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Sample records for acid-schiff pas staining

  1. Periodic Acid-Schiff Staining Parallels the Immunoreactivity Seen By Direct Immunofluorescence in Autoimmune Skin Diseases

    PubMed Central

    Abreu Velez, Ana Maria; Upegui Zapata, Yulieth Alexandra; Howard, Michael S

    2016-01-01

    Background: In many countries and laboratories, techniques such as direct immunofluorescence (DIF) are not available for the diagnosis of skin diseases. Thus, these laboratories are limited in the full diagnoses of autoimmune skin diseases, vasculitis, and rheumatologic diseases. In our experience with these diseases and the patient's skin biopsies, we have noted a positive correlation between periodic acid-Schiff (PAS) staining and immunofluorescence patterns; however, these were just empiric observations. In the current study, we aim to confirm these observations, given the concept that the majority of autoantibodies are glycoproteins and should thus be recognized by PAS staining. Aims: To compare direct immunofluorescent and PAS staining, in multiple autoimmune diseases that are known to exhibit specific direct immunofluorescent patterns. Materials and Methods: We studied multiple autoimmune skin diseases: Five cases of bullous pemphigoid, five cases of pemphigus vulgaris, ten cases of cutaneous lupus, ten cases of autoimmune vasculitis, ten cases of lichen planus (LP), and five cases of cutaneous drug reactions (including one case of erythema multiforme). In addition, we utilized 45 normal skin control specimens from plastic surgery reductions. Results: We found a 98% positive correlation between DIF and PAS staining patterns over all the disease samples. Conclusion: We recommend that laboratories without access to DIF always perform PAS staining in addition to hematoxylin and eosin (H&E) staining, for a review of the reactivity pattern. PMID:27114972

  2. Periodic acid-Schiff-positive organisms in primary cutaneous Bacillus cereus infection. Case report and an investigation of the periodic acid-Schiff staining properties of bacteria.

    PubMed

    Khavari, P A; Bolognia, J L; Eisen, R; Edberg, S C; Grimshaw, S C; Shapiro, P E

    1991-04-01

    Primary cutaneous Bacillus cereus infection frequently presents as a single necrotic bulla on the extremity of an immunocompromised patient. In lesional biopsy specimens and smears, the large gram-positive rods of B cereus may be mistaken for Clostridium species. This is a potentially serious error, as Bacillus species are resistant to penicillin and other beta-lactam antibiotics. We studied a case in which large periodic acid-Schiff-staining organisms were seen in the biopsy specimen from a necrotic bulla on the finger of a neutropenic patient with diffuse large cell lymphoma. The tissue biopsy specimen subsequently yielded a pure culture of B cereus. Staining with periodic acid-Schiff was then performed on a series of bacterial species in human tissue and from smears of culture colonies. The following bacterial species were found to be consistently periodic acid-Schiff positive after diastase digestion: B cereus, Corynebacterium diphtheriae, Propionibacterium acnes, Klebsiella pneumoniae, and Micrococcus luteus. PMID:1900984

  3. Orthotopic Human Choroidal Melanoma Xenografts in Nude Rats with Aggressive and Nonaggressive PAS Staining Patterns

    PubMed Central

    Braun, Rod D.; Abbas, Asad

    2007-01-01

    PURPOSE Choroidal melanoma is the most common primary ocular cancer among the adult population. Patient survival has been linked to the periodic acid-Schiff base (PAS)–positive vascular patterns in the tumors. The presence of PAS-positive loops or cross-linking parallel channels is a marker of an aggressive tumor. The purpose of this study was to develop new xenograft models of human choroidal melanoma that predictably demonstrate the PAS staining patterns associated with nonaggressive and aggressive tumors in humans. METHODS Three human choroidal melanoma cell lines (C918, M619, and OCM-1) were used. C918 and M619 are considered aggressive, based on their ability to form PAS-positive channels in vitro. The nonaggressive OCM-1 cells do not form these channels. C918, M619, and OCM-1 spheroids were grown and implanted in the suprachoroidal space of 20, 17, and 16 WAG/RijHs-rnu nude rats, respectively. Tumors were grown for 1 to >4 weeks, and histology was performed to evaluate tumor growth and determine PAS labeling patterns. RESULTS Growth of C918, M619, and OCM-1 xenografts were histologically verified in 20/20, 15/17, and 16/16 rats, respectively. PAS staining revealed loops and cross-linking parallel channels, typical of aggressive tumors in patients, in 90% of C918 and 100% of M619 xenografts. Only 4 of 16 OCM-1 xenografts showed PAS-positive loops. The rest showed no PAS staining or only perivascular staining, indicative of nonaggressive tumors. CONCLUSIONS It is possible to grow human choroidal melanoma orthotopic xenografts in nude rats that reproduce the PAS staining patterns associated with aggressive and nonaggressive choroidal melanomas in patients. PMID:16384938

  4. Periodic acid-Schiff-positive loops and networks as a prognostic factor in oral mucosal melanoma.

    PubMed

    Song, Hao; Jing, Guangping; Wang, Lizhen; Guo, Wei; Ren, Guoxin

    2016-04-01

    The prognostic factors of oral mucosal melanoma (OMM), a rare and aggressive neoplasm, remain to be determined. The aim of this study is to investigate the prognostic significance of vasculogenic mimicry in OMM. The clinical data of 62 patients with primary OMM treated in Shanghai Ninth People's Hospital from April 2007 to April 2012 were retrieved and analyzed retrospectively. Staining of periodic acid-Schiff (PAS) and CD31 immunohistochemistry were performed to evaluate the prognostic value of PAS-positive patterns, blood lakes, and microvascular density. PAS-positive loops and networks (P<0.001) as well as blood lakes (P=0.040) were found to be predictors of overall survival (OS). The presence of PAS-positive loops and networks was an independent prognostic factor of poor OS in multivariate analysis (P=0.002). Although the presence of PAS-positive loops and networks was associated with hematogenous metastasis (P=0.041) and lymphogenous metastasis (P=0.041), it was not an independent predictor of both types of metastasis in multivariate analysis. Microvascular density was not associated with OS (P=0.627) and metastasis of OMM patients. PAS-positive loops and networks have a significant prognostic value in OMM. Detection of PAS-positive patterns may lead to better staging and serve as a prognostic parameter of OMM. PMID:26636907

  5. Comparing The Efficacy of Hematoxylin and Eosin, Periodic Acid Schiff and Fluorescent Periodic Acid Schiff-Acriflavine Techniques for Demonstration of Basement Membrane in Oral Lichen Planus: A Histochemical Study

    PubMed Central

    Pujar, Ashwini; Pereira, Treville; Tamgadge, Avinash; Bhalerao, Sudhir; Tamgadge, Sandhya

    2015-01-01

    Background: Basement membrane (BM) is a thick sheet of extracellular matrix molecules, upon which epithelial cells attach. Various immunohistochemical studies in the past have been carried out but these advanced staining techniques are expensive and not feasible in routine laboratories. Although hematoxylin and eosin (H-E) is very popular among pathologists for looking at biopsies, the method has some limitations. This is where special stains come handy. Aims and Objectives: The aim of the present study was to demonstrate and compare the efficacy of H-E, periodic acid Schiff (PAS) and fluorescent periodic acid–acriflavine staining techniques for the basement membrane and to establish a histochemical stain which could be cost effective, less time consuming, and unambiguous for observation of the basement membrane zone. Materials and Methods: A total number of 40 paraffin-embedded tissue sections of known basement membrane containing tissues including 10 – Normal oral mucosa (NOM) and 30 – oral lichen planus (OLP) were considered in the study. Four-micron-thick sections of each block were cut and stained with H-E stain, PAS and fluorescent periodic acid–acriflavine stain. Sections were evaluated by three oral pathologists independently for continuity, contrast and pattern. Results: Though all the three stains showed favorable features at different levels, acriflavine stain was better than the other stains in demonstrating BM continuity, contrast and also the pattern followed by PAS stain. Acriflavine stain was the better in demonstrating a fibrillar pattern of a BM. Acriflavine stains a BM distinctly and is less time consuming and easy to carry out using readily available dyes as compared to other stains. Conclusion: The continuity and contrast along with the homogenous pattern and the afibrillar pattern of the BM was better demonstrated by acriflavine followed by the PAS stain. PMID:26538690

  6. Prognostic significance of periodic acid-Schiff-positive patterns in primary cutaneous melanoma.

    PubMed

    Warso, M A; Maniotis, A J; Chen, X; Majumdar, D; Patel, M K; Shilkaitis, A; Gupta, T K; Folberg, R

    2001-03-01

    The patterns of periodic acid-Schiff (PAS) staining of extracellular matrix in histological sections of certain melanomas may be predictive of outcome. Recent in vitro and molecular genetic data suggest that the appearance of these patterns in both uveal and cutaneous melanoma is a function of aggressive tumor cells. We studied 96 patients with primary cutaneous melanomas treated at the University of Illinois at Chicago who were monitored for disease-free survival. Survival probabilities were determined by Kaplan-Meier estimates, and prognostic factors were evaluated by multivariate analysis. By univariate analysis, there was a significant decrease in disease-free survival among patients whose tumors contained parallel with cross-linking or network patterns (PXNs; P = 0.0070). Stepwise regression with Cox models that included the combinations of the PAS-positive patterns, tumor thickness, female gender, ulceration, and age yielded a model with thickness and the PAS-positive parallel with cross-linking or networks. Despite the relatively small sample size in this study, the detection of the PAS-positive parallel with cross-linking or networking in cutaneous melanoma was associated with a decrease in disease-free outcome. Additional studies of the prognostic significance of these patterns is warranted on larger data sets. PMID:11297236

  7. Periodic acid-Schiff granules in the brain of aged mice: From amyloid aggregates to degenerative structures containing neo-epitopes.

    PubMed

    Manich, Gemma; Cabezón, Itsaso; Augé, Elisabet; Pelegrí, Carme; Vilaplana, Jordi

    2016-05-01

    Brain ageing in mice leads to the progressive appearance and expansion of degenerative granular structures frequently referred as "PAS granules" because of their positive staining with periodic acid-Schiff (PAS). PAS granules are present mainly in the hippocampus, although they have also been described in other brain areas such as piriform and entorhinal cortices, and have been observed in other mammals than mice, like rats and monkeys. PAS granules have been identified as a wide range of brain deposits related to numerous neurodegenerative diseases, such as amyloid deposits, neurofibrillary tangles, Lafora bodies, corpora amylacea and polyglucosan bodies, and these identifications have generated controversy and particular theories about them. We have recently reported the presence of a neo-epitope in mice hippocampal PAS granules and the existence of natural IgM auto-antibodies directed against the neo-epitope in the plasma of the animals. The significance of the neo-epitope and the autoantibodies is discussed in this review. Moreover, we observed that the IgM anti-neo-epitope is frequently present as a contaminant in numerous commercial antibodies and is responsible of a considerable amount of false positive immunostainings, which may produce misinterpretations in the identification of the granules. Now that this point has been clarified, this article reviews and reconsiders the nature and physiopathological significance of these degenerative granules. Moreover, we suggest that neo-epitopes may turn into a useful brain-ageing biomarker and that autoimmunity could become a new focus in the study of age-related degenerative processes. PMID:26970374

  8. Comparison of Special Stains for Keratin with Routine Hematoxylin and Eosin Stain

    PubMed Central

    Rao, Roopa S; Patil, Shankargouda; Majumdar, Barnali; Oswal, Rakesh G

    2015-01-01

    Background: Keratins are the most abundant proteins and are characteristic findings in many epithelial pathologies, making it a diagnostically important marker, both histopathologically and immunohistochemically. Since, immunohistochemistry is an expensive diagnostic tool, special stains to detect the degree of keratinization could serve as a faster and economic option. The aim of the present study was to compare the efficacy of special stains for keratin with standard hematoxylin and eosin stain (H and E). Objectives include: (i) To subject the diagnosed cases of keratin disorders to the selected special stains: Ayoub-shklar method, Dane-Herman method, Alcian blue –periodic acid Schiff ’s (PAS), rapid papanicolaou (PAP) and Gram’s stain. (ii) To compare the staining specificity and staining intensity of special stains with respect to routine hematoxylin and eosin (H and E) stain. (iii) To compare the efficacy of special stains to routine H and E stain in identification of the type of keratin present in the selected cases. Materials and Methods: A total of 80 cases of known pathology for keratin were retrieved from the department archive, which included 10 each of normal gingiva, hyperkeratosis, squamous papilloma, verrucous hyperplasia, verrucous carcinoma, well-differentiated squamous cell carcinoma, orthokeratinized odontogenic cyst and keratocystic odontogenic tumors. Six sections of 4 µ each from the paraffin blocks were made, stained with H and E and the special stains and these were evaluated by 2 pathologists based on the modified scoring criteria from Rahma Al-Maaini and Philip Bryant 2008. Results: The results were tabulated using Chi square and kappa statistics. The statistical values for identification of the type of keratinization was insignificant showing that ortho and parakeratinized epithelia could be correctly identified by both H and E as well as all the special stains. Furthermore, all the special stains showed a positive result and

  9. A standard tissue as a control for histochemical and immunohistochemical staining.

    PubMed

    Otali, D; Fredenburgh, J; Oelschlager, D K; Grizzle, W E

    2016-07-01

    The variable quality of histochemical and immunohistochemical staining of tissues may be attributed to pre-analytical and analytical variables. Both categories of variables frequently are undefined or inadequately controlled during specimen collection and preparation. Pre-analytical variables may alter the molecular composition of tissues, which results in variable staining; such variations may cause problems when different tissues are used as staining controls. We developed a standard tissue for use as a staining control. Our standard tissue contains five components: 1) nine combined human cell lines mixed with stroma from human spleen; 2) a squamous cancer cell line, A431; 3) fungus; 4) transverse sections of the mosquitofish and 5) normal human spleen. The first three components were embedded in HistoGel(™) and all components were processed to paraffin and used to construct a single standard paraffin block. The muscles of mosquitofish and arteries of the spleen are positive controls for eosin staining, while other tissues are useful for assessing hematoxylin staining. The mosquitofish tissues also are excellent controls for the Masson trichrome stain and all mucin-related histochemical stains that we tested. The goblet cells of the intestine and skin stained strongly with Alcian blue, pH 2.5 (AB-2.5), mucicarmine, colloidal iron, periodic acid Schiff (PAS) or PAS-hematoxylin (PASH) and combination stains such as colloidal iron-PASH. Cell lines were not useful for evaluating histochemical stains except for PASH. The splenic stroma was a useful control for AB-2.5; however, eosin and mucin stains stained cell lines poorly, probably due to their rapid growth and associated loss of some differentiated characteristics such as production of mucins. Nevertheless, the cell lines were a critical control for immunohistochemical stains. Immunostaining of specific cell lines was consistent with the presence of markers, e.g., EGFr in DU145 cells. The cell lines expressed a

  10. Interaction of water-soluble amino acid Schiff base complexes with bovine serum albumin: fluorescence and circular dichroism studies.

    PubMed

    Gharagozlou, Mehrnaz; Boghaei, Davar M

    2008-12-15

    Fluorescence spectroscopy in combination with circular dichroism (CD) spectroscopy were used to investigate the interaction of water-soluble amino acid Schiff base complexes, [Zn(L1,2)(phen)] where phen is 1,10-phenanthroline and H2L1,2 is amino acid Schiff base ligands, with bovine serum albumin (BSA) under the physiological conditions in phosphate buffer solution adjusted to pH 7.0. The quenching mechanism of fluorescence was suggested as static quenching according to the Stern-Volmer equation. Quenching constants were determined using the Stern-Volmer equation to provide a measure of the binding affinity between amino acid Schiff base complexes and BSA. The thermodynamic parameters DeltaG, DeltaH and DeltaS at different temperatures (298, 310 and 318K) were calculated. The results indicate that the hydrophobic and hydrogen bonding interactions play a major role in [Zn(L1)(phen)]-BSA association, whereas hydrophobic and electrostatic interactions participate a main role in [Zn(L2)(phen)]-BSA binding process. Binding studies concerning the number of binding sites and apparent binding constant Kb were performed by fluorescence quenching method. The distance R between the donor (BSA) and acceptor (amino acid Schiff base complexes) has been obtained utilizing fluorescence resonant energy transfer (FRET). Furthermore, CD spectra were used to investigate the structural changes of the BSA molecule with the addition of amino acid Schiff base complexes. The results indicate that the interaction of amino acid Schiff base complexes with BSA leads to changes in the secondary structure of the protein. Fractional contents of the secondary structure of BSA (f(alpha), f(beta), f(turn) and f(random)) were calculated with and without amino acid Schiff base complexes utilizing circular dichroism spectroscopy. Our results clarified that amino acid Schiff base complexes could bind to BSA and be effectively transported and eliminated in the body, which could be a useful guideline for

  11. Synthesis and antimicrobial activities of new higher amino acid Schiff base derivatives of 6-aminopenicillanic acid and 7-aminocephalosporanic acid

    NASA Astrophysics Data System (ADS)

    Özdemir (nee Güngör), Özlem; Gürkan, Perihan; Özçelik, Berrin; Oyardı, Özlem

    2016-02-01

    Novel β-lactam derivatives (1c-3c) (1d-3d) were produced by using 6-aminopenicillanic acid (6-APA), 7-aminocephalosporanic acid (7-ACA) and the higher amino acid Schiff bases. The synthesized compounds were characterized by elemental analysis, IR, 1H/13C NMR and UV-vis spectra. Antibacterial activities of all the higher amino acid Schiff bases (1a-3a) (1b-3b) and β-lactam derivatives were screened against three gram negative bacteria (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Acinetobacter baumannii RSKK 02026), three gram positive bacteria (Staphylococcus aureus ATCC 25923, Enterococcus faecalis ATCC 07005, Bacillus subtilis ATCC 6633) and their drug-resistant isolates by using broth microdilution method. Two fungi (Candida albicans and Candida krusei) were used for antifungal activity.

  12. Gram stain

    MedlinePlus

    A Gram stain is a test used to identify bacteria. It is one of the most common ways to ... Urethral discharge Gram stain; Feces Gram stain; Stool Gram stain; Joint fluid Gram stain; Pericardial fluid Gram stain; Gram stain of ...

  13. DNA interaction with octahedral and square planar Ni(II) complexes of aspartic-acid Schiff-bases

    NASA Astrophysics Data System (ADS)

    Sallam, S. A.; Orabi, A. S.; Abbas, A. M.

    2011-12-01

    Ni(II) complexes of (S,E)-2-(2-OHbenzilydene)aspartic acid; (S,E)-2-(2,3-diOHbenzilydene)aspartic acid-; (S,E)-2-(2,4-diOH-benzilydene)aspartic acid; (S,E)-2-(2,5-diOHbenzilydene)aspartic acid and (S,E)-2-((2-OHnaphthalene-1-yl)methylene)aspartic acid Schiff-bases have been synthesized by template method in ethanol or ammonia media. They were characterized by elemental analyses, conductivity measurements, magnetic moment, UV, IR and 1H nmr spectra as well as thermal analysis (TG, DTG, DTA). The Schiff-bases are dibasic tridentate or tetradentate donors and the complexes have square planar and octahedral structures. The complexes decompose in two or three steps where kinetic and thermodynamic parameters of the decomposition steps were computed. The interactions of the formed complexes with FM-DNA were monitored by UV and fluorescence spectroscopy.

  14. De-staining and re-staining mucins in formalin fixed paraffin sections.

    PubMed

    Smith, A A; Glickfield, I

    2011-04-01

    Re-staining of formalin fixed paraffin sections sometimes is required and this requires prior de-staining. Some simple and effective protocols for de-staining are described. Mucihematoxylin and mucicarmine can be removed with acid alcohol. Zirconyl hematoxylin can be removed with periodic acid or Sinha's fixative. Alcian blue can be removed with 5% trifluoroacetic acid in dichloromethane. Colloidal iron can be bleached in 1% household bleach in alcohol. PAS can be removed with hydrogen peroxide or ammonium hydroxide. With few exceptions, de-stained sections can be re-stained with mucihematoxylin, PAS or Gabe's trichrome. PMID:20001228

  15. Wood stains

    MedlinePlus

    The harmful substances in wood stains are hydrocarbons, or substances that contain only carbon and hydrogen. Other harmful ingredients may include: Alcohol Alkanes Cyclo alkanes Glycol ether Corrosives, such as sodium ...

  16. Gram Stain

    MedlinePlus

    ... definitively identify the cause of infection. Fungi , including yeast, may also be detected with a Gram stain. ^ ... white blood cells Fungi (in the form of yeasts or molds) may be seen on a Gram ...

  17. 2-Phenylacetylenesulfonamide (PAS) induces p53-independent apoptotic killing of B-chronic lymphocytic leukemia (CLL) cells.

    PubMed

    Steele, Andrew J; Prentice, Archibald G; Hoffbrand, A Victor; Yogashangary, Birunthini C; Hart, Stephen M; Lowdell, Mark W; Samuel, Edward R; North, Janet M; Nacheva, Elisabeth P; Chanalaris, Anastasios; Kottaridis, Panagiotis; Cwynarski, Kate; Wickremasinghe, R Gitendra

    2009-08-01

    We studied the actions of 2-phenylacetylenesulfonamide (PAS) on B-chronic lymphocytic leukemia (CLL) cells. PAS (5-20 microM) initiated apoptosis within 24 hours, with maximal death at 48 hours asassessed by morphology, cleavage of poly(ADP-ribose) polymerase (PARP), caspase 3 activation, and annexin V staining. PAS treatment induced Bax proapoptotic conformational change, Bax movement from the cytosol to the mitochondria, and cytochrome c release, indicating that PAS induced apoptosis via the mitochondrial pathway. PAS induced approximately 3-fold up-regulation of proapoptotic Noxa protein and mRNA levels. In addition, Noxa was found unexpectedly to be bound to Bcl-2 in PAS-treated cells. PAS treatment of CLL cells failed to up-regulate p53, suggesting that PAS induced apoptosis independently of p53. Furthermore, PAS induced apoptosis in CLL isolates with p53 gene deletion in more than 97% of cells. Normal B lymphocytes were as sensitive to PAS-induced Noxa up-regulation and apoptosis as were CLL cells. However, both T lymphocytes and bone marrow hematopoietic progenitor cells were relatively resistant to PAS. Our data suggest that PAS may represent a novel class of drug that induces apoptosis in CLL cells independently of p53 status by a mechanism involving Noxa up-regulation. PMID:19515722

  18. Ford Partnership for Advanced Studies (Ford PAS)

    ERIC Educational Resources Information Center

    Carrier, Cheryl

    2006-01-01

    This article features the Ford Partnership for Advanced Studies (Ford PAS) program that helps students gain knowledge about real-life business issues. Ford PAS is an educational program that combines college-prep academics with the critical 21st century skills students will need to succeed in college and in the workplace. The Ford PAS program uses…

  19. A procedure for Alcian blue staining of mucins on polyvinylidene difluoride membranes.

    PubMed

    Dong, Weijie; Matsuno, Yu-ki; Kameyama, Akihiko

    2012-10-16

    The isolation and characterization of mucins are critically important for obtaining insight into the molecular pathology of various diseases, including cancers and cystic fibrosis. Recently, we developed a novel membrane electrophoretic method, supported molecular matrix electrophoresis (SMME), which separates mucins on a polyvinylidene difluoride (PVDF) membrane impregnated with a hydrophilic polymer. Alcian blue staining is widely used to visualize mucopolysaccharides and acidic mucins on both blotted membranes and SMME membranes; however, this method cannot be used to stain mucins with a low acidic glycan content. Meanwhile, periodic acid-Schiff staining can selectively visualize glycoproteins, including mucins, but is incompatible with glycan analysis, which is indispensable for mucin characterizations. Here we describe a novel staining method, designated succinylation-Alcian blue staining, for visualizing mucins on a PVDF membrane. This method can visualize mucins regardless of the acidic residue content and shows a sensitivity 2-fold higher than that of Pro-Q Emerald 488, a fluorescent periodate Schiff-base stain. Furthermore, we demonstrate the compatibility of this novel staining procedure with glycan analysis using porcine gastric mucin as a model mucin. PMID:22950532

  20. Water soluble and efficient amino acid Schiff base receptor for reversible fluorescence turn-on detection of Zn2+ ions: Quantum chemical calculations and detection of bacteria

    NASA Astrophysics Data System (ADS)

    Subha, L.; Balakrishnan, C.; Natarajan, Satheesh; Theetharappan, M.; Subramanian, Balanehru; Neelakantan, M. A.

    2016-01-01

    An amino acid Schiff base (R) capable of recognizing Zn2+ ions selectively and sensitively in an aqueous medium was prepared and characterized. Upon addition of Zn2+ ions, the receptor exhibits fluorescence intensity enhancements (~ 40 fold) at 460 nm (quantum yield, Φ = 0.05 for R and Φ = 0.18 for R-Zn2+) and can be detected by naked eye under UV light. The receptor can recognize the Zn2+ (1.04 × 10- 8 M) selectively for other metal ions in the pH range of 7.5-11. The Zn2+ chelation with R decreases the loss of energy through non-radiative transition and leads to fluorescence enhancement. The binding mode of the receptor with Zn2+ was investigated by 1H NMR titration and further validated by ESI-MS. The elemental color mapping and SEM/EDS analysis were also used to study the binding of R with Zn2+. Density functional theory calculations were carried out to understand the binding mechanism. The receptor was applied as a microbial sensor for Escherichia coli and Staphylococcus aureus.

  1. Regulation and function of yeast PAS kinase

    PubMed Central

    Grose, Julianne H.; Sundwall, Eleanor; Rutter, Jared

    2016-01-01

    The inability to coordinate cellular metabolic processes with the cellular and organismal nutrient environment leads to a variety of disorders, including diabetes and obesity. Nutrient-sensing protein kinases, such as AMPK and mTOR, play a pivotal role in metabolic regulation and are promising therapeutic targets for the treatment of disease. In this Extra View, we describe another member of the nutrient-sensing protein kinase group, PAS kinase, which plays a role in the regulation of glucose utilization in both mammals and yeast. PAS kinase deficient mice are resistant to high fat diet-induced weight gain, insulin resistance and hepatic triglyceride hyperaccumulation, suggesting a role for PAS kinase in the regulation of glucose and lipid metabolism in mammals. Likewise, PAS kinase deficient yeast display altered glucose partitioning, favoring glycogen biosynthesis at the expense of cell wall biosynthesis. As a result, PAS kinase deficient yeast are sensitive to cell wall perturbing agents. This partitioning of glucose in response to PAS kinase activation is due to phosphorylation of Ugp1, the enzyme primarily responsible for UDP-glucose production. The two yeast PAS kinase homologs, Psk1 and Psk2, are activated by two stimuli, cell integrity stress and nonfermentative carbon sources. We review what is known about yeast PAS kinase and describe a genetic screen that may help elucidate pathways involved in PAS kinase activation and function. PMID:19440050

  2. Port-wine stain

    MedlinePlus

    ... wine stains, including freezing, surgery, radiation, and tattooing. Laser therapy is most successful in eliminating port-wine stains. ... Prognosis) Stains on the face respond better to laser therapy than those on the arms, legs, or middle ...

  3. Joint fluid Gram stain

    MedlinePlus

    Gram stain of joint fluid ... result means no bacteria are present on the Gram stain. Normal value ranges may vary slightly among ... Abnormal results mean bacteria were seen on the Gram stain. This may be a sign of a ...

  4. Port-Wine Stain

    MedlinePlus

    ... and rashes clinical tools newsletter | contact Share | Port-Wine Stain A parent's guide for infants and babies ... a three-month-old infant with a port-wine stain. Overview A port-wine stain is a ...

  5. Cellular and computational studies of proteasome inhibition and apoptosis induction in human cancer cells by amino acid Schiff base–copper complexes

    PubMed Central

    Zuo, Jian; Bi, Caifeng; Fan, Yuhua; Buac, Daniela; Nardon, Chiara; Daniel, Kenyon G.; Dou, Q. Ping

    2013-01-01

    Proliferation and apoptosis pathways are tightly regulated in a cell by the ubiquitin–proteasome system (UPS) and alterations in the UPS may result in cellular transformation or other pathological conditions. Indeed, the proteasome is often found to be overactive in cancer cells. It has also been found that cancer cells are more sensitive to proteasome inhibition than normal cells, and therefore proteasome inhibitors are pursued as antitumor drugs. The use of the proteasome inhibitor Bortezomib for treatment of multiple myeloma and mantle cell lymphoma has proved this principle. Recent studies have suggested that copper complexes can inhibit proteasome activity and induce apoptosis in some human cancer cells. However, the involved molecular mechanism is unknown. In this study, we investigated the biological activities of four amino acid Schiff base–copper(II) complexes by using human breast (MDA-MB-231 and MCF-7) and prostate (PC-3) cancer cells. The complexes C1 and C3, but not their counterparts C2 and C4, inhibit the chymotrypsin-like activity of purified 20S proteasome and human cancer cellular 26S proteasome, cause accumulation of proteasome target proteins Bax and IκB-α, and induce growth inhibition and apoptosis in concentration- and time-dependent manners. Docking analysis shows that C1, but not C2 has hydrophobic, pi–pi, pi–cation and hydrogen bond interactions with the proteasomal chymotrypsin-like pocket and could stably fit into the S3 region, leading to specific inhibition. Our study has identified the mechanism of action of these copper complexes on inhibiting tumor cell proteasome and suggested their great potential as novel anticancer agents. PMID:23142973

  6. Acid-fast stain

    MedlinePlus

    The acid-fast stain is a laboratory test that determines if a sample of tissue, blood, or other body ... dye. The slide is then washed with an acid solution and a different stain is applied. Bacteria ...

  7. Acid-fast stain

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003766.htm Acid-fast stain To use the sharing features on this page, please enable JavaScript. The acid-fast stain is a laboratory test that determines ...

  8. Port-wine stain

    MedlinePlus

    A port-wine stain is a birthmark in which swollen blood vessels create a reddish-purplish discoloration of the skin. ... Port-wine stains occur in about 3 out of 1,000 people. In rare cases, port-wine stains are ...

  9. Black stain - a review.

    PubMed

    Ronay, Valerie; Attin, Thomas

    2011-01-01

    The purpose of this review was to summarise the fundamentals about black stain, its diagnosis and possible differential diagnoses as well as its microbiology and therapy. In addition, various studies investigating the relationship between black stain and dental caries are examined. Many studies report lower caries prevalence in children with black stain, but this finding could not be confirmed by all authors. Also, a negative relation between degree of staining and caries severity has been described. Reasons for these results are not yet clear but it was speculated that they are related to the specific oral microflora described in black stain-affected individuals. PMID:21594205

  10. Gram stain of urethral discharge

    MedlinePlus

    Urethral discharge Gram stain ... microscope slide. A series of stains called a Gram stain is applied to the specimen. The stained ... culture ) should be performed in addition to the gram stain. More sophisticated diagnostic tests (such as PCR ...

  11. Dramatic Stained Glass.

    ERIC Educational Resources Information Center

    Prater, Michael

    2002-01-01

    Describes an art project that is appropriate for students in fifth through twelfth grade in which they create Gothic-style stained-glass windows. Discusses how college students majoring in elementary education created stained-glass windows. Addresses how to adapt this lesson for younger students. (CMK)

  12. Setting the PAS, the role of circadian PAS domain proteins during environmental adaptation in plants

    PubMed Central

    Vogt, Julia H. M.; Schippers, Jos H. M.

    2015-01-01

    The per-ARNT-sim (PAS) domain represents an ancient protein module that can be found across all kingdoms of life. The domain functions as a sensing unit for a diverse array of signals, including molecular oxygen, small metabolites, and light. In plants, several PAS domain-containing proteins form an integral part of the circadian clock and regulate responses to environmental change. Moreover, these proteins function in pathways that control development and plant stress adaptation responses. Here, we discuss the role of PAS domain-containing proteins in anticipation, and adaptation to environmental changes in plants. PMID:26217364

  13. The J-PAS filter system

    NASA Astrophysics Data System (ADS)

    Marin-Franch, Antonio; Taylor, Keith; Cenarro, Javier; Cristobal-Hornillos, David; Moles, Mariano

    2015-08-01

    J-PAS (Javalambre-PAU Astrophysical Survey) is a Spanish-Brazilian collaboration to conduct a narrow-band photometric survey of 8500 square degrees of northern sky using an innovative filter system of 59 filters, 56 relatively narrow-band (FWHM=14.5 nm) filters continuously populating the spectrum between 350 to 1000nm in 10nm steps, plus 3 broad-band filters. This filter system will be able to produce photometric redshifts with a precision of 0.003(1 + z) for Luminous Red Galaxies, allowing J-PAS to measure the radial scale of the Baryonic Acoustic Oscillations. The J-PAS survey will be carried out using JPCam, a 14-CCD mosaic camera using the new e2v 9k-by-9k, 10μm pixel, CCDs mounted on the JST/T250, a dedicated 2.55m wide-field telescope at the Observatorio Astrofísico de Javalambre (OAJ) near Teruel, Spain. The filters will operate in a fast (f/3.6) converging beam. The requirements for average transmissions greater than 85% in the passband, <10-5 blocking from 250 to 1050nm, steep bandpass edges and high image quality impose significant challenges for the production of the J-PAS filters that have demanded the development of new design solutions. This talk presents the J-PAS filter system and describes the most challenging requirements and adopted design strategies. Measurements and tests of the first manufactured filters are also presented.

  14. Pleural fluid Gram stain

    MedlinePlus

    Gram stain of pleural fluid ... lungs fill a person's chest with air. If fluid builds up in the space outside the lungs ... chest, it can cause many problems. Removing the fluid can relieve a person's breathing problems and help ...

  15. Les etoiles qui ne veulent pas vieillir

    NASA Astrophysics Data System (ADS)

    Bonnet-Bidaud, J. M.

    1995-12-01

    Qu'est-ce qui fait courir Jean-Claude Pecker ? Ses travaux sur les atmospheres stellaires qu'il poursuit aujourd'hui ? Son combat pour les droits de l'homme, ou contre le sceau du secret qui pese encore sur la recherche fondamentale ? Tout a la fois. Pour cette figure emblematique de l'astrophysique francaise, aujourd'hui a la retraite, pas question de raccrocher les armes...

  16. Apparatus Would Stain Microscope Slides

    NASA Technical Reports Server (NTRS)

    Breeding, James D.

    1993-01-01

    Proposed apparatus meters specific amounts of fluid out of containers at specific times to stain microscope slides. Intended specifically for semiautomated staining of microbiological and hematological samples in microgravity, leakproof apparatus used in other environments in which technicians have little time to allocate to staining procedures and/or exposure to toxic staining agents or to micro-organisms to be stained hazardous. Apparatus adapted to perform almost any staining procedure and accommodates multiple staining reagents, useful for small or remote clinical laboratories.

  17. Stained-Glass Pastels

    ERIC Educational Resources Information Center

    Laird, Shirley

    2009-01-01

    The author has always liked the look of stained-glass windows. Usually the designs are simplified and the shapes are easier for younger students to draw. This technique seemed to be the perfect place for her fifth-graders to try their hand at color mixing. The smaller spaces and simple shapes were just what she needed for this group. Her students…

  18. "Stained Glass" Landscape Windows

    ERIC Educational Resources Information Center

    Vannata, Janine

    2008-01-01

    Both adults and children alike marvel at the grand vivid stained-glass windows created by American artist Louis Comfort Tiffany. Today he is commonly recognized as one of America's most influential designers and artists throughout the last nineteenth and early twentieth century. In the lesson described in this article, students created their own…

  19. Port-Wine Stains

    MedlinePlus

    ... upsetting for kids, especially if they're large, dark, or on the face. And any birthmark can take a toll on a child's self-confidence, no matter how large or small the mark might be. The good news is that lasers (highly concentrated light energy) can make many kids' port-wine stains much ...

  20. Shimmering Stained Glass.

    ERIC Educational Resources Information Center

    Simon, Gail Murray

    1998-01-01

    Presents an art lesson for fifth- and sixth-graders where they create a translucent design of colored cellophane on black paper inspired by the stained-glass windows of the Middle Ages and the artwork of Lewis Comfort Tiffany. Enables the students to become crafts people rather than just observers of the past. (CMK)

  1. Fluorescein eye stain

    MedlinePlus

    ... eye. The health care provider then shines a blue light at your eye. Any problems on the surface of the cornea will be stained by the dye and appear green under the blue light. The provider can determine the location and ...

  2. Joint fluid Gram stain

    MedlinePlus

    Gram stain of joint fluid ... A sample of joint fluid is needed. The fluid sample is sent to a lab where a small drop is placed in a ... on how to prepare for the removal of joint fluid, see joint fluid aspiration .

  3. Water soluble and efficient amino acid Schiff base receptor for reversible fluorescence turn-on detection of Zn²⁺ ions: Quantum chemical calculations and detection of bacteria.

    PubMed

    Subha, L; Balakrishnan, C; Natarajan, Satheesh; Theetharappan, M; Subramanian, Balanehru; Neelakantan, M A

    2016-01-15

    An amino acid Schiff base (R) capable of recognizing Zn(2+) ions selectively and sensitively in an aqueous medium was prepared and characterized. Upon addition of Zn(2+) ions, the receptor exhibits fluorescence intensity enhancements (~40 fold) at 460 nm (quantum yield, Φ=0.05 for R and Φ=0.18 for R-Zn(2+)) and can be detected by naked eye under UV light. The receptor can recognize the Zn(2+) (1.04×10(-8) M) selectively for other metal ions in the pH range of 7.5-11. The Zn(2+) chelation with R decreases the loss of energy through non-radiative transition and leads to fluorescence enhancement. The binding mode of the receptor with Zn(2+) was investigated by (1)H NMR titration and further validated by ESI-MS. The elemental color mapping and SEM/EDS analysis were also used to study the binding of R with Zn(2+). Density functional theory calculations were carried out to understand the binding mechanism. The receptor was applied as a microbial sensor for Escherichia coli and Staphylococcus aureus. PMID:26318699

  4. Length of stain dosimeter

    NASA Technical Reports Server (NTRS)

    Lueck, Dale E. (Inventor)

    1994-01-01

    Payload customers for the Space Shuttle have recently expressed concerns about the possibility of their payloads at an adjacent pad being contaminated by plume effluents from a shuttle at an active pad as they await launch on an inactive pad. As part of a study to satisfy such concerns a ring of inexpensive dosimeters was deployed around the active pad at the inter-pad distance. However, following a launch, dosimeters cannot be read for several hours after the exposure. As a consequence factors such as different substrates, solvent systems, and possible volatilization of HCl from the badges were studied. This observation led to the length of stain (LOS) dosimeters of this invention. Commercial passive LOS dosimeters are sensitive only to the extent of being capable of sensing 2 ppm to 20 ppm if the exposure is 8 hours. To map and quantitate the HCl generated by Shuttle launches, and in the atmosphere within a radius of 1.5 miles from the active pad, a sensitivity of 2 ppm HCl in the atmospheric gases on an exposure of 5 minutes is required. A passive length of stain dosimeter has been developed having a sensitivity rendering it capable of detecting a gas in a concentration as low as 2 ppm on an exposure of five minutes.

  5. Blood stain pattern analysis.

    PubMed

    Peschel, O; Kunz, S N; Rothschild, M A; Mützel, E

    2011-09-01

    Bloodstain pattern analysis (BPA) refers to the collection, categorization and interpretation of the shape and distribution of bloodstains connected with a crime. These kinds of stains occur in a considerable proportion of homicide cases. They offer extensive information and are an important part of a functional, medically and scientifically based reconstruction of a crime. The following groups of patterns can essentially be distinguished: dripped and splashed blood, projected blood, impact patterns, cast-off stains, expirated and transferred bloodstains. A highly qualified analysis can help to estimate facts concerning the location, quality and intensity of an external force. A sequence of events may be recognized, and detailed questions connected with the reconstruction of the crime might be answered. In some cases, BPA helps to distinguish between accident, homicide and suicide or to identify bloodstains originating from a perpetrator. BPA is based on systematic training, a visit to the crime scene or alternatively good photographic documentation, and an understanding and knowledge of autopsy findings or statements made by the perpetrator and/or victim. A BPA working group has been established within the German Society of Legal Medicine aiming to put the knowledge and practical applications of this subdiscipline of forensic science on a wider basis. PMID:21069481

  6. Gram stain of tissue biopsy

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003453.htm Gram stain of tissue biopsy To use the sharing features on this page, please enable JavaScript. Gram stain of tissue biopsy test involves using crystal ...

  7. Feasibility of classification of clay minerals by using PAS

    NASA Astrophysics Data System (ADS)

    Honda, Y.; Yoshida, Y.; Akiyama, Y.; Nishijima, S.

    2015-06-01

    After the nuclear power plant disaster, the evaluation of radioactive Cs kept in soil, especially in clay minerals and the elucidation of its movement are urgent subjects to promote decontamination. It is known that the extractable level of Cs depends on the sort of clay minerals. We tried to find the characteristics of clay minerals belonging to phillosilicate group using positron annihilation spectroscopy (PAS) and the relationship between the results of PAS and the amounts of substantially extracted Cs from the clay minerals. The results showed that each clay mineral was found to be distinguishable from other clay minerals by PAS and the extraction rate of Cs was different among those clay minerals, however the direct correlation between the results of PAS and the extraction rates of Cs was not found.

  8. Competitive concerns bring PAs, NPs into triage area.

    PubMed

    2008-06-01

    The ED at Harborview Medical Center in Seattle has proven new competition doesn't have to mean a loss of business or staff members. They moved physician assistants (PAs) and nurse practitioners (NPs) into triage to compete with urgent care clinics. When you have a hard time attracting midlevel practitioners, examine the local market to see who is recruiting them. Create a small, separate area in triage where midlevel practitioners can treat patients with less serious conditions. Be flexible with the number of hours the PAs and NPs work. Be prepared to expand their hours as demand increases. PMID:18807298

  9. Array tomography: imaging stained arrays.

    PubMed

    Micheva, Kristina D; O'Rourke, Nancy; Busse, Brad; Smith, Stephen J

    2010-11-01

    Array tomography is a volumetric microscopy method based on physical serial sectioning. Ultrathin sections of a plastic-embedded tissue are cut using an ultramicrotome, bonded in an ordered array to a glass coverslip, stained as desired, and imaged. The resulting two-dimensional image tiles can then be reconstructed computationally into three-dimensional volume images for visualization and quantitative analysis. The minimal thickness of individual sections permits high-quality rapid staining and imaging, whereas the array format allows reliable and convenient section handling, staining, and automated imaging. Also, the physical stability of the arrays permits images to be acquired and registered from repeated cycles of staining, imaging, and stain elution, as well as from imaging using multiple modalities (e.g., fluorescence and electron microscopy). Array tomography makes it possible to visualize and quantify previously inaccessible features of tissue structure and molecular architecture. However, careful preparation of the tissue is essential for successful array tomography; these steps can be time-consuming and require some practice to perfect. In this protocol, tissue arrays are imaged using conventional wide-field fluorescence microscopy. Images can be captured manually or, with the appropriate software and hardware, the process can be automated. PMID:21041399

  10. Automated single-slide staining system

    NASA Technical Reports Server (NTRS)

    Mills, S. M.; Wilkins, J. R.

    1974-01-01

    Apparatus developed to Gram-stain single slides automatically is flexible enough to accommodate other types of staining procedures. Method frees operator and eliminates necessity for subjective evaluations as to length of staining or decolorizing time.

  11. Whole Blood Cell Staining Device

    NASA Technical Reports Server (NTRS)

    Sams, Clarence F.; Clift, Vaughan L.; McDonald, Kelly E.

    2000-01-01

    An apparatus and method for staining particular cell markers is disclosed. The apparatus includes a flexible tube that is reversibly pinched into compartments with one or more clamps. Each compartment of the tube contains a separate reagent and is in selective fluid communication with adjoining compartments.

  12. Procedures and acceptance criteria for PAS-1 cask inspections

    SciTech Connect

    Mercado, J.E.

    1998-09-09

    The procedures and acceptance criteria that comprise this document were prepared to support a one-time test to certify two PAS-1 casks in accordance with US Department of Energy Certificate of Compliance US A/9184/B(U), which was issued in 1998. The specific inspections addressed in this document are the visual weld inspection and a dimensional inspection of the primary containment vessel.

  13. Methods for chromosome-specific staining

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel

    1995-01-01

    Methods and compositions for chromosome-specific staining are provided. Compositions comprise heterogenous mixtures of labeled nucleic acid fragments having substantially complementary base sequences to unique sequence regions of the chromosomal DNA for which their associated staining reagent is specific. Methods include methods for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes.

  14. Salt stains from evaporating droplets.

    PubMed

    Shahidzadeh, Noushine; Schut, Marthe F L; Desarnaud, Julie; Prat, Marc; Bonn, Daniel

    2015-01-01

    The study of the behavior of sessile droplets on solid substrates is not only associated with common everyday phenomena, such as the coffee stain effect, limescale deposits on our bathroom walls , but also very important in many applications such as purification of pharmaceuticals, de-icing of airplanes, inkjet printing and coating applications. In many of these processes, a phase change happens within the drop because of solvent evaporation, temperature changes or chemical reactions, which consequently lead to liquid to solid transitions in the droplets. Here we show that crystallization patterns of evaporating of water drops containing dissolved salts are different from the stains reported for evaporating colloidal suspensions. This happens because during the solvent evaporation, the salts crystallize and grow during the drying. Our results show that the patterns of the resulting salt crystal stains are mainly governed by wetting properties of the emerging crystal as well as the pathway of nucleation and growth, and are independent of the evaporation rate and thermal conductivity of the substrates. PMID:26012481

  15. Salt stains from evaporating droplets

    PubMed Central

    Shahidzadeh, Noushine; Schut, Marthe F. L.; Desarnaud, Julie; Prat, Marc; Bonn, Daniel

    2015-01-01

    The study of the behavior of sessile droplets on solid substrates is not only associated with common everyday phenomena, such as the coffee stain effect, limescale deposits on our bathroom walls , but also very important in many applications such as purification of pharmaceuticals, de-icing of airplanes, inkjet printing and coating applications. In many of these processes, a phase change happens within the drop because of solvent evaporation, temperature changes or chemical reactions, which consequently lead to liquid to solid transitions in the droplets. Here we show that crystallization patterns of evaporating of water drops containing dissolved salts are different from the stains reported for evaporating colloidal suspensions. This happens because during the solvent evaporation, the salts crystallize and grow during the drying. Our results show that the patterns of the resulting salt crystal stains are mainly governed by wetting properties of the emerging crystal as well as the pathway of nucleation and growth, and are independent of the evaporation rate and thermal conductivity of the substrates. PMID:26012481

  16. Geodynamic Research at the Department of Planetary Geodesy, SRC PAS

    NASA Astrophysics Data System (ADS)

    Brzeziński, Aleksander; Jóźwik, Mieczysław; Kaczorowski, Marek; Kalarus, Maciej; Kasza, Damian; Kosek, Wiesław; Nastula, Jolanta; Szczerbowski, Zbigniew; Wińska, Małgorzata; Wronowski, Roman; Zdunek, Ryszard; Zieliński, Janusz B.

    2016-06-01

    The Department of Planetary Geodesy of the Space Research Centre PAS has been conducting research on a broad spectrum of problems within a field of global dynamics of the Earth. In this report we describe the investigations on selected subjects concerning polar motion (modeling and geophysical interpretation of the Chandler wobble, hydrological excitation of seasonal signals, search for optimal prediction methods), tectonic activity in the region of the Książ Geodynamic Laboratory of the SRC, and finally the new joint Polish-Italian project GalAc analyzing feasibility and usefulness of equipping second-generation Galileo satellites with accelerometers.

  17. Delineating PAS-HAMP interaction surfaces and signalling-associated changes in the aerotaxis receptor Aer.

    PubMed

    Garcia, Darysbel; Watts, Kylie J; Johnson, Mark S; Taylor, Barry L

    2016-04-01

    The Escherichia coli aerotaxis receptor, Aer, monitors cellular oxygen and redox potential via FAD bound to a cytosolic PAS domain. Here, we show that Aer-PAS controls aerotaxis through direct, lateral interactions with a HAMP domain. This contrasts with most chemoreceptors where signals propagate along the protein backbone from an N-terminal sensor to HAMP. We mapped the interaction surfaces of the Aer PAS, HAMP and proximal signalling domains in the kinase-off state by probing the solvent accessibility of 129 cysteine substitutions. Inaccessible PAS-HAMP surfaces overlapped with a cluster of PAS kinase-on lesions and with cysteine substitutions that crosslinked the PAS β-scaffold to the HAMP AS-2 helix. A refined Aer PAS-HAMP interaction model is presented. Compared to the kinase-off state, the kinase-on state increased the accessibility of HAMP residues (apparently relaxing PAS-HAMP interactions), but decreased the accessibility of proximal signalling domain residues. These data are consistent with an alternating static-dynamic model in which oxidized Aer-PAS interacts directly with HAMP AS-2, enforcing a static HAMP domain that in turn promotes a dynamic proximal signalling domain, resulting in a kinase-off output. When PAS-FAD is reduced, PAS interaction with HAMP is relaxed and a dynamic HAMP and static proximal signalling domain convey a kinase-on output. PMID:26713609

  18. Methods for chromosome-specific staining

    DOEpatents

    Gray, J.W.; Pinkel, D.

    1995-09-05

    Methods and compositions for chromosome-specific staining are provided. Compositions comprise heterogeneous mixtures of labeled nucleic acid fragments having substantially complementary base sequences to unique sequence regions of the chromosomal DNA for which their associated staining reagent is specific. Methods include ways for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes. 3 figs.

  19. Golgi-Cox Staining Step by Step

    PubMed Central

    Zaqout, Sami; Kaindl, Angela M.

    2016-01-01

    Golgi staining remains a key method to study neuronal morphology in vivo. Since most protocols delineating modifications of the original staining method lack details on critical steps, establishing this method in a laboratory can be time-consuming and frustrating. Here, we describe the Golgi-Cox staining in such detail that should turn the staining into an easily feasible method for all scientists working in the neuroscience field. PMID:27065817

  20. Comparison of two different passive air samplers (PUF-PAS versus SIP-PAS) to determine time-integrated average air concentration of volatile hydrophobic organic pollutants

    NASA Astrophysics Data System (ADS)

    Kim, Seung-Kyu; Park, Jong-Eun

    2014-06-01

    Despite remarkable achievements with r some chemicals, a field-measurement technique has not been advanced for volatile hydrophobic organic chemicals (HOCs) that are the subjects of international concern. This study assesses the applicability of passive air sampling (PAS) by comparing PUF-PAS and its modified SIP-PAS which was made by impregnating XAD-4 powder into PUF, overviewing the principles of PAS, screening sensitive parameters, and determining the uncertainty range of PAS-derived concentration. The PAS air sampling rate determined in this study, corrected by a co-deployed low-volume active air sampler (LAS) for neutral PFCs as model chemicals, was ˜1.2 m3 day-1. Our assessment shows that the improved sorption capacity in a SIP lengthens PAS deployment duration by expanding the linear uptake range and then enlarges the effective air sampling volume and detection frequency of chemicals at trace level. Consequently, volatile chemicals can be collected during sufficiently long times without reaching equilibrium when using SIP, while this is not possible for PUF. The most sensitive parameter to influence PAS-derived CA was an air-side mass transfer coefficient (kA), implying the necessity of spiking depuration chemicals (DCs) because this parameter is strongly related with meteorological conditions. Uncertainty in partition coefficients (KPSM-A or KOA) influences PAS-derived CA to a greater extent with regard to lower KPSM-A chemicals. Also, the PAS-derived CA has an uncertainty range of a half level to a 3-fold higher level of the calculated one. This work is expected to establish solid grounds for the improvement of field measurement technique of HOCs.

  1. Antibody Staining in Drosophila Germaria.

    PubMed

    Lie-Jensen, Anette; Haglund, Kaisa

    2016-01-01

    Drosophila oogenesis is a powerful model for studying a wide spectrum of cellular and developmental processes in vivo. Oogenesis starts in a specialized structure called the germarium, which harbors the stem cells for both germ and somatic cells. The germarium produces egg chambers, each of which will develop into an egg. Active areas of research in Drosophila germaria include stem cell self-renewal, division, and maintenance, cell cycle control and differentiation, oocyte specification, intercellular communication, and signaling, among others. The solid knowledge base, the genetic tractability of the Drosophila model, as well as the availability and fast development of tools and imaging techniques for oogenesis research ensure that studies in this model will keep being instrumental for novel discoveries within cell and developmental biology also in the future. This chapter focuses on antibody staining in Drosophila germaria and provides a protocol for immunostaining as well as an overview of commonly used antibodies for visualization of different cell types and cellular structures. The protocol is well-suited for subsequent confocal microscopy analyses, and in addition we present key adaptations of the protocol that are useful when performing structured illumination microscopy (SIM) super-resolution imaging. PMID:27557571

  2. Quantitative studies of immunofluorescent staining

    PubMed Central

    Wick, G.; Beutner, E. H.

    1970-01-01

    The antiperinuclear factor (APF) is found in a high percentage of sera from patients with rheumatoid arthritis. It can be demonstrated by direct immunofluorescence using the keratohyaline granules of human buccal mucosa as antigenic substrate. Mixing of some normal goat sera with an APF positive serum from a patient with rheumatoid arthritis resulted in an inhibition of the APF titre of the patient's serum. However, there was no clear cut correlation between the APF-positivity of normal goat sera and their inhibitory effect on the APF-reactivity of a human rheumatoid arthritis patient's serum. In reciprocal screening tests the human rheumatoid arthritis serum blocked only one of the APF-reactive goat sera. The reciprocal blocking activity of this goat serum and the patient's serum could be more exactly evaluated by the use of chessboard titrations in an indirect immunofluorescence blocking test. This test consisted of mixing equal volumes of serial dilutions of a goat serum and the patient's serum and subsequent examination of the mixtures for APF using an anti-human IgG conjugate and an anti-goat immunoglobulin conjugate, respectively. The results point to an antibody nature for the APF in preimmune, normal goat sera and to the value of chessboard titrations of this type in demonstrating the identity, non-identity, partial identity (or very close proximity of antigenic determinants) of the antibodies in different antisera which cannot be distinguished by their immunofluorescent staining patterns. ImagesFIG. 1FIG. 2 PMID:4913803

  3. Multi-PAS domain-mediated protein oligomerization of PpsR from Rhodobacter sphaeroides

    SciTech Connect

    Heintz, Udo; Meinhart, Anton; Winkler, Andreas

    2014-03-01

    Crystal structures of two truncated variants of the transcription factor PpsR from R. sphaeroides are presented that enabled the phasing of a triple PAS domain construct. Together, these structures reveal the importance of α-helical PAS extensions for multi-PAS domain-mediated protein oligomerization and function. Per–ARNT–Sim (PAS) domains are essential modules of many multi-domain signalling proteins that mediate protein interaction and/or sense environmental stimuli. Frequently, multiple PAS domains are present within single polypeptide chains, where their interplay is required for protein function. Although many isolated PAS domain structures have been reported over the last decades, only a few structures of multi-PAS proteins are known. Therefore, the molecular mechanism of multi-PAS domain-mediated protein oligomerization and function is poorly understood. The transcription factor PpsR from Rhodobacter sphaeroides is such a multi-PAS domain protein that, in addition to its three PAS domains, contains a glutamine-rich linker and a C-terminal helix–turn–helix DNA-binding motif. Here, crystal structures of two N-terminally and C-terminally truncated PpsR variants that comprise a single (PpsR{sub Q-PAS1}) and two (PpsR{sub N-Q-PAS1}) PAS domains, respectively, are presented and the multi-step strategy required for the phasing of a triple PAS domain construct (PpsR{sub ΔHTH}) is illustrated. While parts of the biologically relevant dimerization interface can already be observed in the two shorter constructs, the PpsR{sub ΔHTH} structure reveals how three PAS domains enable the formation of multiple oligomeric states (dimer, tetramer and octamer), highlighting that not only the PAS cores but also their α-helical extensions are essential for protein oligomerization. The results demonstrate that the long helical glutamine-rich linker of PpsR results from a direct fusion of the N-cap of the PAS1 domain with the C-terminal extension of the N-domain that

  4. Silver staining of proteins in polyacrylamide gels

    PubMed Central

    Chevallet, Mireille; Luche, Sylvie; Rabilloud, Thierry

    2006-01-01

    Silver staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. It combines excellent sensitivity (in the low nanogram range) whilst using very simple and cheap equipment and chemicals. It is compatible with downstream processing such as mass spectrometry analysis after protein digestion. The sequential phases of silver staining are protein fixation, then sensitization, then silver impregnation and finally image development. Several variants of silver staining are described here, which can be completed in a time range from 2 hours to one day after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks PMID:17487168

  5. Standardization of the Papanicolaou stain. I. A comparison of five nuclear stains.

    PubMed

    Schulte, E; Wittekind, D

    1990-06-01

    The staining characteristics of five nuclear stains used in a Papanicolaou staining procedure were investigated. Alcohol-fixed cervical smears were stained with a modified Papanicolaou procedure using hematoxylin, alcoholic thionin bromide, alcoholic Victoria blue B, gallocyanin or the thionin Feulgen reagent (thionin-SO2) as the nuclear stain. The same anionic counterstain was used for all slides, and the optical densities of cell nuclei and cytoplasm were measured with the IBAS 2000 image analyzer. Alcoholic thionin gave the most intense nuclear stain, with a very high reproducibility of the staining pattern. Hematoxylin showed the highest coefficient of variation of the staining intensity. Both hematoxylin and gallocyanin gave some nonspecific cytoplasmic staining. Thionin-SO2 allowed a quantitative assessment of DNA, but gave a low staining intensity. Staining with the metal complex dyes interfered with subsequent staining with the pararosaniline Feulgen reagent. Alcoholic thioinin is thus recommended as a nuclear stain for cervical cytology in the Papanicolaou procedure, both for image analysis and for visual microscopy. PMID:1695100

  6. A differential staining technique for vertebrate histology.

    PubMed

    Bhattacharyya, T K

    1979-03-01

    A staining method is described for studying micro-anatomy of different vertebrate tissues in the light microscope. A staining sequence of celestin blue--erythrosin--orange G--fast green with mordanting in phosphomolybdic acid yields a satisfactory differentiation and fine colour contrast in various tissues. The efficacy of the method was tested on different avian and mammalian tissues. PMID:86938

  7. Efficiency of staining hair with indocyanine green

    NASA Astrophysics Data System (ADS)

    Kulyabina, Tatyana V.; Kochubey, Vyacheslav I.

    2005-06-01

    The efficiency of staining hair with indocyanine green (ICG) solution depending on type of hair, natural color, staining time and other parameters was investigated. Bonding ICG with hair material occurs due to interaction between ICG molecules and keratinocyte albumin. The penetration of ICG dye into hair meets with difficulties owing to surface protective layer.

  8. Immunofluorescent Staining of Mouse Intestinal Stem Cells

    PubMed Central

    O’Rourke, Kevin P.; Dow, Lukas E; Lowe, Scott W

    2016-01-01

    Immunofluorescent staining of organoids can be performed to visualize molecular markers of cell behavior. For example, cell proliferation marked by incorporation of nucleotide (EdU), or to observe markers of intestinal differentiation including paneth cells, goblet cells, or enterocytes (see Figure 1). In this protocol we detail a method to fix, permeabilize, stain and mount intestinal organoids for analysis by immunofluorescent confocal microscopy.

  9. Faux Pas Detection and Intentional Action in Asperger Syndrome. A Replication on a French Sample

    ERIC Educational Resources Information Center

    Zalla, Tiziana; Sav, Anca-Maria; Stopin, Astrid; Ahade, Sabrina; Leboyer, Marion

    2009-01-01

    In the present study, we investigated mindreading abilities in a group of adults with Asperger Syndrome (AS) by using the faux pas task, an advanced test of theory of mind (Baron-Cohen et al. (1999). "Journal of Autism and Developmental Disorders, 29," 407-418). The faux pas is a particular case of a non-intentional action reflecting an…

  10. Ligand binding PAS domains in a genomic, cellular, and structural context

    PubMed Central

    Henry, Jonathan T.; Crosson, Sean

    2012-01-01

    Per-Arnt-Sim (PAS) domains occur in proteins from all kingdoms of life. In the bacterial kingdom, PAS domains are commonly positioned at the amino terminus of signaling proteins such as sensor histidine kinases, cyclic-di-GMP synthases/hydrolases, and methyl-accepting chemotaxis proteins. Although these domains are highly divergent at the primary sequence level, the structures of dozens of PAS domains across a broad section of sequence space have been solved, revealing a conserved three-dimensional architecture. An all-versus-all alignment of 63 PAS structures demonstrates that the PAS domain family forms structural clades on the basis of two principal variables: (a) topological location inside or outside the plasma membrane and (b) the class of small molecule that they bind. The binding of a chemically diverse range of small-molecule metabolites is a hallmark of the PAS domain family. PAS ligand binding either functions as a primary cue to initiate a cellular signaling response or provides the domain with the capacity to respond to secondary physical or chemical signals such as gas molecules, redox potential, or photons. This review synthesizes the current state of knowledge of the structural foundations and evolution of ligand recognition and binding by PAS domains. PMID:21663441

  11. Reductive evolution and the loss of PDC/PAS domains from the genus Staphylococcus

    PubMed Central

    2013-01-01

    Background The Per-Arnt-Sim (PAS) domain represents a ubiquitous structural fold that is involved in bacterial sensing and adaptation systems, including several virulence related functions. Although PAS domains and the subclass of PhoQ-DcuS-CitA (PDC) domains have a common structure, there is limited amino acid sequence similarity. To gain greater insight into the evolution of PDC/PAS domains present in the bacterial kingdom and staphylococci in specific, the PDC/PAS domains from the genomic sequences of 48 bacteria, representing 5 phyla, were identified using the sensitive search method based on HMM-to-HMM comparisons (HHblits). Results A total of 1,007 PAS domains and 686 PDC domains distributed over 1,174 proteins were identified. For 28 Gram-positive bacteria, the distribution, organization, and molecular evolution of PDC/PAS domains were analyzed in greater detail, with a special emphasis on the genus Staphylococcus. Compared to other bacteria the staphylococci have relatively fewer proteins (6–9) containing PDC/PAS domains. As a general rule, the staphylococcal genomes examined in this study contain a core group of seven PDC/PAS domain-containing proteins consisting of WalK, SrrB, PhoR, ArlS, HssS, NreB, and GdpP. The exceptions to this rule are: 1) S. saprophyticus lacks the core NreB protein; 2) S. carnosus has two additional PAS domain containing proteins; 3) S. epidermidis, S. aureus, and S. pseudintermedius have an additional protein with two PDC domains that is predicted to code for a sensor histidine kinase; 4) S. lugdunensis has an additional PDC containing protein predicted to be a sensor histidine kinase. Conclusions This comprehensive analysis demonstrates that variation in PDC/PAS domains among bacteria has limited correlations to the genome size or pathogenicity; however, our analysis established that bacteria having a motile phase in their life cycle have significantly more PDC/PAS-containing proteins. In addition, our analysis revealed a

  12. Role of the PAS1 gene of Pichia pastoris in peroxisome biogenesis

    PubMed Central

    1994-01-01

    Several groups have reported the cloning and sequencing of genes involved in the biogenesis of yeast peroxisomes. Yeast strains bearing mutations in these genes are unable to grow on carbon sources whose metabolism requires peroxisomes, and these strains lack morphologically normal peroxisomes. We report the cloning of Pichia pastoris PAS1, the homologue (based on a high level of protein sequence similarity) of the Saccharomyces cerevisiae PAS1. We also describe the creation and characterization of P. pastoris pas1 strains. Electron microscopy on the P. pastoris pas1 cells revealed that they lack morphologically normal peroxisomes, and instead contain membrane-bound structures that appear to be small, mutant peroxisomes, or "peroxisome ghosts." These "ghosts" proliferated in response to induction on peroxisome-requiring carbon sources (oleic acid and methanol), and they were distributed to daughter cells. Biochemical analysis of cell lysates revealed that peroxisomal proteins are induced normally in pas1 cells. Peroxisome ghosts from pas1 cells were purified on sucrose gradients, and biochemical analysis showed that these ghosts, while lacking several peroxisomal proteins, did import varying amounts of several other peroxisomal proteins. The existence of detectable peroxisome ghosts in P. pastoris pas1 cells, and their ability to import some proteins, stands in contrast with the results reported by Erdmann et al. (1991) for the S. cerevisiae pas1 mutant, in which they were unable to detect peroxisome-like structures. We discuss the role of PAS1 in peroxisome biogenesis in light of the new information regarding peroxisome ghosts in pas1 cells. PMID:7962088

  13. Compact, Automated Centrifugal Slide-Staining System

    NASA Technical Reports Server (NTRS)

    Feeback, Daniel L.; Clarke, Mark S. F.

    2004-01-01

    The Directional Acceleration Vector-Driven Displacement of Fluids (DAVD-DOF) system, under development at the time of reporting the information for this article, would be a relatively compact, automated, centrifugally actuated system for staining blood smears and other microbiological samples on glass microscope slides in either a microgravitational or a normal Earth gravitational environment. The DAVD-DOF concept is a successor to the centrifuge-operated slide stainer (COSS) concept, which was reported in Slide-Staining System for Microgravity or Gravity (MSC-22949), NASA Tech Briefs, Vol. 25, No. 1 (January, 2001), page 64. The COSS includes reservoirs and a staining chamber that contains a microscope slide to which a biological sample is affixed. The staining chamber is sequentially filled with and drained of staining and related liquids from the reservoirs by use of a weighted plunger to force liquid from one reservoir to another at a constant level of hypergravity maintained in a standard swing-bucket centrifuge. In the DAVD-DOF system, a staining chamber containing a sample would also be sequentially filled and emptied, but with important differences. Instead of a simple microscope slide, one would use a special microscope slide on which would be fabricated a network of very small reservoirs and narrow channels connected to a staining chamber (see figure). Unlike in the COSS, displacement of liquid would be effected by use of the weight of the liquid itself, rather than the weight of a plunger.

  14. Multicenter Assessment of Gram Stain Error Rates.

    PubMed

    Samuel, Linoj P; Balada-Llasat, Joan-Miquel; Harrington, Amanda; Cavagnolo, Robert

    2016-06-01

    Gram stains remain the cornerstone of diagnostic testing in the microbiology laboratory for the guidance of empirical treatment prior to availability of culture results. Incorrectly interpreted Gram stains may adversely impact patient care, and yet there are no comprehensive studies that have evaluated the reliability of the technique and there are no established standards for performance. In this study, clinical microbiology laboratories at four major tertiary medical care centers evaluated Gram stain error rates across all nonblood specimen types by using standardized criteria. The study focused on several factors that primarily contribute to errors in the process, including poor specimen quality, smear preparation, and interpretation of the smears. The number of specimens during the evaluation period ranged from 976 to 1,864 specimens per site, and there were a total of 6,115 specimens. Gram stain results were discrepant from culture for 5% of all specimens. Fifty-eight percent of discrepant results were specimens with no organisms reported on Gram stain but significant growth on culture, while 42% of discrepant results had reported organisms on Gram stain that were not recovered in culture. Upon review of available slides, 24% (63/263) of discrepant results were due to reader error, which varied significantly based on site (9% to 45%). The Gram stain error rate also varied between sites, ranging from 0.4% to 2.7%. The data demonstrate a significant variability between laboratories in Gram stain performance and affirm the need for ongoing quality assessment by laboratories. Standardized monitoring of Gram stains is an essential quality control tool for laboratories and is necessary for the establishment of a quality benchmark across laboratories. PMID:26888900

  15. Postural Assessment Software (PAS/SAPO): Validation and Reliabiliy

    PubMed Central

    Ferreira, Elizabeth Alves G.; Duarte, Marcos; Maldonado, Edison Puig; Burke, Thomaz Nogueira; Marques, Amelia Pasqual

    2010-01-01

    OBJECTIVE: This study was designed to estimate the accuracy of the postural assessment software (PAS/SAPO) for measurement of corporal angles and distances as well as the inter- and intra-rater reliabilities. INTRODUCTION: Postural assessment software was developed as a subsidiary tool for postural assessment. It is easy to use and available in the public domain. Nonetheless, validation studies are lacking. METHODS: The study sample consisted of 88 pictures from 22 subjects, and each subject was assessed twice (1 week interval) by 5 blinded raters. Inter- and intra-rater reliabilities were estimated using the intraclass correlation coefficient. To estimate the accuracy of the software, an inanimate object was marked with hallmarks using pre-established parameters. Pictures of the object were rated, and values were checked against the known parameters. RESULTS: Inter-rater reliability was excellent for 41% of the variables and very good for 35%. Ten percent of the variables had acceptable reliability, and 14% were defined as non-acceptable. For intra-rater reliability, 44.8% of the measurements were considered to be excellent, 23.5% were very good, 12.4% were acceptable and 19.3% were considered non-acceptable. Angular measurements had a mean error analisys of 0.11°, and the mean error analisys for distance was 1.8 mm. DISCUSSION: Unacceptable intraclass correlation coefficient values typically used the vertical line as a reference, and this may have increased the inaccuracy of the estimates. Increased accuracies were obtained by younger raters with more sophisticated computer skills, suggesting that past experience influenced results. CONCLUSION: The postural assessment software was accurate for measuring corporal angles and distances and should be considered as a reliable tool for postural assessment. PMID:20668624

  16. Gram staining apparatus for space station applications

    NASA Technical Reports Server (NTRS)

    Molina, T. C.; Brown, H. D.; Irbe, R. M.; Pierson, D. L.

    1990-01-01

    A self-contained, portable Gram staining apparatus (GSA) has been developed for use in the microgravity environment on board the Space Station Freedom. Accuracy and reproducibility of this apparatus compared with the conventional Gram staining method were evaluated by using gram-negative and gram-positive controls and different species of bacteria grown in pure cultures. A subsequent study was designed to assess the performance of the GSA with actual specimens. A set of 60 human and environmental specimens was evaluated with the GSA and the conventional Gram staining procedure. Data obtained from these studies indicated that the GSA will provide the Gram staining capability needed for the microgravity environment of space.

  17. An alternative to India ink stain.

    PubMed

    Ibembe, Isaac Nicholas; Wiggin, Timothy Roger

    2015-07-01

    Accessing India ink in rural Uganda is difficult and costly. An alternative stain was sought to assist in microbiological diagnoses of cryptococcal infections in immunosuppressed patients with meningitis. Mascara proved to be an excellent and cheap alternative. PMID:25999353

  18. Gram staining apparatus for space station applications.

    PubMed Central

    Molina, T C; Brown, H D; Irbe, R M; Pierson, D L

    1990-01-01

    A self-contained, portable Gram staining apparatus (GSA) has been developed for use in the microgravity environment on board the Space Station Freedom. Accuracy and reproducibility of this apparatus compared with the conventional Gram staining method were evaluated by using gram-negative and gram-positive controls and different species of bacteria grown in pure cultures. A subsequent study was designed to assess the performance of the GSA with actual specimens. A set of 60 human and environmental specimens was evaluated with the GSA and the conventional Gram staining procedure. Data obtained from these studies indicated that the GSA will provide the Gram staining capability needed for the microgravity environment of space. Images PMID:1690529

  19. New Grocott Stain without Using Chromic Acid

    PubMed Central

    Shiogama, Kazuya; Kitazawa, Kayo; Mizutani, Yasuyoshi; Onouchi, Takanori; Inada, Ken-ichi; Tsutsumi, Yutaka

    2015-01-01

    We established a new “ecological” Grocott stain for demonstrating fungi, based upon a 4R principle of refusal, reduction, reuse, and recycle of waste management. Conventional Grocott stain employs environmentally harsh 5% chromic acid for oxidization. Initially, we succeeded in reducing the concentration of chromic acid from 5% to 1% by incubating the solution at 60°C and using five-fold diluted chromic acid solution at which point it was reusable. Eventually, we reached the refusal level where 1% periodic acid oxidization was efficient enough, when combined with preheating of sections in the electric jar, microwave oven, or pressure pan. For convenience sake, we recommend pressure pan heating in tap water for 10 min. Stainability of fungi in candidiasis and aspergillosis was comparable with conventional Grocott stain, while Mucor hyphae showed enhanced staining. The modified sequence was further applicable to detecting a variety of mycotic pathogens in paraffin sections. Our environmentally-friendly Grocott stain also has the advantage of avoiding risk of human exposure to hexavalent chromium solution in the histopathology laboratory. The simple stain sequence is can be easily applied worldwide. PMID:25861133

  20. Changes at the KinA PAS-A Dimerization Interface Influence Histidine Kinase Function

    SciTech Connect

    Lee, James; Tomchick, Diana R.; Brautigam, Chad A.; Machius, Mischa; Kort, Remco; Hellingwerf, Klaas J.; Gardner, Kevin H.

    2008-11-12

    The Bacillus subtilis KinA protein is a histidine protein kinase that controls the commitment of this organism to sporulate in response to nutrient deprivation and several other conditions. Prior studies indicated that the N-terminal Per-ARNT-Sim domain (PAS-A) plays a critical role in the catalytic activity of this enzyme, as demonstrated by the significant decrease of the autophosphorylation rate of a KinA protein lacking this domain. On the basis of the environmental sensing role played by PAS domains in a wide range of proteins, including other bacterial sensor kinases, it has been suggested that the PAS-A domain plays an important regulatory role in KinA function. We have investigated this potential by using a combination of biophysical and biochemical methods to examine PAS-A structure and function, both in isolation and within the intact protein. Here, we present the X-ray crystal structure of the KinA PAS-A domain, showing that it crystallizes as a homodimer using {beta}-sheet/{beta}-sheet packing interactions as observed for several other PAS domain complexes. Notably, we observed two dimers with tertiary and quaternary structure differences in the crystalline lattice, indicating significant structural flexibility in these domains. To confirm that KinA PAS-A also forms dimers in solution, we used a combination of NMR spectroscopy, gel filtration chromatography, and analytical ultracentrifugation, the results of which are all consistent with the crystallographic results. We experimentally tested the importance of several residues at the dimer interface using site-directed mutagenesis, finding changes in the PAS-A domain that significantly alter KinA enzymatic activity in vitro and in vivo. These results support the importance of PAS domains within KinA and other histidine kinases and suggest possible routes for natural or artificial regulation of kinase activity.

  1. Digital stain separation for histological images.

    PubMed

    Tadrous, P J

    2010-11-01

    It is often desirable to perform digital image analyses on sections prepared for human interpretation, e.g. nuclear chromatin texture analysis or three-dimensional reconstructions using sections requiring human delineation of structures of interest. Unfortunately such analyses are often more effective using stains with less complex contrast. Here an automated selective 'de-staining' method for digital images is presented. The method separates an image into its red, green and blue and hue, saturation and intensity components. A mask of stained tissue is prepared by automatic percentile thresholding. A single weighted inverted colour channel is then added to each of the three primary colour channels separately by an iterative algorithm that adjusts the weights to give minimum variance within the mask. The modified red, green and blue channels are then recombined. This method is automatic requiring no pre-definition of stain colours or special hardware. The method is demonstrated to 'de-stain' nuclei in haematoxylin and eosin (H&E) sections (and a separate haematoxylin image can be derived from this). An image of isolated brown reaction product is produced with immunoperoxidase preparations counterstained with haematoxylin. Furthermore trichrome (haematoxylin van Gieson, picrosirius red) and other common stains may be separated into their components with modifications of the same algorithm. Although other methods for colour separation do exist (e.g. spectral pathology and colour deconvolution) these require special apparatus or precise calibration and foreknowledge of pure dye colour spectra. The present method of digital stain separation is fully automatic with no such prerequisites. PMID:20946383

  2. Protein Dynamics of the HIF-2α PAS-B Domain upon Heterodimerization and Ligand Binding

    PubMed Central

    Masetti, Matteo; Falchi, Federico; Recanatini, Maurizio

    2014-01-01

    Hypoxia-Inducible Factor (HIF) transcription factors are heterodimeric proteins involved in the regulation of oxygen homeostatis. Their upregulation has been related to several tumors with a remarkably poor clinical outcome. The recent discovery of a druggable cavity in the HIF-2α PAS-B domain has opened an unprecedented opportunity for targeting the HIF-2α transcription factor in view of pharmaceutical strategies. Coincidentally, a novel compound able to selectively disrupt the HIF heterodimerization with a submicromolar activity has been reported. In this work, we investigated the molecular mechanisms responsible for the inhibition by comparing the dynamical features of the HIF-2α PAS-B monomer and the HIF-2α PAS-B/HIF-1β PAS-B complex, in the ligand-bound and -unbound states. Plain and biased Molecular Dynamics were used to characterize the differential conformational changes both structurally and energetically. PMID:24736273

  3. Toronto Psoriatic Arthritis Screening (ToPAS) questionnaire: a report from the GRAPPA 2009 annual meeting.

    PubMed

    Chandran, Vinod; Gladman, Dafna D

    2011-03-01

    The Toronto Psoriatic Arthritis Screening questionnaire (ToPAS) was developed as a tool to screen for psoriatic arthritis (PsA) in patients with psoriasis as well as in the general population. Thus, it differs from PsA-specific screening tools and may be used to screen for PsA in epidemiologic and family investigations. In a presentation at the 2009 annual meeting of GRAPPA (Group for Research and Assessment of Psoriasis and Psoriatic Arthritis) in Stockholm, Sweden, the authors described the development, testing, and validation of the ToPAS tool. Results of a comparison of the ToPAS questionnaire with the Psoriatic Arthritis Screening and Evaluation (PASE) questionnaire were also presented. Modification and further validation of the ToPAS are under way. PMID:21362783

  4. Third Annual Maintenance Inspection & Test Report for PAS-1 Cask Certification for Shipping Payload B

    SciTech Connect

    KELLY, D.L.

    2001-11-01

    This, Third Annual Maintenance, Inspection, and Test Report for PAS-1 Cask Certification for Shipping Payload B, documents the successful completion of maintenance, testing, and inspections for two casks, serial numbers 2161-026 and 2162-027.

  5. J-PAS : Low-resolution (R ~ 50) spectroscopy covering 8000 deg2

    NASA Astrophysics Data System (ADS)

    López-Sanjuan, C.; Cenarro, A. J.; Díaz-García, L. A.; Muniesa, D. J.; San Roman, I.; Varela, J.; Viironen, K.

    2015-02-01

    We present the ambitious project J-PAS, that will cover 8000 deg2 of the northern sky with 54 narrow-band (~145Å) contiguous filters, all of them in the optical range (3700Å-9200Å). J-PAS will provide a low resolution spectra (R ~ 50) in every pixel of the northern sky by 2020, leading to excellent photometric redshifts (0.3% uncertainty) of 100 million sources. J-PAS will permit the study of the 2D properties of nearby galaxies with unprecedented statistics. Some viable studies are the distribution of the star formation rate traced by Hα, the stellar populations gradients in elliptical galaxies up to a few effective radii, or the impact of environment in galaxy properties. In summary, J-PAS will bring a superb data set for 3D analysis in the local Universe.

  6. Compositions for chromosome-specific staining

    DOEpatents

    Gray, J.W.; Pinkel, D.

    1998-05-26

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. The methods produce staining patterns that can be tailored for specific cytogenetic analyses. The probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. The invention provides for automated means to detect and analyze chromosomal abnormalities. 17 figs.

  7. Compositions for chromosome-specific staining

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel

    1998-01-01

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.

  8. Changes in Quaternary Structure in the Signaling Mechanisms of PAS Domains

    SciTech Connect

    Ayers, Rebecca A.; Moffat, Keith

    2008-12-15

    FixL from Bradyrhizobium japonicum is a PAS sensor protein in which two PAS domains covalently linked to a histidine kinase domain are responsible for regulating nitrogen fixation in an oxygen-dependent manner. The more C-terminal PAS domain, denoted bjFixLH, contains a heme cofactor that binds diatomic molecules such as carbon monoxide and oxygen and regulates the activity of the FixL histidine kinase as part of a two-component signaling system. We present the structures of ferric, deoxy, and carbon monoxide-bound bjFixLH in a new space group (P1) and at resolutions (1.5--1.8 {angstrom}) higher than the resolutions of those previously obtained. Interestingly, bjFixLH can form two different dimers (in P1 and R32 crystal forms) in the same crystallization solution, where the monomers in one dimer are rotated {approx}175 deg. relative to the second. This suggests that PAS monomers are plastic and that two quite distinct quaternary structures are closely similar in free energy. We use screw rotation analysis to carry out a quantitative pairwise comparison of PAS quaternary structures, which identifies five different relative orientations adopted by isolated PAS monomers. We conclude that PAS monomer arrangement is context-dependent and could differ depending on whether the PAS domains are isolated or are part of a full-length protein. Structurally homologous residues comprise a conserved dimer interface. Using network analysis, we find that the architecture of the PAS dimer interface is continuous rather than modular; the network of residues comprising the interface is strongly connected. A continuous dimer interface is consistent with the low dimer-monomer dissociation equilibrium constant. Finally, we quantitate quaternary structural changes induced by carbon monoxide binding to a bjFixLH dimer, in which monomers rotate by up to 2 deg. relative to each other. We relate these changes to those in other dimeric PAS domains and discuss the role of quaternary structural

  9. Changes in quaternary structure in the signaling mechanisms of PAS domains.

    PubMed

    Ayers, Rebecca A; Moffat, Keith

    2008-11-18

    FixL from Bradyrhizobium japonicum is a PAS sensor protein in which two PAS domains covalently linked to a histidine kinase domain are responsible for regulating nitrogen fixation in an oxygen-dependent manner. The more C-terminal PAS domain, denoted bjFixLH, contains a heme cofactor that binds diatomic molecules such as carbon monoxide and oxygen and regulates the activity of the FixL histidine kinase as part of a two-component signaling system. We present the structures of ferric, deoxy, and carbon monoxide-bound bjFixLH in a new space group ( P1) and at resolutions (1.5-1.8 A) higher than the resolutions of those previously obtained. Interestingly, bjFixLH can form two different dimers (in P1 and R32 crystal forms) in the same crystallization solution, where the monomers in one dimer are rotated approximately 175 degrees relative to the second. This suggests that PAS monomers are plastic and that two quite distinct quaternary structures are closely similar in free energy. We use screw rotation analysis to carry out a quantitative pairwise comparison of PAS quaternary structures, which identifies five different relative orientations adopted by isolated PAS monomers. We conclude that PAS monomer arrangement is context-dependent and could differ depending on whether the PAS domains are isolated or are part of a full-length protein. Structurally homologous residues comprise a conserved dimer interface. Using network analysis, we find that the architecture of the PAS dimer interface is continuous rather than modular; the network of residues comprising the interface is strongly connected. A continuous dimer interface is consistent with the low dimer-monomer dissociation equilibrium constant. Finally, we quantitate quaternary structural changes induced by carbon monoxide binding to a bjFixLH dimer, in which monomers rotate by up to approximately 2 degrees relative to each other. We relate these changes to those in other dimeric PAS domains and discuss the role of

  10. PAS-cal: a Generic Recombinant Peptide Calibration Standard for Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Breibeck, Joscha; Serafin, Adam; Reichert, Andreas; Maier, Stefan; Küster, Bernhard; Skerra, Arne

    2014-08-01

    We describe the design, preparation, and mass-spectrometric characterization of a new recombinant peptide calibration standard with uniform biophysical and ionization characteristics for mass spectrometry. "PAS-cal" is an artificial polypeptide concatamer of peptide cassettes with varying lengths, each composed of the three small, chemically stable amino acids Pro, Ala, and Ser, which are interspersed by Arg residues to allow site-specific cleavage with trypsin. PAS-cal is expressed at high yields in Escherichia coli as a Small Ubiquitin-like MOdifier (SUMO) fusion protein, which is easily purified and allows isolation of the PAS-cal moiety after SUMO protease cleavage. Upon subsequent in situ treatment with trypsin, the PAS-cal polypeptide yields a set of four defined homogeneous peptides in the range from 2 to 8 kDa with equal mass spacing. ESI-MS analysis revealed a conveniently interpretable raw spectrum, which after deconvolution resulted in a very simple pattern of four peaks with similar ionization signals. MALDI-MS analysis of a PAS-cal peptide mixture comprising both the intact polypeptide and its tryptic fragments revealed not only the four standard peptides but also the singly and doubly charged states of the intact concatamer as well as di- and trimeric adduct ion species between the peptides, thus augmenting the observable m/z range. The advantageous properties of PAS-cal are most likely a result of the strongly hydrophilic and conformationally disordered PEG-like properties of the PAS sequences. Therefore, PAS-cal offers an inexpensive and versatile recombinant peptide calibration standard for mass spectrometry in protein/peptide bioanalytics and proteomics research, the composition of which may be further adapted to fit individual needs.

  11. Subclinical Onychomycosis in Patients With Type II Diabetes

    PubMed Central

    El Tawdy, Amira; Zaki, Naglaa; Alfishawy, Mostafa; Rateb, Amr

    2015-01-01

    Fungal organisms could be present in the nail without any clinical manifestations. As onychomycosis in diabetics has more serious complications, early detection of such infection could be helpful to prevent them. We aim in this study to assess the possibility of detecting subclinical onychomycosis in type II diabetic patients and addressing possible associated neuropathy. A cross sectional, observational study included patients with type II diabetes with normal big toe nail. All were subjected to nail clipping of the big toe nail, followed by staining with Hematoxylin and Eosin and Periodic-Acid-Schiff (PAS) stains and examined microscopically. A total of 106 patients were included, fungal infection was identified in eight specimens, all were uncontrolled diabetes, and six had neuropathy. Using the nail clipping and microscopic examination with PAS stain to detect such subclinical infection could be an applicable screening test for diabetic patients, for early detection and management of onychomycosis. PMID:26734120

  12. Detection Of Concrete Deterioration By Staining

    DOEpatents

    Guthrie, Jr., George D.; Carey, J. William

    1999-09-21

    A method using concentrated aqueous solutions of sodium cobaltinitrite and a rhodamine dye is described which can be used to identify concrete that contains gels formed by the alkali-silica reaction (ASR), and to identify degraded concrete which results in a porous or semi-permeable paste due to carbonation or leaching. These solutions present little health or environmental risk, are readily applied, and rapidly discriminate between two chemically distinct gels; K-rich, Na--K--Ca--Si gels are identified by yellow staining, and alkali-poor, Ca--Si gels are identified by pink staining.

  13. Laser treatment of port-wine stains

    PubMed Central

    Brightman, Lori A; Geronemus, Roy G; Reddy, Kavitha K

    2015-01-01

    Port-wine stains are a type of capillary malformation affecting 0.3% to 0.5% of the population. Port-wine stains present at birth as pink to erythematous patches on the skin and/or mucosa. Without treatment, the patches typically darken with age and may eventually develop nodular thickening or associated pyogenic granuloma. Laser and light treatments provide improvement through selective destruction of vasculature. A variety of vascular-selective lasers may be employed, with the pulsed dye laser being the most common and well studied. Early treatment produces more optimal results. Advances in imaging and laser treatment technologies demonstrate potential to further improve clinical outcomes. PMID:25624768

  14. Automated single-slide staining device

    NASA Technical Reports Server (NTRS)

    Wilkins, J. R.; Mills, S. M. (Inventor)

    1977-01-01

    A simple apparatus and method is disclosed for making individual single Gram stains on bacteria inoculated slides to assist in classifying bacteria in the laboratory as Gram-positive or Gram-negative. The apparatus involves positioning a single inoculated slide in a stationary position and thereafter automatically and sequentially flooding the slide with increments of a primary stain, a mordant, a decolorizer, a counterstain and a wash solution in a sequential manner without the individual lab technician touching the slide and with minimum danger of contamination thereof from other slides.

  15. Cache Domains That are Homologous to, but Different from PAS Domains Comprise the Largest Superfamily of Extracellular Sensors in Prokaryotes

    PubMed Central

    Upadhyay, Amit A.; Fleetwood, Aaron D.; Adebali, Ogun; Finn, Robert D.; Zhulin, Igor B.

    2016-01-01

    Cellular receptors usually contain a designated sensory domain that recognizes the signal. Per/Arnt/Sim (PAS) domains are ubiquitous sensors in thousands of species ranging from bacteria to humans. Although PAS domains were described as intracellular sensors, recent structural studies revealed PAS-like domains in extracytoplasmic regions in several transmembrane receptors. However, these structurally defined extracellular PAS-like domains do not match sequence-derived PAS domain models, and thus their distribution across the genomic landscape remains largely unknown. Here we show that structurally defined extracellular PAS-like domains belong to the Cache superfamily, which is homologous to, but distinct from the PAS superfamily. Our newly built computational models enabled identification of Cache domains in tens of thousands of signal transduction proteins including those from important pathogens and model organisms. Furthermore, we show that Cache domains comprise the dominant mode of extracellular sensing in prokaryotes. PMID:27049771

  16. Pleural and Pulmonary Staining at Inferior Phrenic Arteriography Mimicking a Tumor Staining of Hepatocellular Carcinoma

    SciTech Connect

    Lee, Deok Hee; Hwang, Jae Cheol; Lim, Soo Mee; Yoon, Hyun-Ki; Sung, Kyu-Bo; Song, Ho-Young

    2000-03-15

    Purpose: To describe the findings of pleural and pulmonary staining of the inferior phrenic artery, which can be confused with tumor staining during transarterial chemoembolization (TACE) of hepatoma.Methods: Fifteen patients who showed pleural and pulmonary staining without relationship to hepatic masses at inferior phrenic arteriography were enrolled. The staining was noted at initial TACE (n = 8), at successive TACE (n = 5), and after hepatic surgery (n = 2). The angiographic pattern, the presence of pleural change on computed tomography (CT), and clinical history were evaluated.Results: Draining pulmonary veins were seen in all cases. The lower margin of the staining corresponded to the lower margin of the pleura in 10 patients. CT showed pleural and/or pulmonary abnormalities in all cases. After embolization of the inferior phrenic artery, the accumulation of iodized oil in the lung was noted.Conclusion: Understanding the CT and angiographic findings of pleural and pulmonary staining during TACE may help differentiate benign staining from tumor staining.

  17. The Language of Stained-Glass Windows

    ERIC Educational Resources Information Center

    Brew, Charl Anne

    2010-01-01

    The splendor and beauty of stained glass punctuates any room. In this article, the author describes a cross-curriculum project which incorporated the French classes' research and written study of France in the Middle Ages. For the project the author suggested Sainte-Chapelle which is considered a reliquary and was built by Louis IX to house the…

  18. Protein Stains to Detect Antigen on Membranes.

    PubMed

    Dsouza, Anil; Scofield, R Hal

    2015-01-01

    Western blotting (protein blotting/electroblotting) is the gold standard in the analysis of complex protein mixtures. Electroblotting drives protein molecules from a polyacrylamide (or less commonly, of an agarose) gel to the surface of a binding membrane, thereby facilitating an increased availability of the sites with affinity for both general and specific protein reagents. The analysis of these complex protein mixtures is achieved by the detection of specific protein bands on a membrane, which in turn is made possible by the visualization of protein bands either by chemical staining or by reaction with an antibody of a conjugated ligand. Chemical methods employ staining with organic dyes, metal chelates, autoradiography, fluorescent dyes, complexing with silver, or prelabeling with fluorophores. All of these methods have differing sensitivities and quantitative determinations vary significantly. This review will describe the various protein staining methods applied to membranes after western blotting. "Detection" precedes and is a prerequisite to obtaining qualitative and quantitative data on the proteins in a sample, as much as to comparing the protein composition of different samples. "Detection" is often synonymous to staining, i.e., the reversible or irreversible binding by the proteins of a colored organic or inorganic chemical. PMID:26139252

  19. Method for copper staining of germanium crystals

    NASA Technical Reports Server (NTRS)

    Rivet, E. J.

    1969-01-01

    Proper conditions for copper staining of germanium crystals include a low solution temperature of 3 degrees C, illumination of the sample by infrared light, and careful positioning of the light source relative to the sample so as to minimize absorption of the infrared light.

  20. Asbestos identification by dispersion staining microscopy.

    PubMed

    Ganotes, J T; Tan, H T

    1980-01-01

    Asbestos can be detected and identified by an optical microscope procedure known as dispersion staining. This procedure can be carried out with most phase contrast equipped microscopes. The primary application is for material samples. Distinction between tremolite and anthophyllite asbestos requires examination between crossed polarizers. PMID:6153496

  1. Evaluating the PAS-SIM model using a passive air sampler calibration study for pesticides.

    PubMed

    Restrepo, Andrés Ramírez; Hayward, Stephen J; Armitage, James M; Wania, Frank

    2015-07-01

    The main objective of this study was to evaluate the performance of a model for simulating the uptake of various pesticides on passive air samplers (PAS). From 2006-2007 a series of PAS using XAD-resin were deployed at Egbert, a rural agricultural site in southern Ontario, Canada, to measure the uptake of pesticides for time periods ranging from two months to one year. A continuous increase in sequestered amounts was observed for most pesticides, except for trifluralin and pendimethalin, which could conceivably be subject to substantial degradation inside the sampler. Continuous low-volume active air samples taken during the same period, along with data on weather conditions, allowed for the simulation of the uptake of the pesticides using the model (PAS-SIM). The modelled accumulation of pesticides on the PAS over the deployment period was in good agreement with the experimental data in most cases (i.e., within a factor of two) providing insight into the uptake kinetics of this type of sampler in the field. Passive sampling rates (PSR, m(3) d(-1)) were determined from the empirical data generated for this study using three different methods and compared with the PSRs generated by the model. Overall, the PAS-SIM model, which is capable of accounting for the influence of temperature and wind variations on PSRs, provided reasonable results that range between the three empirical approaches employed and well-established literature values. Further evaluation and application of the PAS-SIM model to explore the potential spatial and temporal variability in PAS uptake kinetics is warranted, particularly for established monitoring sites where detailed meteorological data are more likely to be available. PMID:26083201

  2. A method for the staining of intraosseous nerve fibers using Sihler's staining technique.

    PubMed

    Shiozaki, K; Miida, K; Tanaka, R; Shimoda, S

    2013-08-01

    Understanding nerve fiber distribution in the jaw bone is important when performing invasive surgical treatments. Both microscopic and macroscopic anatomical techniques have been developed to study innervation. Conventional methods of removing and staining these structures, however, often alter structure and lack reproducibility of the resulting specimens. We sought to optimize Sihler's staining technique to stain intraosseous nerves in mandibles. Four cadaver specimens were used. The best staining of intraosseous nerve fibers was achieved by using the Plank-Rychlo solution. When the Styrene monomer was used, the resulting transparency was better than that obtained with glycerin under the same conditions. No significant differences were found between Sihler's staining procedure performed according to the conventional method and the procedure in which the second decalcification step was omitted. Our results demonstrate that applying Sihler's staining technique to bones makes them transparent and allows observation of nerves while preserving the external shape of the bone and maintaining the position of intraosseous nerve fibers. Our findings suggest our Sihler staining method for intraosseous nerve fibers can provide an intermediate resolution between macroscopic and microscopic techniques. PMID:23472877

  3. Improved Whole-Blood-Staining Device

    NASA Technical Reports Server (NTRS)

    Sams, Clarence F.; Crucian, Brian; Paul, Bonnie; Melton, Shannon; Guess, Terry

    2012-01-01

    Dramatic improvements have been made in NASA s Whole Blood Staining Device (WBSD) since it was last described in "Whole-Blood-Staining Device," NASA Tech Briefs, Vol. 23, No. 10 (October 1999), page 64. The new system has a longer shelf life, a simpler and more effective operational procedure, improved interface with instrumentation, and shorter processing time. More specifically, the improvements have targeted bag and locking clip materials, sampling ports, and air pocket prevention. The WBSD stains whole blood collected during spaceflight for subsequent flow cytometric analysis. In short, the main device stains white blood cells by use of monoclonal antibodies conjugated to various fluorochromes, followed by lysing and fixing of the cells by use of a commercial reagent that has been diluted according to NASA safety standards. This system is compact, robust, and does not require electric power, precise mixing, or precise incubation times. Figure 1 depicts the present improved version for staining applications, which is a poly(tetrafluoroethylene) bag with a Luer-lock port and plastic locking clips. An InterLink (or equivalent) intravenous- injection port screws into the Luer-lock port. The inflatable/collapsible nature of the bag facilitates loading and helps to minimize the amount of air trapped in the fully loaded bag. Some additional uses have been identified for the device beyond whole blood staining. The WBSD has been configured for functional assays that require culture of live cells by housing sterile culture media, mitogens, and fixatives prior to use [Figure 2(a)]. Simple injection of whole blood allows cell-stimulation culture to be performed in reduced gravity conditions, and product stabilization prior to storage, while protecting astronauts from liquid biohazardous materials. Also, the improved WBSD has reconstituted powdered injectable antibiotics by mixing them with diluent liquids [Figure 2(b)]. Although such mixing can readily be performed on

  4. Complex and dynamic landscape of RNA polyadenylation revealed by PAS-Seq

    PubMed Central

    Shepard, Peter J.; Choi, Eun-A; Lu, Jente; Flanagan, Lisa A.; Hertel, Klemens J.; Shi, Yongsheng

    2011-01-01

    Alternative polyadenylation (APA) of mRNAs has emerged as an important mechanism for post-transcriptional gene regulation in higher eukaryotes. Although microarrays have recently been used to characterize APA globally, they have a number of serious limitations that prevents comprehensive and highly quantitative analysis. To better characterize APA and its regulation, we have developed a deep sequencing-based method called Poly(A) Site Sequencing (PAS-Seq) for quantitatively profiling RNA polyadenylation at the transcriptome level. PAS-Seq not only accurately and comprehensively identifies poly(A) junctions in mRNAs and noncoding RNAs, but also provides quantitative information on the relative abundance of polyadenylated RNAs. PAS-Seq analyses of human and mouse transcriptomes showed that 40%–50% of all expressed genes produce alternatively polyadenylated mRNAs. Furthermore, our study detected evolutionarily conserved polyadenylation of histone mRNAs and revealed novel features of mitochondrial RNA polyadenylation. Finally, PAS-Seq analyses of mouse embryonic stem (ES) cells, neural stem/progenitor (NSP) cells, and neurons not only identified more poly(A) sites than what was found in the entire mouse EST database, but also detected significant changes in the global APA profile that lead to lengthening of 3′ untranslated regions (UTR) in many mRNAs during stem cell differentiation. Together, our PAS-Seq analyses revealed a complex landscape of RNA polyadenylation in mammalian cells and the dynamic regulation of APA during stem cell differentiation. PMID:21343387

  5. Structure and Dimerization Properties of the Aryl Hydrocarbon Receptor PAS-A Domain

    PubMed Central

    Wu, Dalei; Potluri, Nalini; Kim, Youngchang

    2013-01-01

    The aryl hydrocarbon receptor (AHR) is a ligand-dependent transcription factor that binds to xenobiotics and responds by regulating the expression of gene programs required for detoxification and metabolism. AHR and its heterodimerization partner aryl hydrocarbon receptor nuclear translocator (ARNT) belong to the basic helix-loop-helix (bHLH)–PER-ARNT-SIM (PAS) family of transcription factors. Here we report the 2.55-Å-resolution crystal structure of the mouse AHR PAS-A domain, which represents the first AHR-derived protein structure. The AHR PAS-A domain forms a helix-swapped homodimer in the crystal and also in solution. Through a detailed mutational analysis of all interface residues, we identified several hydrophobic residues that are important for AHR dimerization and function. Our crystallographic visualization of AHR PAS-A dimerization leads us to propose a mode of heterodimerization with ARNT that is supported by both biochemical and cell-based data. Our studies also highlight the residues of other mammalian bHLH-PAS proteins that are likely involved in their homo- or heterodimerization. PMID:24001774

  6. The Drosophila dysfusion basic helix-loop-helix (bHLH)-PAS gene controls tracheal fusion and levels of the trachealess bHLH-PAS protein.

    PubMed

    Jiang, Lan; Crews, Stephen T

    2003-08-01

    The development of the mature insect trachea requires a complex series of cellular events, including tracheal cell specification, cell migration, tubule branching, and tubule fusion. Here we describe the identification of the Drosophila melanogaster dysfusion gene, which encodes a novel basic helix-loop-helix (bHLH)-PAS protein conserved between Caenorhabditis elegans, insects, and humans, and controls tracheal fusion events. The Dysfusion protein functions as a heterodimer with the Tango bHLH-PAS protein in vivo to form a putative DNA-binding complex. The dysfusion gene is expressed in a variety of embryonic cell types, including tracheal-fusion, leading-edge, foregut atrium cells, nervous system, hindgut, and anal pad cells. RNAi experiments indicate that dysfusion is required for dorsal branch, lateral trunk, and ganglionic branch fusion but not for fusion of the dorsal trunk. The escargot gene, which is also expressed in fusion cells and is required for tracheal fusion, precedes dysfusion expression. Analysis of escargot mutants indicates a complex pattern of dysfusion regulation, such that dysfusion expression is dependent on escargot in the dorsal and ganglionic branches but not the dorsal trunk. Early in tracheal development, the Trachealess bHLH-PAS protein is present at uniformly high levels in all tracheal cells, but since the levels of Dysfusion rise in wild-type fusion cells, the levels of Trachealess in fusion cells decline. The downregulation of Trachealess is dependent on dysfusion function. These results suggest the possibility that competitive interactions between basic helix-loop-helix-PAS proteins (Dysfusion, Trachealess, and possibly Similar) may be important for the proper development of the trachea. PMID:12897136

  7. Flavonoid-specific staining of Arabidopsis thaliana.

    PubMed

    Sheahan, J J; Rechnitz, G A

    1992-12-01

    Crop yields may be threatened by increases in UV-B radiation resulting from depletion of the ozone layer. In higher plants, the presence of flavonols provides a protective mechanism, and we report a novel staining procedure for the visualization of such protectants in plant tissue. It is shown that the proposed technique provides sensitive and specific fluorescence of flavonoids in chlorophyll-bleached tissue of Arabidopsis thaliana. PMID:1282347

  8. Laser Treatment of Port Wine Stains

    NASA Astrophysics Data System (ADS)

    Majaron, Boris; Nelson, J. Stuart

    Port wine stain (PWS), also called nevus flammeus, is a congenital, cutaneous vascular malformation involving post-capillary venules which produce a light pink to red to dark-red-violet discoloration of human skin [1]. PWS occurs in an estimated 3 children per 1000 live births, affecting males and females and all racial groups equally [2]. There appears to be no hereditary predilection for PWS within families. There are no known risk factors or ways to prevent PWS.

  9. Methods and compositions for chromosome-specific staining

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel

    2003-07-22

    Methods and compositions for chromosome-specific staining are provided. Compositions comprise heterogenous mixtures of labeled nucleic acid fragments having substantially complementary base sequences to unique sequence regions of the chromosomal DNA for which their associated staining reagent is specific. Methods include methods for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes.

  10. Faux pas detection and intentional action in Asperger Syndrome. A replication on a French sample.

    PubMed

    Zalla, Tiziana; Sav, Anca-Maria; Stopin, Astrid; Ahade, Sabrina; Leboyer, Marion

    2009-02-01

    In the present study, we investigated mind reading abilities in a group of adults with Asperger Syndrome (AS) by using the faux pas task, an advanced test of theory of mind (Baron-Cohen et al. (1999). Journal of Autism and Developmental Disorders, 29, 407-418). The faux pas is a particular case of a non-intentional action reflecting an involuntary socially inappropriate behavior. Here, individuals with AS over-detected faux pas stories, failed to provide correct justifications of the speaker's behavior and were unaware of the mistaken belief and of the resulting emotional impact, whereas they appeared to be responsive to social rule violations. We hypothesized that because of an impaired theory-of-mind, individuals with AS may develop compensatory cognitive strategies based on overlearned abstract knowledge about normative rules. PMID:18726150

  11. Space Data Systems Applications in the iPAS Pathfinder Laboratory

    NASA Technical Reports Server (NTRS)

    Rich, Tom

    2013-01-01

    The iPAS is an integrated hardware/software test and evaluation environment, in support of current and future spacecraft development The iPAS has two main elements. A common avionics, hardware, and software architecture that can be applied over various missions. A common testbed framework that supports integrated hardware/software testing for a variety of applications. The iPAS includes the following (non-flight qualified) components: Core Flight Software (from GSFC). Commercially available Proton and S950 Flight Computer boards. Power and propulsion systems based on representative flight hardware. A realistic flight deck based on the Multi-Purpose Crew Vehicle (MPCV), including realistic flight controls and displays. A Space Data System based on CCSDS protocols.

  12. Ultrastructural evidence of Tropheryma whippelii in PAS-negative granulomatous lymph nodes.

    PubMed

    Finzi, Giovanna; Franzi, Francesca; Sessa, Fausto; Mastaglio, Claudio; Capella, Carlo

    2007-01-01

    The presence of Tropheryma whippelii was demonstrated in the PAS-negative mesenteric granulomatous lymph nodes of a patient affected by Whipple disease. Ultrastructurally a few bacteria, enclosed by a membrane characteristic of Tropheryma whippelii, were found in the extracellular spaces and remnants of bacteria were found in the phagocytic vacuoles of macrophages. The scarce number of bacilli, probably due to the fact that the disease was at an initial phase, could explain the absence of PAS positivity. This case confirms the role of the electron microscopy in the diagnosis of Whipple disease, especially for extra-intestinal lesions and at the initial phase of the disease, when the characteristic PAS-positive macrophages can be absent. PMID:17613996

  13. PAS/LOV proteins: A proposed new class of plant blue light receptor.

    PubMed

    Ogura, Yasunobu; Tokutomi, Satoru; Wada, Masamitsu; Kiyosue, Tomohiro

    2008-11-01

    The light, oxygen or voltage (LOV) domain belongs to the Per-ARNT-Sim (PAS) superfamily of domains, and functions with the flavin chromophore as a module for sensing blue light in plants and fungi. The Arabidopsis thaliana PAS/LOV proteins (PLPs), of unknown function, possess an N-terminal PAS domain and a C-terminal LOV domain. Our recent analysis using yeast two-hybrid and Escherichia coli protein production systems reveals that the interactions of Arabidopsis PLPs with several proteins diminish under blue light illumination and that the PLP LOV domain may bind to a flavin chromophore. These results suggest that PLP functions as a blue light receptor. Homologs of PLP exist in rice, tomato and moss. The LOV domains of these PLP homologs form a distinct group in phylogenetic analysis. These facts suggest that PLP belongs to a new class of plant blue light receptor. PMID:19704421

  14. Newer applications of the histological stain prepared from Pterocarpus santalinus.

    PubMed

    Sen Gupta, P C; Mukherjee, A K

    1981-03-01

    A histological stain prepared from the heartwood of Pterocarpus santalinus Linn. has been found to be an excellent nuclear stain for various cells of animal and plant origin. As an elastic tissue stain, the results are comparable to standard elastic tissue stains. The striations of voluntary muscle fibers are well shown. The Nissl granules and fibers of cranial nerves in the pons are visualized. When counterstained with light green, it differentially stains muscle and fibrous tissue. The stain can be used as counterstain with certain histochemical procedures with satisfactory results. The preparation and use of this versatile stain are described. PMID:6166099

  15. Evaluation of lanthanide salts as alternative stains to uranyl acetate.

    PubMed

    Hosogi, Naoki; Nishioka, Hideo; Nakakoshi, Masamichi

    2015-12-01

    Uranyl acetate (UAc) has been generally used not only as a superb staining reagent for ultrathin sections of plastic-embedded biological materials, but also as high-contrast negative stains for biological macromolecules such as particles of protein or virus. However, the use and purchase of radioactive UAc have been restricted. In this study, we determine the performance of ytterbium triacetate, lutetium triacetate, samarium triacetate and gadolinium triacetate as new staining reagents for biological electron microscopy. We observed chemically fixed spinach (Spinacia oleracea) leaves stained with these reagents. Ultrathin sections were stained with these reagents. Some of them were counterstained with lead citrate. The transmission electron microscopy contrast of spinach organelles was evaluated in sections exposed to the conventional stain and new stains. We show acetate salts of samarium, gadolinium, ytterbium and lutetium could be excellent substitutes for UAc for thin section staining and for negative staining. In addition, each reagent showed appreciable negative-staining effects. PMID:26374081

  16. Pituitary Adenoma With Paraganglioma/Pheochromocytoma (3PAs) and Succinate Dehydrogenase Defects in Humans and Mice

    PubMed Central

    Xekouki, Paraskevi; Szarek, Eva; Bullova, Petra; Giubellino, Alessio; Quezado, Martha; Mastroyannis, Spyridon A.; Mastorakos, Panagiotis; Wassif, Christopher A.; Raygada, Margarita; Rentia, Nadia; Dye, Louis; Cougnoux, Antony; Koziol, Deloris; Sierra, Maria de La Luz; Lyssikatos, Charalampos; Belyavskaya, Elena; Malchoff, Carl; Moline, Jessica; Eng, Charis; Maher, Louis James; Pacak, Karel; Lodish, Maya

    2015-01-01

    Context: Germline mutations in genes coding succinate dehydrogenase (SDH) subunits A, B, C, and D have been identified in familial paragangliomas (PGLs)/pheochromocytomas (PHEOs) and other tumors. We described a GH-secreting pituitary adenoma (PA) caused by SDHD mutation in a patient with familial PGLs. Additional patients with PAs and SDHx defects have since been reported. Design: We studied 168 patients with unselected sporadic PA and with the association of PAs, PGLs, and/or pheochromocytomas, a condition we named the 3P association (3PAs) for SDHx germline mutations. We also studied the pituitary gland and hormonal profile of Sdhb+/− mice and their wild-type littermates at different ages. Results: No SDHx mutations were detected among sporadic PA, whereas three of four familial cases were positive for a mutation (75%). Most of the SDHx-deficient PAs were either prolactinomas or somatotropinomas. Pituitaries of Sdhb+/− mice older than 12 months had an increased number mainly of prolactin-secreting cells and several ultrastructural abnormalities such as intranuclear inclusions, altered chromatin nuclear pattern, and abnormal mitochondria. Igf-1 levels of mutant mice tended to be higher across age groups, whereas Prl and Gh levels varied according to age and sex. Conclusion: The present study confirms the existence of a new association that we termed 3PAs. It is due mostly to germline SDHx defects, although sporadic cases of 3PAs without SDHx defects also exist. Using Sdhb+/− mice, we provide evidence that pituitary hyperplasia in SDHx-deficient cells may be the initial abnormality in the cascade of events leading to PA formation. PMID:25695889

  17. J-PAS: The Javalambre-Physics of the Accelerating Universe Astrophysical Survey

    NASA Astrophysics Data System (ADS)

    Dupke, Renato A.; Benitez, Narciso; Moles, Mariano; Sodre, Laerte; Irwin, Jimmy; J-PAS Collaboration

    2016-01-01

    The Javalambre-Physics of the Accelerating Universe Astrophysical Survey (J-PAS) is a narrow band, very wide field Cosmological Survey to be carried out from the Javalambre Astrophysical Observatory in Spain with a dedicated 2.5m telescope and a 4.7deg^2 camera with 1.2Gpix. Starting in 2016, J-PAS will observe 8600 deg^2 of the Northern Sky and measure 0.003(1+z) precision photometric redshifts for nearly 1E08 LRG and ELG galaxies plus several million QSOs, sampling an effective volume of ~14 Gpc^3 up to z = 1.3. J-PAS will also detect and measure the mass of more than a hundred thousand galaxy clusters, setting constrains on Dark Energy which rival those obtained from BAO measurements.The key to the J-PAS potential is its innovative approach the combination of 54 145°A filters, placed 100°A apart, and a multi-degree field of view (FOV) which makes it a powerful "redshift machine", with the survey speed of a 4000 multiplexing low resolution spectrograph, but many times cheaper and much faster to build. Moreover, since the J-PAS camera is equivalent to a very large, 4.7deg^2 "IFU", it will produce a time-resolved, 3D image of the Northern Sky with a very wide range of Astrophysical applications in Galaxy Evolution, the nearby Universe and the study of resolved stellar populations. J-PAS will have a lasting legacy value in many areas of Astrophysics, serving as a fundamental dataset for future Cosmological projects.Here, we present the overall description, status and scientific potential of the survey.

  18. J-PAS: The Javalambre-Physics of the Accelerating Universe Astrophysical Survey

    NASA Astrophysics Data System (ADS)

    Dupke, Renato a.; Benitez, Narciso; Moles, Mariano; Sodre, Laerte; J-PAS Collaboration

    2015-08-01

    The Javalambre-Physics of the Accelerating Universe Astrophysical Survey (J-PAS) is a narrow band, very wide field Cosmological Survey to be carried out from the Javalambre Astrophysical Observatory in Spain with a dedicated 2.5m telescope and a 4.7deg^2 camera with 1.2Gpix. Starting in 2016, J-PAS will observe 8600 deg^2 of the Northern Sky and measure 0.003(1+z) precision photometric redshifts for nearly 1E08 LRG and ELG galaxies plus several million QSOs, sampling an effective volume of ~14 Gpc^3 up to z = 1.3. J-PAS will also detect and measure the mass of more than a hundred thousand galaxy clusters, setting constrains on Dark Energy which rival those obtained from BAO measurements.The key to the J-PAS potential is its innovative approach the combination of 54 145°A filters, placed 100°A apart, and a multi-degree field of view (FOV) which makes it a powerful “redshift machine”, with the survey speed of a 4000 multiplexing low resolution spectrograph, but many times cheaper and much faster to build. Moreover, since the J-PAS camera is equivalent to a very large, 4.7deg^2 “IFU”, it will produce a time-resolved, 3D image of the Northern Sky with a very wide range of Astrophysical applications in Galaxy Evolution, the nearby Universe and the study of resolved stellar populations. J-PAS will have a lasting legacy value in many areas of Astrophysics, serving as a fundamental dataset for future Cosmological projects.Here, we present the overall description, status and scientific potential of the survey.

  19. Digital radiography using amorphous selenium: photoconductively activated switch (PAS) readout system.

    PubMed

    Reznik, Nikita; Komljenovic, Philip T; Germann, Stephen; Rowlands, John A

    2008-03-01

    A new amorphous selenium (a-Se) digital radiography detector is introduced. The proposed detector generates a charge image in the a-Se layer in a conventional manner, which is stored on electrode pixels at the surface of the a-Se layer. A novel method, called photoconductively activated switch (PAS), is used to read out the latent x-ray charge image. The PAS readout method uses lateral photoconduction at the a-Se surface which is a revolutionary modification of the bulk photoinduced discharge (PID) methods. The PAS method addresses and eliminates the fundamental weaknesses of the PID methods--long readout times and high readout noise--while maintaining the structural simplicity and high resolution for which PID optical readout systems are noted. The photoconduction properties of the a-Se surface were investigated and the geometrical design for the electrode pixels for a PAS radiography system was determined. This design was implemented in a single pixel PAS evaluation system. The results show that the PAS x-ray induced output charge signal was reproducible and depended linearly on the x-ray exposure in the diagnostic exposure range. Furthermore, the readout was reasonably rapid (10 ms for pixel discharge). The proposed detector allows readout of half a pixel row at a time (odd pixels followed by even pixels), thus permitting the readout of a complete image in 30 s for a 40 cm x 40 cm detector with the potential of reducing that time by using greater readout light intensity. This demonstrates that a-Se based x-ray detectors using photoconductively activated switches could form a basis for a practical integrated digital radiography system. PMID:18404939

  20. Digital radiography using amorphous selenium: Photoconductively activated switch (PAS) readout system

    SciTech Connect

    Reznik, Nikita; Komljenovic, Philip T.; Germann, Stephen; Rowlands, John A.

    2008-03-15

    A new amorphous selenium (a-Se) digital radiography detector is introduced. The proposed detector generates a charge image in the a-Se layer in a conventional manner, which is stored on electrode pixels at the surface of the a-Se layer. A novel method, called photoconductively activated switch (PAS), is used to read out the latent x-ray charge image. The PAS readout method uses lateral photoconduction at the a-Se surface which is a revolutionary modification of the bulk photoinduced discharge (PID) methods. The PAS method addresses and eliminates the fundamental weaknesses of the PID methods--long readout times and high readout noise--while maintaining the structural simplicity and high resolution for which PID optical readout systems are noted. The photoconduction properties of the a-Se surface were investigated and the geometrical design for the electrode pixels for a PAS radiography system was determined. This design was implemented in a single pixel PAS evaluation system. The results show that the PAS x-ray induced output charge signal was reproducible and depended linearly on the x-ray exposure in the diagnostic exposure range. Furthermore, the readout was reasonably rapid (10 ms for pixel discharge). The proposed detector allows readout of half a pixel row at a time (odd pixels followed by even pixels), thus permitting the readout of a complete image in 30 s for a 40 cmx40 cm detector with the potential of reducing that time by using greater readout light intensity. This demonstrates that a-Se based x-ray detectors using photoconductively activated switches could form a basis for a practical integrated digital radiography system.

  1. Neuropathology in respiratory-related motoneurons in young Pompe (Gaa(-/-)) mice.

    PubMed

    Turner, Sara M F; Hoyt, Aaron K; ElMallah, Mai K; Falk, Darin J; Byrne, Barry J; Fuller, David D

    2016-06-15

    Respiratory and/or lingual dysfunction are among the first motor symptoms in Pompe disease, a disorder resulting from absence or dysfunction of the lysosomal enzyme acid α-glucosidase (GAA). Here, we histologically evaluated the medulla, cervical and thoracic spinal cords in 6 weeks old asymptomatic Pompe (Gaa(-/-)) mice to determine if neuropathology in respiratory motor regions has an early onset. Periodic acid-Schiff (PAS) staining indicated glycogen accumulation was exclusively occurring in Gaa(-/-) hypoglossal, mid-cervical and upper thoracic motoneurons. Markers of DNA damage (Tunel) and ongoing apoptosis (Cleaved Caspase 3) did not co-localize with PAS staining, but were prominent in a medullary region which included the nucleus tractus solitarius, and also in the thoracic spinal dorsal horn. We conclude that respiratory-related motoneurons are particularly susceptible to GAA deficiency and that neuronal glycogen accumulation and neurodegeneration may occur independently in early stage disease. The data support early therapeutic intervention in Pompe disease. PMID:26921786

  2. Bleaching of fluorosis stains using sodium hypochlorite

    PubMed Central

    Penumatsa, Narendra Varma; Sharanesha, Rajashekhara Bhari

    2015-01-01

    Fluorosis staining is commonly considered an esthetic problem because of the psychological impact of unesthetic maxillary anterior teeth. Numerous treatment approaches have been proposed, ranging from bleaching to enamel reduction to restorative techniques. Bleaching of hypomineralized enamel lesions, using 5% sodium hypochlorite, has been useful clinically. The technique described, in this case, appears to have advantages over other methods for improving the appearance of fluorotic lesions. It is simple, low cost, noninvasive, so the enamel keeps its structure, relatively rapid, and safe; it requires no special materials, and it can be used with safety on young permanent teeth. PMID:26538964

  3. False-positive Gram-stained smears.

    PubMed

    Hoke, C H; Batt, J M; Mirrett, S; Cox, R L; Reller, L B

    1979-02-01

    The rate per 1,000 smears showing nonviable Gram-negative bacilli (false-positive smears) increased from a baseline of 10.8 to 38.5 following purchase of new culture-collection devices; the rate decreased to 8.0 following replacement of contaminated culture sets. False-positive reports led to changes in therapy for five patients. In addition to being sterile, commercial culture-collection devices should be certified by the manufacturer as being free of stainable microorganisms or as unsuitable for preparation of Gram-stained smears. PMID:83398

  4. Histological Stains: A Literature Review and Case Study

    PubMed Central

    Alturkistani, Hani A; Tashkandi, Faris M; Mohammedsaleh, Zuhair M

    2016-01-01

    The history of histology indicates that there have been significant changes in the techniques used for histological staining through chemical, molecular biology assays and immunological techniques, collectively referred to as histochemistry. Early histologists used the readily available chemicals to prepare tissues for microscopic studies; these laboratory chemicals were potassium dichromate, alcohol and the mercuric chloride to harden cellular tissues. Staining techniques used were carmine, silver nitrate, Giemsa, Trichrome Stains, Gram Stain and Hematoxylin among others. The purpose of this research was to assess past and current literature reviews, as well as case studies, with the aim of informing ways in which histological stains have been improved in the modern age. Results from the literature review has indicated that there has been an improvement in histopathology and histotechnology in stains used. There has been a rising need for efficient, accurate and less complex staining procedures. Many stain procedures are still in use today, and many others have been replaced with new immunostaining, molecular, non-culture and other advanced staining techniques. Some staining methods have been abandoned because the chemicals required have been medically proven to be toxic. The case studies indicated that in modern histology a combination of different stain techniques are used to enhance the effectiveness of the staining process. Currently, improved histological stains, have been modified and combined with other stains to improve their effectiveness. PMID:26493433

  5. Psychometric Properties and Norms of the German ABC-Community and PAS-ADD Checklist

    ERIC Educational Resources Information Center

    Zeilinger, Elisabeth L.; Weber, Germain; Haveman, Meindert J.

    2011-01-01

    Aim: The aim of the present study was to standardize and generate psychometric evidence of the German language versions of two well-established English language mental health instruments: the "Aberrant Behavior Checklist-Community" (ABC-C) and the "Psychiatric Assessment Schedule for Adults with Developmental Disabilities" (PAS-ADD) Checklist. New…

  6. Crystal structure of the PAS domain of the hEAG potassium channel.

    PubMed

    Tang, Xue; Shao, Juan; Qin, Xiaohong

    2016-08-01

    KCNH voltage-gated potassium channels play critical roles in regulating cellular functions. The channel is composed of four subunits, each of which contains six transmembrane helices forming the central pore. The cytoplasmic parts of the subunits present a Per-Arnt-Sim (PAS) domain at the N-terminus and a cyclic nucleotide-binding homology domain at the C-terminus. PAS domains are conserved from prokaryotes to eukaryotes and are involved in sensing signals and cellular responses. To better understand the functional roles of PAS domains in KCNH channels, the structure of this domain from the human ether-à-go-go channel (hEAG channel) was determined. By comparing it with the structures of the Homo sapiens EAG-related gene (hERG) channel and the Drosophila EAG-like K(+) (dELK) channel and analyzing the structural features of the hEAG channel, it was identified that a hydrophobic patch on the β-sheet may mediate interaction between the PAS domain and other regions of the channel to regulate its functions. PMID:27487920

  7. Crystal structure of the PAS domain of the hEAG potassium channel

    PubMed Central

    Tang, Xue; Shao, Juan; Qin, Xiaohong

    2016-01-01

    KCNH voltage-gated potassium channels play critical roles in regulating cellular functions. The channel is composed of four subunits, each of which contains six transmembrane helices forming the central pore. The cytoplasmic parts of the subunits present a Per–Arnt–Sim (PAS) domain at the N-terminus and a cyclic nucleotide-binding homology domain at the C-terminus. PAS domains are conserved from prokaryotes to eukaryotes and are involved in sensing signals and cellular responses. To better understand the functional roles of PAS domains in KCNH channels, the structure of this domain from the human ether-à-go-go channel (hEAG channel) was determined. By comparing it with the structures of the Homo sapiens EAG-related gene (hERG) channel and the Drosophila EAG-like K+ (dELK) channel and analyzing the structural features of the hEAG channel, it was identified that a hydrophobic patch on the β-sheet may mediate interaction between the PAS domain and other regions of the channel to regulate its functions. PMID:27487920

  8. 32 CFR 806b.11 - When to give Privacy Act Statements (PAS).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 6 2010-07-01 2010-07-01 false When to give Privacy Act Statements (PAS). 806b... ADMINISTRATION PRIVACY ACT PROGRAM Collecting Personal Information § 806b.11 When to give Privacy Act Statements... information. Give a copy of the Privacy Act Statement if asked. Do not ask the person to sign the Privacy...

  9. 32 CFR 806b.11 - When to give Privacy Act Statements (PAS).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 6 2011-07-01 2011-07-01 false When to give Privacy Act Statements (PAS). 806b... ADMINISTRATION PRIVACY ACT PROGRAM Collecting Personal Information § 806b.11 When to give Privacy Act Statements... information. Give a copy of the Privacy Act Statement if asked. Do not ask the person to sign the Privacy...

  10. Searching for planetary nebulae at the Galactic halo via J-PAS

    NASA Astrophysics Data System (ADS)

    Gonçalves, Denise R.; Aparício-Villegas, T.; Akras, S.; Cortesi, A.; Borges-Fernandes, M.; Daflon, S.; Pereira, C. B.; Lorenz-Martins, S.; Marcolino, W.; Kanaan, A.; Viironen, K.; de Oliveira, C. Mendes; Molino, A.; Ederoclite, A.

    2016-08-01

    The Javalambre-Physics of the Accelerating Universe Astrophysical Survey (J-PAS) is a narrow-band imaging, very wide field cosmological survey. It will last 5 years and will observe 8500 sq. deg. of the sky. There will be 54 contiguous narrow-band filters of 145Å FWHM, from 3,500 to 10,000Å. Two broad-band filters will be added at the extremes, UV and IR, plus the 3-g, r, and i- SDSS filters. Thus, J-PAS can be an important tool to search for new planetary nebulae (PNe) at the halo, increasing their numbers, because only 14 of them have been convincingly identified in the literature. Halo PNe are able to reveal precious information for the study of stellar evolution and the early chemical conditions of the Galaxy. The characteristic low continuum and intense emission lines of PNe make them good objects to be searched by J-PAS. Though covering a significantly smaller sky area, data from the ALHAMBRA survey were used to test our J-PAS strategy to search for PNe. Our first results are shown in this contribution.

  11. Test report for PAS-1 cask certification for shipping payload B

    SciTech Connect

    MERCADO, J.E.

    1998-10-13

    This test report documents the successful inspection and testing to certify two NuPac PAS-1 casks in accordance with US Department of Energy Certificate of Compliance (CoC) USA/9184/B(U). The primary and secondary containment vessels of each cask met the acceptance criteria defined in the CoC and the test plan.

  12. J-PAS: The Javalambre-Physics of the Accelerated Universe Astrophysical Survey

    NASA Astrophysics Data System (ADS)

    Benítez, N.; Dupke, R.; Moles, M.; Sodré, L.; Cenarro, A. J.; Marín Franch, A.; Taylor, K.; Cristóbal, D.; Fernández-Soto, A.; Mendes de Oliveira, C.; Cepa-Nogué, J.; Abramo, L. R.; Alcaniz, J. S.; Overzier, R.; Hernández-Monteagudo, C.; Alfaro, E. J.; Kanaan, A.; Carvano, M.; Reis, R. R. R.; J-PAS Collaboration

    2015-05-01

    J-PAS is a Spanish-Brazilian 8500 deg^2 Cosmological Survey which will be carried out from the Javalambre Observatory with a purpose-built, dedicated 2.5 m telescope and a 4.7 deg^2 camera with 1.2 Gpix. Starting in 2015, J-PAS will use 59 filters to measure high precision 0.003(1+z) photometric redshifts for 90M galaxies plus several million QSOs, about 50 times more than the largest current spectroscopic survey, sampling an effective volume of ˜ 14 Gpc^3 up to z=1.3. J-PAS will not only be first radial BAO experiment to reach Stage IV; it will also detect and measure the mass of 7× 10^5 galaxy clusters and groups, setting constrains on Dark Energy which rival those obtained from BAO measurements. The combination of a set of 145 Å NB filters, placed 100 Å apart, and a multi-degree field of view is a powerful ``redshift machine'', equivalent to a 4000 multiplexing spectrograph, but many times cheaper to build. The J-PAS camera is equivalent to a very large, 4.7 deg^2 ``IFU'', which will produce a time-resolved, 3D image of the Northern Sky with a very wide range of scientific applications in Galaxy Evolution, Stellar Physics and the Solar System.

  13. Peer Relations and the Understanding of Faux Pas: Longitudinal Evidence for Bidirectional Associations

    ERIC Educational Resources Information Center

    Banerjee, Robin; Watling, Dawn; Caputi, Marcella

    2011-01-01

    Research connecting children's understanding of mental states to their peer relations at school remains scarce. Previous work by the authors demonstrated that children's understanding of mental states in the context of a faux pas--a social blunder involving unintentional insult--is associated with concurrent peer rejection. The present report…

  14. Exploratory studies of extended storage of apheresis platelets in a platelet additive solution (PAS).

    PubMed

    Slichter, Sherrill J; Corson, Jill; Jones, Mary Kay; Christoffel, Todd; Pellham, Esther; Bailey, S Lawrence; Bolgiano, Doug

    2014-01-01

    To evaluate the poststorage viability of apheresis platelets stored for up to 18 days in 80% platelet additive solution (PAS)/20% plasma, 117 healthy subjects donated platelets using the Haemonetics MCS+, COBE Spectra (Spectra), or Trima Accel (Trima) systems. Control platelets from the same subjects were compared with their stored test PAS platelets by radiolabeling their stored and control platelets with either (51)chromium or (111)indium. Trima platelets met Food and Drug Administration poststorage platelet viability criteria for only 7 days vs almost 13 days for Haemonetics platelets; ie, platelet recoveries after these storage times averaged 44 ± 3% vs 49 ± 3% and survivals were 5.4 ± 0.3 vs 4.6 ± 0.3 days, respectively. The differences in storage duration are likely related to both the collection system and the storage bag. The Spectra and Trima platelets were hyperconcentrated during collection, and PAS was added, whereas the Haemonetics platelets were elutriated with PAS, which may have resulted in less collection injury. When Spectra and Trima platelets were stored in Haemonetics' bags, poststorage viability was significantly improved. Platelet viability is better maintained in vitro than in vivo, allowing substantial increases in platelet storage times. However, implementation will require resolution of potential bacterial overgrowth during storage. PMID:24258816

  15. Stain removal from a pigmented silicone maxillofacial elastomer.

    PubMed

    Yu, R; Koran, A; Craig, R G; Raptis, C N

    1982-08-01

    The removal of environmental stains from a pigmented maxillofacial elastomer was carried out by solvent extraction under network swelling. Silastic 44210 was pigmented with 11 maxillofacial pigments prior to staining. Samples were stained with lipstick, methylene blue, and disclosing solution. These stains were then removed by solvent extraction with 1,1,1-trichloroethane. Color parameter measurements both before and after staining and after solvent extraction demonstrated the effectiveness of removing these stains by solvent extraction while causing little or no change in the color of the pigmented samples. PMID:6955345

  16. Treatment of port-wine stains: analysis

    SciTech Connect

    van Gemert, M.J.; Welch, A.J.

    1987-08-01

    Port-wine stains (PWS) are bluish red skin stains that are caused by enlarged, ectatic blood vessels in the dermis. Laser treatment of PWS is analyzed from computation of the spatial distribution of heat production by direct absorption of the laser light and subsequent heat conduction. The absorption and scattering caused by oxyhemoglobin, epidermis, and dermis as a function of wavelength are utilized in this analysis. Ideal treatment is defined as coagulating the ectatic blood vessels without irreversible damage to the epidermis and dermis. The analysis shows that a millisecond pulsed, yellow dye laser at 577 nm (one of the large absorption bands in blood) is the laser of choice to treat PWS, offering as close to the ''ideal treatment'' as possible. The blue-green argon laser, which is currently the most frequently used laser for this purpose, is strongly recommended with irradiation times in milliseconds. Other lasers that are in clinical use, such as the red ruby and near-infrared Nd-YAG lasers, can provide selective treatment only when the epidermis is cooled concurrently. The CO/sub 2/ laser, on the other hand, can coagulate the blood vessels only through heat conduction from the hot epidermis; hence, it has neither the treatment selectivity nor any other physical option to force this selectivity.

  17. Ki-67 Membranous Staining: Biologically Relevant or an Artifact of Multiplexed Immunofluorescent Staining.

    PubMed

    Wang, Dan; Pang, Zhengyu; Clarke, Gina M; Nofech-Mozes, Sharon; Liu, Kela; Cheung, Alison M Y; Filkins, Robert J; Yaffe, Martin J

    2016-07-01

    In the process of developing a multiplex of 8 common breast cancer biomarkers (Her2/neu, estrogen receptor, progesterone receptor, Ki-67, aldehyde dehydrogenase-1, NaK-ATPase, cytokeratin 8/18, and myosin smooth muscle) on a single formalin-fixed paraffin-embedded slide using a sequential staining, imaging, and dye bleaching technology developed by General Electric Company, membranous Ki-67 staining was observed and colocalized with Her2/neu staining. Using immunohistochemistry as gold standards, we discovered that membranous Ki-67 was an artifact caused by the binding of cyanine 5-conjugated rabbit polyclonal Ki-67 antibody to a secondary cyanine 3-conjugated donkey anti-rabbit antibody which was previously applied and bound to rabbit Her2/neu antibody in our multiplexing experiment. After blocking with rabbit serum, a successful protocol for 8 biomarker multiplexing without cross-reactivity of antibodies from the same species was developed. PMID:26258752

  18. Staining Protocols for Human Pancreatic Islets

    PubMed Central

    Campbell-Thompson, Martha L.; Heiple, Tiffany; Montgomery, Emily; Zhang, Li; Schneider, Lynda

    2012-01-01

    Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and beta mass ranges from 500 to 1500 mg 1-3. With this known heterogeneity, a standard processing and staining procedure was developed so that pancreatic regions were clearly defined and islets characterized using rigorous histopathology and immunolocalization examinations. Standardized procedures for processing human pancreas recovered from organ donors are described in part 1 of this series. The pancreas is processed into 3 main regions (head, body, tail) followed by transverse sections. Transverse sections from the pancreas head are further divided, as indicated based on size, and numbered alphabetically to denote subsections. This standardization allows for a complete cross sectional analysis of the head region including the uncinate region which contains islets composed primarily of pancreatic polypeptide cells to the tail region. The current report comprises part 2 of this series and describes the procedures used for serial sectioning and histopathological characterization of the pancreatic paraffin sections with an emphasis on islet endocrine cells, replication, and T-cell infiltrates. Pathology of pancreatic sections is intended to characterize both exocrine, ductular, and endocrine components. The exocrine compartment is evaluated for the presence of pancreatitis (active or chronic), atrophy, fibrosis, and fat, as well as the duct system, particularly in relationship to the presence of pancreatic intraductal neoplasia4. Islets are evaluated for morphology, size, and density, endocrine cells, inflammation, fibrosis, amyloid, and the presence of replicating or apoptotic cells using H&E and IHC stains. The final component described in part 2 is the provision of the stained slides as digitized whole slide images. The digitized slides are organized by case and pancreas region in an online pathology database

  19. Does Faux Pas Detection in Adult Autism Reflect Differences in Social Cognition or Decision-Making Abilities?

    ERIC Educational Resources Information Center

    Thiébaut, Flora I.; White, Sarah J.; Walsh, Annabel; Klargaard, Solja K.; Wu, Hsuan-Chen; Rees, Geraint; Burgess, Paul W.

    2016-01-01

    43 typically-developed adults and 35 adults with ASD performed a cartoon faux pas test. Adults with ASD apparently over-detected faux pas despite good comprehension abilities, and were generally slower at responding. Signal detection analysis demonstrated that the ASD participants had significantly greater difficulty detecting whether a cartoon…

  20. Port wine stain on a child's face (image)

    MedlinePlus

    Port wine stains are always present at birth. In an infant, they are flat, pink, vascular lesions. Common locations ... may be present anywhere on the body. Port wine stains may appear in association with other syndromes.

  1. Several staining techniques to enhance the visibility of Acanthamoeba cysts.

    PubMed

    El-Sayed, Nagwa Mostafa; Hikal, Wafaa Mohamed

    2015-03-01

    Acanthamoeba is one of the most common free-living amoebae. It is widespread in the environment and can infect humans causing keratitis. Delayed diagnosis or misdiagnosis leads to extensive corneal inflammation and profound visual loss. Therefore, accurate and rapid diagnosis of Acanthamoeba keratitis is essential for successful treatment and good prognosis. This study was designed to use different staining techniques to facilitate the identification of Acanthamoeba cysts. Acanthamoeba cysts were isolated by cultivation of either corneal scraping specimens or tap water samples onto non-nutrient agar plates seeded with Escherichia coli. Subcultures were done from positive cultures until unique cysts were isolated. Acanthamoeba cysts were stained temporarily using iodine, eosin, methylene blue, and calcofluor white (CFW) stains and as permanent slides after processing for mounting using modified trichrome, Gimenez and Giemsa staining. These stains were compared on the basis of staining quality including clarity of morphological details, differentiation between cytoplasm and nuclei, color and contrast, and also other characteristics of the staining techniques, including ease of handling, time taken for the procedure, and cost effectiveness. The cysts of Acanthamoeba were recognized in the form of double-walled cysts: the outer wall (ectocyst) that was being differentiated from the variably stained surrounding background and the inner wall (endocyst) that was sometimes stellated, polygonal, round, or oval and visualized as separate from the spherical, sometimes irregular, outline of the ectocyst. Regarding the temporary stains, it was found that they were efficient for visualizing the morphological details of Acanthamoeba cysts. In CFW staining, Acanthamoeba cysts appeared as bluish-white or turquoise oval halos although the internal detail was not evident. On the other hand, the results of permanent-stained slides showed the most consistent stain for identification of

  2. Ultrafast tissue staining with chemical tags

    PubMed Central

    Kohl, Johannes; Ng, Julian; Cachero, Sebastian; Ciabatti, Ernesto; Dolan, Michael-John; Sutcliffe, Ben; Tozer, Adam; Ruehle, Sabine; Krueger, Daniel; Frechter, Shahar; Branco, Tiago; Tripodi, Marco; Jefferis, Gregory S. X. E.

    2014-01-01

    Genetically encoded fluorescent proteins and immunostaining are widely used to detect cellular and subcellular structures in fixed biological samples. However, for thick or whole-mount tissue, each approach suffers from limitations, including limited spectral flexibility and lower signal or slow speed, poor penetration, and high background labeling, respectively. We have overcome these limitations by using transgenically expressed chemical tags for rapid, even, high-signal and low-background labeling of thick biological tissues. We first construct a platform of widely applicable transgenic Drosophila reporter lines, demonstrating that chemical labeling can accelerate staining of whole-mount fly brains by a factor of 100. Using viral vectors to deliver chemical tags into the mouse brain, we then demonstrate that this labeling strategy works well in mice. Thus this tag-based approach drastically improves the speed and specificity of labeling genetically marked cells in intact and/or thick biological samples. PMID:25157152

  3. Does Faux Pas Detection in Adult Autism Reflect Differences in Social Cognition or Decision-Making Abilities?

    PubMed

    Thiébaut, Flora I; White, Sarah J; Walsh, Annabel; Klargaard, Solja K; Wu, Hsuan-Chen; Rees, Geraint; Burgess, Paul W

    2016-01-01

    43 typically-developed adults and 35 adults with ASD performed a cartoon faux pas test. Adults with ASD apparently over-detected faux pas despite good comprehension abilities, and were generally slower at responding. Signal detection analysis demonstrated that the ASD participants had significantly greater difficulty detecting whether a cartoon depicted a faux pas and showed a liberal response bias. Test item analysis demonstrated that the ASD group were not in agreement with a reference control group (n = 69) about which non-faux pas items were most difficult. These results suggest that the participants with ASD had a primary problem with faux pas detection, but that there is another factor at work, possibly compensatory, that relates to their choice of a liberal response criterion. PMID:26276266

  4. Investigation of a modified gallocyanin chrome alum staining technique in cytology compared to thionine and haematoxylin as nuclear stains.

    PubMed

    Schulte, E

    1988-01-01

    The present paper describes the staining characteristics of a modified Gallocyanin-chrome alum stain as compared to the original gallocyanin stain. Thionine, haematoxylin and the Feulgen reaction were used as controls. Tissue imprints of rabbit liver and spleen and smears of human venous blood were stained and controlled microscopically. Nuclear extinction was measured with the image analysis system IBAS 2000. Both GCA variants were examined by spectrophotometry and thin layer chromatography. The most striking difference between the GCA variants is the short staining time required for the modified stain (4 min) as compared to the original method (24 h). Both stains are stoichiometric for nucleic acids; the staining pattern, hue and intensity of nuclear colour and spectrophotometric and chromatographic data were absolutely consistent for both GCA-stains. These results and preliminary data from the analysis of the structure of the dye molecules seem to indicate that the molecular structure of the modified GCA is not changed by treatment with concentrated sulphuric acid. Differences in the staining kinetics might be due to differences in solubility. As nuclear chromatin texture after GCA staining is well appropriate for computerized image analysis the modified GCA-stain can be recommended as a simple and reproducible nuclear stain for automated feature extraction in cytology. PMID:2471227

  5. Cigarette staining and cleaning of a maxillofacial silicone

    SciTech Connect

    Yu, R.; Koran, A.; Raptis, C.N.; Craig, R.G.

    1983-07-01

    In this study, a maxillofacial silicone elastomer was stained with cigarette smoke. The stain was then removed by solvent extraction using 1,1,1-trichloroethane. The cigarette smoke produced large color changes in the elastomer as measured from spectrophotometric reflectance curves. The solvent was totally effective in removing the cigarette stain without changing the color of the silicone base.

  6. Searching for planetary nebulae at the Galactic halo via J-PAS and J-PLUS

    NASA Astrophysics Data System (ADS)

    Goncalves, Denise R.; Aparício-Villegas, Teresa; Akras, Stavros; Borges Fernandes, Marcelo; J-PAS Collaboration

    2015-08-01

    The Javalambre-Physics of the Accelerating Universe Astrophysical Survey (J-PAS) is a narrow-band imaging, very wide field cosmological survey to be carried out from a dedicated 2.5m telescope and a 4.7 sq.deg camera with 1.2Gpix. It will last 5 years and will observe 8500 sq.deg of Northern sky to a 5-σ magnitude depth for point sources, equivalent to i ~23.3 over an aperture of 2 arcsec2. The J-PAS filter system consists of 54 contiguous narrow band filters of 145-Å FWHM, from 3,500 to 10,000Å. Two broad-band filters will be added at the extremes, UV and IR, plus 3 SDSS g, r, and i filters. The Javalambre Photometric Local Universe Survye (J-PLUS) will be an auxiliary survey ofJ-PAS (mainly for calibration) with a dedicated 0.80m telescope. J-PLUS comprises 12 filters, including g, r, i and z SDSS ones. Though about 2,500 planetary nebulae (PNe, confirmed spectroscopically) are known in the Galaxy, only 14 objects have been convincingly identified as halo PNe. They were classified as such from their location, kinematics and chemistry. Halo PNe are able to reveal precious information for the study of low- and intermediate-mass star evolution and the early chemical conditions of the Galaxy. The characteristic low continuum and intense line emissions of PNe make them good objects to be searched by J-PAS, and even by J-PLUS. For instance, the halo PNe BoBn 1, DdDm 1 and PS 1, located somewhere between 11 and 24 kpc from the Sun, have B magnitudes of 16, 14 and 13.4, respectively. Such values are easily encompassed by J-PAS/J-PLUS, given the typical limit magnitude of the survey. Though covering a significantly smaller sky area, data from the ALHAMBRA survey were used to test our J-PAS/J-PLUS strategy to search for PNe. Our first results will be shown in this poster.

  7. Red food coloring stain: new, safer procedures for staining nematodes in roots and egg masses on root surfaces.

    PubMed

    Thies, Judy A; Merrill, Sharon B; Corley, E Luther

    2002-06-01

    Acid fuchsin and phloxine B are commonly used to stain plant-parasitic nematodes in roots and egg masses on root surfaces, respectively. Both stains can be harmful to both the user and the environment and require costly waste disposal procedures. We developed safer methods to replace both stains using McCormick Schilling red food color. Eggs, juveniles, and adults of Meloidogyne incognita stained in roots with red food color were equally as visible as those stained with acid fuchsin. Egg masses stained with red food color appeared as bright-red spheres on the root surfaces and were highly visible even without magnification. Replacement of acid fuchsin and phloxine B with red food color for staining nematodes is safer for the user and the environment, and eliminates costly waste disposal of used stain solutions. PMID:19265929

  8. Efficacy of rapid, economical, acetic acid, Papanicolaou stain in cervical smears as an alternative to conventional Papanicolaou stain

    PubMed Central

    Izhar, Shabnam; Kaur, Rupinder; Masih, Kanwal

    2014-01-01

    Background: Papanicolaou (Pap) stain has been used over the years for cervical cytology screening. However; it utilizes a considerable amount of alcohol which is expensive and difficult to procure. In one of the modifications, ethyl alcohol is replaced by 1% acetic acid and is termed as rapid, economical, acetic acid Papanicolaou (REAP) stain. It is cost effective, easily available and provides a suitable and rapid staining alternative. Aim: This study was undertaken to assess the efficacy of REAP stain as an alternative method to conventional Pap stain. Materials and Methods: This study was done over a period of 18 months in a tertiary care hospital. Two sets of cervical smears were prepared of which one was stained with conventional Pap stain, and other was stained with REAP stain. The smears were examined for cytomorphological parameters and were evaluated using a modification of parameters given by Ng et al. Results: A total of 737 smears were examined in duplicate. Most of the conventional Pap smears showed excellent preservation (91.6%) with very few showing optimal (7.6%) and sub-optimal staining (0.8%). In contrast to this excellent preservation was seen in just 33.6% of the REAP stained smears with majority showing optimal and sub-optimal preservation (46.5% and 20% respectively). The P value was statistically significant (<0.0001) depicting inferior staining quality of REAP stain. Conclusion: Rapid, economical, acetic acid Papanicolaou stain undoubtly is a simple, fast and cost effective stain which can be adopted mainly in resource limited settings, but cannot be utilized for research purpose in a tertiary care setup due to poor preservation of the staining quality. PMID:25538385

  9. Automated single-slide staining device. [in clinical bacteriology

    NASA Technical Reports Server (NTRS)

    Wilkins, J. R.; Mills, S. M.

    1975-01-01

    An automatic single-slide Gram staining device is described. A timer-actuated solenoid controls the dispensing of gentian violet, Gram iodine solution, decolorizer, and 1% aqueous safranin in proper sequence and for the time required for optimum staining. The amount of stain or reagent delivered is controlled by means of stopcocks below each solenoid. Used stains and reagents can be flushed automatically or manually. Smears Gram stained automatically are equal in quality to those prepared manually. The time to complete one Gram cycle is 4.80 min.

  10. Centrifuge-operated specimen staining method and apparatus

    NASA Technical Reports Server (NTRS)

    Clarke, Mark S. F. (Inventor); Feeback, Daniel L. (Inventor)

    1999-01-01

    A method of staining preselected, mounted specimens of either biological or nonbiological material enclosed within a staining chamber where the liquid staining reagents are applied and removed from the staining chamber using hypergravity as the propelling force. In the preferred embodiment, a spacecraft-operated centrifuge and method of diagnosing biological specimens while in orbit, characterized by hermetically sealing a shell assembly. The assembly contains slide stain apparatus with computer control therefor, the operative effect of which is to overcome microgravity, for example on board an International Space Station.

  11. Queer paradox/paradoxical queer: Anne Garréta's Pas un jour (2002).

    PubMed

    Cairns, Lucille

    2007-01-01

    This paper shows how Anne Garréta's Pas un jour (2002) is a decidedly queer text, in both the new and the old sense of that contested epithet. I examine three interrelated concerns central to Pas un jour. First, I analyze Garréta's mediation of desire in general: her own experiences of it; modalities thereof which subvert more 'normative' models of lesbianism; and her convergences with other gay, but male writers and theorists of desire such as Guy Hocquenghem, Gilles Deleuze and Michel Foucault. Second, I interrogate Garréta's dichotomy between desire and friendship, and adumbrate contrasts with Foucauldian theory. Finally, I scrutinize the meaning and value attributed to the particular body of desire with which Garréta is most commonly associated-homosexuality- and their links with those of a contemporary gay male writer, Dominique Fernandez. PMID:17804371

  12. Extracytoplasmic pas-like domains are common in signal transduction proteins. ;

    SciTech Connect

    Chang, C.; Tesar, C.; Gu, M.; Babnigg, G.; Joachimiak, A.; Pokkuluri, P. R.; Szurmant, H.; Schiffer, M.

    2010-02-01

    We present the crystal structure of the extracytoplasmic domain of the Bacillus subtilis PhoR sensor histidine kinase, part of a two-component system involved in adaptation to low environmental phosphate concentrations. In addition to the PhoR structure, we predict that the majority of the extracytoplasmic domains of B. subtilis sensor kinases will adopt a fold similar to the ubiquitous PAS domain.

  13. Extracytoplasmic PAS-like domains are common in signal transduction proteins.

    SciTech Connect

    Schiffer, M.; Chang, C.; Tesar, C.; Gu, M.; Babnigg, G.; Joachimiak, A.; Pokkuluri, P. R.; Szurmant, H.; Biosciences Division; Scripps Research Inst.

    2010-02-01

    We present the crystal structure of the extracytoplasmic domain of the Bacillus subtilis PhoR sensor histidine kinase, part of a two-component system involved in adaptation to low environmental phosphate concentrations. In addition to the PhoR structure, we predict that the majority of the extracytoplasmic domains of B. subtilis sensor kinases will adopt a fold similar to the ubiquitous PAS domain.

  14. The roles of primary care PAs and NPs caring for older adults with diabetes.

    PubMed

    Everett, Christine M; Thorpe, Carolyn T; Palta, Mari; Carayon, Pascale; Gilchrist, Valerie J; Smith, Maureen A

    2014-04-01

    Electronic health record data linked with Medicare data from an academic physician group were used to propose a multidimensional characterization of PA and NP roles on panels of primary care patients with diabetes. Seven PA and NP roles were defined based on level of involvement, visits with complex patients, and delivery of chronic care. Findings suggest that PAs and NPs in primary care perform a variety of roles and frequently perform multiple roles within a clinic. PMID:24662258

  15. Practical Analysis of materials with depth varying compositions using FT-IR photoacoustic spectroscopy (PAS)

    SciTech Connect

    J.F. McClelland; R.W. Jones; Siquan Luo

    2004-09-30

    FT-IR photoacoustic spectroscopy (PAS) is discussed as a nondestructive method to probe the molecular composition of materials versus depth on the basis of the analysis of layers of experimentally controllable thickness, which are measured from the sample surface to depths of some tens of micrometers, depending on optical and thermal properties. Computational methods are described to process photoacoustic amplitude and phase spectra for both semi-quantitative and quantitative depth analyses. These methods are demonstrated on layered and gradient samples.

  16. A cell cycle kinase with tandem sensory PAS domains integrates cell fate cues

    PubMed Central

    Mann, Thomas H.; Seth Childers, W.; Blair, Jimmy A.; Eckart, Michael R.; Shapiro, Lucy

    2016-01-01

    All cells must integrate sensory information to coordinate developmental events in space and time. The bacterium Caulobacter crescentus uses two-component phospho-signalling to regulate spatially distinct cell cycle events through the master regulator CtrA. Here, we report that CckA, the histidine kinase upstream of CtrA, employs a tandem-PAS domain sensor to integrate two distinct spatiotemporal signals. Using CckA reconstituted on liposomes, we show that one PAS domain modulates kinase activity in a CckA density-dependent manner, mimicking the stimulation of CckA kinase activity that occurs on its transition from diffuse to densely packed at the cell poles. The second PAS domain interacts with the asymmetrically partitioned second messenger cyclic-di-GMP, inhibiting kinase activity while stimulating phosphatase activity, consistent with the selective inactivation of CtrA in the incipient stalked cell compartment. The integration of these spatially and temporally regulated signalling events within a single signalling receptor enables robust orchestration of cell-type-specific gene regulation. PMID:27117914

  17. PAS/POLY-HAMP SIGNALING IN AER-2, A SOLUBLE HEME-BASED SENSOR

    PubMed Central

    Watts, Kylie J; Taylor, Barry L; Johnson, Mark S

    2011-01-01

    SUMMARY Poly-HAMP domains are widespread in bacterial chemoreceptors, but previous studies have focused on receptors with single HAMP domains. The Pseudomonas aeruginosa chemoreceptor, Aer-2, has an unusual domain architecture consisting of a PAS sensing domain sandwiched between three N-terminal and two C-terminal HAMP domains, followed by a conserved kinase control module. The structure of the N-terminal HAMP domains was recently solved, making Aer-2 the first protein with resolved poly-HAMP structure. The role of Aer-2 in P. aeruginosa is unclear, but here we show that Aer-2 can interact with the chemotaxis system of Escherichia coli to mediate repellent responses to oxygen, carbon monoxide and nitric oxide. Using this model system to investigate signaling and poly-HAMP function, we determined that the Aer-2 PAS domain binds penta-coordinated b-type heme and that reversible signaling requires four of the five HAMP domains. Deleting HAMP 2 and/or 3 resulted in a kinase-off phenotype, whereas deleting HAMP 4 and/or 5 resulted in a kinase-on phenotype. Overall, these data support a model in which ligand-bound Aer-2 PAS and HAMP 2 and 3 act together to relieve inhibition of the kinase control module by HAMP 4 and 5, resulting in the kinase-on state of the Aer-2 receptor. PMID:21255112

  18. Platelet storage in PAS-2 or autologous plasma: impact on functional parameters.

    PubMed

    Bunescu, A; Hild, M; Lundahl, J; Egberg, N

    2001-04-01

    Currently, several platelet additive solutions for long-term platelet storage have been introduced. The aim of this study was to compare the deterioration of functional status of platelets stored for up to 5 days in autologous plasma (AP) only, with platelet stored in PAS-2, a salt solution containing acetate, citrate and sodium chloride. Change in platelet adhesion, aggregation and activation was measured by flow cytometric technique. In addition, beta-Thromboglobulin (beta-TG), lactate and glucose were determined. After 5 days of storage, the expression of P-Selectin was significantly higher, the production of lactate and the consumption of glucose were significantly lower, in platelets stored in PAS-2 than in autologous plasma. No significant differences were detected on day 5 between the two groups with regard to fibrinogen, von Willebrand factor binding capacity, or to beta-TG release. It can be concluded that neither storage medium was consistently better for the parameters tested. However, it must be emphasized that platelets stored in autologous plasma exhibited less lesion, in terms of P-Selectin expression compared with platelets stored in PAS-2. PMID:11299027

  19. Laser diode bars based on strain-compensated AlGaPAs/GaAs heterostructures

    SciTech Connect

    Marmalyuk, Aleksandr A; Ladugin, M A; Yarotskaya, I V; Panarin, V A; Mikaelyan, G T

    2012-01-31

    Traditional (in the AlGaAs/GaAs system) and phosphorus-compensated (in the AlGaAs/AlGaPAs/GaAs system) laser heterostructures emitting at a wavelength of 850 nm are grown by MOVPE and studied. Laser diode bars are fabricated and their output characteristics are studied. The method used to grow heterolayers allowed us to control (minimise) mechanical stresses in the AlGaPAs/GaAs laser heterostructure, which made it possible to keep its curvature at the level of the initial curvature of the substrate. It is shown that the use of a compensated AlGaPAs/GaAs heterostructure improves the linear distribution of emitting elements in the near field of laser diode arrays and allows the power - current characteristic to retain its slope at high pump currents owing to a uniform contact of all emitting elements with the heat sink. The radius of curvature of the grown compensated heterostructures turns out to be smaller than that of traditional heterostructures.

  20. Allelic effects of mouse Pas1 candidate genes in human lung cancer cell lines.

    PubMed

    Galbiati, Federica; Pettinicchio, Angela; Dragani, Tommaso A; Manenti, Giacomo

    2006-12-01

    Four of the six genes constituting the mouse Pulmonary adenoma susceptibility 1 (Pas1) locus haplotype carry amino acid variants: Lrmp, Casc1, Ghiso, and Lmna-rs1. In vitro colony formation assay of human lung cancer cell lines A549 and NCI-H520 transfected with the allelic variants of the four genes revealed allele-specific modulations of colony numbers by Lmna-rs1 and Casc1, but not by Lrmp or Ghiso. In A549 and NCI-H520 cells, the A/J allele of Lmna-rs1 produced approximately 4- and approximately 2-fold, respectively, more transfectants than did the C57BL/6J allele, whereas the A/J allele of Casc1 produced approximately 6- and approximately 5-fold fewer transfectants, respectively, as compared to the C57BL/6J allele. Inhibition of clonogenicity by allelic forms of Pas1 candidate genes was not mediated by induction of apoptosis. These findings provide evidence that allelic variants of mouse Pas1 candidate genes differentially modulate growth of human cancer cells. PMID:16458428

  1. Principles of Ligand Binding within a Completely Buried Cavity in HIF2[alpha] PAS-B

    SciTech Connect

    Key, Jason; Scheuermann, Thomas H.; Anderson, Peter C.; Daggett, Valerie; Gardner, Kevin H.

    2010-04-19

    Hypoxia-inducible factors (HIFs) are heterodimeric transcription factors responsible for the metazoan hypoxia response and promote tumor growth, metastasis, and resistance to cancer treatment. The C-terminal Per-ARNT-Sim (PAS) domain of HIF2{alpha} (HIF2{alpha} PAS-B) contains a preformed solvent-inaccessible cavity that binds artificial ligands that allosterically perturb the formation of the HIF heterodimer. To better understand how small molecules bind within this domain, we examined the structures and equilibrium and transition-state thermodynamics of HIF2{alpha} PAS-B with several artificial ligands using isothermal titration calorimetry, NMR exchange spectroscopy, and X-ray crystallography. Rapid association rates reveal that ligand binding is not dependent upon a slow conformational change in the protein to permit ligand access, despite the closed conformation observed in the NMR and crystal structures. Compensating enthalpic and entropic contributions to the thermodynamic barrier for ligand binding suggest a binding-competent transition state characterized by increased structural disorder. Finally, molecular dynamics simulations reveal conversion between open and closed conformations of the protein and pathways of ligand entry into the binding pocket.

  2. A Study on N2O Measurement Characteristics Using Photoacoustic Spectroscopy (PAS)

    PubMed Central

    Kang, Soyoung; Kim, Seoungjin; Kang, Seongmin; Lee, Jeongwoo; Cho, Chang-Sang; Sa, Jea-Hwan; Jeon, Eui-Chan

    2014-01-01

    N2O, which is emitted mainly from nitrogen decomposition via bacteria, livestock manure, agricultural fertilizer use, fossil fuel combustion and waste incineration, is classified as a substance that causes significant destruction of the ozone layer. The N2O measurement methods for these emission sources may be divided into chromatography, optical, and electrical current measurements. Chromatography has been widely utilized for analyzing N2O. However, up until now, few studies have been conducted on N2O using photoacoustic spectroscopy. Therefore, this study aimed to evaluate performance of photoacoustic spectroscopy in this regard based on laboratory and field test results. The repeatability of photoacoustic spectroscopy was measured at 1.12%, which is lower than the repeatability of 3.0% suggested by the ISO 1564 standard, so, it has shown an excellent repeatability. The detection limit was determined to be 0.025 ppm, and the response time was confirmed to be 3 min and 26 s. The results of comparison between these measurements and GC show that the latter has superior accuracy, but mobility and convenience are superior for PAS. On the contrary, GC has a continuous measurement limitation, but PAS makes it possible to conduct continuous measurements. Therefore, PAS can be extremely useful to confirm the characteristics of N2O emissions and to quantify their amount. PMID:25106022

  3. PAS/poly-HAMP signalling in Aer-2, a soluble haem-based sensor.

    PubMed

    Watts, Kylie J; Taylor, Barry L; Johnson, Mark S

    2011-02-01

    Poly-HAMP domains are widespread in bacterial chemoreceptors, but previous studies have focused on receptors with single HAMP domains. The Pseudomonas aeruginosa chemoreceptor, Aer-2, has an unusual domain architecture consisting of a PAS-sensing domain sandwiched between three N-terminal and two C-terminal HAMP domains, followed by a conserved kinase control module. The structure of the N-terminal HAMP domains was recently solved, making Aer-2 the first protein with resolved poly-HAMP structure. The role of Aer-2 in P. aeruginosa is unclear, but here we show that Aer-2 can interact with the chemotaxis system of Escherichia coli to mediate repellent responses to oxygen, carbon monoxide and nitric oxide. Using this model system to investigate signalling and poly-HAMP function, we determined that the Aer-2 PAS domain binds penta-co-ordinated b-type haem and that reversible signalling requires four of the five HAMP domains. Deleting HAMP 2 and/or 3 resulted in a kinase-off phenotype, whereas deleting HAMP 4 and/or 5 resulted in a kinase-on phenotype. Overall, these data support a model in which ligand-bound Aer-2 PAS and HAMP 2 and 3 act together to relieve inhibition of the kinase control module by HAMP 4 and 5, resulting in the kinase-on state of the Aer-2 receptor. PMID:21255112

  4. [Parental alienation syndrome (PAS): unknown in medical settings, endemic in courts].

    PubMed

    Pignotti, Maria Serenella

    2013-02-01

    A purposed syndrome of so-called parental alienation (PAS), unsupported by any evidence-based data, unknown in medical settings, unquoted in medical books, absent in DSM and ICD, never demonstrated by controlled studies published in high scientific level journals, is rampant in Courts where it can lead to loose parental custody. During a divorce trial, almost always the mothers and the children, become joint in a sort of folie au deux, in a denigration campaign of ex-husband/father. From a review on this issue it seems evident its theoretical roots lie on a theory that justify gender violence and children sexual abuse. The bias that both of them are layers and that he children have not autonomy block their possibility of any defence in front of a Court. In severe cases, PAS becomes a new and efficient tool of intra-familiar violence. The treatment of severe cases is to stop any contact between mother and children. The resort to PAS in Courts must be strongly rejected. PMID:23535957

  5. A cell cycle kinase with tandem sensory PAS domains integrates cell fate cues.

    PubMed

    Mann, Thomas H; Seth Childers, W; Blair, Jimmy A; Eckart, Michael R; Shapiro, Lucy

    2016-01-01

    All cells must integrate sensory information to coordinate developmental events in space and time. The bacterium Caulobacter crescentus uses two-component phospho-signalling to regulate spatially distinct cell cycle events through the master regulator CtrA. Here, we report that CckA, the histidine kinase upstream of CtrA, employs a tandem-PAS domain sensor to integrate two distinct spatiotemporal signals. Using CckA reconstituted on liposomes, we show that one PAS domain modulates kinase activity in a CckA density-dependent manner, mimicking the stimulation of CckA kinase activity that occurs on its transition from diffuse to densely packed at the cell poles. The second PAS domain interacts with the asymmetrically partitioned second messenger cyclic-di-GMP, inhibiting kinase activity while stimulating phosphatase activity, consistent with the selective inactivation of CtrA in the incipient stalked cell compartment. The integration of these spatially and temporally regulated signalling events within a single signalling receptor enables robust orchestration of cell-type-specific gene regulation. PMID:27117914

  6. Enhancement of hERG channel activity by scFv antibody fragments targeted to the PAS domain.

    PubMed

    Harley, Carol A; Starek, Greg; Jones, David K; Fernandes, Andreia S; Robertson, Gail A; Morais-Cabral, João H

    2016-08-30

    The human human ether-à-go-go-related gene (hERG) potassium channel plays a critical role in the repolarization of the cardiac action potential. Changes in hERG channel function underlie long QT syndrome (LQTS) and are associated with cardiac arrhythmias and sudden death. A striking feature of this channel and KCNH channels in general is the presence of an N-terminal Per-Arnt-Sim (PAS) domain. In other proteins, PAS domains bind ligands and modulate effector domains. However, the PAS domains of KCNH channels are orphan receptors. We have uncovered a family of positive modulators of hERG that specifically bind to the PAS domain. We generated two single-chain variable fragments (scFvs) that recognize different epitopes on the PAS domain. Both antibodies increase the rate of deactivation but have different effects on channel activation and inactivation. Importantly, we show that both antibodies, on binding to the PAS domain, increase the total amount of current that permeates the channel during a ventricular action potential and significantly reduce the action potential duration recorded in human cardiomyocytes. Overall, these molecules constitute a previously unidentified class of positive modulators and establish that allosteric modulation of hERG channel function through ligand binding to the PAS domain can be attained. PMID:27516548

  7. Immunogold silver staining for light microscopy.

    PubMed

    Lackie, P M

    1996-07-01

    The immunogold silver staining method (IGSS) is widely used as a sensitive and specific immunohistochemical visualisation technique. IGSS involves the specific deposition of metallic silver at the site of immunogold labelling and provides a means of visualisation at low magnification by light or electron microscopy. Silver developers for IGSS rapidly deposit metallic silver only at the site of heavy metals, including gold and silver, because of their catalytic activity. The developing solution contains the silver ions and reducing agent necessary for this reaction. Using different silver salts as ion donors and by selecting an appropriate temperature and pH, visible amounts of silver can be deposited in a few minutes at the site of colloidal gold labelling while little non-specific background deposition occurs. Inclusion of protective colloids in the solution can also be used to control the reaction. Although studies of the chemical basis of silver deposition around unlabelled colloidal gold date back to 1939, immunogold enhancement by silver was established in 1983. The IGSS method evolved from the combination of disparate photographic, histochemical and immunogold techniques which have been effectively combined and optimised over the last 10 years to provide a visualisation system which is well suited to many immunohistochemical studies. PMID:8858363

  8. Safranine fluorescent staining of wood cell walls.

    PubMed

    Bond, J; Donaldson, L; Hill, S; Hitchcock, K

    2008-06-01

    Safranine is an azo dye commonly used for plant microscopy, especially as a stain for lignified tissues such as xylem. Safranine fluorescently labels the wood cell wall, producing green/yellow fluorescence in the secondary cell wall and red/orange fluorescence in the middle lamella (ML) region. We examined the fluorescence behavior of safranine under blue light excitation using a variety of wood- and fiber-based samples of known composition to interpret the observed color differentiation of different cell wall types. We also examined the basis for the differences in fluorescence emission using spectral confocal microscopy to examine lignin-rich and cellulose-rich cell walls including reaction wood and decayed wood compared to normal wood. Our results indicate that lignin-rich cell walls, such as the ML of tracheids, the secondary wall of compression wood tracheids, and wood decayed by brown rot, tend to fluoresce red or orange, while cellulose-rich cell walls such as resin canals, wood decayed by white rot, cotton fibers and the G-layer of tension wood fibers, tend to fluoresce green/yellow. This variation in fluorescence emission seems to be due to factors including an emission shift toward red wavelengths combined with dye quenching at shorter wavelengths in regions with high lignin content. Safranine fluorescence provides a useful way to differentiate lignin-rich and cellulose-rich cell walls without counterstaining as required for bright field microscopy. PMID:18802812

  9. Differential staining of interspecific chromosomes in somatic cell hybrids by alkaline Giemsa stain.

    PubMed

    Friend, K K; Chen, S; Ruddle, F H

    1976-03-01

    Staining of chromosome preparations of Chinese hamster-human hybrid cells and mouse-chimpanzee hybrids with alkaline Giemsa has yielded color differentiation of the interspecific chromosomes. Bicolor chromosomes, indicating apparent translocations also are observed for each of these hybrids. The specific color differences observed provide a rapid means of recognizing and aiding in the identification of the interspecific chromosomes and apparent translocations in these somatic cell hybrids. PMID:1028166

  10. Cytokeratin 7 staining in mammary and extramammary Paget's disease.

    PubMed

    Smith, K J; Tuur, S; Corvette, D; Lupton, G P; Skelton, H G

    1997-11-01

    There are a variety of routine and immunohistochemical stains used to diagnose mammary and extramammary Paget's disease (MPD and EMPD). Most of the stains commonly used, however, show a positive reaction in the Paget's cells in all cases. We wanted to assess which immunohistochemical stain is the best for the diagnosis of MPD and EMPD, as well as the best stain for identifying small foci of tumors in evaluating tumor margins. We evaluated nine cases of MPD and nine cases of EMPD, which were randomly chosen, with a battery of immunohistochemical stains. These stains included cytokeratin 7, cytokeratin 20, carcinoembryonic antigen, Ber-EP4, and CAM 5.2. Cytokeratin 7 was the only immunohistochemical stain that stained all of the cases diffusely, and, in all of the cases, the staining of the Paget's cell was intense and specific within the epidermis. We concluded that cytokeratin 7 is the immunohistochemical stain of choice in the diagnosis of Paget's disease. Because cytokeratin 7 seems to identify single cells, it might also be valuable in evaluating surgical margins for small foci in a tumor such as EMPD, which might have a multifocal origin. PMID:9388055

  11. Multi-class stain separation using independent component analysis

    NASA Astrophysics Data System (ADS)

    Trahearn, Nicholas; Snead, David; Cree, Ian; Rajpoot, Nasir

    2015-03-01

    Stain separation is the process whereby a full colour histology section image is transformed into a series of single channel images, each corresponding to a given stain's expression. Many algorithms in the field of digital pathology are concerned with the expression of a single stain, thus stain separation is a key preprocessing step in these situations. We present a new versatile method of stain separation. The method uses Independent Component Analysis (ICA) to determine a set of statistically independent vectors, corresponding to the individual stain expressions. In comparison to other popular approaches, such as PCA and NNMF, we found that ICA gives a superior projection of the data with respect to each stain. In addition, we introduce a correction step to improve the initial results provided by the ICA coefficients. Many existing approaches only consider separation of two stains, with primary emphasis on Haematoxylin and Eosin. We show that our method is capable of making a good separation when there are more than two stains present. We also demonstrate our method's ability to achieve good separation on a variety of different stain types.

  12. IgG Subclass Staining in Routine Renal Biopsy Material.

    PubMed

    Hemminger, Jessica; Nadasdy, Gyongyi; Satoskar, Anjali; Brodsky, Sergey V; Nadasdy, Tibor

    2016-05-01

    Immunofluorescence staining plays a vital role in nephropathology, but the panel of antibodies used has not changed for decades. Further classification of immunoglobulin (Ig)G-containing immune-type deposits with IgG subclass staining (IgG1, IgG2, IgG3, and IgG4) has been shown to be of diagnostic utility in glomerular diseases, but their value in the evaluation of renal biopsies has not been addressed systematically in large renal biopsy material. Between January 2007 and June 2014, using direct immunofluorescence, we stained every renal biopsy for the IgG subclasses if there was moderate to prominent glomerular IgG staining and/or IgG-predominant or IgG-codominant glomerular staining. The total number of biopsies stained was 1084, which included 367 cases of membranous glomerulonephritis, 307 cases of lupus nephritis, 74 cases of fibrillary glomerulonephritis, 53 cases of proliferative glomerulonephritis with monoclonal IgG deposits, and 25 cases of antiglomerular basement membrane disease, among others. We found that monoclonality of IgG deposits cannot always be reliably determined on the basis of kappa and lambda light chain staining alone, particularly if concomitant (frequently nonspecific) IgM staining is present. In IgG heavy and heavy and light chain deposition disease (3 cases), subclass staining is very helpful, and in proliferative glomerulonephritis with monoclonal IgG deposits subclass staining is necessary. IgG subclass staining is useful in differentiating primary from secondary membranous glomerulonephritis. In proliferative glomerulonephritis with polyclonal IgG deposition, IgG1 dominance/codominance with concomitant IgG3 and IgG2 but weak or absent IgG4 staining favors an underlying autoimmune disease. IgG subclass staining is a very useful diagnostic method in a selected cohort of renal biopsies, particularly in biopsies with glomerulonephritis with monoclonal IgG deposits. PMID:26848798

  13. Harmonization of the intracellular cytokine staining assay.

    PubMed

    Welters, Marij J P; Gouttefangeas, Cécile; Ramwadhdoebe, Tamara H; Letsch, Anne; Ottensmeier, Christian H; Britten, Cedrik M; van der Burg, Sjoerd H

    2012-07-01

    Active immunotherapy for cancer is an accepted treatment modality aiming to reinforce the T-cell response to cancer. T-cell reactivity is measured by various assays and used to guide the clinical development of immunotherapeutics. However, data obtained across different institutions may vary substantially making comparative conclusions difficult. The Cancer Immunotherapy Immunoguiding Program organizes proficiency panels to identify key parameters influencing the outcome of commonly used T-cell assays followed by harmonization. Our successes with IFNγ-ELISPOT and peptide HLA multimer analysis have led to the current study on intracellular cytokine staining (ICS). We report the results of three successive panels evaluating this assay. At the beginning, 3 out of 9 participants (33 %) were able to detect >6 out of 8 known virus-specific T-cell responses in peripheral blood of healthy individuals. This increased to 50 % of the laboratories in the second phase. The reported percentages of cytokine-producing T cells by the different laboratories were highly variable with coefficients of variation well over 60 %. Variability could partially be explained by protocol-related differences in background cytokine production leading to sub-optimal signal-to-noise ratios. The large number of protocol variables prohibited identification of prime guidelines to harmonize the assays. In addition, the gating strategy used to identify reactive T cells had a major impact on assay outcome. Subsequent harmonization of the gating strategy considerably reduced the variability within the group of participants. In conclusion, we propose that first basic guidelines should be applied for gating in ICS experiments before harmonizing assay protocol variables. PMID:22714399

  14. Visible luminescence from silicon wafers subjected to stain etches

    NASA Technical Reports Server (NTRS)

    Fathauer, R. W.; George, T.; Ksendzov, A.; Vasquez, R. P.

    1992-01-01

    Etching of Si in a variety of solutions is known to cause staining. These stain layers consist of porous material similar to that produced by anodic etching of Si in HF solutions. In this work, photoluminescence peaked in the red from stain-etched Si wafers of different dopant types, concentrations, and orientations produced in solutions of HF:HNO3:H2O was observed. Luminescence is also observed in stain films produced in solutions of NaNO2 in HF, but not in stain films produced in solutions of CrO3 in HF. The luminescence spectra are similar to those reported recently for porous Si films produced by anodic etching in HF solutions. However, stain films are much easier to produce, requiring no special equipment.

  15. Manual hematoxylin and eosin staining of mouse tissue sections.

    PubMed

    Cardiff, Robert D; Miller, Claramae H; Munn, Robert J

    2014-06-01

    The hematoxylin and eosin (H&E) stain is the standard used for microscopic examination of tissues that have been fixed, processed, embedded, and sectioned. It can be performed manually or by automation. For economic reasons, the manual technique is generally the method of choice for facilities with a low sample volume. This protocol describes manual H&E staining of fixed, processed, paraffin-embedded, and sectioned mouse tissues. In H&E-stained tissues, the nucleic acids stain dark blue and the proteins stain red to pink or orange. For accurate phenotyping and delineation of tissue detail, the protocol must be adhered to rigorously. This includes frequent reagent changes as well as the use of "in-date" reagents. Appropriate color in a good H&E stain allows for identification of many tissue subtleties that are necessary for accurate diagnosis. PMID:24890205

  16. Histochemical study of cutaneous mucins in hypothyroid dogs.

    PubMed

    Doliger, S; Delverdier, M; Moré, J; Longeart, L; Régnier, A; Magnol, J P

    1995-11-01

    A dermal mucinosis, visualized as dermal alcianophilic material, is occasionally present in canine hypothyroidism (myxedema). Various histochemical reactions (alcian blue/periodic acid-Schiff [PAS], alcian blue at pH 2.6, alcian blue at pH 1.0, critical electrolytical concentrations with and without dimethylsulfoxide, differential hydrolysis by hyaluronidases) were performed on skin biopsies from six dogs (four females and two males ranging from 8 to 13 years) affected by hypothyroidism, all of them presenting dermal mucinosis in hematoxylin and eosin-stained sections. In these dogs, the only polysaccharidic compound involved in the dermal mucinosis was hyaluronic acid. In this study, hyaluronic acid dermal deposits of hypothyroid dogs were significantly different from those of controls in subepidermal connective tissue and loose reticular connective tissue but not in periadnexal zones. We recommend the combined alcian blue/PAS reaction as a routine technique to assess dermal mucinosis in hypothyroid dogs. PMID:8592797

  17. Classification of abomasal displacement in cows according to histopathology of the liver and clinical chemistry.

    PubMed

    Komatsu, Y; Itoh, N; Taniyama, H; Kitazawa, T; Yokota, H; Koiwa, M; Ohtsuka, H; Terasaki, N; Maeno, K; Mizoguchi, M; Takeuchi, Y; Tanigawa, M; Nakamura, T; Watanabe, H; Matsuguchi, Y; Kukino, T; Honma, A

    2002-11-01

    Histopathological features of livers and blood chemical values in cows with abomasal displacement were investigated. Liver biopsy samples were collected during redressment operations in 92 cows with abomasal displacement, and the samples were stained with haematoxylin and eosin or periodic acid Schiff (PAS). Blood was collected for chemical tests. Livers were histopathologically divided into the following four types: normal histology cases (21%), fatty degeneration cases (36%), cloudy swelling cases (19%) and fatty degeneration cases with cloudy swelling (24%). The number of PAS-positive samples was significantly higher in the normal histology group and significantly lower in the severe fatty degeneration group and severe cloudy swelling group. Cows with fatty degeneration had significantly higher levels of serum 3-hydroxybutyric acid, non-esterified fatty acid and aspartate aminotransferase than did those with cloudy swelling or normal histology. The results indicate that the morbid conditions of cows with abomasal displacement can be classified into four types. PMID:12489873

  18. Bioactivity of crude ethanol extract and fractions of Eugenia uniflora (Myrtaceae) in the hepatopancreas of Oreochromis niloticus L.

    PubMed

    Fiuza, Tatiana S; Silva, Paulo C; De Paula, José R; Tresvenzol, Leonice M F; Sabóia-Morais, Simone M T

    2009-01-01

    This study evaluates the bioactivity of the crude ethanol extract and ethyl acetate, hexane and chloroform fractions obtained from Eugenia uniflora leaves using the hepatopancreas of Oreochromis niloticus L. as an experimental model. The ethanol extract and fractions were administered to the fish orally with their feed. Twenty-four hours later, the fish were sacrificed and their livers dissected, fixed in neutral formalin, embedded in paraffin and sectioned. Histological analyses were performed using Masson's trichrome and Haematoxylin-Eosin. Histochemical studies were performed using Feulgen, PAS (Periodic Acid Schiff) and PAS + salivary amylase and Sudan IV stain. The qualitative analysis of the material showed that the crude extract and the ethyl, chloroform and hexane fractions induced vasodilation, vascular congestion and toxicity due to the presence of eosinophilic granular cells, rodlet cells, some leukocytic infiltrate and rare focal necroses. The Nile tilapia proved to be a satisfactory model for screening plant products. PMID:20140296

  19. [Two cases of multi-drug-resistant pulmonary tuberculosis with para-aminosalicylic acid (PAS)-induced hypothyroidism].

    PubMed

    Mizutani, H; Horiba, M; Shindoh, J; Kimura, T; Son, M; Wakahara, K

    2001-10-01

    Two cases of multi-drug-resistant pulmonary tuberculosis with para-aminosalicylic acid (PAS)-induced hypothyroidism were reported. Case 1; a 73-year-old male, complaining of edema, was admitted to our hospital. He had been treated for his multi-drug-resistant pulmonary tuberculosis during the past 1 year with an antituberculous regimen consisting of ethambutol (EB), ethionamide (ETH) and PAS. A thyroid profile performed when he was admitted to our hospital showed several marked abnormalities: serum thyroid stimulating hormone (TSH) was elevated (69.4 microIU/ml: normal, 0.4-4.2 mIU/ml), free thyroxine level (T4) (0.01 ng/dl; normal, 0.70-1.60 ng/dl) and free triiodothyronine level (Ts) (0.60 pg/ml; normal, 2.3-4.1 pg/ml) were low. PAS was discontinued after he was admitted to our hospital, since PAS was believed to be the cause of the hypothyroidism. A thyroid profile that was repeated after the exclusion of PAS from treatment showed the following results: the TSH level was decreased (13.4 mIU/ml), the free T4 (0.93 ng/dl) were normal. During treatment with PAS, he had never received thyroid replacement therapy. Case 2; A 22-year-old female, complaining of hemosputum. She had been treated for her multi-drug-resistant pulmonary tuberculosis during the past 11 months with an antituberculous regimen consisting of EB, ETH and PAS. A thryoid profile performed when she was admitted to our hospital showed several marked abnormalities: elevated serum TSH (112.7 mIU/ml), and low T4 (2.0 micrograms/dl) and T3 (1.1 ng/ml). A thyroid profile that was repeated after the exclusion of PAS from treatment showed the following results: the TSH level was decreased (5.1 mIU/ml). Drug-induced hypothyroidism is an infrequent side effect of therapy with PAS, and only a few cases of PAS-induced hypothyroidism have been reported so far. In this report, we describe patients with hypothyroidism who were receiving therapy for multi-drug-resistant tuberculosis, tuberculosis namely, resistant

  20. Cutaneous myxidiosis in European eel, Anguilla anguilla (Linnaeus, 1758): histopathology, histochemistry and laminin immunohistochemistry.

    PubMed

    Manera, M; Borreca, C; Dezfuli, B S

    2016-07-01

    Histopathology, histochemistry and immunohistochemistry of the integument of European eel, Anguilla anguilla (Linnaeus, 1758), infected by Myxidium sp. are reported. Skin samples from affected and unaffected eels were dissected, formalin fixed, paraffin embedded, sectioned and stained with H&E, Periodic acid-Schiff's staining method, Alcian Blue 8 GX pH 2.5/Periodic acid-Schiff's and McCallum-Goodpasture's Gram stain. Moreover, immunohistochemistry was performed using a primary polyclonal laminin antibody. Histologically, cysts (diameter 2-3 mm) were observed mainly under the scale pockets, encircled by a thin collagen layer, lined by elongated, flattened fibroblasts and containing bipolar, PAS- and Gram-positive spores with opposite polar capsules. The epidermis stretched by the underlying cyst appeared dysplastic, thinned with a significant reduction in mucous cells number. Only inconsistent and aspecific inflammatory reaction was noted around the cysts at the dermis/epidermis interface. Intense laminin-like protein immunolabel was documented in the plasmodial ectoplasm and related to host anergia. This was the first report of laminin immunolabel in a member of the Myxozoa. Epidermal dysplasia represents likely an aspecific response against the underlying tensile force exerted by the developing parasite cyst, while fibroblast and collagen encapsulation denote a parasite-driven host response protecting, rather than harming, the encircled parasite. PMID:26525491

  1. The availability of the primer activation signal (PAS) affects the efficiency of HIV-1 reverse transcription initiation

    PubMed Central

    Ooms, Marcel; Cupac, Daniel; Abbink, Truus E. M.; Huthoff, Hendrik; Berkhout, Ben

    2007-01-01

    Initiation of reverse transcription of a retroviral RNA genome is strictly regulated. The tRNA primer binds to the primer binding site (PBS), and subsequent priming is triggered by the primer activation signal (PAS) that also pairs with the tRNA. We observed that in vitro reverse transcription initiation of the HIV-1 leader RNA varies in efficiency among 3′-end truncated transcripts, despite the presence of both PBS and PAS motifs. As the HIV-1 leader RNA can adopt two different foldings, we investigated if the conformational state of the transcripts did influence the efficiency of reverse transcription initiation. However, mutant transcripts that exclusively fold one or the other structure were similarly active, thereby excluding the possibility of regulation of reverse transcription initiation by the structure riboswitch. We next set out to determine the availability of the PAS element. This sequence motif enhances the efficiency of reverse transcription initiation, but its activity is regulated because the PAS motif is initially base paired within the wild-type template. We measured that the initiation efficiency on different templates correlates directly with accessibility of the PAS motif. Furthermore, changes in PAS are critical to facilitate a primer-switch to a new tRNA species, demonstrating the importance of this enhancer element. PMID:17308346

  2. Protective effects of ebselen (Ebs) and para-aminosalicylic acid (PAS) against manganese (Mn)-induced neurotoxicity

    SciTech Connect

    Marreilha dos Santos, A.P.; Lucas, Rui L.; Andrade, Vanda; Mateus, M. Luísa; Milatovic, Dejan; Aschner, Michael; Batoreu, M. Camila

    2012-02-01

    Chronic, excessive exposure to manganese (Mn) may induce neurotoxicity and cause an irreversible brain disease, referred to as manganism. Efficacious therapies for the treatment of Mn are lacking, mandating the development of new interventions. The purpose of the present study was to investigate the efficacy of ebselen (Ebs) and para-aminosalicylic acid (PAS) in attenuating the neurotoxic effects of Mn in an in vivo rat model. Exposure biomarkers, inflammatory and oxidative stress biomarkers, as well as behavioral parameters were evaluated. Co-treatment with Mn plus Ebs or Mn plus PAS caused a significant decrease in blood and brain Mn concentrations (compared to rats treated with Mn alone), concomitant with reduced brain E{sub 2} prostaglandin (PGE{sub 2}) and enhanced brain glutathione (GSH) levels, decreased serum prolactin (PRL) levels, and increased ambulation and rearing activities. Taken together, these results establish that both PAS and Ebs are efficacious in reducing Mn body burden, neuroinflammation, oxidative stress and locomotor activity impairments in a rat model of Mn-induced toxicity. -- Highlights: ► The manuscript is unique in its approach to the neurotoxicity of Mn. ► The manuscript incorporates molecular, cellular and functional (behavioral) analyses. ► Both PAS and Ebs are effective in restoring Mn behavioral function. ► Both PAS and Ebs are effective in reducing Mn-induced oxidative stress. ► Both PAS and Ebs led to a decrease in Mn-induced neuro-inflammation.

  3. Evaluation of existence region and formation time of particle accumulation structure (PAS) in half-zone liquid bridge

    NASA Astrophysics Data System (ADS)

    Gotoda, M.; Sano, T.; Kaneko, T.; Ueno, I.

    2015-03-01

    We focused on the particle accumulation structure (PAS) produced by the thermocapillary effect in a half-zone liquid bridge. Although models of the formation of the PAS have been previously proposed, they have not been experimentally verified. An assessment of the region in which the PAS exists is very subjective and often dependent on the observer, and this has necessitated the development of an objective and quantitative evaluation method. We therefore conducted a series of experiments to verify the physical model of the particle path lines in a rotating frame of reference using the fundamental frequency of the hydrothermal wave. We evaluated the intensity of the particle accumulation based on a modification of the "accumulation measure" proposed by Kuhlmann and Muldoon (Phys. Rev. E, 2012) to objectively and quantitatively determine the existence region of the SL-1 PAS. The results of the quantitative experiment revealed that the best aspect ratio (ratio of the height to radius) of the liquid bridge for the SL-1 PAS was about 0.64, and that the PAS formation time was nearly the same as the thermal diffusion time under the considered conditions (184 words, within 200 words).

  4. JPCam: Status of the 1.2Gpixel camera for the J-PAS survey.

    NASA Astrophysics Data System (ADS)

    Taylor, Keith; Marín-Franch, Antonio; Santoro, Fernando; Mendes de Oliveira, Claudia; Dupke, Renato

    2015-08-01

    JPCam is a 14-CCD mosaic camera, using the new e2v 9k-by-9k, 10 micron pixel detectors, to be deployed on the JST/T250, a dedicated 2.55m wide-field telescope at the OAJ (Observatorio Astrofísico de Javalambre) in Teruel, Spain. The camera is designed to perform the J-PAS, a BAO survey of the northern sky. The J-PAS survey strategy will use 59 filters, 56 relatively narrow-band (14.5nm) filters equi-spaced between 350 and 1000nm plus 3 broad-band filters to achieve unprecedented photometric red-shift accuracies for faint galaxies over 8000 square degrees of sky. The cryostat, detector mosaic and read electronics is being supplied by e2v. Each science detector is read from 16 ports simultaneously, allowing read times of 14s with a typical read noise of 5e- (rms). The JPCam focal plane will be complemented with 4 autoguiding and 8 wavefront sensing CCDs. Image quality over the whole CCD mosaic will be maintained by applying the required, close loop, focus, tilt and rotation adjustments to the focal plane through the JPCam hexapod actuator system, manufactured by NTE-Sener. The filter unit has been designed to admit 5 filter trays, each mounting 14 filters corresponding to the 14 CCDs of the mosaic. Each CCD will view only its corresponding filter avoiding optical cross-talk from their neighbors. With this configuration, JPCam will cover 4.7 square degrees with a plate scale of 0.2267 arcsec/pix and will allow all the J-PAS filters to be permanently installed on the camera. The massive 525mm aperture shutter is supplied by Bonn-Shutter UG.

  5. Burkholderia cepacia complex Phage-Antibiotic Synergy (PAS): antibiotics stimulate lytic phage activity.

    PubMed

    Kamal, Fatima; Dennis, Jonathan J

    2015-02-01

    The Burkholderia cepacia complex (Bcc) is a group of at least 18 species of Gram-negative opportunistic pathogens that can cause chronic lung infection in cystic fibrosis (CF) patients. Bcc organisms possess high levels of innate antimicrobial resistance, and alternative therapeutic strategies are urgently needed. One proposed alternative treatment is phage therapy, the therapeutic application of bacterial viruses (or bacteriophages). Recently, some phages have been observed to form larger plaques in the presence of sublethal concentrations of certain antibiotics; this effect has been termed phage-antibiotic synergy (PAS). Those reports suggest that some antibiotics stimulate increased production of phages under certain conditions. The aim of this study is to examine PAS in phages that infect Burkholderia cenocepacia strains C6433 and K56-2. Bcc phages KS12 and KS14 were tested for PAS, using 6 antibiotics representing 4 different drug classes. Of the antibiotics tested, the most pronounced effects were observed for meropenem, ciprofloxacin, and tetracycline. When grown with subinhibitory concentrations of these three antibiotics, cells developed a chain-like arrangement, an elongated morphology, and a clustered arrangement, respectively. When treated with progressively higher antibiotic concentrations, both the sizes of plaques and phage titers increased, up to a maximum. B. cenocepacia K56-2-infected Galleria mellonella larvae treated with phage KS12 and low-dose meropenem demonstrated increased survival over controls treated with KS12 or antibiotic alone. These results suggest that antibiotics can be combined with phages to stimulate increased phage production and/or activity and thus improve the efficacy of bacterial killing. PMID:25452284

  6. Jpcam: a 1.2 Gpixel Camera for the J-Pas Survey

    NASA Astrophysics Data System (ADS)

    Taylor, K.; Marín-Franch, A.; Laporte, R.; Santoro, F. G.; Marrara, L.; Cepa, J.; Cenarro, A. J.; Chueca, S.; Cristobal-Hornillos, D.; Ederoclite, A.; Gruel, N.; Moles, M.; Rueda, F.; Rueda, S.; Varela, J.; Yanes, A.; Benitez, N.; Dupke, R.; Fernández-Soto, A.; Jorden, P.; Lousberg, G.; Molino Benito, A.; Palmer, I.; Mendes de Oliveira, C.; Sodré, L.

    JPCam is a 14-CCD mosaic camera, using the new e2v 9k-by-9k 10 μm-pixel 16-channel detectors, to be deployed on a dedicated 2.55 m wide-field telescope at the OAJ (Observatorio Astrofísico de Javalambre) in Aragon, Spain. The camera is designed to perform a Baryon Acoustic Oscillations (BAO) survey of the northern sky. The J-PAS survey strategy will use 54 relatively narrow-band ( 13.8 nm) filters equi-spaced between 370 and 920 nm plus 3 broad-band filters to achieve unprecedented photometric red-shift accuracies for faint galaxies over 8000 square degrees of sky. The cryostat, detector mosaic and read electronics, is being supplied by e2v under contract to J-PAS while the mechanical structure, housing the shutter and filter assembly, is being designed and constructed by a Brazilian consortium led by INPE (Instituto Nacional de Pesquisas Espaciais). The cryostat is bridged to the telescope via a hexapod actuator system to maintain image quality across the field. Four sets of 14 filters are placed in the ambient environment, just above the dewar window but directly in line with the detectors, leading to a mosaic having 10 mm gaps between each CCD. The massive 500 mm aperture shutter is expected to be supplied by the Bonn-Shutter UG. We will present an overview of JPCam, from the filter configuration through to the CCD mosaic camera. A brief outline of the main J-PAS science projects will be included.

  7. Second Annual Maintenance, Inspection, and Test Report for PAS-1 Cask Certification for Shipping Payload B

    SciTech Connect

    KELLY, D.J.

    2000-10-09

    The Nuclear Packaging, Inc. (NuPac), PAS-1 cask is required to undergo annual maintenance and inspections to retain certification in accordance with U.S. Department of Energy (DOE) Certificate of Compliance USA/9184B(U) (Appendix A). The packaging configuration being tested and maintained is the NuPac PAS-1 cask for Payload B. The intent of the maintenance and inspections is to ensure the packaging remains in unimpaired physical condition. Two casks, serial numbers 2162-026 and 2162-027, were maintained, inspected, and tested at the 306E Development, Fabrication, and Test Laboratory, located at the Hanford Site's 300 Area. Waste Management Federal Services, Inc. (WMFS), a subsidiary of GTS Duratek, was in charge of the maintenance and testing. Cogema Engineering Corporation (Cogema) directed the operations in the test facility. The maintenance, testing, and inspections were conducted successfully with both PAS-1 casks. The work conducted on the overpacks included weighing, gasket replacement, and plastic pipe plug and foam inspections. The work conducted on the secondary containment vessel (SCV) consisted of visual inspection of the vessel and threaded parts (i.e., fasteners), visual inspection of sealing surfaces, replacement of O-ring seals, and a helium leak test. The work conducted on the primary containment vessel (PCV) consisted of visual inspection of the vessel and threaded parts (i.e., fasteners), visual inspection of sealing surfaces, replacement of O-ring seals, dimensional inspection of the vessel bottom, a helium leak test, and dye penetrant inspection of the welds. The vermiculite material used in the cask rack assembly was replaced.

  8. Comparative study of subculture, Gram staining and acridine orange staining for early detection of positive blood cultures.

    PubMed Central

    Mascart, G; Bertrand, F; Mascart, P

    1983-01-01

    In view of the importance of a rapid aetiological diagnosis in septicaemia, we compared the results of subculture, Gram staining and acridine orange staining in the detection of positive blood cultures. The study was based on 1013 blood cultures of which 138 were positive by culture. The three techniques were applied 12 h after the specimen was taken in 210 instances, at 24 h in 540 instances and after 48 h in 525. We were able to demonstrate the value of direct examination. Staining with acridine orange yields more positive results than Gram staining and is also simpler. PMID:6188764

  9. Comparison of tetrachromic VOF stain to other histochemical staining techniques for characterizing stromal soft and hard tissue components.

    PubMed

    Belaldavar, C; Hallikerimath, S; Angadi, P V; Kale, A D

    2014-11-01

    The components of hard tissues including dentin, enamel, cementum, bone and other calcified deposits, and mature and immature collagen pose problems for identification in routine hematoxylin and eosin (H & E) stained sections. Use of combinations of stains can demonstrate the components of hard tissues and soft tissues distinctly. We assessed the efficacy of the Verde Luz-orange G-acid fuchsin (VOF) stain for differentiating hard and soft connective tissues and compared results with other histochemical staining techniques. Eighty tissue sections comprising developing tooth (30), ossifying fibroma (30) and miscellaneous pathologies (20) expected to contain varying types of calcified tissues were stained with H & E, VOF, and Masson's trichrome (MT). In developing tooth, VOF demonstrated better differentiation of hard tissues, while it was comparable to MT for ossifying fibroma and miscellaneous pathologies. The intensity of staining was greater with VOF than with the other stains studied. VOF stains hard tissue components distinctly and gives good contrast with the surrounding connective tissue. VOF is comparable to MT, but has added advantages including single step staining, rapid and easy procedures, and it distinguishes the maturity of the tissues. PMID:24830362

  10. 7 CFR 3201.87 - Wood and concrete stains.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 15 2014-01-01 2014-01-01 false Wood and concrete stains. 3201.87 Section 3201.87... Designated Items § 3201.87 Wood and concrete stains. (a) Definition. Products that are designed to be applied as a finish for concrete and wood surfaces and that contain dyes or pigments to change the...

  11. 7 CFR 3201.87 - Wood and concrete stains.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 15 2013-01-01 2013-01-01 false Wood and concrete stains. 3201.87 Section 3201.87... Designated Items § 3201.87 Wood and concrete stains. (a) Definition. Products that are designed to be applied as a finish for concrete and wood surfaces and that contain dyes or pigments to change the...

  12. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and...

  13. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and...

  14. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and...

  15. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and...

  16. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and...

  17. The effect of selected staining techniques on bull sperm morphometry.

    PubMed

    Banaszewska, Dorota; Andraszek, Katarzyna; Czubaszek, Magdalena; Biesiada-Drzazga, Barbara

    2015-08-01

    Sperm morphometry has some value as an indicator of reproductive capacity in males. In laboratory practice a variety of slide-staining methods are used during morphological evaluation of semen to predict male fertility. The aim of this study was to determine the effect of staining of semen using four different techniques on the morphometry of the bull sperm cell. The material for the study consisted of semen collected from test bulls of the Black-and-White variety of Holstein-Friesians. The results obtained in the study indicate differences in the dimensions of bull sperm heads when different slide staining techniques were used. The most similar results for sperm head dimensions were obtained in the case of SpermBlue(®) and eosin+gentian violet complex, although statistically significant differences were found between all the staining techniques. Extreme values were noted for the other staining techniques - lowest for the Papanicolaou and highest for silver nitrate, which may indicate more interference in the cell by the reagents used in the staining process. However, silver nitrate staining was best at identifying the structures of the sperm cell. Hence it is difficult to determine which of the staining methods most faithfully reveals the dimensions and shape of the bull sperm. PMID:26149220

  18. In vivo photoacoustic imaging of model of port wine stains.

    PubMed

    Yuan, Kaihua; Yuan, Yi; Gu, Ying; Gao, Jianhua; Xing, Da

    2012-01-01

    Port wine stains are categorized as a benign capillary vascular malformation, which is hard to cure. In this paper, a photoacoustic microscopy system, which integrated a two-dimensional scanning galvanometer, an objective lens and a focused ultrasound transducer, was designed for noninvasive imaging of blood vessels of port wine stains model in vivo. Cock comb was chosen as the port wine stains model in the experiment. The blood vessels in x-y plane and x-z plane were imaged clearly. Experimental results demonstrate that photoacoustic microscopy can image the blood vessels of port wine stains model in vivo with high contrast and high resolution. It has the potential for clinical applications in detecting the blood vessels in port wine stains skin. PMID:22635179

  19. Effects of fixation, dehydration and staining on dimensions of myxosporidan.

    PubMed

    Parker, J D; Warner, M C

    1970-10-01

    The effects of fixation, dehydration and staining on the morphological dimensions of myxo- and microsporidan spores were tested. Seven fixatives, two dehydrants and five stains were tested. Ten % formalin produced the least shrinkage and provided the best cytological detail of fixed material in both types of spores. All fixatives caused shrinkage of myxosporidan spore length and polar capsule length. Spore capsule width and polar capsule width were unaffected by 10% formalin. Ethyl alcohol caused no significant change in spore width. Microsporidan spore length shrunk with all fixatives, but spore width was generally unaffected. Dehydration, with either isopropyl alcohol or acetone, produced additional, significant shrinkage. The influence of stains on spore size was negligible. Heidenhains iron hematoxylin followed by eosin, and Mallory's analine-blue collagen stain, effectively stained myxo- and microsporidan spores. PMID:16512155

  20. Staining and histomorphometry of microcracks in the human femoral head.

    PubMed

    Villanueva, A R; Longo, J A; Weiner, G

    1994-03-01

    We developed staining techniques that permit identification and histomorphometric analysis of microcracks in the human femoral head 1) from thick, ground bone sections (100 microns) by prestaining with the Villanueva mineralized bone stain (MIBS), and 2) from plastic embedded, undecalcified thin bone sections (5-15 microns) by staining in gallocyanin chrome alum-Villanueva blood stain methods. Both methods represent a significant improvement in the stainability of the microcracks, cellular and tissue elements, and the simultaneous assessment of osteoid seams and tetracycline markers by histomorphometry. Shrinkage and other artifacts were minimized, which helped to clarify some of the uncertainties arising from artifacts resulting from some bone staining methods. Histomorphometric analyses of microcracks were conducted on thick, ground sections of subchondral and trabecular bone. Microcracks were more prevalent in the subchondral bone and osteochondral junction than in the more distant trabeculae. We have consistently localized microcrack areas in bone tissues prepared in these ways. PMID:7515700

  1. Formation of the peroxisome lumen is abolished by loss of Pichia pastoris Pas7p, a zinc-binding integral membrane protein of the peroxisome.

    PubMed Central

    Kalish, J E; Theda, C; Morrell, J C; Berg, J M; Gould, S J

    1995-01-01

    We have cloned and sequenced PAS7, a gene required for peroxisome assembly in the yeast Pichia pastoris. The product of this gene, Pas7p, is a member of the C3HC4 superfamily of zinc-binding proteins. Point mutations that alter conserved residues of the C3HC4 motif abolish PAS7 activity and reduce zinc binding, suggesting that Pas7p binds zinc in vivo and that zinc binding is essential for PAS7 function. As with most pas mutants, pas7 cells exhibit a pronounced deficiency in import of peroxisomal matrix proteins that contain either the type 1 peroxisomal targeting signal (PTS1) or the type 2 PTS (PTS2). However, while other yeast and mammalian pas mutants accumulate ovoid, vesicular peroxisomal intermediates, loss of Pas7p leads to accumulation of membrane sheets and vesicles which lack a recognizable lumen. Thus, Pas7p appears to be essential for protein translocation into peroxisomes as well as formation of the lumen of the organelle. Consistent with these data, we find that Pas7p is an integral peroxisomal membrane protein which is entirely resistant to exogenous protease and thus appears to reside completely within the peroxisome. Our observations suggest that the function of Pas7p defines a previously unrecognized step in peroxisome assembly: formation of the peroxisome lumen. Furthermore, because the peroxisomal intermediates in the pas7 delta mutant proliferate in response to peroxisome-inducing environmental conditions, we conclude that Pas7p is not required for peroxisome proliferation. PMID:7565793

  2. Inhibitory PAS domain protein is a negative regulator of hypoxia-inducible gene expression

    NASA Astrophysics Data System (ADS)

    Makino, Yuichi; Cao, Renhai; Svensson, Kristian; Bertilsson, Göran; Asman, Mikael; Tanaka, Hirotoshi; Cao, Yihai; Berkenstam, Anders; Poellinger, Lorenz

    2001-11-01

    Alteration of gene expression is a crucial component of adaptive responses to hypoxia. These responses are mediated by hypoxia-inducible transcription factors (HIFs). Here we describe an inhibitory PAS (Per/Arnt/Sim) domain protein, IPAS, which is a basic helix-loop-helix (bHLH)/PAS protein structurally related to HIFs. IPAS contains no endogenous transactivation function but demonstrates dominant negative regulation of HIF-mediated control of gene expression. Ectopic expression of IPAS in hepatoma cells selectively impairs induction of genes involved in adaptation to a hypoxic environment, notably the vascular endothelial growth factor (VEGF) gene, and results in retarded tumour growth and tumour vascular density in vivo. In mice, IPAS was predominantly expressed in Purkinje cells of the cerebellum and in corneal epithelium of the eye. Expression of IPAS in the cornea correlates with low levels of expression of the VEGF gene under hypoxic conditions. Application of an IPAS antisense oligonucleotide to the mouse cornea induced angiogenesis under normal oxygen conditions, and demonstrated hypoxia-dependent induction of VEGF gene expression in hypoxic corneal cells. These results indicate a previously unknown mechanism for negative regulation of angiogenesis and maintenance of an avascular phenotype.

  3. Topological control of the Caulobacter cell cycle circuitry by a polarized single-domain PAS protein

    PubMed Central

    Sanselicio, Stefano; Bergé, Matthieu; Théraulaz, Laurence; Radhakrishnan, Sunish Kumar; Viollier, Patrick H.

    2015-01-01

    Despite the myriad of different sensory domains encoded in bacteria, only a few types are known to control the cell cycle. Here we use a forward genetic screen for Caulobacter crescentus motility mutants to identify a conserved single-domain PAS (Per-Arnt-Sim) protein (MopJ) with pleiotropic regulatory functions. MopJ promotes re-accumulation of the master cell cycle regulator CtrA after its proteolytic destruction is triggered by the DivJ kinase at the G1-S transition. MopJ and CtrA syntheses are coordinately induced in S-phase, followed by the sequestration of MopJ to cell poles in Caulobacter. Polarization requires Caulobacter DivJ and the PopZ polar organizer. MopJ interacts with DivJ and influences the localization and activity of downstream cell cycle effectors. Because MopJ abundance is upregulated in stationary phase and by the alarmone (p)ppGpp, conserved systemic signals acting on the cell cycle and growth phase control are genetically integrated through this conserved single PAS-domain protein. PMID:25952018

  4. A novel source of MeV positron bunches driven by energetic protons for PAS application

    NASA Astrophysics Data System (ADS)

    Tan, Zongquan; Xu, Wenzhen; Liu, Yanfen; Xiao, Ran; Kong, Wei; Ye, Bangjiao

    2014-11-01

    This paper proposes a novel methodology of MeV positrons generation for PAS application. Feasibility of this proposal analyzed by G4Beamline and Transport have shown reasonable success. Using 2 Hz, 1.6 GeV, 100 ns and 1.5 μC/bunch proton bunches for bombarding a graphite target, about 100 ns e+ bunches are generated. Quasi-monochromatic positrons in the range of 1-10 MeV included in these bunches have a flux of >107/s, peak brightness of 1014/s. A magnetic-confinement beamline is utilized to transport the positrons and a "Fast Beam Chopper" is unprecedentedly extended to chop those relativistic bunches. The positron beam can be finally characterized by the energy range of 1-10 MeV and bunch width from one hundred ps up to 1 ns. Such ultrashort bunches can be useful in tomography-type positron annihilation spectroscopy (PAS) as well as other applications.

  5. Molecular cloning, expression and immunological characterisation of Pas n 1, the major allergen of Bahia grass Paspalum notatum pollen.

    PubMed

    Davies, Janet M; Mittag, Diana; Dang, Thanh D; Symons, Karen; Voskamp, Astrid; Rolland, Jennifer M; O'Hehir, Robyn E

    2008-12-01

    Bahia grass, Paspalum notatum, is a clinically important subtropical grass with a prolonged pollination season from spring to autumn. We aimed to clone and characterise the major Bahia grass pollen allergen, Pas n 1. Grass pollen-allergic patients presenting to a tertiary hospital allergy clinic were tested for IgE reactivity with Bahia grass pollen extract by skin prick testing, ImmunoCAP, ELISA and immunoblotting. Using primers deduced from the N-terminal peptide sequence of a group 1 allergen of Bahia grass pollen extract separated by two-dimensional gel electrophoresis, the complete Pas n 1 cDNA was obtained by rapid amplification of cDNA ends and cloned. Biological relevance of recombinant Pas n 1 expressed in Escherichia coli was assessed by serum IgE reactivity and basophil activation. Twenty-nine of 34 (85%) consecutive patients presenting with grass pollen allergy were skin prick test positive to Bahia grass pollen. The Pas n 1 cDNA has sequence homology with the beta-expansin 1 glycoprotein family and is more closely related to the maize pollen group 1 allergen (85% identity) than to ryegrass Lol p 1 or Timothy grass Phl p 1 (64 and 66% identity, respectively). rPas n 1 reacted with serum IgE in 47 of 55 (85%) Bahia grass pollen-allergic patients, activated basophils and inhibited serum IgE reactivity with the 29 kDa band of Bahia grass pollen extract. In conclusion the cDNA for the major group 1 allergen of the subtropical Bahia grass pollen, Pas n 1, was identified and cloned. rPas n 1 is immunologically active and is a valuable reagent for diagnosis and specific immunotherapy of grass pollen allergy. PMID:18817975

  6. Application of FTIR-PAS and Raman spectroscopies for the determination of organic matter in farmland soils.

    PubMed

    Xing, Zhe; Du, Changwen; Tian, Kang; Ma, Fei; Shen, Yazhen; Zhou, Jianmin

    2016-09-01

    In soil analysis, Raman spectroscopy is not as widely used as infrared spectroscopy mainly owing to fluorescence interferences. This paper investigated the feasibility of Fourier-transform infrared photoacoustic (FTIR-PAS) and Raman spectroscopies for predicting soil organic matter (SOM) using partial least squares regression (PLSR) analysis. 194 farmland soil samples were collected and scanned with FTIR and Raman spectrometers in the spectral range of 4000-400cm(-1) and 180-3200cm(-1), respectively. For the PLSR models, the combined dataset was split into 146 samples as the calibration set (75%) and 48 samples as the validation set (25%). The optimal number of analytical factors was determined using a leave-one-out cross-validation. The results showed that SOM could be predicted using FTIR-PAS and Raman spectroscopies independently, with R(2)>0.70 and RPD>1.8 for the validation sets. In comparison to the single applications of FTIR-PAS and Raman spectroscopies, accurate prediction of SOM was made by combining FTIR-PAS and Raman spectroscopies, with R(2)=0.81 and RPD=2.18 for the validation sets. By statistically assessing large amounts of PLS models, model-population analysis confirmed that the accuracy of the PLS model can be increased by combining FTIR-PAS and Raman spectroscopies. In conclusion, the combination of FTIR-PAS and Raman spectroscopies is a promising alternative for soil characterization, especially for the prediction of SOM, owing to the availability of complementary information from both FTIR-PAS (polar vibrations) and Raman spectroscopy (non-polar vibrations). PMID:27343604

  7. Automated detection of cells from immunohistochemically-stained tissues: application to Ki-67 nuclei staining

    NASA Astrophysics Data System (ADS)

    Cinar Akakin, Hatice; Kong, Hui; Elkins, Camille; Hemminger, Jessica; Miller, Barrie; Ming, Jin; Plocharczyk, Elizabeth; Roth, Rachel; Weinberg, Mitchell; Ziegler, Rebecca; Lozanski, Gerard; Gurcan, Metin N.

    2012-03-01

    An automated cell nuclei detection algorithm is described to be used for the quantification of immunohistochemicallystained tissues. Detection and segmentation of positively stained cells and their separation from the background and negatively-stained cells is crucial for fast, accurate, consistent and objective analysis of pathology images. One of the major challenges is the identification, hence accurate counting of individual cells, when these cells form clusters. To identify individual cell nuclei within clusters, we propose a new cell nuclei detection method based on the well-known watershed segmentation, which can lead to under- or over-segmentation for this problem. Our algorithm handles oversegmentation by combining H-minima transformed watershed algorithm with a novel region merging technique. To handle under-segmentation problem, we develop a Laplacian-of-Gaussian (LoG) filtering based blob detection algorithm, which estimates the range of the scales from the image adaptively. An SVM classifier was trained in order to separate non-touching single cells and touching cell clusters with five features representing connected region properties such as eccentricity, area, perimeter, convex area and perimeter-to-area ratio. Classified touching cell clusters are segmented with the H-minima based watershed algorithm. The resulting over-segmented regions are improved with the merging algorithm. The remaining under-segmented cell clusters are convolved with LoG filters to detect the cells within them. Cell-by-cell nucleus detection performance is evaluated by comparing computer detections with cell locations manually marked by eight pathology residents. The sensitivity is 89% when the cells are marked as positive at least by one resident and it increases to 99% when the evaluated cells are marked by all eight residents. In comparison, the average reader sensitivity varies between 70% +/- 18% and 95% +/- 11%.

  8. Novel Process for Laser Stain Removal from Archaeological Oil Paintings

    NASA Astrophysics Data System (ADS)

    El-Nadi, Lotfia; El-Feky, Osama; Abdellatif, Galila; Darwish, Sawsan

    2013-03-01

    Some samples of oil paintings (5 × 5 cm) were prepared on wooden panel with four types of fungi commonly encountered on oil paintings were selected for this study. Each of the fungi is associated with different colored stains. Fungus Alternaria tenuis is associated by a dense black stain, Chetomium globosum by a brownish gray stain, Aspergillus flavus by a yellowish stain, and Fusaruim oxysporum by a pinkish stain. Fungi growing on oil paintings affect the surface characteristics by forming a variety of colored patches typically composed of many complex chemical substances that are produced during metabolic processes. These colored stains may be encrusted in spores, present in mycelium or secreted to a substance such as oil paintings surfaces. While the fungal stains can sometimes be extracted with appropriate solvents, there are some stains that resist solvent extraction entirely. Developing new solvent system that might attack the paint structure, and is time consuming and requires a great deal of trial and error. Mechanical stain removal is also problematic in that it often produces abrasion of the surface, markedly deteriorating the artwork, and is extra ordinarily fine and tedious. For these reasons, we decided to examine an alternative physical technique as a new approach to deal with stain removal. Since the stains are due to the existence of fungi, we thought it a good idea to remove them by singlet oxygen. We applied the photo dynamic process through which the fungi stains were covered with organic dye derivatives in solution under controlled illumination in the lab. The samples were then irradiated by low power Laser light from a He-Ne laser, the dye will be photodecomposed and produce singlet oxygen. We report in this work the results obtained as a function of: - The concentration and types of the organic dye in solution, - The presence of certain amounts of liquids added to the solution, - The scanning speed of the laser beam on the sample surface

  9. Confusion over live/dead stainings for the detection of vital microorganisms in oral biofilms - which stain is suitable?

    PubMed Central

    2014-01-01

    Background There is confusion over the definition of the term “viability state(s)” of microorganisms. “Viability staining” or “vital staining techniques” are used to distinguish live from dead bacteria. These stainings, first established on planctonic bacteria, may have serious shortcomings when applied to multispecies biofilms. Results of staining techniques should be compared with appropriate microbiological data. Discussion Many terms describe “vitality states” of microorganisms, however, several of them are misleading. Authors define “viable” as “capable to grow”. Accordingly, staining methods are substitutes, since no staining can prove viability. The reliability of a commercial “viability” staining assay (Molecular Probes) is discussed based on the corresponding product information sheet: (I) Staining principle; (II) Concentrations of bacteria; (III) Calculation of live/dead proportions in vitro. Results of the “viability” kit are dependent on the stains’ concentration and on their relation to the number of bacteria in the test. Generally this staining system is not suitable for multispecies biofilms, thus incorrect statements have been published by users of this technique. To compare the results of the staining with bacterial parameters appropriate techniques should be selected. The assessment of Colony Forming Units is insufficient, rather the calculation of Plating Efficiency is necessary. Vital fluorescence staining with Fluorescein Diacetate and Ethidium Bromide seems to be the best proven and suitable method in biofilm research. Regarding the mutagenicity of staining components users should be aware that not only Ethidium Bromide might be harmful, but also a variety of other substances of which the toxicity and mutagenicity is not reported. Summary – The nomenclature regarding “viability” and “vitality” should be used carefully. – The manual of the commercial “viability” kit itself points out that

  10. Western Blot of Stained Proteins from Dried Polyacrylamide Gels

    NASA Technical Reports Server (NTRS)

    Gruber, Claudia; Stan-Lotter, Helga

    1996-01-01

    Western blotting of proteins is customarily performed following their separation on polyacrylamide gels, either prior to staining (1) or, as recently reported, following staining (2). We describe here Western blotting with stained gels, which had been dried and some of which had been stored for years. This procedure permits immunological analysis of proteins, to which antisera may have become available only later, or where the application of newly developed sensitive detection methods is desired. Once rehydration of the gels is achieved, proteins can be-transferred to blotting membranes by any appropriate protocol. Proteins stained with Coomassie Blue have to be detected with a non-chromogenic method, such as the film-based enhanced chemiluminescence (ECL)2) procedure (3). Silver stained proteins, which transfer in the colorless form, may be visualized by any detection method, although, because of the usually very low amounts of proteins, detection by ECL is preferable. Blotting of stained proteins from rehydrated gels is as rapid and as quantitative as from freshly prepared gels, in contrast to blotting from wet stained gels, which requires extensive washing and results in low transfer efficiency (2). Together with a photographic record of the gel pattern, unambiguous identification of immunoreactive proteins from complex mixtures is possible. Some further applications of this work are discussed.

  11. Histochemical and Biometric Study of the Gastrointestinal System of Hyla Orientalis (Bedriaga, 1890) (Anura, Hylidae)

    PubMed Central

    Akat, E.; Arıkan, H.; Göçmen, B.

    2014-01-01

    This study was carried out to assess the localization of hyaluronic acid (HA) and the distribution of glycoproteins in the gastrointestinal system of adult Hyla orientalis. Histochemical analysis of the gastrointestinal system in H. orientalis showed that mucous content included glycogene and/or oxidable dioles [periodic acid/Schiff (PAS)+], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS+) and acid sulphate [Aldehyde fuchsin (AF)+] glycoproteins. However the mucus content was not the same in stomach, small and large intestine. The mucus content of stomach included only glycogene and/or oxidable dioles and sialic acid residues. Besides these histochemical methods, the localization of HA was detected using biotinylated hyaluronic acid binding protein labeled with streptavidin-fluorescein isothiocyanate (FITC). In the extracellular matrix of the submucosa, the reaction for HA was evident. Since HA was located in submucosa beneath the epithelial layer of gastrointestinal system, it has a significant role in hydric balance, and essential to provide the gastrointestinal system integrity and functionality. According to biometric results, there were statistical differences between small and large intestine in terms of the amount of material stained positive with PAS/AB, PAS, KOH/PAS and AF/AB. Additionally, number of goblet cells in the small and large intestine was significantly different. PMID:25578977

  12. Use of immunohistochemical staining panel for characterisation of ovarian neoplasms.

    PubMed Central

    Ashorn, P; Helle, M; Helin, H; Ashorn, R; Krohn, K

    1988-01-01

    Eighty five ovarian epithelial and non-epithelial tumours were studied by peroxidase histochemical staining for their reactivity with six monoclonal human milk fat globule (HMFG) antibodies, peanut agglutinin (PNA) lectin, and a monoclonal cytokeratin antibody. HMFG IIIC12 and cytokeratin antibodies distinguished epithelial from non-epithelial tumours. The staining patterns of mucinous and serous tumours were essentially different from each other; poorly differentiated anaplastic carcinomas showed similar antigenic content to that of the serous cystadenocarcinomas. Furthermore, staining with PNA lectin and HMFG antibodies was useful in distinguishing clear cell carcinomas from other malignant epithelial tumours of the ovary. Images Fig 2 Fig 1 PMID:2449464

  13. Histochemical observations of the haemocytes of Locusta migratoria.

    PubMed

    Costin, N M

    1975-01-01

    Five of the six categories of haemocytes of Locusta migratoria, that is, the plasmatocytes, spherule cells, granulocytes, coagulocytes and oenocytoids, contain conspicuous granules of mucosubstance in their cytoplasm. The mucosubstance has been characterized by using a series of histochemical tests, including Alcian Blue staining at different pH levels and salt concentrations, the periodic acid-Schiff (PAS) test, the high iron diamine test, enzymatic digestions and sequential staining methods. The results indicate that four different mucosubstances occur in a granular form, although not all four are found in every blood cell type. The mucosubstances are a neutral glycoprotein and neuraminidase-resistant, sulphated and non-sulphated sialomucins. The non-sulphated sialomucin occurs in both periodate-reactive and -unreactive forms. PMID:47851

  14. Granular cell leiomyosarcoma of the skin

    SciTech Connect

    Suster, S.; Rosen, L.B.; Sanchez, J.L.

    1988-06-01

    A case is presented of a multifocal malignant neoplasm involving the skin of the upper back in a 10-year-old boy following radiation therapy to the head and neck for a cerebellar medulloblastoma. Histologically, the neoplastic cells were remarkable for the presence of abundant periodic acid-Schiff (PAS)-positive diastase-resistant intracytoplasmic eosinophilic granules. Immunoperoxidase procedures revealed strong positive staining of the tumor cells with desmin, vimentin, and smooth muscle myosin antibodies, and negative staining for myoglobin, S-100 protein and keratin, thus supporting a smooth muscle line of differentiation for this neoplasm. Electronmicroscopy demonstrated numerous intracytoplasmic autophagic vacuoles that corresponded to the granules observed under the light microscope. Leiomyosarcoma should be entertained in the differential diagnosis of poorly differentiated cutaneous neoplasms histologically characterized by a proliferation of cells containing abundant granular eosinophilic cytoplasm.

  15. [Evaluation of the website of the Nord-Pas-de-Calais regional pharmacovigilance center].

    PubMed

    Rochoy, Michaël; Béné, Johana; Messaadi, Nassir; Auffret, Marine; Gautier, Sophie

    2016-06-01

    We posted the Nord-Pas-de-Calais regional pharmacovigilance center website and distributed a survey to its potential users between August 2014 and October 2014 (135 general practitioners, 45 pharmacists, 14 patients). Satisfaction was 7.3±1.6 out of 10 points for the visual aspect, 7.8±1.5 out of 10 points for navigation and 7.6±1.4 out of 10 points for content. The website was declared useful by 98% respondents, particularly for the reporting of adverse drugs reactions (89%). PMID:27235657

  16. Steinway piano and stained glass clerestory window in lounge area, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Steinway piano and stained glass clerestory window in lounge area, upper deck. Hot water radiators can be seen at base of wall. These run throughout the houseboat. - Houseboat LA DUCHESSE, The Antique Boat Museum, Clayton, Jefferson County, NY

  17. 18. INTERIOR DETAIL VIEW OF STAINED GLASS WINDOW LOCATED AT ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    18. INTERIOR DETAIL VIEW OF STAINED GLASS WINDOW LOCATED AT SOUTH SIDE OF ALTAR, NOTE INSCRIPTION DEDICATED IN THE MEMORY OF FATHER DAMIEN - St. Francis Catholic Church, Moloka'i Island, Kalaupapa, Kalawao County, HI

  18. VIEW OF THREE SOUTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    VIEW OF THREE SOUTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ARE LOCATED ADJACENT TO THE ALTER. - U.S. Naval Base, Pearl Harbor, Chapel, Corner of Oakley & Nimitz Street, Pearl City, Honolulu County, HI

  19. Interior, detail closeup shot of window with stained glass inserts ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Interior, detail closeup shot of window with stained glass inserts in top southeast room taken from ther west - J. Weingartner & Son Cigar Factory, 414 East Walnut Street, North Wales, Montgomery County, PA

  20. 6. Vick Farm, interior perspective of stained glass window, added ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    6. Vick Farm, interior perspective of stained glass window, added as part of deck addition on west side. - Vick Farm, North side Idlewild Road, 0.2 mile northwest of Idlewild & Maplewood Drive, Burlington, Boone County, KY

  1. VIEW OF THREE NORTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    VIEW OF THREE NORTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ARE LOCATED ADJACENT TO THE ALTAR. - U.S. Naval Base, Pearl Harbor, Chapel, Corner of Oakley & Nimitz Street, Pearl City, Honolulu County, HI

  2. 18. INTERIOR OF KITCHEN NO. 1 SHOWING STAINED CABINETRY ON ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    18. INTERIOR OF KITCHEN NO. 1 SHOWING STAINED CABINETRY ON OPPOSITE WALL FROM PAINTED CABINETS. VIEW TO NORTHEAST. - Bishop Creek Hydroelectric System, Plant 6, Cashbaugh-Kilpatrick House, Bishop Creek, Bishop, Inyo County, CA

  3. VIEW OF THREE NORTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    VIEW OF THREE NORTH FACING STAINED GLASS WINDOWS. THESE WINDOWS ARE LOCATED JUST BELOW THE CHOIR LOFT. - U.S. Naval Base, Pearl Harbor, Chapel, Corner of Oakley & Nimitz Street, Pearl City, Honolulu County, HI

  4. INTERIOR VIEW OF ENTRY. SHOWING THE STAINED CONCRETE FLOOR AND ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    INTERIOR VIEW OF ENTRY. SHOWING THE STAINED CONCRETE FLOOR AND WINDOW WITH DIAMOND PATTERN MUNTINS. VIEW FACING NORTHWEST. - Hickam Field, Officers' Housing Type F, 602 Beard Avenue, Honolulu, Honolulu County, HI

  5. 4. September 1969 DETAIL OF STAINED GLASS WINDOWS IN EAST ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    4. September 1969 DETAIL OF STAINED GLASS WINDOWS IN EAST WALL, INTERIOR VIEW FROM BALCONY - Mount Zion United Methodist Church, 1334 Twenty-ninth Street Northwest, Washington, District of Columbia, DC

  6. INTERIOR VIEW OF THE LANAI. SHOWING THE ORIGINAL STAINED CONCRETE ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    INTERIOR VIEW OF THE LANAI. SHOWING THE ORIGINAL STAINED CONCRETE FLOOR WITH INCISED LINES, AND HINGED DOOR TO GARAGE WITH VERTICAL BOARD PANELING (BACKGROUND). VIEW FACING NORTHWEST. - Hickam Field, Officers' Housing Type J, 701 Beard Street, Honolulu, Honolulu County, HI

  7. The Type Ia Supernova Pipeline for the Javalambre Physics of the Accelerating Universe Astrophysical Survey (J-PAS)

    NASA Astrophysics Data System (ADS)

    Siffert, B. B.; Reis, R. R. R.; Calvão, M. O.

    2014-10-01

    The Javalambre Physics of the Accelerating Universe Astrophysical Survey (J-PAS) is an astronomical facility being built in Sierra de Javalambre, Spain. The main goal is to study the expansion of the Universe through different cosmological observables such as baryonic acoustic oscillations, type Ia supernovae and galaxy clusters. The main instrument will be a 2.5 m telescope equipped with a system of 56 narrow band filters in the optical. Here we present a sketch of the pipeline we are developing to detect type Ia supernovae with J-PAS. First we describe each individual step of the pipeline, such as image subtraction and source selection. Then we show some results we obtained when applying our pipeline to images from the Sloan Digital Sky Survey and the ALHAMBRA survey, which had a set of narrow band filters similar to the ones that will be used by J-PAS.

  8. A PAS domain-containing regulator controls flagella-flagella interactions in Campylobacter jejuni

    PubMed Central

    Reuter, Mark; Periago, Paula M.; Mulholland, Francis; Brown, Helen L.; van Vliet, Arnoud H. M.

    2015-01-01

    The bipolar flagella of the foodborne bacterial pathogen Campylobacter jejuni confer motility, which is essential for virulence. The flagella of C. jejuni are post-translationally modified, but how this process is controlled is not well understood. In this work, we have identified a novel PAS-domain containing regulatory system, which modulates flagella-flagella interactions in C. jejuni. Inactivation of the cj1387c gene, encoding a YheO-like PAS6 domain linked to a helix-turn-helix domain, resulted in the generation of a tightly associated “cell-train” morphotype, where up to four cells were connected by their flagella. The morphotype was fully motile, resistant to vortexing, accompanied by increased autoagglutination, and was not observed in aflagellated cells. The Δcj1387c mutant displayed increased expression of the adjacent Cj1388 protein, which comprises of a single endoribonuclease L-PSP domain. Comparative genomics showed that cj1387c (yheO) orthologs in bacterial genomes are commonly linked to an adjacent cj1388 ortholog, with some bacteria, including C. jejuni, containing another cj1388-like gene (cj0327). Inactivation of the cj1388 and cj0327 genes resulted in decreased autoagglutination in Tween-20-supplemented media. The Δcj1388 and Δcj0327 mutants were also attenuated in a Galleria larvae-based infection model. Finally, substituting the sole cysteine in Cj1388 for serine prevented Cj1388 dimerization in non-reducing conditions, and resulted in decreased autoagglutination in the presence of Tween-20. We hypothesize that Cj1388 and Cj0327 modulate post-translational modification of the flagella through yet unidentified mechanisms, and propose naming Cj1387 the Campylobacter Flagella Interaction Regulator CfiR, and the Cj1388 and Cj0327 protein as CfiP and CfiQ, respectively. PMID:26284050

  9. Immunohistochemical CD3 staining detects additional patients with celiac disease

    PubMed Central

    Mubarak, Amani; Wolters, Victorien M; Houwen, Roderick HJ; ten Kate, Fiebo JW

    2015-01-01

    AIM: To investigate whether performing immunohistochemical CD3 staining, in order to improve the detection of intra-epithelial lymphocytosis, has an additional value in the histological diagnosis of celiac disease. METHODS: Biopsies obtained from 159 children were stained by hematoxylin and eosin (HE) and evaluated using the Marsh classification. CD3 staining was subsequently evaluated separately and independently. RESULTS: Differences in evaluation between the routine HE sections and CD3 staining were present in 20 (12.6%) cases. In 10 (6.3%) patients the diagnosis of celiac disease (Marsh II and III) changed on examination of CD3 staining: in 9 cases, celiac disease had initially been missed on the HE sections, while 1 patient had been over-diagnosed on the routine sections. In all patients, the final diagnosis based on CD3 staining, was concordant with serological results, which was not found previously. In the other 10 (12.3%) patients, the detection of sole intra-epithelial lymphocytosis (Marsh I) improved. Nine patients were found to have Marsh I on CD3 sections, which had been missed on routine sections. Interestingly, the only patient with negative serology had Giardiasis. Finally, in 1 patient with negative serology, in whom Marsh I was suspected on HE sections, this diagnosis was withdrawn after evaluation of the CD3 sections. CONCLUSION: Staining for CD3 has an additional value in the histological detection of celiac disease lesions, and CD3 staining should be performed when there is a discrepancy between serology and the diagnosis made on HE sections. PMID:26140002

  10. Interior detail view, surviving stained glass panel in an east ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    Interior detail view, surviving stained glass panel in an east aisle window. Most of the stained glass has been removed from the building and relocated to other area churches. (Similar to HABS No. PA-6694-25). - Acts of the Apostles Church in Jesus Christ, 1400-28 North Twenty-eighth Street, northwest corner of North Twenty-eighth & Master Streets, Philadelphia, Philadelphia County, PA

  11. News from the Biological Stain Commission, No. 17.

    PubMed

    Lyon, H O

    2016-01-01

    In the 17(th) issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission's International Affairs Committee presents information from the 20(th) meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on October 15 - 17, 2014 in Toronto, Canada, and from the 29(th) meeting of CEN/TC 140 In vitro diagnostic medical devices held on February 3, 2015 in Berlin, Germany. PMID:26942571

  12. Protein Alpha Shape (PAS) Dock: a new gaussian-based score function suitable for docking in homology modelled protein structures.

    PubMed

    Tøndel, Kristin; Anderssen, Endre; Drabløs, Finn

    2006-03-01

    Protein Alpha Shape (PAS) Dock is a new empirical score function suitable for virtual library screening using homology modelled protein structures. Here, the score function is used in combination with the geometry search method Tabu search. A description of the protein binding site is generated using gaussian property fields like in Protein Alpha Shape Similarity Analysis (PASSA). Gaussian property fields are also used to describe the ligand properties. The overlap between the receptor and ligand hydrophilicity and lipophilicity fields is maximised, while minimising steric clashes. Gaussian functions introduce a smoothing of the property fields. This makes the score function robust against small structural variations, and therefore suitable for use with homology models. This also makes it less critical to include protein flexibility in the docking calculations. We use a fast and simplified version of the score function in the geometry search, while a more detailed version is used for the final prediction of the binding free energies. This use of a two-level scoring makes PAS-Dock computationally efficient, and well suited for virtual screening. The PAS-Dock score function is trained on 218 X-ray structures of protein- ligand complexes with experimental binding affinities. The performance of PAS-Dock is compared to two other docking methods, AutoDock and MOE-Dock, with respect to both accuracy and computational efficiency. According to this study, PAS-Dock is more computationally efficient than both AutoDock and MOE-Dock, and gives a better prediction of the free energies of binding. PAS-Dock is also more robust against structural variations than AutoDock. PMID:16652207

  13. Chromatin and Cell Wall Staining of Schizosaccharomyces pombe.

    PubMed

    Hagan, Iain M

    2016-01-01

    Fission yeasts grow by tip extension, maintaining a constant width until they reach a critical size threshold and divide. Division by medial fission-which gives these yeast their name-generates a new end that arises from the site of cytokinesis. The old end, which was produced during the previous cell cycle, initiates progression of the new cell cycle, and in G2, the new end is activated in a process termed new-end takeoff (NETO). In this protocol, the fluorescent stains calcofluor and 4',6-diamidino-2-phenylindole (DAPI) are used to give a rapid and informative assessment of morphogenesis and cell-cycle progression in the fission yeast Schizosaccharomyces pombe Calcofluor reveals the timing of NETO because it stains the birth scars that are generated at new ends by cytokinesis less efficiently than the rest of the cell wall. Intense calcofluor staining of the septum and measurement of cell length are also widely used to identify dividing cells and to gauge the timing of mitotic commitment. Staining nuclei with DAPI identifies mono- and binucleated cells and complements the calcofluor staining procedure to evaluate the stages of the cell cycle and identify mitotic errors. Equally simple DAPI staining procedures reveal chromatin structure in higher resolution, facilitating more accurate staging of mitotic progression and characterization of mitotic errors. PMID:27250942

  14. Black stain and dental caries: a review of the literature.

    PubMed

    Żyła, Tomasz; Kawala, Beata; Antoszewska-Smith, Joanna; Kawala, Maciej

    2015-01-01

    Black stain is characterized as a dark line or an incomplete coalescence of dark dots localized on the cervical third of the tooth. Over the last century, the etiology of black stain has been the subject of much debate. Most of the studies concerning this issue were conducted in pediatric population. According to the reviewed articles published between 2001 and 2014, the prevalence of black stain varies from 2.4% to 18% with equal sex distribution. The majority of the authors confirm the correlation between the presence of black stain and lower caries experience. The microflora of this deposit is dominated by Actinomyces spp. and has lower cariogenic potential than nondiscolored dental plaque. Iron/copper and sulfur complexes are thought to be responsible for the dark color. In patients with black stain saliva has higher calcium concentrations and higher buffering capacity. Factors such as dietary habits, socioeconomic status, and iron supplementation may be contributing to the formation of black stain. PMID:25802850

  15. Black Stain and Dental Caries: A Review of the Literature

    PubMed Central

    Żyła, Tomasz; Kawala, Beata; Antoszewska-Smith, Joanna; Kawala, Maciej

    2015-01-01

    Black stain is characterized as a dark line or an incomplete coalescence of dark dots localized on the cervical third of the tooth. Over the last century, the etiology of black stain has been the subject of much debate. Most of the studies concerning this issue were conducted in pediatric population. According to the reviewed articles published between 2001 and 2014, the prevalence of black stain varies from 2.4% to 18% with equal sex distribution. The majority of the authors confirm the correlation between the presence of black stain and lower caries experience. The microflora of this deposit is dominated by Actinomyces spp. and has lower cariogenic potential than nondiscolored dental plaque. Iron/copper and sulfur complexes are thought to be responsible for the dark color. In patients with black stain saliva has higher calcium concentrations and higher buffering capacity. Factors such as dietary habits, socioeconomic status, and iron supplementation may be contributing to the formation of black stain. PMID:25802850

  16. Functional anatomy of the lacrimal gland in African black ostrich Struthio camelus domesticus in the embryonic and postnatal period.

    PubMed

    Klećkowska-Nawrot, Joanna; Goździewska-Harłajczuk, Karolina; Nowaczyk, Renata; Krasucki, Krzysztof

    2015-01-01

    The aim of the present study was morphological and histochemical analysis of the lacrimalgland (LG) in African black ostrich Struthio camelus domesticus in the embryonic and postnatalperiod. Studies were conducted on 50 ostriches aged between the 28th day of incubation until7 months old. Tissue sections were stained with haematoxylin and eosin, Azan trichrome,periodic acid-Schiff, Alcian blue pH 2.5, aldehyde fuchsin and Hale's dialysed iron. The LGin ostrich was classified as a tubulo-acinar type. The primordia of the lobes were determinedin the LG structure on the 28th day of incubation, whilst the weakly visible lobes with aciniand tubules were observed on the 40th day of incubation. Morphometric studies of the LGshowed steady growth, characterised by an increase in both length and width. Histometricmeasurements of lobe size showed little difference between the first, second and third agegroups, whilst in the fourth age group a marked increase in size of lobes was observed.The study showed that, apart from morphological changes, during the growth of the LGthe character of acid mucopolysaccharides changed. Sulphated acid mucopolysaccharideswere indicated, particularly with aldehyde fuchsin (AF) staining in the fourth age group.The Hale's dialysed iron (HDI) staining showed a low concentration of carboxylated acidmucopolysaccharides in the first and second age groups and a higher concentration in thethird and fourth age groups. Periodic acid-Schiff staining (PAS)-positive cells were observedin each age group, but only a small number of cells with a weakly PAS-positive reaction weredemonstrated in the first age group. PMID:26017903

  17. PAS-Positive Extracellular Deposits within Germinal Centers of Hyperplastic Follicles during SIV infection in a Rhesus Macaque

    PubMed Central

    Hong, Jung Joo; Villinger, Francois; Courtney, Cynthia L

    2014-01-01

    Background Lymphoid tissue remodeling is characteristic of chronic SIV infection. Methods A rhesus macaque infected with SIVmac239 was necropsied and its lymphoid tissues subjected to histopathology characterization. Results Germinal centers in spleen and lymph nodes contained PAS-positive, non-amyloid extracellular deposits, decreased T follicular helper cells, and normal density of Ki67+ B cells. Conclusions A possible mechanism for PAS-positive deposits includes exaggerated involution of SIV-induced follicular hyperplasia secondary to virus-associated immune reaction. PMID:24628065

  18. Light and electron microscopic study of the eyelids, conjunctiva-associated lymphoid tissue and lacrimal gland in Bilgorajska Goose (Anser anser).

    PubMed

    Klećkowska-Nawrot, Joanna; Nowaczyk, Renata; Goździewska-Harłajczuk, Karolina; Barszcz, Karolina; Kowalczyk, Artur; Łukaszewicz, Ewa

    2016-01-01

    Normal structure of the accessory organs of the eye is essential for normal eye physiology. Among the most important accessory organs of the eye are the eyelids, the conjunctiva-associated lymphoid tissue (CALT) and the lacrimal gland (LG). The aim of this study was to demonstrate the histological structure of the eyelids and LG by histochemical and ultrastructural analysis. The study was performed on 13 adult female Bilgorajska geese. Eyelid samples were stained with the Alcian blue (AB pH 2.5) and periodic acid-Schiff (PAS) methods. Staining methods used for LG were AB pH 2.5, aldehyde fuchsin (AF), PAS and Hale's dialysed iron (HDI). Within the connective tissue of the eyelids, well-developed, diffuse, CALT follicles were observed, mostly under the conjunctival epithelium. Numerous lymphocytes were present within loose connective tissue. Staining of the eyelids with the PAS method demonstrated the presence of goblet cells of a mucous nature, and AB pH 2.5 staining indicated the presence of sulfated acid mucopolysaccharides. PAS staining of LG revealed the presence of secretory cells containing weakly PAS-positive granules. All epithelial cells of the corpus glandulae and the duct systems reacted positively to AB pH 2.5. HDI staining detected the presence of carboxylated acid mucopolysaccharides. Transmission electron microscopy investigations revealed two types of secretory epithelial cells in LG. Both types of LG cells contained drop-like secretory vesicles of different sizes with low or high electron density in cytoplasm, as well as small and large lipid vacuoles, and numerous small primary lysosomes. PMID:25673221

  19. Immunosuppressive PAS-1 is an excretory/secretory protein released by larval and adult worms of the ascarid nematode Ascaris suum.

    PubMed

    Antunes, M F P; Titz, T O; Batista, I F C; Marques-Porto, R; Oliveira, C F; Alves de Araujo, C A; Macedo-Soares, M F

    2015-05-01

    Helminths use several strategies to evade and/or modify the host immune response, including suppression or inactivation of the host antigen-specific response. Several helminth immunomodulatory molecules have been identified. Our studies have focused on immunosuppression induced by the roundworm Ascaris suum and an A. suum-derived protein named protein 1 from A. suum (PAS-1). Here we assessed whether PAS-1 is an excretory/secretory (E/S) protein and whether it can suppress lipopolysaccharide-induced inflammation. Larvae from infective eggs were cultured in unsupplemented Dulbecco's modified Eagle medium (DMEM) for 2 weeks. PAS-1 was then measured in the culture supernatants and in adult A. suum body fluid at different time points by enzyme-linked immunosorbent assay (ELISA) with the monoclonal antibody MAIP-1. Secreted PAS-1 was detected in both larval culture supernatant and adult body fluid. It suppressed lipopolysaccharide (LPS)-induced leucocyte migration and pro-inflammatory cytokine production, and stimulated interleukin (IL)-10 secretion, indicating that larval and adult secreted PAS-1 suppresses inflammation in this model. Moreover, the anti-inflammatory activity of PAS-1 was abolished by treatment with MAIP-1, a PAS-1-specific monoclonal antibody, confirming the crucial role of PAS-1 in suppressing LPS-induced inflammation. These findings demonstrate that PAS-1 is an E/S protein with anti-inflammatory properties likely to be attributable to IL-10 production. PMID:24703095

  20. Mouse D1Pas1, a DEAD-box RNA helicase, is required for the completion of first meiotic prophase in male germ cells.

    PubMed

    Inoue, Hiroki; Ogonuki, Narumi; Hirose, Michiko; Hatanaka, Yuki; Matoba, Shogo; Chuma, Shinichiro; Kobayashi, Kimio; Wakana, Shigeharu; Noguchi, Junko; Inoue, Kimiko; Tanemura, Kentaro; Ogura, Atsuo

    2016-09-16

    D1Pas1 is a mouse autosomal DEAD-box RNA helicase expressed predominantly in the testis. To assess its possible function, we generated D1Pas1-deficient mice using embryonic stem cells with a targeted D1Pas1 allele. Deletion of D1Pas1 did not cause noticeable embryonic defects or death, indicating that D1Pas1 is not essential for embryogenesis. Whereas homozygous knockout female mice showed normal reproductive performance, homozygous knockout male mice were completely sterile. The seminiferous epithelium of D1Pas1-deficient males contained no spermatids or spermatozoa because of spermatogenic arrest at the late pachytene stage. Upregulation of retrotransposons such as LINE-1 was not found in D1Pas1-deficient males, unlike males lacking Mvh, another testicular DEAD-box RNA helicase. Meiotic chromosome behavior in developing spermatocytes of D1Pas1-deficient males was indistinguishable from that in wild-type males, at least until synaptonemal complex formation. Thus, mouse D1Pas1 is the first-identified DEAD-box RNA helicase that plays critical roles in the final step of the first meiotic prophase in male germ cells. PMID:27473657

  1. Detection of Acid Fast Bacilli in Saliva using Papanicolaou Stain Induced Fluorescence Method Versus Fluorochrome Staining: An Evaluative Study

    PubMed Central

    (Munot), Priya P Lunawat; Mhapuskar, Amit A; Ganvir, S M; Hazarey, Vinay K; Mhapuskar, Madhavi A; Kulkarni, Dinraj

    2015-01-01

    Background: Fifty years after effective chemotherapy, tuberculosis (TB) still remains leading infectious cause of adult mortality. The aim of present study was to evaluate diagnostic utility of papanicolaou (Pap) stain induced fluorescence microscopic examination of salivary smears in the diagnosis of pulmonary TB. Materials and Methods: Cross-sectional study of 100 individuals clinically suspected of suffering from active pulmonary TB. Control group – 50 individuals are suffering from any pulmonary disease other than TB such as pneumonia or bronchiogenic carcinoma. Fluorescence microscopic examination of two salivary smears stained by Pap stain and auramine-rhodamine (A-R) stain respectively for each patient. Ziehl–Neelsen stained sputum smear examined under the light microscope for each patient. Culture was done in all the patients for microbiological confirmation. McNemar's Chi-square analysis, Kappa test, and Z-test. Results: The sensitivities of the three staining methods using culture as a reference method were 93.02%, 88.37% and 87.20% for Pap, A-R and Ziehl–Neelson respectively. Conclusion: Pap-induced fluorescence of salivary smears is a safe, reliable and rapid method, which can prove as a valuable diagnostic tool for diagnosis of TB. PMID:26229384

  2. Reliability of a rapid hematology stain for sputum cytology*

    PubMed Central

    Gonçalves, Jéssica; Pizzichini, Emilio; Pizzichini, Marcia Margaret Menezes; Steidle, Leila John Marques; Rocha, Cristiane Cinara; Ferreira, Samira Cardoso; Zimmermann, Célia Tânia

    2014-01-01

    Objective: To determine the reliability of a rapid hematology stain for the cytological analysis of induced sputum samples. Methods: This was a cross-sectional study comparing the standard technique (May-Grünwald-Giemsa stain) with a rapid hematology stain (Diff-Quik). Of the 50 subjects included in the study, 21 had asthma, 19 had COPD, and 10 were healthy (controls). From the induced sputum samples collected, we prepared four slides: two were stained with May-Grünwald-Giemsa, and two were stained with Diff-Quik. The slides were read independently by two trained researchers blinded to the identification of the slides. The reliability for cell counting using the two techniques was evaluated by determining the intraclass correlation coefficients (ICCs) for intraobserver and interobserver agreement. Agreement in the identification of neutrophilic and eosinophilic sputum between the observers and between the stains was evaluated with kappa statistics. Results: In our comparison of the two staining techniques, the ICCs indicated almost perfect interobserver agreement for neutrophil, eosinophil, and macrophage counts (ICC: 0.98-1.00), as well as substantial agreement for lymphocyte counts (ICC: 0.76-0.83). Intraobserver agreement was almost perfect for neutrophil, eosinophil, and macrophage counts (ICC: 0.96-0.99), whereas it was moderate to substantial for lymphocyte counts (ICC = 0.65 and 0.75 for the two observers, respectively). Interobserver agreement for the identification of eosinophilic and neutrophilic sputum using the two techniques ranged from substantial to almost perfect (kappa range: 0.91-1.00). Conclusions: The use of Diff-Quik can be considered a reliable alternative for the processing of sputum samples. PMID:25029648

  3. The stain prevention efficacy of two tooth whitening dentifrices.

    PubMed

    Ayad, Farid; De Sciscio, Peter; Stewart, Bernal; De Vizio, William; Petrone, Margaret E; Volpe, Anthony R

    2002-08-01

    An 8-week randomized, double-blind, parallel group clinical study was conducted to assess the extrinsic stain prevention efficacy of three commercially available dentifrices: 1) a dentifrice containing 0.243% sodium fluoride with copolymer, tetrasodium pyrophosphate, and sodium tripolyphosphate in a silica base (Product 1); 2) a dentifrice containing 0.243% sodium fluoride, baking soda and peroxide, tetrasodium pyrophosphate, and sodium tripolyphosphate in a silica base (Product 2); and 3) a dentifrice containing 0.243% sodium fluoride in a silica base (Product 3). After the collection of baseline stain scores by a trained examiner and a subsequent oral prophylaxis, 126 volunteers were randomized to one of the three treatment groups (balanced for composite extrinsic stain scores). Throughout the 8-week treatment period, subjects brushed their teeth twice daily with their assigned dentifrice. At baseline, 4-, and 8-week evaluations, extrinsic dental stain was measured on the facial surfaces of the six maxillary anterior teeth and on the facial and lingual surfaces of the six mandibular anterior teeth using the Lobene Index. A total of 120 subjects completed the study. No adverse events were reported, and subjects who discontinued the study did so for reasons unrelated to the dentifrices. At the 4-week evaluation, composite stain scores were statistically significantly lower (P < .05) for both Product 1 (44.9%) and for Product 2 (34.6%) relative to Product 3. At the 8-week evaluation, composite stain scores were statistically significantly lower (P < .05) for both Product 1 (28.4%) and for Product 2 (29.6%) relative to Product 3. The results of this clinical study demonstrate that both dentifrices, one containing 0.234% sodium fluoride with copolymer, tetrasodium pyrophosphate, and sodium tripolyphosphate in a silica base; and one with 0.243% sodium fluoride with copolymer, tetrasodium pyrophosphate, and sodium tripolyphosphate in a silica base are more effective in

  4. VITRAIL: Acquisition, Modeling, and Rendering of Stained Glass.

    PubMed

    Thanikachalam, Niranjan; Baboulaz, Loic; Prandoni, Paolo; Trumpler, Stefan; Wolf, Sophie; Vetterli, Martin

    2016-10-01

    Stained glass windows are designed to reveal their powerful artistry under diverse and time-varying lighting conditions; virtual relighting of stained glass, therefore, represents an exceptional tool for the appreciation of this age old art form. However, as opposed to most other artifacts, stained glass windows are extremely difficult if not impossible to analyze using controlled illumination because of their size and position. In this paper, we present novel methods built upon image based priors to perform virtual relighting of stained glass artwork by acquiring the actual light transport properties of a given artifact. In a preprocessing step, we build a material-dependent dictionary for light transport by studying the scattering properties of glass samples in a laboratory setup. We can now use the dictionary to recover a light transport matrix in two ways: under controlled illuminations the dictionary constitutes a sparsifying basis for a compressive sensing acquisition, while in the case of uncontrolled illuminations the dictionary is used to perform sparse regularization. The proposed basis preserves volume impurities and we show that the retrieved light transport matrix is heterogeneous, as in the case of real world objects. We present the rendering results of several stained glass artifacts, including the Rose Window of the Cathedral of Lausanne, digitized using the presented methods. PMID:27416590

  5. Methods of biological dosimetry employing chromosome-specific staining

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel

    2000-01-01

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.

  6. Methods And Compositions For Chromosome-Specific Staining

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel

    2003-08-19

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.

  7. Language Fluency and the Evaluation of Cultural Faux Pas: Russians Interviewing for Jobs in the United States

    ERIC Educational Resources Information Center

    Molinsky, Andrew

    2005-01-01

    How are nonnatives evaluated when committing cultural faux pas, and how does their fluency in the language of the foreign culture affect the evaluation of their culturally inappropriate behavior? I address these questions in the context of Russian professionals learning to interview for jobs in the United States, an arena of strong cultural…

  8. Biological control of Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) using Aeromonas phage PAS-1.

    PubMed

    Kim, J H; Choresca, C H; Shin, S P; Han, J E; Jun, J W; Park, S C

    2015-02-01

    The potential control efficacy of Aeromonas phage PAS-1 was evaluated against Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) model in this study. The phage was co-cultured with the virulent A. salmonicida subsp. salmonicida strain AS05 that possesses the type III secretion system (TTSS) ascV gene, and efficient bacteriolytic activity was observed against the bacteria. The administration of PAS-1 in rainbow trout demonstrated that the phage was cleared from the fish within 200 h post-administration, and a temporal neutralizing activity against the phage was detected in the sera of phage-administrated fish. The administration of PAS-1 (multiplicity of infection: 10 000) in A. salmonicida subsp. salmonicida infected rainbow trout model showed notable protective effects, with increased survival rates and mean times to death. These results demonstrated that Aeromonas phage PAS-1 could be considered as an alternative biological control agent against A. salmonicida subsp. salmonicida infections in rainbow trout culture. PMID:23594036

  9. Safety evaluation for packaging for the transport of K Basin sludge samples in the PAS-1 cask

    SciTech Connect

    SMITH, R.J.

    1998-11-17

    This safety evaluation for packaging authorizes the shipment of up to two 4-L sludge samples to and from the 325 Lab or 222-S Lab for characterization. The safety of this shipment is based on the current U.S. Department of Energy Certification of Compliance (CoC) for the PAS-1 cask, USA/9184/B(U) (DOE).

  10. 47 CFR Appendix B to Part 64 - Priority Access Service (PAS) for National Security and Emergency Preparedness (NSEP)

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Security and Emergency Preparedness (NSEP) B Appendix B to Part 64 Telecommunication FEDERAL COMMUNICATIONS.... 64, App. B Appendix B to Part 64—Priority Access Service (PAS) for National Security and Emergency...'s Order to CMRS providers and users to comply with policies and procedures establishing the...

  11. 47 CFR Appendix B to Part 64 - Priority Access Service (PAS) for National Security and Emergency Preparedness (NSEP)

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Security and Emergency Preparedness (NSEP) B Appendix B to Part 64 Telecommunication FEDERAL COMMUNICATIONS.... 64, App. B Appendix B to Part 64—Priority Access Service (PAS) for National Security and Emergency...'s Order to CMRS providers and users to comply with policies and procedures establishing the...

  12. Step-Scan T-Cell Fourier Transform Infrared Photoacoustic Spectroscopy (FTIR-PAS) for Monitoring Environmental Air Pollutants

    NASA Astrophysics Data System (ADS)

    Liu, Lixian; Mandelis, Andreas; Melnikov, Alexander; Michaelian, Kirk; Huan, Huiting; Haisch, Christoph

    2016-07-01

    Air pollutants have adverse effects on the Earth's climate system. There is an urgent need for cost-effective devices capable of recognizing and detecting various ambient pollutants. An FTIR photoacoustic spectroscopy (FTIR-PAS) method based on a commercial FTIR spectrometer developed for air contamination monitoring will be presented. A resonant T-cell was determined to be the most appropriate resonator in view of the low-frequency requirement and space limitations in the sample compartment. Step-scan FTIR-PAS theory for regular cylinder resonator has been described as a reference for prediction of T-cell vibration principles. Both simulated amplitude and phase responses of the T-cell show good agreement with measurement data Carbon dioxide IR absorption spectra were used to demonstrate the capacity of the FTIR-PAS method to detect ambient pollutants. The theoretical detection limit for carbon dioxide was found to be 4 ppmv. A linear response to carbon dioxide concentration was found in the range from 2500 ppmv to 5000 ppmv. The results indicate that it is possible to use step-scan FTIR-PAS with a T-cell as a quantitative method for analysis of ambient contaminants.

  13. The Psychiatric Assessment Schedule for Adult with Developmental Disability (PAS-ADD) Checklist: Reliability and Validity of French Version

    ERIC Educational Resources Information Center

    Gerber, F.; Carminati, G. Galli

    2013-01-01

    Background: The lack of psychometric measures of psychopathology especially in intellectual disabilities (ID) population was addressed by creation of the Psychiatric Assessment Schedule for Adult with Developmental Disability (PAS-ADD-10) in Moss et?al. This schedule is a structured interview designed for professionals in psychopathology. The…

  14. Members of the bHLH-PAS family regulate Shh transcription in forebrain regions of the mouse CNS.

    PubMed

    Epstein, D J; Martinu, L; Michaud, J L; Losos, K M; Fan, C; Joyner, A L

    2000-11-01

    The secreted protein sonic hedgehog (Shh) is required to establish patterns of cellular growth and differentiation within ventral regions of the developing CNS. The expression of Shh in the two tissue sources responsible for this activity, the axial mesoderm and the ventral midline of the neural tube, is regulated along the anteroposterior neuraxis. Separate cis-acting regulatory sequences have been identified which direct Shh expression to distinct regions of the neural tube, supporting the view that multiple genes are involved in activating Shh transcription along the length of the CNS. We show here that the activity of one Shh enhancer, which directs reporter expression to portions of the ventral midbrain and diencephalon, overlaps both temporally and spatially with the expression of Sim2. Sim2 encodes a basic helix-loop-helix (bHLH-PAS) PAS domain containing transcriptional regulator whose Drosophila homolog, single-minded, is a master regulator of ventral midline development. Both vertebrate and invertebrate Sim family members were found sufficient for the activation of the Shh reporter as well as endogenous Shh mRNA. Although Shh expression is maintained in Sim2(-)(/)(-) embryos, it was determined to be absent from the rostral midbrain and caudal diencephalon of embryos carrying a dominant-negative transgene that disrupts the function of bHLH-PAS proteins. Together, these results suggest that bHLH-PAS family members are required for the regulation of Shh transcription within aspects of the ventral midbrain and diencephalon. PMID:11023872

  15. Minor-bodies science with the J-PAS/J-PLUS photometric surveys

    NASA Astrophysics Data System (ADS)

    De Prá, M.; Carvano, J.

    2014-07-01

    The JPAS/J-PLUS is a joint Spanish-Brazilian project that aims to map an area of the sky of 8000 square degrees, in order to measure, with unprecedented accuracy, the redshifts of a large sample of extragalactic objects up magnitude 23. It involves the installation of two telescopes on the Javalambre Mountains, in Spain. The T250 telescope will have an aperture of 250 cm and will be equipped with a camera with a 3 square degree field of view and will use a set of 56 filters (54 narrow band + 2 wide band) covering the 0.3--1.0 micron range, while the T80 telescope (presently on commissioning phase) will have a camera with 2 square degree field of view and will use a set of 12 narrow and intermediate band filters covering the same wavelength range. During its execution, the surveys will also observe a large number of minor Solar System bodies. For those objects, the differential of JPAS/J-PLUS with respect to other large photometric survey is the number and position of the filters used, which will allow a better identification of some taxonomic classes that are not well defined only with SDSS-like filter systems. In particular, the JPAS/J-PLUS data set will allow a robust identification of the 0.7 micron water alteration band that characterize the Ch class in Bus taxonomy. Thus, with JPAS/J-PLUS it will be possible to map the occurrence of water alteration in the present Solar System, which in turn will allow us to put further constraints of the presence of volatiles and of heating processes in the early Solar System. Also, although the survey cadence is not optimized for the discovery of minor bodies, J-PAS/J-PLUS can give an important contribution to the effort of expanding the known population of asteroids and TNOs. We will discuss also the expected efficiency of the J-PLUS/J-PAS surveys in finding minor bodies.

  16. Wintergreen oil: a novel method in Wheatley's trichrome staining technique.

    PubMed

    Salleh, Fatmah Md; Anuar, Tengku Shahrul; Yasin, Azlin Mohd; Moktar, Norhayati

    2012-10-01

    Permanent staining of faecal smears by Wheatley's trichrome technique has been used by many scientists for the detection of parasites in the past and it was found to be highly sensitive. This study was conducted to evaluate the use of Wintergreen oil in comparison with xylene in Wheatley's trichrome staining technique, as the reference technique. In a blind comparison study, 500 collected faecal samples from aboriginal communities were examined. Wintergreen oil was found to be more superior than xylene as a clearing agent in the Wheatley's trichrome staining of polyvinyl alcohol-fixed faecal smears for the identification of intestinal protozoa. Elimination of toxic, carcinogenic, and fire hazards makes Wintergreen oil the preferred choice in routine parasitology examinations. PMID:22986100

  17. Chromosome-specific staining to detect genetic rearrangements

    DOEpatents

    Gray, Joe W.; Pinkel, Daniel; Tkachuk, Douglas; Westbrook, Carol

    2013-04-09

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyzes. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML) and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar but genetically different diseases, and for many prognostic and diagnostic applications.

  18. Analyzing Cell Death by Nuclear Staining with Hoechst 33342.

    PubMed

    Crowley, Lisa C; Marfell, Brooke J; Waterhouse, Nigel J

    2016-01-01

    The nuclei of healthy cells are generally spherical, and the DNA is evenly distributed. During apoptosis the DNA becomes condensed, but this process does not occur during necrosis. Nuclear condensation can therefore be used to distinguish apoptotic cells from healthy cells or necrotic cells. Dyes that bind to DNA, such as Hoechst 33342 or 4',6-diamidino-2-phenylindole (DAPI), can be used to observe nuclear condensation. These dyes fluoresce at 461 nm when excited by ultraviolet light and can therefore be visualized using conventional fluorescent microscopes equipped with light sources that emit light at ∼350 nm and filter sets that permit the transmission of light at ∼460 nm. This protocol describes staining and visualization of cells stained with Hoechst 33342, but it can be adapted for staining with DAPI or other dyes. PMID:27587774

  19. A staining protocol for identifying secondary compounds in Myrtaceae1

    PubMed Central

    Retamales, Hernan A.; Scharaschkin, Tanya

    2014-01-01

    • Premise of the study: Here we propose a staining protocol using toluidine blue (TBO) and ruthenium red to reliably identify secondary compounds in the leaves of some species of Myrtaceae. • Methods and Results: Leaves of 10 species representing 10 different genera of Myrtaceae were processed and stained using five different combinations of ruthenium red and TBO. Optimal staining conditions were determined as 1 min of ruthenium red (0.05% aqueous) and 45 s of TBO (0.1% aqueous). Secondary compounds clearly identified under this treatment include mucilage in the mesophyll, polyphenols in the cuticle, lignin in fibers and xylem, tannins and carboxylated polysaccharides in the epidermis, and pectic substances in the primary cell walls. • Conclusions: Potential applications of this protocol include systematic, phytochemical, and ecological investigations in Myrtaceae. It might be applicable to other plant families rich in secondary compounds and could be used as a preliminary screening method for extraction of these elements. PMID:25309840

  20. Pasteurella pestis detection in fleas by fluorescent antibody staining*

    PubMed Central

    Hudson, Bruce W.; Kartman, Leo; Prince, Frank M.

    1966-01-01

    In an effort to develop a method for the rapid field identification of plague-infected fleas, the authors have studied the feasibility of direct fluorescent antibody staining of the midgut contents of fleas fed on mice infected with Pasteurella pestis. Fluorescent antibodies prepared from antisera derived from rabbits inoculated with the water-soluble P. pestis fraction 1b antigen, the somatic antigen of heat-killed P. pestis (Bryans strain), and live avirulent (strain A1122) or virulent (Yreka strain) plague vaccines were used used in this study. This direct staining method proved to be impracticable, but encouraging results were obtained by fluorescent antibody staining of broth cultures of macerates of infected fleas after 24-48 hours' incubation. The broth enrichment technique has not yet been evaluated in the field, but it is expected to be of value since it is relatively simple to perform and requires only material that can easily be transported to remote areas. PMID:5328902

  1. Stain-free histopathology by programmable supercontinuum pulses

    NASA Astrophysics Data System (ADS)

    Tu, Haohua; Liu, Yuan; Turchinovich, Dmitry; Marjanovic, Marina; Lyngsø, Jens K.; Lægsgaard, Jesper; Chaney, Eric J.; Zhao, Youbo; You, Sixian; Wilson, William L.; Xu, Bingwei; Dantus, Marcos; Boppart, Stephen A.

    2016-08-01

    The preparation, staining, visualization and interpretation of histological images of tissue is well accepted as the gold standard process for the diagnosis of disease. These methods have a long history of development, and are used ubiquitously in pathology, despite being highly time- and labour-intensive. Here, we introduce a unique optical imaging platform and methodology for label-free multimodal multiphoton microscopy that uses a novel photonic-crystal fibre source to generate tailored chemical contrast based on programmable supercontinuum pulses. We demonstrate the collection of optical signatures of the tumour microenvironment, including evidence of mesoscopic biological organization, tumour cell migration and (lymph-) angiogenesis collected directly from fresh ex vivo mammary tissue. Acquisition of these optical signatures and other cellular or extracellular features, which are largely absent from histologically processed and stained tissue, combined with an adaptable platform for optical alignment-free programmable-contrast imaging, offers the potential to translate stain-free molecular histopathology into routine clinical use.

  2. An accurate cluster selection function for the J-PAS narrow-band wide-field survey

    NASA Astrophysics Data System (ADS)

    Ascaso, B.; Benítez, N.; Dupke, R.; Cypriano, E.; Lima-Neto, G.; López-Sanjuan, C.; Varela, J.; Alcaniz, J. S.; Broadhurst, T.; Cenarro, A. J.; Devi, N. Chandrachani; Díaz-García, L. A.; Fernandes, C. A. C.; Hernández-Monteagudo, C.; Mei, S.; Mendes de Oliveira, C.; Molino, A.; Oteo, I.; Schoenell, W.; Sodré, L.; Viironen, K.; Marín-Franch, A.

    2016-03-01

    The impending Javalambre Physics of the accelerating Universe Astrophysical Survey (J-PAS) will be the first wide-field survey of ≳ 8500 deg2 to reach the `stage IV' category. Because of the redshift resolution afforded by 54 narrow-band filters, J-PAS is particularly suitable for cluster detection in the range z<1. The photometric redshift dispersion is estimated to be only ˜0.003 with few outliers ≲4 per cent for galaxies brighter than i ˜ 23 AB, because of the sensitivity of narrow band imaging to absorption and emission lines. Here, we evaluate the cluster selection function for J-PAS using N-body+semi-analytical realistic mock catalogues. We optimally detect clusters from this simulation with the Bayesian Cluster Finder, and we assess the completeness and purity of cluster detection against the mock data. The minimum halo mass threshold we find for detections of galaxy clusters and groups with both >80 per cent completeness and purity is Mh ˜ 5 × 1013 M⊙ up to z ˜ 0.7. We also model the optical observable, M^{*}_CL-halo mass relation, finding a non-evolution with redshift and main scatter of σ _{M^{*}_CL | M_h}˜ 0.14 dex down to a factor 2 lower in mass than other planned broad-band stage IV surveys, at least. For the Mh ˜ 1 × 1014 M⊙ Planck mass limit, J-PAS will arrive up to z ˜ 0.85 with a σ _{M^{*}_CL | M_h}˜ 0.12 dex. Therefore, J-PAS will provide the largest sample of clusters and groups up to z ˜ 0.8 with a mass calibration accuracy comparable to X-ray data.

  3. Stain Specific Standardization of Whole-Slide Histopathological Images.

    PubMed

    Bejnordi, Babak Ehteshami; Litjens, Geert; Timofeeva, Nadya; Otte-Höller, Irene; Homeyer, André; Karssemeijer, Nico; van der Laak, Jeroen A W M

    2016-02-01

    Variations in the color and intensity of hematoxylin and eosin (H&E) stained histological slides can potentially hamper the effectiveness of quantitative image analysis. This paper presents a fully automated algorithm for standardization of whole-slide histopathological images to reduce the effect of these variations. The proposed algorithm, called whole-slide image color standardizer (WSICS), utilizes color and spatial information to classify the image pixels into different stain components. The chromatic and density distributions for each of the stain components in the hue-saturation-density color model are aligned to match the corresponding distributions from a template whole-slide image (WSI). The performance of the WSICS algorithm was evaluated on two datasets. The first originated from 125 H&E stained WSIs of lymph nodes, sampled from 3 patients, and stained in 5 different laboratories on different days of the week. The second comprised 30 H&E stained WSIs of rat liver sections. The result of qualitative and quantitative evaluations using the first dataset demonstrate that the WSICS algorithm outperforms competing methods in terms of achieving color constancy. The WSICS algorithm consistently yields the smallest standard deviation and coefficient of variation of the normalized median intensity measure. Using the second dataset, we evaluated the impact of our algorithm on the performance of an already published necrosis quantification system. The performance of this system was significantly improved by utilizing the WSICS algorithm. The results of the empirical evaluations collectively demonstrate the potential contribution of the proposed standardization algorithm to improved diagnostic accuracy and consistency in computer-aided diagnosis for histopathology data. PMID:26353368

  4. Modeling of alkane emissions from a wood stain

    SciTech Connect

    Chang, J.C.S.; Guo, Z.

    1993-01-01

    The article discusses full-scale residential house tests to evaluate the effects of organic emissions from a wood finishing product--wood stain--on indoor air quality (IAQ). The test house concentrations of three alkane species, nonane, decane, and undecane, were measured as a function of time after the application of the wood stain. It was found that the test house concentrations can be simulated by an integrated IAQ model which takes into consideration source, sink, and ventilation effects. The alkane emissions were controlled by an evaporation-like process.

  5. Recurrent oral pyogenic granuloma in port-wine stain.

    PubMed

    da Silva, Alessandra Dutra; Silva, Carolina Amália Barcellos; de Camargo Moraes, Paulo; Thomaz, Luiz Alexandre; Furuse, Cristiane; de Araújo, Vera Cavalcanti

    2011-11-01

    Pyogenic granuloma (PG) is a benign inflammatory lesion, nonneoplastic in nature, which occurs in the oral cavity and skin. This lesion arises in response to various stimuli such as low-grade local irritations, traumatic injury, or hormonal factors. Recently, in some cases, the occurrence of recurrent PGs in skin associated with vascular lesions, such as port-wine stains, has been described. It has been postulated that this association is promoted by arteriovenous anastomoses in the vascular lesions, leading to the development of PG. The authors discuss 2 cases of recurrent PG in patients with a port-wine stain, and the treatment options adopted. PMID:22134277

  6. Detection of Hydrogen Peroxide by DAB Staining in Arabidopsis Leaves

    PubMed Central

    Daudi, Arsalan; O’Brien, Jose A.

    2016-01-01

    In this protocol, the in situ detection of hydrogen peroxide (one of several reactive oxygen species) is described in mature Arabidopsis rosette leaves by staining with 3,3′-diaminobenzidine (DAB) using an adaptation of previous methods (Thordal-Christensen et al., 1997; Bindschedler et al., 2006; Daudi et al., 2012). DAB is oxidized by hydrogen peroxide in the presence of some haem-containing proteins, such as peroxidases, to generate a dark brown precipitate. This precipitate is exploited as a stain to detect the presence and distribution of hydrogen peroxide in plant cells. The protocol can be modified slightly to detect hydrogen peroxide in different types of plant tissue.

  7. Staining Fission Yeast Filamentous Actin with Fluorescent Phalloidin Conjugates.

    PubMed

    Hagan, Iain M

    2016-01-01

    The Schizosaccharomyces pombe filamentous (F)-actin cytoskeleton drives cell growth, morphogenesis, endocytosis, and cytokinesis. The protocol described here reveals the distribution of F-actin in fixed cells through the use of fluorescently conjugated phalloidin. Simultaneous staining of cell wall landmarks (with calcofluor) and chromatin (with 4',6-diamidino-2-phenylindole, or DAPI) makes this rapid staining procedure highly effective for staging cell cycle progression, monitoring morphogenetic abnormalities, and assessing the impact of environmental and genetic changes on the integrity of the F-actin cytoskeleton. PMID:27250943

  8. Determination of firing distance using the rhodizonate staining technique.

    PubMed

    Marty, W; Sigrist, T; Wyler, D

    2002-02-01

    The histological staining technique using rhodizonate is also effective for the determination of the firing distance by examining the distribution and intensity of the staining reaction. The differentiation between absolute close-range shots and long-range shots is generally possible without any doubt. The method is not recommended for routine examinations but it is very useful for cases lacking the possibility to investigate smoke and powder deposits in a criminalistic manner, i.e. surgical skin biopsies of hospitalised victims and skin highly altered by the effects of fire, water or by post-mortem decomposition. PMID:11924700

  9. Chemical aspects of santalin as a histological stain.

    PubMed

    Banerjee, A; Mukherjee, A K

    1981-03-01

    Recent research on the chemical nature of the red dyes isolated from Pterocarpus santalinus and certain West African plants, viz., Baphia nitida, Pterocarpus osun and Pterocarpus soyauxii, have been reviewed. P. santalinus contains santalins A, B and C, but no santarubin. Santalins and santarubins have been found in P. osun, P. soyauxii and B. nitida. The structural formulae of the santalins are presented and their differences from santarubins indicated. Santalins A and B have some similarities in structure with hematein. This is probably responsible for their staining properties; the possible mechanism of staining is discussed. PMID:6166100

  10. The Neuronal PAS Domain Protein 4 (Npas4) Is Required for New and Reactivated Fear Memories

    PubMed Central

    Nguyen, Tam; DiLeone, Ralph J.; Schafe, Glenn E.

    2011-01-01

    The Neuronal PAS domain protein 4 (Npas4) is a neuronal activity-dependent immediate early gene that has recently been identified as a transcription factor which regulates the transcription of genes that control inhibitory synapse development and synaptic plasticity. The role Npas4 in learning and memory, however, is currently unknown. Here, we systematically examine the role of Npas4 in auditory Pavlovian fear conditioning, an amygdala-dependent form of emotional learning. In our first series of experiments, we show that Npas4 mRNA and protein are regulated in the rat lateral nucleus of the amygdala (LA) in a learning-dependent manner. Further, knockdown of Npas4 protein in the LA via adeno-associated viral (AAV) mediated gene delivery of RNAi was observed to impair fear memory formation, while innate fear and the expression of fear memory were not affected. In our second series of experiments, we show that Npas4 protein is regulated in the LA by retrieval of an auditory fear memory and that knockdown of Npas4 in the LA impairs retention of a reactivated, but not a non-reactivated, fear memory. Collectively, our findings provide the first comprehensive look at the functional role of Npas4 in learning and memory. PMID:21887312

  11. iPAS: AES Flight System Technology Maturation for Human Spaceflight

    NASA Technical Reports Server (NTRS)

    Othon, William L.

    2014-01-01

    In order to realize the vision of expanding human presence in space, NASA will develop new technologies that can enable future crewed spacecraft to go far beyond Earth orbit. These technologies must be matured to the point that future project managers can accept the risk of incorporating them safely and effectively within integrated spacecraft systems, to satisfy very challenging mission requirements. The technologies must also be applied and managed within an operational context that includes both on-board crew and mission support on Earth. The Advanced Exploration Systems (AES) Program is one part of the NASA strategy to identify and develop key capabilities for human spaceflight, and mature them for future use. To support this initiative, the Integrated Power Avionics and Software (iPAS) environment has been developed that allows engineers, crew, and flight operators to mature promising technologies into applicable capabilities, and to assess the value of these capabilities within a space mission context. This paper describes the development of the integration environment to support technology maturation and risk reduction, and offers examples of technology and mission demonstrations executed to date.

  12. The development of the digestive tract in larval European catfish (Silurus glanis L.).

    PubMed

    Kozarić, Z; Kuzir, S; Petrinec, Z; Gjurcević, E; Bozić, M

    2008-04-01

    The European catfish, Silurus glanis L., has become an important aqua cultural fish in Croatia, and it is cultivated extensively in ponds in polyculture with carps. The development of the digestive tract in S. glanis was studied with the aim of improving intensive fish production. Research was carried out on S. glanis larval stadium from 1- to 19-day post-hatching (DPH). The main histological methods used were: haematoxylin and eosin staining, periodic acid Schiff staining (PAS), Alcian blue (AB) and toluidin blue staining (TB). A yolk sac was present during the first 5 days (1-5-DPH). During the initial 3-DPH period, there was no trace PAS and AB activity in the digestive tract. Differentiation of the digestive tract began at 3- to 5-DPH. The oesophagus was positive for AB at 5-DPH, PAS and TB after 7-DPH. Differentiation of enterocytes began at 5-DPG and the intestines were complete at 11-DPH. Development of liver and pancreas was also studied. The analysis of data obtained in this study suggests that after 5-DPH catfish larvae have morphologically completed digestive tracts. PMID:18333856

  13. Routine histologic examination for the diagnosis of onychomycosis: an evaluation of sensitivity and specificity.

    PubMed

    Machler, B C; Kirsner, R S; Elgart, G W

    1998-04-01

    Clinical differentiation of dermatophyte infection from dystrophic changes due to psoriasis may be challenging. Typically, potassium hydroxide (KOH) preparations, fungal culture, and occasionally, nail unit biopsy specimens are utilized to help differentiate between the two. These tests are often time-consuming and may yield false-negative results. Increasing regulation of the office laboratory has caused some physicians to forgo this testing, which was previously routine. We investigated the utility of routine histologic examination of nail clippings in differentiating onychomycosis from psoriatic onychodystrophy. Twenty-three distal nail clipping specimens (twelve specimens from patients with onychodystrophy of unknown cause and eleven control specimens from nails with known cause) were evaluated by routine histology and periodic acid-Schiff (PAS) staining. Of the dystrophic cases, four were demonstrated to be onychomycosis by the presence of hyphae on histologic evaluation and by culture, whereas only three of these cases yielded positive results on KOH examination. Eight cases of onychodystrophy were due to psoriasis. Yeast forms were detected on one case of psoriatic onychodystrophy that demonstrated yeast growth on culture. In our study, routine histologic examination with PAS staining was equal to culture and superior to KOH preparation in leading to the correct diagnosis of dermatophyte infection. In addition, the diagnosis of psoriasis of the nail plate was detected accurately by routine histologic examination. Routine histologic examination with PAS staining is a rapid, simple, and reliable test in the evaluation of onychodystrophy. PMID:9564595

  14. A two-color acid-free cartilage and bone stain for zebrafish larvae.

    PubMed

    Walker, M B; Kimmel, C B

    2007-02-01

    Traditionally, cartilage is stained by alcian blue using acidic conditions to differentiate tissue staining. The acidic conditions are problematic when one wishes to stain the same specimen for mineralized bone with alizarin red, because acid demineralizes bone, which negatively affects bone staining. We have developed an acid-free method to stain cartilage and bone simultaneously in zebrafish larvae. This method has the additional advantage that PCR genotyping of stained specimens is possible. PMID:17510811

  15. Evaluation of a fluorescent lectin-based staining technique for some acidophilic mining bacteria

    SciTech Connect

    Fife, D.J.; Bruhn, D.F.; Miller, K.S.; Stoner, D.L.

    2000-05-01

    A fluorescence-labeled wheat germ agglutinin staining technique was modified and found to be effective for staining gram-positive, acidophilic mining bacteria. Bacteria identified by others as being gram positive through 16S rRNA sequence analyses, yet clustering near the divergence of that group, stained weakly. Gram-negative bacteria did not stain. Background staining of environmental samples was negligible, and pyrite and soil particles in the samples did not interfere with the staining procedure.

  16. Candida and calcofluor white: Study in precancer and cancer

    PubMed Central

    Kumar, Rashmi Santosh; Ganvir, SM; Hazarey, VK

    2009-01-01

    Background: The interest in oral candidosis has waxed and waned from the period of Hippocrates. The acquired immune deficiency syndrome (AIDS) epidemic has certainly bolstered these figures on oral candidosis, with diabetes and oral cancer being no exception. A need for rapid detection of Candida is made possible by the use of Calcofluor - White (CFW) stain when examined under a fluorescence microscope. The present study was aimed at assessing the efficacy of CFW is compared to Gram stain and periodic acid Schiff (PAS) in detection of Candida in oral precancer and cancer. Materials and Methods: The study group consisted of patients with precancer (n=45), cancer (n=45), and control group (n=45). Presence of Candida was confirmed by culture inoculation along with a germ tube and carbohydrate fermentation test. The cytopathological smears were analyzed by papanicolaou - CFW and Gram staining, whereas, tissue sections were stained by PAS and CFW staining. Results: Candida albicans was the predominant species identified. A highly significant association of Candida was seen more often in cancer than in precancer. Both in cytology and histopathology Candida detection by CFW was higher. In precancer it was 48.88% in smears and 40% in tissue sections, whereas, in cancer 60% in smears and 55.55% in histopathology. Conclusion: Among the various diagnostic tools used in the present study, the use of CFW is seen to be a simple, effective, rapid, and reliable method, both in cytopathology and histopathology. PMID:21886989

  17. MODELING OF ALKANE EMISSIONS FROM A WOOD STAIN

    EPA Science Inventory

    The article discusses full-scale residential house tests to evaluate the effects of organic emissions from a wood finishing product--wood stain--on indoor air quality (IAQ). The test house concentrations of three alkane species, nonane, decane, and undecane, were measured as a fu...

  18. Image analysis of dye stained patterns in soils

    NASA Astrophysics Data System (ADS)

    Bogner, Christina; Trancón y Widemann, Baltasar; Lange, Holger

    2013-04-01

    Quality of surface water and groundwater is directly affected by flow processes in the unsaturated zone. In general, it is difficult to measure or model water flow. Indeed, parametrization of hydrological models is problematic and often no unique solution exists. To visualise flow patterns in soils directly dye tracer studies can be done. These experiments provide images of stained soil profiles and their evaluation demands knowledge in hydrology as well as in image analysis and statistics. First, these photographs are converted to binary images classifying the pixels in dye stained and non-stained ones. Then, some feature extraction is necessary to discern relevant hydrological information. In our study we propose to use several index functions to extract different (ideally complementary) features. We associate each image row with a feature vector (i.e. a certain number of image function values) and use these features to cluster the image rows to identify similar image areas. Because images of stained profiles might have different reasonable clusterings, we calculate multiple consensus clusterings. An expert can explore these different solutions and base his/her interpretation of predominant flow mechanisms on quantitative (objective) criteria. The complete workflow from reading-in binary images to final clusterings has been implemented in the free R system, a language and environment for statistical computing. The calculation of image indices is part of our own package Indigo, manipulation of binary images, clustering and visualization of results are done using either build-in facilities in R, additional R packages or the LATEX system.

  19. [Pnemocystis jiroveci pneumonia: Comparison between conventional PCR and staining techniques].

    PubMed

    Kaouech, E; Kallel, K; Anane, S; Belhadj, S; Abdellatif, S; Mnif, K; Ben Othmane, T; Ben Lakhal, S; Kilani, B; Ben Châabane, T; Chaker, E

    2009-07-01

    Diagnosis of pneumocystis pneumonia is usually based on clinical features and X-rays photography and confirmed in the laboratory by visualisation of Pneumocystis organisms in stained preparations of respiratory specimens using several techniques (Gomori-Grocott, May-Grünwald Giemsa, bleu de toluidine O). Actually, PCR has considerably increased sensitivity of detection of Pneumocystis. The aim of this study is to compare conventional PCR results to those of staining techniques (Gomori-Grocott, May-Grünwald Giemsa) in addition to the X-ray and clinical findings in order to evaluate the contribution of each method. Sixty-four respiratory specimens were collected from 54 immuno-compromised patients with clinical symptoms of pulmonary infection. We diagnosed pneumocystis pneumonia in 16 patients according to staining techniques and/or typical clinical and radiological findings and/or response to treatment. Of the 15 patients, 14 were positive by PCR and only five were positive by direct examination, yielding a sensitivity and specificity of 93.3 and 87.1% for PCR and 33.3 and 100% for staining techniques. Conventional PCR provides a sensitive and objective method for the detection Pneumocystis jiroveci from less invasive sample. PMID:19038508

  20. ANEUPLOIDY TEST DEVELOPMENT: KINETOCHORE STAINING IN MAMMALIAN SYSTEMS

    EPA Science Inventory

    The purpose of the project was to determine the feasibility of using human-derived antibodies against the chromosomal kinetochore region coupled with immunofluorescence staining as a method for evaluating the induction of aneuploidy in mammalian cells in vitro and in vivo. The te...

  1. Lipophilic dye staining of Cryptococcus neoformans extracellular vesicles and capsule.

    PubMed

    Nicola, André Moraes; Frases, Susana; Casadevall, Arturo

    2009-09-01

    Cryptococcus neoformans is an encapsulated yeast that causes systemic mycosis in immunosuppressed individuals. Recent studies have determined that this fungus produces vesicles that are released to the extracellular environment both in vivo and in vitro. These vesicles contain assorted cargo that includes several molecules associated with virulence and implicated in host-pathogen interactions, such as capsular polysaccharides, laccase, urease, and other proteins. To date, visualization of extracellular vesicles has relied on transmission electron microscopy, a time-consuming technique. In this work we report the use of fluorescent membrane tracers to stain lipophilic structures in cryptococcal culture supernatants and capsules. Two dialkylcarbocyanine probes with different spectral characteristics were used to visualize purified vesicles by fluorescence microscopy and flow cytometry. Dual staining of vesicles with dialkylcarbocyanine and RNA-selective nucleic acid dyes suggested that a fraction of the vesicle population carried RNA. Use of these dyes to stain whole cells, however, was hampered by their possible direct binding to capsular polysaccharide. A fluorescent phospholipid was used as additional membrane tracer to stain whole cells, revealing punctate structures on the edge of the capsule which are consistent with vesicular trafficking. Lipophilic dyes provide new tools for the study of fungal extracellular vesicles and their content. The finding of hydrophobic regions in the capsule of C. neoformans adds to the growing evidence for a structurally complex structure composed of polysaccharide and nonpolysaccharide components. PMID:19465562

  2. 31. Interior detail view of arched, steelframed, stained glass windows ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    31. Interior detail view of arched, steel-framed, stained glass windows at the landing of the south stairs in main lobby, view looking south from second floor lobby - University of Oregon Museum of Art, 1470 Johnson Lane, Eugene, Lane County, OR

  3. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND...

  4. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND STANDARD...

  5. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND STANDARD...

  6. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND...

  7. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND...

  8. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND...

  9. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Strict Middling Yellow Stained Color. 28.441 Section 28.441 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND...

  10. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND STANDARD...

  11. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND STANDARD...

  12. 7 CFR 28.442 - Middling Yellow Stained Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE COMMODITY STANDARDS AND STANDARD...

  13. Analysis of Oxiclean: An Interesting Comparison of Percarbonate Stain Removers

    ERIC Educational Resources Information Center

    Bracken, Jeffrey D.; Tietz, David

    2005-01-01

    The study focuses on percarbonate-based stain removers since the percarbonate can be heated to produce additional sodium carbonate. An experiment that provides general chemistry students an opportunity to apply their knowledge of basic stoichiometry to solve a relevant, real-world problem is described.

  14. A conservative approach to esthetically treat stained arrested caries lesions.

    PubMed

    Al-Angari, Sarah S; Hara, Anderson T

    2016-01-01

    Esthetic treatment of stained arrested caries lesions (ACLs) has mostly been done using invasive restorative techniques. The aim of this paper was to propose and report the efficacy of a conservative approach based on dental bleaching to esthetically treat these lesions, both experimentally (extracted teeth) and clinically. In a laboratory experiment, ten extracted human teeth with stained ACLs in either pit and fissure or smooth surface were selected and treated with 15% carbamide peroxide gel, 4 h per day, for a total of 6 days. The second part of the paper reports a clinical case of pit and fissure-stained ACLs in four posterior teeth, which were treated with 40% hydrogen peroxide in-office bleaching. Digital photographs were taken in both parts to document the efficacy of the treatment. The lesions showed noticeable increase in color lightness indicating the efficacy and suitability of the proposed approach. By using the conservative clinical technique presented, the esthetics of most stained ACLs could be improved, eliminating the need for invasive restorative treatments. PMID:27092359

  15. 5. Downstream elevation, view to southeast. Dark stains on side ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    5. Downstream elevation, view to southeast. Dark stains on side of main girder are from deck drain scuppers, marking deck level within the girders. Compare this view and CA-126-7 to CA-126-19 for indication of severity of siltation of Salt River channel has silted. - Salt River Bridge, Spanning Salt River at Dillon Road, Ferndale, Humboldt County, CA

  16. MODELING OF ALKANE EMISSIONS FROM A WOOD STAIN

    EPA Science Inventory

    The article discusses full-scale residential house tests to evaluate the effects of organic emissions from a wood finishing product--wood stain--on indoor air quality (IAQ). he test house concentrations of three alkane species, nonane, decane, and undecane, were measured as a fun...

  17. Specific immunofluorescence staining of Treponema pallidum in smears and tissues.

    PubMed

    Ito, F; Hunter, E F; George, R W; Swisher, B L; Larsen, S A

    1991-03-01

    To date, tissue sections prepared from Formalin-fixed tissues have not been successfully stained with Treponema pallidum subspecies-specific antibody in a direct fluorescent-antibody assay. While current methods stain T. pallidum, they do not distinguish T. pallidum from other spirochetes such as Borrelia burgdorferi (E. F. Hunter, P. W. Greer, B. L. Swisher, A. R. Simons, C. E. Farshy, J. A. Crawford, and K. R. Sulzer, Arch. Pathol. Lab. Med. 108:878-880, 1984). Because trypsin pretreatment of tissue sections has enhanced other immunofluorescent-antibody (IFA) applications, we compared the use of the trypsin digestion method with the current 1% ammonium hydroxide (NH4OH) method as a means to obtain specific staining of T. pallidum in tissues by both direct and indirect IFA techniques. Pretreated T. pallidum-infected tissues sections from rabbits, hamsters, and humans were quantitatively examined with the direct fluorescent-antibody-T. pallidum test conjugate absorbed with Treponema phagedenis, the Reiter treponeme. For indirect staining, a serum specimen from a patients with syphilis absorbed by affinity chromatography with T. phagedenis was used as the primary reagent, and a fluorescein isothiocyanate-labeled rabbit anti-human globulin was used as the secondary reagent. Serum specificity was established first by examining antigen smears of T. pallidum subsp. pallidum, T. pallidum subsp. pertenue, B. burgdorferi, T. phagedenis, and Treponema denticola MRB and then by examining tissues infected with these pathogens plus those infected with four Leptospira serovars. When we stained tissue using the direct IFA method that is currently a standard method for the examination of chancre smears, we found it to be unsuitable for use with tissue. Trypsin digestion did not offer an improvement over the NH4OH pretreatment method in the specific identification of T. pallidum by direct IFA. However, specific identification of T. pallidum in tissue sections was obtained by the

  18. Cloche is a bHLH-PAS transcription factor that drives haemato-vascular specification.

    PubMed

    Reischauer, Sven; Stone, Oliver A; Villasenor, Alethia; Chi, Neil; Jin, Suk-Won; Martin, Marcel; Lee, Miler T; Fukuda, Nana; Marass, Michele; Witty, Alec; Fiddes, Ian; Kuo, Taiyi; Chung, Won-Suk; Salek, Sherveen; Lerrigo, Robert; Alsiö, Jessica; Luo, Shujun; Tworus, Dominika; Augustine, Sruthy M; Mucenieks, Sophie; Nystedt, Björn; Giraldez, Antonio J; Schroth, Gary P; Andersson, Olov; Stainier, Didier Y R

    2016-07-14

    Vascular and haematopoietic cells organize into specialized tissues during early embryogenesis to supply essential nutrients to all organs and thus play critical roles in development and disease. At the top of the haemato-vascular specification cascade lies cloche, a gene that when mutated in zebrafish leads to the striking phenotype of loss of most endothelial and haematopoietic cells and a significant increase in cardiomyocyte numbers. Although this mutant has been analysed extensively to investigate mesoderm diversification and differentiation and continues to be broadly used as a unique avascular model, the isolation of the cloche gene has been challenging due to its telomeric location. Here we used a deletion allele of cloche to identify several new cloche candidate genes within this genomic region, and systematically genome-edited each candidate. Through this comprehensive interrogation, we succeeded in isolating the cloche gene and discovered that it encodes a PAS-domain-containing bHLH transcription factor, and that it is expressed in a highly specific spatiotemporal pattern starting during late gastrulation. Gain-of-function experiments show that it can potently induce endothelial gene expression. Epistasis experiments reveal that it functions upstream of etv2 and tal1, the earliest expressed endothelial and haematopoietic transcription factor genes identified to date. A mammalian cloche orthologue can also rescue blood vessel formation in zebrafish cloche mutants, indicating a highly conserved role in vertebrate vasculogenesis and haematopoiesis. The identification of this master regulator of endothelial and haematopoietic fate enhances our understanding of early mesoderm diversification and may lead to improved protocols for the generation of endothelial and haematopoietic cells in vivo and in vitro. PMID:27411634

  19. The bHLH/PAS transcription factor singleminded 2s promotes mammary gland lactogenic differentiation.

    PubMed

    Wellberg, Elizabeth; Metz, Richard P; Parker, Caitlin; Porter, Weston W

    2010-03-01

    We have previously demonstrated that the bHLH/PAS transcription factor, singleminded 2s (Sim2s), is required for proper mammary ductal morphogenesis and luminal epithelial differentiation. Furthermore, loss of Sim2s in breast cancer cells resulted in downregulation of epithelial markers and acquisition of a basal-like phenotype. The objective of this study was to further define the role of Sim2s in mammary differentiation. We found that Sim2s is developmentally regulated throughout mammary gland development with highest expression during lactation. Mammary glands from nulliparous mice expressing Sim2s driven by the mouse mammary tumor virus (MMTV) long terminal repeat (LTR) promoter were morphologically indistinguishable from wild-type mice but displayed hallmarks of precocious lactogenic differentiation. These included elevated expression of the milk protein genes Wap and Csn2, and apical localization of the lactation marker Npt2b. Consistent with the in vivo results, Sim2s enhanced prolactin-mediated Csn2 expression in HC11 and CIT3 mouse mammary epithelial cells, and downregulation of Sim2s by shRNA in HC11 cells inhibited Csn2 expression. Chromatin immunoprecipitation (ChIP) analyses of the Csn2 gene found that Sim2s associates with the Csn2 promoter and re-ChIP experiments showed that Sim2s interacted with the RNA II polymerase (RNAPII) complex. Together, these data demonstrate, for the first time, that Sim2s is required for establishing and maintaining mammary gland differentiation. PMID:20150276

  20. Stain and dye stability over a 30-year period: a comparison of certified dye powders by the Biological Stain Commission.

    PubMed

    Penney, D P; Frank, M; Fagan, C; Willis, C

    2009-02-01

    The Biological Stain Commission (BSC) Assay Laboratory has received numerous inquiries during the past several years regarding the long-term stability of stain and dye powders, particularly since packaging requirements call for expiration dates on reagents. We have conducted a study to examine the long-term stability of selected dye powders. We used the standard procedures of the BSC for testing biological stains for certification to give an indication of the long-term chemical stability as well as staining performance of the dye powders. An earlier study by Emmel and Stotz examined the stability of various dye powders after a five-year storage period. The present study is a follow-up project covering the same dyes after storage for 30 years. The dye samples chosen for the study are the same samples used in the five-year storage period study and give comparative results for all three time periods. The results of this study affirm the generally held speculation that dye powders are stable for many years and thus have a substantial shelf-life. PMID:19096966

  1. Toxic pyrrolizidinalkaloids as undesired contaminants in food and feed: degradation of the PAs from Senecio jacobaea in silage.

    PubMed

    Becerra-Jiminez, J; Kuschak, M; Roeder, E; Wiedenfeld, H

    2013-07-01

    Pyrrolizidine alkaloids (PAs) can show a hazardous potential for men and animals. They can act as cancerogenic, mutagenic, teratogenic and fetotoxic agents. One pathway of a human intoxication is its occurence as contaminants in food and feed. Here, the contamination of cereals already led to severe and fatal intoxication episodes. Besides this, milk is of special concern as it is the main food for children which show a very high susceptibility for a PA intoxication. Milk can contain PAs in case the milk producing animals have access to contaminated feed. In this context it is of special interest whether the PA content of contaminated silage remains stable during the ensiling procedure or show a more or less high level of decomposition. We could show that ensiling will not lead to PA-free silage. PMID:23923650

  2. Vasculogenic mimicry has no prognostic significance in pT3 and pT4 cutaneous melanoma.

    PubMed

    Massi, Daniela; Franchi, Alessandro; Paglierani, Milena; Ketabchi, Sheyda; Borgognoni, Lorenzo; Reali, Umberto Maria; Santucci, Marco

    2004-04-01

    The concept of vasculogenic mimicry has been introduced to define periodic acid-Schiff (PAS)-positive channels and loops lined by tumor cells, instead of endothelium, able to contribute to microcirculation in uveal melanomas. Previous studies have shown that the PAS-positive patterns are associated with a poor prognosis in uveal melanoma. The aim of the current study was to investigate whether vasculogenic mimicry has a prognostic impact in pT3 and pT4 cutaneous melanoma. Fifteen patients with pT3 and pT4 cutaneous melanoma who did not experience progression after 10 years of follow-up and 30 matched controls who underwent progression were selected. Tumor sections were stained with PAS reaction, omitting the nuclear counterstaining. For immunohistochemistry, sections were stained with CD31, CD105 (endoglin), and laminin. Differences in the distribution of the PAS-positive patterns and a series of clinicopathological variables were evaluated by the Pearson chi(2) and Mann-Whitney U tests. We observed PAS-positive linear sheets, arcs, elliptical loops, and networks encircling roundish to oval aggregates of melanoma cells. The overall distribution of the PAS-positive patterns did not match with the blood microvessels' architecture as detected by immunohistochemical analysis. No statistically significant differences in the distribution of PAS-positive patterns were found between cases and controls. The presence of a parallel pattern correlated significantly with thickness (P = 0.04), whereas an inverse correlation was found with vessel area (P = 0.05). In conclusion, our results suggest that there is a mismatch between vasculogenic mimicry and tumor angiogenesis and do not support any prognostic role of vasculogenic mimicry in thick cutaneous melanoma. PMID:15116332

  3. Cross contamination of cytological smears, with automated staining machines and bulk manual staining procedures. With a specific study of the problems of the Cytotek and the Shandon Elliott staining machines.

    PubMed

    Husain, O A; Grainger, J M; Sims, J

    1978-01-01

    Further development of an individual staining machine is to be strongly encouraged but meanwhile, using bulk stainers, frequent changing of wash fluids and staining solutions, particularly leading up to and following the haematoxylin pot, is essential to reduce the risk of cross contamination. Certain smears, such as from semen or from serous fluids where malignancy is suspected or known, must be stained on separate racks. In some laboratories it is the rule not to stain semen or serous fluids in bulk staining machines at all and this may have to become the rule everywhere until we are provided with safe individual slide stainers. PMID:75214

  4. Determination of drug content in semisolid formulations by non-invasive spectroscopic methods: FTIR - ATR, - PAS, - Raman and PDS

    NASA Astrophysics Data System (ADS)

    Gotter, B.; Faubel, W.; Heißler, St.; Hein, J.; Neubert, R. H. H.

    2010-03-01

    This study elucidates the potential use of photothermal deflection spectroscopy (PDS), FTIR photoacoustic (FTIR-PAS), FT Raman, and FTIR-attenuated total reflection (FTIR-ATR) spectroscopy as analytical tools for investigating the drug content in semisolid formulations. Regarding the analytical parameters, this study demonstrates the photothermal beam deflection to be definitely comparable to well established spectroscopic methods for this purpose. The correlation coefficients range from 0.990 to 0.999. Likewise, repeatability and limit of detection are comparable.

  5. Identification of functional groups on the surface of modified organic materials using the FT-IR/PAS method

    NASA Astrophysics Data System (ADS)

    Hofman, M.; Pasieczna-Patkowska, S.; Ryczkowski, J.; Wachowski, L.

    2008-02-01

    Interactions between NO and the surface of waste plum stones have been studied by FTIR-PAS in order to identify the nitrogen species formed. It is possible that there are some additional active centres generated on their surface. Identification of surface functional groups was needed to characterize their influence on the properties of obtained materials which were used as catalyst support in our further research.

  6. Television as Textbook: "Cuéntame cómo pasó" in the Spanish (Literature) Classroom

    ERIC Educational Resources Information Center

    Bartlett, Linda; Manyé, Lourdes

    2015-01-01

    The long-running Spanish television program "Cuéntame cómo pasó" represents not only a wildly successful series for Radio Televisión Española, but also an excellent example of the project of historical memory. Premiering in 2001 (but set, in the first season, in 1968), the story of the multigenerational Alcántara family forms a…

  7. Early doors (Edo) mutant mouse reveals the importance of period 2 (PER2) PAS domain structure for circadian pacemaking

    PubMed Central

    Militi, Stefania; Maywood, Elizabeth S.; Sandate, Colby R.; Chesham, Johanna E.; Parsons, Michael J.; Vibert, Jennifer L.; Joynson, Greg M.; Partch, Carrie L.; Hastings, Michael H.; Nolan, Patrick M.

    2016-01-01

    The suprachiasmatic nucleus (SCN) defines 24 h of time via a transcriptional/posttranslational feedback loop in which transactivation of Per (period) and Cry (cryptochrome) genes by BMAL1–CLOCK complexes is suppressed by PER–CRY complexes. The molecular/structural basis of how circadian protein complexes function is poorly understood. We describe a novel N-ethyl-N-nitrosourea (ENU)-induced mutation, early doors (Edo), in the PER-ARNT-SIM (PAS) domain dimerization region of period 2 (PER2) (I324N) that accelerates the circadian clock of Per2Edo/Edo mice by 1.5 h. Structural and biophysical analyses revealed that Edo alters the packing of the highly conserved interdomain linker of the PER2 PAS core such that, although PER2Edo complexes with clock proteins, its vulnerability to degradation mediated by casein kinase 1ε (CSNK1E) is increased. The functional relevance of this mutation is revealed by the ultrashort (<19 h) but robust circadian rhythms in Per2Edo/Edo; Csnk1eTau/Tau mice and the SCN. These periods are unprecedented in mice. Thus, Per2Edo reveals a direct causal link between the molecular structure of the PER2 PAS core and the pace of SCN circadian timekeeping. PMID:26903623

  8. Transient expression of Nxf, a bHLH-PAS transactivator induced by neuronal preconditioning, confers neuroprotection in cultured cells.

    PubMed

    Hester, Ian; McKee, Sarah; Pelletier, Phillip; Thompson, Charles; Storbeck, Christopher; Mears, Alan; Schulz, Jörg B; Hakim, Antoine A; Sabourin, Luc A

    2007-03-01

    Cortical spreading depression (CSD) induces waves of neuronal depolarization that confer neuroprotection to subsequent ischemic events in the rat brain. To gain insights into the molecular mechanisms elicited by CSD, we used representational difference analysis (RDA) to identify mRNAs induced by potassium depolarization in vivo. Using this approach, we have isolated a cDNA encoding the SIM2-related bHLH-PAS protein Nxf. Our results confirm that Nxf mRNA and protein are rapidly and transiently expressed in cortical neurons following CSD. Reporter assays show that Nxf is a transcriptional activator that associates with the bHLH-PAS sub-class co-factor ARNT2. Adenovirus-mediated expression of epitope-tagged Nxf results in cell death and the direct activation of the Bax gene in cultured cells. However, RNA interference studies show that endogenous Nxf is required for optimal neuroprotection by preconditioning in cultured F-11 cells. Together, our data indicate that Nxf is a novel bHLH-PAS transactivator transiently induced by preconditioning and that its sustained expression is detrimental. The identification of Nxf may represent an important step in our understanding of the molecular mechanisms of brain preconditioning and injury. PMID:17214977

  9. The role of ascorbate peroxidase, guaiacol peroxidase, and polysaccharides in cassava (Manihot esculenta Crantz) roots under postharvest physiological deterioration.

    PubMed

    Uarrota, Virgílio Gavicho; Moresco, Rodolfo; Schmidt, Eder Carlos; Bouzon, Zenilda Laurita; Nunes, Eduardo da Costa; Neubert, Enilto de Oliveira; Peruch, Luiz Augusto Martins; Rocha, Miguel; Maraschin, Marcelo

    2016-04-15

    This study aimed to investigate the role of ascorbate peroxidase (APX), guaiacol peroxidase (GPX), polysaccharides, and protein contents associated with the early events of postharvest physiological deterioration (PPD) in cassava roots. Increases in APX and GPX activity, as well as total protein contents occurred from 3 to 5 days of storage and were correlated with the delay of PPD. Cassava samples stained with Periodic Acid-Schiff (PAS) highlighted the presence of starch and cellulose. Degradation of starch granules during PPD was also detected. Slight metachromatic reaction with toluidine blue is indicative of increasing of acidic polysaccharides and may play an important role in PPD delay. Principal component analysis (PCA) classified samples according to their levels of enzymatic activity based on the decision tree model which showed GPX and total protein amounts to be correlated with PPD. The Oriental (ORI) cultivar was more susceptible to PPD. PMID:26617011

  10. Congenital Ichthyosis in 14 Great Dane Puppies With a New Presentation.

    PubMed

    Hoffmann, A; Metzger, J; Wöhlke, A; Peters, M; Junginger, J; Mischke, R; Distl, O; Hewicker-Trautwein, M

    2016-05-01

    The present study describes a generalized congenital skin condition in 14 Great Dane puppies. Macroscopically, all dogs showed generalized gray to yellow scaling and skin wrinkles on the head and all 4 extremities. Skin sections were histologically examined using hematoxylin and eosin, Heidenhain's Azan, and Sudan red III staining methods and by conducting the alcian blue/periodic acid Schiff (AB/PAS) reaction technique on sections. Furthermore, incubation with hyaluronidase was performed. Skin samples were ultrastructurally analyzed using transmission electron microscopy. All affected Great Dane puppies had epidermal and follicular orthokeratotic hyperkeratosis, enlarged keratohyaline granules, vacuolated keratinocytes, and accumulations of an eosinophilic and alcianophilic, lipid-rich material within dilated hair follicular lumina and the cytoplasm of sebocytes. The macroscopic, histopathologic, and ultrastructural skin changes in all 14 Great Dane puppies indicate a new variant of a primary disorder of cornification with congenital, non-epidermolytic, lamellar ichthyosiform appearance. PMID:26242581

  11. Low-dose radiation pretreatment improves survival of human ceiling culture-derived proliferative adipocytes (ccdPAs) under hypoxia via HIF-1 alpha and MMP-2 induction

    SciTech Connect

    Adachi, Naoki; Kubota, Yoshitaka; Kosaka, Kentarou; Akita, Shinsuke; Sasahara, Yoshitarou; Kira, Tomoe; Kuroda, Masayuki; Mitsukawa, Nobuyuki; Bujo, Hideaki; Satoh, Kaneshige

    2015-08-07

    Poor survival is a major problem of adipocyte transplantation. We previously reported that VEGF and MMPs secreted from transplanted adipocytes are essential for angiogenesis and adipogenesis. Pretreatment with low-dose (5 Gy) radiation (LDR) increased VEGF, MMP-2, and HIF-1 alpha mRNA expression in human ceiling culture-derived proliferative adipocytes (hccdPAs). Gene expression after LDR differed between adipose-derived stem cells (hASCs) and hccdPAs. Pretreatment with LDR improved the survival of hccdPAs under hypoxia, which is inevitable in the early stages after transplantation. Upregulation of VEGF and MMP-2 after LDR in hccdPAs is mediated by HIF-1 alpha expression. Our results suggest that pretreatment with LDR may improve adipocyte graft survival in a clinical setting through upregulation of VEGF and MMP-2 via HIF-1 alpha. - Highlights: • Ceiling culture-derived proliferative adipocytes (ccdPAs) react to radiation. • Low-dose radiation (LDR) pretreatment improves survival of ccdPAs under hypoxia. • Gene expression after LDR differs between ccdPAs and adipose-derived stem cells. • LDR-induced increase in MMP-2 and VEGF is dependent on HIF-1 alpha induction. • LDR pretreatment may improve the adipocyte graft survival rate in clinical settings.

  12. A comprehensive protein–protein interactome for yeast PAS kinase 1 reveals direct inhibition of respiration through the phosphorylation of Cbf1

    PubMed Central

    DeMille, Desiree; Bikman, Benjamin T.; Mathis, Andrew D.; Prince, John T.; Mackay, Jordan T.; Sowa, Steven W.; Hall, Tacie D.; Grose, Julianne H.

    2014-01-01

    Per-Arnt-Sim (PAS) kinase is a sensory protein kinase required for glucose homeostasis in yeast, mice, and humans, yet little is known about the molecular mechanisms of its function. Using both yeast two-hybrid and copurification approaches, we identified the protein–protein interactome for yeast PAS kinase 1 (Psk1), revealing 93 novel putative protein binding partners. Several of the Psk1 binding partners expand the role of PAS kinase in glucose homeostasis, including new pathways involved in mitochondrial metabolism. In addition, the interactome suggests novel roles for PAS kinase in cell growth (gene/protein expression, replication/cell division, and protein modification and degradation), vacuole function, and stress tolerance. In vitro kinase studies using a subset of 25 of these binding partners identified Mot3, Zds1, Utr1, and Cbf1 as substrates. Further evidence is provided for the in vivo phosphorylation of Cbf1 at T211/T212 and for the subsequent inhibition of respiration. This respiratory role of PAS kinase is consistent with the reported hypermetabolism of PAS kinase–deficient mice, identifying a possible molecular mechanism and solidifying the evolutionary importance of PAS kinase in the regulation of glucose homeostasis. PMID:24850888

  13. Heparin reduces nonspecific eosinophil staining artifacts in mass cytometry experiments.

    PubMed

    Rahman, Adeeb H; Tordesillas, Leticia; Berin, M Cecilia

    2016-06-01

    The analysis of heterogeneous cell samples by mass cytometry (CyTOF) relies on the assumption that metal labeled antibodies accurately bind to their target antigens. We report a previously unappreciated experimental artifact of non-specific antibody binding by eosinophils during intracellular CyTOF analysis of human whole blood samples. We hypothesized that this non-specific binding results from a charge-based interaction between the metal-labeled antibodies and highly cationic proteins found in eosinophillic granules and found that this non-specific staining artifact could be reduced to background levels with a simple blocking protocol using heparin as a competing anionic protein. This protocol eliminates a potential source of erroneous data interpretation in all experiments involving intracellular staining of human whole blood samples, and allows accurate assessment of dynamic changes in intracellular proteins in eosinophils by CyTOF. © 2016 International Society for Advancement of Cytometry. PMID:27061608

  14. Oxalate films and red stains on Carrara marble.

    PubMed

    Realini, Marco; Colombo, Chiara; Sansonetti, Antonio; Rampazzi, Laura; Colombini, Maria Perla; Bonaduce, Ilaria; Zanardini, Elisabetta; Abbruscato, Pamela

    2005-01-01

    The analytical studies carried out during two different diagnostic surveys, respectively in 1983 and 2003, offered the opportunity to control decay phenomena development on stones facing Certosa of Pavia (Italy). Calcium oxalate films and red stains, present on Carrara marble surface, have been particularly focused; these are the only decay phenomena which apparently have remained unchanged during a period of twenty years. More sensitive and in-depth analytical studies (FTIR equipped with diamond cell, GC-MS, SEM-EDS and optical microscopy) achieved a better knowledge about their composition. Results allowed a critical evaluation of the role of oxalate films on the external marble surface and to suggest new hypotheses about the formation of red stains. PMID:16485663

  15. Lectins stain cells differentially in the coral, Montipora capitata.

    PubMed

    Work, Thierry M; Farah, Yael

    2014-03-01

    A limitation in our understanding of coral disease pathology and cellular pathogenesis is a lack of reagents to characterize coral cells. We evaluated the utility of plant lectins to stain tissues of a dominant coral, Montipora capitata, from Hawaii. Of 22 lectins evaluated, nine of these stained structures in the upper or basal body wall of corals. Specific structures revealed by lectins that were not considered distinct or evident on routine hematoxylin and eosin sections of coral tissues included apical and basal granules in gastrodermis and epidermis, cnidoglandular tract and actinopharynx cell surface membranes, capsules of mature holotrichous isorhizas, and perivitelline and periseminal cells. Plant lectins could prove useful to further our understanding of coral physiology, anatomy, cell biology, and disease pathogenesis. PMID:24518620

  16. Lectins stain cells differentially in the coral, Montipora capitata

    USGS Publications Warehouse

    Work, Thierry M.; Farah, Yael

    2014-01-01

    A limitation in our understanding of coral disease pathology and cellular pathogenesis is a lack of reagents to characterize coral cells. We evaluated the utility of plant lectins to stain tissues of a dominant coral, Montipora capitata, from Hawaii. Of 22 lectins evaluated, nine of these stained structures in the upper or basal body wall of corals. Specific structures revealed by lectins that were not considered distinct or evident on routine hematoxylin and eosin sections of coral tissues included apical and basal granules in gastrodermis and epidermis, cnidoglandular tract and actinopharynx cell surface membranes, capsules of mature holotrichous isorhizas, and perivitelline and periseminal cells. Plant lectins could prove useful to further our understanding of coral physiology, anatomy, cell biology, and disease pathogenesis.

  17. Development of Cell Staining Technique for X-Ray Microscopy

    SciTech Connect

    Tseng, P. Y.; Shih, Y. T.; Liu, C. J.; Hsu, T.; Chien, C. C.; Leng, W. H.; Liang, K. S.; Yin, G. C.; Chen, F. R.; Je, J. H.; Margaritondo, G.; Hwu, Y.

    2007-01-19

    We report a technique for detection of sub-cellular organelles and proteins with hard x-ray microscopy. Several metals were used for enhancing contrast for x-ray microscopy. Osmium tetroxide provides an excellent stain for lipid and can delineate cell membrane. Uranyl acetate has high affinity for nucleotide and can stain nucleus. Immunolocalization of specific proteins and sub-cellular organelles was achieved by 3'3 diaminobenzidine (DAB) with nickel enhancement and nanogold-conjugated secondary antibody with silver enhancement. The x-rays emitted from synchrotron source was monochromatized by double crystal monochromator, the photon energy was fixed at 8 keV to optimize the focusing efficiency of the zone plates. The estimated resolution is about 60 nm. When compared with visible light and conventional confocal microscopy, the X-ray microscopy provides a superior resolution to both conventional optical microscopes.

  18. Cement line staining in undecalcified thin sections of cortical bone

    NASA Technical Reports Server (NTRS)

    Bain, S. D.; Impeduglia, T. M.; Rubin, C. T.

    1990-01-01

    A technique for demonstrating cement lines in thin, undecalcified, transverse sections of cortical bone has been developed. Cortical bone samples are processed and embedded undecalcified in methyl methacrylate plastic. After sectioning at 3-5 microns, cross-sections are transferred to a glass slide and flattened for 10 min. Sections of cortical bone are stained for 20 sec free-floating in a fresh solution of 1% toluidine blue dissolved in 0.1% formic acid. The section is dehydrated in t-butyl alcohol, cleared in xylene, and mounted with Eukitt's medium. Reversal lines appear as thin, scalloped, dark blue lines against a light blue matrix, whereas bone formation arrest lines are thicker with a smooth contour. With this technique cellular detail, osteoid differentiation, and fluorochrome labels are retained. Results demonstrate the applicability of a one-step staining method for cement lines which will facilitate the assessment of bone remodeling activity in thin sections of undecalcified cortical bone.

  19. Angiolymphoid hyperplasia with eosinophilia developing within a port wine stain.

    PubMed

    Manton, Robert N; Itinteang, Tinte; de Jong, Sophie; Brasch, Helen D; Tan, Swee T

    2016-01-01

    A 19-year-old male with a port wine stain on the base of his neck presented with a 5-month history of gradual thickening of the involved skin which interfered with clothing and caused repeated bleeding. The lesion was excised and histopathologic examination revealed angiolymphoid hyperplasia with eosinophilia (ALHE) arising from the pre-existing port wine stain - a rare finding with only one previously reported case. Additionally the lesion was associated with elevated serum renin levels which virtually normalized following excision of the lesion. We further demonstrated the expression of angiotensin converting enzyme and angiotensin II receptors 1 and 2 by the lesion and discuss the possible role of the renin-angiotensin system in this condition. PMID:26010041

  20. Identification of calcium oxalate crystals using alizarin red S stain.

    PubMed

    Proia, A D; Brinn, N T

    1985-02-01

    Calcium oxalate crystals stain with alizarin red S at a pH of 7.0 but not at a pH of 4.2. In contrast, calcium phosphate and calcium carbonate stain at a pH of both 7.0 and 4.2. This difference allows presumptive identification of calcium oxalate deposits. The identity of calcium oxalate can then be confirmed by its insolubility in 2M acetic acid, since both calcium carbonate and calcium phosphate are soluble. We have applied this procedure for several years and have found it to be a rapid, reliable, and technically simple procedure for distinguishing calcium oxalate from other calcium deposits. PMID:2579619

  1. Identifying neutrophils in H&E staining histology tissue images.

    PubMed

    Wang, Jiazhuo; MacKenzie, John D; Ramachandran, Rageshree; Chen, Danny Z

    2014-01-01

    Identifying neutrophils lays a crucial foundation for diagnosing acute inflammation diseases. But, such computerized methods on the commonly used H&E staining histology tissue images are lacking, due to various inherent difficulties of identifying cells in such image modality and the challenge that a considerable portion of neutrophils do not have a "textbook" appearance. In this paper, we propose a new method for identifying neutrophils in H&E staining histology tissue images. We first segment the cells by applying iterative edge labeling, and then identify neutrophils based on the segmentation results by considering the "context" of each candidate cell constructed by a new Voronoi diagram of clusters of other neutrophils. We obtain good performance compared with two baseline algorithms we constructed, on clinical images collected from patients suspected of having inflammatory bowl diseases. PMID:25333103

  2. Identification of active fluorescence stained bacteria by Raman spectroscopy

    NASA Astrophysics Data System (ADS)

    Krause, Mario; Beyer, Beatrice; Pietsch, Christian; Radt, Benno; Harz, Michaela; Rösch, Petra; Popp, Jürgen

    2008-04-01

    Microorganisms can be found everywhere e.g. in food both as useful ingredients or harmful contaminations causing food spoilage. Therefore, a fast and easy to handle analysis method is needed to detect bacteria in different kinds of samples like meat, juice or air to decide if the sample is contaminated by harmful microorganisms. Conventional identification methods in microbiology require always cultivation and therefore are time consuming. In this contribution we present an analysis approach to identify fluorescence stained bacteria on strain level by means of Raman spectroscopy. The stained bacteria are highlighted and can be localized easier against a complex sample environment e.g. in food. The use of Raman spectroscopy in combination with chemometrical methods allows the identification of single bacteria within minutes.

  3. Genetic Variants Associated with Port-Wine Stains

    PubMed Central

    Wooderchak-Donahue, Whitney; Tan, Oon T.; Margraf, Rebecca; Stevenson, David A.; Grimmer, J. Fredrik; Bayrak-Toydemir, Pinar

    2015-01-01

    Background Port-wine stains (PWS) are capillary malformations, typically located in the dermis of the head and neck, affecting 0.3% of the population. Current theories suggest that port-wine stains are caused by somatic mutations that disrupt vascular development. Objectives Understanding PWS genetic determinants could provide insight into new treatments. Methods Our study used a custom next generation sequencing (NGS) panel and digital polymerase chain reaction to investigate genetic variants in 12 individuals with isolated port-wine stains. Importantly, affected and healthy skin tissue from the same individual were compared. A subtractive correction method was developed to eliminate background noise from NGS data. This allowed the detection of a very low level of mosaicism. Results A novel somatic variant GNAQ, c.547C>G, p.Arg183Gly was found in one case with 4% allele frequency. The previously reported GNAQ c.548G>A, p.Arg183Gln was confirmed in 9 of 12 cases with an allele frequency ranging from 1.73 to 7.42%. Digital polymerase chain reaction confirmed novel variants detected by next generation sequencing. Two novel somatic variants were also found in RASA1, although neither was predicted to be deleterious. Conclusions This is the second largest study on isolated, non-syndromic PWS. Our data suggest that GNAQ is the main genetic determinant in this condition. Moreover, isolated port-wine stains are distinct from capillary malformations seen in RASA1 disorders, which will be helpful in clinical evaluation. PMID:26192947

  4. Use of modified Fraser's stain in Promoting Activity Test (PAT).

    PubMed

    Borràs, M

    1988-09-01

    The Promoting Activity Test (PAT) requires a staining procedure that allows rapid, accurate and reliable counting of mitotic figures. We propose use of Fraser's kernechtrot-crystal violet technique, but eliminating the picric-alcoholic differentiation to avoid fading. This modified protocol gives higher mitotic counts in adult mouse adrenal cortex than the hematoxylin-eosin originally used, especially with respect to less conspicuous prophases. PMID:2464217

  5. Method and apparatus for staining immobilized nucleic acids

    DOEpatents

    Ramsey, J. Michael; Foote, Robert S.; Jacobson, Stephen C.

    2000-01-01

    A method for staining immobilized nucleic acids includes the steps of affixing DNA probes to a solid substrate, moving target DNA material into proximity with the DNA probes, whereby the target DNA hybridized with specific ones of the DNA probes, and moving a fluorescent dye into proximity with the hybridized target DNA, whereby the fluorescent dye binds to the hybridized DNA to enable subsequent detection of fluorescence.

  6. Human Salivary Alpha-Amylase (EC.3.2.1.1) Activity and Periodic Acid and Schiff Reactive (PAS) Staining: A Useful Tool to Study Polysaccharides at an Undergraduate Level

    ERIC Educational Resources Information Center

    Fernandes, Ruben; Correia, Rossana; Fonte, Rosalia; Prudencio, Cristina

    2006-01-01

    Health science education is presently in discussion throughout Europe due to the Bologna Declaration. Teaching basic sciences such as biochemistry in a health sciences context, namely in allied heath education, can be a challenging task since the students of preclinical health sciences are not often convinced that basic sciences are clinically…

  7. Chlorination effect on the fluorescence of nucleic acid staining dyes.

    PubMed

    Phe, M H; Dossot, M; Block, J C

    2004-10-01

    An alternative to culture methods for the control of drinking water disinfection would use fluorescent dyes that could evidence the nucleic acid damages provoked by sodium hypochlorite treatment. The two dyes selected in this study, SYBR Green II RNA gel stain and TOTO-1 iodide, efficiently stain nucleic acids (DNA and RNA) and quite poorly the other biomolecules considered (Bovine serum albumin, palmitic acid and dextrane). After treatment of nucleic acid solutions with increasing amounts of sodium hypochlorite, a decrease of fluorescence intensity is observed for both DNA and RNA stained with either SYBR-II or TOTO-1. However, the two fluorochromes do not lead to the same results, which shows that the two dyes are not bound to nucleic acids in the same way. Contrary to TOTO-1, SYBR-II reveals to be sufficiently sensitive to indicate both DNA or RNA damages as soon as the latter are in contact with hypochlorite even at concentrations of HClO lower than 10 micromol/L. Moreover, SYBR-II offers the opportunity to make quantitative titration of chlorine treated DNA and therefore seems to be the appropriate candidate to control the efficiency of the hypochlorite disinfection process of drinking water samples. PMID:15350425

  8. Coffee Stains from Drops with Receding Contact Lines

    NASA Astrophysics Data System (ADS)

    Freed-Brown, Julian

    2015-03-01

    We present a framework for calculating the surface density profile of a coffee stain deposited by a drying drop with a receding contact line. For standard coffee stains, the fluid pins to the substrate, forces flow towards the exterior of the drop and deposits a thin, concentrated ring of particles. Unlike a pinned drop, a receding drop pushes fluid towards its interior and continuously deposits mass across its substrate as it evaporates. This gives rise to a new class of mountain-like morphologies that are not seen in the standard coffee ring effect but are reminiscent of recent experimental results. For a thin, circular drop with uniform evaporation, we calculate the surface density profile analytically and find that it diverges towards the center of the drop as η ~r - 1 / 2 , where r is the distance from the center. We estimate how this divergence is softened due to solute interactions at the final stage of drying. Our framework can easily be extended numerically or analytically to investigate novel stain morphologies left by drying drops of different shapes and evaporation profiles. This work is part of a thesis project advised by Tom Witten. It was supported in part by the National Science Foundation's MRSEC Program under Award Number DMR 0820054.

  9. Pasteurella pestis detection in Fleas by fluorescent antibody staining.

    PubMed

    Hudson, B W; Kartman, L; Prince, F M

    1966-01-01

    In an effort to develop a method for the rapid field identification of plague-infected fleas, the authors have studied the feasibility of direct fluorescent antibody staining of the midgut contents of fleas fed on mice infected with Pasteurella pestis. Fluorescent antibodies prepared from antisera derived from rabbits inoculated with the water-soluble P. pestis fraction 1b antigen, the somatic antigen of heat-killed P. pestis (Bryans strain), and live avirulent (strain A1122) or virulent (Yreka strain) plague vaccines were used used in this study.This direct staining method proved to be impracticable, but encouraging results were obtained by fluorescent antibody staining of broth cultures of macerates of infected fleas after 24-48 hours' incubation.The broth enrichment technique has not yet been evaluated in the field, but it is expected to be of value since it is relatively simple to perform and requires only material that can easily be transported to remote areas. PMID:5328902

  10. Evaluation of immunohistochemical staining for glucagon in human pancreatic tissue

    PubMed Central

    Gurlo, Tatyana; Butle, Peter C.; Butler, Alexandra E.

    2016-01-01

    Immunohistochemistry (IHC) and immunofluorescence (IF) staining techniques are important diagnostic tools of anatomic pathology in the clinical setting and widely used analytical tools in research laboratories. In diabetes research, they are routinely used for the assessment of beta- and alpha-cell mass, for assessment of endocrine cell distribution within the pancreas, for evaluation of islet composition and islet morphology. Here, we present the evaluation of IHC techniques for the detection of alpha-cells in human pancreatic tissue. We compared the Horse Radish Peroxidase (HRP)-based method utilizing DAB Peroxidase Substrate to the Alkaline Phosphatase (AP)-based method utilizing Vector Red substrate. We conclude that HRP–DAB staining is a robust and reliable method for detection of alpha-cells using either rabbit polyclonal or mouse monoclonal anti-glucagon antibodies. However, AP-Vector Red staining should be used with caution, because it is affected by the dehydration with ethanol and toluene preceding the mounting of slides with Permount mounting medium. When AP-Vector Red is a preferable method for alpha-cell labeling, slides should be mounted using aqueous mounting medium or, alternatively, they could be air-dried before permanent mounting PMID:27182095

  11. Accurate PSF-matched photometry for the J-PAS survey

    NASA Astrophysics Data System (ADS)

    Jimenez-Teja, Yolanda; Benitez, Txitxo; Dupke, Renato a.

    2015-08-01

    The Javalambre-PAU Astrophysical Survey (J-PAS) is expected to map 8,600 squared-degrees of the sky using 54 narrow and 5 broad band filters. Carried out by a Spanish-Brazilian consortium, the main goal of this survey is to measure Baryon Acoustic Oscillations (BAOs) with photometric redshifts. The expectation is measuring these photometric redshifts with a precision of dz/(1 + z) ~ 0.003 for 100 million galaxies and a few millions of quasars, and reaching an accuracy of dz/(1 + z) ~ 0.01 for other 300 million galaxies. With these numbers, it will be possible to determine w in the Dark Energy (DE) equation of state with an accuracy of < 4%. To achieve such precision and measure the radial BAOs, not only an advanced technical setup but also special data processing tools are required. These tools must be accurate as well as suitable to be implemented in fully automated and computationally efficient algorithms.The factor that most influences the photometric redshift precision is the quality of the photometry. For that reason we have developed a new technique based on the Chebyshev-Fourier bases (CHEFs, Jiménez-Teja & Benítez 2012, ApJ, 745, 150) to obtain a highly precise multicolor photometry without PSF consideration, thus saving a considerable amount of time and circumventing severe problems such as the PSF variability across the images. The CHEFs are a set of mathematical orthonormal bases with different scale and resolution levels, originally designed to fit the surface light distribution of galaxies. They have proved to be able to model any kind of morphology, including spirals, highly elliptical, or irregular galaxies, including isophotal twists and fine substructure. They also fit high signal-to-noise images, lensing arcs and stars with great accuracy. We can calculate optimal, unbiased, total magnitudes directly through these CHEFs models and, thus, colors without needing the PSF.We compare our photometry with widely-used codes such as SExtractor (Bertin

  12. En bloc staining with hydroquinone treatment for block face imaging.

    PubMed

    Togo, Akinobu; Ohta, Keisuke; Higashi, Ryuhei; Nakamura, Kei-Ichiro

    2014-11-01

    IntroductionBecause recent three-dimensional (3D) ultrastructural reconstruction techniques such as serial block face scanning electron microscopy (SBFSEM), obtain their images directly from the flat surface of specimens via material contrast[1], specimens should be strongly stained with heavy metals prior to resin embedding in order to obtain higher material contrast using backscattered electrons (BSEs). To enhance membrane contrast for block face imaging (BFI), we usually stain specimens using the method published by Deerinck[2], and the images obtained show TEM-like contrast.However, recently, our research subjects have required reconstruction of a much larger volume, increasing the total image acquisition time. To reduce the total acquisition time, both high sensitivity detectors and a new specimen preparation method that provides much higher contrast are required. Takahashi et al.[3] have reported that hydroquinone (HQ) treatment during traditional electro-conductive staining increases specimen conductivity and drastically reduces the charge problem for SEM observation. They concluded that HQ treatment might increase the efficiency of secondary electron (SE) generation. Because BFI can be performed using SE as well as BSE, we examined whether addition of HQ treatment to en bloc staining protocols increased the contrast for BFI using SE. Materials & methodsMouse liver tissue was used. Mice were deeply anesthetized by diethyl ether and sodium pentobarbital, and tissues were fixed by transcardial perfusion of 2% paraformaldehyde and 2.5% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4) through the left ventricle, followed by heparin-containing saline. After perfusion, liver tissues were removed and cut into small cubes approximately 1 mm(3) in the fixative, and were further fixed in the same fixative for 2 h at 4°C. Subsequently, en blocstaining was performed as follows: the specimens were treated using a reduced-OTO staining method (1.5% potassium

  13. Comparative study of the efficacy of Wright-Giemsa stain and Liu's stain in the detection of Auer rods in acute promyelocytic leukemia.

    PubMed

    Yue, Qing Fang; Xiong, Bei; Chen, Wan Xin; Liu, Xin Yue

    2014-07-01

    In view of the importance of Auer rods in the rapid diagnosis of acute promyelocytic leukemia, we compared the results of Wright-Giemsa stain and Liu's stain (a rapid and simple stain, which is also a kind of modified Romanowsky stain) in the detection of Auer rods. This study was based on 53 cases of acute promyelocytic leukemia. Two staining methods were respectively performed on the bone marrow smears of these cases, and presence of Auer rods as well as nuclear features, cytoplasmic features and the degree of granularity of the cytoplasm were compared in each case. Our results showed that the occurrence of Auer rods as well as faggots in leukemic promyelocytes were significantly higher under Liu's stain than under Wright-Giemsa stain. Significant differences also existed in the occurrence of hypergranular cells and cytoplasmic protrusions between smears stained with Liu's stain and Wright-Giemsa stain. Liu's stain is important for the rapid diagnosis of suspicious APL, especially in recognizing Auer rods. PMID:24958342

  14. Immunohistochemical study of the digestive tract of Oligosarcus hepsetus

    PubMed Central

    Vieira-Lopes, Danielle A; Pinheiro, Nadja L; Sales, Armando; Ventura, Adriana; Araújo, Francisco G; Gomes, Iracema D; Nascimento, Aparecida A

    2013-01-01

    AIM: To describe the histology of the digestive tract and to investigate the occurrence of endocrine cells in Oligosarcus hepsetus (O. hepsetus). METHODS: The digestive tract (DT) of O. hepsetus was divided into esophagus, two stomach regions (glandular and non-glandular) and two intestinal regions (anterior and posterior). These specimens were processed by routine histological techniques and stained with hematoxylin-eosin, Gomori’s trichrome, periodic acid Schiff (PAS) and Alcian blue (AB). An immunohistochemical method using avidin-biotin-peroxidase was employed. RESULTS: The esophagus is lined with a non-keratinized stratified squamous epithelium that is reactive to PAS and AB. The stomach has a mucosa lined with a simple columnar epithelium with mucus-secreting cells that are reactive only to PAS. The intestine has a simple columnar epithelium with a brush border and goblet cells that are reactive to PAS and AB. Somatostatin, serotonin and cholecystokinin immunoreactive cells were identified throughout the DT. CONCLUSION: This study revealed adaptations for the species’ diet and showed that the distribution and relative frequency of immunoreactive cells are similar to those of other fish. PMID:23569337

  15. Comparison of Renal Amyloid and Hyaline Glomerulopathy in B6C3F1 Mice: An NTP Retrospective Study.

    PubMed

    Hoane, Jessica S; Johnson, Crystal L; Morrison, James P; Elmore, Susan A

    2016-07-01

    Due to potential misdiagnosis of hyaline glomerulopathy (HG) for amyloidosis, a retrospective study of B6C3F1 mice from the National Toxicology Program (NTP) archives was undertaken to determine whether HG had occurred in prior NTP studies and, if so, whether these 2 glomerular lesions could be routinely discriminated. Kidney slides from 7 amyloid-positive control mice, 2 HG-positive control mice, 3 normal or negative control mice, and 41 potential HG mice (with renal-only deposits previously diagnosed as amyloid) were evaluated using hematoxylin and eosin (H&E), periodic acid Schiff (PAS), Congo red (CR), and Masson's trichrome (MT) stains. Utilizing these techniques, HG was reliably distinguished from amyloidosis. All 41 potential HG mice had glomerular deposits histochemically inconsistent with amyloid; the deposits were PAS positive and CR negative. Four of the 41 mice were selected for transmission electron microscopy of the glomerular deposits; ultrastructurally, the deposits in these animals were consistent with HG and not amyloid. Our findings indicate that HG is a spontaneous lesion in B6C3F1 mice of low occurrence, is commonly misdiagnosed as amyloidosis, and is more likely than amyloid to cause glomerular deposits in mice without evidence of deposits in other tissues. Also, HG can be distinguished from amyloid on H&E evaluation; however, the distinction is improved with use of PAS or CR staining and/or ultraviolet evaluation. PMID:27000376

  16. DAPI staining and fluorescence microscopy techniques for phytoplasmas.

    PubMed

    Andrade, Nancy M; Arismendi, Nolberto L

    2013-01-01

    The 4',6-diamidino-2-phenylindole (DAPI) stain technique is a simple method that was developed for confirming the presence of phytoplasmas in hand-cut or freezing microtome sections of infected tissues. DAPI binds AT-rich DNA preferentially, so that phytoplasmas, localized among phloem cells, can be visualized in a fluorescence microscope. The procedure is quick, easy to use, inexpensive, and can be used as a preliminary or quantitative method to detect or quantify phytoplasma-like bodies in infected plants. PMID:22987410

  17. DNA Stains as Surrogate Nucleobases in Fluorogenic Hybridization Probes.

    PubMed

    Hövelmann, Felix; Seitz, Oliver

    2016-04-19

    The increasing importance assigned to RNA dynamics in cells and tissues calls for probe molecules that enable fluorescence microscopy imaging in live cells. To achieve this goal, fluorescence dyes are conjugated with oligonucleotides so as to provide strong emission upon hybridization with the target molecule. The impressive 10(3)-fold fluorescence intensification observed when DNA stains such as thiazole orange (TO) interact with double-stranded DNA is intriguing and prompted the exploration of oligonucleotide conjugates. However, nonspecific interactions of DNA stains with polynucleotides tend to increase background, which would affect the contrast achievable in live-cell imaging. This Account describes the development of DNA-stain-labeled hybridization probes that provide high signal-to-background. We focus on our contributions in context with related advances from other laboratories. The emphasis will be on the requirements of RNA imaging in live cells. To reduce background, intercalator dyes such as TO were appended to peptide nucleic acid (PNA), which is less avidly recognized by DNA stains than DNA/RNA. Constraining the TO dye as a nucleobase surrogate in "forced intercalation (FIT) probes" improved the target specificity, presumably by helping to prevent unspecific interactions. The enforcement of TO intercalation between predetermined base pairs upon formation of the probe-target duplex provided for high brightness and enabled match/mismatch selectivity beyond stringency of hybridization. We show examples that highlight the use of PNA FIT probes in the imaging of mRNA, miRNA, and lncRNA in living cells. The "FIT approach" was recently extended to DNA probes. Signal brightness can become limiting when low-abundance targets ought to be visualized over cellular autofluorescence. We discuss strategies that further the brightness of signaling by FIT probes. Multilabeling with identical dyes does not solve the brightness issue. To avoid self-quenching, we

  18. [A duplicate staining method for permanent specimen of Trichinella spiralis encapsulated larvae].

    PubMed

    Li, Dan; Yang, Ding; Pi, Ben-Wei; Niu, Li-Na; Zhang, Ying; Wang, Guo-Ying

    2012-04-30

    With single staining method, Trichinella spiralis encapsulated larvae specimens were fixed with formaldehyde alcohol acetic acid fixative solution, and stained with alcohol borax-carmine staining solution (4% borax solution 100 ml, carmine 1 g, and 70% alcohol 100 ml). With duplicate staining, the encapsulated larvae specimens were fixed with formaldehyde alcohol acetic acid fixative solution, and double stained with alcohol borax carmine staining solution and fast green staining solution (fast green 0.1 g, 95% alcohol 100 ml). The results showed that with single staining, it was not clear-cut between the cyst and muscle cells although the larva was differentiable, while with duplicate staining, the larva, cyst and muscle cells were distinguished more clearly. PMID:22908823

  19. Structure-Preserving Color Normalization and Sparse Stain Separation for Histological Images.

    PubMed

    Vahadane, Abhishek; Peng, Tingying; Sethi, Amit; Albarqouni, Shadi; Wang, Lichao; Baust, Maximilian; Steiger, Katja; Schlitter, Anna Melissa; Esposito, Irene; Navab, Nassir

    2016-08-01

    Staining and scanning of tissue samples for microscopic examination is fraught with undesirable color variations arising from differences in raw materials and manufacturing techniques of stain vendors, staining protocols of labs, and color responses of digital scanners. When comparing tissue samples, color normalization and stain separation of the tissue images can be helpful for both pathologists and software. Techniques that are used for natural images fail to utilize structural properties of stained tissue samples and produce undesirable color distortions. The stain concentration cannot be negative. Tissue samples are stained with only a few stains and most tissue regions are characterized by at most one effective stain. We model these physical phenomena that define the tissue structure by first decomposing images in an unsupervised manner into stain density maps that are sparse and non-negative. For a given image, we combine its stain density maps with stain color basis of a pathologist-preferred target image, thus altering only its color while preserving its structure described by the maps. Stain density correlation with ground truth and preference by pathologists were higher for images normalized using our method when compared to other alternatives. We also propose a computationally faster extension of this technique for large whole-slide images that selects an appropriate patch sample instead of using the entire image to compute the stain color basis. PMID:27164577

  20. Modeling the uptake of neutral organic chemicals on XAD passive air samplers under variable temperatures, external wind speeds and ambient air concentrations (PAS-SIM).

    PubMed

    Armitage, James M; Hayward, Stephen J; Wania, Frank

    2013-01-01

    The main objective of this study was to evaluate the performance and demonstrate the utility of a fugacity-based model of XAD passive air samplers (XAD-PAS) designed to simulate the uptake of neutral organic chemicals under variable temperatures, external wind speeds and ambient air concentrations. The model (PAS-SIM) simulates the transport of the chemical across the air-side boundary layer and within the sampler medium, which is segmented into a user-defined number of thin layers. Model performance was evaluated using data for polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) from a field calibration study (i.e., active and XAD-PAS data) conducted in Egbert, Ontario, Canada. With some exceptions, modeled PAS uptake curves are in good agreement with the empirical PAS data. The results are highly encouraging, given the uncertainty in the active air sampler data used as input and other uncertainties related to model parametrization (e.g., sampler-air partition coefficients, the influence of wind speed on sampling rates). The study supports the further development and evaluation of the PAS-SIM model as a diagnostic (e.g., to aid interpretation of calibration studies and monitoring data) and prognostic (e.g., to inform design of future passive air sampling campaigns) tool. PMID:24175752

  1. PAS-1, a protein affinity purified from Ascaris suum worms, maintains the ability to modulate the immune response to a bystander antigen.

    PubMed

    Oshiro, Telma M; Enobe, Cristina S; Araújo, Cláudia A; Macedo, Mahasti S; Macedo-Soares, Maria Fernanda

    2006-04-01

    Helminth infections and parasite components have potent immunomodulatory effects on a host's immune system. In the present study, we investigated the effect of PAS-1, a protein component of Ascaris suum adult worms recognized by a monoclonal antibody (MAIP-1), on humoral and cell-mediated responses to a bystander antigen (ovalbumin [OVA]). MAIP-1 recognized only one of the three polypeptide chains of PAS-1, but neutralized the suppressive effect of the whole worm extract on OVA-specific antibody production. PAS-1 inhibited antibody production against a T-cell-dependent, but not a T-cell-independent, antigen in a dose-dependent way. IgM, IgG1, IgG2b, and also IgE and anaphylactic IgG1 levels were downregulated. In addition, PAS-1 inhibited OVA-specific delayed type hypersensitivity reactions in the footpad of mice, showing a potent immunosuppressive activity on both Th1 and Th2 responses that seems to be mediated by the induction of large amounts of IL-10 and IL-4. Indeed, PAS-1-specific spleen cells secreted sevenfold more IL-10 and threefold more IL-4 than OVA-specific cells in response to in vitro restimulation with the respective antigens. In conclusion, we showed that PAS-1, a single protein component from A. suum, maintains all its immunosuppressive properties. PMID:16519731

  2. Differential response to anodal tDCS and PAS is indicative of impaired focal LTP-like plasticity in schizophrenia.

    PubMed

    Strube, Wolfgang; Bunse, Tilmann; Nitsche, Michael A; Palm, Ulrich; Falkai, Peter; Hasan, Alkomiet

    2016-09-15

    Increasing evidence suggests that neural plasticity impairments, observed in schizophrenia patients, are driven by dysfunctional integration of neural signaling. However, what is less clear is whether this impairment is resultant from a general deficit in plastic induction or whether a specific plastic mechanism is affected. In the current study we aimed to assess whether schizophrenia has a selective impact on focal or non-focal plasticity induction. To pursue this goal we utilized two non-invasive stimulation techniques that differ in the mechanism of long-term potentiation (LTP)-like plasticity induction: focal paired associative stimulation (PAS) and non-focal anodal transcranial direct current stimulation (a-tDCS). 20 schizophrenia patients and 20 matched healthy controls received PAS and a-tDCS in two separate sessions. Cortical excitability and cortical plasticity were assessed by transcranial magnetic stimulation (TMS)-elicited motor evoked potentials (MEP). In both study groups, non-focal a-tDCS resulted in a significant increase of mean MEP magnitude indicating the successful induction of non-focal LTP-like plasticity. In contrast, an increase in mean MEP magnitude following PAS was only observed in the control group, suggesting impaired focal LTP-like plasticity in schizophrenia. Additionally, we observed significantly impaired short-latency intracortical inhibition (SICI) in schizophrenia. This is the first study to comparatively evaluate non-focal and focal plasticity mechanisms in schizophrenia patients. The differential patterns of LTP-like plasticity responses indicate that reduced plasticity in schizophrenia could be ascribed to impairments in spatially and temporally restricted signal integration. This impairment, coupled with an observed reduction of inhibitory circuit efficacy, might further contribute to impairments in coordinating focal signals. PMID:27185738

  3. A gigapixel commercially manufactured cryogenic camera for the J-PAS 2.5m survey telescope

    NASA Astrophysics Data System (ADS)

    Jorden, P. R.; Bastable, M.; Clapp, M.; Darby, S.; Dryer, M.; Eaton, T.; Fenemore-Jones, G.; Jerram, P.; Marin-Franch, A.; Palmer, I.; Pittock, R.; Pool, P.; Rennshaw, R.; Taylor, K.; Waltham, N.; Wheeler, P.

    2012-07-01

    The J-PAS (Javalambre Physics-of-the-Accelerating-Universe Astrophysical Survey) project will perform a five-year survey of the northern sky from a new 2.5m telescope in Teruel, Spain. We describe the design concept of a complete cryogenic camera with a mosaic focal plane and 1.2 gigapixel science array which is to be commercially supplied. The focal plane is contained within a novel liquid-nitrogen-cooled vacuum cryostat, with proximity drive electronics designed to achieve a 4 e- readout noise from the 224-channel CCD system.

  4. Complicated Whipple’s disease and endocarditis following tumor necrosis factor inhibitors

    PubMed Central

    Marth, Thomas

    2014-01-01

    AIM: To test whether treatment with tumor necrosis factor inhibitors (TNFI) is associated with complications of Tropheryma whipplei (T. whipplei) infection. METHODS: Because unexplained arthritis is often the first Whipple’s disease (WD) symptom, patients may undergo treatment with TNFI before diagnosis. This may influence the course of infection with T. whipplei, which causes WD, because host immune defects contribute to the pathogenesis of WD. A literature search and cross referencing identified 19 reports of TNFI treatment prior to WD diagnosis. This case-control study compared clinical data in patients receiving TNFI therapy (group I, n = 41) with patients not receiving TNFI therapy (group II, n = 61). Patients from large reviews served as controls (group III, n = 1059). RESULTS: The rate of endocarditis in patient group I was significantly higher than in patient group II (12.2% in group I vs 1.6% in group II, P < 0.05), and group III (12.2% in group I vs 0.16% in group III, P < 0.01). Other, severe systemic or local WD complications such as pericarditis, fever or specific organ manifestations were increased also in group I as compared to the other patient groups. However, diarrhea and weight loss were somewhat less frequent in patient group I. WD is typically diagnosed with duodenal biopsy and periodic acid Schiff (PAS) staining. PAS-stain as standard diagnostic test had a very high percentage of false negative results (diagnostic failure in 63.6% of cases) in group I. Polymerase chain reaction (PCR) for T. whipplei was more accurate than PAS-stainings (diagnostic accuracy, rate of true positive tests 90.9% for PCR vs 36.4% for PAS, P < 0.01). CONCLUSION: TNFI trigger severe WD complications, particularly endocarditis, and lead to false-negative PAS-tests. In case of TNFI treatment failure, infection with T. whipplei should be considered. PMID:25548618

  5. Evaluation of a fluorescent lectin-based staining technique for some acidophilic mining bacteria.

    PubMed

    Fife, D J; Bruhn, D F; Miller, K S; Stoner, D L

    2000-05-01

    A fluorescence-labeled wheat germ agglutinin staining technique (R. K. Sizemore et al., Appl. Environ. Microbiol. 56:2245-2247, 1990) was modified and found to be effective for staining gram-positive, acidophilic mining bacteria. Bacteria identified by others as being gram positive through 16S rRNA sequence analyses, yet clustering near the divergence of that group, stained weakly. Gram-negative bacteria did not stain. Background staining of environmental samples was negligible, and pyrite and soil particles in the samples did not interfere with the staining procedure. PMID:10788401

  6. Antibody Staining in C. Elegans Using "Freeze-Cracking"

    PubMed Central

    Duerr, Janet S.

    2013-01-01

    To stain C. elegans with antibodies, the relatively impermeable cuticle must be bypassed by chemical or mechanical methods. "Freeze-cracking" is one method used to physically pull the cuticle from nematodes by compressing nematodes between two adherent slides, freezing them, and pulling the slides apart. Freeze-cracking provides a simple and rapid way to gain access to the tissues without chemical treatment and can be used with a variety of fixatives. However, it leads to the loss of many of the specimens and the required compression mechanically distorts the sample. Practice is required to maximize recovery of samples with good morphology. Freeze-cracking can be optimized for specific fixation conditions, recovery of samples, or low non-specific staining, but not for all parameters at once. For antibodies that require very hard fixation conditions and tolerate the chemical treatments needed to chemically permeabilize the cuticle, treatment of intact nematodes in solution may be preferred. If the antibody requires a lighter fix or if the optimum fixation conditions are unknown, freeze-cracking provides a very useful way to rapidly assay the antibody and can yield specific subcellular and cellular localization information for the antigen of interest. PMID:24145964

  7. Color stability of ceramic brackets immersed in potentially staining solutions

    PubMed Central

    Guignone, Bruna Coser; Silva, Ludimila Karsbergen; Soares, Rodrigo Villamarim; Akaki, Emilio; Goiato, Marcelo Coelho; Pithon, Matheus Melo; Oliveira, Dauro Douglas

    2015-01-01

    OBJECTIVE: To assess the color stability of five types of ceramic brackets after immersion in potentially staining solutions. METHODS: Ninety brackets were divided into 5 groups (n = 18) according to brackets commercial brands and the solutions in which they were immersed (coffee, red wine, coke and artificial saliva). The brackets assessed were Transcend (3M/Unitek, Monrovia, CA, USA), Radiance (American Orthodontics, Sheboygan, WI, USA), Mystique (GAC International Inc., Bohemia, NY, USA) and Luxi II (Rocky Mountain Orthodontics, Denver, CO, USA). Chromatic changes were analyzed with the aid of a reflectance spectrophotometer and by visual inspection at five specific time intervals. Assessment periods were as received from the manufacturer (T0), 24 hours (T1), 72 hours (T2), as well as 7 days (T3) and 14 days (T4) of immersion in the aforementioned solutions. Results were submitted to statistical analysis with ANOVA and Bonferroni correction, as well as to a multivariate profile analysis for independent and paired samples with significance level set at 5%. RESULTS: The duration of the immersion period influenced color alteration of all tested brackets, even though these changes could not always be visually observed. Different behaviors were observed for each immersion solution; however, brackets immersed in one solution progressed similarly despite minor variations. CONCLUSIONS: Staining became more intense over time and all brackets underwent color alterations when immersed in the aforementioned solutions. PMID:26352842

  8. Analysis of surface stains on modern gold coins

    NASA Astrophysics Data System (ADS)

    Corregidor, V.; Alves, L. C.; Cruz, J.

    2013-07-01

    It is a mandatory practice in the European Mint Houses to provide a certificate of guarantee of their products specially when issuing commemorative gold or silver coins. This practise should assure satisfaction and trust both for the mint house and for the demanding numismatic collector. For these reasons the Mint Houses follow a strict quality control in all the production steps in order to ensure a no-defect, fully supervised output. In spite of all the undertaken precautions, different surface stains with diverse origin on gold coins recently minted in Europe were observed. Those were compositionally studied by means of IBA techniques at the end-stage nuclear microprobe installed at IST/ITN. From this study it was possible to identify several possible sources for these stains. The presence of defects at the surface of these commemorative coins address the need of improving the quality control system and the results here presented point out where these improvements should occur, in order to reduce/eliminate them and give the customer a product that with time probably will be revalued.

  9. Coproduction of detergent compatible bacterial enzymes and stain removal evaluation.

    PubMed

    Niyonzima, Francois N; More, Sunil S

    2015-10-01

    Most of the detergents that are presently produced contain the detergent compatible enzymes to improve and accelerate the washing performance by removing tough stains. The process is environment friendly as the use of enzymes in the detergent formulation reduces the utilization of toxic detergent constituents. The current trend is to use the detergent compatible enzymes that are active at low and ambient temperature in order to save energy and maintain fabric quality. As the detergent compatible bacterial enzymes are used together in the detergent formulation, it is important to co-produce the detergent enzymes in a single fermentation medium as the enzyme stability is assured, and production cost gets reduced enormously. The review reports on the production, purification, characterization and application of detergent compatible amylases, lipases, and proteases are available. However, there is no specific review or minireview on the concomitant production of detergent compatible amylases, lipases, and proteases. In this minireview, the coproduction of detergent compatible enzymes by bacterial species, enzyme stability towards detergents and detergent components, and stain release analysis were discussed. PMID:26011283

  10. A Comparison of Acquired Port-wine Stain with Congenital Port-wine Stain Using an Image Analyzer

    PubMed Central

    Lee, Jung Ju; Lee, Jae Chul; Kim, Byung Soo; Lee, Weon Ju; Kim, Do Won; Jang, Yun Hwan; Bae, Han Ik

    2008-01-01

    Background Recent reports have proposed that there were no differences between acquired port-wine stain (APWS) and congenital port-wine stain (CPWS) except the onset of disease. Pulsed dye laser (PDL) therapy is regarded as the treatment of choice in PWS. Although in some articles, APWS might have shown a better response to PDL than CPWS, this is still controversial. It has been assumed however, that there might be some differences determining therapeutic responses between the two entities. Objective The purpose of this study is to find out some histopathologic differences between APWS and CPWS. Methods 14 patients with APWS and 17 patients with CPWS from our patient files were included in this study. Immunohistochemical staining by factor VIII-related antigen was carried out on the specimens of punch biopsy to better visualize the blood vessels. Histopathologic assessment of variables such as vessel area, percentage of vascular area and vessel depth was performed using a computer-assisted image analyzer program. Results The mean vessel area in APWS was 1014.7 ± 782.5µm2 and that of CPWS was 1341.5 ± 689.9µm2. The mean percentage of vascular area in APWS was 2.02 ± 1.38% and that of CPWS was 2.65 ± 1.56%. The mean vessel depth in APWS was 327.5 ± 120.7µm and 321.7 ± 93.1µm in CPWS. No histopathologic variable was statistically significant using the Mann-Whitney test (p>0.05). PMID:27303148

  11. Reliability of the ICD 10 version of the Psychiatric Assessment Schedule for Adults with Developmental Disability (PAS-ADD).

    PubMed

    Costello, H; Moss, S; Prosser, H; Hatton, C

    1997-08-01

    The Psychiatric Assessment Schedule for Adults with Developmental Disability (PAS-ADD) is a semi-structured clinical interview designed for use with respondents who have learning disability. The first version was based on the Present State Examination. The revised version was derived from the Schedules for Clinical Assessment in Neuropsychiatry (SCAN), and makes ICD 10 diagnoses using the SCAN diagnostic program. This current version has a 4-point scale of severity, compared with the 3-point scale of the first version. It also has a new module relating to psychotic disorders. The sample consisted of 40 individuals representing a spectrum of neurotic, depressive and psychotic disorders. Videotapes of 40 PAS-ADD interviews were re-rated by trained interviewers who had not been involved in the original study in which the videotapes were produced. The mean Kappa across all individual item codes was 0.65, ranging from 0.94 to 0.35. The mean Kappa agreement on item groups was 0.66. Correlation between total symptom scores was 0.74. Agreement on index of definition was Kappa 0.70. We concluded that, agreement was generally lower than for the ICD 9 version. This was probably due mainly to the increase in the severity categories from three to four. However, the new items (most of which related to psychosis) were of comparable reliability to other items. PMID:9299928

  12. Characterization of npas3, a novel basic helix-loop-helix PAS gene expressed in the developing mouse nervous system.

    PubMed

    Brunskill, E W; Witte, D P; Shreiner, A B; Potter, S S

    1999-11-01

    Here we describe the cloning and expression pattern of a new bHLH-PAS domain gene, Npas3. Npas3 shares 50.2% amino acid sequence identity with Npas1 and a lesser similarity with other members of the bHLH-PAS domain family of transcription factors. Northern blot analysis detected Npas3 mRNA between 11.5 and 17.5 d.p.c. in embryonic development and exclusively in the adult brain. Whole-mount and section in situ hybridization assays revealed expression of Npas3 between 9.5 and 11.5 d.p.c. in the developing neural tube. In addition, Npas3 mRNA was expressed throughout the neuroepithelium of the developing central nervous system between 10. 5 and 12.5 d.p.c. Interestingly, at 14.5 d.p.c., the expression of Npas3 mRNA became restricted to the neopallial layer of the cortex. At 12.5 d.p.c., Npas3 mRNA was evident in nonneural tissues such as the developing dermis and mesenchyme surrounding the otic and nasal placodes. Expression was also detected in the developing cardiac valves, limb and developing kidney. PMID:10534623

  13. No Evidence for Interference of Hematoxylin and Eosin (HE) Staining in DNA Testing: Utility of DNA Extraction from HE-Stained Archival Tissue Sections

    PubMed Central

    Morikawa, Teppei; Shima, Kaori; Kuchiba, Aya; Yamauchi, Mai; Tanaka, Noriko; Imamura, Yu; Liao, Xiaoyun; Qian, Zhi Rong; Brahmandam, Mohan; Longtine, Janina A.; Lindeman, Neal I.; Fuchs, Charles S.; Ogino, Shuji

    2012-01-01

    Although histochemical staining has been believed to inhibit DNA amplification reaction, no previous study has systematically evaluated the influence of histochemical staining on downstream molecular assays. To evaluate an influence of hematoxylin and eosin (HE) staining on DNA testing, we isolated DNA from 10 unstained, 10 hematoxylin-stained, 10 eosin-stained or 10 HE-stained tissue sections (ie, 4 groups), from each of 5 colon cancers. Among those 4 groups, we did not observe any significant or appreciable difference in DNA fragmentation by agarose gel electrophoresis; in DNA amplification by real-time PCR; in microsatellite PCR fragment analyses; or in PCR-Pyrosequencing. As a proof-of-principle study, we successfully performed microsatellite instability analysis and sequencing of KRAS and BRAF on over 1300 colorectal cancers using DNA extracted from HE stained tissue sections. Our data provide no evidence for interfering effect of HE staining on DNA testing, suggesting that DNA from HE-stained sections can be effectively used for routine DNA testing. PMID:22706867

  14. New Role of Nod Proteins in Regulation of Intestinal Goblet Cell Response in the Context of Innate Host Defense in an Enteric Parasite Infection

    PubMed Central

    Wang, Huaqing; Kim, Janice J.; Denou, Emmanuel; Gallagher, Amanda; Thornton, David J.; Shajib, M. Sharif; Xia, Lijun; Schertzer, Jonathan D.; Grencis, Richard K.; Philpott, Dana J.

    2015-01-01

    Mucins secreted by intestinal goblet cells are considered an important component of innate defense in a number of enteric infections, including many parasitic infections, but also likely provide protection against the gut microbiota. Nod proteins are intracellular receptors that play key roles in innate immune response and inflammation. Here, we investigated the role of Nod proteins in regulation of intestinal goblet cell response in naive mice and mice infected with the enteric parasite Trichuris muris. We observed significantly fewer periodic acid-Schiff (PAS)-stained intestinal goblet cells and less mucin (Muc2) in Nod1 and Nod2 double-knockout (Nod DKO) mice after T. muris infection than in wild-type (WT) mice. Expulsion of parasites from the intestine was significantly delayed in Nod DKO mice. Treatment of naive WT mice with Nod1 and Nod2 agonists simultaneously increased numbers of PAS-stained goblet cells and Muc2-expressing cells, whereas treatment with Nod1 or Nod2 separately had no significant effect. Stimulation of mucin-secreting LS174T cells with Nod1 and Nod2 agonists upregulated core 3 β1,3-N-acetylglucosaminyltransferase (C3GnT; an important enzyme in mucin synthesis) and MUC2. We also observed lower numbers of PAS-stained goblet cells and less Muc2 in germfree mice. Treatment with Nod1 and Nod2 agonists enhanced the production of PAS-stained goblet cells and Muc2 in germfree mice. These data provide novel information on the role of Nod proteins in goblet cell response and Muc2 production in relation to intestinal innate defense. PMID:26527214

  15. Cutaneous macroglobulinosis: a case series.

    PubMed

    Camp, Brendan J; Magro, Cynthia M

    2012-10-01

    Cutaneous macroglobulinosis is a rare skin manifestation of Waldenstrom macroglobulinemia. It is characterized by the deposition of eosinophilic, immunoglobulin-derived material in the dermis. It typically presents as pink or skin-colored papules favoring the extensor surfaces of the extremities. There are 11 reported cases of cutaneous macroglobulinosis in the literature. In our consultative dermatopathology practice we encountered three additional cases. In Case 1, a 41-year-old female with a monoclonal immunoglobulin M (IgM)-kappa gammopathy developed skin-colored papules on her extensor extremities, forehead and back. Histopathologic and immunohistochemical analyses revealed periodic acid-Schiff (PAS)-positive pink material in the dermis that stained with IgM. In Case 2, an 83-year-old female with a monoclonal IgM-lambda paraproteinemia developed non-blanching papules and plaques on the lower extremities and cheeks. Skin biopsies showed a striking occlusion of the vessels with a PAS-positive eosinophilic precipitate, which was also found in the extravascular spaces and stained with IgM using direct immunofluorescence (DIF) and immunohistochemical stains. In Case 3, an 80-year-old male with Waldenstrom macroglobulinemia developed ulcerated papules and nodules on the lower extremities. The biopsy findings were similar to those of Case 2. We present a series of three patients with cutaneous macroglobulinosis and explore variations in the clinical and histopathological findings of this uncommon entity. PMID:22882527

  16. Morpho-histological characterisation of the alimentary canal of an important food fish, Asian seabass (Lates calcarifer)

    PubMed Central

    Purushothaman, Kathiresan; Lau, Doreen; Saju, Jolly M.; Musthaq SK, Syed; Lunny, Declan Patrick

    2016-01-01

    Asian seabass (Lates calcarifer) is a food fish of increasing aquaculture importance. In order to improve our understanding on the digestive system and feeding of this species, morphological and histological features of the gut were studied. Morphologically, the Asian seabass gut is defined by a short and muscular esophagus, well-developed stomach and comparatively short intestine. Mucous secreting goblet cells reactive to PAS (Periodic Acid Schiff) and AB (Alcian Blue) stain were present throughout the esophagus. The stomach was sac-like and could be distinguished into the cardiac, fundic and pyloric regions. Gastric glands and mucus cells were predominately present in the cardiac and fundic regions. Five finger-like pyloric caeca were present between the stomach and intestine. The intestine was a short, tubular structure with no morphological differences between the various regions. Histologically, the intestinal regions were similar, the main difference being in the number of goblet cells that increased from anterior to posterior intestine, with 114 ± 9, 153 ± 7 and 317 ± 21 goblet cells in the anterior, mid and posterior regions, respectively. The intestinal epithelium stained positively for PAS, but the staining was stronger for acidic glycoproteins. The rectum was similar to intestine, except for increased goblet cell numbers (anterior rectum: 529 ± 26; posterior rectum: 745 ± 29). Gut morpho-histology did not respond to salinity changes, however, there was a significant reduction of mucosal height, goblet cell numbers and muscularis thickness upon food deprivation.

  17. Histochemical characterisation of complex carbohydrates expressed in the alimentary tract of chickens.

    PubMed

    Scocco, Paola; Pedini, Vera

    2010-08-01

    This study characterised the composition of surface and secretion complex carbohydrates in the alimentary tract of organically and intensively housed chickens. Histochemical labelling was carried out on samples of oesophagus, glandular stomach and proximal duodenum using (1) alcian blue (AB) at pH 2.5 combined with neuraminidase digestion (Sial) with or without saponification (KOH), (2) AB at pH 1 and 0.5, (3) periodic acid-Schiff (PAS), (4) a combined AB-PAS stain, and (5) both low and high iron diamine stains combined with enzymatic treatments specific for glycosaminoglycans. Semi-quantitative scoring of the histochemical staining indicated abundant acidic sulfated glycoconjugates in each alimentary tract region, in addition to the presence of sialoglycoderivatives in the oesophagus. These complex carbohydrate components are likely to have various functions including the provision of a protective visco-elastic barrier and a role in the defence against specific pathogens. The findings provide a starting point for a more in-depth study of alimentary tract glycoconjugates in poultry. PMID:19477664

  18. Intestinal histoplasmosis in immunocompetent adults

    PubMed Central

    Zhu, Lin-Lin; Wang, Jin; Wang, Zi-Jing; Wang, Yi-Ping; Yang, Jin-Lin

    2016-01-01

    AIM: To present a retrospective analysis of clinical and endoscopic features of 4 cases of immunocompetent hosts with intestinal histoplasmosis (IH). METHODS: Four immunocompetent adults were diagnosed with IH between October 2005 and March 2015 at West China Hospital of Sichuan University. Clinical and endoscopic characteristics were summarized and analyzed retrospectively. GMS (Gomori methenamine silver), PAS (periodic acid-Schiff) and Giemsa staining technique were used to confirm Histoplasma capsulatum(H. capsulatum). The symptoms, signs, endoscopic presentations, radiographic imaging, pathological stain results and follow-up are presented as tables and illustrations. RESULTS: The cases were male patients, ranging from 33 to 61 years old, and primarily presented with non-specific symptoms such as irregular fever, weight loss, abdominal pain and distention. Hepatosplenomegaly and lymphadenopathy were the most common signs. Endoscopic manifestations were localized or diffuse congestion, edema, ulcers, and polypoid nodules with central erosion involving the terminal ileum, ascending colon, transverse colon, descending colon, sigmoid colon and rectum, similar to intestinal tuberculosis, tumor, and inflammatory bowel disease. Numerous yeast-like pathogens testing positive for PAS and GMS stains but negative for Giemsa were detected in the cytoplasm of the histiocytes, which were highly suggestive of H. capsulatum. CONCLUSION: Immunocompetent individuals suffering from histoplasmosis are rarely reported. It is necessary that gastroenterologists and endoscopists consider histoplasmosis as a differential diagnosis, even in immunocompetent patients. PMID:27099446

  19. Effects of matrix metalloproteinase inhibitor on LPS-induced goblet cell metaplasia.

    PubMed

    Kim, Je Hyeong; Lee, Sung Yong; Bak, Sang Myeon; Suh, In Bum; Lee, Sang Yeub; Shin, Chol; Shim, Jae Jeong; In, Kwang Ho; Kang, Kyung Ho; Yoo, Se Hwa

    2004-07-01

    Bacterial infections of the lung are known to induce inflammatory responses, which lead to mucus hypersecretion. Moreover, mucin synthesis in the airways has been reported to be regulated by neutrophilic inflammation-induced epidermal growth factor receptor (EGFR) expression and its activation. Furthermore, matrix metalloproteinases (MMPs), especially MMP-9, have been reported to promote the transmigration of activated neutrophils. In this study, we investigated the associations between lipopolysaccharide (LPS)-induced goblet cell (GC) metaplasia and EGFR expression and the effects of MMP inhibitor (MMPI). Various concentrations of LPS were instilled into the tracheas of pathogen-free Sprague-Dawley rats, and airways were examined at different times after LPS instillation. To examine the role of MMP-9, we treated rats 3 days before LPS instillation and daily thereafter with MMPI. Neutrophilic infiltration, Alcian blue/periodic acid-Schiff (AB/PAS) staining, and immunohistochemical staining for MUC5AC, EGFR, and MMP-9 were performed. The instillation of LPS increased AB/PAS and MUC5AC staining in time- and dose-dependent manners, and treatment with MMPI significantly prevented GC metaplasia. The instillation of LPS into the trachea also induced neutrophilic infiltration and EGFR and MMP-9 expression in the airway epithelium, and MMPI was found to significantly prevent neutrophil recruitment, GC metaplasia, and EGFR and MMP-9 expression. This study demonstrates that the MMP-9 and EGFR cascades are associated with LPS-induced mucus hypersecretion. PMID:15020297

  20. Gastroprotective Activity of Polygonum chinense Aqueous Leaf Extract on Ethanol-Induced Hemorrhagic Mucosal Lesions in Rats

    PubMed Central

    Ismail, Iza Farhana; Abdul Majid, Nazia; Kadir, Farkaad A.; Al-Bayaty, Fouad; Awang, Khalijah

    2012-01-01

    Polygonum chinense is a Malaysian ethnic plant with various healing effects. This study was to determine preventive effect of aqueous leaf extract of P. chinense against ethanol-induced gastric mucosal injury in rats. Sprague Dawley rats were divided into seven groups. The normal and ulcer control groups were orally administered with distilled water. The reference group was orally administered with 20 mg/kg omeprazole. The experimental groups received the extracts 62.5, 125, 250, and 500 mg/kg, accordingly. After sixty minutes, distilled water and absolute ethanol were given (5 mL/kg) to the normal control and the others, respectively. In addition to histology, immunohistochemical and periodic acid schiff (PAS) stains, levels of lipid peroxidation, malondialdehyde (MDA), antioxidant enzymes, and superoxide dismutase (SOD) were measured. The ulcer group exhibited severe mucosal damages. The experimental groups significantly reduced gastric lesions and MDA levels and increased SOD level. Immunohistochemistry of the experimental groups showed upregulation and downregulation of Hsp70 and Bax proteins, respectively. PAS staining in these groups exhibited intense staining as compared to the ulcer group. Acute toxicity study revealed the nontoxic nature of the extract. Our data provide first evidence that P. chinense extract could significantly prevent gastric ulcer. PMID:23365597

  1. Sizing of single fluorescently stained DNA fragments by scanning microscopy

    PubMed Central

    Laib, Stephan; Rankl, Michael; Ruckstuhl, Thomas; Seeger, Stefan

    2003-01-01

    We describe an approach to determine DNA fragment sizes based on the fluorescence detection of single adsorbed fragments on specifically coated glass cover slips. The brightness of single fragments stained with the DNA bisintercalation dye TOTO-1 is determined by scanning the surface with a confocal microscope. The brightness of adsorbed fragments is found to be proportional to the fragment length. The method needs only minute amount of DNA, beyond inexpensive and easily available surface coatings, like poly-l-lysine, 3-aminoproyltriethoxysilane and polyornithine, are utilizable. We performed DNA-sizing of fragment lengths between 2 and 14 kb. Further, we resolved the size distribution before and after an enzymatic restriction digest. At this a separation of buffers or enzymes was unnecessary. DNA sizes were determined within an uncertainty of 7–14%. The proposed method is straightforward and can be applied to standardized microtiter plates. PMID:14602931

  2. Pulsed photothermal radiometry of port-wine-stain lesions.

    PubMed

    Jacques, S L; Nelson, J S; Wright, W H; Milner, T E

    1993-05-01

    Pulsed photothermal radiometry is used to map the heat deposition in human skin after a short laser pulse. It uses an IR (HgCdTe) detector for a rapid noncontact measurement of the skin surface temperature based on the blackbody emission in the 8-12-microm spectrum. The heat deposited by the laser pulse in the superficial epidermis causes an immediate temperature jump, and the heat deposited in basal epidermal melanin and deep port wine stains diffuses to the surface before detection. The time course of the surface temperature T(z = 0, t), indicates the initial spatial distribution of heat, T(z, t = 0), deposited by the laser. PMID:20820403

  3. X-gal Staining on Adult Mouse Brain Sections

    PubMed Central

    Kokubu, Hiroshi; Lim, Janghoo

    2016-01-01

    Knowing expression patterns of given proteins is very important to understand their functions. Immunostaining analysis with specific antibodies is commonly used to identify cells or tissues expressing proteins of interest. Although this technique is regularly used, it requires high quality of specific antibodies and there is no good quality of antibody available for certain proteins. Alternatively, X-gal staining is also used to analyze protein expression pattern. It is simple and routinely used to detect expression pattern of any proteins of interest in vivo. In this method, genetically modified animals that express beta-galactosidase under the control of certain regulatory elements will be used to reveal the expression pattern of proteins that use the same regulatory elements.

  4. Effect of droplet shape on ring stains from dried liquid

    NASA Astrophysics Data System (ADS)

    Santiago, Melvin; Brown, Katherine; Mathur, Harsh

    A landmark experimental paper on coffee stains by Deegan et al included a simple theoretical analysis of circular droplets. The analysis was based on a model informally called the Maxwell House equations. It describes the evolving height profile of the droplet, the evaporation of the solvent and the outflow of solute to the rim of the droplet. Since typical droplets are not circles, here we extend the analysis to more general shapes. We find that for thin droplets the height profile may be determined by solving Poisson's equation in a domain corresponding to the footprint of the droplet. Evaporation is treated in a simple approximation via an electrostatic analogy and is dominated by the sharp edges of the droplet. Assuming zero vorticity allows us to analyze the solvent flow in droplets of arbitrary shape. We compare circular droplets to other shapes including long linear droplets, ring shaped droplets and droplets with an elliptical footprint

  5. Fat tissue staining and photodynamic/photothermal effects

    NASA Astrophysics Data System (ADS)

    Tuchin, Valery V.; Altshuler, Gregory B.; Yanina, Irina Yu.; Kochubey, Vyacheslav I.; Simonenko, Georgy V.

    2010-02-01

    Cellulite is considered as a disease of the subcutaneous fat layer that appears mostly in women and consists of changes in fat cell accumulation together with disturbed lymphatic drainage, affecting the external appearance of the skin. The photodynamic and selective photothermal treatments may provide reduction the volume of regional or sitespecific accumulations of subcutaneous adipose tissue on the cellular level. We hypothesize that light irradiation of stained fat tissue at selected temperature leads to fat cell lypolytic activity (the enhancement of lipolysis of cell triglycerides due to expression of lipase activity and cell release of free fat acids (FFAs) due to temporal cell membrane porosity), and cell killing due to apoptosis caused by the induced fat cell stress and/or limited cell necrosis.

  6. Identification of dentin phosphophoryn localization by histochemical stainings.

    PubMed

    Takagi, Y; Fujisawa, R; Sasaki, S

    1986-01-01

    Phosphophoryn, the most abundant of the dentin non-collagenous proteins, has been considered to be related in function to the mineralization process. In the present study, identification of dentin phosphophoryn localization was attempted using newly developed, precautionary histological methods by which phosphophoryn was retained in the sections during the specimen preparation and stained selectively in situ. Phosphophoryn was found to be present widely in all of the calcified dentin except the mantle dentin, the external, first-formed portion of dentin, but was not found in the predentin, the inner, uncalcified layer of dentin. These results indicate that phosphophoryn is apparently related to the mineral phase of calcified dentin and that the mineralization process of mantle dentin, which is formed before the odontoblasts are fully differentiated, may be different from that of circumpulpal dentin. PMID:2421974

  7. Preliminary oxidation in histochemical staining methods for cholesterol.

    PubMed

    Adams, C W; High, O B

    1980-08-01

    The need for preliminary oxidation with histochemical methods for cholesterol was investigated on silica-coated sheets and in tissue sections. The techniques used were the Schultz reaction, perchloric acid-naphthoquinone (PAN), Lewis & Lobban's ferric alum-sulphuric acid reagent and Okamoto's iodine-sulphuric acid. The oxidants assessed were ferric chloride, ferric alum, potassium permanganate, ammonium sulphamate and ultraviolet light. The best combinations amongst those tested in order of reactivity were FeCl3-PAN, ferric alum-Schultz, Lewis-Lobban (no additional oxidant), iodine-sulphuric acid (no additional oxidant). Authentic preparations of cholesterol oxidation products were stained with these methods, but the nature of the oxidized product in the preliminary stage could not be determined. PMID:6157826

  8. Ultrastructure, ZIO-staining and chromaffinity of gerbil pinealocytes.

    PubMed

    Chau, Y P; Liao, K K; Kao, M H; Huang, B N; Kao, Y S; Lu, K S

    1994-11-01

    The ultrastructure and cytochemistry of the gerbil pineal gland were studied by the conventional electron microscopy, zinc iodide-osmium tetroxide (ZIO) staining and chromaffin reaction. Conventional electron microscopy revealed that the ultrastructure of gerbil pinealocytes are similar to other rodents, i.e., irregular cell contour with numerous cytoplasmic processes, round or oval nucleus and prominent nucleoli, elongated mitochondria with flattened and tubular cristae and dense matrix, well-developed Golgi apparatus and its associated structures, abundant elements of endoplasmic reticulum--both smooth and rough varieties, and bundles of microfilament and microtubule in the cytoplasm. Some pinealocyte processes contain numerous small clear and "slightly coated" vesicles. Numerous profiles of varicosities containing small dense-cored and clear vesicles were frequently encountered. After ZIO treatment, ZIO staining was preferentially localized in the cytoplasm of some, but not all, of the gerbil pinealocytes. Numerous small clear vesicles (30-50 nm in diameter) in the process of the pinealocytes or in the varicosities of the nerve fibers showed strong ZIO-philia. After chromaffin reaction treatment, the number and electron density of small clear and dense-cored vesicles in the profiles of nerve varicosities increased and this indicates that some of the small clear and dense-cored vesicles in the varicosities are reactive. It is thus concluded that (1) the vesicles in the pinealocytes may be rich in cystine and/or cysteine and possibly the organelle is involved in the sequestering calcium ion during the calcification of the pineal concretions, and (2) the small dense-cored and clear vesicles in the nerve fibers in the gerbil pineal parenchyma may contain both serotonin and primary biogenic amines. PMID:7530780

  9. Multi-stained whole slide image alignment in digital pathology

    NASA Astrophysics Data System (ADS)

    Déniz, Oscar; Toomey, David; Conway, Catherine; Bueno, Gloria

    2015-03-01

    In Digital Pathology, one of the most simple and yet most useful feature is the ability to view serial sections of tissue simultaneously on a computer monitor. This enables the pathologist to evaluate the histology and expression of multiple markers for a patient in a single review. However, the rate limiting step in this process is the time taken for the pathologist to open each individual image, align the sections within the viewer, with a maximum of four slides at a time, and then manually move around the section. In addition, due to tissue processing and pre-analytical steps, sections with different stains have non-linear variations between the two acquisitions, that is, they will stretch and change shape from section to section. To date, no solution has come close to a workable solution to automatically align the serial sections into one composite image. This research work address this problem to obtain an automated serial section alignment tool enabling the pathologists to simply scroll through the various sections in a single viewer. To this aim a multi-resolution intensity-based registration method using mutual information as a similarity metric, an optimizer based on an evolutionary process and a bilinear transformation has been used. To characterize the performance of the algorithm 40 cases x 5 different serial sections stained with hematoxiline-eosine (HE), estrogen receptor (ER), progesterone receptor (PR), Ki67 and human epidermal growth factor receptor 2 (Her2), have been considered. The qualitative results obtained are promising, with average computation time of 26.4s for up to 14660x5799 images running interpreted code.

  10. Quantitative cytochemistry of glycogen in blood cells. Methods and clinical application.

    PubMed

    Gahrton, G; Yataganas, X

    1976-01-01

    Quantitative glycogen determinations can be made in single blood and bone marrow cells, using microspectrophotometry or microfluorometry after staining with variants of the periodic acid--Schiff (PAS) reaction. These PAS variant reactions generally do not indicate the presence of non-glycogen PAS-positive substances, known to be prevalent in various hematopoietic cells, possibly due to masking of reactive groups. The specificity of the reaction in blood cells was ascertained by alpha-amylase digestion, which removed more than 95% of the PAS-positive material. Calibration of the PAS reaction was undertaken with a microdroplet model of pure leukocyte glycogen. The glycogen amounts in the droplets were determined by microinterferometry, the droplets were stained with a variant PAS reaction, and the total extinction of the reaction product in the stained droplets was determined by microspectrophotometry. The extinction coefficient (k) was obtained from the equation k equals Etot divided by M where (Etot) is the total extinction as determined by microspectrophotometry and (M) the dry glycogen amount as determined by microinterferometry. The microinterferometric dry mass determinations were calibrated by X-ray absorption in order to obtain the absolute amounts of glycogen. For practical purposes a reference system was made of normal neutrophil leukocytes. The glycogen content in the reference neutrophils was first determined with the micromodel. These neutrophils, now with a known glycogen amount, were stained with the PAS reagents and measured microspectrophotometrically in parallel with cells containing an unknown glycogen amount. Alternatively, the staining was made with a fluorescent PAS reaction, and the glycogen content determined by microfluorometry. Both methods appeared suitable for determining the glycogen content of blood cells from patients with various diseases, though the microfluorometric method was preferable for measurements of small amounts of

  11. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 19 Customs Duties 1 2014-04-01 2014-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  12. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  13. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 19 Customs Duties 1 2012-04-01 2012-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  14. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 19 Customs Duties 1 2011-04-01 2011-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  15. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 19 Customs Duties 1 2013-04-01 2013-04-01 false Painted, colored or stained glass windows for.... General Provisions Works of Art § 10.52 Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are claimed free of duty...

  16. A comparative study of quantitative stains for DNA in image cytometry.

    PubMed

    Mikel, U V; Becker, R L

    1991-08-01

    In this study we examined the reproducibility of several stains used to measure nuclear DNA by image cytometry. The specimens were touch preparations of liver and testis from mouse and liver, intestine and brain from rat, fixed in either neutral formalin or Carnoy's solution. The tested stains included four Feulgen methods (pararosaniline, azure-A, thionin and acriflavine), the gallocyanine-chromalum stain and two fluorescent stains (acridine orange and propidium iodide). Absorbance measurements employed a video image analysis system; fluorescence measurements were from a scanning microspectrophotometer. The acriflavine-Feulgen stain was analyzed for both absorbance and fluorescence. All seven stains were quantitative for DNA and gave reproducible results. The absorbance measurements had a lower coefficient of variation (CV) than the fluorescence values. In a nested analysis of variance of the pararosaniline Feulgen stains, cell-to-cell variability accounted for 67% of the total variance; slide-to-slide, 9%; and batch-to-batch, 24%. These values did not change significantly when the staining was performed in an automatic staining machine. For DNA analysis using image cytometry, we conclude that the Feulgen staining technique is the most useful. In particular, acriflavine-Feulgen-stained cells fixed in Carnoy's fluid give the least variation between measurement values and the most accurate ratios between the separate ploidy groups. For fluorescence cytometry we recommend Carnoy's fixation and the acriflavine-Feulgen stain because of its narrow CV as compared to acridine orange and propidium iodide. PMID:1718295

  17. Elimination of iron pigments and background staining which mask immunoperoxidase reactions.

    PubMed

    Itoiz, M E; Orrea, S

    1983-01-01

    When immunohistochemical stainings are applied to demonstration of antigens in histopathological specimens, the ferrous pigments which may be present in the tissues usually mask the final precipitates of the reaction. These pigments can be eliminated with oxalic acid or sodium dithionite after the immunohistochemical staining. These treatments also help in the bleaching of unspecific background stain. PMID:6192670

  18. A Comparison of Heat versus Methanol Fixation for Gram Staining Bacteria

    ERIC Educational Resources Information Center

    Minnerath, Jeanne M.; Roland, Jenna M.; Rossi, Lucas C.; Weishalla, Steven R.; Wolf, Melissa M.

    2009-01-01

    Gram staining bacteria is a fundamental technique introduced in general biology and microbiology laboratory courses. Two common problems students encounter when Gram staining bacteria are (1) having a difficult time locating bacterial cells on the microscope slide and (2) over-decolorizing bacterial cells during the staining procedure such that…

  19. Digital simulation of staining in histopathology multispectral images: enhancement and linear transformation of spectral transmittance.

    PubMed

    Bautista, Pinky A; Yagi, Yukako

    2012-05-01

    Hematoxylin and eosin (H&E) stain is currently the most popular for routine histopathology staining. Special and/or immuno-histochemical (IHC) staining is often requested to further corroborate the initial diagnosis on H&E stained tissue sections. Digital simulation of staining (or digital staining) can be a very valuable tool to produce the desired stained images from the H&E stained tissue sections instantaneously. We present an approach to digital staining of histopathology multispectral images by combining the effects of spectral enhancement and spectral transformation. Spectral enhancement is accomplished by shifting the N-band original spectrum of the multispectral pixel with the weighted difference between the pixel's original and estimated spectrum; the spectrum is estimated using M < N principal component (PC) vectors. The pixel's enhanced spectrum is transformed to the spectral configuration associated to its reaction to a specific stain by utilizing an N × N transformation matrix, which is derived through application of least mean squares method to the enhanced and target spectral transmittance samples of the different tissue components found in the image. Results of our experiments on the digital conversion of an H&E stained multispectral image to its Masson's trichrome stained equivalent show the viability of the method. PMID:22612136

  20. Use of eriochrome cyanine R for routine histology and histopathology: an improved dichromatic staining procedure.

    PubMed

    Stefanović, D

    2015-01-01

    A modified dichromatic iron-eriocyanine R (Fe-ECR) staining method is described. Staining obtained with this new technique generally was similar to that of hematoxylin and eosin (H & E). Cell nuclei were stained blue. Cardiac, smooth and skeletal muscle, and red blood cells, were stained different shades of red. Collagen fibers were stained different shades of orange, usually faintly. Decalcified bony tissue was stained pinkish violet. Epithelial cells were strongly stained deep shades of red, magenta and violet. Cartilage matrix, and goblet and mast cells were unstained. Although Fe-ECR staining differed too much from standard H & E staining to be a substitute for diagnostic purposes, the dichromatic method described might usefully replace van Gieson or trichrome stains, especially if muscle is of interest. A pH 0.95 staining solution was used to differentiate initially over-stained sections followed by washing in distilled water. This dichromatic technique is easier to perform and more precisely controllable than other ECR dichromatic methods. The entire procedure can be completed in less than 5 min. The technique has the advantages of greater technical simplicity and speed, a larger range of polychromasia, and a longer shelf-life than H & E. ECR also is more reliably available than hematoxylin and usually is less expensive. PMID:26140653

  1. Evaluation of an indirect fluorescent-antibody stain for detection of Pneumocystis carinii in respiratory specimens.

    PubMed Central

    Ng, V L; Yajko, D M; McPhaul, L W; Gartner, I; Byford, B; Goodman, C D; Nassos, P S; Sanders, C A; Howes, E L; Leoung, G

    1990-01-01

    Two prospective studies were undertaken to evaluate a commercial indirect fluorescent-antibody (IFA) stain for the detection of Pneumocystis carinii in respiratory specimens from individuals at risk for or with the acquired immunodeficiency syndrome. The first study compared IFA with Diff-Quik (DQ; a rapid Giemsa-like stain) for detecting P. carinii in 95 induced sputa obtained from 77 asymptomatic patients who had survived one previous episode of P. carinii pneumonia and who were being treated prophylactically with aerosolized pentamidine. Only one induced sputum specimen was found to contain P. carinii; organisms were detected by both stains. The second study compared the performance of the IFA stain versus DQ, modified toluidine blue O, and Gomori methenamine silver stains for detecting P. carinii in symptomatic individuals at risk for or with acquired immunodeficiency syndrome. Of 182 specimens examined, P. carinii was detected in 105 by one or more stains; the DQ stain detected 73 (70%), the modified toluidine blue O stain detected 75 (71%), the Gomori methenamine silver stain detected 76 (72%), and the IFA stain detected 95 (90%). The IFA stain was more sensitive (P less than 0.01) than the other traditional stains for detecting P. carinii; however, a subsequent clinical evaluation revealed that a subset of IFA-positive-only specimens were from patients whose clinical symptoms resolved without specific anti-P. carinii therapy. Images PMID:1693631

  2. A study to evaluate the efficacy of xylene-free hematoxylin and eosin staining procedure as compared to the conventional hematoxylin and eosin staining: An experimental study

    PubMed Central

    Ankle, Madhuri R; Joshi, Priya S

    2011-01-01

    Context: Use of diluted dish washing solution (DWS) has been experimented successfully as a substitute for xylene to deparaffinize tissue sections during hematoxylin and eosin (H and E) staining. Aims: (1) Test the hypothesis that xylene- and methanol-free sections (XMF) deparaffinized with diluted DWS are better than or at par with conventional H and E sections. (2) To compare the efficacy of xylene-free sections with the conventional H and E sections. Settings and Design: Single blinded experimental study. Materials and Methods: Sixty paraffin blocks were considered. One section was stained with conventional H and E method (Group A) and the other with XMF H and E (Group B). Slides were scored for parameters; nuclear staining, cytoplasmic staining (adequate = score1, inadequate = score0), uniformity, clarity, crispness (present = score1, absent = score0). Score >/= 2 was inadequate for diagnosis and 3-5 was adequate for diagnosis. Statistical analysis used: Z test. Results: Adequate nuclear staining, 96.66% sections in group A and 98.33% in Group B (Z = 0.59, P>0.05); adequate cytoplasmic staining, 93.33% in group A and 83.33% in Group B (Z = 1.97, P<0.05); uniform staining, 70% in group A, 50% in group B (Z = 1.94, P<0.05), clarity present in 85% of group A, 88.33% of group B sections (Z = 0.27, P>0.05), crisp staining in 76.66% in group A and 83.33% in Group B (Z = 1.98, P<0.05), 88.33% Group A sections stained adequately for diagnosis as compared with 90% in Group B (Z = 0.17, P>0.05). Conclusion: Xylene- and methanol-free H and E staining is a better alternative to the conventional H and E staining procedure. PMID:22529574

  3. Gain-of-function mutations cluster in distinct regions associated with the signalling pathway in the PAS domain of the aerotaxis receptor, Aer.

    PubMed

    Campbell, Asharie J; Watts, Kylie J; Johnson, Mark S; Taylor, Barry L

    2010-08-01

    The Aer receptor monitors internal energy (redox) levels in Escherichia coli with an FAD-containing PAS domain. Here, we randomly mutagenized the region encoding residues 14-119 of the PAS domain and found 72 aerotaxis-defective mutants, 24 of which were gain-of-function, signal-on mutants. The mutations were mapped onto an Aer homology model based on the structure of the PAS-FAD domain in NifL from Azotobacter vinlandii. Signal-on lesions clustered in the FAD binding pocket, the beta-scaffolding and in the N-cap loop. We suggest that the signal-on lesions mimic the 'signal-on' state of the PAS domain, and therefore may be markers for the signal-in and signal-out regions of this domain. We propose that the reduction of FAD rearranges the FAD binding pocket in a way that repositions the beta-scaffolding and the N-cap loop. The resulting conformational changes are likely to be conveyed directly to the HAMP domain, and on to the kinase control module. In support of this hypothesis, we demonstrated disulphide band formation between cysteines substituted at residues N98C or I114C in the PAS beta-scaffold and residue Q248C in the HAMP AS-2 helix. PMID:20545849

  4. A Novel Contrast Stain for the Rapid Diagnosis of Pityriasis Versicolor: A Comparison of Chicago Sky Blue 6B Stain, Potassium Hydroxide Mount and Culture

    PubMed Central

    Lodha, Nikita; Poojary, Shital Amin

    2015-01-01

    Background: The mycological study of pityriasis versicolor is usually done by potassium hydroxide (KOH) mount and culture. However, KOH mount lacks a color contrast and requires a trained eye to interpret, while culture is difficult to perform, time consuming and has low sensitivity. Chicago Sky Blue 6B (CSB) is a new contrast stain that highlights the fungal hyphae and spores, blue against a purplish background. Aims and Objectives: This study was done to compare the utility of a novel contrast stain (CSB stain) with KOH mount and culture. Materials and Methods: Skin scrapings from the lesions of 100 clinically diagnosed cases of P. versicolor were subjected to (1) KOH mount and CSB stain for direct microscopic examination and (2) culture using Sabouraud's dextrose agar. The statistical analysis of CSB stain and culture was done using KOH mount as the reference method, as it is the most commonly performed and practical diagnostic test available for P. versicolor. An interrater reliability analysis using the Cohen's Kappa statistic was performed to determine consistency (agreement) among the different modalities. Observations and Results: Direct microscopy with CSB stain, KOH mount and mycological culture showed positive results in 98 (98%), 92 (92%) and 56 (56%) patients, respectively. Using KOH mount as the reference method, CSB stain had a sensitivity of 100% which was significantly higher than culture (60.9%). Statistically significant fair agreement was found between CSB stain and KOH mount (94% with κ=0.38, P < 0.001). Negligible agreement was found between CSB stain and culture (66%, κ=0.199, P = 0.001) as well as between KOH mount and culture (64%, κ=0.051, P = 0.107). Conclusion: CSB staining of skin scrapings is the most sensitive method for the diagnosis of pityriasis versicolor. Due to the distinct contrast provided by CSB, it is easy to perform, rapid and qualitatively superior to KOH mount. PMID:26288400

  5. Morphological Analysis of Cell Death by Cytospinning Followed by Rapid Staining.

    PubMed

    Crowley, Lisa C; Marfell, Brooke J; Waterhouse, Nigel J

    2016-01-01

    Identifying and characterizing different forms of cell death can be facilitated by staining internal cellular structures with dyes such as hematoxylin and eosin (H&E). These dyes stain the nucleus and cytoplasm, respectively, and optimized reagents (e.g., Rapi-Diff, Rapid Stain, or Quick Dip) are commonly used in pathology laboratories. Fixing and staining adherent cells with these optimized reagents is a straightforward procedure, but apoptotic cells may detach from the culture plate and be washed away during the fixing and staining procedure. To prevent the loss of apoptotic cells, cells can be gently centrifuged onto glass slides by cytospinning before fixing and staining. In addition to apoptotic cells, this procedure can be used on cells in suspension, or adherent cells that have been trypsinized and removed from the culture dish. This protocol describes cytospinning followed by Rapi-Diff staining for morphological analysis of cell death. PMID:27587773

  6. Copper iodide staining and determination of proteins adsorbed to microtiter plates.

    PubMed

    Root, D D; Reisler, E

    1990-04-01

    Copper iodide staining and determination of proteins adsorbed to polystyrene microtiter plates are described. The minimum amount of copper iodide-stained protein detected in densitometric measurements is approximately 20 pg/mm2. Enzyme immunoassay readers may also be used for the determination of copper iodide-stained proteins, but are less sensitive than densitometers. The densitometric readings of copper iodide-stained proteins vary linearly with the amount of protein present as verified by enzymatic and radioactive probes. Staining is complete in 2-3 min and may be removed by a 30-min treatment with EDTA without loss of adsorbed protein or immunoreactivity. The exact amount of protein adsorbed to microtiter plate wells can be measured by using protein bound and stained on nitrocellulose as a calibration curve. Copper iodide staining is a rapid, convenient, and inexpensive alternative to radioactive measurements of similar parameters. PMID:1694063

  7. A clinically motivated 2-fold framework for quantifying and classifying immunohistochemically stained specimens.

    PubMed

    Hall, Bonnie; Chen, Wenjin; Reiss, Michael; Foran, David J

    2007-01-01

    Motivated by the current limitations of automated quantitative image analysis in discriminating among intracellular immunohistochemical (IHC) staining patterns, this paper presents a two-fold approach for IHC characterization that utilizes both the protein stain information and the surrounding tissue architecture. Through the use of a color unmixing algorithm, stained tissue sections are automatically decomposed into the IHC stain, which visualizes the target protein, and the counterstain which provides an objective indication of the underlying histologic architecture. Feature measures are subsequently extracted from both staining planes. In order to characterize the IHC expression pattern, this approach exploits the use of a non-traditional feature based on textons. Novel biologically motivated filter banks are introduced in order to derive texture signatures for different IHC staining patterns. Systematic experiments using this approach were used to classify breast cancer tissue microarrays which had been previously prepared using immuno-targeted nuclear, cytoplasmic, and membrane stains. PMID:18044580

  8. Previsible silver staining of protein in electrophoresis gels with mass spectrometry compatibility.

    PubMed

    Jin, Li-Tai; Li, Xiao-Kun; Cong, Wei-Tao; Hwang, Sun-Young; Choi, Jung-Kap

    2008-12-15

    A convenient silver staining method for protein in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels is described. The method is previsible, sensitive, and mass spectrometry (MS) compatible. Two visible counter ion dyes, ethyl violet (EV) and zincon (ZC), were used in the first staining solution with a detection limit of 2 to 8 ng/band in approximately 1h. The dye-stained gel can be further stained by silver staining, which is based on acidic silver staining employing ZC with sodium thiosulfate as silver ion sensitizers. Especially, ZC has silver ion reducing power by cleavage of the diazo bond of the dye during silver reduction. The second silver staining can be completed in approximately 1h with a detection limit of 0.2 ng/band. PMID:18804088

  9. Using narrow-band J-PAS photometry to assess the properties of the stellar population in galaxies

    NASA Astrophysics Data System (ADS)

    Bruzual, Gustavo; Mejia-Narvaez, Alfredo; Magris C., Gladis

    2015-08-01

    We study the uncertainties and biases on the properties of the stellar population content of galaxies retrieved from narrow-band (J-PAS) photometry using the non-parametric method of spectral fitting dubbed DynBaS. We construct a star formation history library à la Chen et al. (2012), and then SED-fit a selection of synthetic spectra with observational properties similar to SDSS galaxies. We confront the results obtained from the photometric fits to those obtained from spectroscopic data for synthetic and real galaxies at various redshift ranges. Since no assumption on the star formation history is made, the so called template mismatch biases are naturally overcome. We find that biases in our estimations are the consequence of the several degeneracies between mass, age, metallicity, and internal dust extinction present in galaxy properties.

  10. Human mutation within Per-Arnt-Sim (PAS) domain-containing protein kinase (PASK) causes basal insulin hypersecretion.

    PubMed

    Semplici, Francesca; Vaxillaire, Martine; Fogarty, Sarah; Semache, Meriem; Bonnefond, Amélie; Fontés, Ghislaine; Philippe, Julien; Meur, Gargi; Diraison, Frederique; Sessions, Richard B; Rutter, Jared; Poitout, Vincent; Froguel, Philippe; Rutter, Guy A

    2011-12-23

    PAS kinase (PASK) is a glucose-regulated protein kinase involved in the control of pancreatic islet hormone release and insulin sensitivity. We aimed here to identify mutations in the PASK gene that may be associated with young-onset diabetes in humans. We screened 18 diabetic probands with unelucidated maturity-onset diabetes of the young (MODY). We identified two rare nonsynonymous mutations in the PASK gene (p.L1051V and p.G1117E), each of which was found in a single MODY family. Wild type or mutant PASKs were expressed in HEK 293 cells. Kinase activity of the affinity-purified proteins was assayed as autophosphorylation at amino acid Thr307 or against an Ugp1p-derived peptide. Whereas the PASK p.G1117E mutant displayed a ∼25% increase with respect to wild type PASK in the extent of autophosphorylation, and a ∼2-fold increase in kinase activity toward exogenous substrates, the activity of the p.L1051V mutant was unchanged. Amino acid Gly1117 is located in an α helical region opposing the active site of PASK and may elicit either: (a) a conformational change that increases catalytic efficiency or (b) a diminished inhibitory interaction with the PAS domain. Mouse islets were therefore infected with adenoviruses expressing wild type or mutant PASK and the regulation of insulin secretion was examined. PASK p.G1117E-infected islets displayed a 4-fold decrease in glucose-stimulated (16.7 versus 3 mM) insulin secretion, chiefly reflecting a 4.5-fold increase in insulin release at low glucose. In summary, we have characterized a rare mutation (p.G1117E) in the PASK gene from a young-onset diabetes family, which modulates glucose-stimulated insulin secretion. PMID:22065581

  11. Aggrandizing oral submucous fibrosis grading using an adjunct special stain: A pilot study

    PubMed Central

    Reshma, V; Varsha, BK; Rakesh, P; Radhika, MB; Soumya, M; D’Mello, Sarah

    2016-01-01

    Introduction: Oral submucous fibrosis (OSMF) is graded according to various histological factors which include the epithelial changes and the connective tissue changes. These features though could be identified in routine hematoxylin and eosin (H and E) staining; they could be better appreciated in special stains. This pilot study is an attempt to identify a single special stain that can act as an adjunct to H and E stain to help grade this potentially malignant disease. Aims and Objectives: To assess if special stains can improvise on differentiating the various histological changes seen in OSMF and to accordingly grade OSMF cases. Materials and Methods: Formalin-fixed paraffin-embedded tissue sections of OSMF-10 cases of each grade (n = 30). Three special stains: Van-Gieson, Mallory's trichrome and Masson trichrome. Statistical Analysis: The results obtained were tabulated and statistically analyzed using Chi-square test. Observations and Results: The thickness and degree of keratinization were best detected in Mallory's stain (100%) and were statistically significant; the subepithelial changes were better detected using special stains, especially Mallory's stain (100%). The changes in collagen fibers were better visualized in all three special stains but were not statistically significant. The changes in blood vessels were better detected in Van-Gieson's and Mallory's stain; the obtained results were statistically significant. The degree of fibrosis between muscle bundles could be detected in all the three special stains, but when compared the results were not statistically significant. The questionable areas of muscle degeneration, especially in deeper connective tissue were better detected in Mallory's (43%) and Masson's stain (43%) as compared to Van-Gieson stain (14%) and the results obtained were statistically significant. The inflammatory cells and dysplastic features are better visualized in routine H and E stains. Conclusion: Pathogenesis of OSMF is

  12. Novel genetic mutations in a sporadic port-wine stain.

    PubMed

    Lian, Christine Guo; Sholl, Lynette M; Zakka, Labib R; O, Teresa M; Liu, Cynthia; Xu, Shuyun; Stanek, Ewelina; Garcia, Elizabeth; Jia, Yonghui; MacConaill, Laura E; Murphy, George F; Waner, Milton; Mihm, Martin C

    2014-12-01

    IMPORTANCE Port-wine stains (PWSs) are common congenital cutaneous capillary malformations. A somatic GNAQ mutation was recently identified in patients with sporadic PWSs and Sturge-Weber syndrome. However, subsequent studies to confirm or extend this observation are lacking.OBSERVATIONS We report a long-standing, unilateral facial PWS of a man in his early 70s confirmed by histopathological analysis. Staged surgical excision of the vascular malformation was performed, and genomic DNA was extracted from the vascular malformation specimen and normal skin. Targeted next-generation sequencing of the coding sequence of 275 known cancer genes including GNAQ was performed in both specimens. A single-nucleotide variant(c.548G>A, p.Arg183Gln) in GNAQ was identified in the PWS-affected tissue but not in the normal skin sample. In addition, this sequencing approach uncovered several additional novel somatic mutations in the genes SMARCA4, EPHA3, MYB, PDGFR-β, and PIK3CA.CONCLUSIONS AND RELEVANCE Our findings confirm the presence of somatic mutations inGNAQ in the affected skin of a patient with congenital PWS, as well as alterations in several other novel genes of possible importance in the pathogenesis of PWS that may also offer substantial therapeutic targets. PMID:25188413

  13. Diffuse reflectance FTIR of stains on grit blasted metals

    SciTech Connect

    Powell, G.L.; Hallman, R.L. Jr.; Cox, R.L.

    1997-08-09

    Diffuse reflectance mid-infrared Fourier transform (DRIFT) spectroscopy has been applied to the detection of oil contamination on grit-blasted metals. The object of this application is to detect and discriminate between silicone and hydrocarbon oil contamination at levels approaching 10 mg/m{sup 2}. A portable FTIR spectrometer with dedicated diffuse reflectance optics was developed for this purpose. Using translation devices positioned by instructions from the spectrometer operating system, images of macroscopic substrates were produced with millimeter spatial resolution. The pixels that comprise an image are each a full mid-infrared spectrum with excellent signal-to-noise, each determined as individual files and uniquely saved to disc. Reduced spectra amplitudes, based on peak height, area, or other chemometric techniques, mapped as a function of the spatial coordinates of the pixel are used to display the image. This paper demonstrates the application of the technique to the analysis of stains on grit-blasted metals, including the calibration of the method, the inspection of substrates, and the migration of oil contamination.

  14. Development of a stained cell nuclei counting system

    NASA Astrophysics Data System (ADS)

    Timilsina, Niranjan; Moffatt, Christopher; Okada, Kazunori

    2011-03-01

    This paper presents a novel cell counting system which exploits the Fast Radial Symmetry Transformation (FRST) algorithm [1]. The driving force behind our system is a research on neurogenesis in the intact nervous system of Manduca Sexta or the Tobacco Hornworm, which was being studied to assess the impact of age, food and environment on neurogenesis. The varying thickness of the intact nervous system in this species often yields images with inhomogeneous background and inconsistencies such as varying illumination, variable contrast, and irregular cell size. For automated counting, such inhomogeneity and inconsistencies must be addressed, which no existing work has done successfully. Thus, our goal is to devise a new cell counting algorithm for the images with non-uniform background. Our solution adapts FRST: a computer vision algorithm which is designed to detect points of interest on circular regions such as human eyes. This algorithm enhances the occurrences of the stained-cell nuclei in 2D digital images and negates the problems caused by their inhomogeneity. Besides FRST, our algorithm employs standard image processing methods, such as mathematical morphology and connected component analysis. We have evaluated the developed cell counting system with fourteen digital images of Tobacco Hornworm's nervous system collected for this study with ground-truth cell counts by biology experts. Experimental results show that our system has a minimum error of 1.41% and mean error of 16.68% which is at least forty-four percent better than the algorithm without FRST.

  15. The challenges of analysing blood stains with hyperspectral imaging

    NASA Astrophysics Data System (ADS)

    Kuula, J.; Puupponen, H.-H.; Rinta, H.; Pölönen, I.

    2014-06-01

    Hyperspectral imaging is a potential noninvasive technology for detecting, separating and identifying various substances. In the forensic and military medicine and other CBRNE related use it could be a potential method for analyzing blood and for scanning other human based fluids. For example, it would be valuable to easily detect whether some traces of blood are from one or more persons or if there are some irrelevant substances or anomalies in the blood. This article represents an experiment of separating four persons' blood stains on a white cotton fabric with a SWIR hyperspectral camera and FT-NIR spectrometer. Each tested sample includes standardized 75 _l of 100 % blood. The results suggest that on the basis of the amount of erythrocytes in the blood, different people's blood might be separable by hyperspectral analysis. And, referring to the indication given by erythrocytes, there might be a possibility to find some other traces in the blood as well. However, these assumptions need to be verified with wider tests, as the number of samples in the study was small. According to the study there also seems to be several biological, chemical and physical factors which affect alone and together on the hyperspectral analyzing results of blood on fabric textures, and these factors need to be considered before making any further conclusions on the analysis of blood on various materials.

  16. The Application of Molecular Diagnostics to Stained Cytology Smears.

    PubMed

    Oktay, Maja H; Adler, Esther; Hakima, Laleh; Grunblatt, Eli; Pieri, Evan; Seymour, Andrew; Khader, Samer; Cajigas, Antonio; Suhrland, Mark; Goswami, Sumanta

    2016-05-01

    Detection of mutational alterations is important for guiding treatment decisions of lung non-small-cell carcinomas and thyroid nodules with atypical cytologic findings. Inoperable lung tumors requiring further testing for staging and thyroid lesions often are diagnosed using only cytology material. Molecular diagnostic tests of these samples typically are performed on cell blocks; however, insufficient cellularity of cell blocks is a limitation for test performance. In addition, some of the fixatives used while preparing cell blocks often introduces artifacts for mutation detection. Here, we applied qClamp xenonucleic technology and quantitative RT-PCR to cells microdissected directly from stained cytology smears to detect common alterations including mutations and translocations in non-small-cell carcinomas and thyroid lesions. By using this approach, we achieved a 1% molecular alteration detection rate from as few as 50 cells. Ultrasensitive methods of molecular alteration detection similar to the one described here will be increasingly important for the evaluation of molecular alterations in clinical scenarios when only tissue samples that are small are available. PMID:26921541

  17. An automatic nonrigid registration for stained histological sections.

    PubMed

    Auer, Martin; Regitnig, Peter; Holzapfel, Gerhard A

    2005-04-01

    Automatic computer-based analyses of histological sections which are differently stained require that they are related to each other. Most registration methods are only able to perform rigid-body motion and are sensitive to noise and artifacts. Histological images, however, are accompanied by several artifacts and different contrasts, which require a nonrigid registration. In this paper, we present a hierarchical nonrigid registration algorithm able to align images, which contain minor image artifacts. The algorithm requires no a priori knowledge of the true image. The hierarchical design of the algorithm enhances robustness and accuracy, and saves computational costs. The proposed algorithm is decomposed into a fast, coarse, rigid registration step and a slower, but finer, nonrigid step. For the coarse registration, we use image pyramids, while for the second step, we combine a point-based registration with an elastic thin-plate spline interpolation. Accuracy tests, performed for 20 histological images obtained from human arteries, have shown that the error measure is acceptable, and that the image noise does not cause a problem. The associated convergence rate of the mean pixel displacement error during the rigid and nonrigid registrations is satisfying. The algorithm can be applied to various multicontrast elastic registration problems in medical imaging and may be extended to three dimensions. PMID:15825482

  18. Histochemical staining of orbicularis oculi muscle in ectropion and entropion.

    PubMed

    Manners, R M; Weller, R O

    1994-01-01

    A histochemical study of orbicularis oculi was undertaken to test the hypothesis that there is a difference in the percentage and size of muscle fibre types which accounts for the development of involutional ectropion or entropion. Wedge excisions from lower lids of patients undergoing repair of these conditions were frozen-sectioned and stained histochemically to reveal muscle fibre types. Five ectropion and five entropion specimens were obtained, and the percentage of type 1 and type 2 fibres, fibre perimeters and fibre diameters were measured. An abundance of type 2 fibres was found in both ectropion (mean 89.6%) and entropion (mean 82.6%). No significant difference was found with respect to fibre type, perimeter or diameter when ectropion was compared with entropion or when either was compared with normals. Type 2 fibres were larger than type 1 in both ectropion and entropion. We conclude that no significant difference could be identified between orbicularis muscle fibres in ectropion, entropion and normals to account for the development of the eyelid malpositions. PMID:7958041

  19. H2O2 production rate in Lactobacillus johnsonii is modulated via the interplay of a heterodimeric flavin oxidoreductase with a soluble 28 Kd PAS domain containing protein

    PubMed Central

    Valladares, Ricardo B.; Graves, Christina; Wright, Kaitlyn; Gardner, Christopher L.; Lorca, Graciela L.; Gonzalez, Claudio F.

    2015-01-01

    Host and commensals crosstalk, mediated by reactive oxygen species (ROS), has triggered a growing scientific interest to understand the mechanisms governing such interaction. However, the majority of the scientific studies published do not evaluate the ROS production by commensals bacteria. In this context we recently showed that Lactobacillus johnsonii N6.2, a strain of probiotic value, modulates the activity of the critical enzymes 2,3-indoleamine dioxygenase via H2O2 production. L. johnsonii N6.2 by decreasing IDO activity, is able to modify the tryptophan/kynurenine ratio in the host blood with further systemic consequences. Understanding the mechanisms of H2O2 production is critical to predict the probiotic value of these strains and to optimize bacterial biomass production in industrial processes. We performed a transcriptome analysis to identify genes differentially expressed in L. johnsonii N6.2 cells collected from cultures grown under different aeration conditions. Herein we described the biochemical characteristics of a heterodimeric FMN reductase (FRedA/B) whose in vitro activity is controlled by LjPAS protein with a typical Per-Arnst-Sim (PAS) sensor domain. Interestingly, LjPAS is fused to the FMN reductase domains in other lactobacillaceae. In L. johnsonii, LjPAS is encoded by an independent gene which expression is repressed under anaerobic conditions (>3 fold). Purified LjPAS was able to slow down the FRedA/B initial activity rate when the holoenzyme precursors (FredA, FredB, and FMN) were mixed in vitro. Altogether the results obtained suggest that LjPAS module regulates the H2O2 production helping the cells to minimize oxidative stress in response to environmental conditions. PMID:26236298

  20. Deciphering Dimerization Modes of PAS Domains: Computational and Experimental Analyses of the AhR:ARNT Complex Reveal New Insights Into the Mechanisms of AhR Transformation

    PubMed Central

    Corrada, Dario; Soshilov, Anatoly A.; Denison, Michael S.

    2016-01-01

    The Aryl hydrocarbon Receptor (AhR) is a transcription factor that mediates the biochemical response to xenobiotics and the toxic effects of a number of environmental contaminants, including dioxins. Recently, endogenous regulatory roles for the AhR in normal physiology and development have also been reported, thus extending the interest in understanding its molecular mechanisms of activation. Since dimerization with the AhR Nuclear Translocator (ARNT) protein, occurring through the Helix-Loop-Helix (HLH) and PER-ARNT-SIM (PAS) domains, is needed to convert the AhR into its transcriptionally active form, deciphering the AhR:ARNT dimerization mode would provide insights into the mechanisms of AhR transformation. Here we present homology models of the murine AhR:ARNT PAS domain dimer developed using recently available X-ray structures of other bHLH-PAS protein dimers. Due to the different reciprocal orientation and interaction surfaces in the different template dimers, two alternative models were developed for both the PAS-A and PAS-B dimers and they were characterized by combining a number of computational evaluations. Both well-established hot spot prediction methods and new approaches to analyze individual residue and residue-pairwise contributions to the MM-GBSA binding free energies were adopted to predict residues critical for dimer stabilization. On this basis, a mutagenesis strategy for both the murine AhR and ARNT proteins was designed and ligand-dependent DNA binding ability of the AhR:ARNT heterodimer mutants was evaluated. While functional analysis disfavored the HIF2α:ARNT heterodimer-based PAS-B model, most mutants derived from the CLOCK:BMAL1-based AhR:ARNT dimer models of both the PAS-A and the PAS-B dramatically decreased the levels of DNA binding, suggesting this latter model as the most suitable for describing AhR:ARNT dimerization. These novel results open new research directions focused at elucidating basic molecular mechanisms underlying the

  1. Analysis of the Microbiota of Black Stain in the Primary Dentition

    PubMed Central

    Li, Yue; Zhang, Qian; Zhang, Fangfei; Liu, Ruoxi; Liu, He; Chen, Feng

    2015-01-01

    Black tooth stain is a characteristic extrinsic discoloration commonly seen on the cervical enamel following the contour of the gingiva. To investigate the relationship between black tooth stain and the oral microbiota, we used 16S rRNA gene sequencing to compare the microbial composition of dental plaque and saliva among caries-free children with and without black stain. Dental plaque and saliva, as well as black stain, were sampled from 10 children with and 15 children without black stain. Data were analyzed using the pipeline tool MOTHUR. Student’s t-test was used to compare alpha diversities and the Mann-Whitney U test to compare the relative abundances of the microbial taxa. A total of 10 phyla, 19 classes, 32 orders, 61 families and 102 genera were detected in these samples. Shannon and Simpson diversity were found to be significantly lower in saliva samples of children with black stain. Microbial diversity was reduced in the black stain compared to the plaque samples. Actinomyces, Cardiobacterium, Haemophilus, Corynebacterium, Tannerella and Treponema were more abundant and Campylobacter less abundant in plaque samples of children with black stain. Principal component analysis demonstrated clustering among the dental plaque samples from the control group, while the plaque samples from the black stain group were not and appeared to cluster into two subgroups. Alterations in oral microbiota may be associated with the formation of black stain. PMID:26340752

  2. Detection of infection or infectious agents by use of cytologic and histologic stains.

    PubMed Central

    Woods, G L; Walker, D H

    1996-01-01

    A wide variety of stains are useful for detection of different organisms or, for viruses, the cytopathologic changes they induce, in smears prepared directly from clinical specimens and in tissue sections. Other types of stains, such as hematoxylin and eosin, are used routinely to stain tissue sections and are most valuable for assessing the immunologic response of the host to the invading pathogen. In many cases, the pattern of inflammation provides important clues to diagnosis and helps to guide the selection of additional "special" stains used predominantly for diagnosis of infectious diseases. A stain may be nonspecific, allowing detection of a spectrum of organisms, as do the Papanicolaou stain and silver impregnation methods, or detection of only a limited group of organisms, as do the different acid-fast techniques. Some nonspecific stains, such as the Gram stain, are differential and provide valuable preliminary information concerning identification. Immunohistochemical stains, on the other hand, are specific for a particular organism, although in some cases cross-reactions with other organisms occur. Despite the wealth of information that can be gleaned from a stained smear or section of tissue, however, the specific etiology of an infection often cannot be determined on the basis of only the morphology of the organisms seen; culture data are essential and must be considered in the final diagnosis. PMID:8809467

  3. Analysis of the Microbiota of Black Stain in the Primary Dentition.

    PubMed

    Li, Yue; Zhang, Qian; Zhang, Fangfei; Liu, Ruoxi; Liu, He; Chen, Feng

    2015-01-01

    Black tooth stain is a characteristic extrinsic discoloration commonly seen on the cervical enamel following the contour of the gingiva. To investigate the relationship between black tooth stain and the oral microbiota, we used 16S rRNA gene sequencing to compare the microbial composition of dental plaque and saliva among caries-free children with and without black stain. Dental plaque and saliva, as well as black stain, were sampled from 10 children with and 15 children without black stain. Data were analyzed using the pipeline tool MOTHUR. Student's t-test was used to compare alpha diversities and the Mann-Whitney U test to compare the relative abundances of the microbial taxa. A total of 10 phyla, 19 classes, 32 orders, 61 families and 102 genera were detected in these samples. Shannon and Simpson diversity were found to be significantly lower in saliva samples of children with black stain. Microbial diversity was reduced in the black stain compared to the plaque samples. Actinomyces, Cardiobacterium, Haemophilus, Corynebacterium, Tannerella and Treponema were more abundant and Campylobacter less abundant in plaque samples of children with black stain. Principal component analysis demonstrated clustering among the dental plaque samples from the control group, while the plaque samples from the black stain group were not and appeared to cluster into two subgroups. Alterations in oral microbiota may be associated with the formation of black stain. PMID:26340752

  4. Hyperspectral imaging of the crime scene for detection and identification of blood stains

    NASA Astrophysics Data System (ADS)

    Edelman, G. J.; van Leeuwen, T. G.; Aalders, M. C. G.

    2013-05-01

    Blood stains are an important source of information in forensic investigations. Extraction of DNA may lead to the identification of victims or suspects, while the blood stain pattern may reveal useful information for the reconstruction of a crime. Consequently, techniques for the detection and identification of blood stains are ideally non-destructive in order not to hamper both DNA and the blood stain pattern analysis. Currently, forensic investigators mainly detect and identify blood stains using chemical or optical methods, which are often either destructive or subject to human interpretation. We demonstrated the feasibility of hyperspectral imaging of the crime scene to detect and identify blood stains remotely. Blood stains outside the human body comprise the main chromophores oxy-hemoglobin, methemoglobin and hemichrome. Consequently, the reflectance spectra of blood stains are influenced by the composite of the optical properties of the individual chromophores and the substrate. Using the coefficient of determination between a non-linear least squares multi-component fit and the measured spectra blood stains were successfully distinguished from other substances visually resembling blood (e.g. ketchup, red wine and lip stick) with a sensitivity of 100 % and a specificity of 85 %. The practical applicability of this technique was demonstrated at a mock crime scene, where blood stains were successfully identified automatically.

  5. Staining of in vivo subsurface degradation in dental composites with silver nitrate

    SciTech Connect

    Mair, L.H. )

    1991-03-01

    A previously reported technique for staining areas of degradation in dental composite restorations was evaluated in 51 removed restorations. The staining reagent was silver nitrate, which penetrated the degraded subsurface as ionic silver and was subsequently developed into colored deposits of metallic silver. Several artefacts were recognized that resulted in an apparent image of subsurface stain. Most importantly, the presence of a layer of adsorbed silver on the edge of the specimen exaggerated the extent of staining. In order for the true depth of stain to be determined, thin sections of the materials should first be examined with a stereomicroscope to distinguish any contribution from adsorbed silver on the specimen edge. With this regimen, no stain was present in 41% of the restorations, and in a further 30%, the depth of stain was less than 50 microns. In two composites, the depth of stain was greater than 900 microns, and in a number of specimens, localized stain was found in association with attrition scars. Energy-dispersive x-ray analysis indicated that the amount of silver present in the degraded layers was very small. Overall, the results indicated that the staining technique is useful in the study of composite degradation.

  6. High contrast and homogeneous staining of paraffin sections of whole human brains for three dimensional ultrahigh resolution image analysis.

    PubMed

    Schmitt, O; Eggers, R

    1998-01-01

    The gallocyanin chromalum stain belongs to the classical DNA-RNA staining techniques in histochemistry. It has some important features for successful object orientated image analysis of whole sections of the human brain. To obtain reproducible staining results with these large sections, the method of Einarson was adapted to image analytical requirements. We discuss staining in a warm staining solution, pH adjustment, and optimal stain composition. The embedding procedure for whole human brains is considered as well. PMID:9554583

  7. Digital separation of diaminobenzidine-stained tissues via an automatic color-filtering for immunohistochemical quantification.

    PubMed

    Fu, Rong; Ma, Xiaomian; Bian, Zhaoying; Ma, Jianhua

    2015-02-01

    The digital separation of diaminobenzidine (DAB)-stained tissues from hematoxylin background is an important pre-processing step to analyze immunostains. In most stain separation methods, specific color channels (for example: RGB, HSI, CMYK) or color deconvolution matrices are used to obtain different tissue contrasts between DAB- and hematoxylin-stained areas. However, these methods could produce incomplete separation or color changes because the color spectra of stains and co-localized stains overlap in histological images. Therefore, we proposed an automatic color-filtering to separate hematoxylin- and DAB-stained tissues. In implantation, the RGB images of DAB-labeled immunostains are first converted to 8-bit BN images by a mathematical translation to produce the largest contrast between brown DAB-stained tissues and blue hematoxylin-stained tissues. The first valley in the histogram revised by nonuniform quantization is set as the cut-off point to obtain a brown filter. DAB-stained tissues are accurately delineated from the background counterstain, resulting in DAB-only-image and De-DAB-image. Subsequently, a blue filter is designed in the CIE-Lab color space to further delineate the hematoxylin-stained tissues from the De-DAB-image. Finally, the average values of the remaining pixels of the De-DAB-image are set as the background color of the DAB-only-image to manage uneven dyeing and provide DAB-stained-image for adaptive immunohistochemistry quantitation. Extensive experimental results demonstrated that the proposed method has significant advantages compared with existing methods in terms of complete stain separation without changing the color in DAB-stained areas. PMID:25780744

  8. COMPARISON OF PERMANENT STAINING METHODS FOR THE LABORATORY DIAGNOSIS OF TRICHOMONIASIS

    PubMed Central

    MENEZES, Camila Braz; MELLO, Mariana dos Santos; TASCA, Tiana

    2016-01-01

    Trichomonas vaginalis is the etiologic agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in the world. The diagnosis is based on wet mount preparation and direct microscopy on fixed and stained clinical specimens. The aim of this study was to compare the performance of different fixing and staining techniques used in the detection of T. vaginalis in urine. The smears were fixed and submitted to different methods of permanent staining and then, the morphological aspects of the parasites were analyzed and compared. The Papanicolaou staining with ethanol as the fixative solution showed to be the best method of permanent staining. Our data suggest that staining techniques in association with wet mount examination of fresh specimens contribute to increase the sensitivity in the diagnosis of trichomoniasis. PMID:26910452

  9. Feulgen type staining with Hoffmann's violet-SO2 under exposure to UV rays.

    PubMed

    Dutt, M K

    1979-07-01

    The paper contains an account of the use of Hoffmann's violet-SO2 under exposure to UV rays during staining acid-hydrolysed DNA of mammalian tissue nuclei. Preparations stained with Hoffmann's violet-SO2 without exposure to UV rays reveal extremely pale violet nuclei but when stained under the influence of UV rays show a considerably faster reaction resulting in a very much deeper staining of the nuclei. Sections after staining with this dye-reagent require n-butanol as differentiating reagent. Possible interpretation for the increase in staining ability of this dye-reagent under exposure to UV rays has been elucidated and the reason for considering the reaction as Feulgen type has been discussed. PMID:91084

  10. Understanding Interactions between Cellular Matrices and Metal Complexes: Methods To Improve Silver Nanodot-Specific Staining.

    PubMed

    Choi, Sungmoon; Yu, Junhua

    2016-08-26

    Metal complexes are frequently used for biological applications due to their special photophysical and chemical characteristics. Due to strong interactions between metals and biomacromolecules, a random staining of cytoplasm or nucleoplasm by the complexes results in a low signal-to-background ratio. In this study, we used luminescent silver nanodots as a model to investigate the major driving force for non-specific staining in cellular matrices. Even though some silver nanodot emitters exhibited excellent specific staining of nucleoli, labeling with nanodots was problematic owing to severe non-specific staining. Binding between silver and sulfhydryl group of proteins appeared to be the major factor that enforced the silver staining. The oxidation of thiol groups in cells with hexacyanoferrate(III) dramatically weakened the silver-cell interaction and consequently significantly improved the efficiency of targeted staining. PMID:27380586

  11. Analysis of Formation of Pad Stains in Copper Chemical Mechanical Planarization

    NASA Astrophysics Data System (ADS)

    Lee, Hyosang; Borucki, Leonard; Zhuang, Yun; Joh, Sooyun; O'Moore, Fergal; Philipossian, Ara

    2009-12-01

    A stain model was developed to simulate stain formation on the pad surface in copper chemical and mechanical planarization (CMP). The model consisted of the incompressible Navier-Stokes equations, the heat equation with advection, material removal rate model, a model for generation, transport and deposition of the polishing by-product that produces the stain. Slurry velocity simulations showed shear flow on the land areas and wafer-driven circulation in the grooves. The simulated temperature on the pad and the wafer surface increased gradually in the radial direction; furthermore, temperature simulations showed a 12 °C rise in the reaction temperature on the copper wafer surface. The simulated pad stains deposited on the copper land areas were darker in the direction of wafer rotation, suggesting that the generated staining agents were advected downstream by the slurry flow and deposited on the pad surface in the direction of the wafer rotation. Simulated stain images were in qualitative agreement with experimental results.

  12. Standardization of stain used for diagnosing erythrocytic inclusion body syndrome (EIBS)

    USGS Publications Warehouse

    1987-01-01

    Erythrocytic inclusion body syndrome (EIBS), a viral erythrocytic necrosis (VEN)-like disease, has been observed in several areas in the Northwest. This virus disease is clinically diagnosed by microscopic examination of blood smears for intracytoplasmic erythrocytic inclusion bodies. Fish biologists involved in EIBS diagnostic work have been using several types of hematological stains. It became apparent that standardization of the staining procedure was needed. Comparative tests were conducted on blood smears and kidney imprints with the following commonly used blood stains: (1) Leishman-Giesma, (2) Pinacyanol chloride, (3) Powell 's Giemsa, (4) Harleco's Giemsa, (5) Diff Quik differential stain, (6) Wright's.Pinacyanol chloride stain was found to be the most consistent. The following staining procedure is recommended.

  13. Specimen block counter-staining for localization of GUS expression in transgenic arabidopsis and tobacco

    NASA Technical Reports Server (NTRS)

    Kim, M. K.; Choi, J-W; Jeon, J-H; Franceschi, V. R.; Davin, L. B.; Lewis, N. G.

    2002-01-01

    A simple counter-staining procedure has been developed for comparative beta-glucuronidase (GUS) expression and anatomical localization in transgenic herbaceous arabidopsis and tobacco. This protocol provides good anatomical visualization for monitoring chimeric gene expression at both the organ and tissue levels. It can be used with different histochemical stains and can be extended to the study of woody species. The specimens are paraffin-embedded, the block is trimmed to reveal internal structure, safranin-O staining solution is briefly applied to the surface of the block, then washed off and, after drying, a drop of immersion oil is placed on the stained surface for subsequent photographic work. This gives tissue counter-staining with good structural preservation without loss of GUS staining product; moreover, sample observation is rapid and efficient compared to existing procedures.

  14. Chemical enhancement of footwear impressions in blood on fabric - part 1: protein stains.

    PubMed

    Farrugia, Kevin J; Savage, Kathleen A; Bandey, Helen; Nic Daéid, Niamh

    2011-09-01

    A range of protein stains were utilised for the enhancement of footwear impressions on a variety of fabric types of different colours with blood as a contaminant. A semi-automated stamping device was used to deliver test impressions at a set force to minimise the variability between impressions; multiple impressions were produced and enhanced by each reagent to determine the repeatability of the enhancement. Results indicated that while most protein stains used in this study successfully enhanced impressions in blood on light coloured fabrics, background staining caused interference on natural fabrics. Enhancement on dark coloured fabrics was only achieved using fluorescent protein stains, as non-fluorescent protein stains provided poor contrast. A further comparison was performed with commercially available protein staining solutions and solutions prepared within the laboratory from the appropriate chemicals. Both solutions performed equally well, though it is recommended to use freshly prepared solutions whenever possible. PMID:21889106

  15. Unsupervised color normalisation for H and E stained histopathology image analysis

    NASA Astrophysics Data System (ADS)

    Celis, Raúl; Romero, Eduardo

    2015-12-01

    In histology, each dye component attempts to specifically characterise different microscopic structures. In the case of the Hematoxylin-Eosin (H&E) stain, universally used for routine examination, quantitative analysis may often require the inspection of different morphological signatures related mainly to nuclei patterns, but also to stroma distribution. Nevertheless, computer systems for automatic diagnosis are often fraught by color variations ranging from the capturing device to the laboratory specific staining protocol and stains. This paper presents a novel colour normalisation method for H&E stained histopathology images. This method is based upon the opponent process theory and blindly estimates the best color basis for the Hematoxylin and Eosin stains without relying on prior knowledge. Stain Normalisation and Color Separation are transversal to any Framework of Histopathology Image Analysis.

  16. Role of mucus in gastric mucosal injury induced by local ischemia/reperfusion.

    PubMed

    Seno, K; Joh, T; Yokoyama, Y; Itoh, M

    1995-09-01

    The role of gastric mucus was evaluated in a rat model of gastric epithelial damage induced by local ischemia/reperfusion (I/R) stress. In this model, blood-to-lumen chromium 51-labeled ethylenediaminetetraacetic acid (51Cr-EDTA) clearance served as an index of injury. Tetraprenyl acetone (TPA; 100 mg, 200 mg/kg IP) was used to stimulate mucus production. Administration of TPA increased both the hexosamine content in gastric tissue and the amount of alcian blue-periodic acid Schiff (AB-PAS) stained mucus in the mucosa in a dose-dependent manner. Increases in 51Cr-EDTA clearance induced by I/R were significantly attenuated by TPA in a dose-dependent manner. N-acetyl-L-cysteine (NAC; 0.6%, 0.8%) was perfused into the gastric lumen to assess the effect of reduction in mucus on the injury induced by I/R. Although mean values of hexosamine content were increased by perfusion with NAC, AB-PAS-stained mucus in the mucosa was significantly decreased in a dose-dependent manner. Perfusion of NAC did not change basal 51Cr-EDTA clearance but significantly exacerbated the increase in clearance induced by I/R in a dose-dependent manner. These results indicate that gastric mucus protects the gastric mucosa against I/R stress in vivo. PMID:7665977

  17. Neuropathology in respiratory-related motoneurons in young Pompe (Gaa−/−) mice

    PubMed Central

    Turner, Sara M.F.; Hoyt, Aaron K.; ElMallah, Mai K.; Falk, Darin J.; Byrne, Barry J.; Fuller, David D.

    2016-01-01

    Respiratory and/or lingual dysfunction are among the first motor symptoms in Pompe disease, a disorder resulting from absence or dysfunction of the lysosomal enzyme acid α-glucosidase (GAA). Here, we histologically evaluated the medulla, cervical and thoracic spinal cords in 6 weeks old asymptomatic Pompe (Gaa−/−) mice to determine if neuropathology in respiratory motor regions has an early onset. Periodic acid-Schiff (PAS) staining indicated glycogen accumulation was exclusively occurring in Gaa−/− hypoglossal, mid-cervical and upper thoracic motoneurons. Markers of DNA damage (Tunel) and ongoing apoptosis (Cleaved Caspase 3) did not co-localize with PAS staining, but were prominent in a medullary region which included the nucleus tractus solitarius, and also in the thoracic spinal dorsal horn. We conclude that respiratory-related motoneurons are particularly susceptible to GAA deficiency and that neuronal glycogen accumulation and neurodegeneration may occur independently in early stage disease. The data support early therapeutic intervention in Pompe disease. PMID:26921786

  18. The effect of pulp and seed extract of Citrullus Colocynthis, as an antidaibetic medicinal herb, on hepatocytes glycogen stores in diabetic rabbits

    PubMed Central

    Shafaei, Hajar; Rad, Jafar Soleimani; Delazar, Abbas; Behjati, Mohaddeseh

    2014-01-01

    Background: Medicinal herbs such as Citrullus Colocynthis (C.C) have been used traditionally in the treatment of diabetes mellitus. However therapeutic applications and adverse effects of C.C and its natural variants are not determined well. The current work investigates the effects of pulp and seed extract of C.C on hepatocyte's glycogen stores. Materials and Methods: Thirty six male rabbits were divided into six groups (control and diabetic) randomly. Alloxan was used in order to induce diabetes mellitus in animals. Among 5 diabetic groups, one remained as control and the rest received 100 and 200 mg/kg/day of either pulp or seed extract. One month later, animals were sacrificed and their liver specimen fixed in 10% Formalin was stained with periodic acid schiff (PAS) for light microscopic scanning. Results: PAS staining of hepatocytes revealed large amounts of glycogen stores in diabetic animals treated with pulp and seed extracts of C.C, contrary with non-treated diabetic rabbits. Sites of glycogen deposition were also different in animals treated with seed extract (P < 0.0001). No hepatic congestion was seen in treated animals. Dose escalation has no effect on the obtained results. Conclusions: The anti-diabetic effects of C.C can be explained by its effects on accumulation of glycogen stores in hepatocytes. The importance of varied sites of glycogen deposition by the application of C.C needs to be determined. PMID:25625097

  19. Endocarditis Due to Rare and Fastidious Bacteria

    PubMed Central

    Brouqui, P.; Raoult, D.

    2001-01-01

    The etiologic diagnosis of infective endocarditis is easily made in the presence of continuous bacteremia with gram-positive cocci. However, the blood culture may contain a bacterium rarely associated with endocarditis, such as Lactobacillus spp., Klebsiella spp., or nontoxigenic Corynebacterium, Salmonella, Gemella, Campylobacter, Aeromonas, Yersinia, Nocardia, Pasteurella, Listeria, or Erysipelothrix spp., that requires further investigation to establish the relationship with endocarditis, or the blood culture may be uninformative despite a supportive clinical evaluation. In the latter case, the etiologic agents are either fastidious extracellular or intracellular bacteria. Fastidious extracellular bacteria such as Abiotrophia, HACEK group bacteria, Clostridium, Brucella, Legionella, Mycobacterium, and Bartonella spp. need supplemented media, prolonged incubation time, and special culture conditions. Intracellular bacteria such as Coxiella burnetii cannot be isolated routinely. The two most prevalent etiologic agents of culture-negative endocarditis are C. burnetti and Bartonella spp. Their diagnosis is usually carried out serologically. A systemic pathologic examination of excised heart valves including periodic acid-Schiff (PAS) staining and molecular methods has allowed the identification of Whipple's bacillus endocarditis. Pathologic examination of the valve using special staining, such as Warthin-Starry, Gimenez, and PAS, and broad-spectrum PCR should be performed systematically when no etiologic diagnosis is evident through routine laboratory evaluation. PMID:11148009

  20. Karyotype analysis of four jewel-beetle species (Coleoptera, Buprestidae) detected by standard staining, C-banding, AgNOR-banding and CMA3/DAPI staining

    PubMed Central

    Karagyan, Gayane; Lachowska, Dorota; Kalashian, Mark

    2012-01-01

    Abstract The male karyotypes of Acmaeodera pilosellae persica Mannerheim, 1837 with 2n=20 (18+neoXY), Sphenoptera scovitzii Faldermann, 1835 (2n=38–46), Dicerca aenea validiuscula Semenov, 1895 – 2n=20 (18+Xyp) and Sphaerobothris aghababiani Volkovitsh et Kalashian, 1998 – 2n=16 (14+Xyp) were studied using conventional staining and different chromosome banding techniques: C-banding, AgNOR-banding, as well as fluorochrome Chromomycin A3 (CMA3) and DAPI. It is shown that C-positive segments are weakly visible in all four species which indicates a small amount of constitutive heterochromatin (CH). There were no signals after DAPI staining and some positive signals were discovered using CMA3 staining demonstrating absence of AT-rich DNA and presence of GC-rich clusters of CH. Nucleolus organizing regions (NORs) were revealed using Ag-NOR technique; argentophilic material mostly coincides with positive signals obtained using CMA3 staining. PMID:24260661

  1. Morphometry of cupromeronic blue-stained proteoglycan molecules in animal corneas, versus that of purified proteoglycans stained in vitro, implies that tertiary structures contribute to corneal ultrastructure.

    PubMed Central

    Scott, J E

    1992-01-01

    Isolated, purified small chondroitin (dermatan) sulphate proteoglycans from corneas of cow and rabbit and cow sclera were stained with Cupromeronic blue in 'model' experiments. The lengths and thicknesses of the images were compared with those of the same proteoglycans stained in the tissue, using the critical electrolyte concentration principle to give specificity for sulphated proteoglycans, and keratanase 1 or chondroitinase ABC digestion to distinguish between chondroitin and keratan sulphate. Corrections for orientation of the stained glycan filaments within the section plane were made to convert the observed lengths to true average lengths. Observed lengths of stained chondroitin (dermatan) sulphate were greater than those of keratan sulphate, both in models and tissues, in agreement with published data from biochemical and rotary-shadowing studies, in both species. Corrected (true) average lengths of stained isolated chondroitin (dermatan) sulphate proteoglycans were slightly, but not significantly, longer than expected from rotary shadowing or biochemical measurements. Keratan sulphate lengths were similarly somewhat longer. The data support the idea that Cupromeronic blue acts as a scaffold that helps maintain polyanion shape against distortion on staining. Stained filaments in tissues were sometimes over twice the length of isolated stained proteoglycans, suggesting that 2 glycan chains were aligned end-to-end. Thicknesses of proteoglycan filaments suggested that at least 2 glycan chains were aligned side-by-side, both in models and in tissues. A scheme for proteoglycan tertiary structure in cornea is proposed, in which glycan chains may bridge collagen fibrils in duplexed forms similar to those observed in rotary shadowed preparations. Images Fig. 1 Fig. 2 PMID:1452471

  2. Extinction, seeing and sky transparency monitoring at the Observatorio Astrofísico de Javalambre for J-PAS and J-PLUS calibration and scheduling

    NASA Astrophysics Data System (ADS)

    Vázquez Ramió, H.; Díaz-Martín, M. C.; Varela, J.; Ederoclite, A.; Maícas, N. Lamadrid, J. L.; Abril, J.; Iglesias-Marzoa, R.; Rodríguez, S.; Tilve, V.; Cenarro, A. J.; Antón Bravo, J. L.; Bello Ferrer, R.; Cristóbal-Hornillos, D.; Guillén Civera, L.; Hernández-Fuertes, J.; Jiménez Mejías, D.; Lasso-Cabrera, N. M.; López Alegre, G.; López Sainz, A.; Luis-Simoes, R. M.; Marín-Franch, A.; Moles, M.; Rueda-Teruel, F.; Rueda-Teruel, S.; Suárez López, O.; Yanes-Díaz, A.

    2015-05-01

    The Javalambre-Physics of the Accelerating Universe Astrophysical Survey (J-PAS; see Benítez et al. 2014) and the Javalambre-Photometric Local Universe Survey (J-PLUS) will be conducted at the brand-new Observatorio Astrofísico de Javalambre (OAJ) in Teruel, Spain. J-PLUS is planned to start by the first half of 2015 while J-PAS first light is expected to happen along 2015. Besides the two main telescopes (with 2.5 m and 80 cm apertures), several smaller-sized facilities are present at the OAJ devoted to site characterization and supporting measurements to be used to calibrate the J-PAS and J-PLUS photometry and to feed up the OAJ's Sequencer with the integrated seeing and the sky transparency. These instruments are: i) an extinction monitor, an 11 " telescope estimating the atmospheric extinction to finally obtain the OAJ extinction curve, which is the initial step to J-PAS overall photometric calibration procedure; ii) an 8 " telescope implementing the Differential Image Motion Monitor (DIMM) technique to obtain the integrated seeing; and iii) an All-Sky Transmission MONitor (ASTMON), a roughly all-sky instrument providing the sky transparency as well as sky brightness and the atmospheric extinction too.

  3. An Investigation of Interrater Reliability for the Rorschach Performance Assessment System (R-PAS) in a Nonpatient U.S. Sample.

    PubMed

    Kivisalu, Trisha M; Lewey, Jennifer H; Shaffer, Thomas W; Canfield, Merle L

    2016-01-01

    The Rorschach Performance Assessment System (R-PAS) aims to provide an evidence-based approach to administration, coding, and interpretation of the Rorschach Inkblot Method (RIM). R-PAS analyzes individualized communications given by respondents to each card to code a wide pool of possible variables. Due to the large number of possible codes that can be assigned to these responses, it is important to consider the concordance rates among different assessors. This study investigated interrater reliability for R-PAS protocols. Data were analyzed from a nonpatient convenience sample of 50 participants who were recruited through networking, local marketing, and advertising efforts from January 2013 through October 2014. Blind recoding was used and discrepancies between the initial and blind coders' ratings were analyzed for each variable with SPSS yielding percent agreement and intraclass correlation values. Data for Location, Space, Contents, Synthesis, Vague, Pairs, Form Quality, Populars, Determinants, and Cognitive and Thematic codes are presented. Rates of agreement for 1,168 responses were higher for more simplistic coding (e.g., Location), whereas agreement was lower for more complex codes (e.g., Cognitive and Thematic codes). Overall, concordance rates achieved good to excellent agreement. Results suggest R-PAS is an effective method with high interrater reliability supporting its empirical basis. PMID:26730817

  4. Crystal structures of the F and pSLT plasmid TraJ N-terminal regions reveal similar homodimeric PAS folds with functional interchangeability

    SciTech Connect

    Lu, Jun; Wu, Ruiying; Adkins, Joshua N.; Joachimiak, Andrzej; Glover, Mark

    2014-09-16

    In the F-family of conjugative plasmids, TraJ is an essential transcriptional activator of the tra operon that encodes most of the proteins required for conjugation. Here we report for the first time the X-ray crystal structures of the TraJ N-terminal regions from the prototypic F plasmid (TraJF11-130) and from the Salmonella virulence plasmid pSLT (TraJpSLT 1-128). Both proteins form similar homodimeric Per-ARNT-Sim (PAS) fold structures. Mutational analysis reveals that the observed dimeric interface is critical for TraJF transcriptional activation, indicating that dimerization of TraJ is required for its in vivo function. An artificial ligand (oxidized dithiothreitol) occupies a cavity in the TraJF dimer interface, while a smaller cavity in corresponding region of the TraJpSLT structure lacks a ligand. Gas chromatography/mass spectrometry-electron ionization analysis of dithiothreitol-free TraJF suggests indole may be the natural TraJ ligand; however, disruption of the indole biosynthetic pathway does not affect TraJF function. Heterologous PAS domains from pSLT and R100 TraJ can functionally replace the TraJF PAS domain, suggesting that TraJ allelic specificity is mediated by the region C-terminal to the PAS domain.

  5. Psychometric Evaluation of a Dutch Version of the Mini PAS-ADD for Assessing Psychiatric Disorders in Adults with Different Levels of Intellectual Disability

    ERIC Educational Resources Information Center

    Janssen, R.; Maes, B.

    2013-01-01

    Background: People with intellectual disabilities (ID) have an increased vulnerability to develop psychiatric problems. Moreover, the early recognition and the accurate diagnosis of psychiatric disorders in the population of persons with ID are challenging. Method: A Dutch version of the Mini PAS-ADD, which is a screening instrument for…

  6. Identification of a novel basic helix-loop-helix-PAS factor, NXF, reveals a Sim2 competitive, positive regulatory role in dendritic-cytoskeleton modulator drebrin gene expression.

    PubMed

    Ooe, Norihisa; Saito, Koichi; Mikami, Nobuyoshi; Nakatuka, Iwao; Kaneko, Hideo

    2004-01-01

    Sim2, a basic helix-loop-helix (bHLH)-PAS transcriptional repressor, is thought to be involved in some symptoms of Down's syndrome. In the course of searching for hypothetical Sim2 relatives, we isolated another bHLH-PAS factor, NXF. NXF was a novel gene and was selectively expressed in neuronal tissues. While no striking homolog of NXF was found in vertebrates, a Caenorhabditis elegans putative transcription factor, C15C8.2, showed similarity in the bHLH-PAS domain. NXF had an activation domain as a transcription activator, and Arnt-type bHLH-PAS subfamily members were identified as the heterodimer partners of NXF. The NXF/Arnt heterodimer was capable of binding and activating a subset of Sim2/Arnt target DNA variants, and Sim2 could compete with the NXF activity on the elements. We showed that Drebrin had several such NXF/Arnt binding elements on the promoter, which could be direct or indirect cross talking points between NXF (activation) and Sim2 (repression) action. Drebrin has been reported to be engaged in dendritic-cytoskeleton modulation at synapses, and such a novel NXF signaling system on neural gene promoter may be a molecular target of the adverse effects of Sim2 in the mental retardation of Down's syndrome. PMID:14701734

  7. One Small Step for the Gram Stain, One Giant Leap for Clinical Microbiology.

    PubMed

    Thomson, Richard B

    2016-06-01

    The Gram stain is one of the most commonly performed tests in the clinical microbiology laboratory, yet it is poorly controlled and lacks standardization. It was once the best rapid test in microbiology, but it is no longer trusted by many clinicians. The publication by Samuel et al. (J. Clin. Microbiol. 54:1442-1447, 2016, http://dx.doi.org/10.1128/JCM.03066-15) is a start for those who want to evaluate and improve Gram stain performance. In an age of emerging rapid molecular results, is the Gram stain still relevant? How should clinical microbiologists respond to the call to reduce Gram stain error rates? PMID:27008876

  8. Histochemical stains as promising means for the laser histochemical surgery of a number of pathologies

    NASA Astrophysics Data System (ADS)

    Piruzyan, L. A.; Mikhailovskiy, Ye. M.; Piruzyan, A. L.

    1999-12-01

    The directions of laboratory and clinical studies oriented to experimental confirmation of the priority concept of `laser histochemical surgery' are presented. The goal of the studies is reproduction on experimental model of a number of pathologies (in vivo and in vitro) of the `sensitization to laser radiation by staining' effect. Testing of the histochemical stains as sensitizers to laser irradiation of their `address substrates', i.e. vitally stained intracellular structures which participate in the pathologic processes evolution is under planning. The processes include: (a) metabolic disorders in the brain cells, i.e. disseminated sclerosis; (b) generalized metabolic disorders- -mucopolysaccharidosis and collagenosises (periarteritis nodosa, rheumatism, rheumatoid arthritis, sclerodermia); (3) metabolic disorders in individual organs--vessel atherosclerosis, hypercholesterolemia, myocardial infarction, cardiosclerosis, caries and parodontosis. The conditions of the studies are detailed in the recommendations along the positions: (1) disease name; (2) disease characteristics: (a) pathomorphologic, (b) biochemical; (3) stains revealing the disease signs and recommended for testing; (4) `address substrates' of the stains that are targets for laser radiation; (5) lasers recommended for the testing after the cells staining in vivo in the corresponding pathology; (6) experimental models of the pathologies suggested for the testing; (7) criteria of the stain efficiency as target sensitizer to the laser light (criteria of the `laser sensitization by staining' efficiency). Possible perspectives for the experimental clinical medicine are indicated of common histochemical stains and lasers use and of practice introduction of the `laser histochemical surgery' in the case the described concept is confirmed in experiments and clinically.

  9. Multispectral image enhancement for H&E stained pathological tissue specimens

    NASA Astrophysics Data System (ADS)

    Bautista, Pinky A.; Abe, Tokiya; Yamaguchi, Masahiro; Ohyama, Nagaaki; Yagi, Yukako

    2008-03-01

    The presence of a liver disease such as cirrhosis can be determined by examining the proliferation of collagen fiber from a tissue slide stained with special stain such as the Masson's trichrome(MT) stain. Collagen fiber and smooth muscle, which are both stained the same in an H&E stained slide, are stained blue and pink respectively in an MT-stained slide. In this paper we show that with multispectral imaging the difference between collagen fiber and smooth muscle can be visualized even from an H&E stained image. In the method M KL bases are derived using the spectral data of those H&E stained tissue components which can be easily differentiated from each other, i.e. nucleus, cytoplasm, red blood cells, etc. and based on the spectral residual error of fiber weighting factors are determined to enhance spectral features at certain wavelengths. Results of our experiment demonstrate the capability of multispectral imaging and its advantage compared to the conventional RGB imaging systems to delineate tissue structures with subtle colorimetric difference.

  10. Acetylcholinesterase staining differentiates functionally distinct auditory pathways in the barn owl.

    PubMed

    Adolphs, R

    1993-03-15

    The aim of this study was to examine how the functional specialization of the barn owl's auditory brainstem might correlate with histochemical compartmentalization. The barn owl uses interaural intensity and time differences to encode, respectively, the vertical and azimuthal positions of sound sources in space. These two auditory cues are processed in parallel ascending pathways that separate from each other at the level of the cochlear nuclei. Sections through the auditory brainstem were stained for acetylcholinesterase (AChE) to examine whether nuclei that process different auditory cues stain differentially for this enzyme. Of the two cochlear nuclei, angularis showed more intense staining than nucleus magnocellularis. Nucleus angularis projects to all of the nuclei and subdivisions of nuclei that belong to the intensity processing pathway. Acetylcholinesterase stained all regions that contain terminal fields of nucleus angularis and thus provided discrimination between the time and intensity pathways. Moreover, staining patterns with acetylcholinesterase were complementary to those previously reported with an anti-calbindin antibody, which stains terminal fields of nucleus laminaris, and thus stains all the nuclei and subdivisions of nuclei that belong to the time pathway. Some of the gross staining patterns observed with AChE were similar to those reported with antibodies to glutamate decarboxylase. However, AChE is a more convenient and definitive marker in discriminating between these pathways than is calbindin or glutamate decarboxylase. Acetylcholinesterase staining of the intensity pathway in the owl may be related to encoding of sound intensity by spike rate over large dynamic ranges. PMID:7681456

  11. Refinements of and commentary on the silver staining techniques of Fernández-Galiano.

    PubMed

    Aufderheide, K J

    2016-07-01

    The original ammoniacal silver carbonate staining technique and subsequent modification developed by Fernández-Galiano are useful for investigating ciliate protozoan systematics and/or ciliate cortical structure and morphogenesis. The technique is complicated, however, by both uncertainties arising from the need to count drops of reagents and subjective control of the staining intensity. I have resolved these complications by defining volumes of reagents rather than using drops and by defining a range of staining times. I also comment on various steps of the techniques. My techniques are simplified and refined to produce consistent, high quality staining results. PMID:27124374

  12. Inadvertent Trypan Blue Staining of Posterior Capsule during Cataract Surgery Associated with "Argentinian Flag" Event.

    PubMed

    Prinzi, Robert A; Alapati, Neeti M; Gappy, Shawn S; Dilly, Jason S

    2016-01-01

    Trypan blue is common in visualizing the anterior capsule during cataract surgery. Inadvertent staining of the posterior capsule during phacoemulsification is a rare complication and there are few reports in the literature. The proposed mechanism of posterior capsule staining in previous reports includes a compromised zonular apparatus or iris retractors facilitating the posterior flow of trypan blue. We report the first case of trypan blue staining of the posterior capsule associated with the "Argentinian flag" sign. In our case, the "Argentinian flag" allowed the trypan blue to seep between the posterior capsule and the lens, staining the anterior surface of the posterior capsule. PMID:27022495

  13. Contrast staining on CT after DSA in ischemic stroke patients progresses to infarction and rarely hemorrhages.

    PubMed

    Amans, Matthew R; Cooke, Daniel L; Vella, Maya; Dowd, Christopher F; Halbach, Van V; Higashida, Randall T; Hetts, Steven W

    2014-01-01

    Contrast staining of brain parenchyma identified on non-contrast CT performed after DSA in patients with acute ischemic stroke (AIS) is an incompletely understood imaging finding. We hypothesize contrast staining to be an indicator of brain injury and suspect the fate of involved parenchyma to be cerebral infarction. Seventeen years of AIS data were retrospectively analyzed for contrast staining. Charts were reviewed and outcomes of the stained parenchyma were identified on subsequent CT and MRI. Thirty-six of 67 patients meeting inclusion criteria (53.7%) had contrast staining on CT obtained within 72 hours after DSA. Brain parenchyma with contrast staining in patients with AIS most often evolved into cerebral infarction (81%). Hemorrhagic transformation was less likely in cases with staining compared with hemorrhagic transformation in the cohort that did not have contrast staining of the parenchyma on post DSA CT (6% versus 25%, respectively, OR 0.17, 95% CI 0.017 - 0.98, p = 0.02). Brain parenchyma with contrast staining on CT after DSA in AIS patients was likely to infarct and unlikely to hemorrhage. PMID:24556308

  14. Contrast Staining on CT after DSA in Ischemic Stroke Patients Progresses to Infarction and Rarely Hemorrhages

    PubMed Central

    Amans, Matthew R.; Cooke, Daniel L.; Vella, Maya; Dowd, Christopher F.; Halbach, Van V.; Higashida, Randall T.; Hetts, Steven W.

    2014-01-01

    Summary Contrast staining of brain parenchyma identified on non-contrast CT performed after DSA in patients with acute ischemic stroke (AIS) is an incompletely understood imaging finding. We hypothesize contrast staining to be an indicator of brain injury and suspect the fate of involved parenchyma to be cerebral infarction. Seventeen years of AIS data were retrospectively analyzed for contrast staining. Charts were reviewed and outcomes of the stained parenchyma were identified on subsequent CT and MRI. Thirty-six of 67 patients meeting inclusion criteria (53.7%) had contrast staining on CT obtained within 72 hours after DSA. Brain parenchyma with contrast staining in patients with AIS most often evolved into cerebral infarction (81%). Hemorrhagic transformation was less likely in cases with staining compared with hemorrhagic transformation in the cohort that did not have contrast staining of the parenchyma on post DSA CT (6% versus 25%, respectively, OR 0.17, 95% CI 0.017 – 0.98, p = 0.02). Brain parenchyma with contrast staining on CT after DSA in AIS patients was likely to infarct and unlikely to hemorrhage. PMID:24556308

  15. Gallocyanin chromalum as a nuclear stain in cytology. I. A cytophotometric comparison of the Husain-Watts Gallocyanin chromalum staining protocol with the Feulgen procedure.

    PubMed

    Schulte, E K; Lyon, H; Prento, P

    1991-05-01

    In the present study, the staining characteristics of the Gallocyanin chromalum technique devised by Husain and Watts are compared with the Feulgen reaction. Liver imprints, blood smears, and cervical smears were fixed in ethanol and stained with either the Husain and Watts Gallocyanin chromalum reagent or the Feulgen-Schiff reagent. The slides were then post-treated with 70% ethanol-HCl pH 1.0, or with phosphotungstic acid for 0.5-30 min. The integrated optical density of cell nuclei was measured with a VIDAS image analyzer. In the material stained with the Husain and Watts procedure, some Gallocyanin chromalum was removed from the nuclei in the early phase (5 min) of all the post-treatment steps, followed by a plateau phase where the integrated optical density remained constant for 30 min. In this phase, the nuclear absorbance was highly reproducible and of the same size regardless of the post-treatment. Both the Husain and Watts procedure and the Feulgen-reaction gave quantitative staining of DNA. The Gallocyanin chromalum stain after Husain and Watts is a quick staining procedure for quantitative evaluation of DNA in cytological material. Proper rinsing of the slides is necessary for a good reproducibility of results. PMID:1723725

  16. Morphological Alterations of the Palpebral Conjunctival Epithelium in a Dry Eye Model

    PubMed Central

    Henriksson, Johanna Tukler; De Paiva, Cintia S.; Farley, William; Pflugfelder, Stephen C.; Burns, Alan R.; Bergmanson, Jan P.G.

    2012-01-01

    Purpose To investigate the normal palpebral conjunctival histology in C57BL/6 mice, and the structural changes that occur in a dry eye model. Methods 24 male and female C57BL/6 mice, 8 untreated (UT) and 16 exposed to experimental ocular surface desiccating stress (DS). Ocular dryness was induced by administration of scopolamine hydrobromide (0.5 mg/0.2 ml) QID for 5 (DS5) or 10 (DS10) days. Counts and measurements were obtained using anatomical reference points and goblet cell density was investigated with a variety of stains. Results Near the junction between the lid margin and the normal palpebral conjunctiva, the epithelium had an average thickness of 45.6±10.5μm, 8.8±2.0 cell layers, versus 37.7±5.6μm, 7.4±1.3 layers in DS10 (P<0.05). In the goblet cell populated palpebral region the normal epithelium was thicker (P<0.05) than in DS5 and DS10. In the control, 43% of the goblet cells were covered by squamous epithelium, compared to 58% (DS5) and 63% (DS10) (P<0.05). A decreased number of Periodic Acid Schiff (PAS) and Alcian blue stained goblet cells was observed in the dry eye. Not all goblet cells stained with PAS and Alcian blue. Conclusions The mouse palpebral conjunctival epithelium was structurally similar to the human. After DS the palpebral conjunctival epithelium decreased in thickness and goblet cell access to the surface appeared to be inhibited by surrounding epithelial cells, potentially slowing down their migration to the surface. Differential staining with PAS and Alcian blue suggests there may be different subtypes of conjunctival goblet cells. PMID:23146932

  17. Differences in Memory Development among C57BL/6NCrl, 129S2/SvPasCrl, and FVB/NCrl Mice after Delay and Trace Fear Conditioning

    PubMed Central

    March, Amelia; Borchelt, David; Golde, Todd; Janus, Christopher

    2014-01-01

    Fear-conditioning testing paradigms have been used to study differences in memory formation between inbred mouse strains, including numerous mouse models of human diseases. In this study, we characterized the conditioned fear memory of 3 inbred strains: C57BL/6NCrl, 129S2/SvPasCrl, and FVB/NCrl, obtained from Charles River Laboratories. We used 2 training paradigms: delay conditioning, in which an unconditional stimulus coterminates with the presentation of a conditional stimulus, and trace conditioning, in which the conditional and unconditional stimuli are separated by a trace interval. In each paradigm, we evaluated the recent (3 d) and remote (25 d) memory of the mice by using a longitudinal design. Our results showed that both C57BL/6NCrl and 129S2/SvPasCrl mice developed strong and long-lasting context and tone memories in both paradigms, but FVB/NCrl mice showed a weaker but nevertheless consistent tone memory after delay training. Tone memory in the FVB strain was stronger in male than female mice. The remote tone memory of 129S2/SvPasCrl mice diminished after delay training but was stable and stronger than that of C57BL/6NCrl mice after trace training. In conclusion, both C57BL/6NCrl and 129S2/SvPasCrl mice showed reliable and long-lasting fear memory after delay or trace training, with 129 mice showing particularly strong tone memory after trace conditioning. The FVB/NCrl strain, especially male mice, showed reliable tone fear memory after delay training. Our findings confirm that both C57BL/6NCrl and 129S2/SvPasCrl mice develop strong context and tone memory in delay and trace fear-conditioning paradigms. PMID:24672832

  18. Differences in memory development among C57BL/6NCrl, 129S2/SvPasCrl, and FVB/NCrl mice after delay and trace fear conditioning.

    PubMed

    March, Amelia; Borchelt, David; Golde, Todd; Janus, Christopher

    2014-02-01

    Fear-conditioning testing paradigms have been used to study differences in memory formation between inbred mouse strains, including numerous mouse models of human diseases. In this study, we characterized the conditioned fear memory of 3 inbred strains: C57BL/6NCrl, 129S2/SvPasCrl, and FVB/NCrl, obtained from Charles River Laboratories. We used 2 training paradigms: delay conditioning, in which an unconditional stimulus coterminates with the presentation of a conditional stimulus, and trace conditioning, in which the conditional and unconditional stimuli are separated by a trace interval. In each paradigm, we evaluated the recent (3 d) and remote (25 d) memory of the mice by using a longitudinal design. Our results showed that both C57BL/6NCrl and 129S2/SvPasCrl mice developed strong and long-lasting context and tone memories in both paradigms, but FVB/NCrl mice showed a weaker but nevertheless consistent tone memory after delay training. Tone memory in the FVB strain was stronger in male than female mice. The remote tone memory of 129S2/SvPasCrl mice diminished after delay training but was stable and stronger than that of C57BL/6NCrl mice after trace training. In conclusion, both C57BL/6NCrl and 129S2/SvPasCrl mice showed reliable and long-lasting fear memory after delay or trace training, with 129 mice showing particularly strong tone memory after trace conditioning. The FVB/NCrl strain, especially male mice, showed reliable tone fear memory after delay training. Our findings confirm that both C57BL/6NCrl and 129S2/SvPasCrl mice develop strong context and tone memory in delay and trace fear-conditioning paradigms. PMID:24672832

  19. Glycoprotein secretion in a tracheal organ culture system

    SciTech Connect

    Warunek, D.J.

    1985-01-01

    Glycoprotein secretion in the rat trachea was studied in vitro, utilizing a modified, matrix embed/perfusion chamber. Baseline parameters of the culture environment were determined by enzymatic and biochemical procedures. The effect of pilocarpine on the release of labelled glycoproteins from the tracheal epithelium was assessed. After a single stimulation with the drug, there was a significant increase in the release of /sup 14/C-glucosamine and /sup 3/H-fucose-labelled glycoprotein. The response was dose-dependent. Similar results were obtained after a second exposure to pilocarpine. However, no dose response was observed. Morphological analyses of the tracheal epithelial secretory cells by Alcian Blue/Periodic Acid Schiff staining showed a significant decrease in the total number of Alcian Blue staining cells and an increase in the mixed cell population after a single exposure to pilocarpine. Second stimulation with the drug showed that the trachea was able to respond again, this time with a further decrease in the number of Alcian Blue staining cells and a decrease in the PAS staining cells as well. Carbohydrate analyses after the first simulation with pilocarpine showed increased levels of N-acetyl neuraminic acid and the neutral carbohydrates, fucose and galactose, in the precipitated glycoproteins.

  20. Evaluation of Intestinal Protozoan Morphology in Human Fecal Specimens Preserved in EcoFix: Comparison of Wheatley’s Trichrome Stain and EcoStain

    PubMed Central

    Garcia, Lynne S.; Shimizu, Robyn Y.

    1998-01-01

    As a result of disposal problems related to the use of mercury compounds, many laboratories have switched from mercuric chloride-based Schaudinn’s and polyvinyl alcohol (PVA) stool preservatives to other, non-mercury-based preservatives. A comparison of organism recoveries and morphologies of the intestinal protozoa was undertaken with PVA containing the EcoFix zinc-based Schaudinn’s preservative (Meridian Diagnostics, Inc.); both Wheatley’s modification of Gomori’s trichrome stain (WT) and EcoStain (ES) were used to stain 51 human fecal specimens. Morphology, clarity of nuclear and cytoplasmic detail, overall color differences, and the ease or difficulty in detecting intestinal protozoa in fecal debris were assessed for the two permanent stained smears. Overall, organism morphology of the intestinal protozoa stained with WT and that of protozoa stained with ES were not equal in nuclear and cytoplasmic detail or range of color. However, the same organisms were identified in stained fecal smears with either WT or ES, with the exception of situations in which organism numbers were characterized as rare. Included were 67 protozoan challenges (number of organisms): Entamoeba histolytica-Entamoeba dispar (5), Entamoeba coli (9), Entamoeba hartmanni (6), Endolimax nana (12), Iodamoeba bütschlii (8), Blastocystis hominis (19), Giardia lamblia (6), Dientamoeba fragilis (2), yeast (2), and leukocytes (2). Five specimens were negative for parasites but contained fecal debris that was compared for morphologic detail and color range. The ES produces a more gray-green monotone with very little pink or red tone; contrast among the various colors is less than that seen with WT. Stain intensity for all organisms was acceptable, and there were no problems with stain deposition. The quality of the protozoan morphology with ES was often comparable to that with WT (36 of 67 [53.7%]) and, in some cases, better (24 of 67 [35.8%]). Organisms on the WT-stained smear exhibited

  1. Evaluation of intestinal protozoan morphology in human fecal specimens preserved in EcoFix: comparison of Wheatley's trichrome stain and EcoStain.

    PubMed

    Garcia, L S; Shimizu, R Y

    1998-07-01

    As a result of disposal problems related to the use of mercury compounds, many laboratories have switched from mercuric chloride-based Schaudinn's and polyvinyl alcohol (PVA) stool preservatives to other, non-mercury-based preservatives. A comparison of organism recoveries and morphologies of the intestinal protozoa was undertaken with PVA containing the EcoFix zinc-based Schaudinn's preservative (Meridian Diagnostics, Inc.); both Wheatley's modification of Gomori's trichrome stain (WT) and EcoStain (ES) were used to stain 51 human fecal specimens. Morphology, clarity of nuclear and cytoplasmic detail, overall color differences, and the ease or difficulty in detecting intestinal protozoa in fecal debris were assessed for the two permanent stained smears. Overall, organism morphology of the intestinal protozoa stained with WT and that of protozoa stained with ES were not equal in nuclear and cytoplasmic detail or range of color. However, the same organisms were identified in stained fecal smears with either WT or ES, with the exception of situations in which organism numbers were characterized as rare. Included were 67 protozoan challenges (number of organisms): Entamoeba histolytica-Entamoeba dispar (5), Entamoeba coli (9), Entamoeba hartmanni (6), Endolimax nana (12), Iodamoeba bütschlii (8), Blastocystis hominis (19), Giardia lamblia (6), Dientamoeba fragilis (2), yeast (2), and leukocytes (2). Five specimens were negative for parasites but contained fecal debris that was compared for morphologic detail and color range. The ES produces a more gray-green monotone with very little pink or red tone; contrast among the various colors is less than that seen with WT. Stain intensity for all organisms was acceptable, and there were no problems with stain deposition. The quality of the protozoan morphology with ES was often comparable to that with WT (36 of 67 [53.7%]) and, in some cases, better (24 of 67 [35.8%]). Organisms on the WT-stained smear exhibited better

  2. Staining of proteins in gels with Coomassie G-250 without organic solvent and acetic acid.

    PubMed

    Lawrence, Ann-Marie; Besir, H Uuml Seyin

    2009-01-01

    In classical protein staining protocols using Coomassie Brilliant Blue (CBB), solutions with high contents of toxic and flammable organic solvents (Methanol, Ethanol or 2-Propanol) and acetic acid are used for fixation, staining and destaining of proteins in a gel after SDS-PAGE. To speed up the procedure, heating the staining solution in the microwave oven for a short time is frequently used. This usually results in evaporation of toxic or hazardous Methanol, Ethanol or 2-Propanol and a strong smell of acetic acid in the lab which should be avoided due to safety considerations. In a protocol originally published in two patent applications by E.M. Wondrak (US2001046709 (A1), US6319720 (B1)), an alternative composition of the staining solution is described in which no organic solvent or acid is used. The CBB is dissolved in bidistilled water (60-80 mg of CBB G-250 per liter) and 35 mM HCl is added as the only other compound in the staining solution. The CBB staining of the gel is done after SDS-PAGE and thorough washing of the gel in bidistilled water. By heating the gel during the washing and staining steps, the process can be finished faster and no toxic or hazardous compounds are evaporating. The staining of proteins occurs already within 1 minute after heating the gel in staining solution and is fully developed after 15-30 min with a slightly blue background that is destained completely by prolonged washing of the stained gel in bidistilled water, without affecting the stained protein bands. PMID:19684570

  3. Cascaded solar photovoltaic cells based on InP-GaInPAs and InP-CdS heterostructures

    SciTech Connect

    Allakhverdiyev, A.M.; Andreyev, V.; Guseynov, I.A.; Iventyeva, O.O.; Ismailov, V.I.

    1984-08-01

    Cascaded solar cells with different widths of the forbidden band narrow band and wide band were studied. The narrow band cell was based on nInP-n(p)GaInPAs-pInP structure, and the wide band cell was based on pInP-nCdS heterostructure with similar lattice parameters of the contacting materials. The narrow band cells were made by liquid phase epitaxy, and the wide band cells were made by gas transport precipitation of CdS. The spectral characteristics show the feasibility of expanding the region of photosensitivity by using the principle of cascaded conversion of solar energy. The cell with pn junction in Ga0.5In0.75As0.85P0.42 utilizes light with photon energies of less than 1.35 eV that have passed without absorption through the covering photocell. The longwave limits of photosensitivity are determined by the width of the forbidden band of the active region of the corresponding photocells. The shortwave end of the spectrum is stretched out as a result of the low surface recombination rate.

  4. Fault activity in the epicentral area of the 1580 Dover Strait (Pas-de-Calais) earthquake (northwestern Europe)

    NASA Astrophysics Data System (ADS)

    García-Moreno, D.; Verbeeck, K.; Camelbeeck, T.; De Batist, M.; Oggioni, F.; Zurita Hurtado, O.; Versteeg, W.; Jomard, H.; Collier, J. S.; Gupta, S.; Trentesaux, A.; Vanneste, K.

    2015-05-01

    On 1580 April 6 one of the most destructive earthquakes of northwestern Europe took place in the Dover Strait (Pas de Calais). The epicentre of this seismic event, the magnitude of which is estimated to have been about 6.0, has been located in the offshore continuation of the North Artois shear zone, a major Variscan tectonic structure that traverses the Dover Strait. The location of this and two other moderate magnitude historical earthquakes in the Dover Strait suggests that the North Artois shear zone or some of its fault segments may be presently active. In order to investigate the possible fault activity in the epicentral area of the AD 1580 earthquake, we have gathered a large set of bathymetric and seismic-reflection data covering the almost-entire width of the Dover Strait. These data have revealed a broad structural zone comprising several subparallel WNW-ESE trending faults and folds, some of them significantly offsetting the Cretaceous bedrock. The geophysical investigation has also shown some indication of possible Quaternary fault activity. However, this activity only appears to have affected the lowermost layers of the sediment infilling Middle Pleistocene palaeobasins. This indicates that, if these faults have been active since Middle Pleistocene, their slip rates must have been very low. Hence, the AD 1580 earthquake appears to be a very infrequent event in the Dover Strait, representing a good example of the moderate magnitude earthquakes that sometimes occur in plate interiors on faults with unknown historical seismicity.

  5. The necessity of sample quality assessment in 14C AMS dating: The case of Cova des Pas (Menorca - Spain)

    NASA Astrophysics Data System (ADS)

    Van Strydonck, Mark; Boudin, Mathieu; Guerrero Ayuso, Víctor M.; Calvo, Manuel; Fullola, Josep M.; Àngeles Petit, M.

    2010-04-01

    The Cova des Pas cave is a karstic cave in the cliffs of the Barranc (canyon) de Trebalúger. It is a small cave only 6.5 m deep, 4.5 m wide and 1.7 m high. Yet more than 70 burials, in foetal position, from the Early Iron Age were found in this small cave. The conservation of the archaeological remains was very unusual. Not only wood, ropes and other plant material was found, but also remains of body tissue, hair and leather. In spite of the remarkable preservation of the bodies and artefacts, the state of conservation of the bone material was very bad. The bones contained little and heavily deteriorated collagen and the organic plant material was very fragile. The special environmental conditions of the cave are the cause of these unusual preservation conditions. Although the cave is situated in a limestone cliff, the soil of the cave is very acid. Unexpectedly bad sample quality seemed to be the major drawback for AMS radiocarbon dating on hair, wood, ropes and bone collagen, as well as on bio-apatite.

  6. PAS7 encodes a novel yeast member of the WD-40 protein family essential for import of 3-oxoacyl-CoA thiolase, a PTS2-containing protein, into peroxisomes.

    PubMed Central

    Marzioch, M; Erdmann, R; Veenhuis, M; Kunau, W H

    1994-01-01

    To identify components of the peroxisomal import pathway in yeast, we have isolated pas mutants affected in peroxisome biogenesis. Two mutants assigned to complementation group 7 define a new gene, PAS7, whose product is necessary for import of thiolase, a PTS2-containing protein, but not for that of SKL (PTS1)-containing proteins, into peroxisomes. We have cloned PAS7 by complementation of the oleic acid non-utilizing phenotype of the pas7-1 strain. The DNA sequence predicts a 42.3 kDa polypeptide of 375 amino acids encoding a novel member of the beta-transducin related (WD-40) protein family. A Myc epitope-tagged Pas7p, expressed under the control of the CUP1 promotor, was functionally active. Subcellular localization studies revealed that in the presence of thiolase this epitope-tagged Pas7p in part associates with peroxisomes. However, in a thiolase-deficient mutant, Pas7p was entirely found in the cytoplasm. We suggest that Pas7p mediates the binding of thiolase to these organelles. Images PMID:7957058

  7. Kinetic silver staining and quantification of proteins adsorbed to microtiter plates.

    PubMed

    Root, D D; Wang, K

    1993-03-01

    A silver stain was used to detect and quantitate proteins adsorbed to microtiter plate wells. The kinetics of the development of the silver stain were analyzed with an automated microtiter plate reader. The lag time for stain development was found to be a consistent indicator of the amount of protein adsorbed to a microtiter plate well. Protein which was not preadsorbed to the microtiter plate was not effectively stained by silver. Complete adsorption of protein applied to the microtiter plate was possible by drying small amounts of protein in very dilute buffers. Variations in sensitivity for different proteins were less than 30% for the panel of proteins examined. Determinations from kinetic silver staining agreed with those from copper staining for bovine albumin adsorbed to microtiter plates. The precision of kinetic silver staining assay was optimal in the range of 40 to 200 ng per microtiter plate well. In this range, the standard deviations averaged less than 5%. Even smaller amounts of protein can be detected and interpolated down to approximately 10 ng per well. The kinetic silver staining method can be used on standard microtiter plate readers without special filters and is readily adaptable to automated systems. PMID:8470810

  8. Analysis of Staining Observed on Structures in the Georgetown, South Carolina Area

    SciTech Connect

    Cramer, Stephen D.; Covino, Bernard S. Jr.; Govier, R. Dale

    2002-05-01

    Beginning around 1970, the Georgetown, SC, community complained about black dust and red stains collecting on houses, cars, boats, and other structures. The community, through the South Carolina Department of Health and Environmental Control (SCDHEC), seeks to identify the source or cause of the staining and ways to reduce or eliminate it in the future.

  9. Histopathological Findings in Immunohistological Staining of the Granulomatous Tissue Reaction Associated with Tuberculosis

    PubMed Central

    Karimi, Shirin; Pourabdollah, Mihan; Sadr, Makan; Karbasi, Mehrdad; Bahadori, Moslem

    2014-01-01

    Purpose. The histological diagnosis of Mycobacterium tuberculosis (MTB) remains a diagnostic challenge despite different methods. Immunohistochemistry (IHC) not only could confirm granulomatous tissue involvement but also can demonstrate MTB antigen immunolocalization. This study tries to clarify the details of immunohistochemical staining for MTB with pAbBCG. Materials/Methods. Twenty-three confirmed TB granulomatous tissue samples were studied by Ziehl-Neelsen and immunohistochemistry (IHC) staining with pAbBCG. Samples were selected from the archive of the Department of Pathology, National Research Institute of Tuberculosis and Lung Disease, Tehran, Iran. Results. IHC staining was positive in all samples, whereas Ziehl-Neelsen was positive in 9 cases out of 23 (39.1%). Tissue types used were pleural tissue, lymph nodes, and lung tissue. IHC showed positive coarse granular cytoplasmic and round, fragmented bacillary staining. In this study, epithelioid cells clearly showed more positive staining at the periphery of the granuloma rather than the center of granuloma. There is also positive staining in endothelial cells, fibroblasts, plasma cells, lymphocytes, and macrophages outside the granuloma. Conclusion. Considering the criteria of positive immunohistochemical staining of TB granulomatous reactions, this stain not only highlights the presence of mycobacterial antigens for tissue diagnosis, but also could morphologically localize its distribution in different cells. PMID:24511393

  10. Ultrastructural study of Chlamydia trachomatis surface antigens by immunogold staining with monoclonal antibodies.

    PubMed Central

    Kuo, C C; Chi, E Y

    1987-01-01

    Surface antigens of Chlamydia trachomatis were studied by immunogold staining with monoclonal antibodies and by electron microscopy. The serovar- and subspecies-specific epitopes were the most surface accessible. The species- and genus-specific epitopes were the least surface exposed. Similar serological specificity as that in the microimmunofluorescence test was demonstrated by immunogold staining. Images PMID:2437035

  11. Human Hemochromatosis Protein (HFE) Immunoperoxidase Stain Highlights Choriocarcinoma within Mixed Germ Cell Tumors.

    PubMed

    Cox, Jesse L; Talmon, Geoffrey A; Koepsell, Scott A

    2016-01-01

    Identification of choriocarcinoma within a germ cell tumor can have major implications for the subsequent staging and treatment of testicular neoplasms. Immunoperoxidase staining greatly enhances the speed and sensitivity of identifying occult, though clinically significant, tumor components. In mixed germ cell tumors, staining for beta-human chorionic gonadotropin (β-hCG) has been historically used to assess for the presence and burden of choriocarcinoma. However, current β-hCG stains produce variable, intense staining of trophoblastic elements and surrounding tissues, clouding the assessment of true-positive staining. Human hemochromatosis protein (HFE) is a membrane bound mediator of iron transport expressed at high levels within placenta. Additionally, previous reports have demonstrated that choriocarcinoma cell lines express HFE, although in vivo expression had not been examined. To address whether HFE can stain trophoblastic elements, HFE immunohistochemistry was conducted in choriocarcinoma (n = 4), mixed germ cell tumors (n = 11), seminoma (n = 4), and placenta (n = 11). HFE consistently demonstrated cytoplasmic and membranous staining, highlighting both syncytiotrophoblasts and cytotrophoblasts within choriocarcinoma and placenta. Staining of intratumoral white blood cells was observed within seminomas and mixed germ cell tumors, corroborating prior reports stating that HFE highlights monocytes and macrophages. Taken together, HFE may serve as an alternative target from β-hCG for immunoperoxidase studies when highlighting choriocarcinoma. PMID:27034532

  12. Human Hemochromatosis Protein (HFE) Immunoperoxidase Stain Highlights Choriocarcinoma within Mixed Germ Cell Tumors

    PubMed Central

    Talmon, Geoffrey A.; Koepsell, Scott A.

    2016-01-01

    Identification of choriocarcinoma within a germ cell tumor can have major implications for the subsequent staging and treatment of testicular neoplasms. Immunoperoxidase staining greatly enhances the speed and sensitivity of identifying occult, though clinically significant, tumor components. In mixed germ cell tumors, staining for beta-human chorionic gonadotropin (β-hCG) has been historically used to assess for the presence and burden of choriocarcinoma. However, current β-hCG stains produce variable, intense staining of trophoblastic elements and surrounding tissues, clouding the assessment of true-positive staining. Human hemochromatosis protein (HFE) is a membrane bound mediator of iron transport expressed at high levels within placenta. Additionally, previous reports have demonstrated that choriocarcinoma cell lines express HFE, although in vivo expression had not been examined. To address whether HFE can stain trophoblastic elements, HFE immunohistochemistry was conducted in choriocarcinoma (n = 4), mixed germ cell tumors (n = 11), seminoma (n = 4), and placenta (n = 11). HFE consistently demonstrated cytoplasmic and membranous staining, highlighting both syncytiotrophoblasts and cytotrophoblasts within choriocarcinoma and placenta. Staining of intratumoral white blood cells was observed within seminomas and mixed germ cell tumors, corroborating prior reports stating that HFE highlights monocytes and macrophages. Taken together, HFE may serve as an alternative target from β-hCG for immunoperoxidase studies when highlighting choriocarcinoma. PMID:27034532

  13. Tear staining in pigs: a potential tool for welfare assessment on commercial farms

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tear staining or chromodacryorrhea refers to a dark stain below the inner corner of the eye, caused by porphyrin-pigmented secretion from the Harderian gland. It been shown to be a consistent indicator of stress in rats, and recently it has been shown to correlate with social stress and a barren env...

  14. A Modified Verhoeff-Van Gieson Elastin Histochemical Stain to Enable Pulmonary Arterial Hypertension Model Characterization

    PubMed Central

    Percival, K.R.; Radi, Z.A.

    2016-01-01

    Optimal histochemical staining is critical to ensure excellent quality stained sections to enable light microscopic and histomorphometric image analysis. Verhoeff-van Gieson is the most widely used histochemical stain for the visualization of vascular elastic fibers. However, it is notoriously difficult to differentiate fine elastic fibers of small vasculature to enable histomorphometric image analysis, especially in organs such as the lung. A tissue fixation procedure of 10% neutral buffered formalin with subsequent fixation in 70% ethanol further compounds the problem of small vessel staining and identification. Therefore, a modified Verhoeff’s elastin stain was developed as a reliable method to optimally highlight the internal and external elastic laminae of small arteries (50-100 µm external diameter) and intra-acinar vessels (10-50 µm external diameter) in 3 µm thick lung tissue sections from models of pulmonary arterial hypertension. This modified Verhoeff’s elastin stain demonstrated well-defined staining of fine elastic fibers of pulmonary blood vessels enabling subsequent histomorphometric image analysis of vessel wall thickness in small arteries and intra-acinar vessels. In conclusion, modification of the standard Verhoeff-van Gieson histochemical stain is needed to visualize small caliber vessels’ elastic fibers especially in tissues fixed in 10% neutral buffered formalin followed by additional fixation in 70% ethanol. PMID:26972717

  15. A simplified method for differential staining of aborted and non-aborted pollen grains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ability to use chemical staining to discriminate aborted from non-aborted pollen grains has well-known practical applications in agriculture. A commonly used technique for assessing pollen vitality, Alexander’s stain, uses chloral hydrate, phenol and mercuric chloride, all of which are highly to...

  16. Human herpesvirus 8 and iron staining are useful in differentiating Kaposi sarcoma from interstitial granuloma annulare.

    PubMed

    Wada, David A; Perkins, Sherrie L; Tripp, Sheryl; Coffin, Cheryl M; Florell, Scott R

    2007-02-01

    We studied 20 granuloma annulare (GA) cases (10 interstitial and 10 palisaded) and 19 Kaposi sarcoma (KS) cases (9 "early" and 10 typical). Tissue sections were stained for iron, Hale colloidal iron, human herpesvirus 8 (HHV-8), CD31, CD34, CD68, collagen IV, factor XIIIa, and MIB-1. Iron staining of dermal tissue associated with the lesion was confirmed in all KS cases and no GA cases. Immunohistochemical stains for HHV-8 were positive in all 9 cases of early KS and most cases (9/10) of typical KS. All 20 cases of GA were HHV-8-. CD31, CD34, CD68, factor XIIIa, and MIB-1 were also stained but were difficult to interpret and did not seem specific for GA or KS. Iron staining and immunohistochemical HHV-8 staining in combination were reliable markers for KS compared with interstitial GA. MIB-1 fractions of less than 5% favored a diagnosis of GA, whereas fractions greater than 10% favored a diagnosis of KS. This study provides novel data characterizing iron staining in KS and details the use of iron staining, HHV-8, and MIB-1 to distinguish KS from GA. PMID:17210517

  17. 2D Source area prediction based on physical characteristics of a regular, passive blood drip stain.

    PubMed

    Basu, Nabanita; Bandyopadhyay, Samir Kumar

    2016-09-01

    Violent criminal acts are often accompanied by dynamic blood shedding events. Bloodstain pattern analysis particularly deals with estimation of the dynamic blood shedding events from the static bloodstain patterns that have been left at the scene. Of all the stain patterns present at a crime scene, drip stain patterns are common stain patterns one would expect to document at a violent crime scene. The paper documents statistically significant correlations between different physical parameters, such as fall height, total number of spines associated with each stain. Statistical significant correlation between the angle of impact and the total number of spines associated with each stain pattern has been established in this work. The paper propounds that the breadth of a regular drip stain is particularly significant in making predictions empirically as also statistically about the surface area from which blood has dripped leading to the formation of a particular drip stain. A data model has been developed using machine learning techniques to predict the range of surface radius from which blood has dripped and lead to the formation of a particular drip stain (Accuracy: 97.53%, Sensitivity=0.9481, Specificity=1). PMID:27295073

  18. Picrosirius red staining: a useful tool to appraise collagen networks in normal and pathological tissues.

    PubMed

    Lattouf, Raed; Younes, Ronald; Lutomski, Didier; Naaman, Nada; Godeau, Gaston; Senni, Karim; Changotade, Sylvie

    2014-10-01

    Specific staining of the extracellular matrix components is especially helpful in studying tissue remodeling, particularly in the case of connective tissue pathologies. As developed by Junqueira and colleagues in 1979, specific staining by Picrosirius red is one of the most important stains to study collagen networks in different tissues. Under polarized light, collagen bundles appear green, red or yellow, and are easily differentiated from the black background, thus allowing for quantitative morphometric analysis. As Junqueira and colleagues point out, many studies use color staining to differentiate collagen bundles and to specify collagen types, yet other studies report that polarized colors only reflect fiber thickness and packing. Using a simple histological example, our study illustrates the inability of Picrosirius red staining to differentiate collagen types, since the absorbed amount of polarized light by this dye strictly depends on the orientation of the collagen bundles. PMID:25023614

  19. A Versatile Method for Immunofluorescent Staining of Cells Cultured on Permeable Membrane Inserts.

    PubMed

    Gillespie, Jenni L; Anyah, Anwuli; Taylor, John M; Marlin, Jerry W; Taylor, Tracey A H

    2016-01-01

    BACKGROUND Obtaining high-quality images of cellular structures via immunofluorescence staining is critical for cellular localization studies. Often, these studies cannot be performed in parallel with certain oncology, virology, pharmacokinetic, and drug absorption studies due to model system technicalities requiring the cells to be cultured on porous membranes rather than glass or plastic. MATERIAL AND METHODS Here, we report a method of immunofluorescent staining of cells cultured on permeable membranes. RESULTS As proof of principle, HeLa cells grown on Transwell® membrane supports were stained with fluorescently labeled antibodies using this modified immunofluorescence staining method and visualized by fluorescent microscopy. CONCLUSIONS This protocol is a convenient alternative to staining cells on glass coverslips, thereby expanding the scope and applications of this important research tool. PMID:27616137

  20. An automatic stain removal algorithm of series aerial photograph based on flat-field correction

    NASA Astrophysics Data System (ADS)

    Wang, Gang; Yan, Dongmei; Yang, Yang

    2010-10-01

    The dust on the camera's lens will leave dark stains on the image. Calibrating and compensating the intensity of the stained pixels play an important role in the airborne image processing. This article introduces an automatic compensation algorithm for the dark stains. It's based on the theory of flat-field correction. We produced a whiteboard reference image by aggregating hundreds of images recorded in one flight and use their average pixel values to simulate the uniform white light irradiation. Then we constructed a look-up table function based on this whiteboard image to calibrate the stained image. The experiment result shows that the proposed procedure can remove lens stains effectively and automatically.

  1. Advanced negative detection method comparable to silver stain for SDS-PAGE separated proteins detection.

    PubMed

    Wang, Xu; Hwang, Sun-Young; Cong, Wei-Tao; Jin, Li-Tai; Choi, Jung-Kap

    2016-10-01

    In order to achieve an easy, rapid and sensitive protocol to detect proteins in polyacrylamide gel, an advanced negative detection method comparable to silver stain is described. When a gel was incubated with Phloxine B and followed by the development in acidic solution, the zones where forming protein-dye complex were selectively transparent, unlike opaque gel background. Within 50 min after electrophoresis, down to 0.1-0.4 ng of gel-separated proteins (similar with silver stain) could be observed, without labor-intensive and time-consuming procedure. Comparing with the most common negative stain method, Imidazole-zinc stain, Phloxine B stain has been shown higher sensitivity and distinct contrast between the transparent protein bands/spots and opaque background than those; furthermore, it is no longer necessary to concern about retention time of observation. This technique may provide a sensitive and practical choice for proteomics researches. PMID:27430933

  2. Tooth stain during 2 years' use of chlorhexidine- and fluoride-containing dentifrices.

    PubMed

    Dolles, O K; Eriksen, H M; Gjermo, P

    1979-08-01

    The purpose of the present study was to evaluate the effect of fluoride upon the stain-inducing capacity of chlorhexidine. Ninety-one children, 13 years of age, volunteered for the experiment. Three test dentifrices were used, containing 0.1% NaF, 0.1% NaF + 2% chlorhexidine, and 2% chlorhexidine respectively. The experiment was carried out as a double-blind test, and the degree of staining was recorded after 1 and 2 years. Four methods were used for stain scoring. The results after 1 year showed that chlorhexidine induced less stain when applied together with fluoride than when applied alone. After 2 years this difference disappeared, indicating that fluoride only retarded chlorhexidine-induced staining of teeth. PMID:295486

  3. Porcine Intestinal Mast Cells. Evaluation of Different Fixatives for Histochemical Staining Techniques Considering Tissue Shrinkage

    PubMed Central

    Rieger, J.; Twardziok, S.; Huenigen, H.; Hirschberg, R.M.; Plendl, J.

    2013-01-01

    Staining of mast cells (MCs), including porcine ones, is critically dependent upon the fixation and staining technique. In the pig, mucosal and submucosal MCs do not stain or stain only faintly after formalin fixation. Some fixation methods are particularly recommended for MC staining, for example the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF) has been reported to work excellently for the preservation of fixation-sensitive antigens. The aim of this study was to establish a reliable histological method for counting of MCs in the porcine intestinum. For this purpose, different tissue fixation and staining methods that also allow potential subsequent immunohistochemical investigations were evaluated in the porcine mucosa, as well as submucosa of small and large intestine. Tissues were fixed in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with either polychromatic methylene blue, alcian blue or toluidine blue. For the first time our study reveals that ZSF, a heavy metal fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not suitable for histochemical visualization of MCs in the pig intestine. All other tested fixatives were suitable. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs difficult. The polychromatic methylene blue proved to be the optimal staining. In order to compare MC counting results of the different fixation methods, tissue shrinkage was taken into account. As even the same fixation caused shrinkagedifferences between tissue from small and large intestine, different factors for each single fixation and intestinal localization had to be calculated. Tissue shrinkage varied between 19% and 57%, the highest tissue shrinkage was found after fixation with ZSF in the large intestine, the lowest one in the small intestine after lead acetate fixation. Our study emphasizes that MC counting results from data using different

  4. Porcine intestinal mast cells. Evaluation of different fixatives for histochemical staining techniques considering tissue shrinkage.

    PubMed

    Rieger, J; Twardziok, S; Huenigen, H; Hirschberg, R M; Plendl, J

    2013-01-01

    Staining of mast cells (MCs), including porcine ones, is critically dependent upon the fixation and staining technique. In the pig, mucosal and submucosal MCs do not stain or stain only faintly after formalin fixation. Some fixation methods are particularly recommended for MC staining, for example the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF) has been reported to work excellently for the preservation of fixation-sensitive antigens. The aim of this study was to establish a reliable histological method for counting of MCs in the porcine intestinum. For this purpose, different tissue fixation and staining methods that also allow potential subsequent immunohistochemical investigations were evaluated in the porcine mucosa, as well as submucosa of small and large intestine. Tissues were fixed in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with either polychromatic methylene blue, alcian blue or toluidine blue. For the first time our study reveals that ZSF, a heavy metal fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not suitable for histochemical visualization of MCs in the pig intestine. All other tested fixatives were suitable. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs difficult. The polychromatic methylene blue proved to be the optimal staining. In order to compare MC counting results of the different fixation methods, tissue shrinkage was taken into account. As even the same fixation caused shrinkage-differences between tissue from small and large intestine, different factors for each single fixation and intestinal localization had to be calculated. Tissue shrinkage varied between 19% and 57%, the highest tissue shrinkage was found after fixation with ZSF in the large intestine, the lowest one in the small intestine after lead acetate fixation. Our study emphasizes that MC counting results from data using

  5. [Suicide risk and suicide attempt in North Pas de Calais Region. Lessons from the survey Mental Health in General Population].

    PubMed

    Danel, T; Vilain, J; Roelandt, J L; Salleron, J; Vaiva, G; Amariei, A; Amarie, A; Plancke, L; Plance, L; Duhamel, A

    2010-01-01

    The Santé Mentale en Population Générale Survey (Mental Health in General Population Survey (MHGP)) is a multicentre international research and action project initiated by the World Health Organisation Collaboration Centre for research and training in mental health. Its aims are to assess the prevalence of the major mental health disorders in the general adult population and from this to record perceptions associated with "mental illness", "madness" and "depression" together with different means of assistance and specialist or lay care. In this work we present the analysis of data on risks of suicide and past history of suicide attempts in the Nord pas de Calais region. We present the qualitative features of these phenomena and correlations with socio-economic, cultural and psychopathological factors, which are discussed in terms of both protective and vulnerability factors. Risk of suicide is present in 15% of the Nord pas de Calais population and is divided into 10.44% slight risk, 2.37% moderate risk and 2.2% high risk. A comparison with data from the MHGP survey in other regions reveals the high risk of suicide in the NPDC region. A risk of suicide is present is 13% of the population in other SMPG survey regions, broken down into 9.1% low risk, 2.1% medium risk and 1.7% high risk. Compared to the 2.2% high risk figure for NPDC, the population in this category is 21% larger. In terms of risk and protective factors, a bivariate analysis of socio-economic and cultural factors confirms the classical risk factors of sex, marital, occupational and educational status and income. The odds-ratio for these socio-economic and cultural factors can be calculated from logistic regression and the protective factors ranked in decreasing order from religion (Muslim versus other religions), martial status (marked versus separated), age (over 58 years old), occupational status (working or retired versus unemployed), income (more than 1300 euros versus less than 840 euros), sex

  6. Automatic disease screening method using image processing for dried blood microfluidic drop stain pattern recognition.

    PubMed

    Sikarwar, Basant S; Roy, Mukesh; Ranjan, Priya; Goyal, Ayush

    2016-07-01

    This paper examines programmed automatic recognition of infection from samples of dried stains of micro-scale drops of patient blood. This technique has the upside of being low-cost and less-intrusive and not requiring puncturing the patient with a needle for drawing blood, which is especially critical for infants and the matured. It also does not require expensive pathological blood test laboratory equipment. The method is shown in this work to be successful for ailment identification in patients suffering from tuberculosis and anaemia. Illness affects the physical properties of blood, which thus influence the samples of dried micro-scale blood drop stains. For instance, if a patient has a severe drop in platelet count, which is often the case of dengue or malaria patients, the blood's physical property of viscosity drops substantially, i.e. the blood is thinner. Thus, the blood micro-scale drop stain samples can be utilised for diagnosing maladies. This paper presents programmed automatic examination of the dried micro-scale drop blood stain designs utilising an algorithm based on pattern recognition. The samples of micro-scale blood drop stains of ordinary non-infected people are clearly recognisable as well as the samples of micro-scale blood drop stains of sick people, due to key distinguishing features. As a contextual analysis, the micro-scale blood drop stains of patients infected with tuberculosis have been contrasted with the micro-scale blood drop stains of typical normal healthy people. The paper dives into the fundamental flow mechanics behind how the samples of the dried micro-scale blood drop stain is shaped. What has been found is a thick ring like feature in the dried micro-scale blood drop stains of non-ailing people and thin shape like lines in the dried micro-scale blood drop stains of patients with anaemia or tuberculosis disease. The ring like feature at the periphery is caused by an outward stream conveying suspended particles to the edge

  7. Hardware and software facilities for the J-PAS and J-PLUS surveys archiving, processing and data publication

    NASA Astrophysics Data System (ADS)

    Cristóbal-Hornillos, D.; Varela, J.; Ederoclite, A.; Vázquez Ramió, H.; López-Sainz, A.; Hernández-Fuertes, J.; Civera, T.; Muniesa, D.; Moles, M.; Cenarro, A. J.; Marín-Franch, A.; Yanes-Díaz, A.

    2015-05-01

    The Observatorio Astrofísico de Javalambre consists of two main telescopes: JST/T250, a 2.5 m telescope with a FoV of 3 deg, and JAST/T80, a 83 cm with a 2 deg FoV. JST/T250 will be devoted to complete the Javalambre-PAU Astronomical Survey (J-PAS). It is a photometric survey with a system of 54 narrow-band plus 3 broad-band filters covering an area of 8500°^2. The JAST/T80 will perform the J-PLUS survey, covering the same area in a system of 12 filters. This contribution presents the software and hardware architecture designed to store and process the data. The processing pipeline runs daily and it is devoted to correct instrumental signature on the science images, to perform astrometric and photometric calibration, and the computation of individual image catalogs. In a second stage, the pipeline performs the combination of the tile mosaics and the computation of final catalogs. The catalogs are ingested in as Scientific database to be provided to the community. The processing software is connected with a management database to store persistent information about the pipeline operations done on each frame. The processing pipeline is executed in a computing cluster under a batch queuing system. Regarding the storage system, it will combine disk and tape technologies. The disk storage system will have capacity to store the data that is accessed by the pipeline. The tape library will store and archive the raw data and earlier data releases with lower access frequency.

  8. Genetic Evidence for Function of the bHLH-PAS Protein Gce/Met As a Juvenile Hormone Receptor.

    PubMed

    Jindra, Marek; Uhlirova, Mirka; Charles, Jean-Philippe; Smykal, Vlastimil; Hill, Ronald J

    2015-07-01

    Juvenile hormones (JHs) play a major role in controlling development and reproduction in insects and other arthropods. Synthetic JH-mimicking compounds such as methoprene are employed as potent insecticides against significant agricultural, household and disease vector pests. However, a receptor mediating effects of JH and its insecticidal mimics has long been the subject of controversy. The bHLH-PAS protein Methoprene-tolerant (Met), along with its Drosophila melanogaster paralog germ cell-expressed (Gce), has emerged as a prime JH receptor candidate, but critical evidence that this protein must bind JH to fulfill its role in normal insect development has been missing. Here, we show that Gce binds a native D. melanogaster JH, its precursor methyl farnesoate, and some synthetic JH mimics. Conditional on this ligand binding, Gce mediates JH-dependent gene expression and the hormone's vital role during development of the fly. Any one of three different single amino acid mutations in the ligand-binding pocket that prevent binding of JH to the protein block these functions. Only transgenic Gce capable of binding JH can restore sensitivity to JH mimics in D. melanogaster Met-null mutants and rescue viability in flies lacking both Gce and Met that would otherwise die at pupation. Similarly, the absence of Gce and Met can be compensated by expression of wild-type but not mutated transgenic D. melanogaster Met protein. This genetic evidence definitively establishes Gce/Met in a JH receptor role, thus resolving a long-standing question in arthropod biology. PMID:26161662

  9. An adaptive algorithm for detection of multiple-type, positively stained nuclei in IHC images with minimal prior information: application to OLIG2 staining gliomas

    NASA Astrophysics Data System (ADS)

    Akakin, Hatice C.; Gokozan, Hamza; Otero, Jose; Gurcan, Metin N.

    2015-03-01

    We propose a method to detect and segment the oligodendrocytes and gliomas in OLIG2 immunoperoxidase stained tissue sections. Segmentation of cell nuclei is essential for automatic, fast, accurate and consistent analysis of pathology images. In general, glioma cells and oligodendrocytes mostly differ in shape and size within the tissue slide. In OLIG2 stained tissue images, gliomas are represented with irregularly shaped nuclei with varying sizes and brown shades. On the other hand, oligodendrocytes have more regular round nuclei shapes and are smaller in size when compared to glioma cells found in oligodendroglioma, astrocytomas, or oligoastrocytomas. The first task is to detect the OLIG2 positive cell regions within a region of interest image selected from a whole slide. The second task is to segment each cell nucleus and count the number of cell nuclei. However, the cell nuclei belonging to glioma cases have particularly irregular nuclei shapes and form cell clusters by touching or overlapping with each other. In addition to this clustered structure, the shading of the brown stain and the texture of the nuclei differ slightly within a tissue image. The final step of the algorithm is to classify glioma cells versus oligodendrocytes. Our method starts with color segmentation to detect positively stained cells followed by the classification of single individual cells and cell clusters by K-means clustering. Detected cell clusters are segmented with the H-minima based watershed algorithm. The novel aspects of our work are: 1) the detection and segmentation of multiple-type, positively-stained nuclei by incorporating only minimal prior information; and 2) adaptively determining clustering parameters to adjust to the natural variation in staining as well as the underlying cellular structure while accommodating multiple cell types in the image. Performance of the algorithm to detect individual cells is evaluated by sensitivity and precision metrics. Promising

  10. Acridine orange staining reaction as an index of physiological activity in Escherichia coli

    NASA Technical Reports Server (NTRS)

    McFeters, G. A.; Singh, A.; Byun, S.; Callis, P. R.; Williams, S.

    1991-01-01

    The assumption that the acridine orange (AO) color reaction may be used as an index of physiological activity was investigated in laboratory grown Escherichia coli. Spectrofluorometric observations of purified nucleic acids, ribosomes and the microscopic color of bacteriophage-infected cells stained with AO confirmed the theory that single-stranded nucleic acids emit orange to red fluorescence while those that are double-stranded fluoresce green in vivo. Bacteria growing actively in a rich medium could be distinguished from cells in stationary phase by the AO reaction. Cells from log phase appeared red, whereas those in stationary phase were green. However, this differentiation was not seen when the bacteria were grown in a minimal medium or when a variation of the staining method was used. Also, shifting bacteria in stationary phase to starvation conditions rapidly changed their AO staining reaction. Boiling and exposure to lethal concentrations of azide and formalin resulted in stationary-phase cells that appeared red after staining but bacteria killed with chlorine remained green. These findings indicate that the AO staining reaction may be suggestive of physiological activity under defined conditions. However, variables in staining and fixation procedures as well as uncertainties associated with mixed bacterial populations in environmental samples may produce results that are not consistent with the classical interpretation of this reaction. The importance of validating the putative physiological implications of this staining reaction is stressed.

  11. Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition

    PubMed Central

    Booth, David S.; Avila-Sakar, Agustin; Cheng, Yifan

    2011-01-01

    Single particle electron microscopy (EM), of both negative stained or frozen hydrated biological samples, has become a versatile tool in structural biology 1. In recent years, this method has achieved great success in studying structures of proteins and macromolecular complexes 2, 3. Compared with electron cryomicroscopy (cryoEM), in which frozen hydrated protein samples are embedded in a thin layer of vitreous ice 4, negative staining is a simpler sample preparation method in which protein samples are embedded in a thin layer of dried heavy metal salt to increase specimen contrast 5. The enhanced contrast of negative stain EM allows examination of relatively small biological samples. In addition to determining three-dimensional (3D) structure of purified proteins or protein complexes 6, this method can be used for much broader purposes. For example, negative stain EM can be easily used to visualize purified protein samples, obtaining information such as homogeneity/heterogeneity of the sample, formation of protein complexes or large assemblies, or simply to evaluate the quality of a protein preparation. In this video article, we present a complete protocol for using an EM to observe negatively stained protein sample, from preparing carbon coated grids for negative stain EM to acquiring images of negatively stained sample in an electron microscope operated at 120kV accelerating voltage. These protocols have been used in our laboratory routinely and can be easily followed by novice users. PMID:22215030

  12. The application of visible wavelength reflectance hyperspectral imaging for the detection and identification of blood stains.

    PubMed

    Li, Bo; Beveridge, Peter; O'Hare, William T; Islam, Meez

    2014-12-01

    Current methods of detection and identification of blood stains rely largely on visual examination followed by presumptive tests such as Kastle-Meyer, Leuco-malachite green or luminol. Although these tests are useful, they can produce false positives and can also have a negative impact on subsequent DNA tests. A novel application of visible wavelength reflectance hyperspectral imaging has been used for the detection and positive identification of blood stains in a non contact and non destructive manner on a range of coloured substrates. The identification of blood staining was based on the unique visible absorption spectrum of haemoglobin between 400 and 500 nm. Images illustrating successful discrimination of blood stains from nine red substances are included. It has also been possible to distinguish between blood and approximately 40 other reddish stains. The technique was also successfully used to detect latent blood stains deposited on white filter paper at dilutions of up to 1 in 512 folds and on red tissue at dilutions of up to 1 in 32 folds. Finally, in a blind trial, the method successfully detected and identified a total of 9 blood stains on a red T-shirt. PMID:25498930

  13. Subjective and objective measures of corneal staining related to multipurpose care systems.

    PubMed

    Pritchard, Nicola; Young, Graeme; Coleman, Sarah; Hunt, Chris

    2003-03-01

    An objective, digital-imaging method of measuring corneal staining was evaluated in 24 subjects wearing soft contact lenses. The method was used to compare the clinical performance of common multipurpose care systems (MPS) for soft contact lens care. Subjects used three different MPS, one containing polyquaternium-1 (PQ) and two containing polyhexanide (PX1 and PX2), for 2 weeks in a randomised, single-masked (investigator) crossover study. Corneal staining induced with the three MPS was analysed using an image-processing program (ImageTool, UTHSCSA Version 2, University of Texas, USA). Conjunctival hyperaemia and papillae were also evaluated. The intraclass correlation coefficient was similar with image analysis to that of investigator grading (0.876, 0.879, respectively). Significant differences in staining response were detected using the objective method. There was significantly less staining area with polyquaternium-1 (PQ) than polyhexanide (PQ: 0.12 mm(2), PX2: 0.91 mm(2)). Inferior palpebral papillae were significantly greater with PX2 than with PQ (1.0, 0.7 (0-4), respectively). The technique was shown to be an effective method of evaluating different corneal staining responses. Bilateral corneal staining in three or more quadrants is useful in the diagnosis of MPS-related staining. PMID:16303491

  14. Peripheral Insertion of a Central Venous Access Device Under Fluoroscopic Guidance Using a Peripherally Accessed System (PAS) Port in the Forearm

    SciTech Connect

    Hata, Yasuhiro; Morita, Sojiro; Morita, Yoshitaka; Awatani, Toshihide; Takasaki, Motohiro; Horimi, Tadashi; Ozawa, Zen

    1998-05-15

    Purpose: We describe the technique, efficacy, and complications of fluoroscopy-guided implantation of a central venous access device using a peripherally accessed system (PAS) port via the forearm. Methods: Beginning in July 1994, 105 central venous access devices were implanted in 104 patients for the long-term infusion of antibiotics or antineoplasmic agents, blood products, or parenteral nutrition. The devices was inserted under fluoroscopic guidance with real-time venography from a peripheral route. Results: All ports were successfully implanted. There were no procedure-related complications. No thrombosis or local infection was observed; however, in six patients catheter-related phlebitis occurred. Conclusion: Fluoroscopy-guided implantation of a central venous access device using a PAS port via the forearm is safe and efficacious, and injection of contrast medium through a peripheral IV catheter before introduction of the catheter helps to avoid catheter-related phlebitis.

  15. Characterization and developmental expression of xSim, a Xenopus bHLH/PAS gene related to the Drosophila neurogenic master gene single-minded.

    PubMed

    Coumailleau, P; Penrad-Mobayed, M; Lecomte, C; Bollérot, K; Simon, F; Poellinger, L; Angelier, N

    2000-12-01

    We have isolated a novel gene from Xenopus, denominated xSim, which encodes a protein of 760 amino acids containing a basic helix-loop-helix (bHLH) motif contiguous to a PAS domain characteristic of an emerging family of transcriptional regulators so called bHLH/PAS. xSim shares a strong amino acid sequence identity with the Drosophila Single-minded (dSim) and with the murine Sim1 and Sim2 proteins. Phylogenetic analysis reveals that xSim gene is an ortholog gene of the mSim2 gene. Spatio-temporal analysis shows a maternal and a zygotic expression of xSim throughout early Xenopus development. In situ hybridization assays reveal that the transcripts are enriched in the animal hemisphere until blastula stage and extend to the marginal zone at early gastrula stage. As development proceeds, xSim is mainly restricted to the central nervous system. PMID:11091086

  16. Tear Film, Contact Lens, and Patient Factors Associated with Corneal Staining

    PubMed Central

    Sinnott, Loraine T.

    2011-01-01

    Purpose. The purpose of this study was to examine ocular surface and tear film, contact lens, care solution, medical, and patient-related factors that are associated with corneal staining in contact lens wearers. Methods. In this cross-sectional/nested case–control study, in addition to the assessment of corneal staining with fluorescein, a variety of tear film and ocular surface, contact lens, and patient-related factors were examined. Poisson regression models were used to examine the relation between corneal staining and these factors. Results. Data from 413 patients were eligible for the analyses described. The average age was 30.6 ± 11.1 years, and 277 (67.1%) of the patients were women. Several factors were shown to be related to increased corneal staining in multivariate modeling, including increased daily wearing times (P = 0.0006), lower income (P = 0.0008), lissamine green conjunctival staining (P = 0.002), contact lens deposition (P = 0.007), increased tear meniscus height (P = 0.007), and decreased hydrogel nominal water content (P = 0.02). The wearing of silicone hydrogels (as opposed to hydrogels) was protective against corneal staining (P = 0.0004). Notably, neither contact lens care solutions nor disinfectants were associated with corneal staining. Conclusions. Corneal staining in contact lens wearers continues to be a frequent, but not well understood, outcome. These data suggest that contact lens factors (water content, material, wearing time, and deposition) are more generally associated with corneal staining than are contact lens care solutions or other ocular surface and tear film, demographic, or medical factors. PMID:21087960

  17. Effect of Staining Solutions on Color Stability of Silorane & Methacrylate Restorative Material

    PubMed Central

    S. Madhyastha, Prashanthi; G. Naik, Dilip; Kotian, Ravindra; Srikant, N.; M. R. Bhat, Kumar

    2015-01-01

    Color stability throughout the functional lifetime of restorations is important for the durability of treatment and of cosmetic importance. The purpose of this study was to evaluate the discoloration properties of a silorane-based (Filtek P90) and methacrylate-based (Z100) composites upon exposure to different staining solutions that are used on day to day basis (turmeric, tea, coffee, cocoa, lime, yoghurt and distilled water) for different immersion periods (1, 7, 14 and 28 days). The colors of all specimens before and after storage in the solutions were measured by a reflectance spectrophotometer based on CIE Lab system and the color differences were calculated. Data were statistically analyzed by repeated measures of ANOVA and sidak post hoc test (for immersion period);‘t’ test (for each material) and one way ANOVA (for staining agents). All the staining agents showed significant difference in staining over time in both the materials. However, Z100 showed higher quantum of discoloration at all time periods at each staining agents (p<0.005). In conclusion, the silorane-based resin (Filtek P90) composites exhibited better color stability (less change in ΔE) after exposure to the staining solutions. Among the staining agents cocoa was found to be least staining followed by lime, yoghurt, coffee, tea whereas turmeric discolored the composites to the maximum. Highest discoloration was seen at day 28 in all staining agents. Cocoa and lime discolored to maximum at early stages but remained stable thereafter whereas tea, coffee and turmeric progressively discolored the composite over time.

  18. Brain damage in a large cohort of solvent abusers.

    PubMed

    Al-Hajri, Zahra; Del Bigio, Marc R

    2010-04-01

    The neuropathology of solvent inhalation consists of patchy myelin loss with white matter macrophages that contain granular inclusions. It has been described only in a small number of cases. We sought to characterize the abnormalities in greater detail. In a retrospective study from 1995 to 2009, we encountered 88 autopsy cases with documented history of solvent abuse by inhalation and 1 with industrial exposure. Among these are 6 fetuses and infants with maternal exposure, 23 children (12-17 years), and 60 adults (18-66 years). Available brain samples from 75 cases were stained with solochrome cyanein (to demonstrate myelin) and periodic acid-Schiff (PAS) (to highlight the inclusions). Forty brains of ethanol and/or illicit drug exposed individuals and ten cases of multiple sclerosis were examined as controls. We found that 16 cases (age 23-49, median 37 years) had well-established leukoencephalopathy with multifocal myelin loss and abundant macrophages that stain with PAS and which contain birefringent inclusions. Six cases (age 15-55, median 27 years) had early leukoencephalopathy with scattered macrophages but no obvious myelin changes. Clusters of PAS-staining but non-birefringent macrophages were seen in 2/10 cases of (active) multiple sclerosis and in none of the ethanol/drug exposed brains. Ultrastructurally, inclusions from solvent cases differed from multiple sclerosis cases. Although exposure to solvents is impossible to quantify, there appears to be a duration-dependent effect. Brain damage related to solvent abuse can begin within only a few years of the onset. In the context of substance abuse, the changes are relatively specific for solvent inhalation and do not appear to result from demyelination alone. Interaction with ethanol cannot be excluded as a compounding risk factor. PMID:20300918

  19. Mechanisms of airway responses to esophageal acidification in cats.

    PubMed

    Lang, Ivan M; Haworth, Steven T; Medda, Bidyut K; Forster, Hubert; Shaker, Reza

    2016-04-01

    Acid in the esophagus causes airway constriction, tracheobronchial mucous secretion, and a decrease in tracheal mucociliary transport rate. This study was designed to investigate the neuropharmacological mechanisms controlling these responses. In chloralose-anesthetized cats (n = 72), we investigated the effects of vagotomy or atropine (100 μg·kg(-1)·30 min(-1) iv) on airway responses to esophageal infusion of 0.1 M PBS or 0.1 N HCl at 1 ml/min. We quantified 1) diameter of the bronchi, 2) tracheobronchial mucociliary transport rate, 3) tracheobronchial mucous secretion, and 4) mucous content of the tracheal epithelium and submucosa. We found that vagotomy or atropine blocked the airway constriction response but only atropine blocked the increase in mucous output and decrease in mucociliary transport rate caused by esophageal acidification. The mucous cells of the mucosa produced more Alcian blue- than periodic acid-Schiff (PAS)-stained mucosubstances, and the mucous cells of the submucosa produced more PAS- than Alcian blue-stained mucosubstances. Selective perfusion of the different segments of esophagus with HCl or PBS resulted in significantly greater production of PAS-stained mucus in the submucosa of the trachea adjacent to the HCl-perfused esophagus than in that adjacent to the PBS-perfused esophagus. In conclusion, airway constriction caused by esophageal acidification is mediated by a vagal cholinergic pathway, and the tracheobronchial transport response is mediated by cholinergic receptors. Acid perfusion of the esophagus selectively increases production of neutral mucosubstances of the apocrine glands by a local mechanism. We hypothesize that the airway responses to esophageal acid exposure are part of the innate, rather than acute emergency, airway defense system. PMID:26846551

  20. The preparation of cervical scrape material for automated cytology using gallocyanin chrome-alum stain.

    PubMed

    Eason, P J; Tucker, J H

    1979-01-01

    A method is described for preparing cervical scrape specimens for automated analysis on the Cerviscan prescreening system. In order to reduce the cellular clumping found in cervical scrape material, cells are collected in suspension, syringed to disaggregate the cell clumps, and then pipetted onto a glass to give a monolayer of cells. The cells are then stained with gallocyanin chrome-alum to give the required quantitation of nucleic acid content, using a rapid staining procedure. Experimental results are given which show that specimens prepared by this method are more suitable for automated analysis than the conventional Papanicolaou stained preparation. PMID:86562

  1. Highly sensitive fluorescent stain for detecting lipopolysaccharides in sodium dodecyl sulfate polyacrylamide gel electrophoresis.

    PubMed

    Wang, Xu; Zhou, Ayi; Cai, Wanhui; Yu, Dongdong; Zhu, Zhongxin; Jiang, Chengxi; Jin, Litai

    2015-08-01

    A sensitive and simple technique was developed for the visualization of gel-separated lipopolysaccharides by using a hydrazide derivative, UGF202. As low as 0.5-1 ng total LPS could be detected by UGF202 stain, which is 2- and 16-fold more sensitive than that of the commonly used Pro-Q Emerald 300 and Keenan et al. developed silver stain, respectively. The results indicated that UGF202 stain could be a good choice for LPS determination in polyacrylamide gels. PMID:25930092

  2. Large-volume en-bloc staining for electron microscopy-based connectomics

    PubMed Central

    Hua, Yunfeng; Laserstein, Philip; Helmstaedter, Moritz

    2015-01-01

    Large-scale connectomics requires dense staining of neuronal tissue blocks for electron microscopy (EM). Here we report a large-volume dense en-bloc EM staining protocol that overcomes the staining gradients, which so far substantially limited the reconstructable volumes in three-dimensional (3D) EM. Our protocol provides densely reconstructable tissue blocks from mouse neocortex sized at least 1 mm in diameter. By relaxing the constraints on precise topographic sample targeting, it makes the correlated functional and structural analysis of neuronal circuits realistic. PMID:26235643

  3. SiR–Hoechst is a far-red DNA stain for live-cell nanoscopy

    PubMed Central

    Lukinavičius, Gražvydas; Blaukopf, Claudia; Pershagen, Elias; Schena, Alberto; Reymond, Luc; Derivery, Emmanuel; Gonzalez-Gaitan, Marcos; D'Este, Elisa; Hell, Stefan W.; Wolfram Gerlich, Daniel; Johnsson, Kai

    2015-01-01

    Cell-permeable DNA stains are popular markers in live-cell imaging. Currently used DNA stains for live-cell imaging are either toxic, require illumination with blue light or are not compatible with super-resolution microscopy, thereby limiting their utility. Here we describe a far-red DNA stain, SiR–Hoechst, which displays minimal toxicity, is applicable in different cell types and tissues, and is compatible with super-resolution microscopy. The combination of these properties makes this probe a powerful tool for live-cell imaging. PMID:26423723

  4. Development of a Whole Blood Staining Device for use During Space Shuttle Flights

    NASA Technical Reports Server (NTRS)

    Sams, Clarence F.; Crucian, Brian E.; Clift, Vaughan L.; Meinelt, Ellen M.

    1999-01-01

    Exposure to microgravity during space flight results in profound physiologic changes. Numerous studies have shown changes in circulating populations of peripheral blood immune cells immediately after space flight. It is currently unknown if these changes result from exposure to microgravity or are caused by the stress of reentry and readaptation to gravity. We have developed the whole blood staining device as a system for the staining of whole blood collected during space flight for subsequent flow cytometric analysis, This device contains all liquids to address safety issues concerned with space flight and also moves the cells through the staining, lyse/fixation and dilution steps.

  5. SiR-Hoechst is a far-red DNA stain for live-cell nanoscopy.

    PubMed

    Lukinavičius, Gražvydas; Blaukopf, Claudia; Pershagen, Elias; Schena, Alberto; Reymond, Luc; Derivery, Emmanuel; Gonzalez-Gaitan, Marcos; D'Este, Elisa; Hell, Stefan W; Gerlich, Daniel Wolfram; Johnsson, Kai

    2015-01-01

    Cell-permeable DNA stains are popular markers in live-cell imaging. Currently used DNA stains for live-cell imaging are either toxic, require illumination with blue light or are not compatible with super-resolution microscopy, thereby limiting their utility. Here we describe a far-red DNA stain, SiR-Hoechst, which displays minimal toxicity, is applicable in different cell types and tissues, and is compatible with super-resolution microscopy. The combination of these properties makes this probe a powerful tool for live-cell imaging. PMID:26423723

  6. A concurrent comparison of isoniazid plus PAS with three regimens of isoniazid alone in the domiciliary treatment of pulmonary tuberculosis in South India

    PubMed Central

    1960-01-01

    Recent studies have shown that treatment of pulmonary tuberculosis with isoniazid plus p-aminosalicylic acid (PAS) at home is, in the majority of cases, as satisfactory as treatment with the same combination of drugs in sanatorium and does not appear to expose the patient's contacts to any special risk. Before mass domiciliary chemotherapy can be introduced, however, a question that has to be decided is what drug or drugs and what dosage and rhythm of administration will be most effective. This paper presents the results of a controlled comparison of four chemotherapeutic regimens: (a) 3.9-5.5 mg/kg body-weight of isoniazid plus 0.2-0.3 g/kg body-weight of PAS (sodium salt) daily in two doses (the standard combined chemotherapy); (b) 7.8-9.6 mg/kg body-weight of isoniazid alone daily in one dose; (c) 7.8-9.6 mg/kg body-weight of isoniazid alone daily in two doses; (d) 3.9-5.5 mg/kg body-weight of isoniazid alone daily in two doses. Isoniazid plus PAS proved to be the most satisfactory regimen; it was clinically effective and there were very few toxic manifestations. PMID:14447270

  7. PAS kinase is activated by direct SNF1-dependent phosphorylation and mediates inhibition of TORC1 through the phosphorylation and activation of Pbp1

    PubMed Central

    DeMille, Desiree; Badal, Bryan D.; Evans, J. Brady; Mathis, Andrew D.; Anderson, Joseph F.; Grose, Julianne H.

    2015-01-01

    We describe the interplay between three sensory protein kinases in yeast: AMP-regulated kinase (AMPK, or SNF1 in yeast), PAS kinase 1 (Psk1 in yeast), and the target of rapamycin complex 1 (TORC1). This signaling cascade occurs through the SNF1-dependent phosphorylation and activation of Psk1, which phosphorylates and activates poly(A)- binding protein binding protein 1 (Pbp1), which then inhibits TORC1 through sequestration at stress granules. The SNF1-dependent phosphorylation of Psk1 appears to be direct, in that Snf1 is necessary and sufficient for Psk1 activation by alternate carbon sources, is required for altered Psk1 protein mobility, is able to phosphorylate Psk1 in vitro, and binds Psk1 via its substrate-targeting subunit Gal83. Evidence for the direct phosphorylation and activation of Pbp1 by Psk1 is also provided by in vitro and in vivo kinase assays, including the reduction of Pbp1 localization at distinct cytoplasmic foci and subsequent rescue of TORC1 inhibition in PAS kinase–deficient yeast. In support of this signaling cascade, Snf1-deficient cells display increased TORC1 activity, whereas cells containing hyperactive Snf1 display a PAS kinase–dependent decrease in TORC1 activity. This interplay between yeast SNF1, Psk1, and TORC1 allows for proper glucose allocation during nutrient depletion, reducing cell growth and proliferation when energy is low. PMID:25428989

  8. Tissue staining properties of lectins from the seeds of the jack fruit (Artocarpus integrifolia) and the winged bean (Psophocarpus tetragonolobus).

    PubMed

    Vijayakumar, T; Robertson, D; McIntosh, D; Forrester, J A

    1987-01-01

    N-acetyl-D-galactosamine binding lectins from winged bean (Psophocarpus tetragonolobus) and jack fruit (Artocarpus integrifolia) were isolated, purified and conjugated with horse radish peroxidase and their tissue staining properties studied. Despite having an apparently common inhibiting sugar, the lectins showed differences in their staining properties. The lectin from the winged bean stained none of the mouse and human tissues tried even after neuraminidase treatment whereas the jack fruit lectin stained most of the untreated cells. The staining was found to be improved by the prior treatment of the cells with neuraminidase and inhibited completely by the inhibiting sugar. The differences in the staining properties of the lectins are discussed. PMID:2452865

  9. The Effects of Maekmoondong-Tang on Cockroach Extract-Induced Allergic Asthma

    PubMed Central

    Sohn, Sung-Hwa; Jung, Kyung-Hwa; Lee, Kun-young; Yeom, Yu Rim; Kim, Gae-Eun; Jung, Sungki; Jung, Heejae; Bae, Hyunsu

    2014-01-01

    Maekmoondong-tang (MMDT) has long been used in Asian countries to treat respiratory diseases. However, the precise mechanisms underlying its effects on asthma are unknown. This study was conducted to evaluate the protective effects of MMDT in a cockroach allergen (CKA-)induced animal model of allergic asthma. After being challenged with CKA, the number of macrophages, eosinophils, neutrophils, lymphocytes, and total cells in the bronchoalveolar lavage fluid (BALF) was evaluated. The Th2 specific cytokines IL-4, IL-5, and IL-13 were also analyzed in BALF along with IgE levels in serum. For histological analysis, hematoxylin and eosin (H&E) staining, periodic acid-Schiff (PAS) staining, and immunohistochemical staining were performed. In addition, airway hyperresponsiveness was assessed by noninvasive plethysmography. The cellular profiles and histopathologic analysis demonstrated that peribronchial and perivascular inflammatory cell infiltrates were significantly decreased in the MMDT-treated groups compared with the cockroach extract-injected (CKA) groups. In addition, the IgE, IL-4, IL-5, and IL-13 levels were significantly decreased in the MMDT group. MMDT treatment also significantly attenuated airway hyperresponsiveness. These results demonstrated that MMDT significantly reduced the hallmark signs of asthma: elevated serum IgE, airway eosinophilia, airway remodeling, mucus hypersecretion, and airway hyperresponsiveness. The remarkable antiasthmatic effects of MMDT suggest its therapeutic potential in allergic asthma treatment. PMID:24723965

  10. Systematic investigation of drip stains on apparel fabrics: The effects of prior-laundering, fibre content and fabric structure on final stain appearance.

    PubMed

    de Castro, Therese C; Taylor, Michael C; Kieser, Jules A; Carr, Debra J; Duncan, W

    2015-05-01

    Bloodstain pattern analysis is the investigation of blood deposited at crime scenes and the interpretation of that pattern. The surface that the blood gets deposited onto could distort the appearance of the bloodstain. The interaction of blood and apparel fabrics is in its infancy, but the interaction of liquids and apparel fabrics has been well documented and investigated in the field of textile science (e.g. the processes of wetting and wicking of fluids on fibres, yarns and fabrics). A systematic study on the final appearance of drip stains on torso apparel fabrics (100% cotton plain woven, 100% polyester plain woven, blend of polyester and cotton plain woven and 100% cotton single jersey knit) that had been laundered for six, 26 and 52 cycles prior to testing was investigated in the paper. The relationship between drop velocity (1.66±0.50m/s, 4.07±0.03m/s, 5.34±0.18m/s) and the stain characteristics (parent stain area, axes 1 and 2 and number of satellite stains) for each fabric was examined using analysis of variance. The experimental design and effect of storing blood were investigated on a reference sample, which indicated that the day (up to five days) at which the drops were generated did not affect the bloodstain. The effect of prior-laundering (six, 26 and 52 laundering cycles), fibre content (cotton vs. polyester vs. blend) and fabric structure (plain woven vs. single jersey knit) on the final appearance of the bloodstain were investigated. Distortion in the bloodstains produced on non-laundered fabrics indicated the importance of laundering fabrics to remove finishing treatments before conducting bloodstain experiments. For laundered fabrics, both the cotton fabrics and the blend had a circular to oval stain appearance, while the polyester fabric had a circular appearance with evidence of spread along the warp and weft yarns, which resulted in square-like stains at the lowest drop velocity. A significant (p<0.001) increase in the stain size on

  11. Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction

    SciTech Connect

    Cardoso, Rosa; Love, Robert; Nilsson, Carol L; Bergqvist, Simon; Nowlin, Dawn; Yan, Jiangli; Liu, Kevin K.-C.; Zhu, Jing; Chen, Ping; Deng, Ya-Li; Dyson, H Jane; Greig, Michael J; Brooun, Alexei

    2012-11-09

    The heterodimer HIF-1α (hypoxia inducible factor)/HIF-β (also known as ARNT-aryl hydrocarbon nuclear translocator) is a key mediator of cellular response to hypoxia. The interaction between these monomer units can be modified by the action of small molecules in the binding interface between their C-terminal heterodimerization (PasB) domains. Taking advantage of the presence of several cysteine residues located in the allosteric cavity of HIF-1α PasB domain, we applied a cysteine-based reactomics “hotspot identification” strategy to locate regions of HIF-1α PasB domain critical for its interaction with ARNT. COMPOUND 5 was identified using a mass spectrometry-based primary screening strategy and was shown to react specifically with Cys255 of the HIF-1α PasB domain. Biophysical characterization of the interaction between PasB domains of HIF-1α and ARNT revealed that covalent binding of COMPOUND 5 to Cys255 reduced binding affinity between HIF-1α and ARNT PasB domains approximately 10-fold. Detailed NMR structural analysis of HIF-1α-PasB-COMPOUND 5 conjugate showed significant local conformation changes in the HIF-1α associated with key residues involved in the HIF-1α/ARNT PasB domain interaction as revealed by the crystal structure of the HIF-1α/ARNT PasB heterodimer. Our screening strategy could be applied to other targets to identify pockets surrounding reactive cysteines suitable for development of small molecule modulators of protein function.

  12. A paper microfluidic cartridge for automated staining of malaria parasites with an optically transparent microscopy window.

    PubMed

    Horning, Matthew P; Delahunt, Charles B; Singh, S Ryan; Garing, Spencer H; Nichols, Kevin P

    2014-06-21

    A paper microfluidic cartridge for the automated staining of malaria parasites (Plasmodium) with acridine orange prior to microscopy is presented. The cartridge enables simultaneous, sub-minute generation of both thin and thick smears of acridine orange stained parasites. Parasites are stained in a cellulose matrix, after which the parasites are ejected via capillary forces into an optically transparent chamber. The unique slanted design of the chamber ensures that a high percentage of the stained blood will be of the required thickness for a thin smear, without resorting to spacers or other methods that can increase production cost or require tight quality controls. A hydrophobic snorkel facilitates the removal of air bubbles during filling. The cartridge contains both a thin smear region, where a single layer of cells is presented unobstructed, for ease of species identification, and a thick smear region, containing multiple cell layers, for enhanced limit of detection. PMID:24781199

  13. Assessing Nezara viridula (Hemiptera: Pentatomidae) feeding damage in macadamia nuts by using a biological stain.

    PubMed

    Golden, Mary; Follett, Peter A; Wright, Mark G

    2006-06-01

    Damage caused by southern green stink bug, Nezara viridula (L.), to macadamia nuts, Macadamia integrifolia Maiden & Betche, is normally determined after nuts are harvested and processed, which may be many months after damage occurred in the field. We developed a method using ruthenium red dye to stain stink bug feeding probes and indirectly assess feeding activity in macadamia nuts. By using the staining method, feeding probes were easily detected on the husk, shell, and kernel. Husk probing was highly correlated (0.80-0.90) with feeding and damage to the kernel. Failure rate to detect kernel damage from stained husk probes was generally <6%. The staining method was equally effective for immature and mature nuts; therefore, N. viridula feeding activity can be monitored throughout the season to evaluate pest management tactics and forecast outbreak populations. PMID:16813317

  14. Efficacy of evaluation of rooster sperm morphology using different staining methods.

    PubMed

    Lukaszewicz, E; Jerysz, A; Partyka, A; Siudzińska, A

    2008-12-01

    This work focused on inexpensive methods of evaluation fowl sperm morphology, based on eosin-nigrosin smears, which can determine disorders in spermatogenesis and can be recommended for evaluating the fertilising potency and selecting males in flocks reproduced by artificial insemination. Four fowl breeds (Black Minorca, Italian Partridge, Forwerk and Greenleg Partridge) were used to determine the efficacy of sperm morphology evaluation using four eosin-nigrosin staining methods (according to Blom, Bakst and Cecil, Morisson, Jaśkowski) and three examiners of different experience (high, medium, novice). There were significant (P< or = 0.01) differences in sperm morphology between Blom's staining method and those of Bakst and Cecil, Morisson or Jaśkowski, irrespective of fowl breed and examiners experience. Blom stain caused sperm head swelling and showed a drastic reduction in the proportion of live spermatozoa with normal morphology. The staining method had a greater influence on sperm morphology evaluation than the experience of the examiners. PMID:18486956

  15. A New Organic Dye-Based Staining for The Detection of Plant DNA in Agarose Gels.

    PubMed

    Sönmezoğlu, Özlem Ateş; Özkay, Kerime

    2015-01-01

    Ethidium bromide (EtBr) is used to stain DNA in agarose gel electrophoresis, but this dye is mutagenic and carcinogenic. We investigated N-719, which is a visible, reliable and organic Ruthenium-based dye, and five fluorescent alternatives for staining plant DNA. For prestaining and poststaining, N-719, GelRed, and SYBR Safe stained both DNA and PCR product bands as clearly as EtBr. SYBR Green I, methylene blue, and crystal violet were effective for poststaining only. The organic dye N-719 stained DNA bands as sensitively and as clearly as EtBr. Consequently, organic dyes can be used as alternatives to EtBr in plant biotechnology studies. PMID:26158569

  16. A staining method for assessing the viability of Esteya vermicola conidia.

    PubMed

    Wang, Yunbo; Thang, NguyenTrong; Li, Zheng; Zhang, Yongan; Li, Jingjie; Xue, Jianjie; Gu, Lijuan; Hong, VuThuy; Mira, Lee; Sung, Changkeun

    2014-07-01

    The viability of conidia of Esteya vermicola, a potentially important biocontrol agent against the pinewood nematode Bursaphelenchus xylophilus, is usually determined by cultivation for 18-48 h in culture medium. As an alternative to this labor-intensive method, we have developed a rapid, simple, and low-cost staining method for assessing E vermicola conidia survival rates. A mixture of neutral red and methylene blue was found to be the most optimal among several stains that also included safranin O and Janus green B. This mixture stained nonviable conidia blue, in contrast to viable conidia, which were stained red in the cytoplasm and blue in the cell wall. This method may be particularly useful for traditional research laboratories, as it provides rapid results using common, relatively inexpensive laboratory equipment. PMID:24585076

  17. Near-Infrared Fluorescence of the NBT/BCIP Chromogenic Stain

    NASA Astrophysics Data System (ADS)

    McCutchen, M. D.; Bumm, L. A.; McCauley, D. W.; Trinh, L. A.; Bonner-Fraser, M.; Fraser, S. E.

    2007-03-01

    We demonstrate the previously unreported near infrared (NIR) fluorescence of the dark purple stain formed from 5-bromo-4-chloro-3-indolyl phosphate (BCIP) and nitro blue tetrazolium (NBT). Although the product is a solid with strong optical absorption, its fluorescence enables high cellular resolution imaging of gene expression. We use spectrofluorometry to identify NBT diformazan as the component of the stain that is the fluorophore exhibiting the strong fluorescence signal. The fluorescence shows an intense emission signal (780-910 nm) that is well separated from excitation (645-685 nm). The NBT diformazan fluorescence is also photostable. Because NBT/BCIP is a widely used chromogenic stain, existing staining protocols can also be applied to fluorescence imaging techniques to increase the resolution of gene expression patterns.

  18. Might the Masson trichrome stain be considered a useful method for categorizing experimental tendon lesions?

    PubMed

    Martinello, Tiziana; Pascoli, Francesco; Caporale, Giovanni; Perazzi, Anna; Iacopetti, Ilaria; Patruno, Marco

    2015-08-01

    Strain injuries of tendons are the most common orthopedic injuries in athletic subjects, be they equine or human. When the tendon is suddenly damaged, an acute inflammatory phase occurs whereas its repetitive overloading may cause chronic injuries. Currently the criteria used for grading injuries are general and subjective, and therefore a reliable grading method would be an improvement. The main purpose of this study was to assess qualitatively the histological pattern of Masson trichrome stain in healthy and injured tendons; indeed, the known "paradox" of Masson staining was used to create an evaluation for the matrix of tendons, following experimental lesions and natural repair processes. A statistically significant difference of aniline-staining between healthy and lesioned tendons was observed. Overall, we think that the Masson staining might be regarded as an informative tool in discerning the collagen spatial arrangement and therefore the histological characteristics of tendons. PMID:25733060

  19. Dual trypan-aniline blue fluorescence staining methods for studying fungus-plant interactions.

    PubMed

    Bhadauria, V; Miraz, P; Kennedy, R; Banniza, S; Wei, Y

    2010-04-01

    Understanding the infection biology of fungi is the key step in devising suitable control strategies for plant diseases. Recently, the Arabidopsis-Colletotrichum higginsianum (causal agent of anthracnose) system has emerged as a seminal paradigm for deciphering the infection biology underlying fungus-plant interactions. We describe here three staining methods coupled with confocal microscopy: trypan blue, aniline blue and dual trypan blue-aniline blue fluorescence staining. Trypan blue and aniline blue staining were employed to scan the infection structures of the hemibiotrophic fungus C. higginsianum and host response in A. thaliana leaf tissues. The two techniques then were combined to observe the contrast between in planta fungal infection structures, i.e., infection vesicles, primary hyphae and secondary hyphae, and the host plant defense responses, i.e., papilla formation and hypersensitive response. These staining techniques also were applied to the lentil-C. truncatum pathosystem to demonstrate their applicability for multiple pathosystems. PMID:19669979

  20. Semiquantitative immunohistochemical marker staining and localization in canine thyroid carcinoma and normal thyroid gland.

    PubMed

    Pessina, P; Castillo, V; Sartore, I; Borrego, J; Meikle, A

    2016-09-01

    Immunoreactive proteins in follicular cells, fibroblasts and endothelial cells were assessed in canine thyroid carcinomas and healthy thyroid glands. No differences were detected in thyrotropin receptor and thyroglobulin staining between cancer and normal tissues, but expression was higher in follicular cells than in fibroblasts. Fibroblast growth factor-2 staining was more intense in healthy follicular cells than in those of carcinomas. Follicular cells in carcinomas presented two- to three-fold greater staining intensity of thyroid transcription factor-1 and proliferating cell nuclear antigen, respectively, than healthy cells, and a similar trend was found for the latter antigen in fibroblasts. Vascular endothelial growth factor staining was more intense in the endothelial cells of tumours than in those of normal tissues. In conclusion, greater expression of factors related to proliferation and angiogenesis was demonstrated in several cell types within thyroid carcinomas compared to healthy tissues, which may represent mechanisms of tumour progression in this disease. PMID:25082554