the teeth showed signs of root development after treatment. Histologic examination of tissues growing into the root canal space of these teeth shows the presence of connective tissue, bone and cementum formation, and thickening of roots. Based on our findings, it appears that when canals of teeth with open apices are treated with regenerative endodontics, tissues of the periodontium grow into the root canals of these teeth. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  11. Mechanotransductive cascade of Myo-II-dependent mesoderm and endoderm invaginations in embryo gastrulation

    PubMed Central

    Mitrossilis, Démosthène; Röper, Jens-Christian; Le Roy, Damien; Driquez, Benjamin; Michel, Aude; Ménager, Christine; Shaw, Gorky; Le Denmat, Simon; Ranno, Laurent; Dumas-Bouchiat, Frédéric; Dempsey, Nora M.; Farge, Emmanuel

    2017-01-01

    Animal development consists of a cascade of tissue differentiation and shape change. Associated mechanical signals regulate tissue differentiation. Here we demonstrate that endogenous mechanical cues also trigger biochemical pathways, generating the active morphogenetic movements shaping animal development through a mechanotransductive cascade of Myo-II medio-apical stabilization. To mimic physiological tissue deformation with a cell scale resolution, liposomes containing magnetic nanoparticles are injected into embryonic epithelia and submitted to time-variable forces generated by a linear array of micrometric soft magnets. Periodic magnetically induced deformations quantitatively phenocopy the soft mechanical endogenous snail-dependent apex pulsations, rescue the medio-apical accumulation of Rok, Myo-II and subsequent mesoderm invagination lacking in sna mutants, in a Fog-dependent mechanotransductive process. Mesoderm invagination then activates Myo-II apical accumulation, in a similar Fog-dependent mechanotransductive process, which in turn initiates endoderm invagination. This reveals the existence of a highly dynamic self-inductive cascade of mesoderm and endoderm invaginations, regulated by mechano-induced medio-apical stabilization of Myo-II. PMID:28112149

  12. Mechanotransductive cascade of Myo-II-dependent mesoderm and endoderm invaginations in embryo gastrulation

    NASA Astrophysics Data System (ADS)

    Mitrossilis, Démosthène; Röper, Jens-Christian; Le Roy, Damien; Driquez, Benjamin; Michel, Aude; Ménager, Christine; Shaw, Gorky; Le Denmat, Simon; Ranno, Laurent; Dumas-Bouchiat, Frédéric; Dempsey, Nora M.; Farge, Emmanuel

    2017-01-01

    Animal development consists of a cascade of tissue differentiation and shape change. Associated mechanical signals regulate tissue differentiation. Here we demonstrate that endogenous mechanical cues also trigger biochemical pathways, generating the active morphogenetic movements shaping animal development through a mechanotransductive cascade of Myo-II medio-apical stabilization. To mimic physiological tissue deformation with a cell scale resolution, liposomes containing magnetic nanoparticles are injected into embryonic epithelia and submitted to time-variable forces generated by a linear array of micrometric soft magnets. Periodic magnetically induced deformations quantitatively phenocopy the soft mechanical endogenous snail-dependent apex pulsations, rescue the medio-apical accumulation of Rok, Myo-II and subsequent mesoderm invagination lacking in sna mutants, in a Fog-dependent mechanotransductive process. Mesoderm invagination then activates Myo-II apical accumulation, in a similar Fog-dependent mechanotransductive process, which in turn initiates endoderm invagination. This reveals the existence of a highly dynamic self-inductive cascade of mesoderm and endoderm invaginations, regulated by mechano-induced medio-apical stabilization of Myo-II.

  13. Position-dependent patterning of spontaneous action potentials in immature cochlear inner hair cells

    PubMed Central

    Johnson, Stuart L.; Eckrich, Tobias; Kuhn, Stephanie; Zampini, Valeria; Franz, Christoph; Ranatunga, Kishani M.; Roberts, Terri P.; Masetto, Sergio; Knipper, Marlies; Kros, Corné J.; Marcotti, Walter

    2011-01-01

    Spontaneous action potential activity is crucial for mammalian sensory system development. In the auditory system, patterned firing activity has been observed in immature spiral ganglion cells and brain-stem neurons and is likely to depend on cochlear inner hair cell (IHC) action potentials. It remains uncertain whether spiking activity is intrinsic to developing IHCs and whether it shows patterning. We found that action potentials are intrinsically generated by immature IHCs of altricial rodents and that apical IHCs exhibit bursting activity as opposed to more sustained firing in basal cells. We show that the efferent neurotransmitter ACh, by fine-tuning the IHC’s resting membrane potential (Vm), is crucial for the bursting pattern in apical cells. Endogenous extracellular ATP also contributes to the Vm of apical and basal IHCs by activating SK2 channels. We hypothesize that the difference in firing pattern along the cochlea instructs the tonotopic differentiation of IHCs and auditory pathway. PMID:21572434

  14. Position-dependent patterning of spontaneous action potentials in immature cochlear inner hair cells.

    PubMed

    Johnson, Stuart L; Eckrich, Tobias; Kuhn, Stephanie; Zampini, Valeria; Franz, Christoph; Ranatunga, Kishani M; Roberts, Terri P; Masetto, Sergio; Knipper, Marlies; Kros, Corné J; Marcotti, Walter

    2011-06-01

    Spontaneous action potential activity is crucial for mammalian sensory system development. In the auditory system, patterned firing activity has been observed in immature spiral ganglion and brain-stem neurons and is likely to depend on cochlear inner hair cell (IHC) action potentials. It remains uncertain whether spiking activity is intrinsic to developing IHCs and whether it shows patterning. We found that action potentials were intrinsically generated by immature IHCs of altricial rodents and that apical IHCs showed bursting activity as opposed to more sustained firing in basal cells. We show that the efferent neurotransmitter acetylcholine fine-tunes the IHC's resting membrane potential (V(m)), and as such is crucial for the bursting pattern in apical cells. Endogenous extracellular ATP also contributes to the V(m) of apical and basal IHCs by triggering small-conductance Ca(2+)-activated K(+) (SK2) channels. We propose that the difference in firing pattern along the cochlea instructs the tonotopic differentiation of IHCs and auditory pathway.

  15. Release of Apical Dominance in Potato Tuber Is Accompanied by Programmed Cell Death in the Apical Bud Meristem[C][W

    PubMed Central

    Teper-Bamnolker, Paula; Buskila, Yossi; Lopesco, Yael; Ben-Dor, Shifra; Saad, Inbal; Holdengreber, Vered; Belausov, Eduard; Zemach, Hanita; Ori, Naomi; Lers, Amnon; Eshel, Dani

    2012-01-01

    Potato (Solanum tuberosum) tuber, a swollen underground stem, is used as a model system for the study of dormancy release and sprouting. Natural dormancy release, at room temperature, is initiated by tuber apical bud meristem (TAB-meristem) sprouting characterized by apical dominance (AD). Dormancy is shortened by treatments such as bromoethane (BE), which mimics the phenotype of dormancy release in cold storage by inducing early sprouting of several buds simultaneously. We studied the mechanisms governing TAB-meristem dominance release. TAB-meristem decapitation resulted in the development of increasing numbers of axillary buds with time in storage, suggesting the need for autonomous dormancy release of each bud prior to control by the apical bud. Hallmarks of programmed cell death (PCD) were identified in the TAB-meristems during normal growth, and these were more extensive when AD was lost following either extended cold storage or BE treatment. Hallmarks included DNA fragmentation, induced gene expression of vacuolar processing enzyme1 (VPE1), and elevated VPE activity. VPE1 protein was semipurified from BE-treated apical buds, and its endogenous activity was fully inhibited by a cysteinyl aspartate-specific protease-1-specific inhibitor N-Acetyl-Tyr-Val-Ala-Asp-CHO (Ac-YVAD-CHO). Transmission electron microscopy further revealed PCD-related structural alterations in the TAB-meristem of BE-treated tubers: a knob-like body in the vacuole, development of cytoplasmic vesicles, and budding-like nuclear segmentations. Treatment of tubers with BE and then VPE inhibitor induced faster growth and recovered AD in detached and nondetached apical buds, respectively. We hypothesize that PCD occurrence is associated with the weakening of tuber AD, allowing early sprouting of mature lateral buds. PMID:22362870

  16. Adenovirus Entry From the Apical Surface of Polarized Epithelia Is Facilitated by the Host Innate Immune Response

    PubMed Central

    Kotha, Poornima L. N.; Sharma, Priyanka; Kolawole, Abimbola O.; Yan, Ran; Alghamri, Mahmoud S.; Brockman, Trisha L.; Gomez-Cambronero, Julian; Excoffon, Katherine J. D. A.

    2015-01-01

    Prevention of viral-induced respiratory disease begins with an understanding of the factors that increase or decrease susceptibility to viral infection. The primary receptor for most adenoviruses is the coxsackievirus and adenovirus receptor (CAR), a cell-cell adhesion protein normally localized at the basolateral surface of polarized epithelia and involved in neutrophil transepithelial migration. Recently, an alternate isoform of CAR, CAREx8, has been identified at the apical surface of polarized airway epithelia and is implicated in viral infection from the apical surface. We hypothesized that the endogenous role of CAREx8 may be to facilitate host innate immunity. We show that IL-8, a proinflammatory cytokine and a neutrophil chemoattractant, stimulates the protein expression and apical localization of CAREx8 via activation of AKT/S6K and inhibition of GSK3β. Apical CAREx8 tethers infiltrating neutrophils at the apical surface of a polarized epithelium. Moreover, neutrophils present on the apical-epithelial surface enhance adenovirus entry into the epithelium. These findings suggest that adenovirus evolved to co-opt an innate immune response pathway that stimulates the expression of its primary receptor, apical CAREx8, to allow the initial infection the intact epithelium. In addition, CAREx8 is a new target for the development of novel therapeutics for both respiratory inflammatory disease and adenoviral infection. PMID:25768646

  17. Chloride channel inhibition by a red wine extract and a synthetic small molecule prevents rotaviral secretory diarrhoea in neonatal mice.

    PubMed

    Ko, Eun-A; Jin, Byung-Ju; Namkung, Wan; Ma, Tonghui; Thiagarajah, Jay R; Verkman, A S

    2014-07-01

    Rotavirus is the most common cause of severe secretory diarrhoea in infants and young children globally. The rotaviral enterotoxin, NSP4, has been proposed to stimulate calcium-activated chloride channels (CaCC) on the apical plasma membrane of intestinal epithelial cells. We previously identified red wine and small molecule CaCC inhibitors. To investigate the efficacy of a red wine extract and a synthetic small molecule, CaCCinh-A01, in inhibiting intestinal CaCCs and rotaviral diarrhoea. Inhibition of CaCC-dependent current was measured in T84 cells and mouse ileum. The effectiveness of an orally administered wine extract and CaCCinh-A01 in inhibiting diarrhoea in vivo was determined in a neonatal mouse model of rotaviral infection. Screening of ∼150 red wines revealed a Cabernet Sauvignon that inhibited CaCC current in T84 cells with IC50 at a ∼1:200 dilution, and higher concentrations producing 100% inhibition. A >1 kdalton wine extract prepared by dialysis, which retained full inhibition activity, blocked CaCC current in T84 cells and mouse intestine. In rotavirus-inoculated mice, oral administration of the wine extract prevented diarrhoea by inhibition of intestinal fluid secretion without affecting rotaviral infection. The wine extract did not inhibit the cystic fibrosis chloride channel (CFTR) in cell cultures, nor did it prevent watery stools in neonatal mice administered cholera toxin, which activates CFTR-dependent fluid secretion. CaCCinh-A01 also inhibited rotaviral diarrhoea. Our results support a pathogenic role for enterocyte CaCCs in rotaviral diarrhoea and demonstrate the antidiarrhoeal action of CaCC inhibition by an alcohol-free, red wine extract and by a synthetic small molecule. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  18. Role of PIN-mediated auxin efflux in apical hook development of Arabidopsis thaliana.

    PubMed

    Zádníková, Petra; Petrásek, Jan; Marhavy, Peter; Raz, Vered; Vandenbussche, Filip; Ding, Zhaojun; Schwarzerová, Katerina; Morita, Miyo T; Tasaka, Masao; Hejátko, Jan; Van Der Straeten, Dominique; Friml, Jirí; Benková, Eva

    2010-02-01

    The apical hook of dark-grown Arabidopsis seedlings is a simple structure that develops soon after germination to protect the meristem tissues during emergence through the soil and that opens upon exposure to light. Differential growth at the apical hook proceeds in three sequential steps that are regulated by multiple hormones, principally auxin and ethylene. We show that the progress of the apical hook through these developmental phases depends on the dynamic, asymmetric distribution of auxin, which is regulated by auxin efflux carriers of the PIN family. Several PIN proteins exhibited specific, partially overlapping spatial and temporal expression patterns, and their subcellular localization suggested auxin fluxes during hook development. Genetic manipulation of individual PIN activities interfered with different stages of hook development, implying that specific combinations of PIN genes are required for progress of the apical hook through the developmental phases. Furthermore, ethylene might modulate apical hook development by prolonging the formation phase and strongly suppressing the maintenance phase. This ethylene effect is in part mediated by regulation of PIN-dependent auxin efflux and auxin signaling.

  19. Comparison of apical debris extrusion of two rotary systems and one reciprocating system

    PubMed Central

    Vivekanandhan, Paramasivam; Subbiya, Arunajatesan; Mitthra, Suresh; Karthick, Arumugam

    2016-01-01

    Background: Postoperative inflammation and endodontic failure occur due to apical extrusion of debris. Aim: To evaluate the amount of apically extruded debris after preparation of straight root canals in extracted human teeth using three rotary systems. Materials and Methods: Sixty single-rooted human teeth with single root canal and apical foramen were selected and randomly divided into three experimental groups (n = 20) according to the rotary system used: Group 1 - ProTaper, Group 2 - Revo-S, Group 3 - WaveOne. Apical enlargement was done up to size 40/0.06 for Groups 1 and 2 and 40/0.08 for Group 3. For debris collection, each tooth was held in a preweighed Eppendorf tube fixed inside a glass vial through rubber plug. After instrumentation, each tooth was separated from the Eppendorf tube and the debris adhering to the root surface was collected by washing the root with 1 mL of bi-distilled water and incubated at 70°C for 5 days. The dry weight of extruded debris was weighed in an electronic balance. Statistical Analysis: The mean weights of extruded debris were statistically analyzed using GraphPad prism version 6 and the intergroup comparison was done using unpaired t-test. Results: There was a statistically significant difference in the mean debris score among all the three groups - ProTaper (0.00065 g), Revo-S (0.00045 g) and WaveOne (0.00089 g). Conclusion: Revo-S system was associated with significantly less debris extrusion compared with ProTaper or WaveOne. PMID:27217638

  20. Incidence of Apical Crack Initiation during Canal Preparation using Hand Stainless Steel (K-File) and Hand NiTi (Protaper) Files.

    PubMed

    Soni, Dileep; Raisingani, Deepak; Mathur, Rachit; Madan, Nidha; Visnoi, Suchita

    2016-01-01

    To evaluate the incidence of apical crack initiation during canal preparation with stainless steel K-files and hand protaper files (in vitro study). Sixty extracted mandibular premo-lar teeth are randomly selected and embedded in an acrylic tube filled with autopolymerizing resin. A baseline image of the apical surface of each specimen was recorded under a digital microscope (80×). The cervical and middle thirds of all samples were flared with #2 and #1 Gates-Glidden (GG) drills, and a second image was recorded. The teeth were randomly divided into four groups of 15 teeth each according to the file type (hand K-file and hand-protaper) and working length (WL) (instrumented at WL and 1 mm less than WL). Final image after dye penetration and photomicrograph of the apical root surface were digitally recorded. Maximum numbers of cracks were observed with hand protaper files compared with hand K-file at the WL and 1 mm short of WL. Chi-square testing revealed a highly significant effect of WL on crack formation at WL and 1 mm short of WL (p = 0.000). Minimum numbers of cracks at WL and 1 mm short of WL were observed with hand K-file and maximum with hand protaper files. Soni D, Raisingani D, Mathur R, Madan N, Visnoi S. Incidence of Apical Crack Initiation during Canal Preparation using Hand Stainless Steel (K-File) and Hand NiTi (Protaper) Files. Int J Clin Pediatr Dent 2016;9(4):303-307.

  1. Sealing ability of grar MTA AngelusTM, CPM TM and MBPc used as apical plugs.

    PubMed

    Orosco, Fernando Accorsi; Bramante, Clovis Monteiro; Garcia, Roberto Brandão; Bernadineli, Norberti; Moraes, Ivaldo Gomes de

    2008-01-01

    This study evaluated the sealing ability of apical plugs fabricated with gray MTA Angelus sealer, CPM TM sealer and MBPc sealer. The root canals of 98 extracted single-rooted human teeth were instrumented with #5 to #1 Gates Glidden drills according to the crown-down technique until the #1 drill could pass through the apical foramen. The specimens were then prepared with K-files, starting with an ISO 50 until an ISO 90 could be visualized 1 mm beyond the apex. After root canal preparation, the external surface of each root was rendered impermeable and roots were assigned to 3 experimental groups (n = 30), which received a 5-mm thick apical plug of gray MTA Angelus, CPM and MBPc, and two control groups (n=4). The remaining portion of the canal in the experimental groups was filled by the lateral condensation technique. The teeth of each group, properly identified, were fixed on utility wax by their crowns and were placed in plastic flasks, leaving the apex free and facing upward. The flasks were filled with 0.2% Rhodamine B solution, pH 7.0, so as to completely cover the root apex of all teeth. The sealing ability was analyzed by measuring 0.2% Rhodamine B leakage after all groups had been maintained in this solution for 48 hours. Data were analyzed statistically by Kruskal-Wallis test and Dunn test with a=5%. The results showed that, among the tested materials used for fabrication of apical plugs, MBPc sealer had the least amount of leakage with statistically significant difference (p<0.05).

  2. SEALING ABILITY OF GRAY MTA ANGELUS™, CPM™ AND MBPC USED AS APICAL PLUGS

    PubMed Central

    Orosco, Fernando Accorsi; Bramante, Clovis Monteiro; Garcia, Roberto Brandão; Bernardineli, Norberti; de Moraes, Ivaldo Gomes

    2008-01-01

    This study evaluated the sealing ability of apical plugs fabricated with gray MTA Angelus™ sealer, CPM™ sealer and MBPc sealer. The root canals of 98 extracted single-rooted human teeth were instrumented with #5 to #1 Gates Glidden drills according to the crown-down technique until the #1 drill could pass through the apical foramen. The specimens were then prepared with K-files, starting with an ISO 50 until an ISO 90 could be visualized 1 mm beyond the apex. After root canal preparation, the external surface of each root was rendered impermeable and roots were assigned to 3 experimental groups (n = 30), which received a 5-mm thick apical plug of gray MTA Angelus™, CPM ™ and MBPc, and two control groups (n=4). The remaining portion of the canal in the experimental groups was filled by the lateral condensation technique. The teeth of each group, properly identified, were fixed on utility wax by their crowns and were placed in plastic flasks, leaving the apex free and facing upward. The flasks were filled with 0.2% Rhodamine B solution, pH 7.0, so as to completely cover the root apex of all teeth. The sealing ability was analyzed by measuring 0.2% Rhodamine B leakage after all groups had been maintained in this solution for 48 hours. Data were analyzed statistically by Kruskal-Wallis test and Dunn test with a=5%. The results showed that, among the tested materials used for fabrication of apical plugs, MBPc sealer had the least amount of leakage with statistically significant difference (p<0.05). PMID:19089289

  3. Effects of increased apical enlargement on the amount of unprepared areas and coronal dentine removal: a micro-computed tomography study.

    PubMed

    Pérez, A R; Alves, F R F; Marceliano-Alves, M F; Provenzano, J C; Gonçalves, L S; Neves, A A; Siqueira, J F

    2018-06-01

    To evaluate the effects of progressive apical enlargement on the amount of unprepared root canal surface area and remaining dentine thickness. The root canals of 30 extracted mandibular incisors with Vertucci's type I configuration were instrumented with rotary HyFlex CM instruments (Coltene-Whaledent, Altstätten, Switzerland) up to 4 instruments larger than the first one that bound at the working length (WL). Teeth were scanned in a micro-computed tomography (micro-CT) device before canal preparation and after instrumentation with the 2nd, 3rd and 4th larger instruments. The amount of unprepared surface area in the full canal or in the apical 4 mm as well as the remaining dentine thickness at 10 mm from the WL were calculated and compared. The general linear model for repeated measures adjusted by Bonferroni's post hoc test was used for statistic analysis. There was a significant reduction in the amount of unprepared areas after each increase in preparation size (P < 0.01). This was observed for both the full canal length and the 4-mm apical segment. The amount of remaining dentine was also significantly reduced after each file size (P < 0.01). However, dentine thickness always remained greater than 1 mm, even after using the largest instrument. Apical preparations up to 4 instruments larger than the first one to bind at the WL caused a significant progressive reduction in the unprepared canal area. © 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  4. Evaluation of apically extruded debris during root canal retreatment with two different rotary systems followed by a self-adjusting file.

    PubMed

    Cakici, Fatih; Cakici, Elif B; Küçükekenci, Funda Fundaoglu

    2016-02-01

    To compare the amount of apically extruded debris during root canal retreatment using ProTaper retreatment system (Dentsply Maillefer, Ballaigues, Switzerland), ProTaper retreatment system with Self-Adjusting File (SAF) system (ReDent-Nova, Ra'anana, Israel), Mtwo retreatment system (VDW, Munich, Germany), Mtwo retreatment system with SAF instruments. In total, 72 extracted human mandibular incisor teeth were used. All root canals were prepared with ProTaper universal (Dentsply Maillefer) up to F2 file and filled with gutta percha and AH plus sealer using cold lateral condensation before being assembled randomly into 4 groups (n = 18 each). Root canal filling materials were removed using the ProTaper retreatment system, the ProTaper retreatment system followed by SAF system, Mtwo retreatment system and Mtwo retreatment system followed by SAF system. Debris extruded apically during the removal of canal filling material was collected into preweighed Eppendorf tubes. The tubes were then stored in an incubator at 70°C for 5 days. The weight of the dry extruded debris was established by subtracting the preretreatment and postretreatment weight of the Eppendorf tubes for each group. The data obtained were analyzed using Kruskal-Wallis test. All retreatment techniques caused the apical extrusion of debris. There was no significant difference between the groups statistically (p>0.05). The results of this study showed that SAF system after Mtwo retreatment system and ProTaper retreatment system for improving retreatment has no significant effect on the amount of debris extruded apically.

  5. Solubilization methods and reference 2-DE map of cow milk fat globules.

    PubMed

    Bianchi, Laura; Puglia, Michele; Landi, Claudia; Matteoni, Silvia; Perini, Daniele; Armini, Alessandro; Verani, Margherita; Trombetta, Claudia; Soldani, Patrizia; Roncada, Paola; Greppi, Gianfranco; Pallini, Vitaliano; Bini, Luca

    2009-07-21

    Milk fat globules (MFGs) are secretory vesicles assembled and secreted by mammary epithelial cells during lactation. They consist of fat globules surrounded by a lipid bilayer membrane which is derived from the apical membrane of the lactating cells. MFGs contain, besides lipids, proteins from the apical plasma membrane and from the cytoplasmatic material. Their peculiar vesicle nature makes them a suitable and easily available source of biological material in monitoring the physiopathological state of the mammary gland. Unfortunately, the conspicuous lipidic component of MFGs consistently limits protein extraction and purification for MFG proteomic investigations. This work deals with the development of a suitable procedure for protein extraction from the cow MFGs in order to qualitatively and quantitatively improve 2-D electropherograms of the MFG. MFGs were purified from raw milk by centrifugation and then delipidated/precipitated. The resulting protein pellets were solubilised using four different 2-D SDS PAGE compatible lysis buffers. Applied methodological procedures for protein extraction and evaluation of the resulting 2-D protein-pattern are presented and discussed. Using these procedures a reference 2-D map of cow milk fat globules is also reported. The majority of the obtained identifications was represented by proteins involved in lipid synthesis or in fat globule secretion.

  6. Visualization of removal of trapped air from the apical region in simulated root canals by laser-activated irrigation using an Er,Cr:YSGG laser.

    PubMed

    Peeters, Harry Huiz; De Moor, Roeland J G; Suharto, Djoko

    2015-08-01

    The aim of this visualization study was to obtain a better understanding of the mechanism by which trapped air is removed from the apical region of simulated root canals by activation of an irrigant using an erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser during endodontic procedures. A high-speed imaging system with high temporal and spatial resolution was used to visualize laser-induced shock waves in a resin block model with a curved root canal (inner diameter at the apex 0.08 mm, taper 4 %, crown height 10 mm, overall length 40 mm) and a glass cylinder model with a straight root canal (inner diameter 1 mm, crown height 10 mm, overall length 40 mm). The study utilized MZ3 and RFT3 tips in each model, without water or air spray, and with an average power of 1 W at 35 Hz. Laser-activated irrigation overcame the airlock effect by releasing air trapped in the air column. The mechanism underlying the removal of trapped air from the apical region using an Er,Cr:YSGG laser in a dry root canal is via the disruption of the surface tension at the solution-air interface. This disruption, caused by bubble implosion (cavitation), displaces air in the form of bubbles from the apical region toward the solution, which allows the solution to travel apically.

  7. Sodium hyaluronate accelerates the healing process in tooth sockets of rats.

    PubMed

    Mendes, Renato M; Silva, Gerluza A B; Lima, Miguel F; Calliari, Marcelo V; Almeida, Alvair P; Alves, José B; Ferreira, Anderson J

    2008-12-01

    In this study we evaluated the effects of sodium hyaluronate (HY) in the healing process of tooth sockets of rats. Immediately after the extraction of the upper first molars of male Holtzman rats, right sockets were treated with 1% HY gel (approximately 0.1 ml), while left sockets were used as control (blood clot). The animals were sacrificed at 2, 7, and 21 days after tooth extraction and upper maxillaries processed for histological and morphometric analysis of the apical and medium thirds of the sockets. Carbopol, an inert gel, was used to evaluate the mechanical effect of gel injection into sockets. Expression of bone morphogenetic protein-2 (BMP-2) and osteopontin (OPN) was determined by immunohistochemistry at 1, 2, 3, 4, 5, and 7 days after tooth extraction. Histological analysis showed that HY treatment induced earlier trabecular bone deposition resulting in a bone matrix more organized at 7 and 21 days after tooth extraction. Also, HY elicited significant increase in the amount of bone trabeculaes at 7 and 21 days after tooth extraction (percentage of trabecular bone area at 7 days: 13.21+/-4.66% vs. 2.58+/-1.36% in the apical third of control sockets) and in the vessels counting at 7 days. Conversely, the number of cell nuclei was decreased in HY-treated sockets. Additionally, expression of BMP-2 and OPN was enhanced in HY-treated sockets compared with control sockets. These findings suggest that HY accelerates the healing process in tooth sockets of rats stimulating the expression of osteogenic proteins.

  8. Enhanced resistance to soybean cyst nematode Heterodera glycines in transgenic soybean by silencing putative CLE receptors

    USDA-ARS?s Scientific Manuscript database

    CLE peptides are small extracellular proteins important in regulating plant meristematic activity through the CLE-receptor kinase-WOX signaling module. Stem cell pools in the SAM (shoot apical meristem), RAM (root apical meristem), and vascular cambium are tightly controlled by CLE signaling pathway...

  9. Recycling Endosomes of Polarized Epithelial Cells Actively Sort Apical and Basolateral Cargos into Separate Subdomains

    PubMed Central

    Thompson, Anthony; Nessler, Randy; Wisco, Dolora; Anderson, Eric; Winckler, Bettina

    2007-01-01

    The plasma membranes of epithelial cells plasma membranes contain distinct apical and basolateral domains that are critical for their polarized functions. However, both domains are continuously internalized, with proteins and lipids from each intermixing in supranuclear recycling endosomes (REs). To maintain polarity, REs must faithfully recycle membrane proteins back to the correct plasma membrane domains. We examined sorting within REs and found that apical and basolateral proteins were laterally segregated into subdomains of individual REs. Subdomains were absent in unpolarized cells and developed along with polarization. Subdomains were formed by an active sorting process within REs, which precedes the formation of AP-1B–dependent basolateral transport vesicles. Both the formation of subdomains and the fidelity of basolateral trafficking were dependent on PI3 kinase activity. This suggests that subdomain and transport vesicle formation occur as separate sorting steps and that both processes may contribute to sorting fidelity. PMID:17494872

  10. The RCN1-encoded A subunit of protein phosphatase 2A increases phosphatase activity in vivo

    NASA Technical Reports Server (NTRS)

    Deruere, J.; Jackson, K.; Garbers, C.; Soll, D.; Delong, A.; Evans, M. L. (Principal Investigator)

    1999-01-01

    Protein phosphatase 2A (PP2A), a heterotrimeric serine/threonine-specific protein phosphatase, comprises a catalytic C subunit and two distinct regulatory subunits, A and B. The RCN1 gene encodes one of three A regulatory subunits in Arabidopsis thaliana. A T-DNA insertion mutation at this locus impairs root curling, seedling organ elongation and apical hypocotyl hook formation. We have used in vivo and in vitro assays to gauge the impact of the rcn1 mutation on PP2A activity in seedlings. PP2A activity is decreased in extracts from rcn1 mutant seedlings, and this decrease is not due to a reduction in catalytic subunit expression. Roots of mutant seedlings exhibit increased sensitivity to the phosphatase inhibitors okadaic acid and cantharidin in organ elongation assays. Shoots of dark-grown, but not light-grown seedlings also show increased inhibitor sensitivity. Furthermore, cantharidin treatment of wild-type seedlings mimics the rcn1 defect in root curling, root waving and hypocotyl hook formation assays. In roots of wild-type seedlings, RCN1 mRNA is expressed at high levels in root tips, and accumulates to lower levels in the pericycle and lateral root primordia. In shoots, RCN1 is expressed in the apical hook and the basal, rapidly elongating cells in etiolated hypocotyls, and in the shoot meristem and leaf primordia of light-grown seedlings. Our results show that the wild-type RCN1-encoded A subunit functions as a positive regulator of the PP2A holoenzyme, increasing activity towards substrates involved in organ elongation and differential cell elongation responses such as root curling.

  11. Polyalkoxybenzenes from plants. 5. Parsley seed extract in synthesis of azapodophyllotoxins featuring strong tubulin destabilizing activity in the sea urchin embryo and cell culture assays.

    PubMed

    Semenova, Marina N; Kiselyov, Alex S; Tsyganov, Dmitry V; Konyushkin, Leonid D; Firgang, Sergei I; Semenov, Roman V; Malyshev, Oleg R; Raihstat, Mikhail M; Fuchs, Fabian; Stielow, Anne; Lantow, Margareta; Philchenkov, Alex A; Zavelevich, Michael P; Zefirov, Nikolay S; Kuznetsov, Sergei A; Semenov, Victor V

    2011-10-27

    A series of 4-azapodophyllotoxin derivatives with modified rings B and E have been synthesized using allylpolyalkoxybenzenes from parsley seed oil. The targeted molecules were evaluated in vivo in a phenotypic sea urchin embryo assay for antimitotic and tubulin destabilizing activity. The most active compounds identified by the in vivo sea urchin embryo assay featured myristicin-derived ring E. These molecules were determined to be more potent than podophyllotoxin. Cytotoxic effects of selected molecules were further confirmed and evaluated by conventional assays with A549 and Jurkat human leukemic T-cell lines including cell growth inhibition, cell cycle arrest, cellular microtubule disruption, and induction of apoptosis. The ring B modification yielded 6-OMe substituted molecule as the most active compound. Finally, in Jurkat cells, compound induced caspase-dependent apoptosis mediated by the apical caspases-2 and -9 and not caspase-8, implying the involvement of the intrinsic caspase-9-dependent apoptotic pathway.

  12. A comparative evaluation of gutta percha removal and extrusion of apical debris by rotary and hand files.

    PubMed

    Chandrasekar; Ebenezar, A V Rajesh; Kumar, Mohan; Sivakumar, A

    2014-11-01

    The aim of this study was to evaluate the efficacy of Protaper retreatment files in comparison with RaCe, K3 and H-files for removal of gutta-percha and apically extruded debris using volumetric analysis. Forty extracted single rooted maxillary incisor teeth with straight canals and mature apices were selected for the study. After access cavity preparation, apical patency was confirmed with a size 10 K-file extending 1mm beyond the point at which it was first visible at the apical end. Working lengths were determined with the use of size 15 K-file. The canals were prepared in a step-back technique and the master apical file was size 30 for all teeth. 3% sodium hypochlorite was used as an irrigant after each instrumentation. Before final rinse, size 20 K-file was passed 1mm beyond the apex to remove any dentinal shaving plugs and maintain the apical patency. Then the canals were dried with paper points. The root canal was filled using standard gutta-percha points and zinc oxide eugenol sealer under lateral condensation technique. The teeth were then randomly divided into four groups of ten teeth each based on the instrument used for gutta percha removal. All the rotary instruments used in this study were rotated at 300rpm. The instruments used were: Group 1 - RaCe Files, Group 2 - ProTaper retreatment Files, Group 3 - K3 Files and Group 4 - H Files. The volume of the obturating material was calculated before and after removal using volumetric analysis with spiral CT. The removal efficacy with each instrument was calculated and statistically analysed. The results of the study show that the ProTaper retreatment files (Group 2) (97.4%) showed the highest efficiency in the removal of obturating material, which was followed by RaCe (95.74%), K3 (92.86%) and H files (90.14%) with the efficiency in the decreasing order. Similarly the mean apical extrusion in H files (0.000 ± 0.002) was significantly lower than all the rotary instruments. However, the difference among the rotary files were not statistically significant (p>0.05). ProTaper retreatment files show significant difference over other groups in removal of obturating material and can be the system of choice in endodontic retreatment procedures.

  13. Toll-like receptor 2-mediated peptidoglycan uptake by immature intestinal epithelial cells from apical side and exosome-associated transcellular transcytosis

    PubMed Central

    Bu, Heng-Fu; Wang, Xiao; Tang, Yi; Koti, Viola; Tan, Xiao-Di

    2015-01-01

    Peptidoglycan is a potent immune adjuvant derived from bacterial cell walls. Previous investigations suggest that intestinal epithelium may absorb peptidoglycan from the lumen. Nonetheless, how peptidoglycan is taken up and crosses intestinal epithelium remains largely unclear. Here, we first characterized peptidoglycan transport in vitro using IEC-18 and HT29-CL19A cells, which represent less mature epithelial cells in intestinal crypts. With fluorescent microscopy, we visualized internalization of dual-labeled peptidoglycan by enterocytes. Engulfed peptidoglycan was found to form a complex with peptidoglycan recognition protein-3, which may facilitate delivering peptidoglycan in vivo. Utilizing electronic microscopy, we revealed that uptake of apical peptidoglycan across intestinal epithelial monolayers was involved in phagocytosis, multivesicular body formation, and exosome secretion. We also studied transport of peptidoglycan using the transwell system. Our data indicated that apically loaded peptidoglycan was exocytosed to the basolateral compartment with exosomes by HT29-CL19A cells. The peptidoglycan-contained basolateral exosome extracts induced macrophage activation. Through gavaging mice with labeled peptidoglycan, we found that luminal peptidoglycan was taken up by columnar epithelial cells in crypts of the small intestine. Furthermore, we showed that pre-confluent immature but not post-confluent mature C2BBe1 cells engulfed peptidoglycan via a toll-like receptor 2-dependent manner. Together, our findings suggest that (1) crypt-based immature intestinal epithelial cells play an important role in transport of luminal peptidoglycan over the intestinal epithelium; and (2) luminal peptidoglycan is transcytosed across intestinal epithelia via a toll-like receptor 2-meciated phagocytosis-multivesicular body-exosome pathway. The absorbed peptidoglycan and its derivatives may facilitate maintenance of intestinal immune homeostasis. PMID:20020500

  14. Comparison between high concentration EDTA (24%) and low concentration EDTA (3%) with surfactant upon removal of smear layer after rotary instrumentation: a SEM study.

    PubMed

    Daghustani, Mohsen; Alhammadi, Ahmad; Merdad, Khalid; Ohlin, Johan; Erhardt, Fredrik; Ahlquist, Michael

    2011-01-01

    This in vitro study compare cleanliness of tooth canal walls regarding smear layer after final treatment with 24% ethylenediaminetetraaceticacid (EDTA) and 3% EDTA with or without surfactant. Sixty extracted teeth, randomly distributed into four groups, were prepared using ProFile instruments (DENTSPLY, Maillefer, Ballaigues, Switzerland), and subjected to different final irrigation solutions: group A, 24% EDTA; group B, 3% EDTA with surfactant; group C (positive control), 3% EDTA; and group D (negative control), 0.5% sodium hypochlorite. Roots were sectioned, examined and evaluated under scanning electron microscope; microphotographs were taken for the coronal, middle and apical third of each specimen. Statistical analysis showed no difference regarding presence of smear layer between test groups in the coronal and apical sections. They were cleaned in the coronal sections and uncleaned in the apical sections. In the middle section, group B was significantly cleaner (p < 0.05) than the other groups. In conclusion, surfactant in combination with EDTA did not improve root canal cleanliness and there is no difference between different EDTA concentrations in removing the smear layer.

  15. Shaping abilities of two different engine-driven rotary nickel titanium systems or stainless steel balanced-force technique in mandibular molars.

    PubMed

    Matwychuk, Michael J; Bowles, Walter R; McClanahan, Scott B; Hodges, Jim S; Pesun, Igor J

    2007-07-01

    The purpose of this study was to compare apical transportation, working-length changes, and instrumentation time by using nickel-titanium (Ni-Ti) rotary file systems (crown-down method) or stainless steel hand files (balanced-force technique) in mesiobuccal canals of extracted mandibular molars. The curvature of each canal was determined and teeth placed into three equivalent groups. Group 1 was instrumented with Sequence (Brasseler USA, Savannah, GA) rotary files, group 2 with Liberator (Miltex Inc, York, PA) rotary files, and group 3 with Flex-R (Union Broach, New York, NY) files. Pre- and postoperative radiographs were superimposed to measure loss of working length and apical transportation as shown by changes in radius of curvature and the long-axis canal angle. Sequence rotary files, Liberator rotary files, and Flex-R hand files had similar effects on apical canal transportation and changes in working length, with no significant differences detected among the 3 groups. Hand instrumentation times were longer than with either Ni-Ti rotary group, whereas the rotary NiTi groups had a higher incidence of fracture.

  16. RhoA GTPase inhibition organizes contraction during epithelial morphogenesis

    PubMed Central

    Mason, Frank M.; Xie, Shicong; Vasquez, Claudia G.; Tworoger, Michael

    2016-01-01

    During morphogenesis, contraction of the actomyosin cytoskeleton within individual cells drives cell shape changes that fold tissues. Coordination of cytoskeletal contractility is mediated by regulating RhoA GTPase activity. Guanine nucleotide exchange factors (GEFs) activate and GTPase-activating proteins (GAPs) inhibit RhoA activity. Most studies of tissue folding, including apical constriction, have focused on how RhoA is activated by GEFs to promote cell contractility, with little investigation as to how GAPs may be important. Here, we identify a critical role for a RhoA GAP, Cumberland GAP (C-GAP), which coordinates with a RhoA GEF, RhoGEF2, to organize spatiotemporal contractility during Drosophila melanogaster apical constriction. C-GAP spatially restricts RhoA pathway activity to a central position in the apical cortex. RhoGEF2 pulses precede myosin, and C-GAP is required for pulsation, suggesting that contractile pulses result from RhoA activity cycling. Finally, C-GAP expression level influences the transition from reversible to irreversible cell shape change, which defines the onset of tissue shape change. Our data demonstrate that RhoA activity cycling and modulating the ratio of RhoGEF2 to C-GAP are required for tissue folding. PMID:27551058

  17. Incidence of apical crack initiation and propagation during the removal of root canal filling material with ProTaper and Mtwo rotary nickel-titanium retreatment instruments and hand files.

    PubMed

    Topçuoğlu, Hüseyin Sinan; Düzgün, Salih; Kesim, Bertan; Tuncay, Oznur

    2014-07-01

    The aim of this study was to determine the incidence of crack initiation and propagation in apical root dentin after retreatment procedures performed by using 2 rotary retreatment systems and hand files with additional instrumentation. Eighty extracted mandibular premolars with single canals were selected. One millimeter from the apex of each tooth was ground perpendicular to the long axis of the tooth, and the apical surface was polished. Twenty teeth served as the control group, and no preparation was performed. The remaining 60 teeth were prepared to size 35 with rotary files and filled with gutta-percha and AH Plus sealer. Specimens were then divided into 3 groups (n = 20), and retreatment procedures were performed with the following devices and techniques: ProTaper Universal retreatment files, Mtwo retreatment files, and hand files. After retreatment, the additional instrumentation was performed by using size 40 ProTaper, Mtwo, and hand files. Digital images of the apical root surface were recorded before preparation, after instrumentation, after filling, after retreatment, and after additional instrumentation. The images were then inspected for the presence of any new apical cracks and propagation. Data were analyzed with the logistic regression and Fisher exact tests. All experimental groups caused crack initiation and propagation after use of retreatment instruments. The ProTaper and Mtwo retreatment groups caused greater crack initiation and propagation than the hand instrument group (P < .05) after retreatment. Additional instrumentation with ProTaper and Mtwo instruments after the use of retreatment instruments caused crack initiation and propagation, whereas hand files caused neither crack initiation nor propagation (P < .05). This study showed that retreatment procedures and additional instrumentation after the use of retreatment files may cause crack initiation and propagation in apical dentin. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  18. Apical Extrusion of Intracanal Debris Using Two Engine Driven and Step-Back Instrumentation Techniques: An In-Vitro Study

    PubMed Central

    Kustarci, Alper; Akdemir, Neslihan; Siso, Seyda Herguner; Altunbas, Demet

    2008-01-01

    Objectives The purpose of this study was to compare in-vitro the amount of debris extruded apically from extracted teeth, using K3, Protaper rotary instruments and manual step-back technique. Methods Forty five human single-rooted mandibular premolar teeth were randomly divided into 3 groups. The teeth in 3 groups were instrumented until reaching the working length with K3, Protaper rotary instruments and K-type stainless steel instruments with manual step-back technique, respectively. Debris extruded from the apical foramen was collected into centrifuge tubes and the amount was determined. The data obtained were analyzed using Kruskal-Wallis one-way analysis of variance and Mann-Whitney U tests, with P=.05 as the level for statistical significance. Results Statistically significant difference was observed between K3, Protaper and step-back groups in terms of debris extrusion (P<.05). Step-back group had the highest mean debris weight, which was significantly different from the K3 and Protaper groups (P<.05). The lowest mean debris weight was related to K3 group, which was significantly different from the Protaper group (P<.05). Conclusions: Based on the results, all instrumentation techniques produced debris extrusion. The engine-driven Ni-Ti systems extruded significantly less apical debris than step-back technique. However, Protaper rotary instruments extruded significantly more debris than K3 rotary instruments. PMID:19212528

  19. Apical extrusion of intracanal debris using two engine driven and step-back instrumentation techniques: an in-vitro study.

    PubMed

    Kustarci, Alper; Akdemir, Neslihan; Siso, Seyda Herguner; Altunbas, Demet

    2008-10-01

    The purpose of this study was to compare in-vitro the amount of debris extruded apically from extracted teeth, using K3, Protaper rotary instruments and manual step-back technique. Forty five human single-rooted mandibular premolar teeth were randomly divided into 3 groups. The teeth in 3 groups were instrumented until reaching the working length with K3, Protaper rotary instruments and K-type stainless steel instruments with manual step-back technique, respectively. Debris extruded from the apical foramen was collected into centrifuge tubes and the amount was determined. The data obtained were analyzed using Kruskal-Wallis one-way analysis of variance and Mann-Whitney U tests, with P=.05 as the level for statistical significance. Statistically significant difference was observed between K3, Protaper and step-back groups in terms of debris extrusion (P<.05). Step-back group had the highest mean debris weight, which was significantly different from the K3 and Protaper groups (P<.05). The lowest mean debris weight was related to K3 group, which was significantly different from the Protaper group (P<.05). Based on the results, all instrumentation techniques produced debris extrusion. The engine-driven Ni-Ti systems extruded significantly less apical debris than step-back technique. However, Protaper rotary instruments extruded significantly more debris than K3 rotary instruments.

  20. Expression of apical Na(+)-L-glutamine co-transport activity, B(0)-system neutral amino acid co-transporter (B(0)AT1) and angiotensin-converting enzyme 2 along the jejunal crypt-villus axis in young pigs fed a liquid formula

    USDA-ARS?s Scientific Manuscript database

    Gut apical amino acid (AA) transport activity is high at birth and during suckling, thus being essential to maintain luminal nutrient-dependent mucosal growth through providing AA as essential metabolic fuel, substrates and nutrient stimuli for cellular growth. Because system-B(0) Na(+)-neutral AA c...

  1. The Spectrin cytoskeleton regulates the Hippo signalling pathway.

    PubMed

    Fletcher, Georgina C; Elbediwy, Ahmed; Khanal, Ichha; Ribeiro, Paulo S; Tapon, Nic; Thompson, Barry J

    2015-04-01

    The Spectrin cytoskeleton is known to be polarised in epithelial cells, yet its role remains poorly understood. Here, we show that the Spectrin cytoskeleton controls Hippo signalling. In the developing Drosophila wing and eye, loss of apical Spectrins (alpha/beta-heavy dimers) produces tissue overgrowth and mis-regulation of Hippo target genes, similar to loss of Crumbs (Crb) or the FERM-domain protein Expanded (Ex). Apical beta-heavy Spectrin binds to Ex and co-localises with it at the apical membrane to antagonise Yki activity. Interestingly, in both the ovarian follicular epithelium and intestinal epithelium of Drosophila, apical Spectrins and Crb are dispensable for repression of Yki, while basolateral Spectrins (alpha/beta dimers) are essential. Finally, the Spectrin cytoskeleton is required to regulate the localisation of the Hippo pathway effector YAP in response to cell density human epithelial cells. Our findings identify both apical and basolateral Spectrins as regulators of Hippo signalling and suggest Spectrins as potential mechanosensors. © 2015 The Authors. Published under the terms of the CC BY 4.0 license.

  2. Sealing ability of mineral trioxide aggregate (MTA) combined with distilled water, chlorhexidine, and doxycycline.

    PubMed

    Arruda, Roberta A A; Cunha, Rodrigo S; Miguita, Kenner B; Silveira, Cláudia F M; De Martin, Alexandre S; Pinheiro, Sérgio L; Rocha, Daniel G P; Bueno, Carlos E S

    2012-09-01

    The aim of this study was to evaluate the sealing ability of mineral trioxide aggregate (MTA Bio) combined with different mixing agents (distilled water, chlorhexidine, doxycycline), used as an apical root-end filling material. Forty-two extracted human teeth were divided into three groups (n = 12); six teeth were used as controls. Root-ends were resected at 90 degrees, 3 mm from the apex. Root-end cavities were prepared using ultrasonic tips and filled with MTA Bio plus distilled water, 2% chlorhexidine solution, or 10% doxycycline solution. Apical sealing was assessed by microleakage of 50% silver nitrate solution. Roots were longitudinally sectioned in a buccolingual plane and analyzed using an operating microscope (20× magnification). Depth of dye leakage into the dentinal walls was measured in millimeters. Results were analyzed using ANOVA and Tukey's test (P = 0.05). MTA Bio plus distilled water showed significantly higher mean leakage results (1.06 mm) when compared with MTA Bio plus doxycycline (0.61 mm), and higher, although not significant, results when compared with MTA Bio plus chlorhexidine (0.79 mm). In conclusion, replacing distilled water with two biologically active mixing agents (doxycycline and chlorhexidine) did not alter the sealing properties of MTABio. The antimicrobial properties of these combinations should be further investigated.

  3. Influence of Cone-beam Computed Tomography on Endodontic Retreatment Strategies among General Dental Practitioners and Endodontists.

    PubMed

    Rodríguez, Gustavo; Patel, Shanon; Durán-Sindreu, Fernando; Roig, Miguel; Abella, Francesc

    2017-09-01

    Treatment options for endodontic failure include nonsurgical or surgical endodontic retreatment, intentional replantation, and extraction with or without replacement of the tooth. The aim of the present study was to determine the impact of cone-beam computed tomographic (CBCT) imaging on clinical decision making among general dental practitioners and endodontists after failed root canal treatment. A second objective was to assess the self-reported level of difficulty in making a treatment choice before and after viewing a preoperative CBCT scan. Eight patients with endodontically treated teeth diagnosed as symptomatic apical periodontitis, acute apical abscess, or chronic apical abscess were selected. In the first session, the examiners were given the details of each case, including any relevant radiographs, and were asked to choose 1 of the proposed treatment alternatives and assess the difficulty of making a decision. One month later, the examiners reviewed randomly the same 8 cases with the additional information from the CBCT data. The examiners altered their treatment plan after viewing the CBCT scan in 49.8% of the cases. A significant difference in the treatment plan between the 2 imaging modalities was recorded for endodontists and general practitioners (P < .05). After CBCT evaluation, neither group altered their self-reported level of difficulty when choosing a treatment plan (P = .0524). The extraction option rose significantly to 20% after viewing the CBCT scan (P < .05). CBCT imaging directly influences endodontic retreatment strategies among general dental practitioners and endodontists. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  4. External apical root resorption in maxillary incisors in orthodontic patients: associated factors and radiographic evaluation.

    PubMed

    Nanekrungsan, Kamonporn; Patanaporn, Virush; Janhom, Apirum; Korwanich, Narumanus

    2012-09-01

    This study was performed to evaluate the incidence and degree of external apical root resorption of maxillary incisors after orthodontic treatment and to evaluate particular associated factors related to external apical root resorption. The records and maxillary incisor periapical radiographs of 181 patients were investigated. Crown and root lengths were measured and compared on the pre- and post-treatment periapical radiographs. Crown length was measured from the center of the incisal edge to the midpoint of the cemento-enamel junction (CEJ). Root length was measured from the CEJ midpoint to the root apex. A correction factor for the enlargement difference was used to calculate root resorption. The periapical radiographs of 564 teeth showed that the average root resorption was 1.39±1.27 (8.24±7.22%) and 1.69±1.14 mm (10.16±6.78%) for the maxillary central and lateral incisors, respectively. The results showed that the dilacerated or pointed roots, maxillary premolar extraction cases, and treatment duration were highly significant factors for root resorption (p<0.001). Allergic condition was a significant factor at p<0.01. Age at the start of treatment, large overjet, and history of facial trauma were also factors significantly associated with root resorption (p<0.05). There was no statistically significant difference in root resorption among the factors of gender, overbite, tongue-thrusting habit, types of malocclusion, and types of bracket. These results suggested that orthodontic treatment should be carefully performed in pre-treatment extraction patients who have pointed or dilacerated roots and need long treatment duration.

  5. Effect of phospholipid-based formulations of Boswellia serrata extract on the solubility, permeability, and absorption of the individual boswellic acid constituents present.

    PubMed

    Hüsch, Jan; Gerbeth, Kathleen; Fricker, Gert; Setzer, Constanze; Zirkel, Jürgen; Rebmann, Herbert; Schubert-Zsilavecz, Manfred; Abdel-Tawab, Mona

    2012-10-26

    Boswellia serrata gum resin extracts are used widely for the treatment of inflammatory diseases. However, very low concentrations in the plasma and brain were observed for the boswellic acids (1-6, the active constituents of B. serrata). The present study investigated the effect of phospholipids alone and in combination with common co-surfactants (e.g., Tween 80, vitamin E-TPGS, pluronic f127) on the solubility of 1-6 in physiologically relevant media and on the permeability in the Caco-2 cell model. Because of the high lipophilicity of 1-6, the permeability experiments were adapted to physiological conditions using modified fasted state simulated intestinal fluid as apical (donor) medium and 4% bovine serum albumin in the basolateral (receiver) compartment. A formulation composed of extract/phospholipid/pluronic f127 (1:1:1 w/w/w) increased the solubility of 1-6 up to 54 times compared with the nonformulated extract and exhibited the highest mass net flux in the permeability tests. The oral administration of this formulation to rats (240 mg/kg) resulted in 26 and 14 times higher plasma levels for 11-keto-β-boswellic acid (1) and acetyl-11-keto-β-boswellic acid (2), respectively. In the brain, five times higher levels for 2 compared to the nonformulated extract were determined 8 h after oral administration.

  6. Bicarbonate, NBCe1, NHE, and Carbonic Anhydrase Activity Enhance Lactate-H+ Transport in Bovine Corneal Endothelium

    PubMed Central

    Nguyen, Tracy T.

    2011-01-01

    Purpose. To identify and localize the monocarboxylate transporters (MCTs) expressed in bovine corneal endothelial cells (BCEC) and to test the hypothesis that buffering contributed by HCO3−, sodium bicarbonate cotransporter (NBCe1), sodium hydrogen exchanger (NHE), and carbonic anhydrase (CA) activity facilitates lactate flux. Methods. MCT1–4 expression was screened by RT-PCR, Western blot analysis, and immunofluorescence. Endogenous lactate efflux and/or pHi were measured in BCEC in HCO3−-free or HCO3−-rich Ringer, with and without niflumic acid (MCT inhibitor), acetazolamide (ACTZ, a CA inhibitor), 5-(N-Ethyl-N-isopropyl)amiloride (EIPA) (Na+/H+ exchange blocker), disodium 4,4′-diisothiocyanatostilbene-2,2′-disulfonate (DIDS; anion transport inhibitor), or with NBCe1-specific small interfering (si) RNA-treated cells. Results. MCT1, 2, and 4 are expressed in BCEC. MCT1 was localized to the lateral membrane, MCT2 was lateral and apical, while MCT4 was apical. pHi measurements showed significant lactate-induced cell acidification (LIA) in response to 20-second pulses of lactate. Incubation with niflumic acid significantly reduced the rate of pHi change (dpHi/dt) and lactate-induced cell acidification. EIPA inhibited alkalinization after lactate removal. Lactate-dependent proton flux was significantly greater in the presence of HCO3− but was reduced by ACTZ. Efflux of endogenously produced lactate was significantly faster in the presence of HCO3−, was greater on the apical surface, was reduced on the apical side by ACTZ, as well as on the apical and basolateral side by NBCe1-specific siRNA, DIDS, or EIPA. Conclusions. MCT1, 2, and 4 are expressed in BCEC on both the apical and basolateral membrane (BL) surfaces consistent with niflumic acid-sensitive lactate-H+ transport. Lactate dependent proton flux can activate Na+/H+ exchange and be facilitated by maximizing intracellular buffering capacity through the presence of HCO3−, HCO3− transport, NHE and CA activity. PMID:21896839

  7. MST4 kinase phosphorylates ACAP4 protein to orchestrate apical membrane remodeling during gastric acid secretion.

    PubMed

    Yuan, Xiao; Yao, Phil Y; Jiang, Jiying; Zhang, Yin; Su, Zeqi; Yao, Wendy; Wang, Xueying; Gui, Ping; Mullen, McKay; Henry, Calmour; Ward, Tarsha; Wang, Wenwen; Brako, Larry; Tian, Ruijun; Zhao, Xuannv; Wang, Fengsong; Cao, Xinwang; Wang, Dongmei; Liu, Xing; Ding, Xia; Yao, Xuebiao

    2017-09-29

    Digestion in the stomach depends on acidification of the lumen. Histamine-elicited acid secretion is triggered by activation of the PKA cascade, which ultimately results in the insertion of gastric H,K-ATPases into the apical plasma membranes of parietal cells. Our recent study revealed the functional role of PKA-MST4-ezrin signaling axis in histamine-elicited acid secretion. However, it remains uncharacterized how the PKA-MST4-ezrin signaling axis operates the insertion of H,K-ATPases into the apical plasma membranes of gastric parietal cells. Here we show that MST4 phosphorylates ACAP4, an ARF6 GTPase-activating protein, at Thr 545 Histamine stimulation activates MST4 and promotes MST4 interaction with ACAP4. ACAP4 physically interacts with MST4 and is a cognate substrate of MST4 during parietal cell activation. The phosphorylation site of ACAP4 by MST4 was mapped to Thr 545 by mass spectrometric analyses. Importantly, phosphorylation of Thr 545 is essential for acid secretion in parietal cells because either suppression of ACAP4 or overexpression of non-phosphorylatable ACAP4 prevents the apical membrane reorganization and proton pump translocation elicited by histamine stimulation. In addition, persistent overexpression of MST4 phosphorylation-deficient ACAP4 results in inhibition of gastric acid secretion and blockage of tubulovesicle fusion to the apical membranes. Significantly, phosphorylation of Thr 545 enables ACAP4 to interact with ezrin. Given the location of Thr 545 between the GTPase-activating protein domain and the first ankyrin repeat, we reason that MST4 phosphorylation elicits a conformational change that enables ezrin-ACAP4 interaction. Taken together, these results define a novel molecular mechanism linking the PKA-MST4-ACAP4 signaling cascade to polarized acid secretion in gastric parietal cells. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  8. Rhubarb Tannins Extract Inhibits the Expression of Aquaporins 2 and 3 in Magnesium Sulphate-Induced Diarrhoea Model

    PubMed Central

    Liu, Chunfang; Zheng, Yanfang; Xu, Wen; Wang, Hui

    2014-01-01

    Tannins, a group of major active components of Chinese rhubarb and widely distributed in nature, have a significant antidiarrhoeal activity. Aquaporins (AQPs) 2 and 3 play important roles in regulating water transfer during diarrhoea. The present study aims to determine the effect of the total tannins extract of rhubarb on aquaporins (AQPs) 2 and 3 in diarrhoea mice and HT-29 cells both induced by magnesium sulphate (MgSO4). Our results showed that rhubarb tannins extract (RTE) significantly decreased the faecal water content in colon and evaluation index of defecation of diarrhoea mice. Interestingly, RTE could markedly reduce the mRNA and protein expression levels of AQPs 2 and 3 in apical and lateral mucosal epithelial cells in the colons of diarrhoea mice and HT-29 cells both induced by MgSO4 in a dose-dependent manner. Furthermore, RTE suppressed the production of cyclic monophosphate- (cAMP-) dependent protein kinase A catalytic subunits α (PKA C-α) and phosphorylated cAMP response element-binding protein (p-CREB, Ser133) in MgSO4-induced HT-29 cells. Our data showed for the first time that RTE inhibit AQPs 2 and 3 expression in vivo and in vitro via downregulating PKA/p-CREB signal pathway, which accounts for the antidiarrhoeal effect of RTE. PMID:25215286

  9. Distribution of Porphyromonas gingivalis fimA genotypes in chronic apical periodontitis associated with symptoms.

    PubMed

    Wang, Qian; Zhou, Xue-dong; Zheng, Qing-hua; Wang, Yao; Tang, Lu; Huang, Ding-ming

    2010-11-01

    Porphyromonas gingivalis (P. gingivalis) is an anaerobic bacterium involved in root canal infections whose fimbriae are classified into six genotypes (types I-V and Ib) based on nucleotide sequence. Accumulated evidence suggests there is significant association between P. gingivalis and some clinical symptoms of periodontal diseases. The present study aims to determine the prevalence of P. gingivalis fimA genotypes in apical periodontitis and to investigate the correlation between P. gingivalis fimA genotypes and clinical symptoms. Samples were obtained from 158 infected root canals with apical periodontitis. DNA was extracted and analyzed with a polymerase chain reaction-based identification assay. Odds ratios, 95% confidence intervals, and contingency coefficient were calculated for associating the fimA-specific genes with clinical symptoms. P. gingivalis was detected in 39.9% of the inflected root canal samples and was found in 44.5% of P. gingivalis-positive specimens with symptoms. Types II (69.4%) were the most frequent in the symptomatic cases followed by type IV (32.7%). The occurrence of type I (64.3%) was significantly higher than any other genotypes in the asymptomatic apical periodontitis, whereas type II and type Ib were not identified. Statistical analysis revealed that the occurrences of types II, IV, and Ib fimA were associated with greater risk of clinical signs (swelling, sinus tract, or intracanal exudates) than type I. Results from this study reinforce the association between P. gingivalis-specific fimA genotypic clones and apical periodontitis, indicating that fimA genotypes (types II, IV, and Ib) were related to the etiology of symptomatic periradicular diseases. Copyright © 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  10. Analysis of symptomatic and asymptomatic primary root canal infections in adult Norwegian patients.

    PubMed

    Rôças, Isabela N; Siqueira, José F; Debelian, Gilberto J

    2011-09-01

    This molecular study analyzed the microbiota of primary root canal infections from adult Norwegian patients. Samples were taken from the necrotic root canals of teeth with symptomatic (n = 13) or asymptomatic (n = 21) apical periodontitis and chronic apical abscesses (n = 9). DNA was extracted from samples, and bacterial identifications were performed by a closed-ended reverse-capture checkerboard approach targeting 50 candidate endodontic pathogens. Bacterial DNA was detected in all cases. In teeth with asymptomatic apical periodontitis, the most frequent taxa were Dialister invisus (71%), Fusobacterium nucleatum (62%), and Porphyromonas endodontalis (62%). In chronic apical abscesses, the most prevalent taxa were P. endodontalis (100%), D. invisus (89%), Parvimonas micra (78%), and Solobacterium moorei (78%). In teeth with symptomatic apical periodontitis, the most prevalent taxa were D. invisus, P. endodontalis, S. moorei, Propionibacterium acnes, and Streptococcus species (all in 69%). None of the targeted taxa were significantly associated with either sinus tract or pain (P > .05), except for Selenomonas sputigena, which was more frequently found in painful cases (P = .04). No taxa were found in significantly higher levels in any conditions (P > .05). Cluster analyses revealed bacterial groupings that differed between cases with and without pain. Although basically the same species were highly prevalent in the different conditions examined and none of the most prevalent taxa were positively associated with symptoms, results revealed that species formed different partnerships and associations in samples from teeth with or without pain. Therefore, it is possible that more virulent multispecies communities can form as a result of overall bacterial combinations and give rise to acute inflammation. Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  11. Viral-bacterial associations in acute apical abscesses.

    PubMed

    Ferreira, Dennis C; Rôças, Isabela N; Paiva, Simone S M; Carmo, Flávia L; Cavalcante, Fernanda S; Rosado, Alexandre S; Santos, Kátia R N; Siqueira, José F

    2011-08-01

    Viral-bacterial and bacterial synergism have been suggested to contribute to the pathogenesis of several human diseases. This study sought to investigate the possible associations between 9 candidate endodontic bacterial pathogens and 9 human viruses in samples from acute apical abscesses. DNA extracts from purulent exudate aspirates of 33 cases of acute apical abscess were surveyed for the presence of 9 selected bacterial species using a 16S ribosomal RNA gene-based nested polymerase chain reaction (PCR) approach. Single or nested PCR assays were used for detection of the human papillomavirus (HPV) and herpesviruses types 1 to 8. Two-thirds of the abscess samples were positive for at least one of the target viruses. Specifically, the most frequently detected viruses were HHV-8 (54.5%); HPV (9%); and varicella zoster virus (VZV), Epstein-Barr virus (EBV), and HHV-6 (6%). Bacterial DNA was present in all cases and the most prevalent bacterial species were Treponema denticola (70%), Tannerella forsythia (67%), Porphyromonas endodontalis (67%), Dialister invisus (61%), and Dialister pneumosintes (57.5%). HHV-8 was positively associated with 7 of the target bacterial species and HPV with 4, but all these associations were weak. Several bacterial pairs showed a moderate positive association. Viral coinfection was found in 6 abscess cases, but no significant viral association could be determined. Findings demonstrated that bacterial and viral DNA occurred concomitantly in two-thirds of the samples from endodontic abscesses. Although this may suggest a role for viruses in the etiology of apical abscesses, the possibility also exists that the presence of viruses in abscess samples is merely a consequence of the bacterially induced disease process. Further studies are necessary to clarify the role of these viral-bacterial interactions, if any, in the pathogenesis of acute apical abscesses. Copyright © 2011 Mosby, Inc. All rights reserved.

  12. Incidence of Apical Crack Initiation during Canal Preparation using Hand Stainless Steel (K-File) and Hand NiTi (Protaper) Files

    PubMed Central

    Raisingani, Deepak; Mathur, Rachit; Madan, Nidha; Visnoi, Suchita

    2016-01-01

    Aim To evaluate the incidence of apical crack initiation during canal preparation with stainless steel K-files and hand protaper files (in vitro study). Materials and methods Sixty extracted mandibular premo-lar teeth are randomly selected and embedded in an acrylic tube filled with autopolymerizing resin. A baseline image of the apical surface of each specimen was recorded under a digital microscope (80×). The cervical and middle thirds of all samples were flared with #2 and #1 Gates-Glidden (GG) drills, and a second image was recorded. The teeth were randomly divided into four groups of 15 teeth each according to the file type (hand K-file and hand-protaper) and working length (WL) (instrumented at WL and 1 mm less than WL). Final image after dye penetration and photomicrograph of the apical root surface were digitally recorded. Results Maximum numbers of cracks were observed with hand protaper files compared with hand K-file at the WL and 1 mm short of WL. Chi-square testing revealed a highly significant effect of WL on crack formation at WL and 1 mm short of WL (p = 0.000). Conclusion Minimum numbers of cracks at WL and 1 mm short of WL were observed with hand K-file and maximum with hand protaper files. How to cite this article Soni D, Raisingani D, Mathur R, Madan N, Visnoi S. Incidence of Apical Crack Initiation during Canal Preparation using Hand Stainless Steel (K-File) and Hand NiTi (Protaper) Files. Int J Clin Pediatr Dent 2016;9(4):303-307. PMID:28127160

  13. Effect of Calcium Hydroxide, Chlorhexidine Digluconate and Camphorated Monochlorophenol on the Sealing Ability of Biodentine Apical Plug.

    PubMed

    Srivastava, Aastha Arora; Srivastava, Harshit; Prasad, Ashwini B; Raisingani, Deepak; Soni, Dileep

    2016-06-01

    Teeth with immature apex are managed by establishing an apical plug using various materials and techniques. However, the use of previously placed intracanal medicament may affect the sealing ability of permanent filling material used as an apical plug. To evaluate the effect of removal of previously placed Calcium Hydroxide, Chlorhexidine Digluconate and Camphorated Monochlorophenol as an intracanal medicament on the sealing ability of the Biodentine as an apical plug. A total of 72 recently extracted human permanent teeth with single root were selected and stored in saline at room temperature. The crown portion of each tooth was removed at the level of cemento enamel junction; 14mm root length was taken as standard length. All the roots were submerged in 20% sulphuric acid up to 3 mm from the apex, for four days for root resorption. One sample was cut longitudinally to look for root resorption under stereo microscope. The canal preparation was done; the roots were kept in moist gauze after instrumentation. A total of 71 roots were randomly divided into three groups. GROUP 1:Calcium hydroxide paste, GROUP 2: Chlorhexidine digluconate, GROUP 3: Camphorated Monochlorophenol (CMCP). The medicaments were removed with stainless steel hand files and 0.5% sodium hypochlorite irrigation. After removal of medicament Biodentine was placed in apical third of resorbed roots and the remaining portion of the canals was filled with gutta-percha. All the 71 roots were analysed with fluid filtration method for evaluating microleakage. Comparing all the three groups statistically there was no significant difference. The mean values were found more for group 1 followed by group 2 & 3. All the groups showed microleakage. Calcium hydroxide showed the maximum microleakage followed by Chlorhexidine digluconate and least with CMCP.

  14. ELMOD1 Stimulates ARF6-GTP Hydrolysis to Stabilize Apical Structures in Developing Vestibular Hair Cells.

    PubMed

    Krey, Jocelyn F; Dumont, Rachel A; Wilmarth, Philip A; David, Larry L; Johnson, Kenneth R; Barr-Gillespie, Peter G

    2018-01-24

    Sensory hair cells require control of physical properties of their apical plasma membranes for normal development and function. Members of the ADP-ribosylation factor (ARF) small GTPase family regulate membrane trafficking and cytoskeletal assembly in many cells. We identified ELMO domain-containing protein 1 (ELMOD1), a guanine nucleoside triphosphatase activating protein (GAP) for ARF6, as the most highly enriched ARF regulator in hair cells. To characterize ELMOD1 control of trafficking, we analyzed mice of both sexes from a strain lacking functional ELMOD1 [roundabout ( rda )]. In rda/rda mice, cuticular plates of utricle hair cells initially formed normally, then degenerated after postnatal day 5; large numbers of vesicles invaded the compromised cuticular plate. Hair bundles initially developed normally, but the cell's apical membrane lifted away from the cuticular plate, and stereocilia elongated and fused. Membrane trafficking in type I hair cells, measured by FM1-43 dye labeling, was altered in rda/rda mice. Consistent with the proposed GAP role for ELMOD1, the ARF6 GTP/GDP ratio was significantly elevated in rda/rda utricles compared with controls, and the level of ARF6-GTP was correlated with the severity of the rda/rda phenotype. These results suggest that conversion of ARF6 to its GDP-bound form is necessary for final stabilization of the hair bundle. SIGNIFICANCE STATEMENT Assembly of the mechanically sensitive hair bundle of sensory hair cells requires growth and reorganization of apical actin and membrane structures. Hair bundles and apical membranes in mice with mutations in the Elmod1 gene degenerate after formation, suggesting that the ELMOD1 protein stabilizes these structures. We show that ELMOD1 is a GTPase-activating protein in hair cells for the small GTP-binding protein ARF6, known to participate in actin assembly and membrane trafficking. We propose that conversion of ARF6 into the GDP-bound form in the apical domain of hair cells is essential for stabilizing apical actin structures like the hair bundle and ensuring that the apical membrane forms appropriately around the stereocilia. Copyright © 2018 the authors 0270-6474/18/380843-15$15.00/0.

  15. Proteomic Profiling and Differential Messenger RNA Expression Correlate HSP27 and Serpin Family B Member 1 to Apical Periodontitis Outcomes.

    PubMed

    Cavalla, Franco; Biguetti, Claudia; Jain, Sameer; Johnson, Cleverick; Letra, Ariadne; Garlet, Gustavo Pompermaier; Silva, Renato Menezes

    2017-09-01

    Understanding protein expression profiles of apical periodontitis may contribute to the discovery of novel diagnostic or therapeutic molecular targets. Periapical tissue samples (n = 5) of patients with lesions characterized as nonhealing were submitted for proteomic analysis. Two differentially expressed proteins (heat shock protein 27 [HSP27] and serpin family B member 1 [SERPINB1]) were selected for characterization, localization by immunofluorescence, and association with known biomarkers of acute inflammatory response in human apical periodontitis (n = 110) and healthy periodontal ligaments (n = 26). Apical periodontitis samples were categorized as stable/inactive (n = 70) or progressive/active (n = 40) based on the ratio of expression of receptor activator of nuclear factor kappa-B ligand (RANKL)/osteoprotegerin (OPG). Next, the expression of HSP27, SERPINB1, C-X-C motif Chemokine Receptor 1 (CXCR1), matrix metalloproteinase 8 (MMP8), myeloperoxidase (MPO), and cathepsin G (CTSG) messenger RNA was evaluated using real-time polymerase chain reaction. Data analysis was performed using the Shapiro-Wilk test, analysis of variance, and the Pearson test. P values <.05 were considered statistically significant. Proteomic analysis revealed 48 proteins as differentially expressed in apical periodontitis compared with a healthy periodontium, with 30 of these proteins found to be expressed in all 4 lesions. The expression of HSP27 and SERPINB1 was ∼2-fold higher in apical periodontitis. Next, an increased expression of HSP27 was detected in epithelial cells, whereas SERPINB1 expression was noted in neutrophils and epithelial cells. HSP27 and SERPINB1 transcripts were highly expressed in stable/inactive lesions (P < .05). Significant negative correlations were found between the expression of HSP27 and SERPINB1 with biomarkers of acute inflammation including CXCR1, MPO, and CTSG. Our data suggest HSP27 and SERPINB1 as potential regulators of the inflammatory response in apical periodontitis. Additional functional studies should be performed to further characterize the role of these molecules during the development/progression of apical periodontitis. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  16. Ginger Extract Suppresses Inflammatory Response and Maintains Barrier Function in Human Colonic Epithelial Caco-2 Cells Exposed to Inflammatory Mediators.

    PubMed

    Kim, Yunyoung; Kim, Dong-Min; Kim, Ji Yeon

    2017-05-01

    The beneficial effects of ginger in the management of gastrointestinal disturbances have been reported. In this study, the anti-inflammatory potential of ginger extract was assessed in a cellular model of gut inflammation. In addition, the effects of ginger extract and its major active compounds on intestinal barrier function were evaluated. The response of Caco-2 cells following exposure to a mixture of inflammatory mediators [interleukin [IL]-1β, 25 ng/mL; lipopolysaccharides [LPS], 10 ng/mL; tumor necrosis factor [TNF]-α, 50 ng/mL; and interferon [INF]-γ, 50 ng/mL] were assessed by measuring the levels of secreted IL-6 and IL-8. In addition, the mRNA levels of cyclooxygenase-2 and inducible nitric oxide synthase were measured. Moreover, the degree of nuclear factor (NF)-κB inhibition was examined, and the intestinal barrier function was determined by measuring the transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC)-dextran transfer. It was observed that ginger extract and its constituents improved inflammatory responses by decreasing the levels of nitrite, PGE2, IL-6, and IL-8 via NF-κB inhibition. The ginger extract also increased the TEER and decreased the transfer of FITC-dextran from the apical side of the epithelium to the basolateral side. Taken together, these results show that ginger extract may be developed as a functional food for the maintenance of gastrointestinal health. © 2017 Institute of Food Technologists®.

  17. An in vitro and in silico study on the flavonoid-mediated modulation of the transport of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) through Caco-2 monolayers

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Schutte, Maaike E.; Freidig, Andreas P.; Sandt, Johannes J.M. van de

    The present study describes the effect of different flavonoids on the absorption of the pro-carcinogen PhIP through Caco-2 monolayers and the development of an in silico model describing this process taking into account passive diffusion and active transport of PhIP. Various flavonoids stimulated the apical to basolateral PhIP transport. Using the in silico model for flavone, kaempferol and chrysoeriol, the apparent Ki value for inhibition of the active transport to the apical side was estimated to be below 53 {mu}M and for morin, robinetin and taxifolin between 164 and 268 {mu}M. For myricetin, luteolin, naringenin and quercetin, the apparent Kimore » values were determined more accurately and amounted to 37.3, 12.2, 11.7 and 5.6 {mu}M respectively. Additional experiments revealed that the apical to basolateral PhIP transport was also increased in the presence of a typical BCRP or MRP inhibitor with apparent Ki values in the same range as those of the flavonoids. This observation together with the fact that flavonoids are known to be inhibitors of MRPs and BCRP, corroborates that inhibition of these apical membrane transporters is involved in the flavonoid-mediated increased apical to basolateral PhIP transport. Based on the apparent Ki values obtained, it is concluded that the flavonols, at the levels present in the regular Western diet, are capable of stimulating the transport of PhIP through Caco-2 monolayers from the apical to the basolateral compartment. This points to flavonoid-mediated stimulation of the bioavailability of PhIP and, thus, a possible adverse effect of these supposed beneficial food ingredients.« less

  18. Sequential activation of apical and basolateral contractility drives ascidian endoderm invagination

    PubMed Central

    Sherrard, Kristin; Robin, François; Lemaire, Patrick; Munro1, Edwin

    2014-01-01

    SUMMARY Background Epithelial invagination is a fundamental morphogenetic behavior that transforms a flat cell sheet into a pit or groove. Previous studies of invagination have focused on the role of actomyosin-dependent apical contraction; other mechanisms remain largely unexplored. Results We combined experimental and computational approaches to identify a two-step mechanism for endoderm invagination during ascidian gastrulation. During Step 1, which immediately precedes invagination, endoderm cells constrict their apices due to Rho/Rhokinase-dependent apical enrichment of 1P–myosin. Our data suggest that endoderm invagination itself occurs during Step 2, without further apical shrinkage, via a novel mechanism we call collared rounding: Rho/Rho-kinase-independent lateral enrichment of 1P–myosin drives apico-basal shortening, while Rho/Rho-kinase-dependent enrichment of 1P and 2P myosin in circumapical collars is required to prevent apical expansion and for deep invagination. Simulations show that boundary-specific tension values consistent with these distributions of active myosin can explain the cell shape changes observed during invagination both in normal embryos and in embryos treated with pharmacological inhibitors of either Rho-kinase or Myosin II ATPase. Indeed, we find that the balance of strong circumapical and basolateral tension is the only mechanism based on differential cortical tension that can explain ascidian endoderm invagination. Finally, simulations suggest that mesectoderm cells resist endoderm shape changes during both steps and we confirm this prediction experimentally. Conclusions Our findings suggest that early ascidian gastrulation is driven by the coordinated apposition of circumapical and lateral endoderm contraction, working against a resisting mesectoderm. We propose that similar mechanisms may operate during other invaginations. PMID:20691592

  19. Inhibition of RHO-ROCK signaling enhances ICM and suppresses TE characteristics through activation of Hippo signaling in the mouse blastocyst

    PubMed Central

    Kono, Kanako; Tamashiro, Dana Ann A.; Alarcon, Vernadeth B.

    2014-01-01

    Specification of the trophectoderm (TE) and inner cell mass (ICM) lineages in the mouse blastocyst correlates with cell position, as TE derives from outer cells whereas ICM from inner cells. Differences in position are reflected by cell polarization and Hippo signaling. Only in outer cells, the apical-basal cell polarity is established, and Hippo signaling is inhibited in such a manner that LATS1 and 2 (LATS1/2) kinases are prevented from phosphorylating YAP, a key transcriptional co-activator of the TE-specifying gene Cdx2. However, the molecular mechanisms that regulate these events are not fully understood. Here, we showed that inhibition of RHO-ROCK signaling enhances ICM and suppresses TE characteristics through activation of Hippo signaling and disruption of apical-basal polarity. Embryos treated with ROCK inhibitor Y-27632 exhibited elevated expression of ICM marker NANOG and reduced expression of CDX2 at the blastocyst stage. Y-27632-treated embryos failed to accumulate YAP in the nucleus, although it was rescued by concomitant inhibition of LATS1/2. Segregation between apical and basal polarity regulators, namely PARD6B, PRKCZ, SCRIB, and LLGL1, was dampened by Y-27632 treatment, whereas some of the polarization events at the late 8-cell stage such as compaction and apical localization of p-ERM and tyrosinated tubulin occurred normally. Similar abnormalities of Hippo signaling and apical-basal polarization were also observed in embryos that were treated with RHO GTPases inhibitor. These results suggest that RHO-ROCK signaling plays an essential role in regulating Hippo signaling and cell polarization to enable proper specification of the ICM and TE lineages. PMID:24997360

  20. Influence of the parameters of the Er:YAG laser on the apical sealing of apicectomized teeth.

    PubMed

    Marques, Aparecida Maria Cordeiro; Gerbi, Marleny Elizabeth M M; dos Santos, Jean Nunes; Noia, Manuela Pimentel; Oliveira, Priscila Chagas; Brugnera Junior, Aldo; Zanin, Fátima Antonia Aparecida; Pinheiro, Antonio Luiz Barbosa

    2011-07-01

    Failures in the sealing of the tooth apex have been considered to be responsible for most of the failures of apical surgeries. The Er:YAG laser has been proposed as an alternative for the use of rotator instruments in surgical endodontics due to its precision, lack of vibration, less post-operative discomfort, bacterial reduction, and less stress for patients and professionals. Following approval by the ethics committee, 12 extracted human canines without previous endodontic treatment with anatomically normal roots and free from apical lesions were washed in running tap water and disinfected. The teeth were sectioned axially at the crown-root junction and submitted to routine endodontic treatment. The apical limit was set at 1 mm before the apical foramen. The root canals were routinely filled with Gutta-Percha points and Sealer 26 and were randomly distributed into two groups (n = 6). In group I, apicectomy was performed with the Er:YAG laser (KAVO KEY Laser II®, Germany, λ = 2.940 nm, pulsed mode, 2051 tip, with air spray cooling, 250 mJ/15 Hz). Apical cut was performed of perpendicular mode 3 mm from the apical foramen. In group II, the same procedures and the same sequence as above was used, varying only the parameters of the Er:YAG laser (400 mJ/6 Hz). Sealing of the cervical end the apex was carried out with acrylic resin; the roots were covered by a layer of epoxy glue and two layers of nail polish. The specimens were divided into groups and fixed, by the cervical third, on wax. Impermeabilization of the residual root apical third was performed following the same procedures used in the cervical third but the residual apex was left free from the impermeabilization. After that, the roots were immersed in a 2% methylene blue solution and placed in a bacteriological oven for 48 h and then washed in running tap water for 2 h. The samples were sagittally split into two parts. The segments were visually observed and the one showing the greatest level of dye leakage was selected and kept in an individual container and coded accordingly. Apical staining was measured using a stereoscopic magnifying glass, a compass, and a caliper. The measurement was performed by three endodontists, previously calibrated, and unaware of the sample coding. The results showed that group I showed the greatest level of dye leakage. There was a significantly difference between the groups (p = 0.001). It is concluded that the apicectomies carried out with 400 mJ/6 Hz showed the smallest infiltration value.

  1. A PDZ-interacting domain in CFTR is an apical membrane polarization signal

    PubMed Central

    Moyer, Bryan D.; Denton, Jerod; Karlson, Katherine H.; Reynolds, Donna; Wang, Shusheng; Mickle, John E.; Milewski, Michal; Cutting, Garry R.; Guggino, William B.; Li, Min; Stanton, Bruce A.

    1999-01-01

    Polarization of the cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated chloride channel, to the apical plasma membrane of epithelial cells is critical for vectorial transport of chloride in a variety of epithelia, including the airway, pancreas, intestine, and kidney. However, the motifs that localize CFTR to the apical membrane are unknown. We report that the last 3 amino acids in the COOH-terminus of CFTR (T-R-L) comprise a PDZ-interacting domain that is required for the polarization of CFTR to the apical plasma membrane in human airway and kidney epithelial cells. In addition, the CFTR mutant, S1455X, which lacks the 26 COOH-terminal amino acids, including the PDZ-interacting domain, is mispolarized to the lateral membrane. We also demonstrate that CFTR binds to ezrin-radixin-moesin–binding phosphoprotein 50 (EBP50), an apical membrane PDZ domain–containing protein. We propose that COOH-terminal deletions of CFTR, which represent about 10% of CFTR mutations, result in defective vectorial chloride transport, partly by altering the polarized distribution of CFTR in epithelial cells. Moreover, our data demonstrate that PDZ-interacting domains and PDZ domain–containing proteins play a key role in the apical polarization of ion channels in epithelial cells. J. Clin. Invest. 104:1353–1361 (1999). PMID:10562297

  2. A Comparative Evaluation of Gutta Percha Removal and Extrusion of Apical Debris by Rotary and Hand Files

    PubMed Central

    Ebenezar, A.V. Rajesh; Kumar, Mohan; Sivakumar, A.

    2014-01-01

    Aim: The aim of this study was to evaluate the efficacy of Protaper retreatment files in comparison with RaCe, K3 and H-files for removal of gutta-percha and apically extruded debris using volumetric analysis. Materials and Methods: Forty extracted single rooted maxillary incisor teeth with straight canals and mature apices were selected for the study. After access cavity preparation, apical patency was confirmed with a size 10 K-file extending 1mm beyond the point at which it was first visible at the apical end. Working lengths were determined with the use of size 15 K-file. The canals were prepared in a step-back technique and the master apical file was size 30 for all teeth. 3% sodium hypochlorite was used as an irrigant after each instrumentation. Before final rinse, size 20 K-file was passed 1mm beyond the apex to remove any dentinal shaving plugs and maintain the apical patency. Then the canals were dried with paper points. The root canal was filled using standard gutta-percha points and zinc oxide eugenol sealer under lateral condensation technique. The teeth were then randomly divided into four groups of ten teeth each based on the instrument used for gutta percha removal. All the rotary instruments used in this study were rotated at 300rpm. The instruments used were: Group 1 – RaCe Files, Group 2 – ProTaper retreatment Files, Group 3 – K3 Files and Group 4 – H Files. The volume of the obturating material was calculated before and after removal using volumetric analysis with spiral CT. The removal efficacy with each instrument was calculated and statistically analysed. Results: The results of the study show that the ProTaper retreatment files (Group 2) (97.4%) showed the highest efficiency in the removal of obturating material, which was followed by RaCe (95.74%), K3 (92.86%) and H files (90.14%) with the efficiency in the decreasing order. Similarly the mean apical extrusion in H files (0.000 ± 0.002) was significantly lower than all the rotary instruments. However, the difference among the rotary files were not statistically significant (p>0.05). Conclusion: ProTaper retreatment files show significant difference over other groups in removal of obturating material and can be the system of choice in endodontic retreatment procedures. PMID:25584299

  3. Comparative evaluation of apical extrusion of debris and irrigant with three rotary instruments using crown down technique - An in vitro study.

    PubMed

    Jindal, Rahul; Singh, Smita; Gupta, Siddharth; Jindal, Punita

    2012-01-01

    The purpose of this study was to evaluate and compare the apical extrusion of debris and irrigant using various rotary instruments with crown down technique in the instrumentation of root canals. Thirty freshly extracted human permanent straight rooted mandibular premolars with minimum root curvature of 0-10 ° were divided in three groups with 10 teeth in each group. Each group was instrumented using one of the three rotary instrumentation systems: Rotary Hero shapers, Rotary ProTaper and Rotary Mtwo. One ml of sterile water was used as an irrigant after using each instrument. Debris extruded was collected in pre weighed glass vials and the extruded irrigant was measured quantitatively by Myers and Montgomery method and was later evaporated. The weight of the dry extruded debris was calculated by comparing the pre and post instrumentation weight of glass vials for each group. Statistical analysis was done by using by a Kruskal-Wallis One-way ANOVA test. Statistical analysis showed that all the rotary instruments used in this study caused apical extrusion of debris and irrigant. A Statistically significant difference was observed with Rotary ProTaper and Rotary Mtwo groups when compared with Rotary Hero shapers. But no significant difference was observed between Rotary ProTaper and Rotary Mtwo groups. After instrumentation with different rotary instruments, Hero shapers showed a less apical extrusion of debris and irrigant.

  4. Three-rooted premolar analyzed by high-resolution and cone beam CT.

    PubMed

    Marca, Caroline; Dummer, Paul M H; Bryant, Susan; Vier-Pelisser, Fabiana Vieira; Só, Marcus Vinicius Reis; Fontanella, Vania; Dutra, Vinicius D'avila; de Figueiredo, José Antonio Poli

    2013-07-01

    The aim of this study was to analyze the variations in canal and root cross-sectional area in three-rooted maxillary premolars between high-resolution computed tomography (μCT) and cone beam computed tomography (CBCT). Sixteen extracted maxillary premolars with three distinct roots and fully formed apices were scanned using μCT and CBCT. Photoshop CS software was used to measure root and canal cross-sectional areas at the most cervical and the most apical points of each root third in images obtained using the two tomographic computed (CT) techniques, and at 30 root sections equidistant from both root ends using μCT images. Canal and root areas were compared between each method using the Student t test for paired samples and 95 % confidence intervals. Images using μCT were sharper than those obtained using CBCT. There were statistically significant differences in mean area measurements of roots and canals between the μCT and CBCT techniques (P < 0.05). Root and canal areas had similar variations in cross-sectional μCT images and became proportionally smaller in a cervical to apical direction as the cementodentinal junction was approached, from where the area then increased apically. Although variation was similar in the roots and canals under study, CBCT produced poorer image details than μCT. Although CBCT is a strong diagnosis tool, it still needs improvement to provide accuracy in details of the root canal system, especially in cases with anatomical variations, such as the three-rooted maxillary premolars.

  5. Evaluation of penetration depth of 2% chlorhexidine digluconate into root dentinal tubules using confocal laser scanning microscope.

    PubMed

    Vadhana, Sekar; Latha, Jothi; Velmurugan, Natanasabapathy

    2015-05-01

    This study evaluated the penetration depth of 2% chlorhexidine digluconate (CHX) into root dentinal tubules and the influence of passive ultrasonic irrigation (PUI) using a confocal laser scanning microscope (CLSM). Twenty freshly extracted anterior teeth were decoronated and instrumented using Mtwo rotary files up to size 40, 4% taper. The samples were randomly divided into two groups (n = 10), that is, conventional syringe irrigation (CSI) and PUI. CHX was mixed with Rhodamine B dye and was used as the final irrigant. The teeth were sectioned at coronal, middle and apical levels and viewed under CLSM to record the penetration depth of CHX. The data were statistically analyzed using Kruskal-Wallis and Mann-Whitney U tests. The mean penetration depths of 2% CHX in coronal, middle and apical thirds were 138 µm, 80 µm and 44 µm in CSI group, respectively, whereas the mean penetration depths were 209 µm, 138 µm and 72 µm respectively in PUI group. Statistically significant difference was present between CSI group and PUI group at all three levels (p < 0.01 for coronal third and p < 0.001 for middle and apical thirds). On intragroup analysis, both groups showed statistically significant difference among three levels (p < 0.001). Penetration depth of 2% CHX into root dentinal tubules is deeper in coronal third when compared to middle and apical third. PUI aided in deeper penetration of 2% CHX into dentinal tubules when compared to conventional syringe irrigation at all three levels.

  6. Trypsin impaired epithelial barrier function and induced IL-8 secretion through basolateral PAR-2: a lesson from a stratified squamous epithelial model.

    PubMed

    Shan, Jing; Oshima, Tadayuki; Chen, Xin; Fukui, Hirokazu; Watari, Jiro; Miwa, Hiroto

    2012-11-15

    Immune-mediated injury by the protease-activated receptor-2-interleukin-8 (PAR-2-IL8) pathway may underlie the development of gastroesophageal reflux disease (GERD). However, the localization of PAR-2 and the mechanism of PAR-2 activation remain unclear. This study aimed to address these questions on an esophageal stratified squamous epithelial model and in the human esophageal mucosa of GERD patients. Normal human esophageal epithelial cells were cultured with the air-liquid interface system to establish the model. SLIGKV-NH2 (PAR-2 synthetic agonist), trypsin (PAR-2 natural activator), and weak acid (pH 4, 5, and 6) were added to either the apical or basolateral compartment to evaluate their effects on transepithelial electrical resistance (TEER) and IL-8 production. PAR-2 localization was examined both in the cell model and biopsies from GERD patients by immunohistochemistry. Apical trypsin stimulation induced IL-8 accompanied by decreased TEER in vitro, whereas the effective concentration from the basolateral side was 10 times lower. SLIGKV-NH2 from basolateral but not apical stimulation induced IL-8 production. Apical weak acid stimulation did not influence TEER or IL-8 production. Immunohistochemistry showed intense reactivity of PAR-2 in the basal and suprabasal layers after stimulation with trypsin. A similar PAR-2 reactivity that was mainly located at the basal and suprabasal layers was detected in GERD patients. In conclusion, the activation of the PAR-2-IL-8 pathway probably occurred at the basal and suprabasal layers, while the esophageal epithelial barrier may influence the activation of PAR-2. Under proton pump inhibitor therapy, refluxed trypsin may remain active and be a potential agent in the pathogenesis of refractory GERD.

  7. Light microscopic study of periapical lesions associated with asymptomatic apical periodontitis.

    PubMed

    Kabak, S L; Kabak, Y S; Anischenko, S L

    2005-04-01

    The purpose of the study has been to evaluate the degree of chronic inflammation in tissues surrounding the apex of the tooth root in patients with apical periodontitis in the remission phase. The material included 37 apical granulomas and radicular cysts obtained as a result of apiectomy, and 20 teeth which were removed together with the focus of the periapical inflammation. Routine histological techniques, as well as the immunofluorescent and immuno-chemical methods were used to examine the material. Despite the absence of clinical symptoms in 23 of 57 cases, the morphological signs of chronic inflammation were observed in the apical area of the tooth root. Morphological signs of viral invasion of epithelial and stromal cells in the radicular cyst wall were revealed in six cases. The presence of the virus of Herpes simplex I in epithelial cells (five cases) and adenoviral invasion (one case) was confirmed by immuno-fluorescent and immuno-chemical methods. Histological examination often reveals morphological signs of an active inflammatory process in the periapical tissues of patients treated during clinical remission. In our opinion, the presence of viruses in the epithelial cells of the radicular cyst may contribute to the persistence of the active stage of the inflammatory process.

  8. cAmp activation of apical membrane Cl(-) channels: theoretical considerations for impedance analysis.

    PubMed Central

    Păunescu, T G; Helman, S I

    2001-01-01

    Transepithelial electrical impedance analysis provides a sensitive method to evaluate the conductances and capacitances of apical and basolateral plasma membranes of epithelial cells. Impedance analysis is complicated, due not only to the anatomical arrangement of the cells and their paracellular shunt pathways, but also in particular to the existence of audio frequency-dependent capacitances or dispersions. In this paper we explore implications and consequences of anatomically related Maxwell-Wagner and Cole-Cole dielectric dispersions that impose limitations, approximations, and pitfalls of impedance analysis when tissues are studied under widely ranging spontaneous rates of transport, and in particular when apical membrane sodium and chloride channels are activated by adenosine 3',5'-cyclic monophosphate (cAMP) in A6 epithelia. We develop the thesis that capacitive relaxation processes of any origin lead not only to dependence on frequency of the impedance locus, but also to the appearance of depressed semicircles in Nyquist transepithelial impedance plots, regardless of the tightness or leakiness of the paracellular shunt pathways. Frequency dependence of capacitance precludes analysis of data in traditional ways, where capacitance is assumed constant, and is especially important when apical and/or basolateral membranes exhibit one or more dielectric dispersions. PMID:11463629

  9. Ultrasonic irrigation of a maxillary lateral incisor with perforation of the apical third of the root.

    PubMed

    Tsurumachi, Tamotsu; Takita, Toshiya; Hashimoto, Kazuhiro; Katoh, Takeshi; Ogiso, Bunnai

    2010-12-01

    We describe the successful use of a combination of nonsurgical root canal treatment and ultrasonic irrigation for collaborative management of a maxillary left lateral incisor with perforation of the apical third of the root. During the endodontic treatment procedure, the ultrasonically activated tip was used for intracanal irrigation. The area of perforation in the apical third of the root and the main root canal space were obturated with gutta-percha and root canal sealer, using a lateral condensation method. A follow-up clinical and radiographic examination at 5 years after treatment showed an asymptomatic tooth with excellent osseous healing.

  10. A tip-localized RhoGAP controls cell polarity by globally inhibiting Rho GTPase at the cell apex.

    PubMed

    Hwang, Jae-Ung; Vernoud, Vanessa; Szumlanski, Amy; Nielsen, Erik; Yang, Zhenbiao

    2008-12-23

    Highly elongated eukaryotic cells (e.g., neuronal axons, fungal hyphae, and pollen tubes) are generated through continuous apically restricted growth (tip growth), which universally requires tip-localized Rho GTPases. We used the oscillating pollen tube as a model system to determine the function and regulation of Rho GTPases in tip growth. Our previous work showed that the spatiotemporal dynamics of the apical cap of the activated Rho-like GTPase from Plant 1 (ROP1) are critical for tip growth in pollen tubes. However, the underlying mechanism for the generation and maintenance of this dynamic apical cap is poorly understood. A screen for mutations that enhance ROP1-overexpression-induced depolarization of pollen-tube growth identified REN1 (ROP1 enhancer 1) in Arabidopsis, whose null mutations turn elongated pollen tubes into bulbous cells. REN1 encodes a novel Rho GTPase-activating protein (RhoGAP) required for restricting the ROP1 activity to the pollen-tube tip. REN1 was localized to exocytic vesicles accumulated in the pollen-tube apex, as well as to the apical plasma membrane at the site of ROP1 activation. The apical localization of REN1 and its function in controlling growth polarity was compromised by disruption of ROP1-dependent F-actin and vesicular trafficking, which indicates that REN1 targeting and function is regulated by ROP1 downstream signaling. Our findings suggest that the REN1 RhoGAP controls a negative-feedback-based global inhibition of ROP1. This function provides a critical self-organizing mechanism, by which ROP signaling is spatially limited to the growth site and temporally oscillates during continuous tip growth. Similar spatiotemporal control of Rho GTPase signaling may also play an important role in cell-polarity control in other systems, including tip growth in fungi and cell movement in animals.

  11. Transport of salicylic acid through monolayers of a kidney epithelial cell line (LLC-PK1).

    PubMed

    Chatton, J Y; Roch-Ramel, F

    1992-05-01

    LLC-PK1 cells were used as a model of renal proximal epithelium to study the nonionic diffusion of salicylic acid (SAL). The apparent [14C]SAL transcellular permeability (PSal) and intracellular content were estimated at 20-21 degrees C from fluxes measured across cell monolayers grown on filters, in both apical-to-basolateral and basolateral-to-apical directions. The medium pH of the cis-side was varied from 6.0 to 7.4, and the medium pH of the trans-side was kept at 7.4. In the apical-to-basolateral direction, PSal increased linearly with the calculated concentration of nonionized SAL, indicating that SAL permeability was essentially the result of nonionic diffusion. In the basolateral-to-apical direction, PSal was about 2.5-fold higher than in the apical-to-basolateral direction and was not linearly related to the concentration of nonionized SAL molecules (0-4.5 nM), suggesting that besides nonionic diffusion, SAL was transported in its ionized form by a facilitated mechanism still active at 21 degrees C. This was confirmed by measuring basolateral-to-apical fluxes at 37 degrees C and observing that probenecid, an inhibitor of organic anion secretion, and cold SAL decreased PSal. Interestingly, at 37 degrees C, PSal in the apical-to-basolateral direction was also decreased by probenecid and cold SAL, suggesting the existence of a facilitated transport in this direction. These data demonstrated that the secretory transport of SAL is present in LLC-PK1 cells. The facilitated transport observed in the apical-to-basolateral direction suggests that in proximal tubule, SAL reabsorption might occur by facilitated mechanism and nonionic diffusion.

  12. [The effect of instrumentation on original apical foramen shape using steel vs niti rotatory: computerized analysis].

    PubMed

    Gallina, G; Cumbo, E; Gallo, P; Pizzo, G; D'Angelo, M

    2002-01-01

    A fundamental requirement to obtain a correct endodontic preparation depends on the respect, during cleaning and shaping, of the original morphology of the apical foramen, (AF), so that the filling material will form a complete seal. In our previous studies, in order to verify if this presumption was respected using rotary NiTi instruments with cutting tip, we instrumented extracted teeth characterised by a different degree of root curvatures. Using a standardized system to replace the sample, the original shape of the apical foramen of each tooth was recorded using a computerised technique and then compared to the shape after instrumentation. The data showed differences on AF shapes depending on the degree of root canal curvatures. In fact, the teeth with straight canals showed the least alterations on the original AF shape; on the contrary, the teeth with curved canals showed apical foramen enlargement or transportation. Therefore, in the current study we aimed to compare the in vitro effects of stainless steel, (Flexofile, Dentsply Maillefer, Baillaigues, Switzerland), vs NiTi safe cutting tip (Quantec SC, (Tycom Dental, Irvine CA, U:S.A.), vs NiTi non cutting tip, (Quantec LX, Tycom Dental, Irvine CA, U:S.A.), instrumentation on original apical foramen shape. We used NiTi instruments according to standard technique suggested by Tycom, and hand steel files, to instrument the apical third, according to the Crown-Down technique. Working length was fixed at -0.5 from AF. Our results suggested that in the presence of accentuated canal curves rotary Niti, with cutting tip, cause significantly more enlargement of the AF area. At the same time, we also observed that NiTi rotary files, with both cutting and non-cutting tip, cause eccentric enlargement of AF in curved canals. Therefore, NiTi engine-driven instruments should be used carefully in the presence of accentuated canal curves to avoid enlargement or transportation of AF, probably because rotary NiTi files may slip out of operative control, leaving a mark on the foramen shape. In the presence of severe curves, we suggest modifying the operative sequences by alternating rotary NiTi with hand NiTi or stainless steel instruments, especially in the preparation of apical third.

  13. Some Pecularities of the Graviresponse in Vaucheria

    NASA Technical Reports Server (NTRS)

    Gavrilova, O. V.; Rudanova, E. E.; Voloshko, L. N.; Gabova, A. V.

    1996-01-01

    The growth and position of nuclei in the siphonaceous alga Vaucheria sessilis were investigated under conditions of hypergravity and hypogravity. Under hypergravity conditions, active sporogenesis was observed. The accumulation of nuclei in the apical and branching zone preceeded the sporogenesis. The anti-microtubular agent, colchicine inhibits sporogenesis and the response of Vaucheria to hypergravity. Under hypogravity conditions, the quantity of nuclei increased throughout the whole branch. Colchicine prevents the migration of nuclei from the apical zone to the basal part of the branch. The anti-actin agent phalloidin prevents the formation of an actin network, and phalloidin-poisoned cells lose a cluster of nuclei in the apical zone. However, the gravity dependent response is less pronounced. It is supposed that, in Vaucheria sessilis, the primary stages of the reception and translation of gravitational signals coincide with those for light signals and active division of nuclei in the growth zone is an integral part of the graviresponse.

  14. A Comparative Study on the Amount of Extruded Material from the Apical Foramen with NiTi Rotary and Stainless Steel Hand Instruments

    PubMed Central

    Nazari, Shahrzad; MirMotalebi, Farshid

    2006-01-01

    INTRODUCTION: The purpose of the present study was to quantify and compare the amount of extruded debris from apical foramen after instrumentation of the root canal system with hand and rotary instruments. MATERIALS AND METHODS: Root canals of forty five fresh extracted single rooted human teeth with mature apexes and less than 15 degree of root curvature were instrumented in group A with stainless steel K-Type files, in group B with rotary NiTi Flex Master files, and in group C with rotary NiTi ProTaper files and followed weighting the extruded debris by a digital scale to within 0.0001 gram accuracy. RESULTS: In all groups, the mean weight of extruded debris was not more than 5 mg (P=0.0l) and was ranked as: Group A>Group B>Group C. There were statistically significant differences among three groups (p=0.0l). The mean value of extruded debris in the ProTaper and Flex Master groups were 0.652 and 0.788 mg, respectively. CONCLUSION: The study revealed that the amount of extruded debris from the apical foramen was minimal when ProTaper files were used. PMID:24470804

  15. Torque during canal instrumentation using rotary nickel-titanium files.

    PubMed

    Sattapan, B; Palamara, J E; Messer, H H

    2000-03-01

    Nickel-titanium engine-driven rotary instruments are used increasingly in endodontic practice. One frequently mentioned problem is fracture of an instrument in the root canal. Very few studies have been conducted on torsional characteristics of these instruments, and none has been done under dynamic conditions. The purposes of this study were to measure the torque generated and the apical force applied during instrumentation with a commercial engine-driven nickel-titanium file system, and to relate torque generated during simulated clinical use to torsional failure of the instruments. Ten extracted human teeth (five with small-sized and five with medium-sized straight root canals) were instrumented with Quantec Series 2000 files, and the torque and apical force generated were measured. The applied apical force was generally low, not exceeding 150 g in either small or medium canals. The torque depended on the tip size and taper of each instrument, and on canal size. Instruments with 0.05 and 0.06 taper generated the highest torque, which was greater in small than in medium canals. The torque at failure was significantly (p < 0.001) higher than torque during instrumentation, but with considerable variation in the extent of the difference.

  16. External apical root resorption in maxillary incisors in orthodontic patients: associated factors and radiographic evaluation

    PubMed Central

    Patanaporn, Virush; Janhom, Apirum; Korwanich, Narumanus

    2012-01-01

    Purpose This study was performed to evaluate the incidence and degree of external apical root resorption of maxillary incisors after orthodontic treatment and to evaluate particular associated factors related to external apical root resorption. Materials and Methods The records and maxillary incisor periapical radiographs of 181 patients were investigated. Crown and root lengths were measured and compared on the pre- and post-treatment periapical radiographs. Crown length was measured from the center of the incisal edge to the midpoint of the cemento-enamel junction (CEJ). Root length was measured from the CEJ midpoint to the root apex. A correction factor for the enlargement difference was used to calculate root resorption. Results The periapical radiographs of 564 teeth showed that the average root resorption was 1.39±1.27 (8.24±7.22%) and 1.69±1.14 mm (10.16±6.78%) for the maxillary central and lateral incisors, respectively. The results showed that the dilacerated or pointed roots, maxillary premolar extraction cases, and treatment duration were highly significant factors for root resorption (p<0.001). Allergic condition was a significant factor at p<0.01. Age at the start of treatment, large overjet, and history of facial trauma were also factors significantly associated with root resorption (p<0.05). There was no statistically significant difference in root resorption among the factors of gender, overbite, tongue-thrusting habit, types of malocclusion, and types of bracket. Conclusion These results suggested that orthodontic treatment should be carefully performed in pre-treatment extraction patients who have pointed or dilacerated roots and need long treatment duration. PMID:23071964

  17. Polyphenol-rich black chokeberry (Aronia melanocarpa) extract regulates the expression of genes critical for intestinal cholesterol flux in Caco-2 cells.

    PubMed

    Kim, Bohkyung; Park, Youngki; Wegner, Casey J; Bolling, Bradley W; Lee, Jiyoung

    2013-09-01

    Black chokeberry (Aronia melanocarpa) is a rich source of polyphenols. The hypolipidemic effects of polyphenol-rich black chokeberry extract (CBE) have been reported, but underlying mechanisms have not been well characterized. We investigated the effect of CBE on the expression of genes involved in intestinal lipid metabolism. Caco-2 cells were incubated with 50 or 100 μg/ml of CBE for 24 h for quantitative realtime polymerase chain reaction analysis. Expression of genes for cholesterol synthesis (3-hydroxy-3-methylglutaryl coenzyme A reductase and sterol regulatory element binding protein 2), apical cholesterol uptake (Niemann-Pick C1 Like 1 and scavenger receptor class B Type 1) and basolateral cholesterol efflux [ATP-binding cassette transporter A1 (ABCA1)] was significantly decreased by CBE compared with control. Western blot analysis confirmed that CBE inhibited expression of these proteins. In contrast, CBE markedly induced mRNA and/or protein levels of ABCG5 and ABCG8 that mediate apical cholesterol efflux to the intestinal lumen. Furthermore, CBE significantly increased mRNA and protein levels of low-density lipoprotein (LDL) receptor, and cellular LDL uptake. Expression of genes involved in lipid metabolism and lipoprotein assembly, including sterol regulatory element-binding protein 1c, fatty acid synthase and acyl-CoA oxidase 1, was significantly decreased by CBE in a dose-dependent manner. Concomitantly, CBE significantly increased sirtuin 1, 3 and 5 mRNA levels, while it decreased SIRT-2. Our data suggest that hypolipidemic effects of CBE may be attributed, at least in part, to increased apical efflux of LDL-derived cholesterol and to decreased chylomicron formation in the intestine; and specific isoforms of SIRT may play an important role in this process. Copyright © 2013 Elsevier Inc. All rights reserved.

  18. Control of apical membrane chloride permeability in the renal A6 cell line by nucleotides

    PubMed Central

    Banderali, U; Brochiero, E; Lindenthal, S; Raschi, C; Bogliolo, S; Ehrenfeld, J

    1999-01-01

    The effect of extracellular nucleotides applied on the apical side of polarised A6 cells grown on permeant filters was investigated by measuring the changes in (i) the 36Cl efflux through the apical membranes, (ii) the intracellular chloride concentrations (aCli, measured with N-(6-methoxyquinolyl) acetoethyl ester, MQAE), (iii) ICl, the short-circuit current in the absence of Na+ transport and (iv) the characteristics of the apical chloride channels using a patch-clamp approach. ATP or UTP (0.1-500 μm) transiently stimulated ICl. The sequence of purinergic agonist potencies was UTP = ATP > ADP >> the P2X-selective agonist β,γ-methylene ATP = the P2Y-selective agonist 2-methylthioATP. Suramin (100 μm) as the P2Y antagonist Reactive Blue 2 (10 μm) had no effect on the UTP (or ATP)-stimulated current. These findings are consistent with the presence of P2Y2-like receptors located on the apical membranes of A6 cells. Apical application of adenosine also transiently increased ICl. This effect was blocked by theophylline while the UTP-stimulated ICl was not. The existence of a second receptor, of the P1 type is proposed. ATP (or UTP)-stimulated ICl was blocked by apical application of 200 μmN-phenylanthranilic acid (DPC) or 100 μm niflumic acid while 100 μm glibenclamide was ineffective. Ionomycin and thapsigargin both transiently stimulated ICl; the nucleotide stimulation of ICl was not suppressed by pre-treatment with these agents. Chlorpromazin (50 μm), a Ca2+-calmodulin inhibitor strongly inhibited the stimulation of ICl induced either by apical UTP or by ionomycin application. BAPTA-AM pre-treatment of A6 cells blocked the UTP-stimulated ICl. Niflumic acid also blocked the ionomycin stimulated ICl. A fourfold increase in 36Cl effluxes through the apical membranes was observed after ATP or UTP application. These increases of the apical chloride permeability could also be observed when following aCli changes. Apical application of DPC (1 mm) or 5-nitro-2(3-phenylpropylamino)benzoic acid (NPPB; 500 μm) produced an incomplete inhibition of 36Cl effluxes through the apical membranes in ATP-stimulated and in untreated monolayers. In single channel patch-clamp experiments, an apical chloride channel with a unitary single channel conductance of 7.3 ± 0.6 pS (n = 12) was usually observed. ATP application induced the activation of one or more of these channels within a few minutes. These results indicate that multiple purinergic receptor subtypes are present in the apical membranes of A6 cells and that nucleotides can act as modulators of Cl− secretion in renal cells. PMID:10457087

  19. Osseointegration of dental implants in extraction sockets preserved with porous titanium granules - an experimental study.

    PubMed

    Verket, Anders; Lyngstadaas, Ståle P; Rønold, Hans J; Wohlfahrt, Johan C

    2014-02-01

    This study investigated osseointegration of dental implants inserted in healed extraction sockets preserved with porous titanium granules (PTG). Three adult female minipigs (Gøttingen minipig; Ellegaard A/S, Dalmose, Denmark) had the mandibular teeth P2, P3 and P4 extracted. The extraction sockets were preserved with metallic PTG (Tigran PTG; Tigran Technologies AB, Malmö, Sweden) n = 12, heat oxidized white porous titanium granules (WPTG) (Tigran PTG White) n = 12 or left empty (sham) n = 6. All sites were covered with collagen membranes (Bio-Gide; Geistlich Pharma, Wolhausen, Switzerland) and allowed 11 weeks of healing before implants (Straumann Bone Level; Straumann, Basel, Switzerland) were inserted. The temperature was measured during preparation of the osteotomies. Resonance frequency analysis (RFA, Osstell; Osstell AB, Gothenburg, Sweden) was performed at implant insertion and at termination. After 6 weeks of submerged implant healing, the pigs were euthanized and jaw segments were excised for microCT and histological analyses. In the temperature and RFA analyses no significant differences were recorded between the test groups. The microCT analysis demonstrated an average bone volume of 61.7% for the PTG group compared to 50.3% for the WPTG group (P = 0.03) and 57.1% for the sham group. Histomorphometry demonstrated an average bone-to-implant contact of 68.2% for the PTG group compared to 36.6% for the WPTG group and 60.9% for the sham group (n.s). Eight out of ten implants demonstrated apical osseous defects in the WPTG group, but similar defects were observed in all groups. PTG preserved extraction sockets demonstrate a similar outcome as the sham control group for all analyses suggesting that this material potentially can be used for extraction socket preservation prior to implant installment. Apical osseous defects were however observed in all groups including the sham group, and a single cause could not be determined. © 2012 John Wiley & Sons A/S.

  20. Caspase enzymology and activation mechanisms.

    PubMed

    Mace, Peter D; Riedl, Stefan J; Salvesen, Guy S

    2014-01-01

    Apical caspases 8, 9, and 10 are only active as dimers. These dimers are unstable, and to characterize their activity they need to be maintained in vitro in a dimeric state. We provide updated methods for those looking to characterize various aspects of caspase function. We describe full methods for those looking to activate caspases in vitro using kosmotropic reagents, an essential step in characterizing upstream (apical) caspases. We detail methods for fusion of caspase domains to engineered dimerization domains as an alternative method to trigger regulated dimerization of caspases. We also describe methods to determine caspase activity profiles in cells and provide methods for studying the ability of SMAC-mimetic reagents to release inhibition of caspases by IAPs. © 2014 Elsevier Inc. All rights reserved.

  1. A method to generate the surface cell layer of the 3D virtual shoot apex from apical initials.

    PubMed

    Kucypera, Krzysztof; Lipowczan, Marcin; Piekarska-Stachowiak, Anna; Nakielski, Jerzy

    2017-01-01

    The development of cell pattern in the surface cell layer of the shoot apex can be investigated in vivo by use of a time-lapse confocal images, showing naked meristem in 3D in successive times. However, how this layer is originated from apical initials and develops as a result of growth and divisions of their descendants, remains unknown. This is an open area for computer modelling. A method to generate the surface cell layer is presented on the example of the 3D paraboloidal shoot apical dome. In the used model the layer originates from three apical initials that meet at the dome summit and develops through growth and cell divisions under the isotropic surface growth, defined by the growth tensor. The cells, which are described by polyhedrons, divide anticlinally with the smallest division plane that passes depending on the used mode through the cell center, or the point found randomly near this center. The formation of the surface cell pattern is described with the attention being paid to activity of the apical initials and fates of their descendants. The computer generated surface layer that included about 350 cells required about 1200 divisions of the apical initials and their derivatives. The derivatives were arranged into three more or less equal clonal sectors composed of cellular clones at different age. Each apical initial renewed itself 7-8 times to produce the sector. In the shape and location and the cellular clones the following divisions of the initial were manifested. The application of the random factor resulted in more realistic cell pattern in comparison to the pure mode. The cell divisions were analyzed statistically on the top view. When all of the division walls were considered, their angular distribution was uniform, whereas in the distribution that was limited to apical initials only, some preferences related to their arrangement at the dome summit were observed. The realistic surface cell pattern was obtained. The present method is a useful tool to generate surface cell layer, study activity of initial cells and their derivatives, and how cell expansion and division are coordinated during growth. We expect its further application to clarify the question of a number and permanence or impermanence of initial cells, and possible relationship between their shape and oriented divisions, both on the ground of the growth tensor approach.

  2. Dopamine Induces LTP Differentially in Apical and Basal Dendrites through BDNF and Voltage-Dependent Calcium Channels

    ERIC Educational Resources Information Center

    Navakkode, Sheeja; Sajikumar, Sreedharan; Korte, Martin; Soong, Tuck Wah

    2012-01-01

    The dopaminergic modulation of long-term potentiation (LTP) has been studied well, but the mechanism by which dopamine induces LTP (DA-LTP) in CA1 pyramidal neurons is unknown. Here, we report that DA-LTP in basal dendrites is dependent while in apical dendrites it is independent of activation of L-type voltage-gated calcium channels (VDCC).…

  3. Phytochrome mediated gravimorphogenesis in the moss protonemata

    NASA Astrophysics Data System (ADS)

    Demkiv, O.; Khorkavtsiv, Y.

    Moss protonemata are among the few cell types known that both sense and respond to gravity and light. Apical cells of moss protonemata grow by oriented tip growth which is negatively gravitropic in the dark or positively phototropic in unilateral light. Gametophore buds always arise some 4 - 6 cells behind the apical cells of main protonemal filaments differentiating from apical cells of shortened side branches. It has been shown, however, that in Pohlia nutans, as in Pottia intermedia (Ripetskyj et al, 1997) the apical cells of main filaments of dark grown protonemata differentiate buds under the influence of light. We tested the effectiveness of white and monochromatic light of the visible spectrum on the bud formation of Pohlia nutans which had been grown in darkness. The most morphogenetically effective light was red light, but green, yellow and white light were also active. Blue light alone completely inhibits bud formation but supplemented with red light this inhibitory effect of blue light is couneracted and buds are formed, provided a minimum exposure of red light is maintained. M.Bopp (1985) had demosnrated that exclusively cell that had not reached 80 μm in length initiated bud formation. Red light seemed to inhibit growth of apical cells and to promote bud formation. The action spectra for the induction of buds formation are, as expected, very similar to the absorption spectra of Pf r . The relatively small effectivity of quanta in the short-wave spectral range is caused by the strong absorption of radiation of < 520 nm by carotenoids and flavines. Red light might act as a triger for morphogenetic processes in dark-grown cells. The white or red light stopped an elongation of main filaments apical cells of protonema grown in the dark for 7 days retaining the rate of the cell divisions practically constant. As a result short apical cells are formed ready for a transition to new morphogenetic pathway of gametophore buds formation. The reaction proved to be reversible one. Transfering of the moss protonemata after 2 days illumination back into the dark leads to dedifferentiation of the apical bud cells into apical protonemal cells. Clinostat rotation of the dark-grown protonemata partially ihibited the light- dependent transformation of their apical cells into gameophore buds. The mechanisms of transformation of apical into the gametophore buds in the Pohlia nutans protonemata are discussed.

  4. Water Permeability Adjusts Resorption in Lung Epithelia to Increased Apical Surface Liquid Volumes.

    PubMed

    Schmidt, Hanna; Michel, Christiane; Braubach, Peter; Fauler, Michael; Neubauer, Daniel; Thompson, Kristin E; Frick, Manfred; Mizaikoff, Boris; Dietl, Paul; Wittekindt, Oliver H

    2017-03-01

    The apical surface liquid (ASL) layer covers the airways and forms a first line of defense against pathogens. Maintenance of ASL volume by airway epithelia is essential for maintaining lung function. The proteolytic activation of epithelial Na + channels is believed to be the dominating mechanism to cope with increases in ASL volumes. Alternative mechanisms, in particular increases in epithelial osmotic water permeability (P osm ), have so far been regarded as rather less important. However, most studies mainly addressed immediate effects upon apical volume expansion (AVE) and increases in ASL. This study addresses the response of lung epithelia to long-term AVE. NCI-H441 cells and primary human tracheal epithelial cells, both cultivated in air-liquid interface conditions, were used as models for the lung epithelium. AVE was established by adding isotonic solution to the apical surface of differentiated lung epithelia, and time course of ASL volume restoration was assessed by the deuterium oxide dilution method. Concomitant ion transport was investigated in Ussing chambers. We identified a low resorptive state immediately after AVE, which coincided with proteolytic ion transport activation within 10-15 minutes after AVE. The main clearance of excess ASL occurred during a delayed (hours after AVE) high resorptive state, which did not correlate with ion transport activation. Instead, high resorptive state onset coincided with an increase in P osm , which depended on aquaporin up-regulation. In summary, our data demonstrate that, aside from ion transport activation, modulation of P osm is a major mechanism to compensate for long-term AVE in lung epithelia.

  5. False Broomrape: A Physiological Disorder Caused by Growth-Regulator Imbalance 1

    PubMed Central

    Hamilton, James L.; Lowe, Richie H.; Skoog, Folke

    1972-01-01

    False broomrape on tobacco (Nicotiana tabacum) can be induced by applying cytokinins to the roots, by causing an increase in the cytokinin-auxin ratio in the roots, by removing the apical and auxillary buds, or by applying extracts from tobacco with false broomrape to the roots of healthy plants. It can be prevented by treating debudded plants with auxin. Images PMID:16658161

  6. Deproteinized Bovine Bone Mineral or Autologous Bone at Dehiscence Type Defects at Implants Installed Immediately into Extraction Sockets: An Experimental Study in Dogs.

    PubMed

    Pereira, Flavia Priscila; De Santis, Enzo; Hochuli-Vieira, Eduardo; de Souza Faco, Eduardo F; Pantani, Fabio; Salata, Luiz A; Botticelli, Daniele

    2016-06-01

    The aim of this study was to evaluate bone regeneration at surgically created dehiscence buccal defects at implants placed immediately into extraction sockets (IPIES) of small dimensions filled with autogenous bone or deproteinized bovine bone mineral (DBBM) associated with a collagen membrane. Eight Labrador dogs were used and implants were placed immediately into the extraction sockets of the second premolar. The buccal wall was subsequently removed to create a standardized defect, 4 mm wide coronally, 2 mm wide apically, and 6 mm high. Autogenous bone particles (AB) or DBBM granules were used to fill the defects. All surgical sites were subsequently covered by a resorbable collagen membrane and a non-submerged healing was allowed. After 4 months, the animals were euthanized and bone blocks harvested and processed for histomorphometric analysis. The bony crest at the buccal aspect (C) was located 2.3 ± 0.8 mm and 1.7 ± 0.7 mm apically to the implant shoulder (IS) at the AB and DBBM sites, respectively. The coronal levels of osseointegration at the buccal aspect (B) were located 2.7 ± 0.7 mm and 2.2 ± 1.0 mm apically to IS at the AB and DBBM sites, respectively. At the AB sites, the peri-implant mucosa was located 4.3 ± 0.9 mm, 4.7 ± 0.9 mm, and 2.0 ± 1.6 mm coronally to C, B, and IS, respectively. The corresponding values at the DBBM sites were 4.3 ± 0.6 mm, 4.8 ± 0.6 mm, and 2.5 ± 0.8 mm, respectively. No statistically significant differences were found. The treatment of surgically created buccal defects at IPIES sites using Bio-Oss® (Geistlich Biomaterials, Wolhusen, LU, Switzerland) or autogenous bone, concomitantly with a collagen membrane, engenders bone regeneration to a similar extent after 4 months of healing. © 2015 Wiley Periodicals, Inc.

  7. Automated extraction of single H atoms with STM: tip state dependency

    NASA Astrophysics Data System (ADS)

    Møller, Morten; Jarvis, Samuel P.; Guérinet, Laurent; Sharp, Peter; Woolley, Richard; Rahe, Philipp; Moriarty, Philip

    2017-02-01

    The atomistic structure of the tip apex plays a crucial role in performing reliable atomic-scale surface and adsorbate manipulation using scanning probe techniques. We have developed an automated extraction routine for controlled removal of single hydrogen atoms from the H:Si(100) surface. The set of atomic extraction protocols detect a variety of desorption events during scanning tunneling microscope (STM)-induced modification of the hydrogen-passivated surface. The influence of the tip state on the probability for hydrogen removal was examined by comparing the desorption efficiency for various classifications of STM topographs (rows, dimers, atoms, etc). We find that dimer-row-resolving tip apices extract hydrogen atoms most readily and reliably (and with least spurious desorption), while tip states which provide atomic resolution counter-intuitively have a lower probability for single H atom removal.

  8. A local autocrine axis in the testes that regulates spermatogenesis

    PubMed Central

    Cheng, C. Yan; Mruk, Dolores D.

    2014-01-01

    Spermiation—the release of mature spermatozoa from Sertoli cells into the seminiferous tubule lumen—occurs by the disruption of an anchoring device known as the apical ectoplasmic specialization (apical ES). At the same time, the blood–testis barrier (BTB) undergoes extensive restructuring to facilitate the transit of preleptotene spermatocytes. While these two cellular events take place at opposite ends of the Sertoli cell epithelium, the events are in fact tightly coordinated, as any disruption in either process will lead to infertility. A local regulatory axis exists between the apical ES and the BTB in which biologically active laminin fragments produced at the apical ES by the action of matrix metalloproteinase 2 can regulate BTB restructuring directly or indirectly via the hemidesmosome. Equally important, polarity proteins play a crucial part in coordinating cellular events within this apical ES–BTB–hemidesmosome axis. Additionally, testosterone and cytokines work in concert to facilitate BTB restructuring, which enables the transit of spermatocytes while maintaining immunological barrier function. Herein, we will discuss this important autocrine-based cellular axis that parallels the hormonal-based hypothalamic–pituitary–testicular axis that regulates spermatogenesis. This local regulatory axis is the emerging target for male contraception. PMID:20571538

  9. Effects of Osmolality on Paracellular Transport in MDCK II Cells

    PubMed Central

    Hirai, Toyohiro; Furuse, Mikio

    2016-01-01

    Epithelia separate apical and basal compartments, and movement of substances via the paracellular pathway is regulated by tight junctions. Claudins are major constituents of tight junctions and involved in the regulation of tight junction permeability. On the other hand, the osmolality in the extracellular environment fluctuates in association with life activity. However, effects of osmotic changes on the permeaibility of claudins are poorly understood. Therefore, we investigated the effects of osmotic changes on the paracellular transport in MDCK II cells. Interestingly, apical hyposmolality decreased cation selectivity in the paracellular pathway gradually with time, and the elimination of the osmotic gradient promptly restored the cation selectivity. Apical hyposmolality also induced bleb formation at cell-cell contacts and changed the shape of cell-cell contacts from a jagged pattern to a slightly linear pattern. In claudin-2 knockout MDCK II cells, the decrease of cation selectivity, the bleb formation, nor the changes in the shape of cell-cell contacts was observed under the apical hyposmolality. Our findings in this study indicate that osmotic gradient between apical and basal sides is involved in the acute regulation of the cation selective property of claudin-2 channels. PMID:27855213

  10. Ion and lipid signaling in apical growth-a dynamic machinery responding to extracellular cues.

    PubMed

    Malhó, Rui; Serrazina, Susana; Saavedra, Laura; Dias, Fernando V; Ul-Rehman, Reiaz

    2015-01-01

    Apical cell growth seems to have independently evolved throughout the major lineages of life. To a certain extent, so does our body of knowledge on the mechanisms regulating this morphogenetic process. Studies on pollen tubes, root hairs, rhizoids, fungal hyphae, even nerve cells, have highlighted tissue and cell specificities but also common regulatory characteristics (e.g., ions, proteins, phospholipids) that our focused research sometimes failed to grasp. The working hypothesis to test how apical cell growth is established and maintained have thus been shaped by the model organism under study and the type of methods used to study them. The current picture is one of a dynamic and adaptative process, based on a spatial segregation of components that network to achieve growth and respond to environmental (extracellular) cues. Here, we explore some examples of our live imaging research, namely on cyclic nucleotide gated ion channels, lipid kinases and syntaxins involved in exocytosis. We discuss how their spatial distribution, activity and concentration suggest that the players regulating apical cell growth may display more mobility than previously thought. Furthermore, we speculate on the implications of such perspective in our understanding of the mechanisms regulating apical cell growth and their responses to extracellular cues.

  11. Apical and basal membrane ion transport mechanisms in bovine retinal pigment epithelium.

    PubMed

    Joseph, D P; Miller, S S

    1991-04-01

    1. Intracellular voltage recordings using conventional and double-barrelled chloride-selective microelectrodes have been used to identify several transport mechanisms at the apical and basolateral membranes of the isolated bovine retinal pigment epithelium (RPE)-choroid preparation. Intracellular recordings were obtained from two cell populations, melanotic (pigmented) and amelanotic (non-pigmented). The electrical properties of these two populations are practically identical. For melanotic cells the average apical resting membrane potential (VA) is -61 +/- 2 mV (mean +/- S.E.M., n = 49 cells, thirty-three eyes). For these cells the ratio of apical to basolateral membrane resistance (a) was 0.22 +/- 0.02. The mean transepithelial voltage and resistance were 6 +/- 1 mV and 138 +/- 7 omega cm2, respectively. 2. The apical membrane, which faces the distal retina, contains a Ba(2+)-inhibitable K+ conductance and a ouabain-inhibitable, electrogenic Na(+)-K+ pump. In addition it contains a bumetanide-sensitive mechanism, the putative Na(+)-K(+)-Cl- cotransporter. The basolateral membrane contains a DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonic acid)-inhibitable chloride channel. The relative conductances of the apical and basolateral membranes to K+ and Cl- are TK approximately 0.9 and TCl approximately 0.7, respectively. 3. The ouabain-induced fast phase of apical membrane depolarization (0-30 s) was used to calculate the equivalent resistances of the apical (RA) and basolateral (RB) cell membranes, as well as the paracellular or shunt resistance (RS). They are: 3190 +/- 400, 17920 +/- 2730 and 2550 +/- 200 omega (mean +/- S.E.M., n = 9 tissues), respectively. From these data the equivalent electromotive forces (EMF) at the apical (EA) and basolateral (EB) membranes were also calculated. They are: -69 +/- 5.0 and -24 +/- 5.0 mV, respectively. 4. Intracellular Cl- activity (aiCl) was measured using double-barreled ion-selective microelectrodes. In the steady state aiCl = 61 +/- 4.0 mM and the Nernst potential ECl = -13.5 +/- 1.5 mV (mean +/- S.E.M., n = 4). 5. In the intact eye or in retina, RPE-choroid preparations it has been shown that the transition between light and dark alters the K+ concentration in the extracellular (or subretinal) space between the photoreceptors and the apical membrane of the RPE. These light-induced changes in subretinal [K+]o were qualitatively simulated in vitro by altering apical K+ between 5 and 2 mM. This produced a sequence of voltage changes at the apical and basolateral membranes that had three operationally distinct phases. Phase 1 is generated by the combination of an apical membrane K+ diffusion potential and inhibition of the electrogenic Na(+)-K+ pump.(ABSTRACT TRUNCATED AT 400 WORDS)

  12. Characterization of active ion transport across primary rabbit corneal epithelial cell layers (RCrECL) cultured at an air-interface.

    PubMed

    Chang-Lin, Joan-En; Kim, Kwang-Jin; Lee, Vincent H L

    2005-06-01

    Previously, we reported the development of a primary culture model of tight rabbit corneal epithelial cell layers (RCrECL) characterizing bioelectric parameters, morphology, cytokeratin, and passive permeability. In the present study, we specifically evaluated the active ion transport processes of RCrECL cultured from either pigmented or albino rabbits. Primary cultured RCrECL were grown at an air-interface on Clear-Snapwells precoated with collagen/fibronectin/laminin and mounted in a modified Ussing-type chamber for the evaluation of their active ion transport processes under short-circuited conditions. Contribution of active Na(+) and Cl(-) transport to overall short-circuit current (I(sc)) was evaluated by removing Na(+) and Cl(-), respectively, from bathing fluids of RCrECL and measurements of net fluxes of Na(+) and Cl(-) using (22)Na and (36)Cl, respectively. Amiloride and benzamil were used to determine the role of apical Na(+)-channel activities to net Na(+) fluxes. N-phenylanthranilic acid (NPAA), ouabain, BaCl(2) and bumetanide were used to determine the role of basolateral Na,K-ATPase, apical Cl(-)-channel, and basolateral K(+)-channel and Na(+)(K(+))2Cl(-)-cotransporter activities, respectively, in active ion transport across RCrECL. I(sc) of RCrECL derived from pigmented rabbits was comprised of 64+/-2% and 44+/-5% for active Na(+) and Cl(-) transport, respectively, consistent with net Na(+) absorption and Cl(-) secretion of 0.062+/-0.006 and 0.046+/-0.008 muEq/cm(2)/hr estimated from radionuclide fluxes. Apical amiloride and benzamil inhibited I(sc) by up to approximately 50% with an IC(50) of 1 and 0.1 microm, respectively, consistent with participation of apical epithelial Na(+)-channels to net Na(+) absorption across RCrECL cultured from pigmented rabbits. Addition of ouabain to the basolateral, NPAA to the apical, BaCl(2) to the basolateral and bumetanide to basolateral fluid decreased I(sc) by 86+/-1.5%, 53+/-3%, 18+/-1.8% and 13+/-1.9% in RCrECL cultured from pigmented rabbits, while 85+/-0.7%, 36+/-1.6%, 38+/-1.8% and 15+/-3.5% decreases are observed for RCrECL from albino rabbits, respectively. Air-interface cultured RCrECL from either pigmented or albino rabbits exhibited active ion transport properties similar to those present in excised tissues. This primary culture system may be a reliable in-vitro model for mechanistic characterization of corneal epithelial function and regulation of transport properties.

  13. Caspase selective reagents for diagnosing apoptotic mechanisms.

    PubMed

    Poreba, Marcin; Groborz, Katarzyna; Navarro, Mario; Snipas, Scott J; Drag, Marcin; Salvesen, Guy S

    2018-05-10

    Apical caspases initiate and effector caspases execute apoptosis. Reagents that can distinguish between caspases, particularly apical caspases-8, 9, and 10 are scarce and generally nonspecific. Based upon a previously described large-scale screen of peptide-based caspase substrates termed HyCoSuL, we sought to develop reagents to distinguish between apical caspases in order to reveal their function in apoptotic cell death paradigms. To this end, we selected tetrapeptide-based sequences that deliver optimal substrate selectivity and converted them to inhibitors equipped with a detectable tag (activity-based probes-ABPs). We demonstrate a strong relationship between substrate kinetics and ABP kinetics. To evaluate the utility of selective substrates and ABPs, we examined distinct apoptosis pathways in Jurkat T lymphocyte and MDA-MB-231 breast cancer lines triggered to undergo cell death via extrinsic or intrinsic apoptosis. We report the first highly selective substrate appropriate for quantitation of caspase-8 activity during apoptosis. Converting substrates to ABPs promoted loss-of-activity and selectivity, thus we could not define a single ABP capable of detecting individual apical caspases in complex mixtures. To overcome this, we developed a panel strategy utilizing several caspase-selective ABPs to interrogate apoptosis, revealing the first chemistry-based approach to uncover the participation of caspase-8, but not caspase-9 or -10 in TRAIL-induced extrinsic apoptosis. We propose that using select panels of ABPs can provide information regarding caspase-8 apoptotic signaling more faithfully than can single, generally nonspecific reagents.

  14. Apices of maxillary premolars observed by swept source optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Ebihara, Arata; Iino, Yoshiko; Yoshioka, Toshihiko; Hanada, Takahiro; Sunakawa, Mitsuhiro; Sumi, Yasunori; Suda, Hideaki

    2015-02-01

    Apicoectomy is performed for the management of apical periodontitis when orthograde root canal treatment is not possible or is ineffective. Prior to the surgery, cone beam computed tomography (CBCT) examination is often performed to evaluate the lesion and the adjacent tissues. During the surgical procedure, the root apex is resected and the resected surface is usually observed under dental operating microscope (DOM). However, it is difficult to evaluate the details and the subsurface structure of the root using CBCT and DOM. A new diagnostic system, swept source optical coherence tomography (SS-OCT), has been developed to observe the subsurface anatomical structure. The aim of this study was to observe resected apical root canals of human maxillary premolars using SS-OCT and compare the findings with those observed using CBCT and DOM. Six extracted human maxillary premolars were used. After microfocus computed tomography (Micro CT; for gold standard) and CBCT scanning of the root, 1 mm of the apex was cut perpendicular to the long axis of the tooth. Each resected surface was treated with EDTA, irrigated with saline solution, and stained with methylene blue dye. The resected surface was observed with DOM and SS-OCT. This sequence was repeated three times. The number of root canals was counted and statistically evaluated. There was no significant difference in the accuracy of detecting root canals among CBCT, DOM and SS-OCT (p > 0.05, Wilcoxon test). Because SS-OCT can be used in real time during surgery, it would be a useful tool for observing resected apical root canals.

  15. Transport characteristics of L-citrulline in renal apical membrane of proximal tubular cells.

    PubMed

    Mitsuoka, Keisuke; Shirasaka, Yoshiyuki; Fukushi, Akimasa; Sato, Masanobu; Nakamura, Toshimichi; Nakanishi, Takeo; Tamai, Ikumi

    2009-04-01

    L-Citrulline has diagnostic potential for renal function, because its plasma concentration increases with the progression of renal failure. Although L-citrulline extracted by glomerular filtration in kidney is mostly reabsorbed, the mechanism involved is not clearly understood. The present study was designed to characterize L-citrulline transport across the apical membranes of renal epithelial tubular cells, using primary-cultured rat renal proximal tubular cells, as well as the human kidney proximal tubular cell line HK-2. L-Citrulline was transported in a Na(+)-dependent manner from the apical side of both cell types cultured on permeable supports with a microporous membrane. Kinetic analysis indicated that the transport involves two distinct Na(+)-dependent saturable systems and one Na(+)-independent saturable system in HK-2 cells. The uptake was competitively inhibited by neutral and cationic, but not anionic amino acids. Relatively large cationic and anionic compounds inhibited the uptake, but smaller ones did not. In HK-2 cells, mRNA expression of SLC6A19 and SLC7A9, which encode B(0)AT1 and b(0,+)AT, respectively, was detected by RT-PCR. In addition, L-citrulline transport was significantly decreased in HK-2 cells in which either SLC6A19 or SLC7A9 was silenced. Hence, these results suggest that amino acid transporters B(0)AT1 and b(0,+)AT are involved in the reabsorption of L-citrulline in the kidney, at least in part, by mediating the apical membrane transport of L-citrulline in renal tubule cells.

  16. Apical Extrusion of Intracanal Bacteria following use of Two Engine-driven Instrumentation Techniques: An in vitro Study.

    PubMed

    Mohammed, Niyaz; Noushad, M C; Balan, Biji; Dhanesh, N; Jayasheelan, Nishi; Revankar, Vanita D

    2016-11-01

    The aim of the present study was to compare in vitro the amount of debris extruded apically from extracted teeth, using Twisted files and ProTaper rotary during two different instrumentation systems. Forty-five human single-rooted mandibular premolar teeth were randomly divided into three groups and contaminated with Enterococcus faecalis. The teeth in experimental groups were instrumented until reaching the working length with ProTaper rotary instruments and Twisted files with XSmart and XSmart Dual groups. Debris extruded from the apical foramen was collected into glass vials and the amount of bacteria was calculated. The data obtained were analyzed using Kruskal-Wallis one-way analysis of variance and Mann-Whitney U tests, with p = 0.05 as the level for statistical significance. The XSmart Dual group extruded comparatively lesser bacteria compared to the XSmart group. Lesser amount of bacterial extrusion was seen when Twisted files were used compared to the ProTaper files (p < 0.05). Under the circumstances of this in vitro study, it can be concluded that all instrumentation techniques produced measurable apical extrusion of debris. So, it is upon the practitioner to decide which system best fits their individual needs and their level of skill and experience that will provide the best possible endodontic care for our patients. The newer instrument designs, including noncutting tips, different cross sections, radial lands, and variable tapers, are better for the clinicians to improve working safety, to reduce the working time, and to create a greater flare within the preparations.

  17. External apical root resorption diagnosis by using FII human dentine fraction and salivary IGg.

    PubMed

    Da-Costa, Tânia Maris Pedrini Soares; Hidalgo, Mirian Marubayashi; Consolaro, Alberto; Lima, Carlos Eduardo de Oliveira; Tanaka, Evelise Ono; Itano, Eiko Nakagawa

    2018-06-01

    External apical root resorption as a consequence of orthodontic treatment is an inflammatory pathological process that results in permanent loss of tooth structure from the root apex. This study aimed to investigate the diagnostic potential of human dentine fractions and salivary IgG in external apical root resorption. Saliva samples were collected from 10 patients before (T0) and after 3 (T3), 6 (T6) and 12 (T12) months of orthodontic treatment. The total dentinal extract, obtained from human third molars, was fractioned by gel filtration chromatography in three fractions denominated FI, FII and FIII. The root resorption analysis of the upper central incisors was performed by digital image subtraction method. Reactivity of salivary IgG to antigenic fractions of dentine was determined by enzyme-linked immunosorbent assay (Elisa). Regardless of treatment, FI dentin fraction with high MM (<300kDa) was the one that presented highest reactivity with salivary IgG. However, it was found higher salivary IgG reactivity for FII (69 to 45 kilodalton [kDa]) as compared to FIII (<45kDa) at (T6) and (T12), (P<0.05), the same periods in that the root resorptions were detected. Our results suggest that FII human dentine fraction and salivary IgG have potential to be used in diagnosis and monitoring of external apical root resorption. The development of a practical and accessible biochemical test using saliva and FII dentine fraction may help in the prevention of severe root resorption. Copyright © 2018. Published by Elsevier Masson SAS.

  18. Film and digital periapical radiographs for the measurement of apical root shortening.

    PubMed

    El-Angbawi, Ahmed M F; McIntyre, Grant T; Bearn, David R; Thomson, Donald J

    2012-12-01

    The aim of this study was to compare the accuracy and agreement of scanned film and digital periapical radiographs for the measurement of apical root shortening. Twenty-four film and digital [phosphor plate sensor (PPS)] periapical radiographs were taken using the long-cone paralleling technique for six extracted teeth before and after 1mm of apical root trimming. All teeth were mounted using a typodont and the radiographs were recorded using a film holder and polysiloxane occlusal index for each tooth to ensure standardization during the different radiographic exposures. The film radiographs were scanned and the tooth length measurements for the scanned film and digital (PPS) images were calculated using Image-J-Link 1.4 software (http://rebweb.nih.gov/ij/index.html) for the two groups. The accuracy and agreement among the tooth length measurements from each group and the true tooth length measurements were calculated using intra-class correlation (ICC) tests and Bland and Altman plots. A high level of agreement was found between the true tooth length measurements and the scanned film measurements (ICC=0.979, limit of agreement 0.579 to -0.565) and the digital (PPS) radiograph measurements (ICC= 0.979, limit of agreement 0.596 to -0.763). Moreover, a high level of agreement was found between the scanned film and digital (PPS) radiographs for the measurement of tooth length ICC=0.991, limit of agreement 0.411-0.231. Film and digital (PPS) periapical radiographs are accurate methods for measuring apical root shortening with a high level of agreement. Key words:Root shortening, measurement, periapical radiographs, film, digital.

  19. Evaluation of debris extruded apically during the removal of root canal filling material using ProTaper, D-RaCe, and R-Endo rotary nickel-titanium retreatment instruments and hand files.

    PubMed

    Topçuoğlu, Hüseyin Sinan; Aktı, Ahmet; Tuncay, Öznur; Dinçer, Asiye Nur; Düzgün, Salih; Topçuoğlu, Gamze

    2014-12-01

    The aim of this study was to evaluate the amount of debris extruded apically during the removal of root canal filling material using ProTaper (Dentsply Maillefer, Ballaigues, Switzerland), D-RaCe (FKG Dentaire, La Chaux-de-Fonds, Switzerland), and R-Endo (Micro-Mega, Besançon, France) nickel-titanium (NiTi) rotary retreatment instruments and hand files. Sixty extracted single-rooted mandibular premolar teeth were prepared with K-files and filled with gutta-percha and AH Plus sealer (Dentsply DeTrey, Konstanz, Germany). The teeth were then randomly assigned to 4 groups (n = 15 for each group) for retreatment. The removal of canal filling material was performed as follows: hand files, ProTaper, D-RaCe, and R-Endo retreatment instruments. Debris extruded apically during the removal of canal filling material was collected into preweighed Eppendorf tubes. The tubes were then stored in an incubator at 70°C for 5 days. The weight of the dry extruded debris was established by subtracting the preretreatment and postretreatment weight of the Eppendorf tubes for each group. The data obtained were analyzed using 1-way analysis of variance and Tukey post hoc tests. All retreatment techniques caused the apical extrusion of debris. Hand files produced significantly more debris when compared with ProTaper, D-RaCe, and R-Endo rotary systems (P < .05). There was no statistical difference between the ProTaper, D-RaCe, and R-Endo retreatment systems (P > .05). The findings showed that during the removal of root canal filling material, rotary NiTi retreatment instruments used in this study caused less apical extrusion of debris compared with hand files. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  20. Effects of sodium hypochlorite associated with EDTA and etidronate on apical root transportation.

    PubMed

    Silva e Souza, P A R; das Dores, R S E; Tartari, T; Pinheiro, T P S; Tuji, F M; Silva e Souza, M H

    2014-01-01

    To evaluate the influence of sodium hypochlorite associated with EDTA and etidronate on apical root transportation. Forty-five roots of human mandibular molars with curvatures of 15-25° were embedded in acrylic resin to allow standardized angulation of the initial and final radiographs. The pre-instrumentation radiographs of the mesiobuccal canal of each root were taken using a radiograph digital sensor with a size 15 K-file in the canal. The canals were prepared with the ProTaper Universal system (Dentsply Maillefer, Ballaigues, Switzerland), using one of the following irrigation regimens during the instrumentation (n = 15): G1 - irrigation with 20 mL of saline solution (control); G2 - alternating irrigation with 2.5% hypochlorite solution (NaOCl) (15 mL); and 17% ethylenediaminetetraacetic acid (EDTA) (5 mL). During instrumentation, the canal was filled with NaOCl and then between each exchange of instrument filled with EDTA for 1 min, and G3 - irrigation with 20 mL of 5% NaOCl and 18% etidronate solution (HEBP) mixed in equal parts. The postinstrumentation radiographs were made with a F3 instrument in the canal. The images were magnified and superposed with Adobe Photoshop software (Adobe Systems, Mountain View, CA, USA). Apical transportation was determined with AutoCAD 2012 software (Autodesk Inc., San Rafael, CA, USA) by measuring the distance in millimetres between the tips of the instruments. The results were subjected to the nonparametric statistical Kruskal-Wallis test (α < 0.05). The median transportation and interquartile range values were 0.00 ± 0.05 for G1, 0.08 ± 0.23 for G2 and 0.13 ± 0.14 for G3. Comparison between groups showed that apical transportation in G3 was significantly greater than in G1 (P < 0.05). The use of NaOCl associated with etidronate increased apical transportation in the canals of extracted teeth. © 2013 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  1. Influence of a glide path on apical extrusion of debris during canal preparation using single-file systems in curved canals.

    PubMed

    Topçuoğlu, H S; Düzgün, S; Akpek, F; Topçuoğlu, G; Aktı, A

    2016-06-01

    To evaluate the effect of a glide path on the amount of apically extruded debris during canal preparation using single-file systems in curved canals. Ninety extracted mandibular molar teeth were randomly assigned to six groups (n = 15 for each group) for canal instrumentation. Endodontic access cavities were prepared in each tooth. In three of the six groups, a glide path was not created whereas a glide path was created using PathFile instruments on the mesial canals of all teeth in the remaining three groups. The mesial canals of the teeth were then instrumented with the following single-file instrument systems: WaveOne, Reciproc and OneShape. Debris extruded apically during instrumentation was collected into pre-weighed Eppendorf tubes. The tubes were then stored in an incubator at 70 °C for 5 days. The weight of the dry extruded debris was established by subtracting the pre-instrumentation and post-instrumentation weight of the Eppendorf tubes for each group. The data obtained were analysed using one-way analysis of variance (anova) and Tukey's post hoc tests. The OneShape file was associated with less debris extrusion than the Reciproc and WaveOne files when canal instrumentation was performed without a glide path (P < 0.05). However, no significant difference was found between the Reciproc and WaveOne files (P > 0.05). There was no significant difference amongst the OneShape, Reciproc and WaveOne files when a glide path was created before canal preparation in curved root canals (P > 0.05). All systems extruded significantly less debris in groups with a glide path than in groups without a glide path (P < 0.05). All instruments were associated with apical extrusion of debris. Creating a glide path prior to canal instrumentation reduced the amount of apically extruded debris in curved canals. © 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  2. Golgi sorting regulates organization and activity of GPI-proteins at apical membranes

    PubMed Central

    Tivodar, Simona; Formiggini, Fabio; Ossato, Giulia; Gratton, Enrico; Tramier, Marc; Coppey-Moisan, Maïté; Zurzolo, Chiara

    2014-01-01

    Here, we combined classical biochemistry with novel biophysical approaches to study with high spatial and temporal resolution the organization of GPI-anchored proteins (GPI-APs) at the plasma membrane of polarized epithelial cells. We show that in polarized MDCK cells, following sorting in the Golgi, each GPI-AP reaches the apical surface in homo-clusters. Golgi-derived homo-clusters are required for their subsequent plasma membrane organization into cholesterol-dependent hetero-clusters. By contrast, in non-polarized MDCK cells GPI-APs are delivered to the surface as monomers in an unpolarized manner and are not able to form hetero-clusters. We further demonstrate that this GPI-AP organization is regulated by the content of cholesterol in the Golgi apparatus and is required to maintain the functional state of the protein at the apical membrane. Thus, different from fibroblasts, in polarized epithelial cells a selective cholesterol-dependent sorting mechanism in the Golgi regulates both the organization and the function of GPI-APs at the apical surface. PMID:24681536

  3. Cryopreservation of redwood (Sequoia sempervirens) in vitro buds using vitrification-based techniques.

    PubMed

    Ozudogru, E A; Kirdok, E; Kaya, E; Capuana, M; Benelli, C; Engelmann, E

    2011-01-01

    In this study, the efficiency of three vitrification-based cryopreservation techniques, i.e. vitrification, encapsulation-vitrification and droplet-vitrification were compared for cryopreserving Sequoia sempervirens apical and basal buds sampled from in vitro shoot cultures. The effect of cold-hardening of mother-plants and of bud culture medium and sucrose preculture was also investigated. Culture of apical and basal buds sampled from cold-hardened mother-plants on Quoirin and Lepoivre medium with activated charcoal had a positive effect on regrowth. Only droplet-vitrification ensured survival and regrowth after cryopreservation. After cryopreservation, regeneration of apical buds was possible for PVS2 exposure durations between 90 and 180 min but it remained low, with a maximum of 18 percent after 135 min treatment. With basal buds, regeneration after cryopreservation was possible over a larger range of PVS2 treatment durations, between 30 and 180 min. The highest regeneration percentage was slightly higher (22 percent) than that measured with apical buds, and was also achieved after 135 min PVS2 exposure.

  4. The V0-ATPase mediates apical secretion of exosomes containing Hedgehog-related proteins in Caenorhabditis elegans

    PubMed Central

    Liégeois, Samuel; Benedetto, Alexandre; Garnier, Jean-Marie; Schwab, Yannick; Labouesse, Michel

    2006-01-01

    Polarized intracellular trafficking in epithelia is critical in development, immunity, and physiology to deliver morphogens, defensins, or ion pumps to the appropriate membrane domain. The mechanisms that control apical trafficking remain poorly defined. Using Caenorhabditis elegans, we characterize a novel apical secretion pathway involving multivesicularbodies and the release of exosomes at the apical plasma membrane. By means of two different genetic approaches, we show that the membrane-bound V0 sector of the vacuolar H+-ATPase (V-ATPase) acts in this pathway, independent of its contribution to the V-ATPase proton pump activity. Specifically, we identified mutations in the V0 “a” subunit VHA-5 that affect either the V0-specific function or the V0+V1 function of the V-ATPase. These mutations allowed us to establish that the V0 sector mediates secretion of Hedgehog-related proteins. Our data raise the possibility that the V0 sector mediates exosome and morphogen release in mammals. PMID:16785323

  5. The apical scaffold big bang binds to spectrins and regulates the growth of Drosophila melanogaster wing discs.

    PubMed

    Forest, Elodie; Logeay, Rémi; Géminard, Charles; Kantar, Diala; Frayssinoux, Florence; Heron-Milhavet, Lisa; Djiane, Alexandre

    2018-03-05

    During development, cell numbers are tightly regulated, ensuring that tissues and organs reach their correct size and shape. Recent evidence has highlighted the intricate connections between the cytoskeleton and the regulation of the key growth control Hippo pathway. Looking for apical scaffolds regulating tissue growth, we describe that Drosophila melanogaster big bang (Bbg), a poorly characterized multi-PDZ scaffold, controls epithelial tissue growth without affecting epithelial polarity and architecture. bbg -mutant tissues are smaller, with fewer cells that are less apically constricted than normal. We show that Bbg binds to and colocalizes tightly with the β-heavy-Spectrin/Kst subunit at the apical cortex and promotes Yki activity, F-actin enrichment, and the phosphorylation of the myosin II regulatory light chain Spaghetti squash. We propose a model in which the spectrin cytoskeleton recruits Bbg to the cortex, where Bbg promotes actomyosin contractility to regulate epithelial tissue growth. © 2018 Forest et al.

  6. An improved model for teaching use of electronic apex locators.

    PubMed

    Tchorz, J P; Hellwig, E; Altenburger, M J

    2012-04-01

    To develop a simple, practical and inexpensive model, which enables the use of electronic apex locators (EALs) during pre-clinical and continuing education. Extracted teeth were placed in a mould and embedded in acrylic resin. The resin was applied in two consecutive steps to form a cavity around the root apices. A closable plastic tube serves as a valve, and a steel wire connects to the EAL. With its semi-closed reservoir for conductive fluids surrounding the root apices, the new model enables working length measurements of root canals using EALs. The model simulates the clinical situation for endodontic teaching purposes, as it allows working length determination of root canals as recommended. The measuring results of the EAL can be verified by radiography. At the same time, the roots are not directly visible and accessible to the user, allowing a precise evaluation and grading of the treatment. © 2011 International Endodontic Journal.

  7. GLUT2 Accumulation in Enterocyte Apical and Intracellular Membranes

    PubMed Central

    Ait-Omar, Amal; Monteiro-Sepulveda, Milena; Poitou, Christine; Le Gall, Maude; Cotillard, Aurélie; Gilet, Jules; Garbin, Kevin; Houllier, Anne; Château, Danièle; Lacombe, Amélie; Veyrie, Nicolas; Hugol, Danielle; Tordjman, Joan; Magnan, Christophe; Serradas, Patricia; Clément, Karine; Leturque, Armelle; Brot-Laroche, Edith

    2011-01-01

    OBJECTIVE In healthy rodents, intestinal sugar absorption in response to sugar-rich meals and insulin is regulated by GLUT2 in enterocyte plasma membranes. Loss of insulin action maintains apical GLUT2 location. In human enterocytes, apical GLUT2 location has not been reported but may be revealed under conditions of insulin resistance. RESEARCH DESIGN AND METHODS Subcellular location of GLUT2 in jejunal enterocytes was analyzed by confocal and electron microscopy imaging and Western blot in 62 well-phenotyped morbidly obese subjects and 7 lean human subjects. GLUT2 locations were assayed in ob/ob and ob/+ mice receiving oral metformin or in high-fat low-carbohydrate diet–fed C57Bl/6 mice. Glucose absorption and secretion were respectively estimated by oral glucose tolerance test and secretion of [U-14C]-3-O-methyl glucose into lumen. RESULTS In human enterocytes, GLUT2 was consistently located in basolateral membranes. Apical GLUT2 location was absent in lean subjects but was observed in 76% of obese subjects and correlated with insulin resistance and glycemia. In addition, intracellular accumulation of GLUT2 with early endosome antigen 1 (EEA1) was associated with reduced MGAT4a activity (glycosylation) in 39% of obese subjects on a low-carbohydrate/high-fat diet. Mice on a low-carbohydrate/high-fat diet for 12 months also exhibited endosomal GLUT2 accumulation and reduced glucose absorption. In ob/ob mice, metformin promoted apical GLUT2 and improved glucose homeostasis. Apical GLUT2 in fasting hyperglycemic ob/ob mice tripled glucose release into intestinal lumen. CONCLUSIONS In morbidly obese insulin-resistant subjects, GLUT2 was accumulated in apical and/or endosomal membranes of enterocytes. Functionally, apical GLUT2 favored and endosomal GLUT2 reduced glucose transepithelial exchanges. Thus, altered GLUT2 locations in enterocytes are a sign of intestinal adaptations to human metabolic pathology. PMID:21852673

  8. Detection of zinc translocation into apical dendrite of CA1 pyramidal neuron after electrical stimulation.

    PubMed

    Suh, Sang Won

    2009-02-15

    Translocation of the endogenous cation zinc from presynaptic terminals to postsynaptic neurons after brain insult has been implicated as a potential neurotoxic event. Several studies have previously demonstrated that a brief electrical stimulation is sufficient to induce the translocation of zinc from presynaptic vesicles into the cytoplasm (soma) of postsynaptic neurons. In the present work I have extended those findings in three ways: (i) providing evidence that zinc translocation occurs into apical dendrites, (ii) presenting data that there is an apparent translocation into apical dendrites when only a zinc-containing synaptic input is stimulated, and (iii) presenting data that there is no zinc translocation into apical dendrite of ZnT3 KO mice following electrical stimulation. Hippocampal slices were preloaded with the "trappable" zinc fluorescent probe, Newport Green. After washout, a single apical dendrite in the stratum radiatum of hippocampal CA1 area was selected and focused on. Burst stimulation (100Hz, 500microA, 0.2ms, monopolar) was delivered to either the adjacent Schaffer-collateral inputs (zinc-containing) or to the adjacent temporo-ammonic inputs (zinc-free) to the CA1 dendrites. Stimulation of the Schaffer collaterals increased the dendritic fluorescence, which was blocked by TTX, low-Ca medium, or the extracellular zinc chelator, CaEDTA. Stimulation of the temporo-ammonic pathway caused no significant rise in the fluorescence. Genetic depletion of vesicular zinc by ZnT3 KO showed no stimulation-induced apical dendrite zinc rise. The present study provides evidence that synaptically released zinc translocates into postsynaptic neurons through the apical dendrites of CA1 pyramidal neurons during physiological synaptic activity.

  9. Effects of loxoprofen on the apical root resorption during orthodontic tooth movement in rats.

    PubMed

    Yamamoto, Taeko; Kaku, Masato; Sumi, Hiromi; Yashima, Yuka; Izumino, Jin; Tanimoto, Kotaro

    2018-01-01

    Studies have revealed that severe apical root resorption during tooth movement is caused by the noninfective inflammatory reaction of apical root tissues. We hypothesized that loxoprofen can suppress apical root resorption during tooth movement. Cyclic tensile force (CTF) of 10 kPa was applied to the human pulp cells for 48 hours by the Flexcell Strain Unit. Loxoprofen (10 and 100 μM) was added to the culture cells, and expression of cyclooxygenase (COX)-1, COX-2, interleukin (IL)-1β, receptor activator of nuclear factor kappa-B ligand (RANKL), tumor necrosis factor (TNF)-α, and macrophage colony-stimulating factor (M-CSF) were examined. To determine the effects of loxoprofen sodium on apical root reabsorption during tooth movement, the upper first molars of 7-week-old rats were subjected to mesial movement by 10g force for 30 days with or without the oral administration of loxoprofen. Gene expression and protein concentration of COX-1, COX-2, IL-1β, TNF-α, RANKL and M-CSF were significantly higher in the CTF group than in the control group. However, these levels were decreased by loxoprofen administration. After orthodontic tooth movement, the expression of IL-1β, TNF-α, RANKL and M-CSF decreased in the loxoprofen group than in the control group by immunohistochemical staining. In comparison to control group, less number of odontoclasts and a decrease in the amount of apical root resorption was observed in the loxoprofen group. Many osteoclasts became visible on the pressure side of the alveolar bone in the both groups, and the amount of tooth movement did not show a significant difference. These findings demonstrate that severe apical root resorption may be suppressed by loxoprofen administration, without a disturbance of tooth movement.

  10. Effects of loxoprofen on the apical root resorption during orthodontic tooth movement in rats

    PubMed Central

    Sumi, Hiromi; Yashima, Yuka; Izumino, Jin

    2018-01-01

    Studies have revealed that severe apical root resorption during tooth movement is caused by the noninfective inflammatory reaction of apical root tissues. We hypothesized that loxoprofen can suppress apical root resorption during tooth movement. Cyclic tensile force (CTF) of 10 kPa was applied to the human pulp cells for 48 hours by the Flexcell Strain Unit. Loxoprofen (10 and 100 μM) was added to the culture cells, and expression of cyclooxygenase (COX)-1, COX-2, interleukin (IL)-1β, receptor activator of nuclear factor kappa-B ligand (RANKL), tumor necrosis factor (TNF)-α, and macrophage colony-stimulating factor (M-CSF) were examined. To determine the effects of loxoprofen sodium on apical root reabsorption during tooth movement, the upper first molars of 7-week-old rats were subjected to mesial movement by 10g force for 30 days with or without the oral administration of loxoprofen. Gene expression and protein concentration of COX-1, COX-2, IL-1β, TNF-α, RANKL and M-CSF were significantly higher in the CTF group than in the control group. However, these levels were decreased by loxoprofen administration. After orthodontic tooth movement, the expression of IL-1β, TNF-α, RANKL and M-CSF decreased in the loxoprofen group than in the control group by immunohistochemical staining. In comparison to control group, less number of odontoclasts and a decrease in the amount of apical root resorption was observed in the loxoprofen group. Many osteoclasts became visible on the pressure side of the alveolar bone in the both groups, and the amount of tooth movement did not show a significant difference. These findings demonstrate that severe apical root resorption may be suppressed by loxoprofen administration, without a disturbance of tooth movement. PMID:29694352

  11. Isolation and identification of Enterococcus faecalis from necrotic root canals using multiplex PCR.

    PubMed

    Mahmoudpour, Ali; Rahimi, Saeed; Sina, Mahmood; Soroush, Mohammad H; Shahi, Shahriar; Shahisa, Shahriar; Asl-Aminabadi, Naser

    2007-09-01

    This study was designed to survey the incidence of Enterococcus faecalis infection in symptomatic and asymptomatic root canals of necrotic teeth using PCR and to isolate the bacterium for further screening. Sixty patients categorized according to their clinical symptoms were used for sampling by insertion of paper points into the root canals and absorbing all the fluids present within them. The samples were incubated in 1.0 ml 2xYT (containing 16 g bacto tryptone, 10 g yeast extract and 5.0 g NaCl per liter) for 24 h at 37 degrees C without aeration prior to multiplex PCR analysis. To assist the isolation of E. faecalis, sub-samples were further grown in the same medium supplemented with 6.5% NaCl and back-inoculated into bile esculin. Using multiple cultivation-dependent and PCR analyses, 6 cases (10%) of E. faecalis were identified. Four isolates were obtained from asymptomatic cases of chronic apical periodontitis, and the other two were associated with phoenix abscess and acute apical abscess, respectively. No E. faecalis infection was found in 5 patients with acute apical periodontitis or in 9 with chronic suppurative periodontitis. Our results indicate that there is no significant difference in the incidence of E. faecalis between symptomatic and asymptomatic necrotic dental root canals (P > 0.05).

  12. Comparison of the EndoVac system and conventional needle irrigation on removal of the smear layer in primary molar root canals.

    PubMed

    Buldur, B; Kapdan, A

    2017-09-01

    This study aimed to compare the EndoVac system and conventional needle irrigation in removing smear layer (SR) from primary molar root canals. Fifty extracted human primary second molar roots were instrumented up to an apical size of 0.04/35 and randomly divided into two main groups; Group 1: EndoVac system (n = 25) and Group 2: Conventional needle irrigation (n = 25) and three subgroups (a) NaOCl + ethylenediaminetetraacetic acid (EDTA) (n = 20) (b) ozonated water (OW) + EDTA (n = 20) and (c) saline (control, n = 10). After a standardized final irrigation protocol performed for all teeth, scanning electron microscope images were taken at ×1000 magnification for each thirds of each root canal. Data were analyzed by the weighted kappa, Kruskal-Wallis, and Wilcoxon signed rank tests. EndoVac was more effective than conventional needle in the removal of SR from the apical third of the root canal system (P < 0.05). The OW + EDTA regimen provided similar SR removal compared with NaOCl + EDTA. EndoVac has better performance than conventional needle irrigation in the removal of the SR in the apical thirds of the primary molar root canals. As a final irrigation regimen, the OW + EDTA regimen is as effective as the NaOCl + EDTA regimen.

  13. Apical extrusion of debris and irrigants using ProTaper hand, M-two rotary and WaveOne single file reciprocating system: An ex vivo study.

    PubMed

    Singh, Abhishek; Arunagiri, Doraiswamy; Pushpa, Shankarappa; Sawhny, Asheesh; Misra, Abhinav; Khetan, Kirti

    2015-01-01

    The purpose of this ex vivo study was to evaluate and compare the weight of debris and volume of irrigant extruded apically from teeth using different preparation techniques. Thirty extracted human mandibular premolars with single canals and similar lengths were instrumented using hand ProTaper F2 (25, 0.08; Dentsply Maillefer, Ballaigues, Switzerland), M-two (25, 0.06; VDW, Munich, Germany) and WaveOne Primary (25, 0.08; Dentsply Maillefer, Ballaigues, Switzerland). Debris and irrigant extruded during instrumentation were collected into preweighed Eppendorf tubes. The volume of the irrigant was measured, and then the tubes were stored in an incubator at 70°C for 2 days. The Eppendorf tubes were weighed to obtain the final weight when the extruded debris was included. Three consecutive weights were obtained for each tube. Data were statistically analyzed by one-way analysis of variance and Student's t-test. There were no statistically significant differences among the groups. The WaveOne reciprocating system showed the maximum amount of apical extrusion of debris and irrigant among all the groups. The least amount of debris and irrigant was observed in ProTaper hand instrument (P > 0.05). All instrumentation techniques were associated with debris and irrigant extrusion.

  14. A Descriptive Analysis of Dental Conditions Occurring During Conflicts, Deployments, and Field Training Exercises

    DTIC Science & Technology

    1999-12-01

    Infection of an extraction site (dry socket, localized osteitis). Apical abscess /periapical abscess - Collection of purulent exudate around the area...conditions for deployment data were 5.79% higher than the rate during conflicts. Caries, third molar complications, periapical abscesses , and endodontics are...restorations and fractured teeth. The "Periodontal" category was grouped to include periodontal abscess , acute necrotizing gingivitis (ANUG), and

  15. Investigation of the Efficacy of Passive Ultrasonic Irrigation Versus Irrigation with Reciprocating Activation: An Environmental Scanning Electron Microscopic Study.

    PubMed

    Kato, Augusto Shoji; Cunha, Rodrigo Sanches; da Silveira Bueno, Carlos Eduardo; Pelegrine, Rina Andrea; Fontana, Carlos Eduardo; de Martin, Alexandre Sigrist

    2016-04-01

    The objective of this ex vivo study was to compare the efficacy of passive ultrasonic irrigation (PUI) versus a new activation system using reciprocating motion (EasyClean [EC]; Easy Equipamentos Odontológicos, Belo Horizonte, Brazil) to remove debris from the root canal walls at 6 predetermined apical levels using environmental scanning electron microscopy. Mesiobuccal root canals of 10 mandibular molars were prepared with a 30/.05 final instrument. The specimens were embedded in flasks containing heavy body silicone, cleaved longitudinally, and 6 round indentations were made into the apical region of the buccal half at 1-mm intervals. The same specimens were used to prepare a blank control group (no debris), a negative control group (completely covered by debris), and 2 experimental groups: PUI and irrigation with reciprocating activation. Standardized images of the indentations were obtained under environmental scanning electron microscopy and assessed by 2 examiners. The amount of debris was then classified using a 4-category scoring system. The kappa test was applied to determine interexaminer agreement, whereas the Kruskal-Wallis, Dunn, and Friedman tests were used to compare scores. The EC group had results statistically similar to those of the blank control group for all 6 root levels examined. The PUI group had results statistically similar to those of the negative control group for the 3 most apical levels and similar to those of the blank control group for the 3 most cervical levels. Activating the irrigant with a reciprocating system (EC) promoted more effective debris removal from the more apical regions of the root canal when compared with PUI. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  16. Cemental tear: To know what we have neglected in dental practice.

    PubMed

    Jeng, Po-Yuan; Luzi, Arlinda Luzi; Pitarch, Rocio Marco; Chang, Mei-Chi; Wu, Yu-Hsueh; Jeng, Jiiang-Huei

    2018-04-01

    Cemental tear is a special kind of root surface fracture, contributing to periodontal and periapical breakdown. However, it is a challenge for doctors to diagnose, resulting in delayed or improper treatment. We reviewed the predisposing factors, location, radiographic/clinical characteristics, diagnosis and treatments of cemental tears. From the literature, patients with cemental tear were mainly males, over 60 year-old. Possible predisposing factors include gender, age, tooth type, traumatic occlusal force and vital teeth. Cemental tears were common in upper and lower anterior teeth, single or multiple, and can be present in cervical, middle and apical third of roots. Morphology of cemental tears can be either piece-shaped or U-shaped. Clinically, cemental tear shows a unitary periodontal pocket and signs/symptoms mimicking localized periodontitis, apical periodontitis and vertical root fractures. Treatment of cemental tears include scaling, root planning, root canal treatment, periodontal/periapical surgery, guided tissue regeneration, bone grafting, and intentional replantation. Recurrence of cemental tear is possible especially when the fracture involves root apex. Extraction is recommended for teeth with poor prognosis. In conclusion, cemental tears can involve both periodontal and periapical area. Dentists should understand the predisposing factors and clinical features of cemental tears for early diagnosis/treatment to prevent bone loss/tooth extraction. Copyright © 2017. Published by Elsevier B.V.

  17. [Effects of lipopolysaccharides extracted from Porphyromonas endodontalis on the expression of IL-1beta mRNA and IL-6 mRNA in osteoblasts].

    PubMed

    Yang, Di; Li, Ren; Qiu, Li-Hong; Li, Chen

    2009-04-01

    To quantify the IL-1 beta mRNA and IL-6 mRNA expression induced by lipopolysaccharides (LPS)extracted from Porphyromonas endodontalis(P.e) in osteoblasts, and to relate P.e-LPS to bone absorption pathogenesis in lesions of chronical apical periodontitis. MG63 was treated with different concentrations of P.e-LPS(0-50 microg/mL) for different hours(0-24h). The expression of IL-1 beta mRNA and IL-6 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR).Statistical analysis was performed using one- way ANOVA and Dunnett t test with SPSS11.0 software package. The level of IL-1 beta mRNA and IL-6 mRNA increased significantly after treatment with P.e-LPS at more than 5 microg/mL (P<0.01)and for more than 1 hour (P<0.01), which indicated that P.e-LPS induced osteoblasts to express IL-1 beta mRNA and IL-6 mRNA in dose and time dependent manners. P.e-LPS may promote bone resorption in lesions of chronical apical periodontitis by inducing IL-1 beta mRNA and IL-6 mRNA expression in osteoblasts.

  18. Arctigenin from Fructus Arctii (Seed of Burdock) Reinforces Intestinal Barrier Function in Caco-2 Cell Monolayers

    PubMed Central

    Shin, Hee Soon; Jung, Sun Young; Back, Su Yeon; Do, Jeong-Ryong; Shon, Dong-Hwa

    2015-01-01

    Fructus Arctii is used as a traditional herbal medicine to treat inflammatory diseases in oriental countries. This study aimed to investigate effect of F. Arctii extract on intestinal barrier function in human intestinal epithelial Caco-2 cells and to reveal the active component of F. Arctii. We measured transepithelial electrical resistance (TEER) value (as an index of barrier function) and ovalbumin (OVA) permeation (as an index of permeability) to observe the changes of intestinal barrier function. The treatment of F. Arctii increased TEER value and decreased OVA influx on Caco-2 cell monolayers. Furthermore, we found that arctigenin as an active component of F. Arctii increased TEER value and reduced permeability of OVA from apical to the basolateral side but not arctiin. In the present study, we revealed that F. Arctii could enhance intestinal barrier function, and its active component was an arctigenin on the functionality. We expect that the arctigenin from F. Arctii could contribute to prevention of inflammatory, allergic, and infectious diseases by reinforcing intestinal barrier function. PMID:26550018

  19. Spectrophotometric determination of irrigant extrusion using passive ultrasonic irrigation, EndoActivator, or syringe irrigation.

    PubMed

    Rodríguez-Figueroa, Carolina; McClanahan, Scott B; Bowles, Walter R

    2014-10-01

    Sodium hypochlorite (NaOCl) irrigation is critical to endodontic success, and several new methods have been developed to improve irrigation efficacy (eg, passive ultrasonic irrigation [PUI] and EndoActivator [EA]). Using a novel spectrophotometric method, this study evaluated NaOCl irrigant extrusion during canal irrigation. One hundred fourteen single-rooted extracted teeth were decoronated to leave 15 mm of the root length for each tooth. Cleaning and shaping of the teeth were completed using standardized hand and rotary instrumentation to an apical file size #40/0.04 taper. Roots were sealed (not apex), and 54 straight roots (n = 18/group) and 60 curved roots (>20° curvature, n = 20/group) were included. Teeth were irrigated with 5.25% NaOCl by 1 of 3 methods: passive irrigation with needle, PUI, or EA irrigation. Extrusion of NaOCl was evaluated using a pH indicator and a spectrophotometer. Standard curves were prepared with known amounts of irrigant to quantify amounts in unknown samples. Irrigant extrusion was minimal with all methods, with most teeth showing no NaOCl extrusion in straight or curved roots. Minor NaOCl extrusion (1-3 μL) in straight roots or curved roots occurred in 10%-11% of teeth in all 3 irrigant methods. Two teeth in both the syringe irrigation and the EA group extruded 3-10 μL of NaOCl. The spectrophotometric method used in this study proved to be very sensitive while providing quantification of the irrigant levels extruded. Using the PUI or EA tip to within 1 mm of the working length appears to be fairly safe, but apical anatomy can vary in teeth to allow extrusion of irrigant. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  20. Efficient extraction of vaccines formulated in aluminum hydroxide gel by including surfactants in the extraction buffer

    PubMed Central

    Zhu, Daming; Huang, Shuhui; McClellan, Holly; Dai, Weili; Syed, Najam R; Gebregeorgis, Elizabeth; Mullen, Gregory E. D.; Long, Carole; Martin, Laura B.; Narum, David; Duffy, Patrick; Miller, Louis H.; Saul, Allan

    2011-01-01

    Efficient antigen extraction from vaccines formulated on aluminum hydroxide gels is a critical step for the evaluation of the quality of vaccines following formulation. It has been shown in our laboratory that the efficiency of antigen extraction from vaccines formulated on Alhydrogel decreased significantly with increased storage time. To increase antigen extraction efficiency, the present study determined the effect of surfactants on antigen recovery from vaccine formulations. The Plasmodium falciparum apical membrane antigen 1 (AMA1) formulated on Alhydrogel and stored at 2-8 °C for three years was used as a model in this study. The AMA1 on Alhydrogel was extracted in the presence or absence of 30 mM sodium dodecyl sulfate (SDS) or 20 mM cetylpyridinium chloride in the extraction buffer (0.60 M citrate, 0.55 M phosphate, pH 8.5) using our standard antigen extraction protocols. Extracted AMA1 antigen was analyzed by 4-20% Tris-glycine SDS-PAGE followed by silver staining or western blotting. The results showed that inclusion of SDS or cetylpyridinium chloride in extraction buffer increased the antigen recovery dramatically and can be used for efficient characterization of Alhydrogel vaccines. PMID:22107848

  1. Nerve growth factor reduces amiloride‐sensitive Na+ transport in human airway epithelial cells

    PubMed Central

    Shimko, Michael J.; Zaccone, Eric J.; Thompson, Janet A.; Schwegler‐Berry, Diane; Kashon, Michael L.; Fedan, Jeffrey S.

    2014-01-01

    Abstract Nerve growth factor (NGF) is overexpressed in patients with inflammatory lung diseases, including virus infections. Airway surface liquid (ASL), which is regulated by epithelial cell ion transport, is essential for normal lung function. No information is available regarding the effect of NGF on ion transport of airway epithelium. To investigate whether NGF can affect ion transport, human primary air‐interface cultured epithelial cells were placed in Ussing chambers to obtain transepithelial voltage (−7.1 ± 3.4 mV), short‐circuit current (Isc, 5.9 ± 1.0 μA), and transepithelial resistance (750 Ω·cm2), and to measure responses to ion transport inhibitors. Amiloride (apical, 3.5 × 10−5 mol/L) decreased Isc by 55.3%. Apically applied NGF (1 ng/mL) reduced Isc by 5.3% in 5 min; basolaterally applied NGF had no effect. The response to amiloride was reduced (41.6%) in the presence of NGF. K‐252a (10 nmol/L, apical) did not itself affect Na+ transport, but it attenuated the NGF‐induced reduction in Na+ transport, indicating the participation of the trkA receptor in the NGF‐induced reduction in Na+ transport. PD‐98059 (30 μmol/L, apical and basolateral) did not itself affect Na+ transport, but attenuated the NGF‐induced reduction in Na+ transport, indicating that trkA activated the Erk 1/2 signaling cascade. NGF stimulated phosphorylation of Erk 1/2 and the β‐subunit of ENaC. K‐252a and PD‐98059 inhibited these responses. NGF had no effect on Isc in the presence of apical nystatin (50 μmol/L). These results indicate that NGF inhibits Na+ transport through a trkA‐Erk 1/2‐activated signaling pathway linked to ENaC phosphorylation. PMID:25347857

  2. Theory of electric resonance in the neocortical apical dendrite.

    PubMed

    Kasevich, Ray S; LaBerge, David

    2011-01-01

    Pyramidal neurons of the neocortex display a wide range of synchronous EEG rhythms, which arise from electric activity along the apical dendrites of neocortical pyramidal neurons. Here we present a theoretical description of oscillation frequency profiles along apical dendrites which exhibit resonance frequencies in the range of 10 to 100 Hz. The apical dendrite is modeled as a leaky coaxial cable coated with a dielectric, in which a series of compartments act as coupled electric circuits that gradually narrow the resonance profile. The tuning of the peak frequency is assumed to be controlled by the average amplitude of voltage-gated outward currents, which in turn are regulated by the subthreshold noise in the thousands of synaptic spines that are continuously bombarded by local circuits. The results of simulations confirmed the ability of the model both to tune the peak frequency in the 10-100 Hz range and to gradually narrow the resonance profile. Considerable additional narrowing of the resonance profile is provided by repeated looping through the apical dendrite via the corticothalamocortical circuit, which reduced the width of each resonance curve (at half-maximum) to approximately 1 Hz. Synaptic noise in the neural circuit is discussed in relation to the ways it can influence the narrowing process.

  3. Theory of Electric Resonance in the Neocortical Apical Dendrite

    PubMed Central

    Kasevich, Ray S.; LaBerge, David

    2011-01-01

    Pyramidal neurons of the neocortex display a wide range of synchronous EEG rhythms, which arise from electric activity along the apical dendrites of neocortical pyramidal neurons. Here we present a theoretical description of oscillation frequency profiles along apical dendrites which exhibit resonance frequencies in the range of 10 to 100 Hz. The apical dendrite is modeled as a leaky coaxial cable coated with a dielectric, in which a series of compartments act as coupled electric circuits that gradually narrow the resonance profile. The tuning of the peak frequency is assumed to be controlled by the average amplitude of voltage-gated outward currents, which in turn are regulated by the subthreshold noise in the thousands of synaptic spines that are continuously bombarded by local circuits. The results of simulations confirmed the ability of the model both to tune the peak frequency in the 10–100 Hz range and to gradually narrow the resonance profile. Considerable additional narrowing of the resonance profile is provided by repeated looping through the apical dendrite via the corticothalamocortical circuit, which reduced the width of each resonance curve (at half-maximum) to approximately 1 Hz. Synaptic noise in the neural circuit is discussed in relation to the ways it can influence the narrowing process. PMID:21853129

  4. Chemical form of selenium affects its uptake, transport, and glutathione peroxidase activity in the human intestinal Caco-2 cell model.

    PubMed

    Zeng, Huawei; Jackson, Matthew I; Cheng, Wen-Hsing; Combs, Gerald F

    2011-11-01

    Determining the effect of selenium (Se) chemical form on uptake, transport, and glutathione peroxidase activity in human intestinal cells is critical to assess Se bioavailability at nutritional doses. In this study, we found that two sources of L-selenomethionine (SeMet) and Se-enriched yeast each increased intracellular Se content more effectively than selenite or methylselenocysteine (SeMSC) in the human intestinal Caco-2 cell model. Interestingly, SeMSC, SeMet, and digested Se-enriched yeast were transported at comparable efficacy from the apical to basolateral sides, each being about 3-fold that of selenite. In addition, these forms of Se, whether before or after traversing from apical side to basolateral side, did not change the potential to support glutathione peroxidase (GPx) activity. Although selenoprotein P has been postulated to be a key Se transport protein, its intracellular expression did not differ when selenite, SeMSC, SeMet, or digested Se-enriched yeast was added to serum-contained media. Taken together, our data show, for the first time, that the chemical form of Se at nutritional doses can affect the absorptive (apical to basolateral side) efficacy and retention of Se by intestinal cells; but that, these effects are not directly correlated to the potential to support GPx activity.

  5. Stochastic phase of ventral furrow formation in the Drosophila embryo: cellular constriction chains, mechanical feedback, and robustness

    NASA Astrophysics Data System (ADS)

    Blawzdziewicz, Jerzy; Gao, Guo-Jie J.; Holcomb, Michael C.; Thomas, Jeffrey H.

    The key process giving rise to ventral furrow formation (VFF) in Drosophila embryo is apical constriction of cells in the ventral region. The constriction produces negative spontaneous curvature of the cell layer. During the initial slower phase of VFF approximately 40% of cells constrict in a seemingly random order. We show that this initial phase of VFF does not depend on random uncorrelated events. Instead, constricted cell apices form well-defined correlated structures, i.e., cellular constriction chains (CCCs), indicative of strong spatial and directional correlations between the constriction events. We argue that this chain formation is a signature of mechanical signaling that coordinates apical constrictions through tensile stress. To gain insights into the mechanisms involved in this correlated constriction process, we propose an active granular fluid (AGF) model which considers a tissue as a collection of mechanically active, stress-responsive objects. Our AGF molecular dynamics simulations show that cell constriction sensitivity to tensile stress results in formation of CCCs whereas compressive-stress sensitivity leads to compact constricted cell clusters; the CCCs, which can penetrate less-active regions, increase the robustness of the VFF process.

  6. Glycolysis regulates pollen tube polarity via Rho GTPase signaling

    PubMed Central

    Chen, Wei; Gong, Pingping; Guo, Jingzhe; Li, Hui; Li, Ruizi; Xing, Weiman; Yang, Zhenbiao

    2018-01-01

    As a universal energy generation pathway utilizing carbon metabolism, glycolysis plays an important housekeeping role in all organisms. Pollen tubes expand rapidly via a mechanism of polarized growth, known as tip growth, to deliver sperm for fertilization. Here, we report a novel and surprising role of glycolysis in the regulation of growth polarity in Arabidopsis pollen tubes via impingement of Rho GTPase-dependent signaling. We identified a cytosolic phosphoglycerate kinase (pgkc-1) mutant with accelerated pollen germination and compromised pollen tube growth polarity. pgkc-1 mutation greatly diminished apical exocytic vesicular distribution of REN1 RopGAP (Rop GTPase activating protein), leading to ROP1 hyper-activation at the apical plasma membrane. Consequently, pgkc-1 pollen tubes contained higher amounts of exocytic vesicles and actin microfilaments in the apical region, and showed reduced sensitivity to Brefeldin A and Latrunculin B, respectively. While inhibition of mitochondrial respiration could not explain the pgkc-1 phenotype, the glycolytic activity is indeed required for PGKc function in pollen tubes. Moreover, the pgkc-1 pollen tube phenotype was mimicked by the inhibition of another glycolytic enzyme. These findings highlight an unconventional regulatory function for a housekeeping metabolic pathway in the spatial control of a fundamental cellular process. PMID:29702701

  7. Dietary starch breakdown product sensing mobilizes and apically activates α-glucosidases in small intestinal enterocytes.

    PubMed

    Chegeni, Mohammad; Amiri, Mahdi; Nichols, Buford L; Naim, Hassan Y; Hamaker, Bruce R

    2018-02-20

    Dietary starch is finally converted to glucose for absorption by the small intestine mucosal α-glucosidases (sucrase-isomaltase [SI] and maltase-glucoamylase), and control of this process has health implications. Here, the molecular mechanisms were analyzed associated with starch-triggered maturation and transport of SI. Biosynthetic pulse-chase in Caco-2 cells revealed that the high MW SI species (265 kDa) induced by maltose (an α-amylase starch digestion product) had a higher rate of early trafficking and maturation compared with a glucose-induced SI (245 kDa). The maltose-induced SI was found to have higher affinity to lipid rafts, which are associated with enhanced targeting to the apical membrane and higher activity. Accordingly, in situ maltose-hydrolyzing action was enhanced in the maltose-treated cells. Thus, starch digestion products at the luminal surface of small intestinal enterocytes are sensed and accelerate the intracellular processing of SI to enhance starch digestion capacity in the intestinal lumen.-Chegeni, M., Amiri, M., Nichols, B. L., Naim, H. Y., Hamaker, B. R. Dietary starch breakdown product sensing mobilizes and apically activates α-glucosidases in small intestinal enterocytes.

  8. Single-channel analysis of functional epithelial sodium channel (ENaC) stability at the apical membrane of A6 distal kidney cells

    PubMed Central

    Yu, Ling; Helms, My N.; Yue, Qiang; Eaton, Douglas C.

    2008-01-01

    Epithelial sodium channels (ENaC) play an essential role in maintaining total body fluid and electrolyte homeostasis. As such, abnormal expression of ENaC at the cell surface is linked to several important human diseases. Although the stability of ENaC subunits has been extensively studied by protein biochemical analysis, the half-life of the functional channel in the apical membrane remains controversial. Because the functional stability of the multisubunit channel may be more physiologically relevant than the stability of individual subunit proteins, we performed studies of functional ENaC channels using A6 epithelial cells, a Xenopus laevis distal nephron cell line. We recorded single-channel activity in over 400 cells with the translation blockers cycloheximide (CHX) or puromycin, as well as the intracellular protein trafficking inhibitors brefeldin A (BFA) or nocodazole. Our cell-attached, single-channel recordings allow us to quantify the channel density in the apical membrane, as well as to determine channel open probability (Po) from control (untreated) cells and from cells at different times of drug treatment. The data suggest that the half-life of ENaC channels is ∼3.5 h following puromycin, BFA, and nocodazole treatment. Furthermore, these three drugs had no significant effect on the Po of ENaC for at least 6 h after exposure. A decrease in apical channel number and Po was observed following 2 h of CHX inhibition of protein synthesis, and the apparent channel half-life was closer to 1.5 h following CHX treatment. Treatment of cells with the translation inhibitors does not alter the expression of the protease furin, and therefore changes in protease activity cannot explain changes in ENaC Po. Confocal images show that BFA and nocodazole both disrupt most of the Golgi apparatus after 1-h exposure. In cells with the Golgi totally disrupted by overnight exposure to BFA, 20% of apical ENaC channels remained functional. This result suggests that ENaC is delivered to the apical membrane via a pathway that might bypass the Golgi vesicular trafficking pathway, or that there might be two pools of channels with markedly different half-lives in the apical membrane. PMID:18784262

  9. Single-channel analysis of functional epithelial sodium channel (ENaC) stability at the apical membrane of A6 distal kidney cells.

    PubMed

    Yu, Ling; Helms, My N; Yue, Qiang; Eaton, Douglas C

    2008-11-01

    Epithelial sodium channels (ENaC) play an essential role in maintaining total body fluid and electrolyte homeostasis. As such, abnormal expression of ENaC at the cell surface is linked to several important human diseases. Although the stability of ENaC subunits has been extensively studied by protein biochemical analysis, the half-life of the functional channel in the apical membrane remains controversial. Because the functional stability of the multisubunit channel may be more physiologically relevant than the stability of individual subunit proteins, we performed studies of functional ENaC channels using A6 epithelial cells, a Xenopus laevis distal nephron cell line. We recorded single-channel activity in over 400 cells with the translation blockers cycloheximide (CHX) or puromycin, as well as the intracellular protein trafficking inhibitors brefeldin A (BFA) or nocodazole. Our cell-attached, single-channel recordings allow us to quantify the channel density in the apical membrane, as well as to determine channel open probability (Po) from control (untreated) cells and from cells at different times of drug treatment. The data suggest that the half-life of ENaC channels is approximately 3.5 h following puromycin, BFA, and nocodazole treatment. Furthermore, these three drugs had no significant effect on the Po of ENaC for at least 6 h after exposure. A decrease in apical channel number and Po was observed following 2 h of CHX inhibition of protein synthesis, and the apparent channel half-life was closer to 1.5 h following CHX treatment. Treatment of cells with the translation inhibitors does not alter the expression of the protease furin, and therefore changes in protease activity cannot explain changes in ENaC Po. Confocal images show that BFA and nocodazole both disrupt most of the Golgi apparatus after 1-h exposure. In cells with the Golgi totally disrupted by overnight exposure to BFA, 20% of apical ENaC channels remained functional. This result suggests that ENaC is delivered to the apical membrane via a pathway that might bypass the Golgi vesicular trafficking pathway, or that there might be two pools of channels with markedly different half-lives in the apical membrane.

  10. A comparative study of the debridement efficacy and apical extrusion of dynamic and passive root canal irrigation systems

    PubMed Central

    2014-01-01

    Background Root canal irrigation carries a risk of extrusion of irrigant into the periapical tissues which can be associated with pain, swelling, and tissue damage. Studies have shown less extrusion with sonic or apical negative pressure devices compared with syringe and side-port needle or passive ultrasonic irrigation with continuous irrigant flow. This study aimed to evaluate the effectiveness of the EndoVac irrigation system, regarding 1) debris removal and 2) the control of apically extruded irrigating solution. Methods Fifty extracted human single-rooted teeth were used in this study. The teeth were then randomly divided into three experimental groups according to the type of irrigation used and one control group. In group 1, irrigation was performed using the EndoVac irrigation system. In group 2, irrigation was performed using a 30-gauge, tip-vented irrigation needle. In group 3, irrigation was performed using a 30-gauge, side-vented irrigation needle. The control group received instrumentation with no irrigation to serve as a control for cleaning efficiency. Root canal instrumentation was performed using the Profile NiTi rotary system with a crown-down technique. All of the experimental teeth were irrigated with the same amount of 5.25% sodium hypochlorite. The amount of extruded irrigating solution was then measured by subtracting the post-instrumentation weight from the pre-instrumentation weight using an electronic balance. The cleanliness of debris removal was evaluated using scanning electron microscopy. Results EndoVac irrigation had the least amount of extrusion followed by the side-vented and tip-vented method. The difference between the groups was statistically significant (P <0.01). As for the cleaning results, the debris collection in the EndoVac and tip-vented groups was the least in the apical third. In the control and the side-vented groups, the debris was the greatest in the apical third, but this difference was not significant among the three experimental groups. Conclusions The EndoVac irrigation system extruded significantly less irrigant solution than either needle irrigation system. Debris collection was the least in the apical third for the EndoVac irrigation system. No significant difference was found in the cleaning efficiency among the three irrigation systems. PMID:24512441

  11. An In-Vitro Evaluation and Comparison of Apical Sealing Ability of Three Different Obturation Technique - Lateral Condensation, Obtura II, and Thermafil.

    PubMed

    Emmanuel, Samson; Shantaram, Kulkarni; Sushil, Kumar C; Manoj, Likhitkar

    2013-04-01

    Success of non-surgical root canal treatment is predicted by meticulous cleaning and shaping of the root canal system, three-dimensional obturation and a well-fitting "leakage-free" coronal restoration. The techniques of obturation that are available have their own relative position in the historical development of filling techniques. Over the years, pitfalls with one technique have often led to the development of newer methods of obturation, along with the recognition that no one method of obturation may satisfy all clinical cases. A total of 120 extracted human permanent anterior maxillary and mandibular single rooted teeth were selected for the present study and divided into 3 groups based on the method of obturation technique. Following the preparation the patency at the apical foramen was confirmed by passing a file #15. After obturation of all three groups, teeth were immersed in 1% of aqueous methylene blue dye for a period of two weeks and then samples were subjected to spectrophotometric analysis. The present study was conducted to evaluate in vitro the spectrophotometric analysis to quantitatively analyze relative amount of dye penetration using lateral condensation (Group I), Obtura II (Group II ), Thermafil obturating technique (Group III) with ZOE sealer used in all groups. Teeth obturated with lateral condensation (Group I) shows mean value of 0.0243 and standard deviation of 0.0056. The Group II thermoplasticized injectable moulded Guttapercha (Obtura II) showed 0.0239 mean and standard deviation value of 0.0045 and Group III Thermafil obturation technique shows 0.0189 as mean value and 0.0035 standard deviation values. Following conclusion was drawn from the present study. Group III i.e., Thermafil obturating technique shows minimum mean apical dye penetration compared to Group II (ObturaII) and Group I (lateral condensation).Lateral condensation shows maximum mean apical dye penetration in all three groups.There is no significant difference between the apical dye penetration of lateral condensation and Obtura II. Obturation, lateral condensation, Obtura II, Thermafil, Spectrophotometer, dye penetration. How to cite this article: Samson E, Kulkarni S, Sushil K C, Likhitkar M. An In-Vitro Evaluation and Comparison of Apical Sealing Ability of Three Different Obturation Technique - Lateral Condensation, Obtura II, and Thermafil. J Int Oral Health 2013; 5(2):35-43.

  12. Histopathological Changes in Tissues of Bithynia siamensis goniomphalos Incubated in Crude Extracts of Camellia Seed and Mangosteen Pericarp

    PubMed Central

    Aukkanimart, Ratchadawan; Pinlaor, Somchai; Tesana, Smarn; Aunpromma, Surasit; Booyarat, Chantana; Sriraj, Pranee; Laummaunwai, Porntip; Punjaruk, Wiyada

    2013-01-01

    The present study was performed to observe histopathological changes in tissues of Bithynia siamensis goniomphalos (Gastropoda, Bithyniidae) incubated in crude extract solutions of camellia (Camellia oleifera) seed and mangosteen (Garcinia mangostana) pericarp, and furthermore to estimate the molluscicidal effects of 2 plant substances. Substantial numbers of bithyniid snails were incubated in various concentrations of 2 plant solution for 24 hr. As the positive control, snails incubated in various concentrations of niclosamide, a chemical molluscicide, were used. The histopathological findings were observed in sectioned snail specimens of each experimental and control groups. The results showed that both camellia and mangosteen extracts had molluscicidal effects at 24 hr with 50% lethal concentration (LC50) at concentrations of 0.003 and 0.002 g/ml, respectively, while niclosamide had LC50 at concentrations 0.599 ppm. B. siamensis goniomphalos snail tissues (foot, gill, and digestive system) showed disruption of columnar muscle fibers of the foot, reduction of the length and number of gill cilia, numerous mucous vacuoles, and irregularly shaped of epithelial cells. Irregular apical and calciferous cells, dilatation of the digestive gland tubule, and large hemolymphatic spaces, and irregular apical surfaces, detachment of cilia, and enlargement of lysosomal vacuoles of epidermis were also shown in all groups. By the present study, it is confirmed that 2 plants, camellia and mangosteen, are keeping some substance having molluscicidal effects, and histopathological findings obtained in this study will provide some clues in further studies on their action mechanisms to use them as natural molluscicides. PMID:24327779

  13. Understanding the current state of infection preventionists through competency, role, and activity self-assessment.

    PubMed

    Kalp, Ericka L; Marx, James F; Davis, James

    2017-06-01

    The Association for Professionals in Infection Control and Epidemiology (APIC) MegaSurvey, administered in 2015, was completed by approximately 4,079 APIC members. The survey sought to gain a better understanding the current state of 4 components of infection prevention practice: demographic characteristics, compensation, organizational structure, and practice and competency. The data for this analysis come from the APIC MegaSurvey Practice and Competency domain. Descriptive statistics and χ 2 analyses were conducted to examine differences in infection preventionist (IP) competency, roles, and activity self-assessments. The majority of IPs self-assessed their competency as Proficient compared with Novice or Expert for each of the 8 IP core competency activities. Forty percent of IPs self-rated their competency as Expert in the Preventing/Controlling the Transmission of Infectious Agents/HAIs component. IPs reported Novice competency in Employee/Occupational Health (29%); Cleaning, Sterilization, Disinfection, and Asepsis (23%); and Education and Research categories (22%). Differences in self-rated competency among IPs by discipline type (public health, nurse, and laboratory) were identified. Differences in self-rated competency were identified for each of the 8 IP core competency activities. IPs report using various resource types to gain competency. Future research is needed to identify opportunities to increase competency levels in the weakest-rated competency activities. Copyright © 2017 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

  14. Tolerance of brightness and contrast adjustments on chronic apical abscess and apical granuloma interpretation

    NASA Astrophysics Data System (ADS)

    Purnamasari, L.; Iskandar, H. H. B.; Makes, B. N.

    2017-08-01

    In digitized radiography techniques, adjusting the image enhancement can improve the subjective image quality by optimizing the brightness and contrast for diagnostic needs. To determine the value range of image enhancement (brightness and contrast) on chronic apical abscess and apical granuloma interpretation. 30 periapical radiographs that diagnosed chronic apical abscess and 30 that diagnosed apical granuloma were adjusted by changing brightness and contrast values. The value range of brightness and contrast adjustment that can be tolerated in radiographic interpretations of chronic apical abscess and apical granuloma spans from -10 to +10. Brightness and contrast adjustments on digital radiographs do not affect the radiographic interpretation of chronic apical abscess and apical granuloma if conducted within the value range.

  15. 76 FR 77375 - Airworthiness Directives; Apical Industries, Inc., (Apical) Emergency Float Kits

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-13

    ... Airworthiness Directives; Apical Industries, Inc., (Apical) Emergency Float Kits AGENCY: Federal Aviation... the Apical emergency float kits installed on certain model helicopters under supplemental type... the service information identified in this AD from Apical Industries, Inc., 2608 Temple Heights Drive...

  16. Comparison of the efficacy of nickel-titanium rotary systems with or without the retreatment instruments in the removal of gutta-percha in the apical third

    PubMed Central

    2014-01-01

    Background The purpose of this study was to compare the efficacy of nickel-titanium rotary systems with or without the retreatment instruments in the removal of gutta-percha from the apical third. Methods The systems compared were as follows: ProTaper Universal (PT), ProTaper Universal Retreatment (PTr), Mtwo (M2) and Mtwo Retreatment (M2r). Sixty extracted mandibular incisors were treated with a crown-down technique and filled with gutta-percha and sealer. The apical diameter was standardized in 0.30 mm, 1 mm from the apex. The teeth were distributed into 4 experimental groups: PT, PTr, M2 and M2r. In PTr and M2r groups, filling materials were removed by PTr/M2r followed by root canals preparation up to a PT F4/M2 40; in groups PT/M2, the filling materials were removed and the root canals were prepared by PT up to a PT F4/M2 up to a M2 40. The roots were split and photomicrographing. The percentage of clean area in the apical 5 mm was calculated using software. Data were analyzed with the Kruskal-Wallis test. Results Remaining material was found in all hemisections and there was no statistically significant difference between the groups (p = 0.09). Considering the surface of the canal walls of all teeth, the mean of the percentage of clean area was 54%. Conclusions Considering the applied methodology, remaining filling material was found in all hemisections, regardless of the retreatment technique and PT or M2 were as effective as PTr/PT or M2r/M2. PMID:25128277

  17. Apical root canal transportation of different pathfinding systems and their effects on shaping ability of ProTaper Next

    PubMed Central

    Türker, Sevinç-Aktemur

    2015-01-01

    Background This study aimed to compare glide path preparation of different pathfinding systems and their effects on the apical transportation of ProTaper Next (Dentsply Maillefer, Ballaigues, Switzerland) in mesial root canals of extracted human mandibular molars, using digital subtraction radiography. Material and Methods The mesial canals of 40 mandibular first molars (with curvature angles between 25° and 35°) were selected for this study. The specimens were divided randomly into 4 groups with 10 canals each. Glide paths were created in group 1 with #10, #15 and #20 K-type (Dentsply Maillefer, Ballaigues, Switzerland) stainless steel manual files; in group 2 with Path-File (Dentsply Maillefer) #1, #2, and #3 and in group 3 with #16 ProGlider (Dentsply Maillefer) rotary instruments; in group 4 no glide paths were created. All canals were instrumented up to ProTaper Next X2 to the working length. A double digital radiograph technique was used, pre and post-instrumentation, to assess whether apical transportation and/or aberration in root canal morphology occurred. Instrument failures were also recorded. The data were analyzed statistically using ANOVA and Tukey tests (p<0.05). Results No significant differences were found among groups regarding apical transportation (p>0.05). Two ProTaper Next instruments failed in-group 4. Conclusions Within the parameters of this study, there was no difference between the performance of path-finding files and ProTaper Next system maintained root canal curvature well and was safe to use either with path-finding files or alone. Key words:Glide path, PathFile, ProGlider, ProTaper Next, transportation. PMID:26330936

  18. In vitro comparison of gutta-percha-filled area percentages in root canals instrumented and obturated with different techniques.

    PubMed

    Yilmaz, Ayca; Karagoz-Kucukay, Isil

    2017-01-01

    To evaluate the efficacy of different obturation techniques in root canals instrumented either by hand or rotary instruments with regard to the percentage of gutta- percha-filled area (PGFA). One hundred and sixty extracted mandibular premolars with single, straight root canals were studied. Root canals were prepared to an apical size of 30 by hand with a modified crown-down technique or the ProTaper and HEROShaper systems. Teeth were divided into eight groups (n=20) according to the following instrumentation and obturation techniques: G1: Hand files+lateral condensation (LC), G2: Hand files+Thermafil, G3: ProTaper+LC, G4: ProTaper+single-cone, G5: ProTaper+ProTaper-Obturator, G6: HEROShaper+LC, G7: HEROShaper+single-cone, G8: HEROShaper+HEROfill. Horizontal sections were cut at 1, 3, 5, 7, 9, 11 and 13 mm from the apical foramen. A total of 1120 sections obtained were digitally photographed under a stereomicroscope set at 48X magnification. The cross-sectional area of the canal and the gutta-percha was measured by digital image analysis and the PGFA was calculated for each section. The mean of the PGFA in Thermafil (G2), ProTaper-Obturator (G5) and HEROfill (G8) groups was significantly higher than the other groups. In G3 and G4, PGFA showed no significant difference in the apical segments whereas PGFA was significantly higher at the middle and coronal segments in G3. In G6 and G7, PGFA showed no significant difference in the apical and middle segments whereas PGFA was significantly higher at the coronal segments in G6. The carrier-based gutta-percha obturation systems revealed significantly higher PGFA in comparison to single-cone and lateral condensation techniques.

  19. Apically Extruded Debris in Curved Root Canals Using the WaveOne Gold Reciprocating and Twisted File Adaptive Systems.

    PubMed

    Boijink, Daiana; Costa, Débora Delai; Hoppe, Carolina Bender; Kopper, Patrícia Maria Polli; Grecca, Fabiana Saores

    2018-06-13

    The purpose of this study was to evaluate apical debris extrusion associated with different kinematics in curved root canals. Forty-five extracted mandibular molars with root curvature angles ranging between 20° and 40° and curvature radii <10 mm were randomly assigned to 3 groups (n = 15) according to the kinematics used for root canal preparation: reciprocating motion with the WaveOne Gold system (Dentsply Tulsa Dental, Tulsa, OK), rotary/reciprocating motion with the Twisted File Adaptive system (SybronEndo, Orange, CA), and the manual technique. The debris apically extruded during preparation was collected into preweighed Eppendorf tubes. The tubes were then stored in an incubator at 70°C for 5 days. The extruded debris was quantified by subtracting the preinstrumentation from the postinstrumentation weight of the Eppendorf tubes. The time required for each instrumentation procedure was recorded. Data were analyzed using 1-way analysis of variance Tukey post hoc tests (∝ = 0.05). The WaveOne Gold reciprocating single-file system was associated with less extrusion of debris compared with hand files (P < .05) and the Twisted File Adaptive system (P > .05). The preparation time required by hand files was significantly longer than that required by the other techniques (P < .05). Under the conditions of this study, all of the instrumentation systems caused apical debris extrusion to some degree. The WaveOne Gold reciprocating system was associated with less debris extrusion in curved root canals compared with the manual technique and the Twisted File Adaptive system although the difference between the WaveOne Gold and Twisted File Adaptive systems was not significant. Copyright © 2018 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  20. Internal and external morphology of mandibular molars: An original micro-CT study and meta-analysis with review of implications for endodontic therapy.

    PubMed

    Tomaszewska, Iwona M; Skinningsrud, Bendik; Jarzębska, Anna; Pękala, Jakub R; Tarasiuk, Jacek; Iwanaga, Joe

    2018-03-25

    The aim of this radiological micro-CT study and meta-analysis was to determine the morphological features of the root canal anatomy of the mandibular molars. The radiological study included micro-CT scans of 108 mandibular first, 120 mandibular second, and 146 mandibular third molars. For our meta-analysis, an extensive search was conducted through PubMed, Embase, and Web of Science to identify articles eligible for inclusion. Data extracted included investigative method (cadaveric, intraoperative, or imaging), Vertucci type of canal configuration, presence/number of canals, roots, apical foramina, apical deltas, and intercanal communications. In the mesial roots of mandibular molars, the most frequent Vertucci type of canal configuration was type IV, except for the mandibular third molar where type I was most common. Type I was most common in the distal root. There were usually two canals in the mesial root and one in the distal root. Two was the most common number of roots, and a third root was most prevalent in Asia. One apical foramen was most common in the distal root and two apical foramina in the mesial root. Intercanal communications were most frequent in the mesial root. Knowledge of the complex anatomy of the mandibular molars can make root canal therapy more likely to succeed. We recommend the use of cone-beam computed tomography before and after endodontic treatment to enable the root anatomy to be accurately described and properly diagnosed, and treatment outcome to be assessed. Clin. Anat. 00:000-000, 2018. © 2018 Wiley Periodicals, Inc. © 2018 Wiley Periodicals, Inc.

  1. Micro-Computed Tomography Analysis of the Root Canal Morphology of Palatal Roots of Maxillary First Molars.

    PubMed

    Marceliano-Alves, Marília; Alves, Flávio Rodrigues Ferreira; Mendes, Daniel de Melo; Provenzano, José Claudio

    2016-02-01

    A thorough knowledge of root canal anatomy is critical for successful root canal treatments. This study evaluated the internal anatomy of the palatal roots of maxillary first molars with micro-computed tomography (microCT). The palatal roots of extracted maxillary first molars (n = 169) were scanned with microCT to determine several anatomic parameters, including main canal classification, lateral canal occurrence and location, degree of curvature, main foramen position, apical constriction presence, diameters 1 and 2 mm from the apex and 1 mm from the foramen, minor dentin thickness in those regions, canal volume, surface area, and convexity. All canals were classified as Vertucci type I. The cross sections were oval in 61% of the canals. Lateral canals were found in 25% of the samples. The main foramen did not coincide with the root apex in 95% of the cases. Only 8% of the canals were classified as straight. Apical constriction was identified in 38% of the roots. The minor and major canal diameters and minor dentin thickness were decreased near the apex. The minor dentin thickness 1 mm from the foramen was 0.82 mm. The palatal canals exhibited a volume of 6.91 mm(3) and surface area of 55.31 mm(2) and were rod-shaped. The root canals of the palatal roots were classified as type I. However, some factors need to be considered during the treatment of these roots, including the frequent ocurrence of moderate/severe curvatures, oval-shaped cross-sections, and lateral canals, noncoincidence of the apical foramen with the root apex, and absence of apical constriction in most cases. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  2. Morphogenesis of the mouse neural plate depends on distinct roles of cofilin 1 in apical and basal epithelial domains

    PubMed Central

    Grego-Bessa, Joaquim; Hildebrand, Jeffrey; Anderson, Kathryn V.

    2015-01-01

    The genetic control of mammalian epithelial polarity and dynamics can be studied in vivo at cellular resolution during morphogenesis of the mouse neural tube. The mouse neural plate is a simple epithelium that is transformed into a columnar pseudostratified tube over the course of ∼24 h. Apical F-actin is known to be important for neural tube closure, but the precise roles of actin dynamics in the neural epithelium are not known. To determine how the organization of the neural epithelium and neural tube closure are affected when actin dynamics are blocked, we examined the cellular basis of the neural tube closure defect in mouse mutants that lack the actin-severing protein cofilin 1 (CFL1). Although apical localization of the adherens junctions, the Par complex, the Crumbs complex and SHROOM3 is normal in the mutants, CFL1 has at least two distinct functions in the apical and basal domains of the neural plate. Apically, in the absence of CFL1 myosin light chain does not become phosphorylated, indicating that CFL1 is required for the activation of apical actomyosin required for neural tube closure. On the basal side of the neural plate, loss of CFL1 has the opposite effect on myosin: excess F-actin and myosin accumulate and the ectopic myosin light chain is phosphorylated. The basal accumulation of F-actin is associated with the assembly of ectopic basal tight junctions and focal disruptions of the basement membrane, which eventually lead to a breakdown of epithelial organization. PMID:25742799

  3. A study on FlexMaster : a Ni-Ti rotary engine driven system for root canal preparation.

    PubMed

    Alam, M S; Bashar, A K; Begumr, J A; Kinoshita, J I

    2006-07-01

    In the present study, the effectiveness of recently introduced rotary Nickel Titanium FlexMaster engine driven system was evaluated for cleaning and shaping of root canals in extracted human teeth and compared with that done by the Nickel Titanium Flexofile hand instruments. A total of 40 root canals from 36 extracted mandibular and maxillary teeth with curvature ranging between 20 and 30 degree were divided into two groups, consisting 20 canals in each. FlexMaster instrumentation was done in 20 canals, using crown-down technique and manual filing with Flexofile was done in other 20 canals, using conventional and step-back techniques. 5.25% sodium hypochlorite (NaOCl) solution and 17% ethelene di-amine tetra-acetic acid (EDTA) were used alternatively for irrigation after canal preparation by each instrument size. Time needed for canal preparation of individual canal was recorded. A procedural error such as instrument breakage was also noted. After preparation, all the roots were cut longitudinally by using diamond burs in turbine handpiece and examined under scanning electron microscope. Apical region was quantified for debris and smear layer based on a 5-score numerical evaluation scale. The data established for scoring the debris and smear layer and preparation time of individual canal were analyzed statistically using the Mann-Whitney U test. Completely cleaned root canals were not found with any of the two instruments and none of the instruments maintained the original canal uniformity and regular dentine surface. Because significantly less debris was found in the apical region using the manual filing technique (p< 0.05). Thus the flexmaster system was found to produce less cleaner root canal walls than manual technique. Only one FlexMaster finishing file sized 30 was separated in the apical region during preparation of 30 degree curved canal. The time taken by FlexMaster was significantly better (p< 0.01) than that of hand instruments.

  4. Reproducibility of Echocardiograph-Derived Multilevel Left Ventricular Apical Twist Mechanics.

    PubMed

    Stewart, Glenn M; Yamada, Akira; Kavanagh, Justin J; Haseler, Luke J; Chan, Jonathan; Sabapathy, Surendran

    2016-02-01

    Left ventricular (LV) twist mechanics are routinely assessed via echocardiography in clinical and research trials investigating the function of obliquely oriented myocardial fibers. However, echocardiograph-derived measures of LV twist may be compromised by nonstandardized acquisition of the apical image. This study examined the reproducibility of echocardiograph-derived parameters of apical twist mechanics at multiple levels of the apical myocardium. Two sets of 2D LV parasternal short-axis images were obtained in 30 healthy subjects (24 men; 19-57 year) via echocardiography. Images were acquired immediately distal to the papillary muscles (apical image 1), immediately above the point of LV cavity obliteration at end systole (apical image 3), and midway between apical image 1 and apical image 3 (apical image 2). Repeat scans were performed within 1 hour, and twist mechanics (rotation and rotation rate) were calculated via frame-by-frame tracking of natural acoustic echocardiographic markers (speckle tracking). The magnitude of apical rotation increased progressively toward the apex (apical image 1: 4.2 ± 2.1°, apical image 2: 7.2 ± 3.9°, apical image 3: 11.8 ± 4.6°). apical images 1, 2, and 3 each had moderate to good correlations between repeat scans (ICC: 0.531-0.856). When apical images 1, 2, and 3 were averaged, rotation was 7.7 ± 2.7° and between-scan correlation was excellent (ICC: 0.910). Similar results were observed for systolic and diastolic rotation rates. Averaging multiple standardized apical images, tending progressively toward the apex, generated the most reproducible rotation indices and may be optimal for the assessment of LV twist mechanics across therapeutic, interventional, and research studies; however, care should be taken given the influence of acquisition level on the magnitude of apical rotation. © 2015, Wiley Periodicals, Inc.

  5. Establishing Apical Patency: To be or not to be?

    PubMed

    Mohammadi, Zahed; Jafarzadeh, Hamid; Shalavi, Sousan; Kinoshita, Jun-Ichiro

    2017-04-01

    The apical portion of the root canal is very complex and challenging during endodontic treatment. Root canal preparation and obturation to the apical constriction may provide the best prognosis. Incomplete debridement, foramen transportation, and inadequate seal in the apical portion are considered to be responsible for treatment failure. The technique "apical patency" is considered as a way for maintaining the apical part the free of the debris by recapitulation, using a small K-file through the area of the apical foramen. This term was firstly proposed by Buchanan. In this technique, the smallest diameter file is set 1 mm longer than working length and recapitulated after each instrument to prevent packing of debris in the apical part. Apical patency has been found to be effective in achieving an apical seal with gutta-percha. Teeth prepared with a step back method and with maintained apical patency may show less leakage when obturated with cold lateral condensation technique. Data regarding the effect of apical patency on the healing of periapical tissue are very scarce, and it has been shown that the patency file has detrimental effect on the healing of periapical tissues in animal studies. However, using patency file in endodontic treatment is controversial and further studies are needed. The purpose of this article is to review the effect of using a patency file on the extrusion of root canal contents, the apical seal, postoperative pain, and healing of periapical tissues. Furthermore, the effect of establishing patency on reaching irrigation solutions to the apical portion of the canal and prognosis of root canal treatment are discussed. Keywords: Apical patency, Apical seal, Apical transportation, Postoperative pain, Prognosis.

  6. Inter-dependent apical microtubule and actin dynamics orchestrate centrosome retention and neuronal delamination

    PubMed Central

    Kasioulis, Ioannis

    2017-01-01

    Detachment of newborn neurons from the neuroepithelium is required for correct neuronal architecture and functional circuitry. This process, also known as delamination, involves adherens-junction disassembly and acto-myosin-mediated abscission, during which the centrosome is retained while apical/ciliary membranes are shed. Cell-biological mechanisms mediating delamination are, however, poorly understood. Using live-tissue and super-resolution imaging, we uncover a centrosome-nucleated wheel-like microtubule configuration, aligned with the apical actin cable and adherens-junctions within chick and mouse neuroepithelial cells. These microtubules maintain adherens-junctions while actin maintains microtubules, adherens-junctions and apical end-foot dimensions. During neuronal delamination, acto-myosin constriction generates a tunnel-like actin-microtubule configuration through which the centrosome translocates. This movement requires inter-dependent actin and microtubule activity, and we identify drebrin as a potential coordinator of these cytoskeletal dynamics. Furthermore, centrosome compromise revealed that this organelle is required for delamination. These findings identify new cytoskeletal configurations and regulatory relationships that orchestrate neuronal delamination and may inform mechanisms underlying pathological epithelial cell detachment. PMID:29058679

  7. Drosophila Big bang regulates the apical cytocortex and wing growth through junctional tension.

    PubMed

    Tsoumpekos, Giorgos; Nemetschke, Linda; Knust, Elisabeth

    2018-03-05

    Growth of epithelial tissues is regulated by a plethora of components, including signaling and scaffolding proteins, but also by junctional tension, mediated by the actomyosin cytoskeleton. However, how these players are spatially organized and functionally coordinated is not well understood. Here, we identify the Drosophila melanogaster scaffolding protein Big bang as a novel regulator of growth in epithelial cells of the wing disc by ensuring proper junctional tension. Loss of big bang results in the reduction of the regulatory light chain of nonmuscle myosin, Spaghetti squash. This is associated with an increased apical cell surface, decreased junctional tension, and smaller wings. Strikingly, these phenotypic traits of big bang mutant discs can be rescued by expressing constitutively active Spaghetti squash. Big bang colocalizes with Spaghetti squash in the apical cytocortex and is found in the same protein complex. These results suggest that in epithelial cells of developing wings, the scaffolding protein Big bang controls apical cytocortex organization, which is important for regulating cell shape and tissue growth. © 2018 Tsoumpekos et al.

  8. Cytosolic Na+ Controls an Epithelial Na+ Channel Via the Go Guanine Nucleotide-Binding Regulatory Protein

    NASA Astrophysics Data System (ADS)

    Komwatana, P.; Dinudom, A.; Young, J. A.; Cook, D. I.

    1996-07-01

    In tight Na+-absorbing epithelial cells, the rate of Na+ entry through amiloride-sensitive apical membrane Na+ channels is matched to basolateral Na+ extrusion so that cell Na+ concentration and volume remain steady. Control of this process by regulation of apical Na+ channels has been attributed to changes in cytosolic Ca2+ concentration or pH, secondary to changes in cytosolic Na+ concentration, although cytosolic Cl- seems also to be involved. Using mouse mandibular gland duct cells, we now demonstrate that increasing cytosolic Na+ concentration inhibits apical Na+ channels independent of changes in cytosolic Ca2+, pH, or Cl-, and the effect is blocked by GDP-β -S, pertussis toxin, and antibodies against the α -subunits of guanine nucleotide-binding regulatory proteins (Go). In contrast, the inhibitory effect of cytosolic anions is blocked by antibodies to inhibitory guanine nucleotide-binding regulatory proteins (Gi1/Gi2. It thus appears that apical Na+ channels are regulated by Go and Gi proteins, the activities of which are controlled, respectively, by cytosolic Na+ and Cl-.

  9. Efficacy of Sodium Hypochlorite Activated With Laser in Intracanal Smear Layer Removal: An SEM Study

    PubMed Central

    Shahriari, Shahriar; Kasraei, Shahin; Roshanaei, Ghodratollah; Karkeabadi, Hamed; Davanloo, Hossein

    2017-01-01

    Introduction: The purpose of the present study was to evaluate the different concentrations of sodium hypochlorite activated with laser in removing of the smear layer in the apical, middle, and coronal segments of root canal walls by scanning electron microscopy analysis. Methods: Sixty single-rooted human mandibular teeth were decoronated to a standardized length. The samples were prepared by using Race rotary system to size 40, 0.04 taper and divided into 4 equal groups (n = 15). Group 1, irrigated with EDTA 17% and 5.25% NaOCl, groups 2, 3 and 4, 1%, 2.5%, and 5% NaOCl activated with Nd:YAG laser, respectively. Teeth were split longitudinally and subjected to scanning electron microscope (SEM). Data were analyzed by Kruskal-Wallis, Mann-Whitney tests. P value of <0.05 was considered statistically significant. Results: Five percent NaOCl LAI (laser-activated irrigation) showed best smear layer removal in test groups and the difference was statistically significant (P < 0.001). Control group (EDTA 17% and 5.25% NaOCl irrigation) showed significantly better outcomes in comparative with test groups (P < 0.001). In the apical third, compared to coronal and middle third, the canal walls were often contaminated by inorganic debris and smear layer. Conclusion: All different concentrations of sodium hypochlorite activated with laser have a positive effect on removing of smear layer. Sodium hypochlorite activated with laser removed smear layer more effectively at the coronal and middle third compared to the apical third. PMID:28912942

  10. Effect of gamma-oryzanol on the bioaccessibility and synthesis of cholesterol.

    PubMed

    Mäkynen, K; Chitchumroonchokchai, C; Adisakwattana, S; Failla, M; Ariyapitipun, T

    2012-01-01

    Gamma-oryzanol (gamma-OR) is a unique mixture of triterpene alcohol and sterol ferulates present in rice bran oil. Hypocholesterolemic activity of gamma-OR has been reported in various animal and human studies. However, the mechanisms for this hypocholesterolemic activity of gamma-OR remain unclear. Therefore, the aim of this in vitro study was to examine the effect of gamma-OR on the bioaccessibility and synthesis of cholesterol. The effects of gamma-OR on the efficiency of incorporation of cholesterol into mixed micelles during digestion and apical uptake of cholesterol by Caco-2 human intestinal cells were determined using the coupled in vitro simulated digestion/Caco-2 human intestinal cell model. The impact of gamma-OR on the HMG-CoA reductase activity was also investigated. Although incorporation of cholesterol into synthetic micelles was significantly inhibited by 15-fold molar excess of gamma-OR, efficiency of micellarization of cholesterol during simulated digestion of the rice meal was not significantly altered by the presence of as high as 20-fold molar excess of gamma-OR. Nevertheless, 20-fold molar excess of gamma-OR significantly decreased apical uptake of cholesterol into Caco-2 intestinal cells. In addition, gamma-OR inhibited 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity. These findings suggest that the hypocholesterolemic activity of gamma-OR is due in part to impaired apical uptake of cholesterol into enterocytes and perhaps a decrease in HMG-CoA reductase activity.

  11. [Local immune and oxidative status in exacerbated chronic apical periodontitis].

    PubMed

    Konoplya, A I; Goldobin, D D; Loktionov, A L

    The aim of the study was to define local immune and oxidative changes in patients with exacerbated chronic apical periodontitis. These changes were assessed in saliva of 67 patients with the mean age of 31±2.5 before and after treatment. The study revealed disturbances in cytokines and complement system balance and activation of lipids peroxidation. Combination of Gepon or Vobenzim with Essentiale forte H and Kaskatol proved to be the most effective for correction of this imbalance.

  12. The stochastic nature of action potential backpropagation in apical tuft dendrites.

    PubMed

    Short, Shaina M; Oikonomou, Katerina D; Zhou, Wen-Liang; Acker, Corey D; Popovic, Marko A; Zecevic, Dejan; Antic, Srdjan D

    2017-08-01

    In cortical pyramidal neurons, backpropagating action potentials (bAPs) supply Ca 2+ to synaptic contacts on dendrites. To determine whether the efficacy of AP backpropagation into apical tuft dendrites is stable over time, we performed dendritic Ca 2+ and voltage imaging in rat brain slices. We found that the amplitude of bAP-Ca 2+ in apical tuft branches was unstable, given that it varied from trial to trial (termed "bAP-Ca 2+ flickering"). Small perturbations in dendritic physiology, such as spontaneous synaptic inputs, channel inactivation, or temperature-induced changes in channel kinetics, can cause bAP flickering. In the tuft branches, the density of Na + and K + channels was sufficient to support local initiation of fast spikelets by glutamate iontophoresis. We quantified the time delay between the somatic AP burst and the peak of dendritic Ca 2+ transient in the apical tuft, because this delay is important for induction of spike-timing dependent plasticity. Depending on the frequency of the somatic AP triplets, Ca 2+ signals peaked in the apical tuft 20-50 ms after the 1st AP in the soma. Interestingly, at low frequency (<20 Hz), the Ca 2+ peaked sooner than at high frequency, because only the 1st AP invaded tuft. Activation of dendritic voltage-gated Ca 2+ channels is sensitive to the duration of the dendritic voltage transient. In apical tuft branches, small changes in the duration of bAP voltage waveforms cause disproportionately large increases in dendritic Ca 2+ influx (bAP-Ca 2+ flickering). The stochastic nature of bAP-Ca 2+ adds a new perspective on the mechanisms by which pyramidal neurons combine inputs arriving at different cortical layers. NEW & NOTEWORTHY The bAP-Ca 2+ signal amplitudes in some apical tuft branches randomly vary from moment to moment. In repetitive measurements, successful AP invasions are followed by complete failures. Passive spread of voltage from the apical trunk into the tuft occasionally reaches the threshold for local Na + spike, resulting in stronger Ca 2+ influx. During a burst of three somatic APs, the peak of dendritic Ca 2+ in the apical tuft occurs with a delay of 20-50 ms depending on AP frequency. Copyright © 2017 the American Physiological Society.

  13. In vitro studies on the stability in the proximal gastrointestinal tract and bioaccessibility in Caco-2 cells of chlorogenic acids from spent coffee grounds.

    PubMed

    Monente, Carmen; Ludwig, Iziar A; Stalmach, Angelique; de Peña, Maria Paz; Cid, Concepción; Crozier, Alan

    2015-01-01

    Spent coffee grounds are a potential commercial source of substantial amounts of chlorogenic acids (CGAs). The aim of this study was to evaluate the stability of spent coffee CGAs using in vitro simulated gastroduodenal digestion and to investigate their potential absorption using an in vitro Caco-2 model of human small intestinal epithelium. During in vitro digestion, lactones were partially degraded while caffeoylquinic and feruloylquinic acids were much more stable. Transport and metabolism studies showed that 1% of the total CGAs were absorbed and transported from the apical to the basolateral side of a Caco-2 cell monolayer after 1 h. Lactones and coumaroylquinic acids showed the rate of highest absorption. Caco-2 cells possessed low metabolic activity. In conclusion, spent coffee extracts contain large amounts of CGAs, which remained bioaccessible across the intestinal barrier, albeit to a relatively low degree.

  14. Unusual extraction treatment in Class II division 1 using C-orthodontic mini-implants.

    PubMed

    Chung, Kyu-Rhim; Cho, Jae-Hee; Kim, Seong-Hun; Kook, Yoon-Ah; Cozzani, Mauro

    2007-01-01

    This paper describes the treatment of a female patient, aged 23 years and 5 months, with a Class II division 1 malocclusion, who showed severe anterior protrusion and lower anterior crowding. Specially-designed orthodontic mini-implants were placed bilaterally in the interdental space between both the upper and the lower posterior teeth. Both lower first molars showed severe apical lesions. Therefore, the treatment plan consisted of extraction of both upper first premolars and lower first molars, en masse retraction of the upper six anterior teeth, lower anterior alignment, and protraction of all the lower molars. C-implants(R) were used as substitutes for maxillary posterior anchorage teeth during anterior retraction and as hooks for mandibular molar protraction. The correct overbite and overjet were obtained by intruding and retracting the upper six anterior teeth into their proper positions. The dentition was detailed using conventional orthodontic appliances. The upper C-implants contributed to an improvement in facial balance, and the lower C-implants made it possible to protract the lower second and third molars with less effect on the axis of the lower anterior teeth. The active treatment period was 29 months and the patient's teeth continued to be stable 11 months after debonding.

  15. Physiological integration enhanced the tolerance of Cynodon dactylon to flooding.

    PubMed

    Li, Z J; Fan, D Y; Chen, F Q; Yuan, Q Y; Chow, W S; Xie, Z Q

    2015-03-01

    Many flooding-tolerant species are clonal plants; however, the effects of physiological integration on plant responses to flooding have received limited attention. We hypothesise that flooding can trigger changes in metabolism of carbohydrates and ROS (reactive oxygen species) in clonal plants, and that physiological integration can ameliorate the adverse effects of stress, subsequently restoring the growth of flooded ramets. In the present study, we conducted a factorial experiment combining flooding to apical ramets and stolon severing (preventing physiological integration) between apical and basal ramets of Cynodon dactylon, which is a stoloniferous perennial grass with considerable flooding tolerance. Flooding-induced responses including decreased root biomass, accumulation of soluble sugar and starch, as well as increased activity of superoxide dismutase (SOD) and ascorbate peroxidase (APX) in apical ramets. Physiological integration relieved growth inhibition, carbohydrate accumulation and induction of antioxidant enzyme activity in stressed ramets, as expected, without any observable cost in unstressed ramets. We speculate that relief of flooding stress in clonal plants may rely on oxidising power and electron acceptors transferred between ramets through physiological integration. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

  16. Prediction of intestinal absorption and metabolism of pharmacologically active flavones and flavanones.

    PubMed

    Serra, H; Mendes, T; Bronze, M R; Simplício, Ana Luísa

    2008-04-01

    Three glycosilated flavonoids (diosmin, hesperidin and naringin) and respective aglycones were characterized in terms of their apparent ionisation constants and bidirectional permeability using the cellular model Caco-2 as well as the artificial membrane model PAMPA. Ionisation curves were established by capillary electrophoresis. It was confirmed that significant amounts of the aglycones are ionised at physiological pH whereas the glycosides are in the neutral form. Permeation was not detected for the glycosides in either the apical-to-basolateral or basolateral-to-apical directions confirming the need for metabolism before absorption through the intestinal membrane. The aglycones permeated in both directions with apparent permeabilities (P(app)) in the range of 1-8x10(-5) cm/s. The results from both in vitro methods correlated providing some evidence of passive transport; however, the hypothesis of active transport cannot be excluded particularly in the case of diosmetin. Metabolism of the aglycones was detected with the cell model, more extensively when loading in the apical side. Some of the metabolites were identified as glucuronide conjugates by enzymatic hydrolysis.

  17. Chromatin remodeling in stem cell maintenance in Arabidopsis thaliana.

    PubMed

    Shen, Wen-Hui; Xu, Lin

    2009-07-01

    Pluripotent stem cells are able to both self-renew and generate undifferentiated cells for the formation of new tissues and organs. In higher plants, stem cells found in the shoot apical meristem (SAM) and the root apical meristem (RAM) are origins of organogenesis occurring post-embryonically. It is important to understand how the regulation of stem cell fate is coordinated to enable the meristem to constantly generate different types of lateral organs. Much knowledge has accumulated on specific transcription factors controlling SAM and RAM activity. Here, we review recent evidences for a role of chromatin remodeling in the maintenance of stable expression states of transcription factor genes and the control of stem cell activity in Arabidopsis.

  18. Sweet taste receptors in rat small intestine stimulate glucose absorption through apical GLUT2.

    PubMed

    Mace, Oliver J; Affleck, Julie; Patel, Nick; Kellett, George L

    2007-07-01

    Natural sugars and artificial sweeteners are sensed by receptors in taste buds. T2R bitter and T1R sweet taste receptors are coupled through G-proteins, alpha-gustducin and transducin, to activate phospholipase C beta2 and increase intracellular calcium concentration. Intestinal brush cells or solitary chemosensory cells (SCCs) have a structure similar to lingual taste cells and strongly express alpha-gustducin. It has therefore been suggested over the last decade that brush cells may participate in sugar sensing by a mechanism analogous to that in taste buds. We provide here functional evidence for an intestinal sensing system based on lingual taste receptors. Western blotting and immunocytochemistry revealed that all T1R members are expressed in rat jejunum at strategic locations including Paneth cells, SCCs or the apical membrane of enterocytes; T1Rs are colocalized with each other and with alpha-gustducin, transducin or phospholipase C beta2 to different extents. Intestinal glucose absorption consists of two components: one is classical active Na+-glucose cotransport, the other is the diffusive apical GLUT2 pathway. Artificial sweeteners increase glucose absorption in the order acesulfame potassium approximately sucralose > saccharin, in parallel with their ability to increase intracellular calcium concentration. Stimulation occurs within minutes by an increase in apical GLUT2, which correlates with reciprocal regulation of T1R2, T1R3 and alpha-gustducin versus T1R1, transducin and phospholipase C beta2. Our observation that artificial sweeteners are nutritionally active, because they can signal to a functional taste reception system to increase sugar absorption during a meal, has wide implications for nutrient sensing and nutrition in the treatment of obesity and diabetes.

  19. Co-localization of endogenous Arf6 and its activator EFA6D in the granular convoluted tubule cells of mouse submandibular glands under normal conditions and when stimulated by isoproterenol, noradrenaline and carbachol.

    PubMed

    Tachow, Apussara; Thoungseabyoun, Wipawee; Phuapittayalert, Laorrat; Petcharat, Kanoktip; Sakagami, Hiroyuki; Kondo, Hisatake; Hipkaeo, Wiphawi

    2017-10-01

    This study proposed to investigate the localization at light and electron microscopic levels of Arf6 and its activator EFA6D in the mouse submandibular gland (SMG) under normal conditions and when stimulated by adrenergic or cholinergic agonists. SMGs of male adult mice were utilized for immunoblotting and immuno-light and -electron microscopic analyses. Isoproterenol and noradrenalin were used as adrenergics, while carbachol was used for the cholinergic stimulant. SMGs were examined at 15, 30, 60 and 120min after intraperitoneal injection of these agents. Immunoreactivities for both Arf6 and its activator EFA6D were similarly intense in the basolateral domain of GCTs, but no significant immunoreactivities were seen in the apical domain of GCT cells or any domain of acinar cells under normal conditions. In immuno-electron microscopy, the immunoreactive materials were mainly deposited on the basolateral plasma membranes and subjacent cytoplasm. Shortly after injection of isoproterenol and noradrenaline, but not carbachol, the immunoreactivities for both molecules were additionally seen on the apical plasmalemma of most, if not all, GCT cells, but not acinar cells. The present findings suggest that the direct involvement of Arf6/EFA6D in regulatory exocytosis at the apical plasma membrane of acinar and GCT cells is apparently to be smaller, if present, than that of endocytosis at the basolateral membranes of GCT cells under normal conditions. This also suggests that the two molecules function additionally at the apical membrane of GCT cells for modulation of saliva secretion under β-adrenoceptor stimulation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Comparative evaluation of apically extruded debris during root canal instrumentation using two Ni-Ti single file rotary systems: An in vitro study

    PubMed Central

    Singbal, Kiran; Jain, Disha; Raja, Kranthi; Hoe, Tan Ming

    2017-01-01

    Background: Apical extrusion of debris during instrumentation is detrimental to the patient. Aim: The aim of this study was to evaluate the apical extrusion of debris during root canal instrumentation using two single file rotary Ni-Ti systems. Materials and Methods: Thirty freshly extracted mandibular premolars with straight roots were sterilized and divided into two groups instrumented using: One Shape rotary Ni-Ti system with Endoflare orifice shaper (Group 1) and Neo-Niti rotary Ni-Ti system with C1 orifice shaper (Group 2). Preweighed Eppendorf tubes fitted for each tooth before instrumentation. During instrumentation, 1 mL of distilled water with a 30-gauge needle was used to irrigate after every instrument. Tips of the tooth were irrigated with 2 ml distilled water after removal from Eppendorf tubes. The total volume of irrigant in each group was the same 8 ml. All tubes were incubated at 68°C for 15 days and subsequently weighed. The difference between pre- and post-debris weights was calculated, and statistical analysis was performed using independent t-test and level of significance was set at 0.05. Results: The difference between pre- and post-weights was significantly greater for the One Shape system. Conclusions: The Neolix Niti single file was associated with less extrusion compared to One Shape single file system. PMID:28855748

  1. A simple model to demonstrate the electronic apex locator.

    PubMed

    Tinaz, A C; Alaçam, T; Topuz, O

    2002-11-01

    To describe and evaluate a newly developed model for demonstrating and teaching the use of electronic apex locators. A phantom model, master jaw model and extracted human teeth were used to construct the demonstration model with alginate impression material as the periapical conductive medium. The model was validated in a series of length determinations with apical foramina enlarged to 0.20, 0.30 and 0.45 mm diameter, and the stability of the model was evaluated up to 45 h after construction. All evaluations were conducted with the Root ZX apex locator with 2.65 and 5.25% NaOCl in the canals. Most length measurements were within 1 mm of actual root length (range: -2.2 to +0.21 mm) and did not change significantly over 45 h for teeth with foramina of 0.3 mm or less. Measurements for teeth with wide (0.45 mm) apices were stable up to 28 h. NaOCl concentration did not significantly affect the readings. A simple, inexpensive model can be manufactured from plastic dental jaws, natural teeth and alginate impression material to demonstrate electronic working length measurement. The model is stable for many hours and provides consistent results with different concentrations of NaOCl in the canal and various apical diameters. The model is a useful teaching aid but needs further evaluation and refinement before use in research applications.

  2. Comparative evaluation of the amount of debris extruded apically using conventional syringe, passive ultrasonic irrigation and EndoIrrigator Plus system: An in vitro study

    PubMed Central

    Shetty, Vidhi Prabhakar; Naik, Balaram Damodar; Pachlag, Amit Kashinath; Yeli, Mahantesh Mrityunjay

    2017-01-01

    Aim: The aim of this study is to compare the effects of conventional syringe, passive ultrasonic irrigation (PUI), and EndoIrrigator Plus on the amount of apically extruded debris. Materials and Methods: Thirty extracted human mandibular premolars were selected and randomly assigned to three groups (n = 10). The root canals were irrigated with conventional syringe, PUI, and EndoIrrigator Plus. Sodium hypochlorite was used as an irrigant, and debris was collected in a previously described experimental model (Myers and Montgomery 1991). It was then stored in an incubator at 37°C for 10 days to evaporate the irrigant before weighing the dry debris. The mean weight of debris was assessed, one-way analysis of variance was used for comparison of values, and post hoc Tukey's test was used between groups (P = 0.05). Results: The EndoIrrigator Plus group extruded significantly less debris than PUI and conventional syringe groups (P < 0.05). Furthermore, PUI group extruded significantly less debris than conventional syringe irrigation group (P < 0.05). Conclusions: 1. All the three irrigation systems were associated with apical extrusion of debris, 2. EndoIrrigator Plus system extruded significantly less debris than the PUI system and the conventional syringe irrigation system, 3. PUI system extruded significantly less debris than the conventional syringe irrigation system. PMID:29430092

  3. Comparative evaluation of apically extruded debris during root canal instrumentation using two Ni-Ti single file rotary systems: An in vitro study.

    PubMed

    Singbal, Kiran; Jain, Disha; Raja, Kranthi; Hoe, Tan Ming

    2017-01-01

    Apical extrusion of debris during instrumentation is detrimental to the patient. The aim of this study was to evaluate the apical extrusion of debris during root canal instrumentation using two single file rotary Ni-Ti systems. Thirty freshly extracted mandibular premolars with straight roots were sterilized and divided into two groups instrumented using: One Shape rotary Ni-Ti system with Endoflare orifice shaper (Group 1) and Neo-Niti rotary Ni-Ti system with C1 orifice shaper (Group 2). Preweighed Eppendorf tubes fitted for each tooth before instrumentation. During instrumentation, 1 mL of distilled water with a 30-gauge needle was used to irrigate after every instrument. Tips of the tooth were irrigated with 2 ml distilled water after removal from Eppendorf tubes. The total volume of irrigant in each group was the same 8 ml. All tubes were incubated at 68°C for 15 days and subsequently weighed. The difference between pre- and post-debris weights was calculated, and statistical analysis was performed using independent t -test and level of significance was set at 0.05. The difference between pre- and post-weights was significantly greater for the One Shape system. The Neolix Niti single file was associated with less extrusion compared to One Shape single file system.

  4. Comparison of two methods of digital imaging technology for small diameter K-file length determination.

    PubMed

    Maryam, Ehsani; Farida, Abesi; Farhad, Akbarzade; Soraya, Khafri

    2013-11-01

    Obtaining the proper working length in endodontic treatment is essential. The aim of this study was to compare the working length (WL) assessment of small diameter K-files using the two different digital imaging methods. The samples for this in-vitro experimental study consisted of 40 extracted single-rooted premolars. After access cavity preparation, the ISO files no. 6, 8, and 10 stainless steel K-files were inserted in the canals in the three different lengths to evaluate the results in a blinded manner: At the level of apical foramen(actual)1 mm short of apical foramen2 mm short of apical foramen A digital caliper was used to measure the length of the files which was considered as the Gold Standard. Five observers (two oral and maxillofacial radiologists and three endodontists) observed the digital radiographs which were obtained using PSP and CCD digital imaging sensors. The collected data were analyzed by SPSS 17 and Repeated Measures Paired T-test. In WL assessment of small diameter K-files, a significant statistical relationship was seen among the observers of two digital imaging techniques (P<0.001). However, no significant difference was observed between the two digital techniques in WL assessment of small diameter K-files (P<0.05). PSP and CCD digital imaging techniques were similar in WL assessment of canals using no. 6, 8, and 10 K-files.

  5. Functional, morphological and electrocardiographical abnormalities in patients with apical hypertrophic cardiomyopathy and apical aneurysm: correlation with cardiac MR

    PubMed Central

    Suwa, Kenichiro; Satoh, Hiroshi; Sano, Makoto; Nobuhara, Mamoru; Saitoh, Takeji; Saotome, Masao; Urushida, Tsuyoshi; Katoh, Hideki; Tawarahara, Kei; Ohtani, Hayato; Wakabayashi, Yasushi; Takase, Hiroyuki; Terada, Hajime; Takehara, Yasuo; Sakahara, Harumi; Hayashi, Hideharu

    2014-01-01

    Objective The prognosis of apical hypertrophic cardiomyopathy (APH) has been benign, but apical myocardial injury has prognostic importance. We studied functional, morphological and electrocardiographical abnormalities in patients with APH and with apical aneurysm and sought to find parameters that relate to apical myocardial injury. Methods Study design: a multicentre trans-sectional study. Patients: 45 patients with APH and 5 with apical aneurysm diagnosed with transthoracic echocardiography (TTE) in the database of Hamamatsu Circulation Forum. Measure: the apical contraction with cine-cardiac MR (CMR), the myocardial fibrotic scar with late gadolinium enhancement (LGE)-CMR, and QRS fragmentation (fQRS) defined when two ECG-leads exhibited RSR’s patterns. Results Cine-CMR revealed 27 patients with normal, 12 with hypokinetic and 11 with dyskinetic apical contraction. TTE misdiagnosed 11 (48%) patients with hypokinetic and dyskinetic contraction as those with normal contraction. Apical LGE was apparent in 10 (83%) and 11 (100%) patients with hypokinetic and dyskinetic contraction, whereas only in 11 patients (41%) with normal contraction (p<0.01). Patients with dyskinetic apical contraction had the lowest left ventricular ejection fraction, the highest prevalence of ventricular tachycardia, and the smallest ST depression and depth of negative T waves. The presence of fQRS was associated with impaired apical contraction and apical LGE (OR=8.32 and 8.61, p<0.05). Conclusions CMR is superior to TTE for analysing abnormalities of the apex in patients with APH and with apical aneurysm. The presence of fQRS can be a promising parameter for the early detection of apical myocardial injury. PMID:25332823

  6. Biological effects of compressive forces exerted on particulate bone grafts during socket preservation: animal study.

    PubMed

    Delgado-Ruiz, Rafael; Romanos, Georgios E; Alexandre Gerhke, Sergio; Gomez-Moreno, Gerardo; Maté-Sánchez de Val, José Eduardo; Calvo-Guirado, José Luis

    2016-08-02

    To compare different compressive forces exerted on a particulate graft material during socket preservation and their effects on bone regeneration. Six male dogs were used. The second, third, and fourth premolars, and the first molar were extracted bilaterally at the lower jaws. A particulate synthetic biphasic grafting material (60% HA and 40% β-tricalcium phosphate) was used. Three different standardized compressive forces were applied randomly during the socket preservation. The sample was divided into four experimental groups Test A (10 g), Test B (50 g), Test C (200 g), and Control (empty sockets). Collagen membranes were placed, and primary closure was obtained. Two months after the surgery the animals were sacrificed, and histomorphometric analysis of non-decalcified samples was performed at the coronal, middle, and apical thirds. Grafted sockets resulted in higher bony contour (3 ± 0.43 mm 2 ; P < 0.05). The particles penetrated up to the apical third in the group C but not in the other test groups and controls (P < 0.05). The percentage of new bone were higher at the coronal and apical thirds for Controls and group C compared to A and B groups (P < 0.05). The residual graft was higher for group C (53 ± 1.4%), followed by group B (45 ± 3.1%) and group A (35 ± 1.9%; P < 0.05). The percentages of connective tissue were higher at the middle third without differences between groups (P > 0.05). Within the limitations of this experimental animal study, it might be concluded that grafted sockets compressed with 200 g force will have higher bony contours; higher compressive forces facilitate the penetration of the particulate graft material into the apical area of the socket and results in more bone formation at the coronal, middle, and apical thirds. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  7. Efficacy of Different Methods for Removing Root Canal Filling Material in Retreatment - An In-vitro Study.

    PubMed

    Kasam, Swetha; Mariswamy, Annapoorna Ballagere

    2016-06-01

    Although success of endodontic therapy has significantly improved in the last few decades due to the introduction of novel materials and techniques, failures of endodontic therapy requiring re-treatment still comprise a significant percentage of patients requiring root canal treatment. To evaluate and compare the effective removal of gutta percha and sealer, amount of apical debris extrusion and time required for gutta percha removal using various endodontic files. Total 48 extracted mandibular premolars were mounted on acrylic blocks and endodontic procedure was carried out using size 40 K file and obturated using guttapercha and zinc oxide eugenol sealer. After one month storage, samples were decoronated, mounted on screw capped vials and subjected to removal of obturated material by four instruments: H files, safe sided H files, protaper universal retreatment rotary system and ultrasonic retreatment tip, grouped as 1, 2, 3, and 4 respectively. Only 2mm of obturated material from the coronal part was removed using no. 3 Gates Glidden drill, guttapercha was softened with a drop of xylene for 2 mins for each canal and retreatment was performed. The retreatment procedure was said to be complete when no visible debris were observed on the instrument flutes. The samples split into two halves and examined under stereomicroscope, photographed, assessed using AUTOCAD software and percentage of remaining filling material in coronal, middle, apical thirds of the canal was calculated in mm(2). Retreatment time was recorded in seconds and apically extruded debris was assessed by microbalance in grams for each tooth. The data was analyzed by using descriptive statistics, ANOVA and Scheffe's post hoc test through SPSS for windows (v 16.0). The ultrasonic retreatment tip had less percentage of residual guttapercha/sealer, shorter mean operating time and little apical extrusion with a significant difference (p<0.05) between the other groups. All techniques retained guttapercha/sealer remnants within the root canal. The ultrasonic retreatment tip proved to be an efficient method of removing obturated material. It was fastest with least apical debris extrusion.

  8. Efficacy of Different Methods for Removing Root Canal Filling Material in Retreatment - An In-vitro Study

    PubMed Central

    Mariswamy, Annapoorna Ballagere

    2016-01-01

    Introduction Although success of endodontic therapy has significantly improved in the last few decades due to the introduction of novel materials and techniques, failures of endodontic therapy requiring re-treatment still comprise a significant percentage of patients requiring root canal treatment. Aim To evaluate and compare the effective removal of gutta percha and sealer, amount of apical debris extrusion and time required for gutta percha removal using various endodontic files. Materials and Methods Total 48 extracted mandibular premolars were mounted on acrylic blocks and endodontic procedure was carried out using size 40 K file and obturated using guttapercha and zinc oxide eugenol sealer. After one month storage, samples were decoronated, mounted on screw capped vials and subjected to removal of obturated material by four instruments: H files, safe sided H files, protaper universal retreatment rotary system and ultrasonic retreatment tip, grouped as 1, 2, 3, and 4 respectively. Only 2mm of obturated material from the coronal part was removed using no. 3 Gates Glidden drill, guttapercha was softened with a drop of xylene for 2 mins for each canal and retreatment was performed. The retreatment procedure was said to be complete when no visible debris were observed on the instrument flutes. The samples split into two halves and examined under stereomicroscope, photographed, assessed using AUTOCAD software and percentage of remaining filling material in coronal, middle, apical thirds of the canal was calculated in mm2. Retreatment time was recorded in seconds and apically extruded debris was assessed by microbalance in grams for each tooth. The data was analyzed by using descriptive statistics, ANOVA and Scheffe’s post hoc test through SPSS for windows (v 16.0). Results The ultrasonic retreatment tip had less percentage of residual guttapercha/sealer, shorter mean operating time and little apical extrusion with a significant difference (p<0.05) between the other groups. Conclusion All techniques retained guttapercha/sealer remnants within the root canal. The ultrasonic retreatment tip proved to be an efficient method of removing obturated material. It was fastest with least apical debris extrusion. PMID:27504397

  9. A Comparative SEM Investigation of Smear Layer Remaining on Dentinal Walls by Three Rotary NiTi Files with Different Cross Sectional Designs in Moderately Curved Canals

    PubMed Central

    Kakkar, Pooja; Vats, Asit

    2015-01-01

    Objective: The objective of this study was to compare the smear layer formed on root canal walls during canal preparation of extracted human teeth by Twisted, Mtwo, and ProTaper rotary nickel titanium instruments. Materials and Methods: Sixty single rooted human premolar teeth with root curvature <250 were selected and randomly divided into three Groups (n= 20 teeth per Group). Three types of rotary nickel titanium instruments were used, Twisted (SybronEndo, Orange, CA, USA), Mtwo (VDW, Munich, Germany) and ProTaper (Dentsply Maillefer, Ballaigues, Switzerland) according to manufacturer’s instructions to instrument the root canals. Irrigation for all groups was performed after each instrument change with 3ml of 3% sodium hypochlorite followed by Glyde (File Prep, Dentsply, Maillefer, Ballaigues, Switzerland) as chelator paste and lubricant. Three different areas (coronal, middle and apical thirds) of the root canal were evaluated using scanning electron microscopy (SEM). The canal wall of each sample was assessed and compared using a predefined scale for the presence or absence of smear layer. Data were analysed statistically using ANOVA and Tukey HSD test Results: All three groups showed statistically significant more smear layer in the apical thirds of the canal as compared to the coronal and middle thirds (p<0.001). Mtwo rotary file system produced significantly less smear layer (p<0.001) compared to Twisted and ProTaper rotary instruments in the apical portion. Twisted Files resulted in less smear layer formation in the apical thirds of the canal compared to ProTaper rotary instruments but were statistically insignificant. Conclusion: Completely clean root canals were not found after instrumentation with any of the three instruments. Under the confines of this study Mtwo instruments produced significantly cleaner dentin wall surfaces throughout the canal length in comparison to Twisted and ProTaper rotary files. Twisted Files proved to be comparable to ProTaper rotary instruments with respect to canal cleanliness in the apical thirds of the root canal. PMID:25954704

  10. The role of gravity in apical dominance: effects of clinostating on shoot inversion-induced ethylene production, shoot elongation and lateral bud growth

    NASA Technical Reports Server (NTRS)

    Prasad, T. K.; Cline, M. G.

    1987-01-01

    Shoot inversion-induced release of apical dominance in Pharbitis nil is inhibited by rotating the plant at 0.42 revolutions per minute in a vertical plane perpendicular to the axis of rotation of a horizontal clinostat. Clinostating prevented lateral bud outgrowth, apparently by negating the restriction of the shoot elongation via reduction of ethylene production in the inverted shoot. Radial stem expansion was also decreased. Data from experiments with intact tissue and isolated segments indicated that shoot-inversion stimulates ethylene production by increasing the activity of 1-aminocyclopropane-1-carboxylic acid synthase. The results support the hypothesis that shoot inversion-induced release of apical dominance in Pharbitis nil is due to gravity stress and is mediated by ethylene-induced retardation of the elongation of the inverted shoot.

  11. A Novel Approach for Apical Dissection During Robot-assisted Radical Prostatectomy: The "Collar" Technique.

    PubMed

    Bianchi, Lorenzo; Turri, Filippo Maria; Larcher, Alessandro; De Groote, Ruben; De Bruyne, Peter; De Coninck, Vincent; Goossens, Marijn; D'Hondt, Frederiek; De Naeyer, Geert; Schatteman, Peter; Mottrie, Alexandre

    2018-02-02

    Apical dissection in robot-assisted radical prostatectomy (RARP) affects not only cancer control, but also continence recovery. To describe a novel approach for apical dissection, the collar technique, to reduce apical positive surgical margins (PSMs). A total of 189 consecutive patients (81 in the control group, 108 in the collar technique group) underwent RARP at a single center. rates of apical PSMs; secondary outcome: urinary continence. The urethral sphincter complex is incised 2-3mm distally to the apex, to stay farther from it and reduce PSMs; the underlying smooth muscle is exposed and incised closer to the apex to preserve the maximal length of the lissosphincter. Mann-Whitney U and chi-square tests compared median and proportions between the two groups, respectively. Univariate logistic regression tested the association between technique employed and risk of apical PSMs. Fourteen patients (7.4%) revealed apical PSMs (9.9% in the control group, 5.6% in the collar group; p=0.7). When the collar technique was used, significantly lower rates of apical PSMs occurred in pT2 disease (0% vs 7.1%; p=0.03). In case of apical tumor at preoperative magnetic resonance imaging (MRI; n=43), the collar technique determined significantly lower overall (9.7% vs 42%) and apical (3.2% vs 42%) PSMs (all p≤0.02). Continence recovery in the collar and control groups was similar. When preoperative MRI showed an apical tumor, the collar technique had a significantly lower risk of apical PSMs (odds ratio: 0.05, p=0.009). The collar technique reduces the rates of apical PSMs in case of apical tumor, preserving the length of the lissosphincter. We describe a novel approach for apical dissection during robot-assisted radical prostatectomy. Our technique reduces the rates of apical surgical margins in case of apical tumor at preoperative magnetic resonance imaging and leads to optimal continence recovery. Copyright © 2018 European Association of Urology. Published by Elsevier B.V. All rights reserved.

  12. Barrier role of actin filaments in regulated mucin secretion from airway goblet cells.

    PubMed

    Ehre, Camille; Rossi, Andrea H; Abdullah, Lubna H; De Pestel, Kathleen; Hill, Sandra; Olsen, John C; Davis, C William

    2005-01-01

    Airway goblet cells secrete mucin onto mucosal surfaces under the regulation of an apical, phospholipase C/G(q)-coupled P2Y(2) receptor. We tested whether cortical actin filaments negatively regulate exocytosis in goblet cells by forming a barrier between secretory granules and plasma membrane docking sites as postulated for other secretory cells. Immunostaining of human lung tissues and SPOC1 cells (an epithelial, mucin-secreting cell line) revealed an apical distribution of beta- and gamma-actin in ciliated and goblet cells. In goblet cells, actin appeared as a prominent subplasmalemmal sheet lying between granules and the apical membrane, and it disappeared from SPOC1 cells activated by purinergic agonist. Disruption of actin filaments with latrunculin A stimulated SPOC1 cell mucin secretion under basal and agonist-activated conditions, whereas stabilization with jasplakinolide or overexpression of beta- or gamma-actin conjugated to yellow fluorescent protein (YFP) inhibited secretion. Myristoylated alanine-rich C kinase substrate, a PKC-activated actin-plasma membrane tethering protein, was phosphorylated after agonist stimulation, suggesting a translocation to the cytosol. Scinderin (or adseverin), a Ca(2+)-activated actin filament severing and capping protein was cloned from human airway and SPOC1 cells, and synthetic peptides corresponding to its actin-binding domains inhibited mucin secretion. We conclude that actin filaments negatively regulate mucin secretion basally in airway goblet cells and are dynamically remodeled in agonist-stimulated cells to promote exocytosis.

  13. Ineffective and prolonged apical contraction is associated with chest pain and ischaemia in apical hypertrophic cardiomyopathy.

    PubMed

    Stephenson, Edward; Monney, Pierre; Pugliese, Francesca; Malcolmson, James; Petersen, Steffen E; Knight, Charles; Mills, Peter; Wragg, Andrew; O'Mahony, Constantinos; Sekhri, Neha; Mohiddin, Saidi A

    2018-01-15

    To investigate the hypothesis that persistence of apical contraction into diastole is linked to reduced myocardial perfusion and chest pain. Apical hypertrophic cardiomyopathy (HCM) is defined by left ventricular (LV) hypertrophy predominantly of the apex. Hyperdynamic contractility resulting in obliteration of the apical cavity is often present. Apical HCM can lead to drug-refractory chest pain. We retrospectively studied 126 subjects; 76 with apical HCM and 50 controls (31 with asymmetrical septal hypertrophy (ASH) and 19 with non-cardiac chest pain and culprit free angiograms and structurally normal hearts). Perfusion cardiac magnetic resonance imaging (CMR) scans were assessed for myocardial perfusion reserve index (MPRi), late gadolinium enhancement (LGE), LV volumes (muscle and cavity) and regional contractile persistence (apex, mid and basal LV). In apical HCM, apical MPRi was lower than in normal and ASH controls (p<0.05). In apical HCM, duration of contractile persistence was associated with lower MPRi (p<0.01) and chest pain (p<0.05). In multivariate regression, contractile persistence was independently associated with chest pain (p<0.01) and reduced MPRi (p<0.001). In apical HCM, regional contractile persistence is associated with impaired myocardial perfusion and chest pain. As apical myocardium makes limited contributions to stroke volume, apical contractility is also largely ineffective. Interventions to reduce apical contraction and/or muscle mass are potential therapies for improving symptoms without reducing cardiac output. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Apical P2XR contribute to [Ca2+]i signaling and Isc in mouse renal MCD.

    PubMed

    Li, Liuzhe; Lynch, I Jeanette; Zheng, Wencui; Cash, Melanie N; Teng, Xueling; Wingo, Charles S; Verlander, Jill W; Xia, Shen-Ling

    2007-08-03

    We examined P2X receptor expression and distribution in the mouse collecting duct (CD) and their functional role in Ca(2+) signaling. Both P2X(1) and P2X(4) were detected by RT-PCR and Western blot. Immunohistochemistry demonstrated apical P2X(1) and P2X(4) immunoreactivity in principal cells in the outer medullary CD (OMCD) and inner medullary CD (IMCD). Luminal ATP induced an increase in Ca(2+) signaling in native medullary CD (MCD) as measured by fluorescence imaging. ATP also induced an increase in Ca(2+) signaling in MCD cells grown in primary culture but not in the presence of P2XR antagonist PPNDS. Short circuit current (I(sc)) measurement with mouse IMCD cells showed that P2XR agonist BzATP induced a larger I(sc) than did P2YR agonist UTP in the apical membrane. Our data reveal for the first time that P2X(1) and P2X(4) are cell-specific with prominent immunoreactivity in the apical area of MCD cells. The finding that P2XR blockade inhibits ATP-induced Ca(2+) signaling suggests that activation of P2XR is a key step in Ca(2+)-dependent purinergic signaling. The result that activation of P2XR produces large I(sc) indicates the necessity of P2XR in renal CD ion transport.

  15. Fabry disease presenting as apical left ventricular hypertrophy in a patient carrying the missense mutation R118C.

    PubMed

    Caetano, Francisca; Botelho, Ana; Mota, Paula; Silva, Joana; Leitão Marques, António

    2014-03-01

    Anderson-Fabry disease is an X-linked lysosomal storage disorder caused by abnormalities of the GLA gene, which encodes the enzyme α-galactosidase A. A deficiency of this enzyme leads to the lysosomal accumulation of glycosphingolipids, which may cause left ventricular hypertrophy that is typically concentric and symmetric. We present the case of a 60-year-old woman with symptoms of dyspnea, atypical chest pain and palpitations, in whom a transthoracic echocardiogram revealed an apical variant of hypertrophic cardiomyopathy. Analysis of specific sarcomeric genetic mutations was negative. The patient underwent a screening protocol for Anderson-Fabry disease, using a dried blood spot test, which was standard at our institution for patients with left ventricular hypertrophy. The enzymatic activity assay revealed reduced α-galactosidase A enzymatic activity. Molecular analysis identified a missense point mutation in the GLA gene (p.R118C). This case report shows that Anderson-Fabry disease may cause an apical form of left ventricular hypertrophy. The diagnosis was only achieved because of systematic screening, which highlights the importance of screening for Anderson-Fabry disease in patients with unexplained left ventricular hypertrophy, including those presenting with more unusual patterns, such as apical variants of left ventricular hypertrophy. This case also supports the idea that the missense mutation R118C is indeed a true pathogenic mutation of Anderson-Fabry disease. Copyright © 2012 Sociedade Portuguesa de Cardiologia. Published by Elsevier España. All rights reserved.

  16. Trichomes and chemical composition of the volatile oil of Trichogonia cinerea (Gardner) R. M. King & H. Rob. (Eupatorieae, Asteraceae).

    PubMed

    Fernandes, Yanne S; Trindade, Luma M P; Rezende, Maria Helena; Paula, José R; Gonçalves, Letícia A

    2016-03-01

    Trichogonia cinerea is endemic to Brazil and occurs in areas of cerrado and campo rupestre. In this study, we characterized the glandular and non-glandular trichomes on the aerial parts of this species, determined the principal events in the development of the former, and identified the main constituents of the volatile oil produced in its aerial organs. Fully expanded leaves, internodes, florets, involucral bracts, and stem apices were used for the characterization of trichomes. Leaves, internodes, florets, and involucral bracts were examined by light microscopy and scanning electron microscopy, whereas stem apices were examined only by light microscopy. Branches in the reproductive phase were used for the extraction and determination of the composition of the volatile oil. The species has three types of glandular trichomes, biseriate vesicular, biseriate pedunculate, and multicellular uniseriate, which secrete volatile oils and phenolic compounds. The major components identified in the volatile oil were 3,5-muuroladiene (39.56%) and butylated hydroxytoluene (13.07%).

  17. Cleaning Effectiveness of a Reciprocating Single-file and a Conventional Rotary Instrumentation System.

    PubMed

    de Carvalho, Fredson Marcio Acris; Gonçalves, Leonardo Cantanhede de Oliveira; Marques, André Augusto Franco; Alves, Vanessa; Bueno, Carlos Eduardo da Silveira; De Martin, Alexandre Sigrist

    2016-01-01

    To compare cleaning effectiveness by histological analysis of a reciprocating single-file system with ProTaper rotary instruments during the preparation of curved root canals in extracted teeth. A total of 40 root canals with curvatures ranging between 20 - 40 degrees were divided into two groups of 20 canals. Canals were prepared to the following apical sizes: Reciproc size 25 (n=20); ProTaper: F2 (n=20). The normal distribution of data was tested by the Kolmogorov-Smirnov test and the values obtained for the test (Mann-Whitney U test, P < .05) were statistically analyzed using the GraphPad InStat for the Mac OS software (GraphPad Software, La Jolla, CA, USA). There were no significant differences in remaining debris (P > .05) between the two groups. The application of reciprocating motion during instrumentation did not result in increased debris when compared with continuous rotation motion, even in the apical part of curved canals. Both instruments resulted in debris in the canal lumen, irrespective of the movement kinematics applied.

  18. The Impact of Apical Patency in the Success of Endodontic Treatment of Necrotic Teeth with Apical Periodontitis: A Brief Review.

    PubMed

    Machado, Ricardo; Ferrari, Carlos Henrique; Back, Eduardo; Comparin, Daniel; Tomazinho, Luiz Fernando; Vansan, Luiz Pascoal

    2016-01-01

    Accumulation of soft tissue or dentinal remnants in the apical region is a common event that can cause blockage of root canals. This event can be avoided if apical patency is performed during the root canal shaping procedures. However, there is no consensus on the role of apical patency in relation to the success of endodontic treatment of necrotic teeth with apical periodontitis. Therefore, the purpose of this paper was to conduct a brief review on the role of apical patency in guaranteeing the success of endodontic treatments of necrotic teeth with apical periodontitis considering two other key points; the root canal anatomy and microbiology.

  19. Apical root resorption in maxillary incisors when employing micro-implant and J-hook headgear anchorage: a 4-month radiographic study.

    PubMed

    Wang, Qingzhu; Chen, Wenjing; Smales, Roger J; Peng, Hui; Hu, Xiaokun; Yin, Lu

    2012-10-01

    This study evaluated, over a 4-month study period, the amount of apical root resorption occurring in maxillary central incisors following their retraction when employing either micro-implant or J-hook headgear anchorage. The prospective randomised clinical trial was conducted in Orthodontic Clinic, College of Stomatology, China from 2008-2009. Subjects are patients requiring fixed appliances on waiting list (n=20). In female Han Chinese patients aged from 16-26 years, standardized periapical radiographs from 10 randomly assigned patients with maxillary protrusions comprising the micro-implant group, and from 10 similar patients comprising the J-hook headgear group, were assessed for maxillary central incisor apical root resorption. Measurements before and after orthodontic therapy were also obtained from lateral cephalometric radiographs to calculate incisor horizontal retraction and vertical intrusion distances. Estimated retraction force vectors were calculated in horizontal and vertical directions for both treatment groups. Data analysis employed t-tests and the Pearson correlation test, with α=0.05 for statistical significance. The results showed that when compared with the J-hook group, significantly more apical root resorption shortening of the maxillary central incisors was observed in the micro-implant group (1.27 mm difference, 95% CI=0.70-1.84, P<0.001), which was associated with a significantly larger retraction distance (P=0.004) and a smaller vertical force component (P<0.0001). We are led to conclude that continuous activation of the nickel-titanium coil springs used in the micro-implant group resulted in significantly more apical root resorption shortening and maxillary central incisor retraction than when intermittent J-hook retraction was employed. The employment of continuous duration orthodontic forces presents a risk for increased apical root resorption that requires careful radiographic monitoring.

  20. Bradykinin regulates human colonic ion transport in vitro

    PubMed Central

    Baird, A W; Skelly, M M; O'Donoghue, D P; Barrett, K E; Keely, S J

    2008-01-01

    Background and purpose: Kinins are acknowledged as important regulators of intestinal function during inflammation; however, their effects on human intestinal ion transport have not been reported. Here, we used muscle-stripped human colonic tissue and cultured T84-cell monolayers to study bradykinin (BK) actions on human intestinal ion transport. Experimental approach: Ion transport was measured as changes in short-circuit current (Isc) across colonic epithelia mounted in Ussing chambers. Key results: In intact tissue, there was a distinct polarity to BK-elicited Isc responses. Whereas basolateral BK stimulated sustained responses (EC50=0.5±0.1 μM), those to apical BK were more rapid and transient (EC50=4.1±1.2 nM). In T84 cells, responses to both apical and basolateral BK were similar to those seen upon apical addition to intact tissues. Cross-desensitization between apical and basolateral domains was not observed. BK-induced responses were largely due to Cl− secretion as shown by their sensitivity to bumetanide and removal of Cl− from the bathing solution. Studies using selective agonists and antagonists indicate responses to BK are mediated by B2 receptors. Finally, responses to basolateral BK in intact tissues were inhibited by tetrodotoxin (1 μM), atropine (1 μM), capsaicin (100 μM) and piroxicam (10 μM). BK-stimulated prostaglandin (PG)E2 release from colonic tissue. Conclusions: BK stimulates human colonic Cl− secretion by activation of apical and basolateral B2 receptors. Responses to apical BK reflect a direct action on epithelial cells, whereas those to basolateral BK are amplified by stimulation of enteric nerves and PG synthesis. PMID:18604228

  1. The GTP-bound and Sumoylated Form of the rab17 Small Molecular Weight GTPase Selectively Binds Syntaxin 2 in Polarized Hepatic WIF-B Cells*

    PubMed Central

    Striz, Anneliese C.; Tuma, Pamela L.

    2016-01-01

    A major focus for our laboratory is identifying the molecules and mechanisms that regulate polarized apical protein sorting in hepatocytes, the major epithelial cells of the liver. These trafficking pathways are regulated, in part, by small molecular weight rab GTPases. We chose to investigate rab17, whose expression is restricted to polarized epithelial cells, is enriched in liver, and has been implicated in regulating basolateral to apical transcytosis. To initiate our studies, we generated three recombinant adenoviruses expressing wild type, constitutively active (GTP bound), or dominant-negative (GDP bound) rab17. Immunoblotting revealed rab17 immunoreactive species at 25 kDa (the predicted rab17 molecular mass) and 40 kDa. We determined that mono-sumoylation of the 25-kDa rab17 is responsible for the shift in molecular mass, and that rab17 prenylation is required for sumoylation. We further determined that sumoylation selectively promotes interactions with syntaxin 2 (but not syntaxins 3 or 4) and that these interactions are nucleotide dependent. Furthermore, a K68R-mutated rab17 led to the redistribution of syntaxin 2 and 5′ nucleotidase from the apical membrane to subapical puncta, whereas multidrug resistance protein 2 distributions were not changed. Together these data are consistent with the proposed role of rab17 in vesicle fusion with the apical plasma membrane and further implicate sumoylation as an important mediator of protein-protein interactions. The selectivity in syntaxin binding and apical protein redistribution further suggests that rab17 and syntaxin 2 mediate fusion of transcytotic vesicles at the apical surface. PMID:26957544

  2. 75 FR 75934 - Airworthiness Directives; Apical Industries Inc. (Apical) Emergency Float Kits

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-07

    ...-1190; Directorate Identifier 2010-SW-038-AD] RIN 2120-AA64 Airworthiness Directives; Apical Industries Inc. (Apical) Emergency Float Kits AGENCY: Federal Aviation Administration, DOT. ACTION: Notice of... the Apical emergency float kits installed on certain model helicopters under supplemental type...

  3. Micro-CT evaluation of apical delta morphologies in human teeth.

    PubMed

    Gao, Xianhua; Tay, Franklin R; Gutmann, James L; Fan, Wei; Xu, Ting; Fan, Bing

    2016-11-07

    The apical delta is an intricate system within the root canal and incompletely debridement may affect the long-term prognosis of root canal therapy. The aim of the present study is to investigate the morphologic features of apical deltas in human teeth with micro-computed tomography (micro-CT) using a centreline-fitting algorithm. One hundred and thirty-six apical deltas were detected in 1400 teeth. Molars had more apical deltas (15.8%) than anterior teeth (6.3%). In maxillary molars, the mesiobuccal root had a significantly higher prevalence of apical delta than the palatal root or the distobuccal root. The median vertical distance of the apical delta was 1.87 mm with 13% more than 3 mm. The median diameter and length of the apical delta branches were 132.3 and 934.5 μm. Apical delta branches were not straight with cross-sectional shapes being non-circular. These morphological features of apical delta may complicate debridement of the infected root canal system.

  4. Trafficking Ion Transporters to the Apical Membrane of Polarized Intestinal Enterocytes.

    PubMed

    Engevik, Amy Christine; Goldenring, James R

    2018-01-02

    Epithelial cells lining the gastrointestinal tract require distinct apical and basolateral domains to function properly. Trafficking and insertion of enzymes and transporters into the apical brush border of intestinal epithelial cells is essential for effective digestion and absorption of nutrients. Specific critical ion transporters are delivered to the apical brush border to facilitate fluid and electrolyte uptake. Maintenance of these apical transporters requires both targeted delivery and regulated membrane recycling. Examination of altered apical trafficking in patients with Microvillus Inclusion disease caused by inactivating mutations in MYO5B has led to insights into the regulation of apical trafficking by elements of the apical recycling system. Modeling of MYO5B loss in cell culture and animal models has led to recognition of Rab11a and Rab8a as critical regulators of apical brush border function. All of these studies show the importance of apical membrane trafficking dynamics in maintenance of polarized epithelial cell function. Copyright © 2018 Cold Spring Harbor Laboratory Press; all rights reserved.

  5. Long-term prognosis of endodontically treated teeth: a retrospective analysis of preoperative factors in molars.

    PubMed

    Setzer, Frank C; Boyer, Keith R; Jeppson, Joshua R; Karabucak, Bekir; Kim, Syngcuk

    2011-01-01

    Long-term predictability of restored endodontically treated teeth is important for the decision of tooth retention versus extraction and implant placement. The purpose of this study was to validate the hypothesis that preoperative factors can predict the long-term prognosis of molars requiring endodontic and restorative treatment for future prognostic investigations. A clinical database was searched for molar endodontic treatments with crown placement and a minimum of 4-year follow-up. Charts of 42 patients with 50 individual treatments were randomly selected. Information concerning crown lengthening; periodontal diagnosis; attachment loss; furcation involvement; mobility; and internal, external, or periradicular resorption was recorded. Radiographs from treatment initiation and follow-up were digitalized. The presence of apical periodontitis was evaluated. Available ferrule was calculated from bitewing radiographs using CAD software (AutoCAD; Autodesk, Cupertino, CA). The resulting data, age, sex, and times of restoration and follow-up were analyzed for correlation with the presence of apical radiolucency at follow-up and the following four possible outcome scenarios: "no event," "nonsurgical retreatment," "surgical retreatment," or "extraction" using Spearman rank order correlation analysis. Patients' ages ranged from 19 to 87 years, 22 were male and 20 female, and 48 teeth (96.0%) were retained at follow-up. Of those, 44 (88.0%) were without intervention ("no event"), and four (8.0%) underwent surgical or nonsurgical retreatment. Two teeth (4.0%) had been extracted. Significant positive correlations existed between "untoward events" (any form of retreatment or extraction) and "prognostic value according to periodontal status" (p = 0.047) and "attachment loss" (p = 0.042). The only preoperative factors significant for the prognosis of restored endodontically treated molars were related to periodontal prognostic value and attachment loss. It can be concluded that it may be difficult to predict the prognosis of molars in need for endodontic treatment and restoration from prognostic factors not related to periodontal disease. Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  6. Further characterization of a furanocoumarin-free grapefruit juice on drug disposition: studies with cyclosporine.

    PubMed

    Paine, Mary F; Widmer, Wilbur W; Pusek, Susan N; Beavers, Kimberly L; Criss, Anne B; Snyder, Jennifer; Watkins, Paul B

    2008-04-01

    We previously established furanocoumarins as mediators of the interaction between grapefruit juice (GFJ) and the model CYP3A4 substrate felodipine in healthy volunteers using a GFJ devoid of furanocoumarins. It remains unclear whether furanocoumarins mediate drug-GFJ interactions involving CYP3A4 substrates that are also P-glycoprotein substrates. The effects of furanocoumarin-free GFJ on drug disposition were further characterized by using the dual CYP3A4/P-glycoprotein substrate cyclosporine. By randomized crossover design, 18 healthy volunteers received cyclosporine (5 mg/kg) with 240 mL orange juice (control), GFJ, or furanocoumarin-free GFJ. Blood was collected over 24 h. Juice treatments were separated by > or = 1 wk. The effects of diluted extracts of each juice and of purified furanocoumarins on [3H]cyclosporine translocation in Caco-2 cells were then compared. The median (range) dose-corrected cyclosporine area under the curve and the maximum concentration with GFJ (P < or = 0.007), but not with furanocoumarin-free GFJ (P > or = 0.50), were significantly higher than those with orange juice [15.6 (6.7-33.5) compared with 11.3 (4.8-22.0) x 10(-3) h/L and 3.0 (1.6-5.8) compared with 2.4 (1.1-3.1) mL(-1), respectively]. The median time to reach maximum concentration and terminal elimination half-life were not significantly different between the juices (2-3 and 7-8 h, respectively; P > or = 0.08). Relative to vehicle, the GFJ extract, orange juice extract, and purified furanocoumarins partially increased apical-to-basolateral and decreased basolateral-to-apical [3H]cyclosporine translocation in Caco-2 cells, whereas the furanocoumarin-free GFJ extract had negligible effects. Reanalysis of the clinical juices identified polymethoxyflavones as candidate P-glycoprotein inhibitors in orange juice but not in GFJ. Furanocoumarins mediate, at least partially, the cyclosporine-GFJ interaction in vivo. A plausible mechanism involves the combined inhibition of enteric CYP3A4 and P-glycoprotein.

  7. [Effects of different techniques on removal of vapor lock in the apical region of curved canals: a cone-beam computed tomography study].

    PubMed

    Su, Z; Bai, Y H; Hou, X M

    2017-02-18

    To compare the effect of four different techniques on removal of vapor lock in the apical region of curved root canals. Forty simulated resin root canals with 45° curvature were prepared using WaveOne Primary, then the apical foramen were sealed with soft wax. The teeth were divided randomly into 4 groups thereafter (n=10). Contract solution was injected into the canals using a 30 G side-vented needle and scanned with cone-beam CT (CBCT) to identify the volume of the vapor lock. Four different techniques including photon-induced photoacoustic streaming (PIPS) laser-activated irrigation, gutta-percha cone technique, ultrasonic irrigation, and sonic irrigation were used to remove the vapor locks in the root canals. The residual volume of the vapor lock was identified again using CBCT scanning data. Accordingly, the reduction rates of the vapor lock were calculated. Furthermore, the initial and residual vapor lock length was calculated. The data were analyzed by using the One-way ANOVA analysis and Kruskal-Wallis H test at a significance level of P<0.05. There was no significant difference in the initial vapor lock volume (P>0.05). Residual volume of the vapor lock for PIPS laser-activated irrigation was 0 mm(3), and that for gutta-percha cone technique was (0.02±0.07) mm3, significantly lower than those of ultrasonic and sonic irrigation, the values being (0.20±0.09) mm(3) and (0.23±0.06) mm(3) (P<0.001), respectively. The reduction rates of the vapor lock of PIPS laser-activated irrigation and gutta-percha cone technique were 100.00% (100.00%, 100.00%) and 100.00% (77.66%, 100.00%), respectively, significantly higher than those of ultrasonic irrigation [70.37% (56.41%, 91.43%)] and sonic irrigation [63.54% (51.47%, 74.00%), P<0.001]. The length of the residual vapor lock for PIPS laser-activated irrigation was 0 mm, and that for gutta-percha cone technique was (0.15±0.47) mm, significantly lower than those of ultrasonic and sonic irrigation, values being (2.21±0.09) mm and (2.34±0.08) mm (P<0.001), respectively. The length of the residual vapor locks in the ultrasonic and sonic group remained approximately the same as the distance between the working tip and the apical foramen. PIPS laser activated irrigation and gutta-percha cone technique could remove the vapor lock from the apical region of curved canals effectively.

  8. Aldosterone regulation of sodium and potassium transport in the cortical collecting duct.

    PubMed

    O'Neil, R G

    1990-07-01

    The aldosterone-induced up-regulation of Na absorption and K secretion in the CCD is complex and involves the regulation of numerous transport proteins. Some aspects of the response may be species dependent. For example, stimulation of Na and K transport in the rabbit CCD involves a marked up-regulation in the apical cell membrane Na and K conductances, the basolateral cell membrane K conductance, and the basolateral membrane NaK-ATPase activity. In the rat CCD, aldosterone causes a similar up-regulation in the NaK-ATPase and the apical membrane Na conductance, but supposedly has little influence on the apical and basolateral membrane K conductances as evaluated by indirect methods. Furthermore, the marked hyperpolarization of the basolateral membrane with long-term aldosterone treatment in the rabbit CCD is blunted or absent in the rat CCD. Other differences between the CCD of these two species have been outlined. Nonetheless, the basic responses of the CCDs from the two species show similar trends. The actions of aldosterone in the CCD principal cell are summarized in Figure 5. The initial steps have been described previously. Aldosterone (A) diffuse across the cell membrane and binds to a cytoplasmic receptor (R). The receptor complex moves into the nucleus and binds to an acceptor site on chromatin, initiating transcription and the subsequent synthesis of a myriad of new proteins referred to as aldosterone-induced proteins (AIP). The initial observed action of aldosterone is an upregulation of the apical membrane Na conductance during the early phase, which occurs within 1 to 2 hours. The increase in Na conductance likely reflects activation of preexisting latent Na channels and not synthesis of new channels, although activation does require protein synthesis. The increased Na influx during the early phase presents a larger Na load to the Na pump, which is likely reflected as a modest transient increase in intracellular Na activity. Based on kinetic considerations alone, this should cause an increased transport turnover of the pump with a greater Na extrusion rate and K uptake rate. The stimulated Na influx also causes a modest depolarization of the apical membrane during the early phase, which when combined with the increased K uptake via the pump and an apparent modest elevation in the intracellular K activity, results in a more favorable gradient for K secretion (increased driving force) into the tubule lumen.(ABSTRACT TRUNCATED AT 400 WORDS)

  9. Impacts of oxidative stress on acetylcholinesterase transcription, and activity in embryos of zebrafish (Danio rerio) following Chlorpyrifos exposure.

    PubMed

    Rodríguez-Fuentes, Gabriela; Rubio-Escalante, Fernando J; Noreña-Barroso, Elsa; Escalante-Herrera, Karla S; Schlenk, Daniel

    2015-01-01

    Organophosphate pesticides cause irreversible inhibition of AChE which leads to neuronal overstimulation and death. Thus, dogma indicates that the target of OP pesticides is AChE, but many authors postulate that these compounds also disturb cellular redox processes, and change the activities of antioxidant enzymes. Interestingly, it has also been reported that oxidative stress plays also a role in the regulation and activity of AChE. The aims of this study were to determine the effects of the antioxidant, vitamin C (VC), the oxidant, t-butyl hydroperoxide (tBOOH) and the organophosphate Chlorpyrifos (CPF), on AChE gene transcription and activity in zebrafish embryos after 72h exposure. In addition, oxidative stress was evaluated by measuring antioxidant enzymes activities and transcription, and quantification of total glutathione. Apical effects on the development of zebrafish embryos were also measured. With the exception of AChE inhibition and enhanced gene expression, limited effects of CPF on oxidative stress and apical endpoints were found at this developmental stage. Addition of VC had little effect on oxidative stress or AChE, but increased pericardial area and heartbeat rate through an unknown mechanism. TBOOH diminished AChE gene expression and activity, and caused oxidative stress when administered alone. However, in combination with CPF, only reductions in AChE activity were observed with no significant changes in oxidative stress suggesting the adverse apical endpoints in the embryos may have been due to AChE inhibition by CPF rather than oxidative stress. These results give additional evidence to support the role of prooxidants in AChE activity and expression. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Analysis of Aprotinin, a Protease Inhibitor, Action on the Trafficking of Epithelial Na+ Channels (ENaC) in Renal Epithelial Cells Using a Mathematical Model.

    PubMed

    Sasamoto, Kouhei; Marunaka, Rie; Niisato, Naomi; Sun, Hongxin; Taruno, Akiyuki; Pezzotti, Giuseppe; Yamamoto, Toshiro; Kanamura, Narisato; Zhu, Wenliang; Nishio, Kyosuke; Inui, Toshio; Eaton, Douglas C; Marunaka, Yoshinori

    2017-01-01

    Epithelial Na+ channels (ENaC) play a crucial role in control of blood pressure by regulating renal Na+ reabsorption. Intracellular trafficking of ENaC is one of the key regulators of ENaC function, but a quantitative description of intracellular recycling of endogenously expressed ENaC is unavailable. We attempt here to provide a model for intracellular recycling after applying a protease inhibitor under hypotonic conditions. We simulated the ENaC-mediated Na+ transport in renal epithelial A6 cells measured as short-circuit currents using a four-state mathematical ENaC trafficking model. We developed a four-state mathematical model of ENaC trafficking in the cytosol of renal epithelial cells that consists of: an insertion state of ENaC that can be trafficked to the apical membrane state (insertion rate); an apical membrane state of ENaC conducting Na+ across the apical membrane; a recycling state containing ENaC that are retrieved from the apical membrane state (endocytotic rate) and then to the insertion state (recycling rate) communicating with the apical membrane state or to a degradation state (degradation rate). We studied the effect of aprotinin (a protease inhibitor) blocking protease-induced cleavage of the extracellular loop of γ ENaC subunit on the rates of intracellular ENaC trafficking using the above-defined four-state mathematical model of ENaC trafficking and the recycling number relative to ENaC staying in the apical membrane. We found that aprotinin significantly reduced the insertion rate of ENaC to the apical membrane by 40%, the recycling rate of ENaC by 81%, the cumulative time of an individual ENaC staying in the apical membrane by 32%, the cumulative life-time after the first endocytosis of ENaC by 25%, and the cumulative Na+ absorption by 31%. The most interesting result of the present study is that cleavage of ENaC affects the intracellular ENaC trafficking rate and determines the residency time of ENaC, indicating that more active cleaved ENaCs stay longer at the apical membrane contributing to transcellular Na+ transport via an increase in recycling of ENaC to the apical membrane. The extracellular protease-induced cleavage of the extracellular loop of γ ENaC subunit increases transcellular epithelial Na+ transport by elevating the recycling rate of ENaC due to an increase in the recycling rate of ENaCs associated with increases in the insertion rate of ENaC. © 2017 The Author(s). Published by S. Karger AG, Basel.

  11. Modification and validation of a high-performance liquid chromatography method for quantification of Huperzine A in Huperzia crispata.

    PubMed

    Yu, Lijun; Shi, Yunfeng; Huang, Jianan; Gong, Yushun; Liu, Zhonghua; Hu, Weixin

    2010-01-01

    The present study describes a rapid and sensitive HPLC method for the quantification of huperzine A (HupA) in Huperzia crispata (Huperziaceae). The sample extraction and preparation involved a simple, time-saving, single-solvent extraction, with each sample being analyzed within 12 min. The mobile phase was ammonium acetate (0.1 M, pH 6.0)--methanol (64 + 36, v/v) at a flow rate of 1.0 mL/min. Detection was at 308 nm. The calibration curve was linear from 0.049 to 7.84 microg (R2 = 0.9997), with intraday and interday precision RSD of less than 2%. The extraction recovery rate was over 98.49%. Quantification of HupA was performed using this modified method, and the content of HupA was 1.86 times higher in the whole plant of H. crispata (218.17 +/- 1.55 microg/g) than in that of H. serrata (117.03 +/- 2.97 microg/g). In the whole plant of H. crispata, HupA mainly accumulated in the actively growing shoot tips, the apical bud, and the 10 youngest leaves, reaching 455.23 +/- 2.97 microg/g. The content of HupA in the samples from sunshine-sheltered sites was 3.45 times higher than in that from sunshine-abundant sites. The satisfactory results indicate that this modified method can be applied in the quality control of large-scale Huperziaceae plant extracts and that changes should be made in the cultivation of H. crispata so as to maximize the production of HupA.

  12. Relationship between human cytomegalovirus transcription and symptomatic apical periodontitis in Iran.

    PubMed

    Yazdi, K A; Sabeti, M; Jabalameli, F; Eman eini, M; Kolahdouzan, S A; Slots, J

    2008-12-01

    Apical periodontitis of endodontic origin may develop as a result of cooperative interactions among herpesviruses, specific pathogenic bacteria and tissue-destructive inflammatory mediators. This study sought to identify the presence of Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) transcripts in symptomatic and asymptomatic periapical lesions of individuals living in Iran. Fifty endodontic patients (28 with symptomatic periapical lesions and 22 with asymptomatic periapical lesions) were included in the study. In each study subject, a microbiological periapical sample was collected using a curette in conjunction with periapical surgery. A reverse transcription-polymerase chain reaction assay was used to identify transcripts of EBV and HCMV. Human cytomegalovirus transcript was detected in 15 of the 28 (53.6%) symptomatic and in six of the 22 (27.3%) asymptomatic periapical study lesions (significant difference between symptomatic and asymptomatic lesions; P = 0.03, chi-square test). Epstein-Barr virus transcript was identified in one symptomatic and in two asymptomatic periapical lesions. This study establishes that HCMV transcription is common in apical periodontitis and is most frequent in symptomatic lesions. The high frequency of active herpesvirus infections in severe apical periodontitis changes the pathogenic paradigm of the disease and may also have preventive and therapeutic implications.

  13. Antioxidant Activity of a Geopropolis from Northeast Brazil: Chemical Characterization and Likely Botanical Origin.

    PubMed

    Ferreira, Joselena M; Fernandes-Silva, Caroline C; Salatino, Antonio; Message, Dejair; Negri, Giuseppina

    2017-01-01

    Geopropolis is a product containing wax, plant resin, and soil particles. It is elaborated by stingless bees of tribe Meliponini. Methanol extracts of sample of geopropolis produced by Scaptotrigona postica ("mandaguari") in the state of Rio Grande do Norte (RN, northeast Brazil) were analyzed for the determination of standard parameters (total phenols, total flavonoids, and radical scavenging activity) and chemical characterization by HPLC-DAD-MS/MS analysis. The sample analyzed has high contents of total phenols and flavonoids, as well as high antioxidant activity. The constituents characterized were mainly flavonols, such as quercetin methyl ethers, and methoxychalcones. Such chemical profile is similar to the composition of a green propolis from the same area of RN, which is produced by Africanized Apis mellifera , using shoot apices of Mimosa tenuiflora , popularly known as "jurema-preta." This finding provides evidence that "mandaguari" geopropolis and honeybee propolis have the same botanical origin in RN. The sharing of a plant resin source by phylogenetically distant bees (Apinae and Meliponinae) suggests that bee genetic factors play little role in the choice of plants for resin collection and that the availability of potential botanical sources plays a decisive role.

  14. Antioxidant Activity of a Geopropolis from Northeast Brazil: Chemical Characterization and Likely Botanical Origin

    PubMed Central

    Ferreira, Joselena M.; Message, Dejair

    2017-01-01

    Geopropolis is a product containing wax, plant resin, and soil particles. It is elaborated by stingless bees of tribe Meliponini. Methanol extracts of sample of geopropolis produced by Scaptotrigona postica (“mandaguari”) in the state of Rio Grande do Norte (RN, northeast Brazil) were analyzed for the determination of standard parameters (total phenols, total flavonoids, and radical scavenging activity) and chemical characterization by HPLC-DAD-MS/MS analysis. The sample analyzed has high contents of total phenols and flavonoids, as well as high antioxidant activity. The constituents characterized were mainly flavonols, such as quercetin methyl ethers, and methoxychalcones. Such chemical profile is similar to the composition of a green propolis from the same area of RN, which is produced by Africanized Apis mellifera, using shoot apices of Mimosa tenuiflora, popularly known as “jurema-preta.” This finding provides evidence that “mandaguari” geopropolis and honeybee propolis have the same botanical origin in RN. The sharing of a plant resin source by phylogenetically distant bees (Apinae and Meliponinae) suggests that bee genetic factors play little role in the choice of plants for resin collection and that the availability of potential botanical sources plays a decisive role. PMID:29234387

  15. Revascularization and periapical repair after endodontic treatment using apical negative pressure irrigation versus conventional irrigation plus triantibiotic intracanal dressing in dogs' teeth with apical periodontitis.

    PubMed

    da Silva, Lea Assed Bezerra; Nelson-Filho, Paulo; da Silva, Raquel Assed Bezerra; Flores, Daniel Silva Herzog; Heilborn, Carlos; Johnson, James D; Cohenca, Nestor

    2010-05-01

    The objective of this study was to evaluate in vivo the revascularization and the apical and periapical repair after endodontic treatment using 2 techniques for root canal disinfection (apical negative pressure irrigation versus apical positive pressure irrigation plus triantibiotic intracanal dressing) in immature dogs' teeth with apical periodontitis. Two test groups of canals with experimentally induced apical periodontitis were evaluated according to the disinfection technique: Group 1, apical negative pressure irrigation (EndoVac system), and Group 2, apical positive pressure irrigation (conventional irrigation) plus triantibiotic intracanal dressing. In Group 3 (positive control), periapical lesions were induced, but no endodontic treatment was done. Group 4 (negative control) was composed of sound teeth. The animals were killed after 90 days and the maxillas and mandibles were subjected to histological processing. The sections were stained with hematoxylin and eosin and Mallory Trichrome and examined under light microscopy. A description of the apical and periapical features was done and scores were attributed to the following histopathological parameters: newly formed mineralized apical tissue, periapical inflammatory infiltrate, apical periodontal ligament thickness, dentin resorption, and bone tissue resorption. Intergroup comparisons were done by the Kruskal-Wallis and Dunn's tests (alpha = 0.05). Although statistically significant difference was found only for the inflammatory infiltrate (P < .05), Group 1 presented more exuberant mineralized formations, more structured apical and periapical connective tissue, and a more advanced repair process than Group 2. From the histological observations, sodium hypochlorite irrigation with the EndoVac system can be considered as a promising disinfection protocol in immature teeth with apical periodontitis, suggesting that the use of intracanal antibiotics might not be necessary. Copyright (c) 2010 Mosby, Inc. All rights reserved.

  16. The cuticular localization of integument peptides from particular routing categories.

    PubMed

    Locke, M; Kiss, A; Sass, M

    1994-10-01

    The distribution of integument peptides in relation to chitin and structural features has been studied in the surface epidermis of the caterpillar of Calpodes ethlius by immunoblotting and immunogold labelling using antibodies prepared to peptides isolated from lamellate endocuticle or from hemolymph. The intermoult cuticle consists of an epicuticle, an endocuticle of many chitin containing lamellae, and a chitin containing assembly zone directly above the apical epidermal microvilli and the perimicrovillar space. During the intermoult, the epidermis secretes peptides constitutively, that is, secretory vesicles containing peptides exocytose without accumulating, traverse the perimicrovillar space and form lamellae in the assembly zone. At moulting, the epidermis deposits ecdysial droplets in addition. These interrupt the last few lamellae which later go on to become the perforated ecdysial membrane. The integument is involved with four routing classes of peptide. Secretion is apical into the cuticle (C), basal into the hemolymph (H), bidirectional (BD), or transported to the cuticle across the epidermis from the hemolymph (T). Some peptides change their routing at moulting. There are several patterns of localization. (1) C and BD cuticular peptides occur mainly in chitin containing lamellate cuticle. (2) Some are also present in epicuticle, and are therefore not obligatorily linked to chitin or matrix between chitin fibers. Cuticular peptides that also occur in the hemolymph are glycosylated, whereas most that are only secreted apically into the cuticle are not. All BD but few C peptides carry alpha-D-glucose/alpha-D-mannose. Some C and BD peptides carry N-acetyl glucosamine. (3) C36 extracted from cuticle has most N-acetyl glucosamine and colocalizes with chitin rather than the protein matrix. It is therefore probably the main link between chitin fibers and the matrix. (4) H235 is barely detectable at the apical cell surface during the intermoult but is abundant at moulting around and below the ecdysial droplets. (5) T66 occurs in intermoult lamellate cuticle. At moulting, alone among the peptides examined, it is in ecdysial droplets. Intermoult C and BD peptides are not in ecdysial droplets but continue to be present in the ecdysial membrane, suggesting that constitutive secretion is independent from the exocytosis of transported moult peptides. T66 differs from most hemolymph peptides in that it does not carry N-acetyl glucosamine or alpha-D-glucose/alpha-D-mannose. (6) Weakly reacting BD peptides (and some H peptides barely detectable in cuticle) localize near the apical surface. Their distribution therefore favours apical secretion and retrieval as a mechanism for basal secretion.

  17. Apical Correlative Effects in Leaf Epinasty of Tomato 1

    PubMed Central

    Kazemi, Saidollah; Kefford, Noel P.

    1974-01-01

    The influence of the stem apex on leaf curvature was investigated using debudded tomato (Lycopersicon esculentum Mill. cv Anahu) plants and petiole explants, consisting of a section of petiole attached to a section of stem. Decapitation of the main shoot of tomato plants induced hyponasty of petioles in young leaves. Application of auxin in place of the removed apex or fumigation of intact tomato plants with ethylene produced epinastic curvature at the base of the petiole. Simultaneous carbon dioxide treatments prevented the development of petiolar epinasty due to auxin and ethylene treatments. Application of ethylene gas to the decapitated shoot or injection into the stem, induced petiolar epinasty. In a saturating level of ethylene gas, tomato petioles did not respond to indole-3-acetic acid applied to the cut apex. Auxin-induced ethylene production in petiole explants preceded the development of epinasty. Application of indoleacetic acid in lanolin to the entire lower side of the petioles of leaves in situ produced petiole epinasty. Petiolar epinasty due to apically applied indoleacetic acid resulted from differential cell elongation. The auxins indole-3-acetic acid, 2,4-dichlorophenoxyacetic acid, and naphthalene-1-acetic acid induced epinasty when applied apically to decapitated tomato plants, while gibberellic acid, kinetin, abscisic acid, and auxin or gibberellin antagonists had no effect. When such compounds were applied to petiole explants, only indole-3-acetic acid and kinetin caused an increase in ethylene production and the effect of kinetin was relatively weak. Application of 2,3,5-triiodobenzoic acid around the stem did not change the effect on petiolar epinasty of auxin applied to the decapitated shoot or around the stem. Radioautography showed that the label from 14C-indoleacetic acid applied apically entered the petiole and midrib tissue; however, extraction showed that only a fraction of the label in these tissues was in the form of indoleacetic acid. Removal of leaflets from leaves induced hyponasty in the midrib region, and application of auxin to the leaflet stubs produced midrib epinasty; carbon dioxide did not block the action of auxin in this type of epinasty. Removal of leaflets from leaves did not alter the effect of apically applied auxin on petiolar epinasty. The data are consistent with the hypothesis that the oblique orientation of leaves in tomato plants is influenced by two epinastic responses. Petiolar epinasty is controlled by the apical region on the stem and is due to the action of auxin-induced ethylene; and midrib epinasty is due to an action of auxin other than through ethylene. PMID:16658919

  18. The apical complex couples cell fate and cell survival to cerebral cortical development

    PubMed Central

    Kim, Seonhee; Lehtinen, Maria K.; Sessa, Alessandro; Zappaterra, Mauro; Cho, Seo-Hee; Gonzalez, Dilenny; Boggan, Brigid; Austin, Christina A.; Wijnholds, Jan; Gambello, Michael J.; Malicki, Jarema; LaMantia, Anthony S.; Broccoli, Vania; Walsh, Christopher A.

    2010-01-01

    Cortical development depends upon tightly controlled cell fate and cell survival decisions that generate a functional neuronal population, but the coordination of these two processes is poorly understood. Here we show that conditional removal of a key apical complex protein, Pals1, causes premature withdrawal from the cell cycle, inducing excessive generation of early-born postmitotic neurons followed by surprisingly massive and rapid cell death, leading to the abrogation of virtually the entire cortical structure. Pals1 loss shows exquisite dosage sensitivity, so that heterozygote mutants show an intermediate phenotype on cell fate and cell death. Loss of Pals1 blocks essential cell survival signals, including the mammalian target of rapamycin (mTOR) pathway, while mTORC1 activation partially rescues Pals1 deficiency. These data highlight unexpected roles of the apical complex protein Pals1 in cell survival through interactions with mTOR signaling. PMID:20399730

  19. Regulation of podocalyxin trafficking by Rab small GTPases in epithelial cells

    PubMed Central

    Mrozowska, Paulina S.; Fukuda, Mitsunori

    2016-01-01

    ABSTRACT The characteristic feature of polarity establishment in MDCK II cells is transcytosis of apical glycoprotein podocalyxin (PCX) from the outer plasma membrane to the newly formed apical domain. This transcytotic event consists of multiple steps, including internalization from the plasma membrane, transport through early endosomes and Rab11-positive recycling endosomes, and delivery to the apical membrane. These steps are known to be tightly coordinated by Rab small GTPases, which act as molecular switches cycling between active GTP-bound and inactive GDP-bound states. However, our knowledge regarding which sets of Rabs regulate particular steps of PCX trafficking was rather limited. Recently, we have performed a comprehensive analysis of Rab GTPase engagement in the transcytotic pathway of PCX during polarity establishment in 2-dimensional (2D) and 3-dimensional (3D) MDCK II cell cultures. In this Commentary we summarize our findings and set them in the context of previous reports. PMID:27463697

  20. The zymographic evaluation of gelatinase (MMP-2 and -9) levels in acute and chronic periapical abscesses.

    PubMed

    Buzoglu, Hatice Dogan; Unal, Hasan; Ulger, Celal; Mert, Safak; Kücükyildirim, Sibel; Er, Nuray

    2009-11-01

    This study investigated the presence and levels of matrix metalloproteinases (MMP)-2 and -9 in periapical abscesses. Eighteen samples of intracanal exudates containing pus were collected from teeth with clinically and radiographically verified primary or secondary acute and chronic apical abscesses. Pro- and active forms of MMP-2 and MMP-9 levels were analyzed by using substrate gel zymography followed by an image analysis system. Statistical analysis was performed using the Kruskall-Wallis and Mann-Whitney U tests with Bonferroni adjustment. Both forms of MMP-9 were detected in all pus samples and demonstrated marked differences among the experimental groups (P < .05). Primary or secondary acute apical abscess samples demonstrated significantly higher MMP-9 levels compared with MMP-2 levels (P < .01). However, MMP-2 could not be detected in chronic apical abscesses. According to the results of this study, gelatinases might affect the pathogenesis of acute and chronic periapical abscesses.

  1. Actinomyces meyeri brain abscess following dental extraction

    PubMed Central

    Clancy, U; Ronayne, A; Prentice, M B; Jackson, A

    2015-01-01

    We describe the rare occurrence of an Actinomyces meyeri cerebral abscess in a 55-year-old woman following a dental extraction. This patient presented with a 2-day history of hemisensory loss, hyper-reflexia and retro-orbital headache, 7 days following a dental extraction for apical peridonitis. Neuroimaging showed a large left parietal abscess with surrounding empyema. The patient underwent craniotomy and drainage of the abscess. A. meyeri was cultured. Actinomycosis is a rare cause of cerebral abscess. The A. meyeri subtype is particularly rare, accounting for less than 1% of specimens. This case describes an unusually brief course of the disease, which is usually insidious. Parietal lobe involvement is unusual as cerebral abscesses usually have a predilection for the frontal and temporal regions of the brain. Although there are no randomised trials to guide therapy, current consensus is to use a prolonged course of intravenous antibiotics, followed by 6–12 months of oral therapy. PMID:25870213

  2. Smear Layer Outcome on Healing

    DTIC Science & Technology

    2015-07-31

    apical tissues, symptomatic apical periodontitis , asymptomatic apical periodontitis , chronic apical abscess or condensing osteomyelitis - Presence...percha • Patients presenting with periodontal disease or acute apical abscess ·Patients currently taking antibiotics If a patient expressed an... periodontitis : A randomized clinical trial. Joumal of Endodontics, 33, 1145-48. 23. Moss D, Allemang J, Johnson J. (2001). Philosophies and Practices

  3. RhoA regulates actin network dynamics during apical surface emergence in multiciliated epithelial cells

    PubMed Central

    Sedzinski, Jakub; Hannezo, Edouard; Tu, Fan; Biro, Maté

    2017-01-01

    ABSTRACT Homeostatic replacement of epithelial cells from basal precursors is a multistep process involving progenitor cell specification, radial intercalation and, finally, apical surface emergence. Recent data demonstrate that actin-based pushing under the control of the formin protein Fmn1 drives apical emergence in nascent multiciliated epithelial cells (MCCs), but little else is known about this actin network or the control of Fmn1. Here, we explore the role of the small GTPase RhoA in MCC apical emergence. Disruption of RhoA function reduced the rate of apical surface expansion and decreased the final size of the apical domain. Analysis of cell shapes suggests that RhoA alters the balance of forces exerted on the MCC apical surface. Finally, quantitative time-lapse imaging and fluorescence recovery after photobleaching studies argue that RhoA works in concert with Fmn1 to control assembly of the specialized apical actin network in MCCs. These data provide new molecular insights into epithelial apical surface assembly and could also shed light on mechanisms of apical lumen formation. PMID:28089989

  4. RhoA regulates actin network dynamics during apical surface emergence in multiciliated epithelial cells.

    PubMed

    Sedzinski, Jakub; Hannezo, Edouard; Tu, Fan; Biro, Maté; Wallingford, John B

    2017-01-15

    Homeostatic replacement of epithelial cells from basal precursors is a multistep process involving progenitor cell specification, radial intercalation and, finally, apical surface emergence. Recent data demonstrate that actin-based pushing under the control of the formin protein Fmn1 drives apical emergence in nascent multiciliated epithelial cells (MCCs), but little else is known about this actin network or the control of Fmn1. Here, we explore the role of the small GTPase RhoA in MCC apical emergence. Disruption of RhoA function reduced the rate of apical surface expansion and decreased the final size of the apical domain. Analysis of cell shapes suggests that RhoA alters the balance of forces exerted on the MCC apical surface. Finally, quantitative time-lapse imaging and fluorescence recovery after photobleaching studies argue that RhoA works in concert with Fmn1 to control assembly of the specialized apical actin network in MCCs. These data provide new molecular insights into epithelial apical surface assembly and could also shed light on mechanisms of apical lumen formation. © 2017. Published by The Company of Biologists Ltd.

  5. Subdural abscess secondary to covert dental sepsis

    PubMed Central

    Sprott, Mae S.; Hall, K.; Newman, P. K.; Welbury, R. R.; Ingham, H. R.

    1981-01-01

    The bacterial flora of a subdural abscess in a 17-year-old male, with radiological evidence of unilateral infection of the maxillary and frontal air sinuses, was typical of that encountered in the dental sulcus. Extensive examination revealed no primary focus of infection other than apical infection in the 2 upper first molar teeth, which were extracted. Treatment with ampicillin, gentamicin and metronidazole rapidly controlled the subdural infection, and resolution, as evidenced by computerized tomography, was complete at 10 months. PMID:7335566

  6. Mycoplasma detection and elimination are necessary for the application of stem cell from human dental apical papilla to tissue engineering and regenerative medicine.

    PubMed

    Kim, Byung-Chul; Kim, So Yeon; Kwon, Yong-Dae; Choe, Sung Chul; Han, Dong-Wook; Hwang, Yu-Shik

    2015-01-01

    Recently, postnatal stem cells from dental papilla with neural crest origin have been considered as one of potent stem cell sources in regenerative medicine regarding their multi-differentiation capacity and relatively easy access. However, almost human oral tissues have been reported to be infected by mycoplasma which gives rise to oral cavity in teeth, and mycoplasma contamination of ex-vivo cultured stem cells from such dental tissues and its effect on stem cell culture has received little attention. In this study, mycoplama contamination was evaluated with stem cells from apical papilla which were isolated from human third molar and premolars from various aged patients undergoing orthodontic therapy. The ex-vivo expanded stem cells from apical papilla were found to express stem cell markers such as Stro-1, CD44, nestin and CD133, but mycoplama contamination was detected in almost all cell cultures of the tested 20 samples, which was confirmed by mycoplasma-specific gene expression and fluorescence staining. Such contaminated mycoplasma could be successfully eliminated using elimination kit, and proliferation test showed decreased proliferation activity in mycoplasma-contaminated cells. After elimination of contaminated mycoplasma, stem cells from apical papilla showed osteogenic and neural lineage differentiation under certain culture conditions. Our study proposes that the evaluation of mycoplasma contamination and elimination process might be required in the use of stem cells from apical papilla for their potent applications to tissue engineering and regenerative medicine.

  7. Definitions of apical vaginal support loss: a systematic review.

    PubMed

    Meister, Melanie R L; Sutcliffe, Siobhan; Lowder, Jerry L

    2017-03-01

    We sought to identify and summarize definitions of apical support loss utilized for inclusion, success, and failure in surgical trials for treatment of apical vaginal prolapse. Pelvic organ prolapse is a common condition affecting more than 3 million women in the US, and the prevalence is increasing. Prolapse may occur in the anterior compartment, posterior compartment or at the apex. Apical support is considered paramount to overall female pelvic organ support, yet apical support loss is often underrecognized and there are no guidelines for when an apical support procedure should be performed or incorporated into a procedure designed to address prolapse. A systematic literature search was performed in 8 search engines: PubMed 1946-, Embase 1947-, Cochrane Database of Systematic Reviews, Cochrane Database of Abstracts of Review Effects, Cochrane Central Register of Controlled Trials, ClinicalTrials.gov, Proquest Dissertations and Theses, and FirstSearch Proceedings, using key words for apical pelvic organ prolapse and apical suspension procedures through April 2016. Searches were limited to human beings using human filters and articles published in English. Study authors (M.R.L.M., J.L.L.) independently reviewed publications for inclusion based on predefined variables. Articles were eligible for inclusion if they satisfied any of the following criteria: (1) apical support loss was an inclusion criterion in the original study, (2) apical support loss was a surgical indication, or (3) an apical support procedure was performed as part of the primary surgery. A total of 4469 publications were identified. After review, 35 articles were included in the analysis. Prolapse-related inclusion criteria were: (1) apical prolapse (n = 20, 57.1%); (2) overall prolapse (n = 8, 22.8%); or (3) both (n = 6, 17.1%). Definitions of apical prolapse (relative to the hymen) included: (1) apical prolapse >-1 cm (n = 13, 50.0%); (2) apical prolapse >+1 cm (n = 7, 26.9%); (3) apical prolapse >50% of total vaginal length (-[total vaginal length/2]) (n = 4, 15.4%); and (4) cervix/apex >0 cm (n = 2, 7.7%). Sixteen of the 35 studies (45.7%) required the presence of symptoms for inclusion. A measurement of the apical compartment (relative to the hymen) was used as a measure of surgical success or failure in 17 (48.6%) studies. Definitions for surgical success included: (1) prolapse stage >2 in each compartment (n = 5, 29.4%); (2) prolapse >-[total vaginal length/2] (n = 2, 11.8%); (3) apical support >-[total vaginal length/3] (n = 1, 5.9%); (4) absence of prolapse beyond the hymen (n = 1, 5.9%); and (5) point C at ≥-5 cm (n = 2, 11.8%). Surgical failure was defined as: (1) apical prolapse ≥0 cm (n = 2, 11.8%); (2) apical prolapse ≥-1 cm (n = 2, 11.8%); (3) apical prolapse >-[total vaginal length/2] (n = 3, 17.6%); and (4) recurrent apical prolapse surgery (n = 1, 5.9%). Ten (28.6%) of the 35 studies also included symptomatic outcomes in the definition of success or failure. Among randomized, controlled surgical trials designed to address apical vaginal support loss, definitions of clinically significant apical prolapse for study inclusion and surgical success or failure are either highly variable or absent. These findings provide limited evidence of consensus and little insight into current expert opinion. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Diagnosis of apical hypertrophic cardiomyopathy: T-wave inversion and relative but not absolute apical left ventricular hypertrophy☆

    PubMed Central

    Flett, Andrew S.; Maestrini, Viviana; Milliken, Don; Fontana, Mariana; Treibel, Thomas A.; Harb, Rami; Sado, Daniel M.; Quarta, Giovanni; Herrey, Anna; Sneddon, James; Elliott, Perry; McKenna, William; Moon, James C.

    2015-01-01

    Background Diagnosis of apical HCM utilizes conventional wall thickness criteria. The normal left ventricular wall thins towards the apex such that normal values are lower in the apical versus the basal segments. The impact of this on the diagnosis of apical hypertrophic cardiomyopathy has not been evaluated. Methods We performed a retrospective review of 2662 consecutive CMR referrals, of which 75 patients were identified in whom there was abnormal T-wave inversion on ECG and a clinical suspicion of hypertrophic cardiomyopathy. These were retrospectively analyzed for imaging features consistent with cardiomyopathy, specifically: relative apical hypertrophy, left atrial dilatation, scar, apical cavity obliteration or apical aneurysm. For comparison, the same evaluation was performed in 60 healthy volunteers and 50 hypertensive patients. Results Of the 75 patients, 48 met conventional HCM diagnostic criteria and went on to act as another comparator group. Twenty-seven did not meet criteria for HCM and of these 5 had no relative apical hypertrophy and were not analyzed further. The remaining 22 patients had relative apical thickening with an apical:basal wall thickness ratio > 1 and a higher prevalence of features consistent with a cardiomyopathy than in the control groups with 54% having 2 or more of the 4 features. No individual in the healthy volunteer group had more than one feature and no hypertension patient had more than 2. Conclusion A cohort of individuals exist with T wave inversion, relative apical hypertrophy and additional imaging features of HCM suggesting an apical HCM phenotype not captured by existing diagnostic criteria. PMID:25666123

  9. Apical stress distribution under vertical compaction of gutta-percha and occlusal loads in canals with varying apical sizes: a three-dimensional finite element analysis.

    PubMed

    Yuan, K; Niu, C; Xie, Q; Jiang, W; Gao, L; Ma, R; Huang, Z

    2018-02-01

    To investigate and compare the effects of two apical canal instrumentation protocols on apical stress distribution at the root apex under vertical compaction of gutta-percha and occlusal loads using finite element analysis. Three finite element analysis models of a mandibular first premolar were reconstructed: an original canal model, a size 35, .04 taper apical canal enlargement model and a Lightspeed size 60 apical canal enlargement model. A 15 N compaction force was applied vertically to the gutta-percha 5 mm from the apex. A 175 N occlusal load in two directions (vertical and 45° to the longitudinal axis of the tooth) was simulated. Stresses in the apical 2 mm of the root were calculated and compared among the three models. Under vertical compaction, stresses in the apical canal instrumented by Lightspeed size 60 (maximal 3.3 MPa) were higher than that of the size 35, .04 taper model (maximal 1.3 MPa). In the case of the two occlusal forces, the Lightspeed size 60 apical enlargement was associated with the greatest stress distribution in the apical region. The greatest stress and the most obvious stress difference between the models appeared at the tip of the root when occlusal and vertical compaction loads were applied. Apical enlargement caused stress distribution changes in the apical region of roots. The larger apical size led to higher stress concentration at the root apex. © 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  10. Integration of light and metabolic signals for stem cell activation at the shoot apical meristem

    PubMed Central

    Pfeiffer, Anne; Janocha, Denis; Dong, Yihan; Medzihradszky, Anna; Schöne, Stefanie; Daum, Gabor; Suzaki, Takuya; Forner, Joachim; Langenecker, Tobias; Rempel, Eugen; Schmid, Markus; Wirtz, Markus; Hell, Rüdiger; Lohmann, Jan U

    2016-01-01

    A major feature of embryogenesis is the specification of stem cell systems, but in contrast to the situation in most animals, plant stem cells remain quiescent until the postembryonic phase of development. Here, we dissect how light and metabolic signals are integrated to overcome stem cell dormancy at the shoot apical meristem. We show on the one hand that light is able to activate expression of the stem cell inducer WUSCHEL independently of photosynthesis and that this likely involves inter-regional cytokinin signaling. Metabolic signals, on the other hand, are transduced to the meristem through activation of the TARGET OF RAPAMYCIN (TOR) kinase. Surprisingly, TOR is also required for light signal dependent stem cell activation. Thus, the TOR kinase acts as a central integrator of light and metabolic signals and a key regulator of stem cell activation at the shoot apex. DOI: http://dx.doi.org/10.7554/eLife.17023.001 PMID:27400267

  11. YAP and TAZ in epithelial stem cells: A sensor for cell polarity, mechanical forces and tissue damage.

    PubMed

    Elbediwy, Ahmed; Vincent-Mistiaen, Zoé I; Thompson, Barry J

    2016-07-01

    The YAP/TAZ family of transcriptional co-activators drives cell proliferation in epithelial tissues and cancers. Yet, how YAP and TAZ are physiologically regulated remains unclear. Here we review recent reports that YAP and TAZ act primarily as sensors of epithelial cell polarity, being inhibited when cells differentiate an apical membrane domain, and being activated when cells contact the extracellular matrix via their basal membrane domain. Apical signalling occurs via the canonical Crumbs/CRB-Hippo/MST-Warts/LATS kinase cascade to phosphorylate and inhibit YAP/TAZ. Basal signalling occurs via Integrins and Src family kinases to phosphorylate and activate YAP/TAZ. Thus, YAP/TAZ is localised to the nucleus in basal stem/progenitor cells and cytoplasm in differentiated squamous cells or columnar cells. In addition, other signals such as mechanical forces, tissue damage and possibly receptor tyrosine kinases (RTKs) can influence MST-LATS or Src family kinase activity to modulate YAP/TAZ activity. © 2016 The Authors BioEssays Published by WILEY Periodicals, Inc.

  12. [Ultrastructure and cytochemistry of the pellicle and apical complexes of the kinete of Babesia bigemina and Babesia ovis in the hemolymph and oavry of the tick].

    PubMed

    Weber, G

    1980-02-01

    The term kinete is used in this paper for the cigar-shaped, motile development stages (VERMICULE") OF Babesia occurring intra- and extracellularly in hemolymph and overy (including oocytes) of vectors, hard ticks (Ixodoidea). The structure of, and cytochemical activities of hydrolases (acid phosphatase, nonspecific esterase) in the pellicle and the apical complex was studied at the fine-structural level in kinetes of Babesia bigemina Smith & Kilborne, in hemolympho of female Boophilus microplus Canestrini. The cytochemistry of acid hydrolases was studied also in kinetes of Babesia ovis (Babès) Starcovici, in hemolymph and ovary of Rhipicephalus bursa Canestrini & Fanzago. The pellicle of the B. bigemina kinetes is composted of 3 membranes (pellicular complex): an outer membrane, approximately 8 nm thick (the plasmalemma) and 2 innder ones, each approximately nm thick, lying closely together. The outer membrane appears to be covered by a structureless coat, 3 nm thick. The space between the inner double membrane and the plasmalemma is 7.5 nm. The whole pellicular complex is 30 nm in diameter. The 2 inner pellicular membranes appear to be derived from the endoplasmic reticulum (ER) for the following reasons: (a) a layer of hydrolase-active material is enclosed by these membranes; (b) in the spheroid parasite stages which transform from kinetes inside hemocytes, the inner double membrane is apparently replaced by an ER cisterna; (c) the thickness of each of the inner pellicular membranes is approximately the same as that of the ER membrane. There are circular openings in the pellicular double membrane with average diameters of 100 nm; despite some similarity to micropores, they have a specific structure. The term Intrapellikularfenster (IPF) (intrapellicular windows) or pseudomicropores is proposed for these pellicular differentiations. The margin of an IPF is formed by the 2 inner membranes folding into each other; cytoplasmic, electron-dense material is accumulated alongside this edge. Unlike that of micropores, the plasmalemma of the IPF is not invaginated. The IPF appears as a single, dark ring in tangential sections. At times, rhoptry-like bodies are associated with the openings. The function of the IPF is not known. An intrapellicular opening similar to the IPF, although wider, is present at the apex of the parasite. Its margin coincides with the inners edge of the apical ring. Typical subpellicular microtubuli were not observed in the Babesia kinetes. The apical complex of the B. bigemina kinetes consists of an Apikalschirm (apical umbrella), a crown of microtubuli beneath it, and rhoptries: micronemes are also present in large numbers. The Apikalschirm is located beneath the pellicle of the apical pole of the parasite. It is a wheel-like structure composed of spokes radiating from a wide, hub=like central ring (apical ring). It should be stressed that the apical ring is not identical with the polar ring described as an integral part of the pellicular complex in other Apicomplexa...

  13. Does apical root resection in endodontic microsurgery jeopardize the prosthodontic prognosis?

    PubMed Central

    Cho, Sin-Yeon

    2013-01-01

    Apical surgery cuts off the apical root and the crown-to-root ratio becomes unfavorable. Crown-to-root ratio has been applied to periodontally compromised teeth. Apical root resection is a different matter from periodontal bone loss. The purpose of this paper is to review the validity of crown-to-root ratio in the apically resected teeth. Most roots have conical shape and the root surface area of coronal part is wider than apical part of the same length. Therefore loss of alveolar bone support from apical resection is much less than its linear length.The maximum stress from mastication concentrates on the cervical area and the minimum stress was found on the apical 1/3 area. Therefore apical root resection is not so harmful as periodontal bone loss. Osteotomy for apical resection reduces longitudinal width of the buccal bone and increases the risk of endo-perio communication which leads to failure. Endodontic microsurgery is able to realize 0 degree or shallow bevel and precise length of root resection, and minimize the longitudinal width of osteotomy. The crown-to-root ratio is not valid in evaluating the prosthodontic prognosis of the apically resected teeth. Accurate execution of endodontic microsurgery to preserve the buccal bone is essential to avoid endo-perio communication. PMID:23741707

  14. Apical sorting of lysoGPI-anchored proteins occurs independent of association with detergent-resistant membranes but dependent on their N-glycosylation.

    PubMed

    Castillon, Guillaume Alain; Michon, Laetitia; Watanabe, Reika

    2013-06-01

    Most glycosylphosphatidylinositol-anchored proteins (GPI-APs) are located at the apical surface of epithelial cells. The apical delivery of GPI-APs is believed to result from their association with lipid rafts. We find that overexpression of C-terminally tagged PGAP3 caused predominant production of lysoGPI-APs, an intermediate precursor in the GPI lipid remodeling process in Madin-Darby canine kidney cells. In these cells, produced lysoGPI-APs are not incorporated into detergent-resistant membranes (DRMs) but still are delivered apically, suggesting that GPI-AP association with DRMs is not necessary for apical targeting. In contrast, apical transport of both fully remodeled and lyso forms of GPI-APs is dependent on N-glycosylation, confirming a general role of N-glycans in apical protein transport. We also find that depletion of cholesterol causes apical-to-basolateral retargeting not only of fully remodeled GPI-APs, but also of lysoGPI-APs, as well as endogenous soluble and transmembrane proteins that would normally be targeted to the apical membrane. These findings confirm the essential role for cholesterol in the apical protein targeting and further demonstrate that the mechanism of cholesterol-dependent apical sorting is not related to DRM association of GPI-APs.

  15. Drought and Heat Differentially Affect XTH Expression and XET Activity and Action in 3-Day-Old Seedlings of Durum Wheat Cultivars with Different Stress Susceptibility

    PubMed Central

    Iurlaro, Andrea; De Caroli, Monica; Sabella, Erika; De Pascali, Mariarosaria; Rampino, Patrizia; De Bellis, Luigi; Perrotta, Carla; Dalessandro, Giuseppe; Piro, Gabriella; Fry, Stephen C.; Lenucci, Marcello S.

    2016-01-01

    Heat and drought stress have emerged as major constraints for durum wheat production. In the Mediterranean area, their negative effect on crop productivity is expected to be exacerbated by the occurring climate change. Xyloglucan endotransglucosylase/hydrolases (XTHs) are chief enzymes in cell wall remodeling, whose relevance in cell expansion and morphogenesis suggests a central role in stress responses. In this work the potential role of XTHs in abiotic stress tolerance was investigated in durum wheat. The separate effects of dehydration and heat exposure on XTH expression and its endotransglucosylase (XET) in vitro activity and in vivo action have been monitored, up to 24 h, in the apical and sub-apical root regions and shoots excised from 3-day-old seedlings of durum wheat cultivars differing in stress susceptibility/tolerance. Dehydration and heat stress differentially influence the XTH expression profiles and the activity and action of XET in the wheat seedlings, depending on the degree of susceptibility/tolerance of the cultivars, the organ, the topological region of the root and, within the root, on the gradient of cell differentiation. The root apical region was the zone mainly affected by both treatments in all assayed cultivars, while no change in XET activity was observed at shoot level, irrespective of susceptibility/tolerance, confirming the pivotal role of the root in stress perception, signaling, and response. Conflicting effects were observed depending on stress type: dehydration evoked an overall increase, at least in the apical region of the root, of XET activity and action, while a significant inhibition was caused by heat treatment in most cultivars. The data suggest that differential changes in XET action in defined portions of the root of young durum wheat seedlings may have a role as a response to drought and heat stress, thus contributing to seedling survival and crop establishment. A thorough understanding of the mechanisms underlying these variations could represent the theoretical basis for implementing breeding strategies to develop new highly productive hybrids adapted to future climate scenarios. PMID:27891140

  16. Spatial Segregation and Interaction of Calcium Signalling Mechanisms in Rat Hippocampal CA1 Pyramidal Neurons

    PubMed Central

    Nakamura, Takeshi; Lasser-Ross, Nechama; Nakamura, Kyoko; Ross, William N

    2002-01-01

    Postsynaptic [Ca2+]i increases result from Ca2+ entry through ligand-gated channels, entry through voltage-gated channels, or release from intracellular stores. We found that these sources have distinct spatial distributions in hippocampal CA1 pyramidal neurons. Large amplitude regenerative release of Ca2+ from IP3-sensitive stores in the form of Ca2+ waves were found almost exclusively on the thick apical shaft. Smaller release events did not extend more than 15 μm into the oblique dendrites. These synaptically activated regenerative waves initiated at points where the stimulated oblique dendrites branch from the apical shaft. In contrast, NMDA receptor-mediated increases were observed predominantly in oblique dendrites where spines are found at high density. These [Ca2+]i increases were typically more than eight times larger than [Ca2+]i from this source on the main aspiny apical shaft. Ca2+ entry through voltage-gated channels, activated by backpropagating action potentials, was detected at all dendritic locations. These mechanisms were not independent. Ca2+ entry through NMDA receptor channels or voltage-gated channels (as previously demonstrated) synergistically enhanced Ca2+ release generated by mGluR mobilization of IP3. PMID:12205182

  17. Cdk1 Activates Pre-mitotic Nuclear Envelope Dynein Recruitment and Apical Nuclear Migration in Neural Stem Cells.

    PubMed

    Baffet, Alexandre D; Hu, Daniel J; Vallee, Richard B

    2015-06-22

    Dynein recruitment to the nuclear envelope is required for pre-mitotic nucleus-centrosome interactions in nonneuronal cells and for apical nuclear migration in neural stem cells. In each case, dynein is recruited to the nuclear envelope (NE) specifically during G2 via two nuclear pore-mediated mechanisms involving RanBP2-BicD2 and Nup133-CENP-F. The mechanisms responsible for cell-cycle control of this behavior are unknown. We now find that Cdk1 serves as a direct master controller for NE dynein recruitment in neural stem cells and HeLa cells. Cdk1 phosphorylates conserved sites within RanBP2 and activates BicD2 binding and early dynein recruitment. Late recruitment is triggered by a Cdk1-induced export of CENP-F from the nucleus. Forced NE targeting of BicD2 overrides Cdk1 inhibition, fully rescuing dynein recruitment and nuclear migration in neural stem cells. These results reveal how NE dynein recruitment is cell-cycle regulated and identify the trigger mechanism for apical nuclear migration in the brain. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Apical ammonium inhibition of cAMP-stimulated secretion in T84 cells is bicarbonate dependent.

    PubMed

    Worrell, Roger T; Best, Alison; Crawford, Oscar R; Xu, Jie; Soleimani, Manoocher; Matthews, Jeffrey B

    2005-10-01

    Normal human colonic luminal (NH(4)(+)) concentration ([NH(4)(+)]) ranges from approximately 10 to 100 mM. However, the nature of the effects of NH(4)(+) on transport, as well as NH(4)(+) transport itself, in colonic epithelium is poorly understood. We elucidate here the effects of apical NH(4)(+) on cAMP-stimulated Cl(-) secretion in colonic T84 cells. In HEPES-buffered solutions, 10 mM apical NH(4)(+) had no significant effect on cAMP-stimulated current. In contrast, 10 mM apical NH(4)(+) reduced current within 5 min to 61 +/- 4% in the presence of 25 mM HCO(3)(-). Current inhibition was not simply due to an increase in extracellular K(+)-like cations, in that the current magnitude was 95 +/- 5% with 10 mM apical K(+) and 46 +/- 3% with 10 mM apical NH(4)(+) relative to that with 5 mM apical K(+). We previously demonstrated that inhibition of Cl(-) secretion by basolateral NH(4)(+) occurs in HCO(3)(-)-free conditions and exhibits anomalous mole fraction behavior. In contrast, apical NH(4)(+) inhibition of current in HCO(3)(-) buffer did not show anomalous mole fraction behavior and followed the absolute [NH(4)(+)] in K(+)-NH(4)(+) mixtures, where K(+) concentration + [NH(4)(+)] = 10 mM. The apical NH(4)(+) inhibitory effect was not prevented by 100 microM methazolamide, suggesting no role for apical carbonic anhydrase. However, apical NH(4)(+) inhibition of current was prevented by 10 min of pretreatment of the apical surface with 500 microM DIDS, 100 microM 4,4'-dinitrostilbene-2,2'-disulfonic acid (DNDS), or 25 microM niflumic acid, suggesting a role for NH(4)(+) action through an apical anion exchanger. mRNA and protein for the apical anion exchangers SLC26A3 [downregulated in adenoma (DRA)] and SLC26A6 [putative anion transporter (PAT1)] were detected in T84 cells by RT-PCR and Northern and Western blots. DRA and PAT1 appear to associate with CFTR in the apical membrane. We conclude that the HCO(3)(-) dependence of apical NH(4)(+) inhibition of secretion is due to the action of NH(4)(+) on an apical anion exchanger.

  19. Salicylic Acid Regulates Pollen Tip Growth through an NPR3/NPR4-Independent Pathway.

    PubMed

    Rong, Duoyan; Luo, Nan; Mollet, Jean Claude; Liu, Xuanming; Yang, Zhenbiao

    2016-11-07

    Tip growth is a common strategy for the rapid elongation of cells to forage the environment and/or to target to long-distance destinations. In the model tip growth system of Arabidopsis pollen tubes, several small-molecule hormones regulate their elongation, but how these rapidly diffusing molecules control extremely localized growth remains mysterious. Here we show that the interconvertible salicylic acid (SA) and methylated SA (MeSA), well characterized for their roles in plant defense, oppositely regulate Arabidopsis pollen tip growth with SA being inhibitory and MeSA stimulatory. The effect of SA and MeSA was independent of known NPR3/NPR4 SA receptor-mediated signaling pathways. SA inhibited clathrin-mediated endocytosis in pollen tubes associated with an increased accumulation of less stretchable demethylated pectin in the apical wall, whereas MeSA did the opposite. Furthermore, SA and MeSA alter the apical activation of ROP1 GTPase, a key regulator of tip growth in pollen tubes, in an opposite manner. Interestingly, both MeSA methylesterase and SA methyltransferase, which catalyze the interconversion between SA and MeSA, are localized at the apical region of pollen tubes, indicating of the tip-localized production of SA and MeSA and consistent with their effects on the apical cellular activities. These findings suggest that local generation of a highly diffusible signal can regulate polarized cell growth, providing a novel mechanism of cell polarity control apart from the one involving protein and mRNA polarization. Copyright © 2016 The Author. Published by Elsevier Inc. All rights reserved.

  20. Exogenous retinoic acid induces digit reduction in opossums (Monodelphis domestica) by disrupting cell death and proliferation, and apical ectodermal ridge and zone of polarizing activity function.

    PubMed

    Molineaux, Anna C; Maier, Jennifer A; Schecker, Teresa; Sears, Karen E

    2015-03-01

    Retinoic acid (RA) is a vitamin A derivative. Exposure to exogenous RA generates congenital limb malformations (CLMs) in species from frogs to humans. These CLMs include but are not limited to oligodactyly and long-bone hypoplasia. The processes by which exogenous RA induces CLMs in mammals have been best studied in mouse, but as of yet remain unresolved. We investigated the impact of exogenous RA on the cellular and molecular development of the limbs of a nonrodent model mammal, the opossum Monodelphis domestica. Opossums exposed to exogenous retinoic acid display CLMs including oligodactly, and results are consistent with opossum development being more susceptible to RA-induced disruptions than mouse development. Exposure of developing opossums to exogenous RA leads to an increase in cell death in the limb mesenchyme that is most pronounced in the zone of polarizing activity, and a reduction in cell proliferation throughout the limb mesenchyme. Exogenous RA also disrupts the expression of Shh in the zone of polarizing activity, and Fgf8 in the apical ectodermal ridge, and other genes with roles in the regulation of limb development and cell death. Results are consistent with RA inducing CLMs in opossum limbs by disrupting the functions of the apical ectodermal ridge and zone of polarizing activity, and driving an increase in cell death and reduction of cell proliferation in the mesenchyme of the developing limb. © 2015 Wiley Periodicals, Inc.

  1. Efficacy of four different irrigation techniques combined with 60 °C 3% sodium hypochlorite and 17% EDTA in smear layer removal.

    PubMed

    Guo, Xiangjun; Miao, Hui; Li, Lei; Zhang, Shasha; Zhou, Dongyan; Lu, Yan; Wu, Ligeng

    2014-09-08

    Efforts to improve the efficacy of smear layer removal by applying irrigant activation at the final irrigation or by elevating the temperature of the irrigant have been reported. However, the combination of such activation protocols with 60 °C 3% sodium hypochlorite (NaOCl) has seldom been mentioned. The aim of this study was to compare the efficacy in smear layer removal of four different irrigation techniques combined with 60 °C 3% NaOCl and 17% EDTA. Fifty single-rooted teeth were randomly divided into five groups (n = 10) according to the irrigant agitation protocols used during chemomechanical preparation(Dentsply Maillefer, Ballaigues, Switzerland): a side-vented needle group, a ultrasonic irrigation (UI) group, a NaviTip FX group, an EndoActivator group, and a control group (no agitation). After each instrumentation, the root canals were irrigated with 1 mL of 3% NaOCl at 60 °C for 1 minute, and after the whole instrumentation, the root canals were rinsed with 1 mL of 17% EDTA for 1 minute. Both NaOCl and EDTA were activated with one of the five irrigation protocols. The efficacy of smear layer removal was scored at the apical, middle and coronal thirds. The Data were statistically analyzed using SAS version 9.2 for Windows (rank sum test for a randomised block design and ANOVA). No significant differences among the NaviTip FX group, EndoActivator group and control groups, and each of these groups showed a lower score than that of UI group (P < 0.05). Within each group, all three thirds were ranked in the following order: coronal > middle > apical (P < 0.05). In the coronal third, the NaviTip FX group was better than UI group. In the middle and apical third, the differences were not significant among any of the groups. Even without any activation, the combination of 60 °C 3% NaOCl and 17% EDTA could remove the smear layer effectively, similar to NaviTip FX or EndoActivator, and these three protocols were more effective than UI. However, regardless of different types of irrigation technique applied, complete removal of the smear layer was not achieved, particularly in the apical third.

  2. Neisseria gonorrhoeae infects the human endocervix by activating non-muscle myosin II-mediated epithelial exfoliation

    PubMed Central

    Yu, Qian; Lin, Brian; Qiu, Jessica; Stein, Daniel C.

    2017-01-01

    Colonization and disruption of the epithelium is a major infection mechanism of mucosal pathogens. The epithelium counteracts infection by exfoliating damaged cells while maintaining the mucosal barrier function. The sexually transmitted bacterium Neisseria gonorrhoeae (GC) infects the female reproductive tract primarily from the endocervix, causing gonorrhea. However, the mechanism by which GC overcome the mucosal barrier remains elusive. Using a new human tissue model, we demonstrate that GC can penetrate into the human endocervix by inducing the exfoliation of columnar epithelial cells. We found that GC colonization causes endocervical epithelial cells to shed. The shedding results from the disassembly of the apical junctions that seal the epithelial barrier. Apical junction disruption and epithelial exfoliation increase GC penetration into the endocervical epithelium without reducing bacterial adherence to and invasion into epithelial cells. Both epithelial exfoliation and junction disruption require the activation and accumulation of non-muscle myosin II (NMII) at the apical surface and GC adherent sites. GC inoculation activates NMII by elevating the levels of the cytoplasmic Ca2+ and NMII regulatory light chain phosphorylation. Piliation of GC promotes, but the expression of a GC opacity-associated protein variant, OpaH that binds to the host surface proteins CEACAMs, inhibits GC-induced NMII activation and reorganization and Ca2+ flux. The inhibitory effects of OpaH lead to reductions in junction disruption, epithelial exfoliation, and GC penetration. Therefore, GC phase variation can modulate infection in the human endocervix by manipulating the activity of NMII and epithelial exfoliation. PMID:28406994

  3. Transcriptome Analysis of Tomato Flower Pedicel Tissues Reveals Abscission Zone-Specific Modulation of Key Meristem Activity Genes

    PubMed Central

    Sun, Xiuli; Zhang, Rongzhi; Wu, Liang; Liang, Yanchun; Mao, Long

    2013-01-01

    Tomato flower abscises at the anatomically distinct abscission zone that separates the pedicel into basal and apical portions. During abscission, cell separation occurs only at the abscission zone indicating distinctive molecular regulation in its cells. We conducted a transcriptome analysis of tomato pedicel tissues during ethylene promoted abscission. We found that the abscission zone was the most active site with the largest set of differentially expressed genes when compared with basal and apical portions. Gene Ontology analyses revealed enriched transcription regulation and hydrolase activities in the abscission zone. We also demonstrate coordinated responses of hormone and cell wall related genes. Besides, a number of ESTs representing homologs of key Arabidopsis shoot apical meristem activity genes were found to be preferentially expressed in the abscission zone, including WUSCHEL (WUS), KNAT6, LATERAL ORGAN BOUNDARIES DOMAIN PROTEIN 1(LBD1), and BELL-like homeodomain protein 1 (BLH1), as well as tomato axillary meristem genes BLIND (Bl) and LATERAL SUPPRESSOR (Ls). More interestingly, the homologs of WUS and the potential functional partner OVATE FAMILIY PROTEIN (OFP) were subsequently down regulated during abscission while Bl and AGL12 were continuously and specifically induced in the abscission zone. The expression patterns of meristem activity genes corroborate the idea that cells of the abscission zone confer meristem-like nature and coincide with the course of abscission and post-abscission cell differentiation. Our data therefore propose a possible regulatory scheme in tomato involving meristem genes that may be required not only for the abscission zone development, but also for abscission. PMID:23390523

  4. A unique approach to demonstrating that apical bud temperature specifically determines leaf initiation rate in the dicot Cucumis sativus.

    PubMed

    Savvides, Andreas; Dieleman, Janneke A; van Ieperen, Wim; Marcelis, Leo F M

    2016-04-01

    Leaf initiation rate is largely determined by the apical bud temperature even when apical bud temperature largely deviates from the temperature of other plant organs. We have long known that the rate of leaf initiation (LIR) is highly sensitive to temperature, but previous studies in dicots have not rigorously demonstrated that apical bud temperature controls LIR independent of other plant organs temperature. Many models assume that apical bud and leaf temperature are the same. In some environments, the temperature of the apical bud, where leaf initiation occurs, may differ by several degrees Celsius from the temperature of other plant organs. In a 28-days study, we maintained temperature differences between the apical bud and the rest of the individual Cucumis sativus plants from -7 to +8 °C by enclosing the apical buds in transparent, temperature-controlled, flow-through, spheres. Our results demonstrate that LIR was completely determined by apical bud temperature independent of other plant organs temperature. These results emphasize the need to measure or model apical bud temperatures in dicots to improve the prediction of crop development rates in simulation models.

  5. The Incremental Value of Magnetic Resonance Imaging for Identification of Apical Pouch in Patients with Apical Variant of Hypertrophic Cardiomyopathy.

    PubMed

    Vucicevic, Darko; Lester, Steven J; Appleton, Christopher P; Panse, Prasad M; Schleifer, John William; Wilansky, Susan

    2016-04-01

    The development of a left ventricular (LV) apical pouch in patients with apical hypertrophic cardiomyopathy (aHCM) has been thought to be the transition point that can become an apical aneurysm, which is linked to higher risk of adverse events. In our study, we sought to compare the ability of transthoracic echocardiography (echo) and cardiac magnetic resonance imaging (cMRI) to accurately identify the presence of an apical pouch or aneurysm in patients with aHCM. We retrospectively reviewed the charts of all consecutive patients that had features of aHCM on imaging. Data from cMRI and echo examinations were abstracted, and the ability of these diagnostic modalities to identify the presence of a LV apical pouch and aneurysm was analyzed. Of 31 patients with aHCM, 17 (54.8%) had an apical pouch and 2 were found to have apical aneurysm (6.5%) on cMRI. Echo with and without perflutren contrast was able to accurately identify both aneurysms, but only 47.1% (8/17) of apical pouches seen by cMRI. Two patients had apical thrombus that was identified by cMRI, but not by echo. Our findings indicate that cMRI is superior to echo in identifying apical pouches in patients with aHCM. Our results also suggest that in patients undergoing echo, the use of perflutren contrast for LV opacification increases the diagnostic yield. Further study is necessary to delineate whether earlier identification of an apical pouch will be of clinical benefit for patients with aHCM by altering clinical management and avoiding adverse cardiovascular events. © 2015, Wiley Periodicals, Inc.

  6. Trends in Hysteropexy and Apical Support for Uterovaginal Prolapse in the United States from 2002 to 2012.

    PubMed

    Madsen, Annetta M; Raker, Christina; Sung, Vivian W

    Our objective was to describe trends in hysteropexy and apical support for uterovaginal prolapse (UVP) from 2002 to 2012 in the United States. We identified patient and hospital variables associated with hysteropexy and apical support. We used the Nationwide Inpatient Sample and International Classification of Diseases, Ninth Revision codes to identify a population of women 18 years or older with UVP undergoing pelvic organ prolapse surgery from January 1, 2002, to December 31, 2012. Procedures were categorized as (1) hysteropexy, (2) obliterative with uterine preservation, (3) hysterectomy with apical support, (4) hysterectomy without apical support, and (5) other reconstruction without apical support. Categories were dichotomized into those with and without apical support. We used survey weights to obtain nationally representative estimates; χ and linear and logistic regression compared procedure groups. An estimated 815,184 hospital discharges of pelvic organ prolapse procedures for UVP occurred from 2002 to 2012. During this time, hysteropexies increased from 1.81% to 5.00% (P < 0.0001). From 2002 to 2012, hysterectomies with apical support increased (10.07% to 32.51%, P < 0.0001), hysterectomy without apical support decreased (27.14% to 17.12%, P < 0.0001), and reconstruction without apical support decreased (59.07% to 40.48%, P < 0.0001). In most recent years 2011 to 2012, 60% of women with UVP underwent inpatient surgery without an apical procedure. Age 52 years or older, Medicare payment, Northeast region, and urban teaching hospitals were associated with increased odds of apical support for UVP (P < 0.001 for all). Hysteropexy significantly increased in the United States from 2002 to 2012, although the overall proportion remains low. While hysterectomy without apical support is decreasing, approximately 60% of inpatient procedures performed for UVP do not address the apex.

  7. Bone physiology in human grafted and non-grafted extraction sockets--an immunohistochemical study.

    PubMed

    Nahles, Susanne; Nack, Claudia; Gratecap, Kerrin; Lage, Hermann; Nelson, John J; Nelson, Katja

    2013-07-01

    The aim of the present immunohistological investigation was to define and compare the osteogenic potential with the vascularization of the provisional matrix in grafted and ungrafted extraction sockets after 4 and 12 weeks of healing. A total of 33 Patients (15 women, 18 men) with 65 extraction sites with a mean age of 54.4 years (30-73 years) participated in this study. After tooth extraction, the sockets were augmented with Bio-Oss collagen or non-augmented. At implant placement after 4 or 12 weeks bone biopsies were obtained. Within the specimens the osteogenic and endothelial potential of mesenchymal cells was analyzed in the provisional matrix using immunohistochemical analysis with three monoclonal antibodies Cbfa1/Runx2, Osteocalcin (OC), and CD31. Statistical analysis was performed using Mann-Whitney U-test, Spearman's rank-order correlation coefficient, and the two-factorial analysis for repeated measurements. Of the 65 extraction sockets, 25 (13 non-augmented, 12 augmented) sites after 4 weeks healing time and 40 (19 non-augmented, 21 augmented) sites after 12 weeks healing time were involved in the study. No signs of acute or chronic inflammation were noted in any specimens. After 4 weeks, a median amount of 56% (10-85%) of Cbfa1 positive cells and a median amount of cells expressing OC of 21% (5-42%) were measured. A median CD31 score of 5 was observed. After 12 weeks, a median amount of 61% (19-90%) positive cells expressed by Cbfa1/Runx2 staining a median amount of OC positive cells of 9% (2-17%) was measured. The results at 12 weeks revealed a median score of CD31 positive cells of 3. Osteoblastic activity in the provisional matrix was highest after 4 weeks of healing period. The active zone of bone formation is found in the apical region of the extraction socket during the early healing phase, shifting to the coronal region after 12 weeks. A peak of osteoblast activity within the first weeks is followed by a reduction in mature osteoblasts with osteoblasts remaining in an inactive stage. The vascularity changed in likewise fashion to the maturation of osteoblasts within the observation period. The results have shown that with increasing age a decreasing endothelial potential was observed not after 4 weeks, but after 12 weeks, thus it suggests that angiogenesis is diminished in older patients in the later phase of healing in extraction sockets. © 2012 John Wiley & Sons A/S.

  8. Apical Extrusion of Irrigants in Immature Permanent Teeth by Using EndoVac and Needle Irrigation: An In Vitro Study

    PubMed Central

    Velmurugan, N; Sooriaprakas, C; Jain, Preetham

    2014-01-01

    Objective: Immature teeth have a large apical opening and thin divergent or parallel dentinal walls; hence, with conventional needle irrigation there is a very high possibility of extrusion. This study was done to compare the apical extrusion of NaOCl in an immature root delivered using EndoVac and needle irrigation. Materials and Methods: Eighty freshly extracted maxillary central incisors were decoronated followed by access cavity preparation. Modified organotypic protocol was performed to create an open apex; then, the samples were divided into four groups (n=20): EndoVac Microcannula (group I), EndoVac Macrocannula (group II), NaviTip irrigation needle (group III) and Max-i-Probe Irrigating needle (group IV); 9.0 ml of 3% sodium hypochlorite was delivered slowly over a period of 60 seconds. Extruded irrigants were collected in a vial and analysed statistically. Results: Group I, group III and group IV showed 100% extrusion (20/20) but group II showed only 40% extrusion (8/20). The difference in this respect between group II and other groups was statistically significant (P<0.001). With regards to the volume of extrusion, group II had only 0.23 ml of extruded irrigant. Group I extruded 7.53ml of the irrigant. Group III and group IV extruded the entire volume of irrigant delivered. Conclusion: EndoVac Macrocannula resulted in the least extrusion of irrigant in immature teeth when compared to EndoVac Microcannula and conventional needle irrigation. PMID:25584055

  9. Comparison of apical transportation between ProFile and ProTaper NiTi rotary instruments.

    PubMed

    Iqbal, M K; Firic, S; Tulcan, J; Karabucak, B; Kim, S

    2004-06-01

    To use a newly developed radiographic technique to compare apical transportation and loss of working length (WL) between.06 taper ProFile Series 29 and ProTaper nickel-titanium (NiTi) rotary instruments in vitro. Mesio-buccal canals of 40 extracted mandibular molars were randomly divided into two groups. Group 1 was instrumented with ProFile and group 2 with ProTaper instruments according to the manufacturers' directions. A specially constructed radiographic jig with a Schick digital radiographic system (Schick Technologies Inc., Long Island City, NY, USA) was used to take pre- and postoperative radiographs of the samples at predetermined angulations. Using AutoCAD 2000 (Autodesk Inc., San Rafael, CA, USA), the central axes of initial and final instruments were radiographically superimposed to determine the loss of WL and degree of transportation at D(0), D(1), D(2) and D(4) from the WL. Data were analysed using repeated-measures anova. A statistically significant difference in apical transportation was found at the D(4) level between the two groups (P = 0.05). There was no statistical significance regarding postinstrumentation change in WL between groups. Spearman's Bivariate Correlation analysis indicated no statistically significant relationship between the radius of curvature and transportation. The results indicate that both ProTaper and ProFile instruments are comparable to each other in regards to their ability to optimally enlarge root canal with minimal transportation and loss of WL in vitro.

  10. Semiconductor laser irradiation improves root canal sealing during routine root canal therapy

    PubMed Central

    Hu, Xingxue; Wang, Dashan; Cui, Ting; Yao, Ruyong

    2017-01-01

    Objective To evaluate the effect of semiconductor laser irradiation on root canal sealing after routine root canal therapy (RCT). Methods Sixty freshly extracted single-rooted human teeth were randomly divided into six groups (n = 10). The anatomic crowns were sectioned at the cementoenamel junction and the remaining roots were prepared endodontically with conventional RCT methods. Groups A and B were irradiated with semiconductor laser at 1W for 20 seconds; Groups C and D were ultrasonically rinsed for 60 seconds as positive control groups; Groups E and F without treatment of root canal prior to RCT as negative control groups. Root canal sealing of Groups A, C and E were evaluated by measurements of apical microleakage. The teeth from Groups B, D and F were sectioned, and the micro-structures were examined with scanning electron microscopy (SEM). One way ANOVA and LSD-t test were used for statistical analysis (α = .05). Results The apical sealing of both the laser irradiated group and the ultrasonic irrigated group were significantly different from the control group (p<0.5). There was no significant difference between the laser irradiated group and the ultrasonic irrigated group (p>0.5). SEM observation showed that most of the dentinal tubules in the laser irradiation group melted, narrowed or closed, while most of the dentinal tubules in the ultrasonic irrigation group were filled with tooth paste. Conclusion The application of semiconductor laser prior to root canal obturation increases the apical sealing of the roots treated. PMID:28957407

  11. Chemomechanical preparation by hand instrumentation and by Mtwo engine-driven rotary files, an ex vivo study.

    PubMed

    Krajczár, Károly; Tigyi, Zoltán; Papp, Viktória; Marada, Gyula; Sára, Jeges; Tóth, Vilmos

    2012-07-01

    To compare the disinfecting efficacy of the sodium hypochlorite irrigation by root canal preparation with stainless steel hand files, taper 0.02 and nickel-titanium Mtwo files with taper 0.04-0.06. 40 extracted human teeth were sterilized, and then inoculated with Enterococcus faecalis (ATCC 29212). After 6 day incubation time the root canals were prepared by hand with K-files (n=20) and by engine-driven Mtwo files (VDW, Munich, Germany) (n=20). Irrigation was carried out with 2.5% NaOCl in both cases. Samples were taken and determined in colony forming units (CFU) from the root canals before and after the preparation with instruments #25 and #35. Significant reduction in bacterial count was determined after filing at both groups. The number of bacteria kept on decreasing with the extension of apical preparation diameter. There was no significant difference between the preparation sizes in the bacterial counts after hand or engine-driven instrumentation at the same apical size. Statistical analysis was carried out with Mann-Whitney test, paired t-test and independent sample t-test. Significant reduction in CFU was achieved after the root canal preparation completed with 2.5% NaOCl irrigation, both with stainless steel hand or nickel-titanium rotary files. The root canal remained slightly infected after chemo mechanical preparation in both groups. Key words:Chemomechanical preparation, root canal disinfection, nickel-titanium, conicity, greater taper, apical size.

  12. Evaluation of Root Canal Cleaning and Shaping Efficacy of Three Engine-driven Instruments: SafeSider, ProTaper Universal and Lightspeed LSX.

    PubMed

    Wigler, Ronald; Koren, Tal; Tsesis, Igor

    2015-11-01

    To compare the cleaning effectiveness and shaping ability of SafeSider, ProTaper Universal and Lightspeed rotary instruments during the preparation of curved root canals in extracted human teeth. A total of 63 roots with curved root canals were divided into three groups. Canals were prepared using SafeSider, ProTaper Universal or Lightspeed LSX. Using pre- and post-instrumentation radiographs, straightening of the canal curvatures and loss of working length were determined with a computer image analysis program. The amounts of debris at the apical 5 mm were quantified on the basis of a numerical evaluation scale. The data were analyzed statistically using the two-way analysis of variance (ANOVA). There was significantly more transportation among the Lightspeed LSX group compared to the SafeSider and ProTaper Universal groups only at the 4 mm level (p < 0.05). The ProTaper Universal instruments performed significantly faster than other groups. No significant differences were observed between the three engine-driven instruments with regards to debris removal. SafeSider, ProTaper Universal and Lightspeed LSX rotary instruments maintained the original canal curvature well at the apical 3 mm and were safe to use. No difference was found in cleaning efficacy and none rendered the apical part of the canal free of debris. SafeSider, ProTaper Universal and Lightspeed LSX rotary instruments are safe to use in curved root canals.

  13. Comparison of fracture resistance of simulated immature teeth with an open apex using Biodentine and composite resin: An in vitro study.

    PubMed

    Zhabuawala, Murtuza Saifuddin; Nadig, Roopa R; Pai, Veena S; Gowda, Yashwanth

    2016-01-01

    To evaluate the fracture resistance in simulated immature teeth that had been backfilled using composite resin and Biodentine after using Biodentine as an apical plug material immediately and after 3 months of aging. Sixty extracted human maxillary central incisors were simulated in an immature open apex. The roots of all the specimens were then standardized to a length of 10 mm and canals were instrumented to obtain the radicular dentin thickness around 1.5 mm. All the specimens were then randomly divided into three groups of twenty teeth each. Group I (control) - 4 mm apical plug of Biodentine backfilled with thermoplasticized gutta-percha. Group II - 4 mm apical plug of Biodentine and then backfilled with ParaCore. Group III - completely filled with Biodentine. Ten samples from each group were randomly divided into two subgroups. In subgroup A: Specimens were stored for 1 week. In subgroup B: Specimens were stored in phosphate-buffered saline solution for 3 months and were subjected to universal testing machine. Statistical analysis was done using one-way analysis. No significant difference in fracture resistance between the groups was observed when tested immediately. After 3 months of aging, only Biodentine group showed a significant reduction in fracture resistance without significant reduction with other two groups. Biodentine group has shown a drastic reduction in fracture resistance after 3 months of aging, and hence cannot be recommended as a reinforcement material in immature teeth with thin dentin walls.

  14. Effect of post-space treatment on retention of fiber posts in different root regions using two self-etching systems.

    PubMed

    Zhang, Ling; Huang, Li; Xiong, Yu; Fang, Ming; Chen, Ji-Hua; Ferrari, Marco

    2008-06-01

    The effect of post-space treatment on the retention of fiber posts in different root regions was evaluated using two self-etching systems. Post spaces were prepared in extracted premolars and then the root canals were subjected to one of the following post-space treatments: (i) water irrigation (control); (ii) etching with 35% phosphoric acid for 30 s; (iii) irrigation with 17% EDTA followed by 5.25% sodium hypochlorite (NaOCl); and (iv) ultrasonic agitation associated with 17% EDTA and 5.25% NaOCl irrigating solutions. The dentin surfaces were examined under scanning electron microscopy (SEM) after different post-space treatments. Fiber posts were then luted in the treated roots using resin cement with either Clearfil SE Bond or Clearfil DC Bond, and the thin-slice push-out test was performed. Scanning electron microscopy showed that all the post-space treatments tested were effective in removal of the smear layer of debris, or sealer/gutta-percha remnants, on the root canal. The apical push-out strength was affected by post-space treatment. Both 35% phosphoric acid etching and ultrasonic agitation in combination with EDTA/NaOCl irrigation improved the apical push-out strength of the fiber post, regardless of the type of self-etching system. A solo irrigation with an EDTA/NaOCl solution resulted in a lower apical push-out strength compared with the other two experimental groups.

  15. Chemomechanical preparation by hand instrumentation and by Mtwo engine-driven rotary files, an ex vivo study

    PubMed Central

    Krajczár, Károly; Tigyi, Zoltán; Papp, Viktória; Sára, Jeges; Tóth, Vilmos

    2012-01-01

    Objective: To compare the disinfecting efficacy of the sodium hypochlorite irrigation by root canal preparation with stainless steel hand files, taper 0.02 and nickel-titanium Mtwo files with taper 0.04-0.06. Study Design: 40 extracted human teeth were sterilized, and then inoculated with Enterococcus faecalis (ATCC 29212). After 6 day incubation time the root canals were prepared by hand with K-files (n=20) and by engine-driven Mtwo files (VDW, Munich, Germany) (n=20). Irrigation was carried out with 2.5% NaOCl in both cases. Samples were taken and determined in colony forming units (CFU) from the root canals before and after the preparation with instruments #25 and #35. Results: Significant reduction in bacterial count was determined after filing at both groups. The number of bacteria kept on decreasing with the extension of apical preparation diameter. There was no significant difference between the preparation sizes in the bacterial counts after hand or engine-driven instrumentation at the same apical size. Statistical analysis was carried out with Mann-Whitney test, paired t-test and independent sample t-test. Conclusions: Significant reduction in CFU was achieved after the root canal preparation completed with 2.5% NaOCl irrigation, both with stainless steel hand or nickel-titanium rotary files. The root canal remained slightly infected after chemo mechanical preparation in both groups. Key words:Chemomechanical preparation, root canal disinfection, nickel-titanium, conicity, greater taper, apical size. PMID:24558545

  16. Radiologically determined orthodontically induced external apical root resorption in incisors after non-surgical orthodontic treatment of class II division 1 malocclusion: a systematic review.

    PubMed

    Tieu, Long D; Saltaji, Humam; Normando, David; Flores-Mir, Carlos

    2014-07-23

    This study aims to critically evaluate orthodontically induced external apical root resorption (OIEARR) in incisors of patients undergoing non-surgical orthodontic treatment of class II division 1 malocclusion by a systematic review of the published data. An electronic search of two databases was performed; the bibliographies of relevant articles were also reviewed. Studies were included if they examined the amount of OIEARR in incisors produced during non-surgical orthodontic treatment of individuals with class II division I malocclusion in the permanent dentition. Individuals had no previous history of OIEARR, syndromes, pathologies, or general diseases. Study selections, risk of bias assessment, and data extraction were performed in duplicate. Eight studies of moderate methodological quality were finally included. An increased prevalence (65.6% to 98.1%) and mild to moderate severity of OIEARR (<4 mm and <1/3 original root) were reported. No sex difference in root resorption was found. For the maxillary incisors, there was no evidence that either the central or lateral incisor was more susceptible to OIEARR. A weak to moderate positive correlation between treatment duration and root resorption, and anteroposterior apical displacement and root resorption was found. Current limited evidence suggests that non-surgical comprehensive orthodontic treatment to correct class II division 1 malocclusions causes increased prevalence and severity of OIEARR the more the incisor roots are displaced and the longer this movement takes.

  17. A comparative evaluation of smear layer removal by using edta, etidronic acid, and maleic acid as root canal irrigants: An in vitro scanning electron microscopic study

    PubMed Central

    Kuruvilla, Aby; Jaganath, Bharath Makonahalli; Krishnegowda, Sahadev Chickmagaravalli; Ramachandra, Praveen Kumar Makonahalli; Johns, Dexton Antony; Abraham, Aby

    2015-01-01

    Aim: The purpose of this study is to evaluate and compare the efficacy of 17% EDTA, 18% etidronic acid, and 7% maleic acid in smear layer removal using scanning electron microscopic image analysis. Materials and Methods: Thirty, freshly extracted mandibular premolars were used. The teeth were decoronated to obtain working length of 17mm and instrumentation up to 40 size (K file) with 2.5% NaOCl irrigation between each file. The samples were divided into Groups I (17% ethylenediaminetetraacetic acid (EDTA)), II (18% etidronic acid), and III (7% maleic acid) containing 10 samples each. Longitudinal sectioning of the samples was done. Then the samples were observed under scanning electron microscope (SEM) at apical, middle, and coronal levels. The images were scored according to the criteria: 1. No smear layer, 2. moderate smear layer, and 3 heavy smear layer. Statistical Analysis: Data was analyzed statistically using Kruskal–Wallis analysis of variance (ANOVA) followed by Mann-Whitney U test for individual comparisons. The level for significance was set at 0.05. Results: The present study showed that all the three experimental irrigants removed the smear layer from different tooth levels (coronal, middle, and apical). Final irrigation with 7% maleic acid is more efficient than 17% EDTA and 18% etidronic acid in the removal of smear layer from the apical third of root canal. PMID:26069414

  18. Live Faecalibacterium prausnitzii Does Not Enhance Epithelial Barrier Integrity in an Apical Anaerobic Co-Culture Model of the Large Intestine.

    PubMed

    Maier, Eva; Anderson, Rachel C; Roy, Nicole C

    2017-12-12

    Appropriate intestinal barrier maturation during infancy largely depends on colonization with commensal bacteria. Faecalibacterium prausnitzii is an abundant obligate anaerobe that colonizes during weaning and is thought to maintain colonic health throughout life. We previously showed that F. prausnitzii induced Toll-like receptor 2 (TLR2) activation, which is linked to enhanced tight junction formation. Therefore, we hypothesized that F. prausnitzii enhances barrier integrity, an important factor in appropriate intestinal barrier maturation. In order to test metabolically active bacteria, we used a novel apical anaerobic co-culture system that allows the survival of both obligate anaerobic bacteria and oxygen-requiring intestinal epithelial cells (Caco-2). The first aim was to optimize the culture medium to enable growth and active metabolism of F. prausnitzii while maintaining the viability and barrier integrity, as measured by trans-epithelial electrical resistance (TEER), of the Caco-2 cells. This was achieved by supplementing the apical cell culture medium with bacterial culture medium. The second aim was to test the effect of F. prausnitzii on TEER across Caco-2 cell layers. Live F. prausnitzii did not improve TEER, which indicates that its benefits are not via altering tight junction integrity. The optimization of the novel dual-environment co-culturing system performed in this research will enable the investigation of new probiotics originating from indigenous beneficial bacteria.

  19. Live Faecalibacterium prausnitzii Does Not Enhance Epithelial Barrier Integrity in an Apical Anaerobic Co-Culture Model of the Large Intestine

    PubMed Central

    Maier, Eva; Anderson, Rachel C.; Roy, Nicole C.

    2017-01-01

    Appropriate intestinal barrier maturation during infancy largely depends on colonization with commensal bacteria. Faecalibacterium prausnitzii is an abundant obligate anaerobe that colonizes during weaning and is thought to maintain colonic health throughout life. We previously showed that F. prausnitzii induced Toll-like receptor 2 (TLR2) activation, which is linked to enhanced tight junction formation. Therefore, we hypothesized that F. prausnitzii enhances barrier integrity, an important factor in appropriate intestinal barrier maturation. In order to test metabolically active bacteria, we used a novel apical anaerobic co-culture system that allows the survival of both obligate anaerobic bacteria and oxygen-requiring intestinal epithelial cells (Caco-2). The first aim was to optimize the culture medium to enable growth and active metabolism of F. prausnitzii while maintaining the viability and barrier integrity, as measured by trans-epithelial electrical resistance (TEER), of the Caco-2 cells. This was achieved by supplementing the apical cell culture medium with bacterial culture medium. The second aim was to test the effect of F. prausnitzii on TEER across Caco-2 cell layers. Live F. prausnitzii did not improve TEER, which indicates that its benefits are not via altering tight junction integrity. The optimization of the novel dual-environment co-culturing system performed in this research will enable the investigation of new probiotics originating from indigenous beneficial bacteria. PMID:29231875

  20. Impaired left ventricular diastolic function is related to the formation of left ventricular apical thrombus in patients with acute anterior myocardial infarction.

    PubMed

    Choi, Ung Lim; Park, Jae-Hyeong; Sun, Byung Joo; Oh, Jin Kyung; Seong, Seok Woo; Lee, Jae-Hwan; Choi, Si Wan; Jeong, Jin-Ok; Kwon, In Sun; Seong, In-Whan

    2018-05-01

    Left ventricular (LV) apical thrombus is a clinically important complication which can cause systemic embolization in patients with anterior acute myocardial infarction (AMI). Systolic dysfunction has been a risk factor for developing LV apical thrombus in AMI patients. However, the role of diastolic dysfunction in the development of LV apical thrombus in these patients is still unknown. We performed this study to evaluate whether diastolic dysfunction can influence the development of LV apical thrombus in anterior AMI patients. We retrospectively analyzed all consecutive anterior AMI patients with available echocardiographic images within 1 month from January 2005 to April 2016. After gathering clinical characteristics from their medical records, systolic and diastolic functions were analyzed from digitally stored echocardiographic images. We included a total of 1045 patients (748 males, mean age 64 ± 12 years) with anterior AMI, and 494 (47%) were diagnosed as STEMI. The incidence of LV apical thrombus was 3.3% (34/1045). The LV apical thrombus group had larger LV diastolic dimension, larger LV diastolic and systolic volumes, and lower LVEF than the no LV thrombus group. The LV apical thrombus group showed higher mitral E velocity over mitral annular E' velocity ratio, an indicator of LV end-diastolic pressure (P < 0.001). In the LV apical thrombus group, the incidence of grade 2 diastolic dysfunction (32 vs 12%, P = 0.001) and grade 3 diastolic dysfunction (26 vs 2%, P < 0.001) were significantly higher than in the no LV apical thrombus group. The presence of more than grade 2 diastolic dysfunction, LVEF and presence of LV apical aneurysm were statistically significant factors associated with LV apical thrombus after the multivariate analysis. In conclusion, along with LV systolic dysfunction and LV apical aneurysm, LV diastolic dysfunction was also related with the presence of LV apical thrombus in patients with anterior AMI.

  1. Embryo as an active granular fluid: stress-coordinated cellular constriction chains

    NASA Astrophysics Data System (ADS)

    Holcomb, Michael; Gao, Guo-Jie; Thomas, Jeffrey; Blawzdziewicz, Jerzy

    2016-11-01

    Mechanical stress plays an intricate role in gene expression in individual cells and sculpting of developing tissues. Motivated by our observation of the cellular constriction chains (CCCs) during the initial phase of ventral furrow formation in the Drosophila melanogaster embryo, we propose an active granular fluid (AGF) model that provides valuable insights into cellular coordination in the apical constriction process. In our model, cells are treated as circular particles connected by a predefined force network, and they undergo a random constriction process in which the particle constriction probability P is a function of the stress exerted on the particle by its neighbors. We find that when P favors tensile stress, constricted particles tend to form chain-like structures. In contrast, constricted particles tend to form compact clusters when P favors compression. A remarkable similarity of constricted-particle chains and CCCs observed in vivo provides indirect evidence that tensile-stress feedback coordinates the apical constriction activity.

  2. Comparative analysis of efficacy and cleaning ability of hand and rotary devices for gutta-percha removal in root canal retreatment: an in vitro study.

    PubMed

    Reddy, Narender; Admala, Shilpa Reddy; Dinapadu, Sainath; Pasari, Srikanth; Reddy, Manoranjan P; Rao, M S Rama

    2013-07-01

    To evaluate the efficacy and cleaning ability of Hedstrom files, and ProTaper retreatment instruments in removing gutta-percha from root canals with and without xylene as solvent. Sixty extracted single rooted human teeth were selected and decoronated, straight access established working length determined 1 mm short of canal, chemomechanical preparation done and obturated with guttapercha and AH plus sealer. Samples were stored for 1 week in humidifier divided into four groups of 15 teeth each. • Group I: Hedstrom files without xylene. • Group II: Hedstrom files with xylene. • Group III: ProTaper retreatment instruments without xylene. • Group IV: ProTaper retreatment instruments with xylene. and the following criteria were assessed - Time taken for initial plunge of instrument into guttapercha. - Time taken for complete removal of gutta-percha to reach working length - Ability of H files and ProTaper retreatment files with/ without xylene to remove gutta-percha in coronal, middle and apical 1/3 of canal. The teeth were grooved in labiolingual cross section, observed under a steromicroscope and scored according to gutta-percha debris left in the canal. Results were evaluated using ANOVA test and multiple comparisons done using Scheffe test. The least time to reach working length was found with group IV followed by groups III, II and group I respectively. Also the fastest way to remove maximum gutta-percha was group IV followed by groups III, II, and I respectively with a statistically significant difference among all groups. Apical 1/3 has more amount of remaining gutta-percha debris than middle and coronal 1/3 in all groups. The amount of gutta-percha debris in apical 1/3 was least in group IV followed by groups III, II and I respectively. The better performance of ProTaper rotary instruments has been attributed to their special flute design which tends to pull gutta-percha coronally directing it toward orifice. Also the movements of engine driven instruments produce frictional heat which plasticises gutta-percha and aids in easy removal. Apical third of root canals showed more guttapercha debris compared to coronal and middle 1/3 and has been attributed to the greater anatomic variability and difficulty of instrumentation in the apical area. The existence of deep groves and depressions on dentine walls in this apical 1/3 make them less instrumented areas as it did be difficult to direct the file against the extreme root canal wall. The fastest technique to remove gutta-percha and the shortest time to reach working length was observed with ProTaper retreatment instruments with xylene followed by ProTaper retreatment files without xylene and Hedstrom files without xylene. After instrumentation for removal of gutta-percha, apical third was found to have more debris compared to coronal and middle 1/3 of the root canal.

  3. Cleaning effectiveness and shaping ability of rotary ProTaper compared with rotary GT and manual K-Flexofile.

    PubMed

    Liu, Sheng-Bo; Fan, Bin; Cheung, Gary S P; Peng, Bing; Fan, Ming-Wen; Gutmann, James L; Song, Ya-Ling; Fu, Qiang; Bian, Zhuan

    2006-12-01

    To compare the cleaning efficacy and shaping ability of engine-driven ProTaper and GT files, and manual preparation using K-Flexofile instruments in curved root canals of extracted human teeth. 45 canals of maxillary and mandibular molars with curvatures between 25 degrees and 40 degrees were divided into three groups. The groups were balanced with regard to the angle and the radius of canal curvature. Canals in each group were prepared to an apical size of 25 with either the rotary ProTaper or GT system, or manually with K-Flexofile using the modified double-flared technique. Irrigation was done with 2 mL 2.5% NaOCl after each instrument and, as the final rinse, 10 mL 2.5% NaOCl then 10 mL 17% EDTA and finally 5 mL distilled water. The double-exposure radiographic technique was used to examine for the presence of apical transportation. The time required to complete the preparation, as well as any change in working length after preparation were recorded. The roots were then grooved and split longitudinally. The amounts of debris and smear layer were evaluated at the apical, middle and coronal regions under the scanning electron microscope. Data were analyzed either parametrically with the F-test or non-parametrically using the Kruskal-Wallis test, where appropriate. Two GT files but none of the K-Flexofile and ProTaper instruments separated. For debris removal, the ProTaper group achieved a better result than GT (P < 0.05) but not the K-Flexofile group at all three regions (apical, middle and coronal). K-Flexofiles produced significantly less smear layer than ProTaper and GT files only in the middle third of the canal (P < 0.01). Both NiTi rotary instruments maintained the original canal shape better than the K-Flexofiles (P < 0.05) and required significantly less time to complete the preparation.

  4. Prevalence and morphometric analysis of three-rooted mandibular first molars in a Brazilian subpopulation

    PubMed Central

    Rodrigues, Clarissa Teles; de Oliveira-Santos, Christiano; Bernardineli, Norberti; Duarte, Marco Antonio Hungaro; Bramante, Clovis Monteiro; Minotti-Bonfante, Paloma Gagliardi; Ordinola-Zapata, Ronald

    2016-01-01

    ABSTRACT The knowledge of the internal anatomy of three-rooted mandibular molars may help clinicians to diagnose and plan the root canal treatment in order to provide adequate therapy when this variation is present. Objectives: To determine the prevalence of three-rooted mandibular molars in a Brazilian population using cone beam computed tomography (CBCT) and to analyze the anatomy of mandibular first molars with three roots through micro-CT. Material and Methods: CBCT images of 116 patients were reviewed to determine the prevalence of three-rooted first mandibular molars in a Brazilian subpopulation. Furthermore, with the use of micro-CT, 55 extracted three-rooted mandibular first molars were scanned and reconstructed to assess root length, distance between canal orifices, apical diameter, Vertucci's classification, presence of apical delta, number of foramina and furcations, lateral and accessory canals. The distance between the orifice on the pulp chamber floor and the beginning of the curvature and the angle of canal curvature were analyzed in the distolingual root. Data were compared using the Kruskal-Wallis test (α=0.05). Results: The prevalence of three-rooted mandibular first molars was of 2.58%. Mesial roots showed complex distribution of the root canal system in comparison to the distal roots. The median of major diameters of mesiobuccal, mesiolingual and single mesial canals were: 0.34, 0.41 and 0.60 mm, respectively. The higher values of major diameters were found in the distobuccal canals (0.56 mm) and the lower diameters in the distolingual canals (0.29 mm). The lowest orifice distance was found between the mesial canals (MB-ML) and the highest distance between the distal root canals (DB-DL). Almost all distal roots had one root canal and one apical foramen with few accessory canals. Conclusions: Distolingual root generally has short length, severe curvature and a single root canal with low apical diameter. PMID:27812625

  5. Establishing Apical Patency and its Effect on Endodontic Outcomes

    DTIC Science & Technology

    2012-06-01

    canal space and periodontal ligament. Establishing apical patency is controversial with only 50% of dental programs in the United States teaching the... periodontal ligament (PDL) (1) where a small file can passively continue through the apical foramen (2). Establishing apical patency is...teeth with apical periodontitis that will eventually heal demonstrate signs of healing at 1 year follow-up, and almost 50% are completely healed

  6. Apical closure of mature molar roots with the use of calcium hydroxide.

    PubMed

    Rotstein, I; Friedman, S; Katz, J

    1990-11-01

    Calcium hydroxide may induce apical root closure in affected mature teeth as well as in immature teeth. Once an apical hard tissue barrier is formed, a permanent root canal filling can be safely condensed. Two cases are described in which calcium hydroxide induced apical root closure in mature molar teeth where the apical constriction was lost because of chronic inflammatory process.

  7. Severe root resorption resulting from orthodontic treatment: Prevalence and risk factors

    PubMed Central

    Maués, Caroline Pelagio Raick; do Nascimento, Rizomar Ramos; Vilella, Oswaldo de Vasconcellos

    2015-01-01

    OBJECTIVE: To assess the prevalence of severe external root resorption and its potential risk factors resulting from orthodontic treatment. METHODS: A randomly selected sample was used. It comprised conventional periapical radiographs taken in the same radiology center for maxillary and mandibular incisors before and after active orthodontic treatment of 129 patients, males and females, treated by means of the Standard Edgewise technique. Two examiners measured and defined root resorption according to the index proposed by Levander et al. The degree of external apical root resorption was registered defining resorption in four degrees of severity. To assess intra and inter-rater reproducibility, kappa coefficient was used. Chi-square test was used to assess the relationship between the amount of root resorption and patient's sex, dental arch (maxillary or mandibular), treatment with or without extractions, treatment duration, root apex stage (open or closed), root shape, as well as overjet and overbite at treatment onset. RESULTS: Maxillary central incisors had the highest percentage of severe root resorption, followed by maxillary lateral incisors and mandibular lateral incisors. Out of 959 teeth, 28 (2.9%) presented severe root resorption. The following risk factors were observed: anterior maxillary teeth, overjet greater than or equal to 5 mm at treatment onset, treatment with extractions, prolonged therapy, and degree of apex formation at treatment onset. CONCLUSION: This study showed that care must be taken in orthodontic treatment involving extractions, great retraction of maxillary incisors, prolonged therapy, and/or completely formed apex at orthodontic treatment onset. PMID:25741825

  8. Severe root resorption resulting from orthodontic treatment: prevalence and risk factors.

    PubMed

    Maués, Caroline Pelagio Raick; do Nascimento, Rizomar Ramos; Vilella, Oswaldo de Vasconcellos

    2015-01-01

    To assess the prevalence of severe external root resorption and its potential risk factors resulting from orthodontic treatment. A randomly selected sample was used. It comprised conventional periapical radiographs taken in the same radiology center for maxillary and mandibular incisors before and after active orthodontic treatment of 129 patients, males and females, treated by means of the Standard Edgewise technique. Two examiners measured and defined root resorption according to the index proposed by Levander et al. The degree of external apical root resorption was registered defining resorption in four degrees of severity. To assess intra and inter-rater reproducibility, kappa coefficient was used. Chi-square test was used to assess the relationship between the amount of root resorption and patient's sex, dental arch (maxillary or mandibular), treatment with or without extractions, treatment duration, root apex stage (open or closed), root shape, as well as overjet and overbite at treatment onset. Maxillary central incisors had the highest percentage of severe root resorption, followed by maxillary lateral incisors and mandibular lateral incisors. Out of 959 teeth, 28 (2.9%) presented severe root resorption. The following risk factors were observed: anterior maxillary teeth, overjet greater than or equal to 5 mm at treatment onset, treatment with extractions, prolonged therapy, and degree of apex formation at treatment onset. This study showed that care must be taken in orthodontic treatment involving extractions, great retraction of maxillary incisors, prolonged therapy, and/or completely formed apex at orthodontic treatment onset.

  9. Nonsurgical root canal therapy of large cyst-like inflammatory periapical lesions and inflammatory apical cysts.

    PubMed

    Lin, Louis M; Ricucci, Domenico; Lin, Jarshen; Rosenberg, Paul A

    2009-05-01

    It is a general belief that large cyst-like periapical lesions and apical true cysts caused by root canal infection are less likely to heal after nonsurgical root canal therapy. Nevertheless, there is no direct evidence to support this assumption. A large cyst-like periapical lesion or an apical true cyst is formed within an area of apical periodontitis and cannot form by itself. Therefore, both large cyst-like periapical lesions and apical true cysts are of inflammatory and not of neoplastic origin. Apical periodontitis lesions, regardless of whether they are granulomas, abscesses, or cysts, fail to heal after nonsurgical root canal therapy for the same reason, intraradicular and/or extraradicular infection. If the microbial etiology of large cyst-like periapical lesions and inflammatory apical true cysts in the root canal is removed by nonsurgical root canal therapy, the lesions might regress by the mechanism of apoptosis in a manner similar to the resolution of inflammatory apical pocket cysts. To achieve satisfactory periapical wound healing, surgical removal of an apical true cyst must include elimination of root canal infection.

  10. Influenza A Virus Hemagglutinin and Neuraminidase Mutually Accelerate Their Apical Targeting through Clustering of Lipid Rafts

    PubMed Central

    Ohkura, Takashi; Momose, Fumitaka; Ichikawa, Reiko; Takeuchi, Kaoru

    2014-01-01

    ABSTRACT In polarized epithelial cells, influenza A virus hemagglutinin (HA) and neuraminidase (NA) are intrinsically associated with lipid rafts and target the apical plasma membrane for viral assembly and budding. Previous studies have indicated that the transmembrane domain (TMD) and cytoplasmic tail (CT) of HA and NA are required for association with lipid rafts, but the raft dependencies of their apical targeting are controversial. Here, we show that coexpression of HA with NA accelerated their apical targeting through accumulation in lipid rafts. HA was targeted to the apical plasma membrane even when expressed alone, but the kinetics was much slower than that of HA in infected cells. Coexpression experiments revealed that apical targeting of HA and NA was accelerated by their coexpression. The apical targeting of HA was also accelerated by coexpression with M1 but not M2. The mutations in the outer leaflet of the TMD and the deletion of the CT in HA and NA that reduced their association with lipid rafts abolished the acceleration of their apical transport, indicating that the lipid raft association is essential for efficient apical trafficking of HA and NA. An in situ proximity ligation assay (PLA) revealed that HA and NA were accumulated and clustered in the cytoplasmic compartments only when both were associated with lipid rafts. Analysis with mutant viruses containing nonraft HA/NA confirmed these findings. We further analyzed lipid raft markers by in situ PLA and suggest a possible mechanism of the accelerated apical transport of HA and NA via clustering of lipid rafts. IMPORTANCE Lipid rafts serve as sites for viral entry, particle assembly, and budding, leading to efficient viral replication. The influenza A virus utilizes lipid rafts for apical plasma membrane targeting and particle budding. The hemagglutinin (HA) and neuraminidase (NA) of influenza virus, key players for particle assembly, contain determinants for apical sorting and lipid raft association. However, it remains to be elucidated how lipid rafts contribute to the apical trafficking and budding. We investigated the relation of lipid raft association of HA and NA to the efficiency of apical trafficking. We show that coexpression of HA and NA induces their accumulation in lipid rafts and accelerates their apical targeting, and we suggest that the accelerated apical transport likely occurs by clustering of lipid rafts at the TGN. This finding provides the first evidence that two different raft-associated viral proteins induce lipid raft clustering, thereby accelerating apical trafficking of the viral proteins. PMID:24965459

  11. Treatment of Necrotic Teeth by Apical Revascularization: Meta-analysis.

    PubMed

    He, Ling; Zhong, Juan; Gong, Qimei; Kim, Sahng G; Zeichner, Samuel J; Xiang, Lusai; Ye, Ling; Zhou, Xuedong; Zheng, Jinxuan; Liu, Yongxing; Guan, Chenyu; Cheng, Bin; Ling, Junqi; Mao, Jeremy J

    2017-10-24

    Each year ~5.4 million children and adolescents in the United States suffer from dental infections, leading to pulp necrosis, arrested tooth-root development and tooth loss. Apical revascularization, adopted by the American Dental Association for its perceived ability to enable postoperative tooth-root growth, is being accepted worldwide. The objective of the present study is to perform a meta-analysis on apical revascularization. Literature search yielded 22 studies following PRISMA with pre-defined inclusion and exclusion criteria. Intraclass correlation coefficient was calculated to account for inter-examiner variation. Following apical revascularization with 6- to 66-month recalls, root apices remained open in 13.9% cases (types I), whereas apical calcification bridge formed in 47.2% (type II) and apical closure (type III) in 38.9% cases. Tooth-root lengths lacked significant postoperative gain among all subjects (p = 0.3472) or in subgroups. Root-dentin area showed significant increases in type III, but not in types I or II cases. Root apices narrowed significantly in types II and III, but not in type I patients. Thus, apical revascularization facilitates tooth-root development but lacks consistency in promoting root lengthening, widening or apical closure. Post-operative tooth-root development in immature permanent teeth represents a generalized challenge to regenerate diseased pediatric tissues that must grow to avoid organ defects.

  12. Live Faecalibacterium prausnitzii induces greater TLR2 and TLR2/6 activation than the dead bacterium in an apical anaerobic co-culture system.

    PubMed

    Maier, Eva; Anderson, Rachel C; Altermann, Eric; Roy, Nicole C

    2018-02-01

    Inappropriate activation of intestinal innate immune receptors, such as toll-like receptors (TLRs), by pathogenic bacteria is linked to chronic inflammation. In contrast, a "tonic" level of TLR activation by commensal bacteria is required for intestinal homeostasis. A technical challenge when studying this activation in vitro is the co-culturing of oxygen-requiring mammalian cells with obligate anaerobic commensal bacteria. To overcome this, we used a novel apical anaerobic co-culture system to successfully adapt a TLR activation assay to be conducted in conditions optimised for both cell types. Live Faecalibacterium prausnitzii, an abundant obligate anaerobe of the colonic microbiota, induced higher TLR2 and TLR2/6 activation than the dead bacterium. This enhanced TLR induction by live F. prausnitzii, which until now has not previously been described, may contribute to maintenance of gastrointestinal homeostasis. This highlights the importance of using physiologically relevant co-culture systems to decipher the mechanisms of action of live obligate anaerobes. © 2017 John Wiley & Sons Ltd.

  13. Periapical fluid RANKL and IL-8 are differentially regulated in pulpitis and apical periodontitis.

    PubMed

    Rechenberg, Dan-K; Bostanci, Nagihan; Zehnder, Matthias; Belibasakis, Georgios N

    2014-09-01

    The dental pulp space can become infected due to a breach in the surrounding hard tissues. This leads to inflammation of the pulp (pulpitis), soft tissue breakdown, and finally to bone loss around the root apex (apical periodontitis). The succession of the molecular events leading to apical periodontitis is currently not known. The main inflammatory mediator associated with neutrophil chemotaxis is interleukin-8 (IL-8), and with bone resorption the dyad of receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG). The levels of RANKL, OPG and IL-8 were studied in periapical tissue fluid of human teeth (n = 48) diagnosed with symptomatic irreversible pulpitis (SIP) and asymptomatic apical periodontitis (AAP). SIP represents the starting point, and AAP an established steady state of the disease. Periapical tissue fluid samples were collected using paper points and then evaluated using enzyme-linked immunosorbent assays (ELISAs). Target protein levels per case were calibrated against the corresponding total protein content, as determined fluorometrically. RANKL was expressed at significantly higher levels in SIP compared to AAP (P < 0.05), whereas OPG was under the detection limit in most samples. In contrast, IL-8 levels were significantly lower in SIP compared to AAP (P < 0.05). Spearman's correlation analysis between RANKL and IL-8 revealed a significantly (P < 0.05) negative correlation between the two measures (rho = -.44). The results of this study suggest that, in the development of apical periodontitis, periapical bone resorption signaling, as determined by RANKL, occurs prior to inflammatory cell recruitment signaling, as determined by IL-8. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Interleukin-6 and granulocyte-macrophage colony-stimulating factor in apical periodontitis: correlation with clinical and histologic findings of the involved teeth.

    PubMed

    Radics, T; Kiss, C; Tar, I; Márton, I J

    2003-02-01

    Apical periodontitis is characterized by the presence of immunocompetent cells producing a wide variety of inflammatory mediators. Releasing cytokines with long-range action, such as interleukin-6 (IL-6) and granulocyte-macrophage colony-stimulating factor (GM-CSF), apical periodontitis may induce changes in remote organs of the host. This study quantified the levels of IL-6 and GM-CSF in symptomatic and asymptomatic human periradicular lesions. Lesions were also characterized by size and histologic findings. Tissue samples were homogenized and supernatants were assayed using an enzyme-linked immunosorbent assay (ELISA). Correlations between cytokine levels and characteristic features (as single variables) of the lesions were analysed. There was a trend for higher levels of IL-6 and GM-CSF in symptomatic than in asymptomatic lesions, but the difference was not significant. Levels also tended to be higher in large than in small lesions, in polymorphonuclear (PMN) cell-rich than in PMN cell-poor samples, and in epithelialized than in non-epithelialized lesions. Significantly higher levels of IL-6 (778.1 +/- 220.5 pg/microg) and GM-CSF (363.3 +/- 98.4 pg/microg) were found in samples coincidentally possessing symptomatic and epithelialized features than in asymptomatic, small, PMN cell-poor, non-epithelialized lesions (IL-6: 45.2 +/- 13.1 pg/microg and GM-CSF: 135.1 +/- 26.4 pg/microg). These results suggest that symptomatic lesions containing epithelial cells represent an immunologically active stage of apical periodontitis, whereas asymptomatic, small, PMN cell-poor, non-epithelialized lesions represent healing apical lesions.

  15. Fitting the Elementary Rate Constants of the P-gp Transporter Network in the hMDR1-MDCK Confluent Cell Monolayer Using a Particle Swarm Algorithm

    PubMed Central

    Agnani, Deep; Acharya, Poulomi; Martinez, Esteban; Tran, Thuy Thanh; Abraham, Feby; Tobin, Frank; Ellens, Harma; Bentz, Joe

    2011-01-01

    P-glycoprotein, a human multidrug resistance transporter, has been extensively studied due to its importance to human health and disease. In order to understand transport kinetics via P-gp, confluent cell monolayers overexpressing P-gp are widely used. The purpose of this study is to obtain the mass action elementary rate constants for P-gp's transport and to functionally characterize members of P-gp's network, i.e., other transporters that transport P-gp substrates in hMDR1-MDCKII confluent cell monolayers and are essential to the net substrate flux. Transport of a range of concentrations of amprenavir, loperamide, quinidine and digoxin across the confluent monolayer of cells was measured in both directions, apical to basolateral and basolateral to apical. We developed a global optimization algorithm using the Particle Swarm method that can simultaneously fit all datasets to yield accurate and exhaustive fits of these elementary rate constants. The statistical sensitivity of the fitted values was determined by using 24 identical replicate fits, yielding simple averages and standard deviations for all of the kinetic parameters, including the efflux active P-gp surface density. Digoxin required additional basolateral and apical transporters, while loperamide required just a basolateral tranporter. The data were better fit by assuming bidirectional transporters, rather than active importers, suggesting that they are not MRP or active OATP transporters. The P-gp efflux rate constants for quinidine and digoxin were about 3-fold smaller than reported ATP hydrolysis rate constants from P-gp proteoliposomes. This suggests a roughly 3∶1 stoichiometry between ATP hydrolysis and P-gp transport for these two drugs. The fitted values of the elementary rate constants for these P-gp substrates support the hypotheses that the selective pressures on P-gp are to maintain a broad substrate range and to keep xenobiotics out of the cytosol, but not out of the apical membrane. PMID:22028772

  16. The barley MATE gene, HvAACT1, increases citrate efflux and Al(3+) tolerance when expressed in wheat and barley.

    PubMed

    Zhou, Gaofeng; Delhaize, Emmanuel; Zhou, Meixue; Ryan, Peter R

    2013-08-01

    Aluminium is toxic in acid soils because the soluble Al(3+) inhibits root growth. A mechanism of Al(3+) tolerance discovered in many plant species involves the release of organic anions from root apices. The Al(3+)-activated release of citrate from the root apices of Al(3+)-tolerant genotypes of barley is controlled by a MATE gene named HvAACT1 that encodes a citrate transport protein located on the plasma membrane. The aim of this study was to investigate whether expressing HvAACT1 with a constitutive promoter in barley and wheat can increase citrate efflux and Al(3+) tolerance of these important cereal species. HvAACT1 was over-expressed in wheat (Triticum aestivum) and barley (Hordeum vulgare) using the maize ubiquitin promoter. Root apices of transgenic and control lines were analysed for HvAACT1 expression and organic acid efflux. The Al(3+) tolerance of transgenic and control lines was assessed in both hydroponic solution and acid soil. Increased HvAACT1 expression in both cereal species was associated with increased citrate efflux from root apices and enhanced Al(3+) tolerance, thus demonstrating that biotechnology can complement traditional breeding practices to increase the Al(3+) tolerance of important crop plants.

  17. Commensal-Associated Molecular Patterns Induce Selective Toll-Like Receptor-Trafficking from Apical Membrane to Cytoplasmic Compartments in Polarized Intestinal Epithelium

    PubMed Central

    Cario, Elke; Brown, Dennis; McKee, Mary; Lynch-Devaney, Kathryn; Gerken, Guido; Podolsky, Daniel K.

    2002-01-01

    Commensal-associated molecular patterns, the major products of nonpathogenic bacteria, are present at high concentrations at the apical surface of the intestinal epithelium. However, the nature of the interaction of commensal-associated molecular patterns with the lumenal surface of the epithelium has not been defined. We have recently demonstrated that intestinal epithelial cells constitutively express several Toll-like receptors (TLRs) in vitro and in vivo that seem to be the key receptors responsible for immune cell activation in response to various bacterial products. In this study we characterize the subcellular distribution of two major TLRs, TLR2 and TLR4, and their ligand-specific dynamic regulation in the model human intestinal epithelial cell line T84. Immunocytochemical studies indicate that TLR2 and TLR4 are constitutively expressed at the apical pole of differentiated T84 cells. After stimulation with lipopolysaccharide or peptidoglycan, TLRs selectively traffic to cytoplasmic compartments near the basolateral membrane. Thus, we demonstrate that TLRs are positioned at the apical pole where they are poised to monitor the sensitive balance of the lumenal microbial array. The results of this dynamic epithelial surveillance can then be conveyed to the underlying cell populations of the lamina propria via these innate immune pattern recognition receptors. PMID:11786410

  18. Actin Turnover-Mediated Gravity Response in Maize Root Apices

    PubMed Central

    Mancuso, Stefano; Barlow, Peter W; Volkmann, Dieter

    2006-01-01

    The dynamic actin cytoskeleton has been proposed to be linked to gravity sensing in plants but the mechanistic understanding of these processes remains unknown. We have performed detailed pharmacological analyses of the role of the dynamic actin cytoskeleton in gravibending of maize (Zea mays) root apices. Depolymerization of actin filaments with two drugs having different mode of their actions, cytochalasin D and latrunculin B, stimulated root gravibending. By contrast, drug-induced stimulation of actin polymerization and inhibition of actin turnover, using two different agents phalloidin and jasplakinolide, compromised the root gravibending. Importantly, all these actin drugs inhibited root growth to similar extents suggesting that high actin turnover is essential for the gravity-related growth responses rather than for the general growth process. Both latrunculin B and cytochalasin D treatments inhibited root growth but restored gravibending of the decapped root apices, indicating that there is a strong potential for effective actin-mediated gravity sensing outside the cap. This elusive gravity sensing outside the root cap is dependent not only on the high rate of actin turnover but also on weakening of myosin activities, as general inhibition of myosin ATPases induced stimulation of gravibending of the decapped root apices. Collectively, these data provide evidence for the actin turnover-mediated gravity sensing outside the root cap. PMID:19521476

  19. Epithelial Folding Driven by Apical or Basal-Lateral Modulation: Geometric Features, Mechanical Inference, and Boundary Effects.

    PubMed

    Wen, Fu-Lai; Wang, Yu-Chiun; Shibata, Tatsuo

    2017-06-20

    During embryonic development, epithelial sheets fold into complex structures required for tissue and organ functions. Although substantial efforts have been devoted to identifying molecular mechanisms underlying epithelial folding, far less is understood about how forces deform individual cells to sculpt the overall sheet morphology. Here we describe a simple and general theoretical model for the autonomous folding of monolayered epithelial sheets. We show that active modulation of intracellular mechanics along the basal-lateral as well as the apical surfaces is capable of inducing fold formation in the absence of buckling instability. Apical modulation sculpts epithelia into shallow and V-shaped folds, whereas basal-lateral modulation generates deep and U-shaped folds. These characteristic tissue shapes remain unchanged when subject to mechanical perturbations from the surroundings, illustrating that the autonomous folding is robust against environmental variabilities. At the cellular scale, how cells change shape depends on their initial aspect ratios and the modulation mechanisms. Such cell deformation characteristics are verified via experimental measurements for a canonical folding process driven by apical modulation, indicating that our theory could be used to infer the underlying folding mechanisms based on experimental data. The mechanical principles revealed in our model could potentially guide future studies on epithelial folding in diverse systems. Copyright © 2017. Published by Elsevier Inc.

  20. Survival of the Apical Papilla and Its Resident Stem Cells in a Case of Advanced Pulpal Necrosis and Apical Periodontitis.

    PubMed

    Chrepa, Vanessa; Pitcher, Brandon; Henry, Michael A; Diogenes, Anibal

    2017-04-01

    Apical papilla represents a source of an enriched mesenchymal stem cell (MSC) population (stem cells of the apical papilla [SCAPs]) that modulates root development and may participate in regenerative endodontic procedures in immature teeth with pulp necrosis. The characteristics and phenotype of this tissue in the presence of inflammation are largely unknown. The purpose of this study was to characterize a human apical papilla sample that was isolated from an immature tooth with pulp necrosis and apical periodontitis. Inflamed periapical tissue that included part of the apical papilla (apical papilla clinical sample [CS]) was collected from an immature mandibular premolar previously diagnosed with pulp necrosis and apical periodontitis during an apexification procedure. Harvested cells from this tissue (SCAP CS) were compared with inflamed periapical progenitor cells (IPAPCs) and normal SCAP (SCAP-RP89) in flow cytometry and quantitative osteogenesis experiments. Part of the issue was further processed for immunohistochemistry and compared with apical papilla and coronal pulp sections from normal immature teeth as well as inflamed periapical tissues from mature teeth. Similar to SCAP-RP89, 96.6% of the SCAP CS coexpressed the MSC markers CD73, CD90, and CD105, whereas only 66.3% of IPAPCs coexpressed all markers. The SCAP CS showed a significantly greater mineralization potential than both SCAP-RP89 and IPAPCs. Finally, immunohistochemical analysis revealed moderate infiltration of cells expressing the inflammatory markers CD45/68 in the apical papilla CS and prominent CD24, CD105, and von Willebrand factor expression. Under inflammatory conditions, human apical papilla was found moderately inflamed with retained SCAP vitality and stemness and increased osteogenic and angiogenesis potential. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  1. SAX-7/L1CAM and HMR-1/cadherin function redundantly in blastomere compaction and non-muscle myosin accumulation during C. elegans gastrulation

    PubMed Central

    Grana, Theresa M.; Cox, Elisabeth A.; Lynch, Allison M.; Hardin, Jeff

    2010-01-01

    Gastrulation is the first major morphogenetic movement in development, and requires dynamic regulation of cell adhesion and the cytoskeleton. C. elegans gastrulation begins with the migration of the two endodermal precursors, Ea and Ep, from the surface of the embryo into the interior. Ea/Ep migration provides a relatively simple system to examine the intersection of cell adhesion, cell signaling, and cell movement. Ea/Ep ingression depends on correct cell fate specification and polarization, apical myosin accumulation, and Wnt activated actomyosin contraction that drives apical constriction and ingression (Lee et al., 2006; Nance et al., 2005). Here, we show that Ea/Ep ingression also requires the function of either HMR-1/cadherin or SAX-7/L1CAM. Both cadherin complex components and L1CAM are localized at all sites of cell-cell contact during gastrulation. Either system is sufficient for Ea/Ep ingression, but loss of both together leads to a failure of apical constriction and ingression. Similar results are seen with isolated blastomeres. Ea/Ep are properly specified and appear to display correct apical-basal polarity in sax-7(eq1); hmr-1(RNAi) embryos. Significantly, in sax-7(eq1); hmr-1(RNAi) embryos Ea and Ep fail to accumulate myosin (NMY-2::GFP) at their apical surfaces, but in either sax-7(eq1) or hmr-1(RNAi) embryos, apical myosin accumulation is comparable to wildtype. Thus, the cadherin and L1CAM adhesion systems are redundantly required for localized myosin accumulation, and hence for actomyosin contractility during gastrulation. We also show that sax-7 and hmr-1 function are redundantly required for Wnt-dependent spindle polarization during division of the ABar blastomere, indicating that these cell surface proteins redundantly regulate multiple developmental events in early embryos. PMID:20515680

  2. Study of some invasiveness markers as pathogenic factors in oral pseudoepitheliomatous hyperplasia.

    PubMed

    Pascu, Roxana Maria; Mărgăritescu, Claudiu; CrăiŢoiu, Monica Mihaela; Florescu, Alma Maria; Croitoru, Ileana Cristiana; Bobic, Adelina Gabriela; Pătru, Ciprian LaurenŢiu; Mălăescu, Gheorghe Dan; CrăiŢoiu, Ştefania

    2016-01-01

    Pseudoepitheliomatous hyperplasia is a benign reactivated epithelial lesion secondary to another pathology, whose incidence is difficult to establish. There still exist controversies regarding the origin and pathogenesis of these lesions. For this purpose, we performed an immuno-histochemical study upon 20 cases of oral pseudoepitheliomatous hyperplasia associated with inflammatory and neoplastic conditions, investigating a series of markers with a possible pathogenic potential in developing this type of lesions. Thus, the immunoreactivity study for β-catenin showed the presence of a membrane reactivity in all the stratum spinosum and a predominantly cytoplasmatic reactivity, more rarely a nuclear one, in the cells of the basal stratum cells, especially in the epithelial apices that descend deeply in the chorion. Instead, in the case of vimentin, the reactivity was present only in the epithelial apices, especially in the peripheral cells, in comparison to the central ones, and especially in the cases where the epithelial apices descended deeply in the sublesional chorion. Moreover, we observed that the MMP9 reactivity in pseudoepitheliomatous hyperplasia lesions was present in the cells at the epithelium-chorion interface and especially in the epithelial apices that descend deeply into the chorion, and also in the epithelial chorion and networks. The study for CXCR4 immuno-reactivity showed a good reactivity in almost all layers of this hyperplastic lesion, with a maximum reactivity especially inside the epithelial apices that descend deeply in the sublesional chorion. Such an immunoprofile suggests the ability of the oral epithelial cells to undergo an epithelial mesenchymal transition process, thus acquiring mesenchymal characteristics through which it deeply migrates in the subadjacent chorion and contributes to the formation of epithelial apices in pseudoepitheliomatous hyperplasia. Moreover, the invasive ability of these lesions is also given by the average quantity of matrix metalloproteinases present in the epithelium-chorion interface determined by the activation of CXCR4 receptors at this level.

  3. Levels of oxidative stress biomarkers and bone resorption regulators in apical periodontitis lesions infected by Epstein-Barr virus.

    PubMed

    Jakovljevic, A; Andric, M; Nikolic, N; Coric, V; Krezovic, S; Carkic, J; Knezevic, A; Beljic-Ivanovic, K; Pljesa-Ercegovac, M; Miletic, M; Soldatovic, I; Radosavljevic, T; Jovanovic, T; Simic, T; Ivanovic, V; Milasin, J

    2018-06-01

    To investigate whether apical periodontitis lesions infected by Epstein-Barr virus (EBV) exhibit higher levels of oxidative stress biomarkers [8-hydroxydeoxyguanosine (8-OHdG) and oxidized glutathione (GSSG)] and bone resorption regulators [receptor activator of nuclear factor (NF-κB) ligand (RANKL) and osteoprotegerin (OPG)] compared to EBV-negative periapical lesions and healthy pulp tissues. The experimental group consisted of 30 EBV-positive and 30 EBV-negative periapical lesions collected in conjunction with apicoectomy. The pulp tissues of 20 impacted third molars were used as healthy controls. The qualitative and quantitative analysis of EBV was performed by nested and real-time polymerase chain reaction (PCR), respectively. The levels of RANKL and OPG were analysed by reverse transcriptase real-time PCR. The levels of 8-OHdG and GSSG were determined by enzyme-linked immunosorbent assay (ELISA). Mann-Whitney U-test and Spearman's correlation were used for statistical analysis. The levels of RANKL, OPG, 8-OHdG and GSSG were significantly higher in apical periodontitis lesions compared to healthy pulp controls (P = 0.001, P < 0.001, P < 0.001 and P < 0.05, respectively). RANKL and OPG mRNA expression was significantly higher in EBV-positive compared to EBV-negative periapical lesions (P < 0.05). There was no significant correlation between EBV copy numbers and levels of RANKL, OPG, 8OH-dG and GSSG in apical periodontitis. Levels of bone resorption regulators and oxidative stress biomarkers were increased in apical periodontitis compared to healthy pulp tissues. EBV-positive periapical lesions exhibited higher levels of RANKL and OPG compared to EBV-negative periapical lesions. EBV may contribute to progression of apical periodontitis via enhanced production of bone resorption regulators. © 2018 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  4. Apical closure device for full-percutaneous transapical valve implantation: stress-test in an animal model†.

    PubMed

    Ferrari, Enrico; Demertzis, Stefanos; Angelella, Jennifer; Berdajs, Denis; Tozzi, Piergiorgio; Moccetti, Tiziano; Maisano, Francesco; von Segesser, Ludwig K

    2017-05-01

    Transapical valve implantation is traditionally performed through a left antero-lateral mini-thoracotomy. A self-expandable apical closure device has recently been developed for full-percutaneous transapical valve implantation. We performed haemodynamics stress-tests on an animal model to evaluate the sealing properties. Under general anaesthesia 5 pigs (mean weight: 67 ± 6 Kg) received full heparinization (100 IU/Kg; activated clotting time >250 s and, through inferior mini-sternotomies, 21-Fr introducer sheaths for transapical aortic valve implantation (outer diameter: 25-Fr) were placed over-the-wire in the apexes. Delivery-catheters carrying folded occluders (SAFEX TM final design) were inserted in the introducer sheaths and plugs were then deployed under fluoroscopic guidance. Phase 1: after protamine injection, apical bleeding was monitored for 1 h with standard haemodynamics condition. Phase 2: we induced systemic hypertension with adrenaline infusion to test the sealing properties under stress. Animals were sacrificed after Phase 2 and hearts were removed and inspected. Five plugs were successfully introduced and deployed in 5 pig hearts. Plugs provided good apical sealing in each animal and a mean of 7 ± 4 ml of blood lost per animal was collected during Phase 1: haemodynamics remained stable and no plug dislodgement was detected (mean blood pressure: 52 ± 9 mmHg). During Phase 2, mean systolic and diastolic peak levels reached 268 ± 24 mmHg and 175 ± 17 mmHg, respectively, without plug dislodgment or bleeding. Post-mortem inspection showed good plug deployment and fixation without myocardial damage. The new apical occluder seals large-sized apical access sites in animal models also during induced systemic hypertension. This pilot study is a further step towards full-percutaneous transapical valve procedures in the clinical setting. © The Author 2017. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.

  5. Microscopic Examination of Oral Sinus Tracts and Their Associated Periapical Lesions,

    DTIC Science & Technology

    1983-05-01

    granulomas (chronic apical periodontitis ), 4 periapical cysts, and 7 periapical abscesses (Table 2). Only 2 of the 15 biopsied periapical lesions had a...granuloma) (2) or an acute apical abscess (2). Ingle (4) believes that the development of a "suppurative apical periodontitis " from an acute alveolar...radio- lucencies are consistent with a chronic apical periodontitis . With an acute apical abscess , all that is usually radiographically visible is a

  6. Association of human herpesvirus 6 subtypes with symptomatic apical periodontitis.

    PubMed

    Hernádi, Katinka; Csoma, Eszter; Adám, Balázs; Szalmás, Anita; Gyöngyösi, Eszter; Veress, György; Ildikó-Márton; Kónya, József

    2011-09-01

    The occurrence of human herpesvirus (HHV) 6 subtypes A and B in apical periodontitis was determined. The relationship of HHV-6 subtypes to other disease associated herpesviruses, i.e., Epstein-Barr virus (EBV) and human cytomegalovirus, was also investigated. Forty apical periodontitis samples (17 symptomatic and 23 asymptomatic) and 40 healthy pulp control samples were collected. Nested polymerase chain reaction was used to detect HHV-6 DNA. HHV-6 DNA was observed in significantly higher frequencies in apical periodontitis samples than in control samples (20% vs. 2.5%; P = .03). Further classification of apical lesions revealed that subtype B of HHV-6 was significantly associated with large-sized and symptomatic lesions (P < .01). Thirty-one apical lesions (77%) harbored ≥1 of the tested herpesviruses: EBV was the most frequent herpesvirus (72.5%) in apical periodontitis, followed by HHV-6 (20%). Our findings suggest that EBV and HHV-6B infections can be associated with symptomatic apical periodontitis. Copyright © 2011 Mosby, Inc. All rights reserved.

  7. Loss of Gα12/13 exacerbates apical area dependence of actomyosin contractility

    PubMed Central

    Xie, Shicong; Mason, Frank M.; Martin, Adam C.

    2016-01-01

    During development, coordinated cell shape changes alter tissue shape. In the Drosophila ventral furrow and other epithelia, apical constriction of hundreds of epithelial cells folds the tissue. Genes in the Gα12/13 pathway coordinate collective apical constriction, but the mechanism of coordination is poorly understood. Coupling live-cell imaging with a computational approach to identify contractile events, we discovered that differences in constriction behavior are biased by initial cell shape. Disrupting Gα12/13 exacerbates this relationship. Larger apical area is associated with delayed initiation of contractile pulses, lower apical E-cadherin and F-actin levels, and aberrantly mobile Rho-kinase structures. Our results suggest that loss of Gα12/13 disrupts apical actin cortex organization and pulse initiation in a size-dependent manner. We propose that Gα12/13 robustly organizes the apical cortex despite variation in apical area to ensure the timely initiation of contractile pulses in a tissue with heterogeneity in starting cell shape. PMID:27489340

  8. Signaling-Dependent Control of Apical Membrane Size and Self-Renewal in Rosette-Stage Human Neuroepithelial Stem Cells.

    PubMed

    Medelnik, Jan-Philip; Roensch, Kathleen; Okawa, Satoshi; Del Sol, Antonio; Chara, Osvaldo; Mchedlishvili, Levan; Tanaka, Elly M

    2018-06-05

    In the developing nervous system, neural stem cells are polarized and maintain an apical domain facing a central lumen. The presence of apical membrane is thought to have a profound influence on maintaining the stem cell state. With the onset of neurogenesis, cells lose their polarization, and the concomitant loss of the apical domain coincides with a loss of the stem cell identity. Little is known about the molecular signals controlling apical membrane size. Here, we use two neuroepithelial cell systems, one derived from regenerating axolotl spinal cord and the other from human embryonic stem cells, to identify a molecular signaling pathway initiated by lysophosphatidic acid that controls apical membrane size and consequently controls and maintains epithelial organization and lumen size in neuroepithelial rosettes. This apical domain size increase occurs independently of effects on proliferation and involves a serum response factor-dependent transcriptional induction of junctional and apical membrane components. Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.

  9. Tolerance limit value of brightness and contrast adjustment on digitized radiographs

    NASA Astrophysics Data System (ADS)

    Utami, S. N.; Kiswanjaya, B.; Syahraini, S. I.; Ustriyana, P.

    2017-08-01

    The aim of this study was to measure the tolerance limit value of brightness and contrast adjustment on digitized radiograph with apical periodontitis and early apical abscess. Brightness and contrast adjustment on 60 periapical radiograph with apical periodontitis and early apical abscess made by 2 observers. Reliabilities tested by Cohen’s Kappa Coefficient and significance tested by wilcoxon test. Tolerance limit value of brightness and contrast adjustment for apical periodontitis is -5 and +5, early apical abscess is -10 and +10, and both is -5 and +5. Brightness and contrast adjustment which not appropriate can alter the evaluation and differential diagnosis of periapical lesion.

  10. Clathrin and AP1 are required for apical sorting of glycosyl phosphatidyl inositol-anchored proteins in biosynthetic and recycling routes in Madin-Darby canine kidney cells.

    PubMed

    Castillon, Guillaume A; Burriat-Couleru, Patricia; Abegg, Daniel; Criado Santos, Nina; Watanabe, Reika

    2018-03-01

    Recently, studies in animal models demonstrate potential roles for clathrin and AP1 in apical protein sorting in epithelial tissue. However, the precise functions of these proteins in apical protein transport remain unclear. Here, we reveal mistargeting of endogenous glycosyl phosphatidyl inositol-anchored proteins (GPI-APs) and soluble secretory proteins in Madin-Darby canine kidney (MDCK) cells upon clathrin heavy chain or AP1 subunit knockdown (KD). Using a novel directional endocytosis and recycling assay, we found that these KD cells are not only affected for apical sorting of GPI-APs in biosynthetic pathway but also for their apical recycling and basal-to-apical transcytosis routes. The apical distribution of the t-SNARE syntaxin 3, which is known to be responsible for selective targeting of various apical-destined cargo proteins in both biosynthetic and endocytic routes, is compromised suggesting a molecular explanation for the phenotype in KD cells. Our results demonstrate the importance of biosynthetic and endocytic routes for establishment and maintenance of apical localization of GPI-APs in polarized MDCK cells. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. Effectiveness of Rotary Endodontic Instruments on Smear Layer Removal in Root Canals of Primary Teeth: A Scanning Electron Microscopy Study.

    PubMed

    Subramaniam, Priya; Girish Babu, K L; Tabrez, T A

    2016-01-01

    The present SEM study was undertaken to evaluate the effect of root canal instrumentation using both manual and rotary files in the root canals of primary anterior teeth. Thirty freshly extracted primary maxillary incisors were divided into 3 groups of 10 teeth each. In Group I, root canals were instrumented with rotary NiTi files; in Group II, the root canals were instrumented using manual NiTi K files and; in Group III, manual instrumentation was done with stainless steel K files. Longitudinal sections were prepared and processed for observation under SEM at the coronal, middle and apical thirds. Scoring of smear layer was done according to Hulsmann and the data obtained was subjected to statistical analysis. Rotary files cleaned the coronal and middle thirds of root canals more effectively. Statistically there was no significant difference between the groups. Lowest score of 2.6 in the apical third of root canals was seen with hand NiTi files. Rotary instrumentation was as effective as manual instrumentation in removal of smear layer in the root canals of primary anterior teeth.

  12. Apical extrusion of Enterococcus faecalis using three different rotary instrumentation techniques: an in vitro study.

    PubMed

    Taneja, Sonali; Kumari, Manju; Barua, Madhumita; Dudeja, Chetna; Malik, Meeta

    2015-01-01

    To compare the apical extrusion of Enterococcus faecalis after instrumentation with three different Ni-Ti rotary instruments- An in vitro study. In vitro study Methods and Material: Forty freshly extracted mandibular premolars were mounted in bacteria collection apparatus and root canals were contaminated with a suspension of Enterococcus faecalis. The contaminated teeth were divided into 4 groups of 10 teeth each according to rotary system used for instrumentation: Group1: Hyflex files, Group 2: GTX files, Group 3: Protaper files and Group 4: control group (no instrumentation). Bacteria extruded after preparations were collected into vials and microbiological samples were incubated in BHI broth for 24 hrs. The colony forming units were determined for each sample. Statistical analysis was done using one way ANOVA followed by post hoc independent " t" test. GTX files extruded least amount of bacteria followed by Hyflex files. Maximum extrusion of E. faecalis was seen in rotary Protaper group. Least amount of extrusion was seen with GTX files followed by Hyflex files and then rotary Protaper system.

  13. [Bonding interfaces of three kinds of cements and root canal dentin: a scanning electron microscope observation].

    PubMed

    Chen, Lei; Lei, Hui-yun; Xu, Guo-fu; Liang, Xiao-peng; Li, Ji-jia

    2010-04-01

    To compare the bonding properties of three kinds of cements by observing the bonding inteffaces of cements and root canal dentin. 15 extracted mandibular premolars were divided into 3 groups, and were cemented by Rely X luting, Panavia F and Paracore 5 mL, respectively. Each tooth was sectioned into two parts and the dentin-cement interfaces at the coronal, middle and apical parts of the fiber post were oberved by scanning electron microscope (SEM). The length of hybrid layer was also recorded. Hybrid layer was not clearly found in group one, which could be seen on the dentin-cement interfaces of group two and three. Resin tags and lateral adhesives were also observed in group three. From the apical to the coronal part, microgaps seemed gradually smaller in group one, while the hybrid layer became thicker in both group two and three. The total-etch resin cement bounds tightly with dentin, and owns a more superior bonding property than self-etch resin cement and resin modified glass ionomer cement.

  14. In vitro study of the variable square pulse Er:YAG laser cutting efficacy for apicectomy.

    PubMed

    Grgurević, Josko; Grgurević, Lovro; Miletić, Ivana; Karlović, Zoran; Krmek, Silvana Jukić; Anić, Ivica

    2005-06-01

    Variable square pulse (VSP) Er:YAG laser should be quicker than older Er:YAG lasers. The objectives were: (1) comparison of VSP laser and mechanical handpiece efficacy for apicectomy and (2) determination of optimal pulse width/energy/frequency combination. Sixty extracted, single-rooted mature human teeth with round apical parts were instrumented, root filled, cleaned, and divided into four groups. Apical 2 mm of each root were apicectomized with mechanical handpiece and Er:YAG laser with three different settings (LaserA = 200 mJ/300 microseconds/ 8 Hz; LaserB = 200 mJ/100 microseconds/8 Hz; LaserC = 380 mJ/100 microseconds/20 Hz). Timing results were statistically compared. LaserC was the most efficient setting. Differences between groups were significant except between LaserC-Mechanical and LaserA-LaserC (P < 0.05). VSP Er:YAG laser used for apicectomy is slower by a factor of 7-31 than mechanical handpiece, but treatment outcome is acceptable. Optimal settings for apicectomy with VSP laser are 380 mJ/100 microseconds/20 Hz. Copyright 2005 Wiley-Liss, Inc.

  15. Cleaning Effectiveness of a Reciprocating Single-file and a Conventional Rotary Instrumentation System

    PubMed Central

    de Carvalho, Fredson Marcio Acris; Gonçalves, Leonardo Cantanhede de Oliveira; Marques, André Augusto Franco; Alves, Vanessa; Bueno, Carlos Eduardo da Silveira; De Martin, Alexandre Sigrist

    2016-01-01

    Objective: To compare cleaning effectiveness by histological analysis of a reciprocating single-file system with ProTaper rotary instruments during the preparation of curved root canals in extracted teeth. Methods: A total of 40 root canals with curvatures ranging between 20 - 40 degrees were divided into two groups of 20 canals. Canals were prepared to the following apical sizes: Reciproc size 25 (n=20); ProTaper: F2 (n=20). The normal distribution of data was tested by the Kolmogorov-Smirnov test and the values obtained for the test (Mann-Whitney U test, P < .05) were statistically analyzed using the GraphPad InStat for the Mac OS software (GraphPad Software, La Jolla, CA, USA). Results: There were no significant differences in remaining debris (P > .05) between the two groups. Conclusion: The application of reciprocating motion during instrumentation did not result in increased debris when compared with continuous rotation motion, even in the apical part of curved canals. Both instruments resulted in debris in the canal lumen, irrespective of the movement kinematics applied. PMID:28217185

  16. Apical root resorption in patients treated with comprehensive orthodontics.

    PubMed

    Harris, E F; Boggan, B W; Wheeler, D A

    2001-01-01

    External apical root resorption (EARR) is a common--but seldom extreme--consequence of orthodontic treatment. Incisors are most at risk, perhaps because of their single roots and because they typically are moved farther than other teeth. We followed a cohort of patients (n = 153) treated with comprehensive orthodontics. EARR was scored on the upper incisors with a qualitative five-grade ordinal scale. There was no EARR at the start of treatment, but most (> 80%) exhibited slight-to-moderate EARR by the end of treatment (i.e., a loss of 1-2 mm). Cases treated with premolar extractions experienced more EARR because their incisors were retracted farther; however, the sum of the effects of patients' sex and age, and severity of the malocclusion, and the kind of mechanics used accounts for little of the overall variation in EARR. Instead, it appears that genetically-based inter-individual variation in susceptibility to EARR is the most influential factor. Research should be directed at understanding the biochemical nature of susceptibility so prospective patients can be screened to identify those at particular risk.

  17. Forskolin-induced apical membrane insertion of virally expressed, epitope-tagged CFTR in polarized MDCK cells.

    PubMed

    Howard, M; Jiang, X; Stolz, D B; Hill, W G; Johnson, J A; Watkins, S C; Frizzell, R A; Bruton, C M; Robbins, P D; Weisz, O A

    2000-08-01

    Channel gating of the cystic fibrosis transmembrane conductance regulator (CFTR) is activated in response to cAMP stimulation. In addition, CFTR activation may also involve rapid insertion of a subapical pool of CFTR into the plasma membrane (PM). However, this issue has been controversial, in part because of the difficulty in distinguishing cell surface vs. intracellular CFTR. Recently, a fully functional, epitope-tagged form of CFTR (M2-901/CFTR) that can be detected immunologically in nonpermeabilized cells was characterized (Howard M, Duvall MD, Devor DC, Dong J-Y, Henze K, and Frizzell RA. Am J Physiol Cell Physiol 269: C1565-C1576, 1995; and Schultz BD, Takahashi A, Liu C, Frizzell RA, and Howard M. Am J Physiol Cell Physiol 273: C2080-C2089, 1997). We have developed replication-defective recombinant adenoviruses that express M2-901/CFTR and used them to probe cell surface CFTR in forskolin (FSK)-stimulated polarized Madin-Darby canine kidney (MDCK) cells. Virally expressed M2-901/CFTR was functional and was readily detected on the apical surface of FSK-stimulated polarized MDCK cells. Interestingly, at low multiplicity of infection, we observed FSK-stimulated insertion of M2901/CFTR into the apical PM, whereas at higher M2-901/CFTR expression levels, no increase in surface expression was detected using indirect immunofluorescence. Immunoelectron microscopy of unstimulated and FSK-stimulated cells confirmed the M2-901/CFTR redistribution to the PM upon FSK stimulation and demonstrates that the apically inserted M2-901/CFTR originates from a population of subapical vesicles. Our observations may reconcile previous conflicting reports regarding the effect of cAMP stimulation on CFTR trafficking.

  18. Irrigant flow during photon-induced photoacoustic streaming (PIPS) using Particle Image Velocimetry (PIV).

    PubMed

    Koch, Jon D; Jaramillo, David E; DiVito, Enrico; Peters, Ove A

    2016-03-01

    This study aimed to compare fluid movements generated from photon-induced photoacoustic streaming (PIPS) and passive ultrasonic irrigation (PUI). Particle Image Velocimetry (PIV) was performed using 6-μm melamine spheres in water. Measurement areas were 3-mm-long sections of the canal in the coronal, midroot and apical regions for PIPS (erbium/yttrium-aluminium garnet (Er:YAG) laser set at 15 Hz with 20 mJ), or passive ultrasonic irrigation (PUI, non-cutting insert at 30% unit power) was performed in simulated root canals prepared to an apical size #30/0.04 taper. Fluid movement was analysed directly subjacent to the apical ends of ultrasonic insert or fiber optic tips as well as at midroot and apically. During PUI, measured average velocities were around 0.03 m/s in the immediate vicinity of the sides and tip of the ultrasonic file. Speeds decayed to non-measureable values at a distance of about 2 mm from the sides and tip. During PIPS, typical average speeds were about ten times higher than those measured for PUI, and they were measured throughout the length of the canal, at distances up to 20 mm away. PIPS caused higher average fluid speeds when compared to PUI, both close and distant from the instrument. The findings of this study could be relevant to the debriding and disinfecting stage of endodontic therapy. Irrigation enhancement beyond needle irrigation is relevant to more effectively eradicate microorganisms from root canal systems. PIPS may be an alternative approach due to its ability to create high streaming velocities further away from the activation source compared to ultrasonic activation.

  19. Purinergic P2Y receptors in airway epithelia: from ion transport to immune functions.

    PubMed

    Hao, Yuan; Ko, Wing-hung

    2014-02-25

    The regulated transport of salt and water is essential to the integrated function of many organ systems, including the respiratory, reproductive, and digestive tracts. Airway epithelial fluid secretion is a passive process that is driven by osmotic forces, which are generated by ion transport. The main determinant of a luminally-directed osmotic gradient is the mucosal transport of chloride ions (Cl(-)) into the lumen. As with many epithelial cells, a number of classic signal transduction cascades are involved in the regulation of ion transport. There are two well-known intracellular signaling systems: an increase in intracellular Ca(2+) concentration ([Ca(2+)]i) and an increase in the rate of synthesis of cyclic nucleotides, such as cyclic adenosine monophosphate (cAMP). Therefore, Cl(-) secretion is primarily activated via the opening of apical Ca(2+)- or cAMP-dependent Cl(-) channels at the apical membrane. The opening of basolateral Ca(2+)- or cAMP-activated K(+) channels, which hyperpolarizes the cell to maintain the driving force for Cl(-) exit through apical Cl(-) channels that are constitutively open, is also important in regulating transepithelial ion transport. P2Y receptors are expressed in the apical and/or basolateral membranes of virtually all polarized epithelia to control the transport of fluid and electrolytes. Human airway epithelial cells express multiple nucleotide receptors. Extracellular nucleotides, such as UTP and ATP, are calcium-mobilizing secretagogues. They are released into the extracellular space from airway epithelial cells and act on the same cell in an autocrine fashion to stimulate transepithelial ion transport. In addition, recent data support the role of P2Y receptors in releasing inflammatory cytokines in the bronchial epithelium and other immune cells.

  20. A quantitative and qualitative analysis of ultrasonic versus sonic endodontic systems on canal cleanliness and obturation.

    PubMed

    Kanter, Valerie; Weldon, Emily; Nair, Uma; Varella, Claudio; Kanter, Keith; Anusavice, Kenneth; Pileggi, Roberta

    2011-12-01

    The purpose of this study was to compare 2 irrigation techniques by evaluating canal cleanliness and obturation of lateral/accessory canals. Seventy-five extracted canines were instrumented to a size #40/0.06 taper. The EndoActivator (EA) was compared with an ultrasonic unit for final irrigation. Each unit was used for 1 minute each with 6.15% NaOCl and 17% EDTA. A control group received syringe irrigation. Thirty teeth were sectioned and evaluated for debris removal and open dentinal tubules at 3/5 mm from the apical foramen with a scanning electron microscope. Forty-five teeth were examined for obturation of lateral canals. The EA was significantly better in removing debris at all levels when compared with other treatment groups (P < .05) and resulted in obturation of significantly more numbers of lateral canals (P < .01.) The EA provided better obturation of lateral and accessory canals and resulted in less remaining debris. Copyright © 2011 Mosby, Inc. All rights reserved.

  1. Facilitation of Endosomal Recycling by an IRG Protein Homolog Maintains Apical Tubule Structure in Caenorhabditis elegans

    PubMed Central

    Grussendorf, Kelly A.; Trezza, Christopher J.; Salem, Alexander T.; Al-Hashimi, Hikmat; Mattingly, Brendan C.; Kampmeyer, Drew E.; Khan, Liakot A.; Hall, David H.; Göbel, Verena; Ackley, Brian D.; Buechner, Matthew

    2016-01-01

    Determination of luminal diameter is critical to the function of small single-celled tubes. A series of EXC proteins, including EXC-1, prevent swelling of the tubular excretory canals in Caenorhabditis elegans. In this study, cloning of exc-1 reveals it to encode a homolog of mammalian IRG proteins, which play roles in immune response and autophagy and are associated with Crohn’s disease. Mutants in exc-1 accumulate early endosomes, lack recycling endosomes, and exhibit abnormal apical cytoskeletal structure in regions of enlarged tubules. EXC-1 interacts genetically with two other EXC proteins that also affect endosomal trafficking. In yeast two-hybrid assays, wild-type and putative constitutively active EXC-1 binds to the LIM-domain protein EXC-9, whose homolog, cysteine-rich intestinal protein, is enriched in mammalian intestine. These results suggest a model for IRG function in forming and maintaining apical tubule structure via regulation of endosomal recycling. PMID:27334269

  2. The PDZ domain of the guanine nucleotide exchange factor PDZGEF directs binding to phosphatidic acid during brush border formation.

    PubMed

    Consonni, Sarah V; Brouwer, Patricia M; van Slobbe, Eleonora S; Bos, Johannes L

    2014-01-01

    PDZGEF is a guanine nucleotide exchange factor for the small G protein Rap. It was recently found that PDZGEF contributes to establishment of intestinal epithelial polarity downstream of the kinase Lkb1. By binding to phosphatidic acid enriched at the apical membrane, PDZGEF locally activates Rap2a resulting in induction of brush border formation via a pathway that includes the polarity players TNIK, Mst4 and Ezrin. Here we show that the PDZ domain of PDZGEF is essential and sufficient for targeting PDZGEF to the apical membrane of polarized intestinal epithelial cells. Inhibition of PLD and consequently production of phosphatidic acid inhibitis targeting of PDZGEF to the plasma membrane. Furthermore, localization requires specific positively charged residues within the PDZ domain. We conclude that local accumulation of PDZGEF at the apical membrane during establishment of epithelial polarity is mediated by electrostatic interactions between positively charged side chains in the PDZ domain and negatively charged phosphatidic acid.

  3. The PDZ Domain of the Guanine Nucleotide Exchange Factor PDZGEF Directs Binding to Phosphatidic Acid during Brush Border Formation

    PubMed Central

    Consonni, Sarah V.; Brouwer, Patricia M.; van Slobbe, Eleonora S.; Bos, Johannes L.

    2014-01-01

    PDZGEF is a guanine nucleotide exchange factor for the small G protein Rap. It was recently found that PDZGEF contributes to establishment of intestinal epithelial polarity downstream of the kinase Lkb1. By binding to phosphatidic acid enriched at the apical membrane, PDZGEF locally activates Rap2a resulting in induction of brush border formation via a pathway that includes the polarity players TNIK, Mst4 and Ezrin. Here we show that the PDZ domain of PDZGEF is essential and sufficient for targeting PDZGEF to the apical membrane of polarized intestinal epithelial cells. Inhibition of PLD and consequently production of phosphatidic acid inhibitis targeting of PDZGEF to the plasma membrane. Furthermore, localization requires specific positively charged residues within the PDZ domain. We conclude that local accumulation of PDZGEF at the apical membrane during establishment of epithelial polarity is mediated by electrostatic interactions between positively charged side chains in the PDZ domain and negatively charged phosphatidic acid. PMID:24858808

  4. Cyclin D2 in the basal process of neural progenitors is linked to non-equivalent cell fates

    PubMed Central

    Tsunekawa, Yuji; Britto, Joanne M; Takahashi, Masanori; Polleux, Franck; Tan, Seong-Seng; Osumi, Noriko

    2012-01-01

    Asymmetric cell division plays an indispensable role during corticogenesis for producing new neurons while maintaining a self-renewing pool of apical progenitors. The cellular and molecular determinants favouring asymmetric division are not completely understood. Here, we identify a novel mechanism for generating cellular asymmetry through the active transportation and local translation of Cyclin D2 mRNA in the basal process. This process is regulated by a unique cis-regulatory sequence found in the 3′ untranslated region (3′UTR) of the mRNA. Unequal inheritance of Cyclin D2 protein to the basally positioned daughter cell with the basal process confers renewal of the apical progenitor after asymmetric division. Conversely, depletion of Cyclin D2 in the apically positioned daughter cell results in terminal neuronal differentiation. We demonstrate that Cyclin D2 is also expressed in the developing human cortex within similar domains, thus indicating that its role as a fate determinant is ancient and conserved. PMID:22395070

  5. 78 FR 67022 - Airworthiness Directives; Bell Helicopter Textron, Inc., Helicopters

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-08

    ... Apical Industries, Inc. (Apical) emergency float kit installed under Supplemental Type Certificate (STC... service information identified in this AD, contact Apical Industries, Inc., 2608 Temple Heights Drive..., 206L-4, and 407 helicopters with an Apical emergency float kit installed under STC number SR01535LA...

  6. Proteolytic activities in cortex of apical parts of Vicia faba ssp. minor seedling roots during kinetin-induced programmed cell death.

    PubMed

    Kaźmierczak, Andrzej; Doniak, Magdalena; Kunikowska, Anita

    2017-11-01

    Programmed cell death (PCD) is a crucial process in plant development. In this paper, proteolytically related aspects of kinetin-induced PCD in cortex cells of Vicia faba ssp. minor seedlings were examined using morphological, fluorometric, spectrophotometric, and fluorescence microscopic analyses. Cell viability estimation after 46 μM kinetin treatment of seedling roots showed that the number of dying cortex cells increased with treatment duration, reaching maximum after 72 h. Weight of the apical root segments increased with time and was about 2.5-fold greater after 96 h, while the protein content remained unchanged, compared to the control. The total and cysteine-dependent proteolytic activities fluctuated during 1-96-h treatment, which was not accompanied by the changes in the protein amount, indicating that the absolute protein amounts decreased during kinetin-induced PCD. N-ethylmaleimide (NEM), phenylmethylsulfonyl fluoride (PMSF), and Z-Leu-Leu-Nva-H (MG115), the respective cysteine, serine, and proteasome inhibitors, suppressed kinetin-induced PCD. PMSF significantly decreased serine-dependent proteolytic activities without changing the amount of proteins, unlike NEM and MG115. More pronounced effect of PMSF over NEM indicated that in the root apical segments, the most important proteolytic activity during kinetin-induced PCD was that of serine proteases, while that of cysteine proteases may be important for protein degradation in the last phase of the process. Both NEM and PMSF inhibited apoptotic-like structure formation during kinetin-induced PCD. The level of caspase-3-like activity of β1 proteasome subunit increased after kinetin treatment. Addition of proteasome inhibitor MG-115 reduced the number of dying cells, suggesting that proteasomes might play an important role during kinetin-induced PCD.

  7. Cytosolic glyceraldehyde-3-phosphate dehydrogenases play crucial roles in controlling cold-induced sweetening and apical dominance of potato (Solanum tuberosum L.) tubers.

    PubMed

    Liu, Tengfei; Fang, Hui; Liu, Jun; Reid, Stephen; Hou, Juan; Zhou, Tingting; Tian, Zhendong; Song, Botao; Xie, Conghua

    2017-12-01

    Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an important enzyme that functions in producing energy and supplying intermediates for cellular metabolism. Recent researches indicate that GAPDHs have multiple functions beside glycolysis. However, little information is available for functions of GAPDHs in potato. Here, we identified 4 putative cytosolic GAPDH genes in potato genome and demonstrated that the StGAPC1, StGAPC2, and StGAPC3, which are constitutively expressed in potato tissues and cold inducible in tubers, encode active cytosolic GAPDHs. Cosuppression of these 3 GAPC genes resulted in low tuber GAPDH activity, consequently the accumulation of reducing sugars in cold stored tubers by altering the tuber metabolite pool sizes favoring the sucrose pathway. Furthermore, GAPCs-silenced tubers exhibited a loss of apical dominance dependent on cell death of tuber apical bud meristem (TAB-meristem). It was also confirmed that StGAPC1, StGAPC2, and StGAPC3 interacted with the autophagy-related protein 3 (ATG3), implying that the occurrence of cell death in TAB-meristem could be induced by ATG3 associated events. Collectively, the present research evidences first that the GAPC genes play crucial roles in diverse physiological and developmental processes in potato tubers. © 2017 John Wiley & Sons Ltd.

  8. Lamellipodia-based migrations of larval epithelial cells are required for normal closure of the adult epidermis of Drosophila

    PubMed Central

    Bischoff, Marcus

    2012-01-01

    Cell migrations are an important feature of animal development. They are, furthermore, essential to wound healing and tumour progression. Despite recent progress, it is still mysterious how cell migration is spatially and temporally regulated during morphogenesis and how cell migration is coordinated with other cellular behaviours to shape tissues and organs. The formation of the abdominal epithelium of Drosophila during metamorphosis provides an attractive system to study morphogenesis. Here, the diploid adult histoblasts replace the polyploid larval epithelial cells (LECs). Using in vivo 4D microscopy, I show that, besides apical constriction and apoptosis, the LECs undergo extensive coordinated migrations. The migrations follow a transition from a stationary (epithelial) to a migratory mode. The migratory behaviour is stimulated by autocrine Dpp signalling. Directed apical lamellipodia-like protrusions propel the cells. Initially, planar cell polarity determines the orientation of LEC migration. While LECs are migrating they also constrict apically, and changes in activity of the small GTPase Rho1 can favour one behaviour over the other. This study shows that the LECs play a more active role in morphogenesis than previously thought, with their migrations contributing to abdominal closure. It furthermore provides insights into how the migratory behaviour of cells is regulated during morphogenesis. PMID:22230614

  9. Apico-basal forces exerted by apoptotic cells drive epithelium folding.

    PubMed

    Monier, Bruno; Gettings, Melanie; Gay, Guillaume; Mangeat, Thomas; Schott, Sonia; Guarner, Ana; Suzanne, Magali

    2015-02-12

    Epithelium folding is a basic morphogenetic event that is essential in transforming simple two-dimensional epithelial sheets into three-dimensional structures in both vertebrates and invertebrates. Folding has been shown to rely on apical constriction. The resulting cell-shape changes depend either on adherens junction basal shift or on a redistribution of myosin II, which could be driven by mechanical signals. Yet the initial cellular mechanisms that trigger and coordinate cell remodelling remain largely unknown. Here we unravel the active role of apoptotic cells in initiating morphogenesis, thus revealing a novel mechanism of epithelium folding. We show that, in a live developing tissue, apoptotic cells exert a transient pulling force upon the apical surface of the epithelium through a highly dynamic apico-basal myosin II cable. The apoptotic cells then induce a non-autonomous increase in tissue tension together with cortical myosin II apical stabilization in the surrounding tissue, eventually resulting in epithelium folding. Together our results, supported by a theoretical biophysical three-dimensional model, identify an apoptotic myosin-II-dependent signal as the initial signal leading to cell reorganization and tissue folding. This work further reveals that, far from being passively eliminated as generally assumed (for example, during digit individualization), apoptotic cells actively influence their surroundings and trigger tissue remodelling through regulation of tissue tension.

  10. SHORT-ROOT regulates vascular patterning, but not apical meristematic activity in the Arabidopsis root through cytokinin homeostasis

    PubMed Central

    Hao, Yueling; Cui, Hongchang

    2012-01-01

    SHORT-ROOT (SHR) is a key regulator of radial patterning and stem-cell renewal in the Arabidopsis root. Although SHR is expressed in the stele, its function in the vascular tissue was not recognized until recently. In shr, the protoxylem is missing due to the loss of expression of microRNA165A (miR165A) and microRNA166B (miR165B). shr is also defective in lateral root formation, but the mechanism remains unclear. To dissect the SHR developmental pathway, we recently have identified its direct targets at the genome scale by chromatin immunoprecipitation followed by microarray analysis (ChIP-chip). In further studies, we have shown that SHR regulates cytokinin homeostasis through cytokinin oxidase 3 and that this role of SHR is critical to vascular patterning in the root. In this communication we report that SHR also regulates miR165A and miR166B indirectly through its effect on cytokinin homeostasis. Although cytokinin is inhibitory to root growth, the root-apical-meristem defect in shr was not alleviated by reduction of endogenous cytokinin. These results together suggest that SHR regulates vascular patterning, but not root apical meristematic activity, through cytokinin homeostasis. PMID:22476466

  11. An auxin responsive CLE gene regulates shoot apical meristem development in Arabidopsis

    PubMed Central

    Guo, Hongyan; Zhang, Wei; Tian, Hainan; Zheng, Kaijie; Dai, Xuemei; Liu, Shanda; Hu, Qingnan; Wang, Xianling; Liu, Bao; Wang, Shucai

    2015-01-01

    Plant hormone auxin regulates most, if not all aspects of plant growth and development, including lateral root formation, organ pattering, apical dominance, and tropisms. Peptide hormones are peptides with hormone activities. Some of the functions of peptide hormones in regulating plant growth and development are similar to that of auxin, however, the relationship between auxin and peptide hormones remains largely unknown. Here we report the identification of OsCLE48, a rice (Oryza sativa) CLE (CLAVATA3/ENDOSPERM SURROUNDING REGION) gene, as an auxin response gene, and the functional characterization of OsCLE48 in Arabidopsis and rice. OsCLE48 encodes a CLE peptide hormone that is similar to Arabidopsis CLEs. RT-PCR analysis showed that OsCLE48 was induced by exogenously application of IAA (indole-3-acetic acid), a naturally occurred auxin. Expression of integrated OsCLE48p:GUS reporter gene in transgenic Arabidopsis plants was also induced by exogenously IAA treatment. These results indicate that OsCLE48 is an auxin responsive gene. Histochemical staining showed that GUS activity was detected in all the tissue and organs of the OsCLE48p:GUS transgenic Arabidopsis plants. Expression of OsCLE48 under the control of the 35S promoter in Arabidopsis inhibited shoot apical meristem development. Expression of OsCLE48 under the control of the CLV3 native regulatory elements almost completely complemented clv3-2 mutant phenotypes, suggesting that OsCLE48 is functionally similar to CLV3. On the other hand, expression of OsCLE48 under the control of the 35S promoter in Arabidopsis has little, if any effects on root apical meristem development, and transgenic rice plants overexpressing OsCLE48 are morphologically indistinguishable from wild type plants, suggesting that the functions of some CLE peptides may not be fully conserved in Arabidopsis and rice. Taken together, our results showed that OsCLE48 is an auxin responsive peptide hormone gene, and it regulates shoot apical meristem development when expressed in Arabidopsis. PMID:25983737

  12. Periapical cemento-osseous dysplasia: clinicopathological features.

    PubMed

    Roghi, Marco; Scapparone, Chiara; Crippa, Rolando; Silvestrini-Biavati, Armando; Angiero, Francesca

    2014-05-01

    Periapical cemento-osseous dysplasia (PCOD) is a rare benign lesion, often asymptomatic, in which fibrous tissue replaces the normal bone tissue, with metaplasic bone and neo-formed cement. We present a rare case of mandibular PCOD in a woman of 55 years, who presented with moderate swelling and mobility of teeth 32-33-34. Endoral radiography showed that these teeth had been devitalized; they had deep periodontal pockets and marked radicular radiotransparency; the root apices exhibited mixed radiotransparency and radio-opacity. Clinical and radiographical findings led to a diagnosis of periapical rarefying osteitis, and the three teeth were thus extracted. Due to the persistence of swelling and slight pain post-extraction, a cone-beam computed tomographic scan was taken; this showed a mixed radiotransparent and radio-opaque lesion in the area of the extracted teeth. A bone biopsy of the affected area was taken for histopathological evaluation; a diagnosis of PCOD was rendered. This case demonstrates the importance of a full investigation when a patient presents after tooth extraction with non-healing socket, pain, and swelling. A multidisciplinary approach is required to manage these rare cases.

  13. Current limitations and a path forward to improve testing for the environmental assessment of endocrine active substances-presentation

    EPA Science Inventory

    To assess the hazards and risks of possible endocrine active chemicals (EACs), there is a need for robust, validated test methods that detect perturbations of endocrine pathways and provide reliable information for evaluating potential adverse effects on apical endpoints. One iss...

  14. Class III HD-Zip activity coordinates leaf development in Physcomitrella patens.

    PubMed

    Yip, Hoichong Karen; Floyd, Sandra K; Sakakibara, Keiko; Bowman, John L

    2016-11-01

    Land plant bodies develop from meristems, groups of pluripotent stem cells, which may persist throughout the life of a plant or, alternatively, have a transitory existence. Early diverging land plants exhibit indeterminate (persistent) growth in their haploid gametophytic generation, whereas later diverging lineages exhibit indeterminate growth in their diploid sporophytic generation, raising the question of whether genetic machinery directing meristematic functions was co-opted between generations. Class III HD-Zip (C3HDZ) genes are required for the establishment and maintenance of shoot apical meristems in flowering plants. We demonstrate that in the moss Physcomitrella patens, C3HDZ genes are expressed in transitory meristems in both the gametophytic and sporophytic generations, but not in the persistent shoot meristem of the gametyphyte. Loss-of-function of P. patens C3HDZ was engineered using ectopic expression of miR166, an endogenous regulator of C3HDZ gene activity. Loss of C3HDZ gene function impaired the function of gametophytic transitory meristematic activity but did not compromise the functioning of the persistent shoot apical meristem during the gametophyte generation. These results argue against a wholesale co-option of meristematic gene regulatory networks from the gametophyte to the sporophyte during land plant evolution, instead suggesting that persistent meristems with a single apical cell in P. patens and persistent complex meristems in flowering plants are regulated by different genetic programs. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. The role of bile salt export pump mutations in progressive familial intrahepatic cholestasis type II

    PubMed Central

    Wang, Lin; Soroka, Carol J.; Boyer, James L.

    2002-01-01

    PFIC II is a subtype of progressive familial intrahepatic cholestasis (PFIC) that is associated with mutations in the ABCB11 gene encoding the bile salt export pump (BSEP). However it is not known how these mutations cause this disease. To evaluate these mechanisms, we introduced seven PFIC II–associated missense mutations into rat Bsep and assessed their effects on Bsep membrane localization and transport function in MDCK and Sf9 cells, respectively. Five mutations, G238V, E297G, G982R, R1153C, and R1268Q, prevented the protein from trafficking to the apical membrane, and E297G, G982R, R1153C, and R1268Q also abolished taurocholate transport activity, possibly by causing Bsep to misfold. Mutation C336S affected neither Bsep transport activity nor the apical trafficking of Bsep, suggesting that this mutation alone may not cause this disease. D482G did not affect the apical expression but partially decreased the transport activity of Bsep. Mutant G238V was rapidly degraded in both MDCK and Sf9 cells, and proteasome inhibitor resulted in intracellular accumulation of this and other mutants, suggesting proteasome-mediated degradation plays an important role in expression of these PFIC II mutants. Our studies highlight the heterogeneous nature of PFIC II mutations and illustrate the significance of these mutations in the function and expression of Bsep. PMID:12370274

  16. Effect of gravity on apical dominance in Pharbitis nil.

    PubMed

    Kitazawa, Daisuke; Fujii, Nobuharu; Suge, Hiroshi; Takahashi, Hideyuki

    2003-10-01

    When the upper part of main shoot of morning glory (Pharbitis nil) is gently bent down, lateral bud on the bending region is released from apical dominance and starts to elongate. But, clinorotating the bending shoots prevents the release of the lateral bud from apical dominance. These results suggest that gravity affects apical dominance in morning glory. Here we verified the gravity-regulated apical dominance by using a weeping morning glory defective in gravitropic response due to abnormal differentiation of endodermis. That is, bending main shoot of the weeping morning glory hardly caused the lateral bud to elongate. In addition, decapitation of apical bud released the lateral bud from apical dominance, and exogenous auxin applied to the cut surface of the decapitated stem was inhibitory to the outgrowth of the lateral bud in the wild type. However, the effect of auxin was much less in the weeping morning glory. Thus, apical dominance of the weeping morning glory was weaker and less influenced by gravity than that of the wild type, which could occur due to abnormal differentiation of endodermis required for graviperception.

  17. The fast-recycling receptor Megalin defines the apical recycling pathway of epithelial cells

    PubMed Central

    Perez Bay, Andres E.; Schreiner, Ryan; Benedicto, Ignacio; Paz Marzolo, Maria; Banfelder, Jason; Weinstein, Alan M.; Rodriguez-Boulan, Enrique J.

    2016-01-01

    The basolateral recycling and transcytotic pathways of epithelial cells were previously defined using markers such as transferrin (TfR) and polymeric IgA (pIgR) receptors. In contrast, our knowledge of the apical recycling pathway remains fragmentary. Here we utilize quantitative live-imaging and mathematical modelling to outline the recycling pathway of Megalin (LRP-2), an apical receptor with key developmental and renal functions, in MDCK cells. We show that, like TfR, Megalin is a long-lived and fast-recycling receptor. Megalin enters polarized MDCK cells through segregated apical sorting endosomes and subsequently intersects the TfR and pIgR pathways at a perinuclear Rab11-negative compartment termed common recycling endosomes (CRE). Whereas TfR recycles to the basolateral membrane from CRE, Megalin, like pIgR, traffics to subapical Rab11-positive apical recycling endosomes (ARE) and reaches the apical membrane in a microtubule- and Rab11-dependent manner. Hence, Megalin defines the apical recycling pathway of epithelia, with CRE as its apical sorting station. PMID:27180806

  18. Dlg5 maintains apical polarity by promoting membrane localization of Crumbs during Drosophila oogenesis

    PubMed Central

    Luo, Jun; Wang, Heng; Kang, Di; Guo, Xuan; Wan, Ping; Wang, Dou; Chen, Jiong

    2016-01-01

    Apical-basal polarity plays critical roles in the functions of epithelial tissues. However, the mechanisms of epithelial polarity establishment and maintenance remain to be fully elucidated. Here we show that the membrane-associated guanylate kinase (MAGUK) family protein Dlg5 is required for the maintenance of apical polarity of follicle epithelium during Drosophila oogenesis. Dlg5 localizes at the apical membrane and adherens junction (AJ) of follicle epithelium in early stage egg chambers. Specifically, we demonstrate that the major function of Dlg5 is to promote apical membrane localization of Crumbs, since overexpression of Crumbs but not other major apical or AJ components could rescue epithelial polarity defects resulted from loss of Dlg5. Furthermore, we performed a structure-function analysis of Dlg5 and found that the C-terminal PDZ3 and PDZ4 domains are required for all Dlg5’s functions as well as its ability to localize to apical membrane. The N-terminal coiled-coil motif could be individually targeted to the apical membrane, while the central linker region could be targeted to AJ. Lastly, the MAGUK core domains of PDZ4-SH3-GUK could be individually targeted to apical, AJ and basolateral membranes. PMID:27211898

  19. Arp2/3 promotes junction formation and maintenance in the Caenorhabditis elegans intestine by regulating membrane association of apical proteins

    PubMed Central

    Bernadskaya, Yelena Y.; Patel, Falshruti B.; Hsu, Hsiao-Ting; Soto, Martha C.

    2011-01-01

    It has been proposed that Arp2/3, which promotes nucleation of branched actin, is needed for epithelial junction initiation but is less important as junctions mature. We focus here on how Arp2/3 contributes to the Caenorhabditis elegans intestinal epithelium and find important roles for Arp2/3 in the maturation and maintenance of junctions in embryos and adults. Electron microscope studies show that embryos depleted of Arp2/3 form apical actin-rich microvilli and electron-dense apical junctions. However, whereas apical/basal polarity initiates, apical maturation is defective, including decreased apical F-actin enrichment, aberrant lumen morphology, and reduced accumulation of some apical junctional proteins, including DLG-1. Depletion of Arp2/3 in adult animals leads to similar intestinal defects. The DLG-1/AJM-1 apical junction proteins, and the ezrin–radixin–moesin homologue ERM-1, a protein that connects F-actin to membranes, are required along with Arp2/3 for apical F-actin enrichment in embryos, whereas cadherin junction proteins are not. Arp2/3 affects the subcellular distribution of DLG-1 and ERM-1. Loss of Arp2/3 shifts both ERM-1 and DLG-1 from pellet fractions to supernatant fractions, suggesting a role for Arp2/3 in the distribution of membrane-associated proteins. Thus, Arp2/3 is required as junctions mature to maintain apical proteins associated with the correct membranes. PMID:21697505

  20. An Innovative Regenerative Endodontic Procedure Using Leukocyte and Platelet-rich Fibrin Associated with Apical Surgery: A Case Report.

    PubMed

    Pinto, Nelson; Harnish, Alexandra; Cabrera, Carolina; Andrade, Catherine; Druttman, Tony; Brizuela, Claudia

    2017-11-01

    Regenerative endodontic procedures (REPs) associated with apical surgery could represent an alternative treatment strategy for patients whose teeth present incomplete root formation and extensive apical lesions. Leukocyte platelet-rich fibrin (L-PRF) has potential benefits in REPs; it could promote apical root formation and optimal bone healing. The aim of this case report was to describe innovative regenerative endodontic therapy using L-PRF in the root canal and an extensive apical lesion in an immature tooth with dens invaginatus and asymptomatic apical periodontitis. A healthy 20-year-old woman was referred to the dental clinic of the Universidad de Los Andes, Santiago, Chile, for endodontic treatment in tooth # 22 with incomplete root development and an extensive apical lesion. The diagnosis was asymptomatic apical periodontitis associated with dens invaginatus type II. The patient was treated with an innovative approach using L-PRF in REPs associated with apical surgery. Follow-ups were performed at 6 months and 1 year later. They included periapical radiographs, cone-beam computed tomographic imaging, sensitivity, and vitality tests. The clinical evaluations performed at 6 months and 1 year revealed an absence of symptoms. The radiographic evaluations showed that the apical lesion was resolved. The cone-beam images indicated that the root length increased and the walls had thickened. The sensitivity tests were positive, and the laser Doppler flowmetry showed positive blood flow after 1 year. The success of the results in this case report indicate that L-PRF can be used as a complement in apical surgery and REPs and could provide an innovative alternative treatment strategy for complex clinical cases like these. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  1. Live Imaging at the Onset of Cortical Neurogenesis Reveals Differential Appearance of the Neuronal Phenotype in Apical versus Basal Progenitor Progeny

    PubMed Central

    Attardo, Alessio; Calegari, Federico; Haubensak, Wulf; Wilsch-Bräuninger, Michaela; Huttner, Wieland B.

    2008-01-01

    The neurons of the mammalian brain are generated by progenitors dividing either at the apical surface of the ventricular zone (neuroepithelial and radial glial cells, collectively referred to as apical progenitors) or at its basal side (basal progenitors, also called intermediate progenitors). For apical progenitors, the orientation of the cleavage plane relative to their apical-basal axis is thought to be of critical importance for the fate of the daughter cells. For basal progenitors, the relationship between cell polarity, cleavage plane orientation and the fate of daughter cells is unknown. Here, we have investigated these issues at the very onset of cortical neurogenesis. To directly observe the generation of neurons from apical and basal progenitors, we established a novel transgenic mouse line in which membrane GFP is expressed from the beta-III-tubulin promoter, an early pan-neuronal marker, and crossed this line with a previously described knock-in line in which nuclear GFP is expressed from the Tis21 promoter, a pan-neurogenic progenitor marker. Mitotic Tis21-positive basal progenitors nearly always divided symmetrically, generating two neurons, but, in contrast to symmetrically dividing apical progenitors, lacked apical-basal polarity and showed a nearly randomized cleavage plane orientation. Moreover, the appearance of beta-III-tubulin–driven GFP fluorescence in basal progenitor-derived neurons, in contrast to that in apical progenitor-derived neurons, was so rapid that it suggested the initiation of the neuronal phenotype already in the progenitor. Our observations imply that (i) the loss of apical-basal polarity restricts neuronal progenitors to the symmetric mode of cell division, and that (ii) basal progenitors initiate the expression of neuronal phenotype already before mitosis, in contrast to apical progenitors. PMID:18545663

  2. The effects of healing abutments of different size and anatomic shape placed immediately in extraction sockets on peri-implant hard and soft tissues. A pilot study in foxhound dogs.

    PubMed

    López-López, Patricia J; Mareque-Bueno, Javier; Boquete-Castro, Ana; Aguilar-Salvatierra Raya, Antonio; Martínez-González, José M; Calvo-Guirado, José L

    2016-01-01

    The aim of this animal study was to compare the effects of narrow, concave-straight and wide anatomic healing abutments on changes to soft tissues and crestal bone levels around implants immediately placed into extraction sockets in foxhound dogs. Forty-eight titanium implants (Bredent Medical GMBH, Germany) of the same dimensions were placed in six foxhound dogs. They were divided into two groups (n = 24): test (implants with anatomic abutment) and control (implants with concave-straight abutment). The implants were inserted randomly in the post extraction sockets of P2 , P3 , P4, and M1 bilaterally in six dogs. After eight and twelve weeks, the animals were sacrificed and samples extracted containing the implants and the surrounding soft and hard tissues. Soft tissue and crestal bone loss (CBL) were evaluated by histology and histomorphometry. All implants were clinically and histologically osseointegrated. Healing patterns were examined microscopically at eight and twelve weeks. After eight and twelve weeks, for hard tissues, the distance from the implant shoulder to the first bone-to-implant contact (IS-C) was higher for control group in the lingual aspect with statistical significance (P < 0.05). For soft tissues (STL), the distance from the top of the peri-implant mucosa to the apical portion of the junction epithelium (PM-Je) was significantly less on the lingual aspect in the test group (with wider abutment) at eight and twelve weeks (P < 0.05). The distance from the top of the apical portion of the junction epithelium to the first bone-to-implant contact (Je-C) was significantly higher in the test group (wider abutment) in the lingual aspect at eight and twelve weeks (P < 0.05). There was no connective tissue contact with any abutment surface. Within the limitations of this animal study, anatomic healing abutments protect soft and hard tissues and reduce crestal bone resorption compared with concave-straight healing abutments. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  3. [Pathogenesis of apical periodontitis and its effects on the body].

    PubMed

    Márton, I; Bágyi, K; Radics, T; Kiss, C

    1998-01-01

    During the last 25 years there have been major advances in understanding the etiology, pathogenesis and maintenance of inflammatory processes taking place in the periapical space. Polymicrobial infection of the pulp chamber is of primary importance in initiating periapical inflammation. Egress of bacteria and their antigenes stimulate the immune system to form a granulation tissue around the apical area. Local immune response eliminates excess number of invading organisms. However, in parallel with protective reactions, local activity of immunocompetent cells and their soluble products also contribute to tissue damage, bone resorption and perpetuation of inflammation. Present data indicate that interaction of T-lymphocytes and macrophages is crucial in this process.

  4. Apical-access-related complications associated with trans-catheter aortic valve implantation.

    PubMed

    Bleiziffer, Sabine; Piazza, Nicolo; Mazzitelli, Domenico; Opitz, Anke; Bauernschmitt, Robert; Lange, Rüdiger

    2011-08-01

    The left-ventricular trans-apical access has become well established for trans-catheter aortic valve implantation, especially for patients in whom a retrograde trans-arterial implantation is contraindicated. We report on the short- and long-term implications of the apical-access-site-specific complications. Between June 2007 and August 2010, 143 patients were scheduled for trans-apical aortic valve implantation (mean age 80 ± 6 years, n=116 females, mean logistic EuroSCORE (European System for Cardiac Operative Risk Evaluation) 21 ± 13%). The patients are followed up at 30 days, 6 months, and then annually. Severe apical bleeding complications occurred in 10 patients (7%). In three of these patients, the procedure was terminated, and no valve was implanted. In the remaining, the bleeding was controlled with cardiopulmonary bypass support (n=3), via median sternotomy (n=1), or both (n=1) ± later re-exploration. Two additional patients required postprocedural re-exploration for apical bleeding. An apical pseudo-aneurysm developed in two patients (2%), one of whom was treated by surgical revision. Survival was significantly impaired when either apical bleeding, aneurysm, or re-exploration occurred (75% ± 0.082 survival at 30 days and 59% ± 0.122 at 1 year vs 94% ± 0.023 and 80% ± 0.043 in patients without apical complications, p=0.012). Twelve patients (8%) experienced secondary wound healing. An apical hypo- or akinesia was detected in 18/54 (33%) patients at 6 months' echocardiographic investigation, and in 11/30 (37%) 1 year after the procedure. The trans-apical access for trans-catheter aortic valve implantation might be challenging in elderly patients with fragile tissue. Severe bleeding complications or aneurysm formation significantly impairs survival. The clinical impact of subsequent apical hypo- or akinesia has to be further followed up. Copyright © 2011 European Association for Cardio-Thoracic Surgery. Published by Elsevier B.V. All rights reserved.

  5. Altered expression of junctional adhesion molecule 4 in injured podocytes.

    PubMed

    Harita, Yutaka; Miyauchi, Naoko; Karasawa, Tamaki; Suzuki, Koichi; Han, Gi Dong; Koike, Hiroko; Igarashi, Takashi; Shimizu, Fujio; Kawachi, Hiroshi

    2006-02-01

    Recent investigations have revealed the importance of glomerular podocytes with its diaphragm as the major filtration barrier. Junctional adhesion molecule 4 (JAM4) has been identified as a protein that interacts with membrane-associated guanyl kinase inverted (MAGI)-1 and is reported to be expressed on podocytes. To elucidate the role of JAM4 on podocytes, we examined the expression of JAM4 and MAGI-1 in normal and two different proteinuric rat models: puromycin aminonucleoside (PAN) nephropathy and anti-nephrin antibody-induced (ANA) nephropathy, one model with and one without effacement of podocyte foot processes. JAM4 was detected by immunomicroscopy at the apical membrane of normal podocytes. JAM4 immunostaining was focally increased in the podocytes in PAN nephropathy but not in ANA nephropathy. In proteinuric podocytes, the expression of JAM4 was distinct from that of MAGI-1 or other slit diaphragm molecules such as nephrin and ZO-1. Close colocalization of JAM4 and ezrin was maintained in PAN nephropathy. By immunoelectron microscopy, the signals for JAM4 were detected at the free apical membrane of the podocytes with effaced foot processes. Studies with selective detergent extract revealed that the subcellular localization of JAM4 was altered in PAN nephropathy. Thus the altered expression of JAM4 appears to be associated with morphological changes in podocytes and can be a useful marker of injured podocytes. JAM4 may have a different role at the apical membrane besides the role as a junctional molecule and is likely associated with the unique structure of this epithelium.

  6. Microbial community in persistent apical periodontitis: a 16S rRNA gene clone library analysis.

    PubMed

    Zakaria, M N; Takeshita, T; Shibata, Y; Maeda, H; Wada, N; Akamine, A; Yamashita, Y

    2015-08-01

    To characterize the microbial composition of persistent periapical lesions of root filled teeth using a molecular genetics approach. Apical lesion samples were collected from 12 patients (23-80 years old) who visited the Kyushu University Hospital for apicectomy with persistent periapical lesions associated with root filled teeth. DNA was directly extracted from each sample and the microbial composition was comprehensively analysed using clone library analysis of the 16S rRNA gene. Enterococcus faecalis, Candida albicans and specific fimA genotypes of Porphyromonas gingivalis were confirmed using polymerase chain reaction (PCR) analysis with specific primers. Bacteria were detected in all samples, and the dominant findings were P. gingivalis (19.9%), Fusobacterium nucleatum (11.2%) and Propionibacterium acnes (9%). Bacterial diversity was greater in symptomatic lesions than in asymptomatic ones. In addition, the following bacteria or bacterial combinations were characteristic to symptomatic lesions: Prevotella spp., Treponema spp., Peptostreptococcaceae sp. HOT-113, Olsenella uli, Slackia exigua, Selemonas infelix, P. gingivalis with type IV fimA, and a combination of P. gingivalis, F. nucleatum, and Peptostreptococcaceae sp. HOT-113 and predominance of Streptococcus spp. On the other hand, neither Enterococcus faecalis nor C. albicans were detected in any of the samples. Whilst a diverse bacterial species were observed in the persistent apical lesions, some characteristic patterns of bacterial community were found in the symptomatic lesions. The diverse variation of community indicates that bacterial combinations as a community may cause persistent inflammation in periapical tissues rather than specific bacterial species. © 2014 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  7. Maxillary canine morphology: comparative and descriptive analysis from periapical radiographs and cone beam computed tomography.

    PubMed

    Torres, Hianne Miranda de; Arruda, Julyanna Jacinto de; Silva-Filho, João Manoel da; Faria, Danielle Lago Bruno de; Nascimento, Monikelly Carmo Chagas; Torres, Érica Miranda de

    2017-01-01

    The anatomical characteristics of permanent maxillary canines were evaluated through visual examination, periapical radiography, and cone beam computed tomography (CBCT), and measurements obtained from the images and directly on the teeth were compared. Fifty extracted human maxillary canines were classified according to the side of the mouth. The direction of root curvature and location of the apical foramen were also verified. Periapical radiographs and CBCTs of the specimens were obtained. The number of root canals was verified. Tooth length and the mesiodistal and buccopalatal widths of the root were measured directly on the specimens as well as on the radiographs and CBCTs. Data were analyzed by chi-square testing and analysis of variance (α = 0.05). All teeth-26 (52%) from the right side of the dental arch and 24 (48%) from the left-had only 1 main canal each. The apical foramen was located exactly in the root apex in 34 teeth (68%). Root curvature toward the distal side was observed in the apical third in 23 teeth (46%). There were no statistically significant differences between the canines' arch side and either the foramen location (P = 0.104) or the root curvature (P = 0.215). No statistically significant differences were found in measurements of tooth length (P = 0.669), mesiodistal root width (P = 0.517), or buccopalatal root width (P = 0.672) obtained from specimens and images. Both CBCTs and periapical radiographs were reliable for determining the tooth length, mesiodistal root width, and buccopalatal root width of maxillary canines and produced statistically similar measurements.

  8. Root Canal Cleaning Efficacy of Rotary and Hand Files Instrumentation in Primary Molars

    PubMed Central

    Nazari Moghaddam, Kiumars; Mehran, Majid; Farajian Zadeh, Hamideh

    2009-01-01

    INTRODUCTION: Pulpectomy of primary teeth is commonly carried out with hand files and broaches; a tricky and time consuming procedure. The purpose of this in vitro study was to compare the cleaning efficacy and time taken for instrumentation of deciduous molars using hand K-files and Flex Master rotary system. MATERIALS AND METHODS: In this study, 68 canals of 23 extracted primary molars with at least two third intact roots and 7-12 mm length were selected. After preparing an access cavity, K-file size #15 was introduced into the root canal and India ink was injected with an insulin syringe. Sixty samples were randomly divided in to experimental groups in group I (n=30), root canals were prepared with hand K-files; in group II (n=30), rotary Flex Master files were used for instrumentation, and in group III 8 remained samples were considered as negative controls. After clearing and root sectioning, the removal of India ink from cervical, middle, and apical thirds was scored. Data was analyzed using student's T-test and Mann-Whitney U test. RESULTS: There was no significant difference between experimental groups cleaning efficacy at the cervical, middle and apical root canal thirds. Only the coronal third scored higher in the hand instrumented group (P<0.001). Instrumentation with Flex Master rotary files was significantly less time consuming (P<0.001). CONCLUSION: Although there was no difference in cleanliness efficacy at the apical and middle thirds, the coronal third was more effectively cleaned with hand files. Predictably, time efficiency was a significant advantage with rotary technique. PMID:23940486

  9. Apical Extrusion of Debris after Canal Preparation with Hand-Files Used Manually or Installed on Reciprocating Air-Driven Handpiece in Straight and Curved Canals.

    PubMed

    Labbaf, Hossein; Shakeri, Leila; Orduie, Reza; Bastami, Farshid

    2015-01-01

    Apical debris extrusion (DE) subsequent to root canal instrumentation, is one of the most important causes of endodontic flare-ups. The aim of this study was to compare the amount of DE after root canal instrumentation using nickel-titanium (NiTi) hand files with step-back manual technique or installed on reciprocating handpiece. This study was conducted on mesiobuccal (MB) roots of extracted maxillary first molars (n=20) and roots of mandibular premolars (n=20) that were randomly divided into two groups (n=20) according to the armamentarium used for canal preparation (air-driven reciprocating handpiece or hand instrumentation). In each group, the MB and premolar roots were prepared with the main apical sizes of 35 and 40, respectively. The extruded debris were collected and weighed. Finally, the mean dry weights were compared using ANOVA and t-test, and Tukey's Multiple Comparisons Procedures were used to determine the significant differences in amounts of DE. The level of significance was set at 0.05. Regardless of the type of teeth, the mean values of DE, were significantly lower in the handpiece group (P<0.0001). In addition, significantly lower amounts of DE was observed in premolars in similar group (P<0.001). However, this difference was not significant in MB roots of molars (P=0.20). Root canal preparation with reciprocating handpiece can lead to significantly lower debris extrusion than the manual step-back technique. In handpiece-prepared canals, the amount of extruded debris was significantly lower in premolar teeth.

  10. Comparing the Marginal Adaptation of Cold Ceramic and Mineral Trioxide Aggregate by Means of Scanning Electron Microscope: An In vitro Study.

    PubMed

    Mokhtari, Fatemeh; Modaresi, Jalil; Javadi, Gholamreza; Davoudi, Amin; Badrian, Hamid

    2015-09-01

    Long-term success of endodontic surgeries is often influenced by the type of root-end filling material (RFM). The aim of present study was to compare the marginal adaptation of two different RFM, cold ceramic (CC) and mineral trioxide aggregate (MTA), using scanning electron microscope (SEM). About 20 extracted human single-rooted teeth were collected and stored into sodium hypochlorite 5.25%. The teeth were decronated from the cemento-enamel junction to prepare 16 mm roots. The working length was measured, and 1/3 coronal of the canal was prepared by Gates-Glidden drills. Apical flaring was followed by K file size # 40-70 based on step back technique. After filling of the canals, 3 mm above the apex was cut at 90° to the long axis. Furthermore, 3 mm of the filling was removed from the apical part using the ultrasonic device. All of the prepared specimens were divided into two groups and were retro filled by MTA and CC. The roots were cut horizontally from 1 mm above the apical part, and dentin-filling material interface was observed by SEM. Finally, the collected data were analyzed by Mann-Whitney test and using SPSS software version 18 at a significant level of 0.05. The mean interfacial adaptation was higher in CC group. However, no significant differences were observed by statistical test (P = 0.35). Both CC and MTA had similar marginal adaptation as RFM however in vivo studies are recommended for better determination.

  11. Thermal effects from modified endodontic laser tips used in the apical third of root canals with erbium-doped yttrium aluminium garnet and erbium, chromium-doped yttrium scandium gallium garnet lasers.

    PubMed

    George, Roy; Walsh, Laurence J

    2010-04-01

    To evaluate the temperature changes occurring on the apical third of root surfaces when erbium-doped yttrium aluminium garnet (Er:YAG) and erbium, chromium-doped yttrium scandium gallium garnet (Er,Cr:YSGG) laser energy was delivered with a tube etched, laterally emitting conical tip and a conventional bare design optical fiber tip. Thermal effects of root canal laser treatments on periodontal ligament cells and alveolar bone are of concern in terms of safety. A total of 64 single-rooted extracted teeth were prepared 1 mm short of the working length using rotary nickel-titanium Pro-Taper files to an apical size corresponding to a F5 Pro-Taper instrument. A thermocouple located 2 mm from the apex was used to record temperature changes arising from delivery of laser energy through laterally emitting conical tips or plain tips, using an Er:YAG or Er,Cr:YSGG laser. For the Er:YAG and Er,Cr:YSGG systems, conical fibers showed greater lateral emissions (452 + 69% and 443 + 64%) and corresponding lower forward emissions (48 + 5% and 49 + 5%) than conventional plain-fiber tips. All four combinations of laser system and fiber design elicited temperature increases less than 2.5 degrees C during lasing. The use of water irrigation attenuated completely the thermal effects of individual lasing cycles. Laterally emitting conical fiber tips can be used safely under defined conditions for intracanal irradiation without harmful thermal effects on the periodontal apparatus.

  12. Spontaneous release from mossy fiber terminals inhibits Ni2+-sensitive T-type Ca2+ channels of CA3 pyramidal neurons in the rat organotypic hippocampal slice.

    PubMed

    Reid, Christopher A; Xu, Shenghong; Williams, David A

    2008-01-01

    Mossy fibers (axons arising from dentate granule cells) form large synaptic contacts exclusively onto the proximal apical dendrites of CA3 pyramidal neurons. They can generate large synaptic currents that occur in close proximity to the soma. These properties mean that active conductance in the proximal apical dendrite could have a disproportionate influence on CA3 pyramidal neuron excitability. Ni(2+)-sensitive T-type Ca(2+) channels are important modulators of dendritic excitability. Here, we use an optical approach to determine the contribution of Ni(2+) (100 microM)-sensitive Ca(2+) channels to action potential (AP) elicited Ca(2+) flux in the soma, proximal apical and distal apical dendrites. At resting membrane potentials Ni(2+)-sensitive Ca(2+) channels do not contribute to the Ca(2+) signal in the proximal apical dendrite, but do contribute in the other cell regions. Spontaneous release from mossy fiber terminals acting on 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX)-sensitive postsynaptic channels underlies a tonic inhibition of Ni(2+)-sensitive channels. Chelating Zn(2+) with CaEDTA blocks CNQX-sensitive changes in Ca(2+) flux implicating a mechanistic role of this ion in T-type Ca(2+) channel block. To test if this inhibition influenced excitability, progressively larger depolarizing pulses were delivered to CA3 pyramidal neurons. CNQX significantly reduced the size of the depolarizing step required to generate APs and increased the absolute number of APs per depolarizing step. This change in AP firing was completely reversed by the addition of Ni(2+). This mechanism may reduce the impact of T-type Ca(2+) channels in a region where large synaptic events are common.

  13. Removal efficiency of calcium hydroxide dressing from the root canal without chemically active adjuvant.

    PubMed

    Maalouf, Lara; Zogheib, Carla; Naaman, Alfred

    2013-03-01

    Compare the efficiency in removing two calcium hydroxide [Ca(OH)2] preparations from root canal and apical third using single use syringe, ultrasonics and RinsEndo((®)) with saline solution. One hundred and eighty human singlerooted teeth were instrumented using ProTaper((®)) rotary system, divided into two groups according to Ca(OH)2 filling. A: Powder mixed with water, B: Pulpdent((®)). Each group was divided into three subgroups for the irrigation: A1, B1: Single use syringe; A2, B2: Ultrasonics; A3, B3: RinsEndo((®)). Teeth were split longitudinally, photographed and imported into Adobe Photoshop. The percentage ratios of Ca(OH)2 remaining in the canal and in the apical third were calculated. Data were statistically analyzed using 'ANOVA two-way' and 'univariate tests'. (a) Remnants of medicament were found in all teeth, (b) no statistically significant difference in the elimination of both Ca(OH)2 from the entire canal (p = 0.436), however, mixed powder was better eliminated from the apical third (p = 0.005), (c) no statistically significant difference among the irrigation techniques in the whole canal (p = 0.608), though, RinsEndo((®)) and ultrasonics were the most effective in cleaning the apical third (p = 0.032) when mixed powder was used. None of the techniques removed completely Ca(OH)2 from the canal. In the apical third, RinsEndo((®)) and ultrasonics were the most effective when mixed powder was used. Ca(OH)2, the most commonly used intracanal dressing, should be completely eliminated before the obturation to assure a good endodontic sealing. Based on the results of this study, RinsEndo((®)) and ultrasonics were the most effective in removal of Ca(OH)2 especially the powder mixed with water presentation.

  14. The barley MATE gene, HvAACT1, increases citrate efflux and Al3+ tolerance when expressed in wheat and barley

    PubMed Central

    Zhou, Gaofeng; Delhaize, Emmanuel; Zhou, Meixue; Ryan, Peter R.

    2013-01-01

    Background and Aims Aluminium is toxic in acid soils because the soluble Al3+ inhibits root growth. A mechanism of Al3+ tolerance discovered in many plant species involves the release of organic anions from root apices. The Al3+-activated release of citrate from the root apices of Al3+-tolerant genotypes of barley is controlled by a MATE gene named HvAACT1 that encodes a citrate transport protein located on the plasma membrane. The aim of this study was to investigate whether expressing HvAACT1 with a constitutive promoter in barley and wheat can increase citrate efflux and Al3+ tolerance of these important cereal species. Methods HvAACT1 was over-expressed in wheat (Triticum aestivum) and barley (Hordeum vulgare) using the maize ubiquitin promoter. Root apices of transgenic and control lines were analysed for HvAACT1 expression and organic acid efflux. The Al3+ tolerance of transgenic and control lines was assessed in both hydroponic solution and acid soil. Key Results and Conclusions Increased HvAACT1 expression in both cereal species was associated with increased citrate efflux from root apices and enhanced Al3+ tolerance, thus demonstrating that biotechnology can complement traditional breeding practices to increase the Al3+ tolerance of important crop plants. PMID:23798600

  15. Effects of uniform extracellular DC electric fields on excitability in rat hippocampal slices in vitro.

    PubMed

    Bikson, Marom; Inoue, Masashi; Akiyama, Hiroki; Deans, Jackie K; Fox, John E; Miyakawa, Hiroyoshi; Jefferys, John G R

    2004-05-15

    The effects of uniform steady state (DC) extracellular electric fields on neuronal excitability were characterized in rat hippocampal slices using field, intracellular and voltage-sensitive dye recordings. Small electric fields (1 s) changes in neuronal excitability. Electric fields perpendicular to the apical-dendritic axis did not induce somatic polarization, but did modulate orthodromic responses, indicating an effect on afferents. These results demonstrate that DC fields can modulate neuronal excitability in a time-dependent manner, with no clear threshold, as a result of interactions between neuronal compartments, the non-linear properties of the cell membrane, and effects on afferents.

  16. Gross, computed tomographic and histological findings in mandibular cheek teeth extracted from horses with clinical signs of pulpitis due to apical infection.

    PubMed

    Casey, M B; Pearson, G R; Perkins, J D; Tremaine, W H

    2015-09-01

    The most prevalent type of equine dental pulpitis due to apical infection is not associated with coronal fractures or periodontal disease. The pathogenesis of this type of pulpitis is not fully understood. Computed tomography (CT) is increasingly used to investigate equine dental disorders. However, gross, tomographic and histopathological changes in equine dental pulpitis have not been compared previously. To compare gross, CT and histological appearances of sectioned mandibular cheek teeth extracted from horses with clinical signs of pulpitis without coronal fractures or periodontal disease. To contribute to understanding the pathogenesis of equine dental pulpitis. Descriptive study using diseased and healthy teeth. Mandibular cheek teeth extracted from horses with clinical signs of pulpitis (cases), and from cadavers with no history of dental disease (controls), were compared using CT in the transverse plane at 1 mm intervals. Teeth were then sectioned transversely, photographed and processed for histopathological examination. Tomographs were compared with corresponding gross and histological sections. Cement, dentine and bone had similar ranges of attenuation (550-2000 Hounsfield Units, HU) in tomographs but could be differentiated from pulp (-400 to 500 HU) and enamel (> 2500 HU). Twelve discrete dental lesions were identified grossly, 10 of which were characterised histologically. Reactive and reparative dentinogenesis and extensive pulpar mineralisation, previously undescribed, were identified. Pulpar oedema, neutrophilic inflammation, cement and enamel defects, and reactive cemental deposition were also observed. The CT and pathological findings corresponded well where there was mineralised tissue deposited, defects in mineralised tissue, or food material in the pulpar area. Pulpar and dentinal necrosis and cement destruction, evident grossly and histologically, did not correspond to CT changes. Computed tomography is useful for identifying deposition and defects of mineralised material but less useful for identifying inflammation and tissue destruction. The equine dentine-pulp complex responds to insult with reactive and reparative changes. © 2014 EVJ Ltd.

  17. Vesicular secretion of auxin

    PubMed Central

    Schlicht, Markus; Volkmann, Dieter; Mancuso, Stefano

    2008-01-01

    The plant hormone auxin is secreted in root apices via phospholipase Dζ2 (PLDζ2) activity which produces specific population of phosphatidic acid that stimulates secretion of vesicles enriched with auxin. These vesicles were reported to be localized at plant synapses which are active in auxin secretion, especially at the transition zone of the root apex. There are several implications of this vesicular secretion of auxin. In root apices, auxin emerges as plant neurotransmitter-like signal molecule which coordinates activities of adjacent cells via electric and chemical signaling. Putative quantal release of auxin after electrical stimulation, if confirmed, would be part of neuronal communication between plant cells. As auxin transport across plant synapses is tightly linked with integrated sensory perception of environment, especially of omnipresent gravity and light, this process is proposed to mediate the plant perception of environment. These neuronal features allow sessile plants to integrate multitude of sensory signals into the adaptive behavior of whole plants and the animal-like exploratory behavior of growing roots. PMID:19704646

  18. Decellularized Human Dental Pulp as a Scaffold for Regenerative Endodontics.

    PubMed

    Song, J S; Takimoto, K; Jeon, M; Vadakekalam, J; Ruparel, N B; Diogenes, A

    2017-06-01

    Teeth undergo postnatal organogenesis relatively late in life and only complete full maturation a few years after the crown first erupts in the oral cavity. At this stage, development can be arrested if the tooth organ is damaged by either trauma or caries. Regenerative endodontic procedures (REPs) are a treatment alternative to conventional root canal treatment for immature teeth. These procedures rely on the transfer of apically positioned stem cells, including stem cells of the apical papilla (SCAP), into the root canal system. Although clinical success has been reported for these procedures, the predictability of expected outcomes and the organization of the newly formed tissues are affected by the lack of an available suitable scaffold that mimics the complexity of the dental pulp extracellular matrix (ECM). In this study, we evaluated 3 methods of decellularization of human dental pulp to be used as a potential autograft scaffold. Tooth slices of human healthy extracted third molars were decellularized by 3 different methods. One of the methods generated the maximum observed decellularization with minimal impact on the ECM composition and organization. Furthermore, recellularization of the scaffold supported the proliferation of SCAP throughout the scaffold with differentiation into odontoblast-like cells near the dentinal walls. Thus, this study reports that human dental pulp from healthy extracted teeth can be successfully decellularized, and the resulting scaffold supports the proliferation and differentiation of SCAP. The future application of this form of an autograft in REPs can fulfill a yet unmet need for a suitable scaffold, potentially improving clinical outcomes and ultimately promoting the survival and function of teeth with otherwise poor prognosis.

  19. Molecular characterization of the apical organ of the anthozoan Nematostella vectensis

    PubMed Central

    Sinigaglia, Chiara; Busengdal, Henriette; Lerner, Avi; Oliveri, Paola; Rentzsch, Fabian

    2015-01-01

    Apical organs are sensory structures present in many marine invertebrate larvae where they are considered to be involved in their settlement, metamorphosis and locomotion. In bilaterians they are characterised by a tuft of long cilia and receptor cells and they are associated with groups of neurons, but their relatively low morphological complexity and dispersed phylogenetic distribution have left their evolutionary relationship unresolved. Moreover, since apical organs are not present in the standard model organisms, their development and function are not well understood. To provide a foundation for a better understanding of this structure we have characterised the molecular composition of the apical organ of the sea anemone Nematostella vectensis. In a microarray-based comparison of the gene expression profiles of planulae with either a wildtype or an experimentally expanded apical organ, we identified 78 evolutionarily conserved genes, which are predominantly or specifically expressed in the apical organ of Nematostella. This gene set comprises signalling molecules, transcription factors, structural and metabolic genes. The majority of these genes, including several conserved, but previously uncharacterized ones, are potentially involved in different aspects of the development or function of the long cilia of the apical organ. To demonstrate the utility of this gene set for comparative analyses, we further analysed the expression of a subset of previously uncharacterized putative orthologs in sea urchin larvae and detected expression for twelve out of eighteen of them in the apical domain. Our study provides a molecular characterization of the apical organ of Nematostella and represents an informative tool for future studies addressing the development, function and evolutionary history of apical organ cells. PMID:25478911

  20. Constitutive apical membrane recycling in Aplysia enterocytes.

    PubMed

    Keeton, Robert Aaron; Runge, Steven William; Moran, William Michael

    2004-11-01

    In Aplysia californica enterocytes, alanine-stimulated Na+ absorption increases both apical membrane exocytosis and fractional capacitance (fCa; a measure of relative apical membrane surface area). These increases are thought to reduce membrane tension during periods of nutrient absorption that cause the enterocytes to swell osmotically. In the absence of alanine, exocytosis and fCa are constant. These findings imply equal rates of constitutive endocytosis and exocytosis and constitutive recycling of the apical plasma membrane. Thus, the purpose of this study was to confirm and determine the relative extent of constitutive apical membrane recycling in Aplysia enterocytes. Biotinylated lectins are commonly used to label plasma membranes and to investigate plasma membrane recycling. Of fourteen biotinylated lectins tested, biotinylated wheat germ agglutinin (bWGA) bound preferentially to the enterocytes apical surface. Therefore, we used bWGA, avidin D (which binds tightly to biotin), and the UV fluorophore 7-amino-4-methylcoumarin-3-acetic acid (AMCA)-conjugated avidin D to assess the extent of constitutive apical membrane recycling. A temperature-dependent (20 vs. 4 degrees C) experimental protocol employed the use of two tissues from each of five snails and resulted in a approximately 60% difference in apical surface fluorescence intensity. Because the extent of membrane recycling is proportional to the difference in surface fluorescence intensity, this difference reveals a relatively high rate of constitutive apical membrane recycling in Aplysia enterocytes.

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