Sample records for conditioned medium prepared

  1. Investigation of mitomycin-C-treated fibroblasts in 3-D collagen gel and conditioned medium for keratinocyte proliferation.

    PubMed

    Huang, Yi-Chau; Wang, Tzu-Wei; Sun, Jui-Sheng; Lin, Feng-Huei

    2006-03-01

    Fibroblasts produce a spectrum of necessary growth factors essential for growth and proliferation of a variety of cell types. In this study, the paracrine effect of mitomycin-C-treated fibroblasts with various densities in collagen gel for keratinocyte proliferation was investigated from which an optimum cell density and optimum conditioned medium would be determined to expand keratinocyte without further differentiation for skin equivalent tissue engineering. The optimum cell density in collagen feeder gel for optimum collected medium preparation will be determined by checking the level of keratinocyte growth factor and granulocyte macrophage colony-stimulating factor in conventional medium. The results showed that the cell density of 1 x 10(5) cells/gel in the feeder gel is better to produce optimum collected medium. The conditioned medium is prepared by mixing together the optimum collected medium and molecular cellular and developmental biology (MCDB) 153 medium in different ratios for keratinocyte growth. The keratinocyte viability will be measured by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to determine the optimum conditioned medium. From the study, 67% conditioned medium was supposed as the better medium for keratinocyte proliferation. In this experiment, the optimum cell density in feeder gel to coculture with keratinocytes is also determined as 1 x 10(5) cells/gel. Keratin 10 (K10) and Terminal Deoxynucleotidyl Transferase Mediated dUTP Nick End Labeling stain will be used to check the cell differentiation and apoptosis, respectively. The results suggest that keratinocytes should not be cultured in postconfluent conditions due to undesired apoptosis and differentiation. The result of cell viability from passages to passages shows that the optimum feeder gel plays a more important role to the keratinocyte proliferation than that of optimum conditioned medium. Keratinocytes cultured with optimum feeder gel in 67% conditioned medium could effectively promote proliferation, inhibit apoptosis, and prevent differentiation. The combination of conditioned media and feeder gel to culture keratinocytes without external supplements can provide an inexpensive way for keratinocyte proliferation and construct an environment for real-time communication between the two cells. The results conclude that keratinocyte cultivation in feeder gel with modified medium should be feasible in the production of high quality keratinocytes for skin equivalents preparation.

  2. Preparing the Production of a New Product in Small and Medium-Sized Enterprises by Using the Method of Projects Management

    NASA Astrophysics Data System (ADS)

    Bijańska, Jolanta; Wodarski, Krzysztof; Wójcik, Janusz

    2016-06-01

    Efficient and effective preparation the production of new products is important requirement for a functioning and development of small and medium-sized enterprises. One of the methods, which support the fulfilment of this condition is project management. This publication presents the results of considerations, which are aimed at developing a project management model of preparation the production of a new product, adopted to specificity of small and medium-sized enterprises.

  3. Thermal processing of bone: in vitro response of mesenchymal cells to bone-conditioned medium.

    PubMed

    Sawada, K; Caballé-Serrano, J; Schuldt Filho, G; Bosshardt, D D; Schaller, B; Buser, D; Gruber, R

    2015-08-01

    The autoclaving, pasteurization, and freezing of bone grafts to remove bacteria and viruses, and for preservation, respectively, is considered to alter biological properties during graft consolidation. Fresh bone grafts release paracrine-like signals that are considered to support tissue regeneration. However, the impact of the autoclaving, pasteurization, and freezing of bone grafts on paracrine signals remains unknown. Therefore, conditioned medium was prepared from porcine cortical bone chips that had undergone thermal processing. The biological properties of the bone-conditioned medium were assessed by examining the changes in expression of target genes in oral fibroblasts. The data showed that conditioned medium obtained from bone chips that had undergone pasteurization and freezing changed the expression of adrenomedullin, pentraxin 3, BTB/POZ domain-containing protein 11, interleukin 11, NADPH oxidase 4, and proteoglycan 4 by at least five-fold in oral fibroblasts. Bone-conditioned medium obtained from autoclaved bone chips, however, failed to change the expression of the respective genes. Also, when bone-conditioned medium was prepared from fresh bone chips, autoclaving blocked the capacity of bone-conditioned medium to modulate gene expression. These in vitro results suggest that pasteurization and freezing of bone grafts preserve the release of biologically active paracrine signals, but autoclaving does not. Copyright © 2015 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  4. The in vitro impact of toothpaste extracts on cell viability.

    PubMed

    Cvikl, Barbara; Lussi, Adrian; Gruber, Reinhard

    2015-06-01

    Toothpastes contain three main components: detergents, abrasives, and fluoride. Detergents, particularly sodium lauryl sulfate, have been proposed as components that enable toothpastes to produce cytotoxic effects in vitro. However, not all toothpastes contain sodium lauryl sulfate, and almost no studies have found an association between detergents and the in vitro cytotoxicity of toothpastes. The present study examined the in vitro cytotoxicity of nine commercially available toothpastes containing four different detergents. Toothpastes were diluted in serum-free medium, centrifuged, and filter sterilized. The half-lethal concentration of the toothpaste-conditioned medium (TCM) was calculated based on the formation of formazan by gingival fibroblasts, oral squamous cell carcinoma HSC-2 cells, and L929 cells. Cell proliferation was analyzed, and live-dead staining was performed, after exposure of cells to conditioned medium prepared with 1% toothpaste (1% TCM). It was found that toothpastes containing sodium lauryl sulfate and amine fluoride strongly inhibited cell viability with the half-lethal concentration being obtained with conditioned medium prepared with approximately 1% toothpaste (1% TCM). Toothpastes containing cocamidopropyl betaine and Steareth-20 showed higher half-lethal concentration values, with the half-lethal concentration being obtained with conditioned medium prepared with 10% (10% TCM) and 70% (70% TCM) toothpaste, respectively. Proliferation and live-dead data were consistent with the cell-viability analyses. These results demonstrate that the type of detergent in toothpastes can be associated with changes in in vitro cell toxicity. © 2015 Eur J Oral Sci.

  5. In Vitro Impact of Conditioned Medium From Demineralized Freeze-Dried Bone on Human Umbilical Endothelial Cells.

    PubMed

    Harnik, Branko; Miron, Richard J; Buser, Daniel; Gruber, Reinhard

    2017-03-01

    Angiogenesis is essential for the consolidation of bone allografts. The underlying molecular mechanism, however, remains unclear. Soluble factors released from demineralized freeze-dried bone target mesenchymal cells; however, their effect on endothelial cells has not been investigated so far. The aim of the present study was therefore to examine the effect of conditioned medium from demineralized freeze-dried bone on human umbilical endothelial cells in vitro. Conditioned medium was first prepared from demineralized freeze-dried bone following 24 hours incubation at room temperature to produce demineralized bone conditioned media. Thereafter, conditioned medium was used to stimulate human umbilical vein endothelial cells in vitro by determining the cell response based on viability, proliferation, expression of apoptotic genes, a Boyden chamber to determine cell migration, and the formation of branches. The authors report here that conditioned medium decreased viability and proliferation of endothelial cells. Neither of the apoptotic marker genes was significantly altered when endothelial cells were exposed to conditioned medium. The Boyden chamber revealed that endothelial cells migrate toward conditioned medium. Moreover, conditioned medium moderately stimulated the formation of branches. These findings support the concept that conditioned medium from demineralized freeze-dried bone targets endothelial cells by decreasing their proliferation and enhancing their motility under these in vitro conditions.

  6. Green materials prepared by using a green processing method: Plant oil-based polymers prepared in CO2 medium with mild conditions

    USDA-ARS?s Scientific Manuscript database

    Recent efforts have been focused on the development of environmentally friendly replacements for volatile organic solvents. One promising candidate is supercritical carbon dioxide because of its low toxicity and no solvent residues in the final products. Renewed interest in biodegradable polymeric m...

  7. Detection of gain enhancement in laser-induced fluorescence of rhodamine B lasing dye by silicon dioxide nanostructures-coated cavity

    NASA Astrophysics Data System (ADS)

    Al-Tameemi, Mohammed N. A.

    2018-03-01

    In this work, nanostructured silicon dioxide films are deposited by closed-field unbalanced direct-current (DC) reactive magnetron sputtering technique on two sides of quartz cells containing rhodamine B dye dissolved in ethanol with 10‒5 M concentration as a random gain medium. The preparation conditions are optimized to prepare highly pure SiO2 nanostructures with a minimum particle size of about 20 nm. The effect of SiO2 films as external cavity for the random gain medium is determined by the laser-induced fluorescence of this medium, and an increase of about 200% in intensity is observed after the deposition of nanostructured SiO2 thin films on two sides of the dye cell.

  8. Development of conditioning programs for dressage horses based on time-motion analysis of competitions.

    PubMed

    Clayton, H M

    1993-05-01

    The time-motion characteristics of Canadian basic- and medium-level dressage competitions are described, and the results are applied in formulating sport-specific conditioning programs. One competition was analyzed at the six levels from basic 1 to medium 3. Each test was divided into a series of sequences based on the type and speed of activity. The durations of the sequences were measured from videotapes. The basic-level tests had fewer sequences, and they were shorter in distance and duration than the medium tests (P < 0.10), but the average speed did not differ between the two levels. It is recommended that horses competing at the basic levels be conditioned using 5-min exercise periods, with short (10-s) bursts of lengthened trot and canter included at basic 2 and above. In preparation for medium-level competitions, the duration of the work periods increases to 7 min, 10- to 12-s bursts of medium or extended trot and canter are included, and transitions are performed frequently to simulate the energy expenditure in overcoming inertia.

  9. A novel composite material based on antimony(III) oxide and amorphous silica

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

    Zemnukhova, Ludmila A.; Panasenko, Alexander E., E-mail: panasenko@ich.dvo.ru

    2013-05-01

    The composite material nSb₂O₃·mSiO₂·xH₂O was prepared by hydrolysis of SbCl₃ and Na₂SiO₃ in an aqueous medium. It has been shown that the composition of the material is influenced by the ratio of the initial components and the acidity of the reaction medium. The morphology of the material particles and its specific surface area have been determined. The thermal and optic properties were also investigated. - Graphical abstract: Novel composite material containing amorphous silica and crystal antimony(III) oxide has been synthesized by hydrolysis of SbCl₃ and Na₂SiO₃ in an aqueous medium. Highlights: • The composite material nSb₂O₃·mSiO₂·xH₂O was prepared in anmore » aqueous medium. • The composition of the material is controllable by a synthesis conditions. • The morphology of the material and its optic properties have been determined.« less

  10. The role of hydrothermally prepared supported photocatalytic composite in organic micro-pollutants removal from the water.

    PubMed

    Shivaraju, H P; Byrappa, K

    2012-07-01

    This work deals with the non-biodegradable micro-pollutants degradation by supported photocatalyst based heterogeneous photocatalytic reaction. TiO2 based supported photocatalyst was prepared by the hydrothermal technique to improve the photocatalytic performance along with easy recovery of suspended photocatalyst from aqueous medium after photoreaction. TO2 deposited calcium alumino-silicate beads (CASB) supports were prepared under mild hydrothermal conditions (Temparature-200 degrees C, Duration-24 h). In the present study, industrial dyes such as Amaranth and Brilliant Yellow were used as model micro-pollutants in aqueous solution. A real time pesticide industrial effluent was tested for its photocatalytic removal of organic pollutants using TO2 deposited CASB supported photocatalytic composite as an effective photocatalyst. Photocatalytic degradation of micro-pollutants present in aqueous medium was carried out in a batch photoreactor, at atmospheric pressure and temperature (28 degrees C). The influence of different light sources, irradiation time, catalyst load and catalytic performance is discussed. The photocatalytic degradation of micro-pollutants in aqueous medium was evaluated by determination of COD and %T. Easy separation and recovery of suspended photocatalysts from aqueous solution is the major advantage of hydrothermally prepared supported photocatalytic composite.

  11. Study of cellulolytic soil fungi and two nova species and new medium

    PubMed Central

    Khalid, Mahmood; Yang, Wei-jun; Kishwar, Nazir; Rajput, Zahid Iqbal; Arijo, Abdullah G.

    2006-01-01

    This study is aimed at identifying and determining the percentage of occurrence frequency of cellulose decomposing soil fungi. The soil samples were inoculated into culture plates prepared in Sabouraud medium under sterilized conditions and incubated at 30 °C for 4 to 7 d. The identified fungal species were incubated in self-designed cellulose medium for testing their cellulolytic ability. Forty-two species, including 2 nova species, representing sixteen genera showed growth and sporulation in the cellulose medium. Most of the isolated species were from genus Aspergillus and Penicillium. Aspergillus niger and Mucor hiemalis showed highest occurrence frequency (45% and 36% respectively), as these species were collected from about 80% of soil samples. Being agar free and cheaper, the new fungal medium designed showed results equivalent to Sabouraud medium. PMID:16691640

  12. Characterization of Residual Medium Peptides from Yersinia pestis Cultures

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

    Clowers, Brian H.; Wunschel, David S.; Kreuzer, Helen W.

    2013-04-03

    Using a range of common microbial medium formulations (TSB, BHI, LB, and G-media), two attenuated strains of Y. pestis (KIM D27 (pgm-) and KIMD1 lcr-) were cultivated in triplicate. These cellular suspensions were used to develop a method of extracting residual medium peptides from the final microbial preparation to assess their relative abundance and identity. Across the conditions examined, which included additional cellular washing and different forms of microbial inactivation, residual medium peptides were detected. Despite the range of growth medium sources used and the associated manufacturing processes used in their production, a high degree of peptide similarity was observedmore » for a given medium recipe. These results demonstrate that residual medium peptides are retained using traditional microbial cultivation techniques and may be used to inform forensic investigations with respect to production deduction.« less

  13. Method for loading lipsomes with ionizable phosphorylated hydrophobic compounds, pharmaceutical preparations and a method for administering the preparations

    DOEpatents

    Mehlhorn, Rolf Joachim

    1998-10-27

    A method of entrapping ionizable compounds, preferably phosphorylated hydrophobic compounds, into liposomes having transmembrane gradients is disclosed. The procedures involve forming liposomes in an acidic medium or a basic medium, adding to the acidic medium a cationic compound or to the basic medium an anionic compound and then adding a base to the cationic-containing medium or an acid to the anionic-containing medium, thereby inducing the ionizable compound into the liposomes' internal aqueous phase. The compound-entrapped liposomes prepared in accordance with the disclosed methods may be used as pharmaceutical preparations. Methods of administering such pharmaceutical preparations are also disclosed.

  14. Isolation of naturally infecting Leishmania infantum from canine samples in Novy-MacNeal-Nicolle medium prepared with defibrinated blood from different animal species.

    PubMed

    Santos, Roseclea Chagas Dos; Pinho, Flaviane Alves de; Passos, Gabriela Porfírio; Larangeira, Daniela Farias; Barrouin-Melo, Stella Maria

    2018-06-15

    The most commonly used culture medium for the in vitro isolation of Leishmania spp. from canine biological samples is biphasic Novy-MacNeal-Nicolle (NNN) medium, whose solid phase is prepared using rabbit blood. Leishmania infantum parasites from natural infections are highly sensitive and demanding for growth in axenic conditions when firstly obtained from the dog's body. The objective of this study was to evaluate whether NNN medium (NNN-test) prepared with chicken blood (NNN-C), ox blood (NNN-O), horse blood (NNN-H) or sheep blood (NNN-S) was viable for the isolation of parasites from naturally infected dogs, in an endemic area for visceral leishmaniasis caused by L. infantum. Spleen aspirates from six dogs previously diagnosed as infected by parasitological methods were simultaneously inoculated in each NNN-test medium, including the conventional medium prepared with rabbit blood (NNN-R), and the cultures were examined for three weeks under optic microscopy. Spleen samples were also analyzed for parasite loads by quantitative PCR (qPCR). Cultures from three of the six dogs (50%) were positive in at least one of the NNN-test media: one sample presented the highest spleen parasite load by qPCR (1.19 × 10 4 parasites/mL) and was positive in all test media; the second sample presented parasitic isolation in the first week of culture in all inoculated media, of which the NNN-C medium had the highest mean parasite count (NNN-C = 23.5 × 10 4 /mL vs. NNN-R = 3.25 × 10 4 /mL); the third sample was positive only in the NNN-S medium besides the conventional control NNN-R. Cultures from the three remaining dogs were negative in all NNN media, including the control and test media; of those three dogs, two presented the lowest spleen parasitic loads according to qPCR. Blood from chicken, ox, horse and sheep shown to be viable for the preparation of NNN culture medium for the primary isolation of L. infantum from samples of naturally infected dogs and can be considered as an alternative to rabbit blood when necessary. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Nanosized UCMSC-derived extracellular vesicles but not conditioned medium exclusively inhibit the inflammatory response of stimulated T cells: implications for nanomedicine.

    PubMed

    Monguió-Tortajada, Marta; Roura, Santiago; Gálvez-Montón, Carolina; Pujal, Josep Maria; Aran, Gemma; Sanjurjo, Lucía; Franquesa, Marcel la; Sarrias, Maria-Rosa; Bayes-Genis, Antoni; Borràs, Francesc E

    2017-01-01

    Undesired immune responses have drastically hampered outcomes after allogeneic organ transplantation and cell therapy, and also lead to inflammatory diseases and autoimmunity. Umbilical cord mesenchymal stem cells (UCMSCs) have powerful regenerative and immunomodulatory potential, and their secreted extracellular vesicles (EVs) are envisaged as a promising natural source of nanoparticles to increase outcomes in organ transplantation and control inflammatory diseases. However, poor EV preparations containing highly-abundant soluble proteins may mask genuine vesicular-associated functions and provide misleading data. Here, we used Size-Exclusion Chromatography (SEC) to successfully isolate EVs from UCMSCs-conditioned medium. These vesicles were defined as positive for CD9, CD63, CD73 and CD90, and their size and morphology characterized by NTA and cryo-EM. Their immunomodulatory potential was determined in polyclonal T cell proliferation assays, analysis of cytokine profiles and in the skewing of monocyte polarization. In sharp contrast to the non-EV containing fractions, to the complete conditioned medium and to ultracentrifuged pellet, SEC-purified EVs from UCMSCs inhibited T cell proliferation, resembling the effect of parental UCMSCs. Moreover, while SEC-EVs did not induce cytokine response, the non-EV fractions, conditioned medium and ultracentrifuged pellet promoted the secretion of pro-inflammatory cytokines by polyclonally stimulated T cells and supported Th17 polarization. In contrast, EVs did not induce monocyte polarization, but the non-EV fraction induced CD163 and CD206 expression and TNF-α production in monocytes. These findings increase the growing evidence confirming that EVs are an active component of MSC's paracrine immunosuppressive function and affirm their potential for therapeutics in nanomedicine. In addition, our results highlight the importance of well-purified and defined preparations of MSC-derived EVs to achieve the immunosuppressive effect.

  16. wnt3a but not wnt11 supports self-renewal of embryonic stem cells

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

    Singla, Dinender K.; Schneider, David J.; LeWinter, Martin M.

    2006-06-30

    wnt proteins (wnts) promote both differentiation of midbrain dopaminergic cells and self-renewal of haematopoietic stem cells. Mouse embryonic stem (ES) cells can be maintained and self-renew on mouse feeder cell layers or in media containing leukemia inhibitory factor (LIF). However, the effects of wnts on ES cells self-renewal and differentiation are not clearly understood. In the present study, we found that conditioned medium prepared from L cells expressing wnt3a can replace feeder cell layers and medium containing LIF in maintaining ES cells in the proliferation without differentiation (self-renewal) state. By contrast, conditioned medium from NIH3T3 cells expressing wnt11 did not.more » Alkaline phosphatase staining and compact colony formation were used as criteria of cells being in the undifferentiated state. ES cells maintained in medium conditioned by Wnt3a expressing cells underwent freezing and thawing while maintaining properties seen with LIF maintained ES cells. Purified wnt3a did not maintain self-renewal of ES cells for prolonged intervals. Thus, other factors in the medium conditioned by wnt3a expressing cells may have contributed to maintenance of ES cells in a self-renewal state. Pluripotency of ES cells was determined with the use of embryoid bodies in vitro. PD98059, a MEK specific inhibitor, promoted the growth of undifferentiated ES cells maintained in conditioned medium from wnt3a expressing cells. By contrast, the P38 MAPK inhibitor SB230580 did not, suggesting a role for the MEK pathway in self-renewal and differentiation of ES cells maintained in the wnt3a cell conditioned medium. Thus, our results show that conditioned medium from wnt3a but not wnt11 expressing cells can maintain ES cells in self-renewal and in a pluripotent state.« less

  17. Method for loading lipsomes with ionizable phosphorylated hydrophobic compounds, pharmaceutical preparations and a method for administering the preparations

    DOEpatents

    Mehlhorn, R.J.

    1998-10-27

    A method of entrapping ionizable compounds, preferably phosphorylated hydrophobic compounds, into liposomes having transmembrane gradients is disclosed. The procedures involve forming liposomes in an acidic medium or a basic medium, adding to the acidic medium a cationic compound or to the basic medium an anionic compound and then adding a base to the cationic-containing medium or an acid to the anionic-containing medium, thereby inducing the ionizable compound into the liposomes` internal aqueous phase. The compound-entrapped liposomes prepared in accordance with the disclosed methods may be used as pharmaceutical preparations. Methods of administering such pharmaceutical preparations are also disclosed. 2 figs.

  18. Kinetics of oil saponification by lead salts in ancient preparations of pharmaceutical lead plasters and painting lead mediums.

    PubMed

    Cotte, M; Checroun, E; Susini, J; Dumas, P; Tchoreloff, P; Besnard, M; Walter, Ph

    2006-12-15

    Lead soaps can be found in archaeological cosmetics as well as in oil paintings, as product of interactions of lead salts with oil. In this context, a better understanding of the formation of lead soaps allows a follow-up of the historical evolution of preparation recipes and provides new insights into conservation conditions. First, ancient recipes of both pharmaceutical lead plasters and painting lead mediums, mixtures of oil and lead salts, were reconstructed. The ester saponification by lead salts is determined by the preparation parameters which were quantified by FT-IR spectrometry. In particular, ATR/FT-IR spectrometer was calibrated by the standard addition method to quantitatively follow the kinetics of this reaction. The influence of different parameters such as temperature, presence of water and choice of lead salts was assessed: the saponification is clearly accelerated by water and heating. This analysis provides chemical explanations to the historical evolution of cosmetic and painting preparation recipes.

  19. Conditioned medium of adipose-derived stromal cell culture in three-dimensional bioreactors for enhanced wound healing.

    PubMed

    Kwon, Sun Hyun; Bhang, Suk Ho; Jang, Hyeon-Ki; Rhim, Taiyoun; Kim, Byung-Soo

    2015-03-01

    It was previously shown that human adipose-derived stromal cell (hADSC)-conditioned medium (CM) promotes wound healing. An essential part of the wound healing process is neovascularization in the wound bed. We hypothesized that CM prepared from hADSCs cultured as spheroids in three-dimensional suspension bioreactors (spheroid CM) would contain much higher concentrations of angiogenic growth factors secreted by hADSCs, induce a higher extent of neovascularization in the wound bed, and improve wound healing as compared with CM prepared by conventional monolayer culture (monolayer CM). The concentrations of angiogenic growth factors (i.e., vascular endothelial growth factor, basic fibroblast growth factor, and hepatocyte growth factor) in spheroid CM were 20- to 145-fold higher than those in monolayer CM. Either fresh medium, monolayer CM, or spheroid CM was administered to full-thickness wounds created on the dorsal aspects of athymic mice. The monolayer CM promoted wound healing as compared with fresh medium or no treatment. Importantly, wound closure was faster, and dermal and epidermal regeneration was improved in the spheroid CM-treated mice compared with that in the monolayer CM-treated mice. The improved wound healing by spheroid CM may be attributed, at least in part, to enhanced neovascularization in the wound beds. The spheroid-based CM approach showed potential as a therapy for skin wound repair. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Effect of medium variations (zinc supplementation during oocyte maturation, perifertilization pH, and embryo culture protein source) on equine embryo development after intracytoplasmic sperm injection.

    PubMed

    Choi, Young-Ho; Gibbons, John R; Canesin, Heloísa S; Hinrichs, Katrin

    2016-10-15

    Prospective studies were conducted to help define procedural factors affecting in vitro embryo production via intracytoplasmic sperm injection (ICSI) of equine oocytes. In experiment 1, use of 10% fetal bovine serum as a protein source in embryo culture medium resulted in a higher blastocyst rate than did use of a combination of 3% fetal bovine serum, 3% equine preovulatory follicular fluid, and 4% human serum substitute (37% vs. 15%, respectively, P < 0.05). In experiment 2, the effect of zinc supplementation (0, 0.5, 1, or 1.5 μg/mL) during IVM was examined. There were no significant differences in rates of cleavage or blastocyst development (20%-31%). However, the proportion of blastocysts that developed on Day 7 for the added-zinc treatments was significantly higher than that for the control treatment (45% vs. 8%). In experiment 3, we tested whether use of high-pH medium (pH 8.0-8.4) during ICSI procedures would improve blastocyst rate when sperm with low cleavage rates after ICSI was used. When high-pH conditions were used for sperm preparation and also for the first 2 hours of incubation of injected oocytes after ICSI, the cleavage rate was unaffected but no blastocysts developed (0% vs. 24% for control). When high-pH conditions were used for sperm preparation only, the blastocyst rate was 37%. This was repeated using sperm from a second stallion; there was no significant difference in cleavage or blastocyst rates between sperm preparation in high pH vs. control medium. These findings add to our knowledge of factors affecting in vitro production of equine embryos. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. [Preparation of bioindicators for controlling the efficacy of sterilization processes].

    PubMed

    Kalinina, N M; Tikhonova, A S; Motina, G L; Chaĭkovskaia, S M; Semenov, S M

    1983-08-01

    A test microbe for the control of the efficacy of vacuum steam sterilization was selected. Conditions for the spore cultivation were developed. The optimal bioindicator composition, test microbe loading, nutrient medium, pH indicator, carbohydrates and a carrier were defined.

  2. Effects of neural stem cell media on hypoxic injury in rat hippocampal slice cultures.

    PubMed

    Lee, Na Mi; Chae, Soo Ahn; Lee, Hong Jun

    2017-12-15

    Neonatal hypoxic-ischemic brain injuries cause serious neurological sequelae, yet there is currently no effective treatment for them. We hypothesized that neurotrophic factors released into the medium by stem cells could supply hypoxia-damaged organotypic hippocampal slice cultures with regenerative abilities. We prepared organotypic slice cultures of the hippocampus of 7-day-old Sprague-Dawley rats based on the modified Stoppini method; slices were cultured for 14days in vitro using either Gahwiler's medium (G-medium) or stem cell-conditioned medium (S-medium) as culture medium. At day 14 in vitro, hippocampal slice cultures were exposed to 95% N 2 and 5% CO 2 for 3h to induce hypoxic damage, the extent of which was then measured using propidium iodide fluorescence and immunohistochemistry images. We performed dot blotting to estimate neurotrophic/growth factor levels in the G- and S-media. Organotypic hippocampal slices cultured using S-medium after hypoxic injury were significantly less damaged than those cultured using G-medium. GLUT1, NGF, GDNF, VEGF, GCSF, and IGF2 levels were higher in S-medium than in G-medium, whereas FGF1, HIF, and MCP3 levels were not significantly different between media. In conclusion, we found that stem cell-conditioned medium had a neuroprotective effect against hypoxic injury, and that, of the various neurotrophic factors in S-medium, NGF, GDNF, and VEGF can contribute to neuroprotection. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Preparation of ionic-crosslinked chitosan-based gel beads and effect of reaction conditions on drug release behaviors.

    PubMed

    Chen, Shilan; Liu, Mingzhu; Jin, Shuping; Wang, Bin

    2008-02-12

    Drug-loaded chitosan (CS) beads were prepared under simple and mild condition using trisodium citrate as ionic crosslinker. The beads were further coated with poly(methacrylic acid) (PMAA) by dipping the beads in PMAA aqueous solution. The surface and cross-section morphology of these beads were observed by scanning electron microscopy and the observation showed that the coating beads had core-shell structure. In vitro release of model drug from these beads obtained under different reaction conditions was investigated in buffer medium (pH 1.8). The results showed that the rapid drug release was restrained by PMAA coating and the optimum conditions for preparing CS-based drug-loaded beads were decided through the effect of reaction conditions on the drug release behaviors. In addition, the drug release mechanism of CS-based drug-loaded beads was analyzed by Peppa's potential equation. According to this study, the ionic-crosslinked CS beads coated by PMAA could serve as suitable candidate for drug site-specific carrier in stomach.

  4. Metal glass vacuum tube solar collectors are approaching lower-medium temperature heat application.

    PubMed

    Jiang, Xinian

    2010-04-26

    Solar thermal collectors are widely used worldwide mainly for hot water preparation at a low temperature (less than 80?C). Applications including many industrial processes and central air conditioning with absorption chillers, instead require lower-medium temperature heat (between 90 degrees C and 150 degrees C) to be driven when using solar thermal energy. The metal absorber glass vacuum tube collectors (MGVT) are developed for this type of applications. Current state-of-art and possible future technology development of MGVT are presented.

  5. Using straw hydrolysate to cultivate Chlorella pyrenoidosa for high-value biomass production and the nitrogen regulation for biomass composition.

    PubMed

    Zhang, Tian-Yuan; Wang, Xiao-Xiong; Wu, Yin-Hu; Wang, Jing-Han; Deantes-Espinosa, Victor M; Zhuang, Lin-Lan; Hu, Hong-Ying; Wu, Guang-Xue

    2017-11-01

    Heterotrophic cultivation of Chlorella pyrenoidosa based on straw substrate was proposed as a promising approach in this research. The straw pre-treated by ammonium sulfite method was enzymatically hydrolyzed for medium preparation. The highest intrinsic growth rate of C. pyrenoidosa reached to 0.097h -1 in hydrolysate medium, which was quicker than that in glucose medium. Rising nitrogen concentration could significantly increase protein content and decrease lipid content in biomass, meanwhile fatty acids composition kept stable. The highest protein and lipid content in microalgal biomass reached to 62% and 32% under nitrogen excessive and deficient conditions, respectively. Over 40% of amino acids and fatty acids in biomass belonged to essential amino acids (EAA) and essential fatty acids (EFA), which were qualified for high-value uses. This research revealed the rapid biomass accumulation property of C. pyrenoidosa in straw hydrolysate medium and the effectiveness of nitrogen regulation to biomass composition at heterotrophic condition. Copyright © 2017. Published by Elsevier Ltd.

  6. Viability of BCG suspensions, freshly prepared, stored, and light-exposed, estimated in different ways.

    PubMed

    CHRISTENSEN, P A; ROBINSON, M; WIDDICOMBE, M

    1955-01-01

    Tested by the roll-tube method, the inclusion in Dubos medium of oleic acid, Tween 80, or glucose hastens colony growth of tubercle bacilli (BCG), but the variability in counts between replicate bottles is large, and the mean count is low compared with that obtained in media free of these substances.The addition of glycerol hastens the development of colonies, and counts on glycerol medium may differ from those on glycerol-free medium. BCG suspensions stored at about 23 degrees C or exposed to skyshine or sunlight become glycerol-sensitive. Results obtained with glycerol medium may not, therefore, always be acceptable.The preparation and use in the roll-tube method of a simple medium is described. This consists of horse serum, M/15 phosphate buffer, and agar, and is preferable to more complex media as it tends to give higher viable counts and is easier to store and prepare.Stored at about 23 degrees C, the viability of BCG is better preserved in neutral phosphate buffer than in suspending fluids containing Sauton medium; no such difference is noticed with cold storage.Glutamic acid added in a concentration of 0.35% is without effect on the viability of suspensions stored in the cold, but under certain conditions it may have some preserving value at higher storage temperatures.Exposure to daylight in the laboratory, even for several hours, does not kill BCG or render it glycerol-sensitive. Exposure to intense skyshine does kill, but the mortality observed at the South African Institute for Medical Research is low compared with that recorded elsewhere. Possible explanations of this discrepancy are discussed.

  7. Viability of BCG suspensions, freshly prepared, stored, and light-exposed, estimated in different ways

    PubMed Central

    Christensen, P. Agerholm; Robinson, Mary; Widdicombe, Margaret

    1955-01-01

    Tested by the roll-tube method, the inclusion in Dubos medium of oleic acid, Tween 80, or glucose hastens colony growth of tubercle bacilli (BCG), but the variability in counts between replicate bottles is large, and the mean count is low compared with that obtained in media free of these substances. The addition of glycerol hastens the development of colonies, and counts on glycerol medium may differ from those on glycerol-free medium. BCG suspensions stored at about 23°C or exposed to skyshine or sunlight become glycerol-sensitive. Results obtained with glycerol medium may not, therefore, always be acceptable. The preparation and use in the roll-tube method of a simple medium is described. This consists of horse serum, M/15 phosphate buffer, and agar, and is preferable to more complex media as it tends to give higher viable counts and is easier to store and prepare. Stored at about 23°C, the viability of BCG is better preserved in neutral phosphate buffer than in suspending fluids containing Sauton medium; no such difference is noticed with cold storage. Glutamic acid added in a concentration of 0.35% is without effect on the viability of suspensions stored in the cold, but under certain conditions it may have some preserving value at higher storage temperatures. Exposure to daylight in the laboratory, even for several hours, does not kill BCG or render it glycerol-sensitive. Exposure to intense skyshine does kill, but the mortality observed at the South African Institute for Medical Research is low compared with that recorded elsewhere. Possible explanations of this discrepancy are discussed. PMID:14379008

  8. l-Asparaginase from Proteus vulgaris1

    PubMed Central

    Tosa, Tetsuya; Sano, Ryujiro; Yamamoto, Kozo; Nakamura, Masatoshi; Ando, Katsuko; Chibata, Ichiro

    1971-01-01

    To produce an immunologically and enzymologically new type of l-asparaginase, 108 strains of bacteria were screened for enzyme production. As a result, 13 bacteria belonging to the genera Alcaligenes, Bacterium, and Proteus were found to produce l-asparaginases in high levels. Among these l-asparaginases, partially purified l-asparaginases from B. cadaveris and P. vulgaris showed antitumor activity. A partially purified l-asparaginase preparation of P. vulgaris did not react with the antibody of Escherichia colil-asparaginase on the Ouchterlony agar plate. Culture conditions for the production of l-asparaginase by P. vulgaris were investigated in detail. The enzyme was produced in high yields when cells were grown aerobically in a medium containing sodium fumarate and corn steep liquor. The addition of glucose or ammonium ion to the medium, however, resulted in depressed production of l-asparaginase. Under the optimum conditions, 3,700 international units of l-asparaginase was obtained from 1 liter of culture medium. Images PMID:5000866

  9. Nanoparticle growth and surface chemistry changes in cell-conditioned culture medium.

    PubMed

    Kendall, Michaela; Hodges, Nikolas J; Whitwell, Harry; Tyrrell, Jess; Cangul, Hakan

    2015-02-05

    When biomolecules attach to engineered nanoparticle (ENP) surfaces, they confer the particles with a new biological identity. Physical format may also radically alter, changing ENP stability and agglomeration state within seconds. In order to measure which biomolecules are associated with early ENP growth, we studied ENPs in conditioned medium from A549 cell culture, using dynamic light scattering (DLS) and linear trap quadrupole electron transfer dissociation mass spectrometry. Two types of 100 nm polystyrene particles (one uncoated and one with an amine functionalized surface) were used to measure the influence of surface type. In identically prepared conditioned medium, agglomeration was visible in all samples after 1 h, but was variable, indicating inter-sample variability in secretion rates and extracellular medium conditions. In samples conditioned for 1 h or more, ENP agglomeration rates varied significantly. Agglomerate size measured by DLS was well correlated with surface sequestered peptide number for uncoated but not for amine coated polystyrene ENPs. Amine-coated ENPs grew much faster and into larger agglomerates associated with fewer sequestered peptides, but including significant sequestered lactose dehydrogenase. We conclude that interference with extracellular peptide balance and oxidoreductase activity via sequestration is worthy of further study, as increased oxidative stress via this new mechanism may be important for cell toxicity. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

  10. SnAP reagents for the one-step synthesis of medium-ring saturated N-heterocycles from aldehydes

    NASA Astrophysics Data System (ADS)

    Vo, Cam-Van T.; Luescher, Michael U.; Bode, Jeffrey W.

    2014-04-01

    Interest in saturated N-heterocycles as scaffolds for the synthesis of bioactive molecules is increasing. Reliable and predictable synthetic methods for the preparation of these compounds, especially medium-sized rings, are limited. We describe the development of SnAP (Sn amino protocol) reagents for the transformation of aldehydes into seven-, eight- and nine-membered saturated N-heterocycles. This process occurs under mild, room-temperature conditions and offers exceptional substrate scope and functional-group tolerance. Air- and moisture-stable SnAP reagents are prepared on a multigram scale from inexpensive starting materials by simple reaction sequences. These new reagents and processes allow widely available aryl, heteroaryl and aliphatic aldehydes to be converted into diverse N-heterocycles, including diazepanes, oxazepanes, diazocanes, oxazocanes and hexahydrobenzoxazonines, by a single synthetic operation.

  11. Preparation time influences ankle and knee joint control during dynamic change of direction movements.

    PubMed

    Fuerst, Patrick; Gollhofer, Albert; Gehring, Dominic

    2017-04-01

    The influence of preparation time on ankle joint biomechanics during highly dynamic movements is largely unknown. The aim of this study was to evaluate the impact of limited preparation time on ankle joint loading during highly dynamic run-and-cut movements. Thirteen male basketball players performed 45°-sidestep-cutting and 180°-turning manoeuvres in reaction to light signals which appeared during the approach run. Both movements were executed under (1) an easy condition, in which the light signal appeared very early, (2) a medium condition and (3) a hard condition with very little time to prepare the movements. Maximum ankle inversion angles, moments and velocities during ground contact, as well as EMG signals of three lower extremity muscles, were analysed. In 180°-turning movements, reduced preparation time led to significantly increased maximum ankle inversion velocities. Muscular activation levels, however, did not change. Increased inversion velocities, without accompanying changes in muscular activation, may have the potential to destabilise the ankle joint when less preparation time is available. This may result in a higher injury risk during turning movements and should therefore be considered in ankle injury research and the aetiology of ankle sprains.

  12. Metal glass vacuum tube solar collectors are approaching lower-medium temperature heat application.

    PubMed

    Jiang, Xinian

    2010-04-26

    Solar thermal collectors are widely used worldwide mainly for hot water preparation at a low temperature (less than 80 degrees C). Applications including many industrial processes and central air conditioning with absorption chillers, instead require lower-medium temperature heat (between 90 degrees C and 150 degrees C) to be driven when using solar thermal energy. The metal absorber glass vacuum tube collectors (MGVT) are developed for this type of applications. Current state-of-art and possible future technology development of MGVT are presented.

  13. Physicochemical properties of precursors of Al2O3-ZrO2 oxide ceramics prepared by electrochemical method

    NASA Astrophysics Data System (ADS)

    Petrova, E. V.; Dresvyannikov, A. F.; Ahmadi Daryakenari, M.; Khairullina, A. I.

    2016-05-01

    Scanning electron microscopy, X-ray, and thermal analysis are used to examine the structure and properties of dispersive systems based on aluminum and zirconium oxides prepared electrochemically. The effect the conditions of synthesis have on the structure and morphology of Al2O3-ZrO2 particles is studied. It is shown that the effect of an electric field on the reaction medium allows us to adjust the physicochemical properties and morphology.

  14. Effects of biomaterial-derived fibroblast conditioned medium on the α-amylase expression of parotid gland acinar cells.

    PubMed

    Chou, Ya-Shuan; Young, Tai-Horng; Lou, Pei-Jen

    2015-11-01

    Salivary gland cells are surrounded by a complex stromal environment, in which fibroblasts are the main cells in proximity to the gland cells. In this study, the interaction between parotid gland acinar cells (PGACs), fibroblasts, and biomaterials was investigated. We prepared different biomaterials, including chitosan, polyvinyl alcohol (PVA), poly (ethylene-co-vinyl alcohol) (EVAL), polyvinylidene fluoride (PVDF), and tissue culture polystyrene (TCPS) to culture fibroblasts and then collect their conditioned media to culture PGACs. We observed no difference in AQP3, AQP5, and E-cadherin expression among different fibroblast conditioned medium treatments. Interestingly, α-amylase expression was obviously enhanced in PGACs cultured in the presence of conditioned medium from fibroblasts cultured on PVDF. Higher neurotrophin-4 (NT-4) expression was observed in PVDF-derived fibroblast conditioned medium using a growth factor protein array assay. In addition, directly adding NT-4 into the culture medium significantly promoted α-amylase expression by PGACs. Finally, nestin and βIII-tubulin expression by fibroblasts cultured on PVDF was also enhanced. Together, these results suggest that PVDF could promote α-amylase expression by PGACs via the NT-4 produced by fibroblasts. To date, there is no effective therapy for patients with dry mouth with persistent salivary hypofunction. The study made use of different biomaterials to culture fibroblasts and then collect their conditioned media to culture PGACs. It was found that the effect of fibroblast conditioned medium from PVDF on the α-amylase expression of PGACs was obviously enhanced and higher neurotrophin-4 (NT-4) expression was found in PVDF-derived fibroblast conditioned medium. In addition, directly adding NT-4 into the culture medium significantly promoted the expression of α-amylase by PGACs and the expression of nestin and βIII-tubulin of fibroblasts after being cultured on PVDF was enhanced. Therefore, the present study represents the first description of the role of NT-4 in the expression of α-amylase of PGACs and the role of PVDF in the reprogramming fibroblasts into neural progenitor-like cells, indicating that PVDF could promote the expression of α-amylase by PGACs via the NT-4 produced by fibroblasts. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  15. Cultivation of Arthrospira (spirulina) platensis in desalinator wastewater and salinated synthetic medium: protein content and amino-acid profile

    PubMed Central

    Volkmann, Harriet; Imianovsky, Ulisses; Oliveira, Jorge L.B.; Sant’Anna, Ernani S.

    2008-01-01

    Arthrospira (Spirulina) platensis was cultivated in laboratory under controlled conditions (30°C, photoperiod of 12 hours light/dark provided by fluorescent lamps at a light intensity of 140 μmol photons.m-2.s-1 and constant bubbling air) in three different culture media: (1) Paoletti medium (control), (2) Paoletti supplemented with 1 g.L-1 NaCl (salinated water) and (3) Paoletti medium prepared with desalinator wastewater. The effects of these treatments on growth, protein content and amino acid profile were measured. Maximum cell concentrations observed in Paoletti medium, Paoletti supplemented with salinated water or with desalinator wastewater were 2.587, 3.545 and 4.954 g.L-1, respectively. Biomass in medium 3 presented the highest protein content (56.17%), while biomass in medium 2 presented 48.59% protein. All essential amino acids, except lysine and tryptophan, were found in concentrations higher than those requiried by FAO. PMID:24031187

  16. Inactivation of Escherichia coli and coliform bacteria in traditional brass and earthernware water storage vessels.

    PubMed

    Tandon, Puja; Chhibber, Sanjay; Reed, Robert H

    2005-07-01

    The detection and enumeration of indicator bacteria such as Escherichia coli is used to assess the extent of faecal contamination of drinking water. On the basis of this approach, the effectiveness of storing water contaminated with faecal indicator bacteria in brass or earthern vessels (mutkas) of the type used in rural India have been investigated. Suspensions of bacteria in sterile distilled water were maintained for up to 48 h in each vessel and enumerated by surface plate counts on nutrient agar (non-selective) and several selective coliform media at 37 degrees C either under standard aerobic conditions, or under conditions designed to neutralise reactive oxygen species (ROS), e.g. using an anaerobic cabinet to prepare plates of pre-reduced growth medium or by inclusion of sodium pyruvate in the growth medium, with incubation of aerobically-prepared plates in an anaerobic jar. The counts obtained for E. coli decreased on short-term storage in a brass mutka; counts for selective media were lower than for equivalent counts for non-selective medium, with ROS-neutralised conditions giving consistently higher counts than aerobic incubation. However, after 48 h, no bacteria were cultivable under any conditions. Similar results were obtained using water from environmental sources in the Panjab, and from rural households where brass and earthern mutkas are used for storage of drinking water, with enumeration on selective coliform media (presumptive total coliforms). In all cases results indicated that, while storage of water in a brass mutka can inactivate E. coli and coliforms over a 48 h period, standard aerobic plate counting using selective media may not be fully effective in enumerating sub-lethally damaged bacteria.

  17. Eco-friendly microwave-assisted protocol to prepare hyaluronan-fatty acid conjugates and to induce their self-assembly process.

    PubMed

    Calce, Enrica; Mercurio, Flavia Anna; Leone, Marilisa; Saviano, Michele; De Luca, Stefania

    2016-06-05

    An environmentally sustainable and energy-efficient synthetic process has been developed to prepare hyaluronan-based nano-sized material. It consists in a microwave-promoted acylation of the hydroxyl function of the polysaccharide with natural fatty acids, performed under solvent-free conditions. The efficient interaction of the solid reagents with the MW radiation accounts for the obtained high yielded products. The self-assembly process of the obtained compounds very fast occurred in an aqueous medium under MW-radiation, thus allowing the development of a green protocol for the nano-particles preparation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Microbial fuel cell with improved anode

    DOEpatents

    Borole, Abhijeet P.

    2010-04-13

    The present invention relates to a method for preparing a microbial fuel cell, wherein the method includes: (i) inoculating an anodic liquid medium in contact with an anode of the microbial fuel cell with one or more types of microorganisms capable of functioning by an exoelectrogenic mechanism; (ii) establishing a biofilm of the microorganisms on and/or within the anode along with a substantial absence of planktonic forms of the microorganisms by substantial removal of the planktonic microorganisms during forced flow and recirculation conditions of the anodic liquid medium; and (iii) subjecting the microorganisms of the biofilm to a growth stage by incorporating one or more carbon-containing nutritive compounds in the anodic liquid medium during biofilm formation or after biofilm formation on the anode has been established.

  19. Modelling of the Nutrient Medium for Plants Cultivation in Spaceflight

    NASA Astrophysics Data System (ADS)

    Nechitailo, Galina S.

    2016-07-01

    MODELLING OF THE NUTRIENT MEDIUM FOR PLANTS CULTIVATION IN SPACEFLIGHT Nechitajlo G.S.*, Rakhmetova A.A.**, Bogoslovskaja O.A.**, Ol'hovskay I.P.**, Glushchenko N.N.** *Emanuel Institute of Biochemical Physics of Russian Academy of Sciences (IBCP RAS) mail: spacemal@mail.ru **V.L. Talrose Institute for Energy Problems of Chemical Physics of Russian Academy of Science (INEPCP RAS) mail: nnglu@ mail.ru The valuable life and fruitful activity of cosmonauts and researchers in conditions of spaceflights and prolonged work at space stations are only possible with creating life area providing fresh air, natural food, comfortable psychological conditions, etc. The solution of that problem under space conditions seems impossible without use of high nano- and biotechnologies for plants growth. A priority should be given not only to choose species of growth plants in space, but also to improve conditions for their growth which includes optimal nourishing components for plants, preparation of nutrient mediums, illumination and temperature. We are deeply convinced that just manipulations with growing conditions for cultivated plants, but not genes changes, is a guarantee of success in the decision of this problem. For improving the method of plants growing on the artificial nutrient medium with balanced content of components, being necessary for growth and development of plants, we added essential metal elements: Fe, Zn, Cu - in an electroneutral state in the form of nanoparticles instead of sulfates or other easily dissolving salts. Nanoparticulated metals are known to have a number of advantages in comparison with salts: metals in an electroneutral form are characterized with the prolonged and multifunctional action, low toxicity per se and appearing to be much below the toxicity of the same metals in the ionic forms, accumulation as a reserve being used in biotic dozes, active distribution in bodies and organs of plants and stimulation of vital processes. A high reactivity of nanoparticles and their active interactions with components of a nutrient medium demands development of certain technological solutions for conservation of activity potential of nanoparticles in nutrient mediums. Thus, we have elaborated and created the artificial nutrient medium having balanced structure of components and assuring successful plants cultivation in conditions of spaceflight.

  20. Casting ability of selected impression materials tested in different conditions in an in vitro sulcus model.

    PubMed

    Kolbeck, Carola; Rosentritt, Martin; Lang, Reinhold; Schiller, Manuela; Handel, Gerhard

    2009-10-01

    To test casting capacities of impression materials under dry and wet sulcular conditions in vitro. An incisor with a circular shoulder preparation (1 mm) was inserted in a primary mold. A shiftable secondary mold allowed adaptation of sulcular depth (1 to 4 mm). An outer circular chamfer assured reproducible positioning of an impression material carrier. Tested materials were PVS of differing viscosities (extra low, Panasil Contact Plus [ELV]; low, Affinis Light Body [LV]; and medium, Virtual Monophase [MV]) and one polyether material of low viscosity (Permadyne Garant [PE]). Impressions were made with sulcular depths of 1 to 4 mm in wet and 1 and 4 mm in dry conditions, cut in half, and digitized with a light microscope (Stemi SV8). Surface area of the region of interest (ROI, at inner angle of preparation) was determined with Optimas 6.2. Medians were calculated, and statistical analysis was performed using the Mann-Whitney U test (P # .05). Median values of the measurements under wet condition demonstrated the smallest ROI areas for the ELV (297-330[pixel]) and the MV (253-421[pixel]) materials followed by the LV (582-745[pixel]) and the PELV (544-823[pixel]). All materials showed significantly higher values for the wet compared to dry sulcular conditions. Repeated measurements showed no significant differences to the corresponding first determined series. The sulcus model is applicable to assess casting abilities of impression materials in clinically approximated sulcular conditions. The PVS materials with extra low and medium viscosities showed the best properties in dry and wet conditions.

  1. Development of CMC hydrogels loaded with silver nano-particles for medical applications.

    PubMed

    Hebeish, Ali; Hashem, M; El-Hady, M M Abd; Sharaf, S

    2013-01-30

    Innovative CMC-based hydrogels with great potentials for usage in medical area were principally synthesized as per two strategies .The first involved reaction of epichlorohydrin in alkaline medium containing silver nitrate to yield silver nano-particles (AgNPs)-loaded CMC hydrogel. While CMC acted as stabilizing for AgNPs, trisodium citrate was added to the reaction medium to assist CMC in establishing reduction of Ag(+) to AgNPs. The second strategy entailed preparation of CMC hydrogel which assists the in situ preparation of AgNPs under the same conditions. In both strategies, factors affecting the characterization of AgNPs-loaded CMC hydrogels were studied. Analysis and characterization of the so obtained hydrogels were performed through monitoring swelling behavior, FTIR spectroscopy, SEM, EDX, UV-vis spectrophotometer and TEM. Antimicrobial activity of the hydrogels was examined and mechanisms involved in their synthesis were reported. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. [Study on preparation of sagittatoside B with epimedin B converted from cellulase].

    PubMed

    Xu, Feng-Juan; Sun, E; Zhang, Zhen-Hai; Cui, Li; Jia, Xiao-Bin

    2014-01-01

    To prepare sagittatoside B with epimedin B Hydrolyzed from cellulase. With the conversion ratio as the index, the effects of pH value, temperature, reaction time, dosage of enzyme and concentration of substrates on the conversion ratio were detected. L9 (3(4)) orthogonal design was adopted to optimize the preparation process. Hydrolyzed products were identified by MS, 1H-NMR, and 13C-NMR. The results showed that the optimum reaction conditions for the enzymatic hydrolysis were that the temperature was 50 degrees C, the reaction medium was pH 5.6 acetic acid-sodium acetate buffer solution, the concentration of substrates was 20 g x L(-1), the mass ratio between enzyme and substrate was 3: 5, and the relative molecular mass of the reaction product was 646.23. NMR data proved that the product was sagittatoside B. The process is simple and reliable under mild reaction conditions, thus suitable for industrial production.

  3. Structural, optical and magnetic studies of CuFe2O4, MgFe2O4 and ZnFe2O4 nanoparticles prepared by hydrothermal/solvothermal method

    NASA Astrophysics Data System (ADS)

    Kurian, Jessyamma; Mathew, M. Jacob

    2018-04-01

    In this paper we report the structural, optical and magnetic studies of three spinel ferrites namely CuFe2O4, MgFe2O4 and ZnFe2O4 prepared in an autoclave under the same physical conditions but with two different liquid medium and different surfactant. We use water as the medium and trisodium citrate as the surfactant for one method (Hydrothermal method) and ethylene glycol as the medium and poly ethylene glycol as the surfactant for the second method (solvothermal method). The phase identification and structural characterization are done using XRD and morphological studies are carried out by TEM. Cubical and porous spherical morphologies are obtained for hydrothermal and solvothermal process respectively without any impurity phase. The optical studies are carried out using FTIR and UV-Vis reflectance spectra. In order to elucidate the nonlinear optical behaviour of the prepared nanomaterial, open aperture z-scan technique is used. From the fitted z-scan curves nonlinear absorption coefficient and the saturation intensity are determined. The magnetic characterization of the samples is performed at room temperature using vibrating sample magnetometer measurements. The M-H curves obtained are fitted using theoretical equation and the different components of magnetization are determined. Nanoparticles with high saturation magnetization are obtained for MgFe2O4 and ZnFe2O4 prepared under solvothermal reaction. The magnetic hyperfine parameters and the cation distribution of the prepared materials are determined using room temperature Mössbauer spectroscopy. The fitted spectra reveal the difference in the magnetic hyperfine parameters owing to the change in size and morphology.

  4. Preparation and characterization of V/TiO{sub 2} nanocatalyst with magnetic nucleus of iron

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

    Feyzi, Mostafa; Rafiee, Hamid Reza, E-mail: rafieehr@yahoo.com; Ranjbar, Shahram

    2013-11-15

    Graphical abstract: - Highlights: • Fe-V/TiO{sub 2} nanocatalyst is prepared. • Combination of sol–gel and wetness impregnation methods. • Facile separation of catalyst from medium by magnet. - Abstract: A magnetic composite containing V/TiO{sub 2} was prepared by combination of sol–gel and wetness impregnation methods. The effects of synthesis temperature, different weight percents of Fe supported on TiO{sub 2}, vanadium loading and the heating rate of calcination on the structure and morphology of nanocatalyst were investigated. The optimum conditions for synthesized catalyst were 40 wt.% of Fe, 15 wt.% of V and synthesis temperature equal to 30 °C. Characterization ofmore » catalyst is carried out using XRD, TGA, DSC, SEM, FTIR and N{sub 2} physisorption measurements. The magnetic character of nanocatalyst was measured using VSM, which showed the typical paramagnetic behavior of sample at room temperature with a saturation magnetization value equal to 8.283 emu/g. The nanocatalyst has a particle size about 56 nm and can easily be separated from medium by a magnet.« less

  5. Testicular cell conditioned medium supports differentiation of embryonic stem cells into ovarian structures containing oocytes.

    PubMed

    Lacham-Kaplan, Orly; Chy, Hun; Trounson, Alan

    2006-02-01

    Previous reports and the current study have found that germ cell precursor cells appear in embryoid bodies (EBs) formed from mouse embryonic stem cells as identified by positive expression of specific germ cell markers such as Oct-3/4, Mvh, c-kit, Stella, and DAZL. We hypothesized that if exposed to appropriate growth factors, the germ cell precursor cells within the EBs would differentiate into gametes. The source for growth factors used in the present study is conditioned medium collected from testicular cell cultures prepared from the testes of newborn males. Testes at this stage of development contain most growth factors required for the transformation of germ stem cells into differentiated gametes. When EBs were cultured in the conditioned medium, they developed into ovarian structures, which contained putative oocytes. The oocytes were surrounded by one to two layers of flattened cells and did not have a visible zona pellucida. However, oocyte-specific markers such as Fig-alpha and ZP3 were found expressed by the ovarian structures. The production of oocytes using this method is repeatable and reliable and may be applicable to other mammalian species, including the human.

  6. Synthesis, characterization and photocatalytic activity of porous manganese oxide doped titania for toluene decomposition.

    PubMed

    Jothiramalingam, R; Wang, M K

    2007-08-17

    The present study describes the photocatalytic degradation of toluene in gas phase on different porous manganese oxide doped titanium dioxide. As synthesized birnessite and cryptomelane type porous manganese oxide were doped with titania and tested for photocatalytic decomposition of toluene in gas phase. The effects of the inlet concentration of toluene, flow rate (retention time) were examined and the relative humidity was maintained constantly. Thermal and textural characterization of manganese oxide doped titania materials were characterized by X-ray diffraction (XRD), thermogravemetry (TG), BET and TEM-EDAX studies. The aim of the present study is to synthesize the porous manganese oxide doped titania and to study its photocatalytic activity for toluene degradation in gas phase. Cryptomelane doped titania catalyst prepared in water medium [K-OMS-2 (W)] is shown the good toluene degradation with lower catalysts loading compared to commercial bulk titania in annular type photo reactor. The higher photocatalytic activity due to various factors such as catalyst preparation method, experimental conditions, catalyst loading, surface area, etc. In the present study manganese oxide OMS doped titania materials prepared by both aqueous and non-aqueous medium, aqueous medium prepared catalyst shows the good efficiency due to the presence of OH bonded groups on the surface of catalyst. The linear forms of different kinetic equations were applied to the adsorption data and their goodness of fit was evaluated based on the R2 and standard error. The goodness to the linear fit was observed for Elovich model with high R2 (>or=0.9477) value.

  7. An investigation on the preparation of nanocrystalline hydrous zirconia from zirconium tungstate

    NASA Astrophysics Data System (ADS)

    Antunes, M.; Perottoni, C. A.; Gouvêa, D.; Machado, G.; Zorzi, J. E.

    2018-02-01

    Hydrous nanocrystalline zirconia was prepared from an unusual precursor—the bimetallic oxide zirconium tungstate (ZrW2O8)—in alkaline medium. Different experimental conditions (NaOH concentration, time and temperature) were used to investigate the effects on crystallographic, morphological, chemical and thermal characteristics of the products. The resulting materials are composed of particles with a crystal structure similar to that of cubic ZrO2 (or a mixture of tetragonal and cubic phases, depending on the synthesis conditions), with particle size around 5 nm and crystallites around 3 nm in diameter. These particles form high surface area agglomerates, exhibiting mesoporosity and capacity for adsorption of water and carbon dioxide. The synthesis mechanism appears to be constituted, first, by a chemical substitution reaction between the WO4 tetrahedra and hydroxyl ions, with subsequent solubilization of the structure. Indeed, excess hydroxyls in the medium form colloidal zirconium ions which polymerize/condense, generating crystalline nuclei in a process facilitated by heterogeneous nucleation and supersaturation. The presence of residual tungsten in all samples appears to be a key element for stabilizing the size and crystalline structure of the materials produced.

  8. The role of medium radiation dose on microbiological safety and shelf-life of some traditional soups

    NASA Astrophysics Data System (ADS)

    Irawati, Z.; Natalia, L.; Nurcahya, C. M.; Anas, F.

    2007-11-01

    Irradiation at medium doses in combination with cryogenic condition along the process to ensure the safety, quality and to extend the shelf-life of prepared meals have been investigated. Semi-concentrated black, ox-tail, chicken vegetable and chicken sweet corn soups were individually packed in a dry laminate pouch of PET 12 μ/LDPE adh.2 μ/Al-foil 7 μ/LDPE adh/LLDPE (C 4) 50 μ under vacuum followed by freezing for 24 h at -18 °C prior to irradiation with doses of 1, 3, 5 and 7 kGy at cryogenic condition (-79 °C), respectively. Both the non-irradiated and irradiated prepared meals were then stored in refrigerator at 5±2 °C. Non-irradiated and the irradiated samples at 1 kGy were mostly damaged after a week of storage. Gamma irradiation at doses of 5-7 kGy for the soups could reduce microbial load by about 2-3 log cycles, respectively, without affecting the physical-chemical parameters and palatability within 2-3 months while the unirradiated samples could only withstand for 1 month storage time.

  9. A novel method to prepare L-Arabinose from xylose mother liquor by yeast-mediated biopurification

    PubMed Central

    2011-01-01

    Background L-arabinose is an important intermediate for anti-virus drug synthesis and has also been used in food additives for diets-controlling in recent years. Commercial production of L-arabinose is a complex progress consisting of acid hydrolysis of gum arabic, followed by multiple procedures of purification, thus making high production cost. Therefore, there is a biotechnological and commercial interest in the development of new cost-effective and high-performance methods for obtaining high purity grade L-arabinose. Results An alternative, economical method for purifying L-arabinose from xylose mother liquor was developed in this study. After screening 306 yeast strains, a strain of Pichia anomala Y161 was selected as it could effectively metabolize other sugars but not L-arabinose. Fermentation in a medium containing xylose mother liquor permitted enrichment of L-arabinose by a significant depletion of other sugars. Biochemical analysis of this yeast strain confirmed that its poor capacity for utilizing L-arabinose was due to low activities of the enzymes required for the metabolism of this sugar. Response surface methodology was employed for optimization the fermentation conditions in shake flask cultures. The optimum conditions were: 75 h fermentation time, at 32.5°C, in a medium containing 21% (v/v) xylose mother liquor. Under these conditions, the highest purity of L-arabinose reached was 86.1% of total sugar, facilitating recovery of white crystalline L-arabinose from the fermentation medium by simple methods. Conclusion Yeast-mediated biopurification provides a dynamic method to prepare high purity of L-arabinose from the feedstock solution xylose mother liqour, with cost-effective and high-performance properties. PMID:21649890

  10. Influence of coating defects on the corrosion behavior of cold sprayed refractory metals

    NASA Astrophysics Data System (ADS)

    Kumar, S.; Rao, A. Arjuna

    2017-02-01

    The defects in the cold sprayed coatings are critical in the case of corrosion performances of the coatings in aggressive conditions. To understand the influence of coating defects on corrosion, immersion tests have been carried out in HF solution for the cold sprayed and heat treated Titanium, Tantalum and Niobium coatings. Long duration immersion tests reveal inhomogeneous weight losses of the samples prepared at different heat treatment conditions. The weight loss for different coatings has been well corroborated with the coating defects and microstructures. Chemical and micro structural analysis elucidates the reason behind the inhomogeneous performance of different type of cold sprayed coatings in corrosion medium. In the case of cold sprayed titanium, formation of stable oxide along the inter-splat boundary hinders the aggressive attack of the corrosion medium which is not so in other cases.

  11. Preparation of anti-inflammatory mesenchymal stem/precursor cells (MSCs) through sphere formation using hanging-drop culture technique.

    PubMed

    Bartosh, Thomas J; Ylostalo, Joni H

    2014-02-06

    Herein, we describe a protocol for preparation of pre-activated anti-inflammatory human mesenchymal stem/precursor cells (MSCs) in 3-D culture without addition of exogenous chemicals or gene-transfer approaches. MSCs are an easily procurable source of multipotent adult stem cells with therapeutic potential largely attributed to their paracrine regulation of inflammation and immunity. However, the culture conditions to prepare the ideal MSCs for cell therapy remain elusive. Furthermore, the reported lag time for activation in experimental models has prompted investigations on pre-activating the cells prior to their administration. In this protocol, standard 2-D culture-expanded MSCs are activated by aggregation into 3-D spheres using hanging-drop cultures. MSC activation is evaluated by real-time PCR and/or ELISA for anti-inflammatory factors (TSG-6, STC-1, PGE2), and by a functional assay using lipopolysaccharide-stimulated macrophage cultures. Further, we elucidate methods to prepare MSC-sphere conditioned medium, intact spheres, and suspension of single cells from spheres for experimental and clinical applications. Copyright © 2014 John Wiley & Sons, Inc.

  12. Preparation of anti-inflammatory mesenchymal stem/precursor cells (MSCs) through sphere formation using hanging drop culture technique

    PubMed Central

    Bartosh, Thomas J.

    2014-01-01

    Herein, we describe a protocol for preparation of pre-activated anti-inflammatory human mesenchymal stem/precursor cells (MSCs) in 3D culture without addition of exogenous chemicals or gene transfer approaches. MSCs are an easily procurable source of multipotent adult stem cells with therapeutic potential largely attributed to their paracrine regulation of inflammation and immunity. However, the culture conditions to prepare the ideal MSCs for cell therapy remain elusive. Furthermore, reported lag time for activation in experimental models have prompted investigations to pre-activate the cells prior to their administration. In this protocol, standard 2D culture expanded MSCs are activated by aggregation into 3D spheres using hanging drop cultures. MSC activation is evaluated by real-time PCR and/or ELISA for anti-inflammatory factors (TSG-6, STC-1, PGE2), and by a functional assay using lipopolysaccharide-stimulated macrophage cultures. Furthermore, we elucidate methods to prepare MSC sphere conditioned medium, intact spheres, and suspension of single cells from spheres for experimental and clinical applications. PMID:24510769

  13. Polymer-based adsorption medium prepared using a fragment imprinting technique for homologues of chlorinated bisphenol A produced in the environment.

    PubMed

    Kubo, Takuya; Hosoya, Ken; Watabe, Yoshiyuki; Ikegami, Tohru; Tanaka, Nobuo; Sano, Tomoharu; Kaya, Kunimitsu

    2004-03-12

    A polymer-based adsorption medium having molecular recognition ability for homologues of chlorinated bisphenol A produced in environment was prepared using a fragment imprinting technique. 2,6-Dimethyl phenol was utilized as a pseudo-template molecule and the adsorption media prepared was evaluated by high performance liquid chromatography (HPLC) and solid-phase extraction (SPE). As results, the adsorption medium showed preferable chromatographic retention and specific adsorption ability for the chlorinated bisphenol As having chlorine substituents at 3,5-positions through fragment imprinting effect.

  14. New method for the isolation of Streptococcus mutans and its differentiation from other oral streptococci.

    PubMed

    Linke, H A

    1977-06-01

    A new, improved agar medium for the isolation of Streptococcus mutans, the etiological agent of dental caries, was developed. In contrast to mitis-salivarius agar, this medium not only recovers a greater number of S. mutans strains from most oral specimens but, because of its mannitol and sorbitol content, it also facilitates the differentiation of S. mutans from other oral streptococci, e.g., S. salivarius, S. mitis, and S. sanguis, which do not grow or produce scanty growth only after 10 days of incubation. The medium is easy to prepare because of its simple and unique composition, is characterized by the presence of an acid indicator, and can be utilized under aerobic and anaerobic conditions as well. The medium cannot be used to distinguish among the eight serotypes, a to g and SL-1, of S. mutans. Mannitol-utilizing bacteria such as streptococci (e.g., S. faecalis) and other microorganisms (e.g., Staphylococcus aureus) are able to grow on this medium and can be distinguished from S. mutans by their unique colony morphology.

  15. Control of 5-aminolaevulinate synthetase activity in Rhodopseudomonas spheroides. The involvement of sulphur metabolism

    PubMed Central

    Neuberger, Albert; Sandy, John D.; Tait, George H.

    1973-01-01

    1. The `initial' 5-aminolaevulinate synthetase activity, that is the activity observed immediately after cell disruption, in extracts prepared from unharvested semianaerobically grown Rhodopseudomonas spheroides, was twice that observed under the same assay conditions in extracts prepared from harvested cells. 2. The effect of oxygenation of a culture on the `maximum' aminolaevulinate synthetase activity, that is the activity observed 1h after disruption of harvested cells, is markedly influenced by the contents of the growth medium. Oxygenation of organisms for 1h in the medium in which they have grown produces an 80–90% decrease in maximum activity, whereas similar treatment of organisms resuspended in fresh medium produces less than a 40% decrease. 3. This protective effect of fresh medium is absolutely dependent on the presence of sulphate. When cells are suspended in sulphate-deficient fresh medium, the maximum activity falls by 65–75% even without oxygenation. A high maximum activity is regenerated when sulphate is resupplied. 4. When organisms are oxygenated in the medium in which they have grown, the cellular contents of GSH+GSSG and cysteine+cystine fall very markedly and homolanthionine is formed. Both the fall in aminolaevulinate synthetase activity and the changes in sulphur metabolism are largely prevented by the addition of compounds which stimulate synthesis of cysteine de novo or inhibit the conversion of cysteine S into homocysteine S. 5. The maximum aminolaevulinate synthetase activity was directly proportional to the GSH+GSSG content of all cell preparations. In glutathione-depleted extracts the `low'-activity enzyme could be re-activated in vitro by the addition of GSH, GSSG, cysteine or cystine, whereas in extracts with a high glutathione content the `high'-activity enzyme was unaffected by these sulphur compounds. 6. The activation of low-activity enzyme with exogenous sulphur compounds was prevented by excluding air or by adding NADH. Studies with purified enzyme indicate that sulphur compounds do not interact directly with the enzyme, but that their effect is mediated by a number of other endogenous factors. PMID:4544404

  16. A novel sample preparation method to avoid influence of embedding medium during nano-indentation

    Treesearch

    Yujie Meng; Siqun Wang; Zhiyong Cai; Timothy M. Young; Guanben Du; Yanjun Li

    2012-01-01

    The effect of the embedding medium on the nano-indentation measurements of lignocellulosic materials was investigated experimentally using nano-indentation. Both the reduced elastic modulus and the hardness of nonembedded cell walls were found to be lower than those of the embedded samples, proving that the embedding medium used for specimen preparation on cellulosic...

  17. Silica adsorbent prepared from spent diatomaceous earth and its application to removal of dye from aqueous solution.

    PubMed

    Tsai, W T; Hsien, K J; Yang, J M

    2004-07-15

    The objective of this work is to study the activation regeneration of spent diatomaceous earth (SDE) for the preparation of silica adsorbents using thermal regeneration and acid/alkaline activation methods. Under the experimental conditions investigated, it was found that the alkaline activation method carried out by sodium hydroxide under controlled conditions is significantly superior to other heat and activation methods. The porosities of solids thus obtained are over 0.2, indicating that they are basically mesoporous. The optimal porous material thus prepared was used as a mineral adsorbent for methylene blue at 25 degrees C. The adsorption equilibrium revealed that the silica adsorbent can take up over 50 mg/g at relatively low concentrations in aqueous medium from the fittings of Langmuir and Freundlich isotherms with high correlations. On the other hand, the adsorption kinetic of methylene blue under various adsorbent dosages can be well described with a pseudo-second-order reaction model. Copyright 2004 Elsevier Inc.

  18. Physico-chemical characteristics of burfi prepared by using medium chain triglyceride rich margarines.

    PubMed

    Tiwari, Shipra; Chetana, Ramakrishna; Puttaraju, Shashikala; Khatoon, Sakina

    2014-01-01

    Medium chain triglyceride rich margarines were prepared using palm, coconut oil blends in the ratio of 80:20 (Margarine 1) and 60:40 (Margarine 2). The margarines were used to prepare burfi and compared with products prepared using commercial margarine, ghee and butter. The physicochemical characteristics such as texture, color, free fatty acid, peroxide value, saponification value, unsaponifiable matter and fatty acid composition of oils, fats and margarines were carried out. Results showed that 11.0 and 21.9% of medium chain triglycerides were present in margarine 1 and 2 respectively. The texture, colour, moisture content, peroxide value and sensory evaluation were carried out for the burfi samples. Laboratory prepared margarines improved the textural quality of burfi compared to commercial margarine, ghee and butter. The sensory analyses of the burfi samples revealed that burfi prepared from margarine 1 was more acceptable compared to commercial margarine.

  19. Oxygen reduction reaction catalyzed by platinum nanonetwork prepared by template free one step synthesis for polymer electrolyte membrane fuel cells

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

    Narayanamoorthy, B.; Kumar, B.V.V.S. Pavan; Eswaramoorthy, M.

    2014-07-01

    Highlights: • Supportless Pt nanonetwork (Pt NN) synthesized by novel template free one step method as per our earlier reported procedure. • Electrocatalytic activity of Pt NN studied taking oxygen reduction reaction in acid medium. • Kinetic and thermodynamic parameters were deduced under hydrodynamic conditions. • ORR mechanistic pathway was proposed based on kinetic rate constants. • ADT analysis found enhanced stability (5000 cycles) for Pt NN than Pt NN/VC and reported Pt/C. - Abstract: The reduction reaction of molecular oxygen (ORR) was investigated using supportless Pt nanonetwork (Pt NN) electrocatalyst in sulfuric acid medium. Pt NN was prepared bymore » template free borohydride reduction. The transmission electron microscope images revealed a network like nano-architecture having an average cluster size of 30 nm. The electrochemical characterization of supportless and Vulcan carbon supported Pt NN (Pt NN/VC) was carried out using rotating disc and ring disc electrodes at various temperatures. Kinetic and thermodynamic parameters were estimated under hydrodynamic conditions and compared with Pt NN/VC and reported Pt/C catalysts. The accelerated durability test revealed that supportless Pt NN is quite stable for 5000 potential cycles with 22% reduction in electrochemical surface area (ECSA). While the initial limiting current density has in fact increased by 11.6%, whereas Pt NN/VC suffered nearly 55% loss in ECSA and 13% loss in limiting current density confirming an enhanced stability of supportless Pt NN morphology for ORR compared to conventional Pt/C ORR catalysts in acid medium.« less

  20. National Aerospace Leadership Initiative - Phase I

    DTIC Science & Technology

    2008-09-30

    Devised and validated CFD code for operation of a micro-channel heat exchanger. The work was published at the 2008 AIAA Annual Meeting and Exposition...and (3) preparation to implement this algorithm in TURBO. Heat Transfer Capability In the short and medium term, the following plan has been adopted...to provide heat transfer capability to the TURBO code: • Incorporation of a constant wall temperature boundary condition. This capability will be

  1. Medium-Chain Fatty Acids Improve Cognitive Function in Intensively Treated Type 1 Diabetic Patients and Support In Vitro Synaptic Transmission During Acute Hypoglycemia

    PubMed Central

    Page, Kathleen A.; Williamson, Anne; Yu, Namyi; McNay, Ewan C.; Dzuira, James; McCrimmon, Rory J.; Sherwin, Robert S.

    2009-01-01

    OBJECTIVE We examined whether ingestion of medium-chain triglycerides could improve cognition during hypoglycemia in subjects with intensively treated type 1 diabetes and assessed potential underlying mechanisms by testing the effect of β-hydroxybutyrate and octanoate on rat hippocampal synaptic transmission during exposure to low glucose. RESEARCH DESIGN AND METHODS A total of 11 intensively treated type 1 diabetic subjects participated in stepped hyperinsulinemic- (2 mU · kg−1 · min−1) euglycemic- (glucose ∼5.5 mmol/l) hypoglycemic (glucose ∼2.8 mmol/l) clamp studies. During two separate sessions, they randomly received either medium-chain triglycerides or placebo drinks and performed a battery of cognitive tests. In vitro rat hippocampal slice preparations were used to assess the ability of β-hydroxybutyrate and octanoate to support neuronal activity when glucose levels are reduced. RESULTS Hypoglycemia impaired cognitive performance in tests of verbal memory, digit symbol coding, digit span backwards, and map searching. Ingestion of medium-chain triglycerides reversed these effects. Medium-chain triglycerides also produced higher free fatty acids and β-hydroxybutyrate levels compared with placebo. However, the increase in catecholamines and symptoms during hypoglycemia was not altered. In hippocampal slices β-hydroxybutyrate supported synaptic transmission under low-glucose conditions, whereas octanoate could not. Nevertheless, octanoate improved the rate of recovery of synaptic function upon restoration of control glucose concentrations. CONCLUSIONS Medium-chain triglyceride ingestion improves cognition without adversely affecting adrenergic or symptomatic responses to hypoglycemia in intensively treated type 1 diabetic subjects. Medium-chain triglycerides offer the therapeutic advantage of preserving brain function under hypoglycemic conditions without causing deleterious hyperglycemia. PMID:19223595

  2. Quantitative Characterization of the Growth of Deinococcus geothermalis DSM-11302: Effect of Inoculum Size, Growth Medium and Culture Conditions.

    PubMed

    Bornot, Julie; Molina-Jouve, Carole; Uribelarrea, Jean-Louis; Gorret, Nathalie

    2015-08-20

    Due to their remarkable resistance to extreme conditions, Deinococcaceae strains are of great interest to biotechnological prospects. However, the physiology of the extremophile strain Deinococcus geothermalis has scarcely been studied and is not well understood. The physiological behaviour was then studied in well-controlled conditions in flask and bioreactor cultures. The growth of D. geothermalis type strains was compared. Among the strains tested, the strain from the German Collection of Microorganisms (Deutsche Sammlung von Mikroorganismen DSM) DSM-11302 was found to give the highest biomass concentration and growth rate: in a complex medium with glucose, the growth rate reached 0.75 h(-1) at 45 °C. Yeast extract concentration in the medium had significant constitutive and catalytic effects. Furthermore, the results showed that the physiological descriptors were not affected by the inoculum preparation steps. A batch culture of D. geothermalis DSM-11302 on defined medium was carried out: cells grew exponentially with a maximal growth rate of 0.28 h(-1) and D. geothermalis DSM-11302 biomass reached 1.4 g·L(-1) in 20 h. Then, 1.4 gDryCellWeight of biomass (X) was obtained from 5.6 g glucose (Glc) consumed as carbon source, corresponding to a yield of 0.3 CmolX·CmolGlc(-1); cell specific oxygen uptake and carbon dioxide production rates reached 216 and 226 mmol.CmolX(-1)·h(-1), respectively, and the respiratory quotient (QR) value varied from 1.1 to 1.7. This is the first time that kinetic parameters and yields are reported for D. geothermalis DSM-11302 grown on a mineral medium in well-controlled batch culture.

  3. Quantitative Characterization of the Growth of Deinococcus geothermalis DSM-11302: Effect of Inoculum Size, Growth Medium and Culture Conditions

    PubMed Central

    Bornot, Julie; Molina-Jouve, Carole; Uribelarrea, Jean-Louis; Gorret, Nathalie

    2015-01-01

    Due to their remarkable resistance to extreme conditions, Deinococcaceae strains are of great interest to biotechnological prospects. However, the physiology of the extremophile strain Deinococcus geothermalis has scarcely been studied and is not well understood. The physiological behaviour was then studied in well-controlled conditions in flask and bioreactor cultures. The growth of D. geothermalis type strains was compared. Among the strains tested, the strain from the German Collection of Microorganisms (Deutsche Sammlung von Mikroorganismen DSM) DSM-11302 was found to give the highest biomass concentration and growth rate: in a complex medium with glucose, the growth rate reached 0.75 h−1 at 45 °C. Yeast extract concentration in the medium had significant constitutive and catalytic effects. Furthermore, the results showed that the physiological descriptors were not affected by the inoculum preparation steps. A batch culture of D. geothermalis DSM-11302 on defined medium was carried out: cells grew exponentially with a maximal growth rate of 0.28 h−1 and D. geothermalis DSM-11302 biomass reached 1.4 g·L−1 in 20 h. Then, 1.4 gDryCellWeight of biomass (X) was obtained from 5.6 g glucose (Glc) consumed as carbon source, corresponding to a yield of 0.3 CmolX·CmolGlc−1; cell specific oxygen uptake and carbon dioxide production rates reached 216 and 226 mmol.CmolX−1·h−1, respectively, and the respiratory quotient (QR) value varied from 1.1 to 1.7. This is the first time that kinetic parameters and yields are reported for D. geothermalis DSM-11302 grown on a mineral medium in well-controlled batch culture. PMID:27682099

  4. Novel technology to prepare oral formulations for preclinical safety studies.

    PubMed

    Niwa, Toshiyuki; Hashimoto, Naofumi

    2008-02-28

    A novel method to prepare oral formulations, normally suspended dosage form, for preclinical safety studies in animals has been developed using a rotation/revolution mixer. Small hard balls made of zirconia were added to the mixing process to evaluate effectiveness in making a high quality suspension. The driving with balls loaded in the cylindrical container (vessel) of the mixer was quite efficient in dispersing and milling the particles of the active pharmaceutical ingredient (API) in an aqueous medium. The API powder and a small amount of oral aqueous medium (vehicle) were successfully mixed by the spinning motion of the balls in the vessel as though the paste-like suspension was kneaded with a mortar and pestle. It was found that the milled suspension with the mean size of 10-20microm could be prepared, in addition finer milling of less than 10microm could be achieved by selecting the material of vessel. Optimum driving conditions including mixing time, size and quantity of balls, and the standard operational procedure was established using compounds varying in physicochemical properties. The particle size and quantitative analysis by HPLC showed that the resultant suspension was well-milled and highly homogeneous with the nearly intended concentration of API. The proposed method established by this experiment could be applied to the actual safety studies in the real preparation scale of oral suspension.

  5. Method to prepare nanoparticles on porous mediums

    DOEpatents

    Vieth, Gabriel M [Knoxville, TN; Dudney, Nancy J [Oak Ridge, TN; Dai, Sheng [Knoxville, TN

    2010-08-10

    A method to prepare porous medium decorated with nanoparticles involves contacting a suspension of nanoparticles in an ionic liquid with a porous medium such that the particles diffuse into the pores of the medium followed by heating the resulting composition to a temperature equal to or greater than the thermal decomposition temperature of the ionic liquid resulting in the removal of the liquid portion of the suspension. The nanoparticles can be a metal, an alloy, or a metal compound. The resulting compositions can be used as catalysts, sensors, or separators.

  6. Supercritical transitiometry of polymers.

    PubMed

    Randzio, S L; Grolier, J P

    1998-06-01

    Employing supercritical fluids (SCFs) during polymers processing allows the unusual properties of SCFs to be exploited for making polymer products that cannot be obtained by other means. A new supercritical transitiometer has been constructed to permit study of the interactions of SCFs with polymers during processing under well-defined conditions of temperature and pressure. The supercritical transitiometer allows pressure to be exerted by either a supercritical fluid or a neutral medium and enables simultaneous determination of four basic parameters of a transition, i.e., p, T, Δ(tr)H and Δ(tr)V. This permits determination of the SCF effect on modification of the polymer structure at a given pressure and temperature and defines conditions to allow reproducible preparation of new polymer structures. Study of a semicrystalline polyethylene by this method has defined conditions for preparation of new microfoamed phases with good mechanical properties. The low densities and microporous structures of the new materials may make them useful for applications in medicine, pharmacy, or the food industry, for example.

  7. Combination of nutrients in a mammalian cell culture medium kills cryptococci.

    PubMed

    Granger, Donald L; Call, Donna M

    2018-06-06

    We found that a large inoculum of Cryptococcus gattii cells, when plated on Dulbecco's modified eagle's medium (DMEM) incorporated into agar, died within a few hours provided that DMEM agar plates had been stored in darkness for approximately 3 days after preparation. Standard conditions were developed for quantification of killing. The medium lost its fungicidal activity when exposed to visible light of wave length ∼400 nm. The amount of energy required was estimated at 5.8 × 104 joules @ 550 nm. Liquid DMEM conditioned by incubation over DMEM agar plates stored in darkness was fungicidal. We found that fungicidal activity was heat-stable (100°C). Dialysis tubing with MWC0 < 100 Daltons retained fungicidal activity. Neutral pH was required. Strains of Cryptococcus were uniformly sensitive, but some Candida species were resistant. Components of DMEM required for killing were pyridoxal and cystine. Micromolar amounts of iron shortened the time required for DMEM agar plates to become fungicidal when stored in the dark. Organic and inorganic compounds bearing reduced sulfur atoms at millimolar concentrations inhibited fungicidal activity. Our results point to a light-sensitive antifungal compound formed by reaction of pyridoxal with cystine possibly by Schiff base formation.

  8. The quality of sperm preparation medium affects the motility, viability, and DNA integrity of human spermatozoa.

    PubMed

    Anbari, Fatemeh; Halvaei, Iman; Nabi, Ali; Ghazali, Shahin; Khalili, Mohammad Ali; Johansson, Lars

    2016-01-01

    The goal was to compare the effects of three different sperm preparation media on sperm motility, viability, and DNA integrity of semen samples from normozoospermic men. A total of 15 normozoospermic males were included in the study. The semen analysis (SA) was performed in accordance with the WHO guidelines (2010). After SA, each sample was divided into three aliquots, and swim-up was performed with three different sperm preparation media (Sperm Preparation Media, Origio, Denmark; Ham's F10, Biochrome, Berlin, Germany; and VitaSperm™, Innovative Biotech, Iran). Sperm motility, viability, and DNA fragmentation were evaluated at 0, 1, 2, and 24 h after swim-up. There were no significant differences, at any time intervals, in the total sperm motility between the different sperm preparation media. However, the rate of progressive motility was significantly higher in spermatozoa prepared using the media from Origio in comparison with VitaSperm™ ( P = 0.03), whereas no significant difference was found against Ham's F10 medium. No significant differences in sperm viability were seen between the media products. However, 1 h after swim-up, the extent of sperm DNA fragmentation was lower in the medium from Origio versus VitaSperm™ ( P = 0.02). The data showed that the quality of medium for preparation of semen samples from normozoospermic men significantly affects the performance of spermatozoa in assisted conception programs.

  9. Establishment of an in vitro micropropagation protocol for Boscia senegalensis (Pers.) Lam. ex Poir.

    PubMed Central

    Khalafalla, Mutasim M.; Daffalla, Hussien M.; Abdellatef, Eltayb; Agabna, Elsadig; El-Shemy, Hany A.

    2011-01-01

    This report describes in vitro micropropagation of Boscia senegalensis, so-called famine foods, that helped the people in Darfur and Kordofan, Sudan survive during the 1984–1985 famine. Four types of explants prepared from green mature zygotic embryos were cultured on Murashige and Skoog (MS) medium augmented with 1–5 mg/L 6-benzyladenine (BA). The highest number of shoots per explant (14.3±0.9) was achieved on MS medium supplemented with 3 mg/L BA, while the highest shoot length [(3.5±0.4) cm] was obtained with 1 mg/L BA. The shoot cluster, when subcultured to its same medium, significantly increased the rate of shoot multiplication by the end of the third subculture. The maximum mean number of shoots per explant (86.5±3.6) was produced after three multiplication cycles on 3 mg/L BA-supplemented medium. In vitro induced shoots were excised and rooted on half strength MS medium fortified with 0.25 mg/L indole-3-butyric acid (IBA) to obtain complete plantlets. B. senegalensis-regenerated plantlets obtained in vitro for the first time, were hardened and 95% survived under greenhouse conditions. PMID:21462387

  10. Establishment of an in vitro micropropagation protocol for Boscia senegalensis (Pers.) Lam. ex Poir.

    PubMed

    Khalafalla, Mutasim M; Daffalla, Hussien M; Abdellatef, Eltayb; Agabna, Elsadig; El-Shemy, Hany A

    2011-04-01

    This report describes in vitro micropropagation of Boscia senegalensis, so-called famine foods, that helped the people in Darfur and Kordofan, Sudan survive during the 1984-1985 famine. Four types of explants prepared from green mature zygotic embryos were cultured on Murashige and Skoog (MS) medium augmented with 1-5 mg/L 6-benzyladenine (BA). The highest number of shoots per explant (14.3±0.9) was achieved on MS medium supplemented with 3 mg/L BA, while the highest shoot length [(3.5±0.4) cm] was obtained with 1 mg/L BA. The shoot cluster, when subcultured to its same medium, significantly increased the rate of shoot multiplication by the end of the third subculture. The maximum mean number of shoots per explant (86.5±3.6) was produced after three multiplication cycles on 3 mg/L BA-supplemented medium. In vitro induced shoots were excised and rooted on half strength MS medium fortified with 0.25 mg/L indole-3-butyric acid (IBA) to obtain complete plantlets. B. senegalensis-regenerated plantlets obtained in vitro for the first time, were hardened and 95% survived under greenhouse conditions.

  11. Morphology Tuning of Strontium Tungstate Nanoparticles

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

    Joseph, S.; George, T.; George, K. C.

    2007-08-22

    Strontium tungstate nanocrystals in two different morphologies are successfully synthesized by controlled precipitation in aqueous and in poly vinyl alcohol (PVA) medium. Structural characterizations are carried out by XRD and SEM. The average particle size calculated for the SrWO4 prepared in the two different solvents ranges 20-24 nm. The SEM pictures show that the surface morphologies of the SrWO4 nanoparticles in aqueous medium resemble mushroom and the SrWO4 nanoparticles in PVA medium resemble cauliflower. Investigations on the room temperature luminescent properties of the strontium tungstate nanoparticles prepared in aqueous and PVA medium shows strong emissions around 425 nm.

  12. Nanoparticles generated by laser in liquids as contrast medium and radiotherapy intensifiers

    NASA Astrophysics Data System (ADS)

    Restuccia, Nancy; Torrisi, Lorenzo

    2018-01-01

    The synthesis of Au and Ag nanoparticles (NP) though laser ablation in liquids as a function the laser parameters is presented. Spherical NPs with diameter distribution within 1 and 100 nm were prepared by laser ablation in water. The nanoparticles characterization was performed using optical spectroscopy and electronic microscopy (SEM and TEM) measurements. Studies of the possible use of metallic nanoparticles as intensifier of diagnostics imaging contrast medium and absorbing dose from ionizing radiations in traditional radiotherapy and protontherapy are presented. Examples of in vitro (in tissue equivalent materials) and in vivo (in mice), were conducted thank to simulation programs permitting to evaluate the enhancement of efficiency in imaging and therapy as a function of the NPs concentrations and irradiation conditions.

  13. Supercritical fluid technology for enhanced drug delivery and heterogeneous catalysis

    NASA Astrophysics Data System (ADS)

    Pathak, Pankaj

    In recent years there has been great interest in the properties of supercritical fluids (SCFs) and applications of supercritical fluid technology in chemistry, material sciences and many other areas. In our laboratory, we have applied supercritical fluid namely CO2, as a medium for pharmaceutical processing and also for heterogeneous catalysis. The method used for pharmaceutical processing is based on the Rapid Expansion of supercritical solutions into a liquid SOLVent (RESOLV). We have used this method for the preparation of nanoparticles from various anti-inflammatory (Ibuprofen and Naproxen), anti-fungal (Amphotericin B) and anti-cancer (Paclitaxel) drugs. The anticancer drug nanoparticle formulations were further used for in vitro biological evaluation. In this dissertation, results from the preparation, characterization and biological evaluation of nanoscale drug particles are reported. The effect of experimental conditions on the properties of the nanoparticles prepared via our RESOLV method is discussed. In addition we have used supercritical CO2 as a medium for photoreduction of CO2 using dispersed TiO 2 or metal coated-TiO2 nanoparticles encapsulated in nanoscale cavities of perfluorinated ionomer membranes. These catalytic films are also stable chemically and photochemically, reusable in repeated reactions. The results from the characterization of the nanoparticles and the use of films using different catalysts are presented and discussed.

  14. Several necessary conditions for the evolution of complex forms of life in an artificial environment.

    PubMed

    Suzuki, Hideaki; Ono, Naoaki; Yuta, Kikuo

    2003-01-01

    In order for an artificial life (Alife) system to evolve complex creatures, an artificial environment prepared by a designer has to satisfy several conditions. To clarify this requirement, we first assume that an artificial environment implemented in the computational medium is composed of an information space in which elementary symbols move around and react with each other according to human-prepared elementary rules. As fundamental properties of these factors (space, symbols, transportation, and reaction), we present ten criteria from a comparison with the biochemical reaction space in the real world. Then, in the latter half of the article, we take several computational Alife systems one by one, and assess them in terms of the proposed criteria. The assessment can be used not only for improving previous Alife systems but also for devising new Alife models in which complex forms of artificial creatures can be expected to evolve.

  15. Dual role of starvation signaling in promoting growth and recovery

    PubMed Central

    Leshkowitz, Dena; Barkai, Naama

    2017-01-01

    Growing cells are subject to cycles of nutrient depletion and repletion. A shortage of nutrients activates a starvation program that promotes growth in limiting conditions. To examine whether nutrient-deprived cells prepare also for their subsequent recovery, we followed the transcription program activated in budding yeast transferred to low-phosphate media and defined its contribution to cell growth during phosphate limitation and upon recovery. An initial transcription wave was induced by moderate phosphate depletion that did not affect cell growth. A second transcription wave followed when phosphate became growth limiting. The starvation program contributed to growth only in the second, growth-limiting phase. Notably, the early response, activated at moderate depletion, promoted recovery from starvation by increasing phosphate influx upon transfer to rich medium. Our results suggest that cells subject to nutrient depletion prepare not only for growth in the limiting conditions but also for their predicted recovery once nutrients are replenished. PMID:29236696

  16. The proliferation potential of promastigotes of the main Leishmania species of the old world in NNN culture medium prepared using blood of four different mammals.

    PubMed

    Ladopoulos, Theodoros; Ntais, Pantelis; Tsirigotakis, Nikolaos; Dokianakis, Emmanouil; Antoniou, Maria

    2015-10-01

    The efficacy of the in vitro cultivation of promastigotes of four Leishmania spp. was tested in the biphasic Novy-MacNeal-Nicolle (NNN) medium prepared using blood from different animals (horse, donkey, goat and sheep). The aim was to test which NNN preparation gave the best yield in the shortest time for different parasite species, in order to obtain a large crop of promastigotes for experimental work and for antigen preparation. Promastigotes of Leishmania infantum, Leishmania donovani, Leishmania tropica and Leishmania major, the four main parasite species occurring in the old world, were defrosted from -80 °C and placed, at equal numbers, in the 4 different NNN preparations. At the end of the 7th day, the NNN medium using horse blood produced the greatest number of promastigotes for all Leishmania spp. tested, whilst goat blood proved the poorest medium, providing culture results only for L. infantum. This finding may be explained by the fact that Leishmania is a nicotinamide adenine dinucleotide (NAD) auxotroph and horse erythrocytes support NAD-dependent microorganisms. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Soda-Anthraquinone Durian (Durio Zibethinus Murr.) Rind Linerboard and Corrugated Medium Paper: A Preliminary Test

    NASA Astrophysics Data System (ADS)

    Rizal Masrol, Shaiful; Irwan Ibrahim, Mohd Halim; Adnan, Sharmiza; Mubarak Sa'adon, Amir; Ika Sukarno, Khairil; Fadrol Hisham Yusoff, Mohd

    2017-08-01

    A preliminary test was conducted to investigate the characteristics of linerboard and corrugated medium paper made from durian rind waste. Naturally dried durian rinds were pulped according to Soda-Anthraquinone (Soda-AQ) pulping process with a condition of 20% active alkali, 0.1% AQ, 7:1 liquor to material ratio, 120 minutes cooking time and 170°C cooking temperature. The linerboard and corrugated medium paper with a basis weight of 120 gsm were prepared and evaluated according to Malaysian International Organization for Standardization (MS ISO) and Technical Association of the Pulp and Paper Industry (TAPPI). The results indicate that the characteristics of durian rind linerboard are comparable with other wood or non-wood based paper and current commercial paper. However, low CMT value for corrugated medium and water absorptiveness quality for linerboard could be improved in future. Based on the bulk density (0.672 g/cm3), burst index (3.12 kPa.m2/g) and RCT (2.00 N.m2/g), the durian rind has shown a good potential and suitable as an alternative raw material source for linerboard industry.

  18. Methodological considerations for implementation of lymphocyte subset analysis in a clinical reference laboratory.

    PubMed

    Muirhead, K A; Wallace, P K; Schmitt, T C; Frescatore, R L; Franco, J A; Horan, P K

    1986-01-01

    As the diagnostic utility of lymphocyte subset analysis has been recognized in the clinical research laboratory, a wide variety of reagents and cell preparation, staining and analysis methods have also been described. Methods that are perfectly suitable for analysis of smaller sample numbers in the biological or clinical research setting are not always appropriate and/or applicable in the setting of a high volume clinical reference laboratory. We describe here some of the specific considerations involved in choosing a method for flow cytometric analysis which minimizes sample preparation and data analysis time while maximizing sample stability, viability, and reproducibility. Monoclonal T- and B-cell reagents from three manufacturers were found to give equivalent results for a reference population of healthy individuals. This was true whether direct or indirect immunofluorescence staining was used and whether cells were prepared by Ficoll-Hypaque fractionation (FH) or by lysis of whole blood. When B cells were enumerated using a polyclonal anti-immunoglobulin reagent, less cytophilic immunoglobulin staining was present after lysis than after FH preparation. However, both preparation methods required additional incubation at 37 degrees C to obtain results concordant with monoclonal B-cell reagents. Standard reagents were chosen on the basis of maximum positive/negative separation and the availability of appropriate negative controls. The effects of collection medium and storage conditions on sample stability and reproducibility of subset analysis were also assessed. Specimens collected in heparin and stored at room temperature in buffered medium gave reproducible results for 3 days after specimen collection, using either FH or lysis as the preparation method. General strategies for instrument optimization, quality control, and biohazard containment are also discussed.

  19. Preparation and characterization of poly(acrylic acid)-hydroxyethyl cellulose graft copolymer.

    PubMed

    Abdel-Halim, E S

    2012-10-01

    Poly(acrylic acid) hydroxyethyl cellulose [poly(AA)-HEC] graft copolymer was prepared by polymerizing acrylic acid (AA) with hydroxyethyl cellulose (HEC) using potassium bromate/thiourea dioxide (KBrO(3)/TUD) as redox initiation system. The polymerization reaction was carried out under a variety of conditions including concentrations of AA, KBrO(3) and TUD, material to liquor ratio and polymerization temperature. The polymerization reaction was monitored by withdrawing samples from the reaction medium and measuring the total conversion. The rheological properties of the poly(AA)-HEC graft copolymer were investigated. The total conversion and rheological properties of the graft copolymer depended on the ratio of KBrO(3) to TUD and on acrylic acid concentration as well as temperature and material to liquor ratio. Optimum conditions of the graft copolymer preparation were 30 mmol KBrO(3) and 30 mmol TUD/100g HEC, 100% AA (based on weight of HEC), duration 2h at temperature 50 °C using a material to liquor ratio of 1:10. Copyright © 2012. Published by Elsevier Ltd.

  20. Probing the mechanisms of drug release from amorphous solid dispersions in medium-soluble and medium-insoluble carriers.

    PubMed

    Sun, Dajun D; Lee, Ping I

    2015-08-10

    The objective of the current study is to mechanistically differentiate the dissolution and supersaturation behaviors of amorphous drugs from amorphous solid dispersions (ASDs) based on medium-soluble versus medium-insoluble carriers under nonsink dissolution conditions through a direct head-to-head comparison. ASDs of indomethacin (IND) were prepared in several polymers which exhibit different solubility behaviors in acidic (pH1.2) and basic (pH7.4) dissolution media. The selected polymers range from water-soluble (e.g., PVP and Soluplus) and water-insoluble (e.g., ethylcellulose and Eudragit RL PO) to those only soluble in an acidic or basic dissolution medium (e.g., Eudragit E100, Eudragit L100, and HPMCAS). At 20wt.% drug loading, DSC and powder XRD analysis confirmed that the majority of incorporated IND was present in an amorphous state. Our nonsink dissolution results confirm that whether the carrier matrix is medium soluble determines the release mechanism of amorphous drugs from ASD systems which has a direct impact on the rate of supersaturation generation, thus in turn affecting the evolution of supersaturation in amorphous systems. For example, under nonsink dissolution conditions, the release of amorphous IND from medium-soluble carriers is governed by a dissolution-controlled mechanism leading to an initial surge of supersaturation followed by a sharp decline in drug concentration due to rapid nucleation and crystallization. In contrast, the dissolution of IND ASD from medium-insoluble carriers is more gradual as drug release is regulated by a diffusion-controlled mechanism by which drug supersaturation is built up gradually and sustained over an extended period of time without any apparent decline. Since several tested carrier polymers can be switched from soluble to insoluble by simply changing the pH of the dissolution medium, the results obtained here provide unequivocal evidence of the proposed transition of kinetic solubility profiles from the same ASD system induced by changes in the drug release mechanism in dissolution medium of a different pH. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Exosomes Derived From Pancreatic Stellate Cells: MicroRNA Signature and Effects on Pancreatic Cancer Cells.

    PubMed

    Takikawa, Tetsuya; Masamune, Atsushi; Yoshida, Naoki; Hamada, Shin; Kogure, Takayuki; Shimosegawa, Tooru

    2017-01-01

    Pancreatic stellate cells (PSCs) interact with pancreatic cancer cells in the tumor microenvironment. Cell constituents including microRNAs may be exported from cells within membranous nanovesicles termed exosomes. Exosomes might play a pivotal role in intercellular communication. This study aimed to clarify the microRNA signature of PSC-derived exosomes and their effects on pancreatic cancer cells. Exosomes were prepared from the conditioned medium of immortalized human PSCs. MicroRNAs were prepared from the exosomes and their source PSCs, and the microRNA expression profiles were compared by microarray. The effects of PSC-derived exosomes on proliferation, migration, and the mRNA expression profiles were examined in pancreatic cancer cells. Pancreatic stellate cell-derived exosomes contained a variety of microRNAs including miR-21-5p. Several microRNAs such as miR-451a were enriched in exosomes compared to their source PSCs. Pancreatic stellate cell-derived exosomes stimulated the proliferation, migration and expression of mRNAs for chemokine (C - X - C motif) ligands 1 and 2 in pancreatic cancer cells. The stimulation of proliferation, migration, and chemokine gene expression by the conditioned medium of PSCs was suppressed by GW4869, an exosome inhibitor. We clarified the microRNA expression profile in PSC-derived exosomes. Pancreatic stellate cell-derived exosomes might play a role in the interactions between PSCs and pancreatic cancer cells.

  2. Pressure polymerization of polyester

    DOEpatents

    Maurer, Charles J.; Shaw, Gordon; Smith, Vicky S.; Buelow, Steven J.; Tumas, William; Contreras, Veronica; Martinez, Ronald J.

    2000-08-29

    A process is disclosed for the preparation of a polyester polymer or polyester copolymer under superatmospheric pressure conditions in a pipe or tubular reaction under turbannular flow conditions. Reaction material having a glycol equivalents to carboxylic acid equivalents mole ratio of from 1.0:1 to 1.2:1, together with a superatmospheric dense gaseous medium are fed co-currently to the reactor. Dicarboxylic acid and/or diol raw materials may be injected into any of the reaction zones in the process during operation to achieve the overall desired mole ratio balance. The process operates at temperatures of from about 220.degree. C. to about 320.degree. C., with turbannular flow achieved before the polymer product and gas exit the reactor process. The pressure in the reaction zones can be in the range from 15 psia to 2500 psia. A polymer product having a DP of a greater than 40, more preferably at least about 70, is achieved by the transfer of water from the reacting material polymer melt to the gaseous medium in the reactor.

  3. A Hidden Pitfall in the Preparation of Agar Media Undermines Microorganism Cultivability

    PubMed Central

    Tanaka, Tomohiro; Kawasaki, Kosei; Daimon, Serina; Kitagawa, Wataru; Yamamoto, Kyosuke; Tamaki, Hideyuki; Tanaka, Michiko; Nakatsu, Cindy H.

    2014-01-01

    Microbiologists have been using agar growth medium for over 120 years. It revolutionized microbiology in the 1890s when microbiologists were seeking effective methods to isolate microorganisms, which led to the successful cultivation of microorganisms as single clones. But there has been a disparity between total cell counts and cultivable cell counts on plates, often referred to as the “great plate count anomaly,” that has long been a phenomenon that still remains unsolved. Here, we report that a common practice microbiologists have employed to prepare agar medium has a hidden pitfall: when phosphate was autoclaved together with agar to prepare solid growth media (PT medium), total colony counts were remarkably lower than those grown on agar plates in which phosphate and agar were separately autoclaved and mixed right before solidification (PS medium). We used a pure culture of Gemmatimonas aurantiaca T-27T and three representative sources of environmental samples, soil, sediment, and water, as inocula and compared colony counts between PT and PS agar plates. There were higher numbers of CFU on PS medium than on PT medium using G. aurantiaca or any of the environmental samples. Chemical analysis of PT agar plates suggested that hydrogen peroxide was contributing to growth inhibition. Comparison of 454 pyrosequences of the environmental samples to the isolates revealed that taxa grown on PS medium were more reflective of the original community structure than those grown on PT medium. Moreover, more hitherto-uncultivated microbes grew on PS than on PT medium. PMID:25281372

  4. Production of beta-carotene by a Rhodotorula glutinis mutant in sea water medium.

    PubMed

    Bhosale, P; Gadre, R V

    2001-01-01

    Mutant 32, derived from Rhodotorula glutinis NCIM 3353 produced 76-fold more beta-carotene than the parent strain. In the growth medium prepared in seawater, the total carotenoid content and dry cell mass was 86 mg/l and 16 g/l, respectively, as compared to 70 mg/l and 12 g/l obtained with a medium prepared in distilled water. A 2-fold increase in beta-carotene with simultaneous 2.3-fold decrease in torulene content was also observed. When grown in seawater medium at pH 6.0, 83 +/- 5% carotenoids could be extracted from the cells without any mechanical disintegration.

  5. Formation and Cytotoxicity of Nanoparticles and Nanocubes Prepared from Gold and Silver Salts

    NASA Astrophysics Data System (ADS)

    Banker, Daniel; Dorrell, Skyler; Ivey, Prescott; Scurti, Joseph; Dobbins, Tabbetha

    Photothermal therapy is the use of electromagnetic radiation as the treatment for medical conditions such as cancer. Noble metal nanoparticles and nanocubes are brought to an excited state with laser light and as a result they release vibrational energy in the form of heat, which can be used to kill targeted cancer cells. Wet chemistry gives the basics for the preparation of nanoparticles and nanocubes. Using HAuCl4, AgNO3, tri-sodium citrate and other chemicals, we were able to successfully create gold and silver nanoparticles and nanocubes. The goal is to make sure that 3T3 cells can survive in a nanoparticle or nanocube doped medium so that we can then observe their reaction to photothermal effects. Cell culture techniques were done to 3T3 cells to keep them alive before the testing of cytotoxicity. Photothermal effect refers to the way that our nanoparticles or nanocubes can be photoexcited to release enough heat to kill the cells. We used a UV-Vis spectrophotometer to ensure that the correct wavelength laser. Assuming that the cells will survive living in the doped medium, a medium that has had nanomaterials introduced into it, we will use a high powered laser to observe what the excitation does to the cells since the photothermal effect should result in dead cells.

  6. Buffing, burnishing, and stripping of vinyl asbestos floor tile

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

    Hollett, B.A.; Edwards, A.; Clark, P.J.

    Studies were conducted to evaluate airborne asbestos concentrations during the three principal types of preventative maintenance (low-speed spray-buffing, ultra high-speed burnishing, and wet-stripping) used on asbestos-containing floor tiles. These were done under pre-existing and prepared levels of floor care maintenance. Airborne asbestos concentrations were measured before and during each floor care procedure to determine the magnitude of the increase in airborne asbestos levels during each procedure. Airborne total fiber concentrations were also measured for comparison with the Occupational Safety and Health Administration`s (OSHA) Permissible Exposure Limit (PEL) of 0.1 f/cm{sup 3}. Low-speed spray-buffing and wet-stripping were evaluated on pre-existing floormore » conditions and three levels of prepared floor care conditions (poor, medium, and good). Ultra high-speed burnishing and wet-stripping were evaluated on two levels of prepared floor care conditions (poor and good). Floor care conditions were defined in consultation with the Chemical Specialty Manufacturers Association and other representatives of floor-care chemical manufacturers. Controlled studies were conducted in an unoccupied building at the decommissioned Chanute Air Force Base in Rantoul, Illinois, with the cooperation of the U.S. Air Force. The building offered approximately 8600 ft{sup 2} of open floor space tiled with 9-inch by 9-inch resilient floor tile containing approximately 5% chrysotile asbestos.« less

  7. Hydrogenation of p-chloronitrobenzene on Ni-B Nanometal Catalysts

    NASA Astrophysics Data System (ADS)

    Liu, Yu-Chang; Huang, Chung-Yin; Chen, Yu-Wen

    2006-04-01

    A series of Ni-B catalysts were prepared by mixing nickel acetate in 50% ethanol/water or methanol/water solution. The solution of sodium borohydride (1 M) in excess amount to nickel was then added dropwise into the mixture to ensure full reduction of nickel cations. The mol ratio of boron to nickel in mother solution was 3 to 1. The effects of preparation conditions such as temperature, stirring speed, and sheltering gas on the particle size, surface compositions, electronic states of surface atoms and catalytic activities of the Ni-B catalysts were studied. Ranel nickel catalyst was included for comparison. These catalysts were characterized by N2 sorption, X-ray diffraction, transmission electron microscopy, and X-ray photoelectron spectroscopy. The catalysts were tested for liquid phase hydrogenation of p-chloronitrobenzene. All of the catalysts prepared in this study had nanosized particles. The preparation condition has significant influence on the particle size and surface compositions of the catalyst. The Ni-B catalyst was passivated by boron; therefore it was more stable than Raney nickel and did not catch fire after exposure to air. The catalysts prepared under N2 flow could suppress the oxidation of Ni by the dissolved oxygen in water and had metallic state of nickel. The catalyst prepared with vigorous stirring at 25°C under N2 stream yielded the smallest particles and resulted in the highest activity. It was much more active than the Raney nickel catalyst. The reaction condition also has pronounced effect on the hydrogenation activity. Using methanol as the reaction solvent increased p-chloronitrobenzene conversion to a large extent, compared to that using ethanol as the reaction medium. The selectivity of main product ( p-chloroaniline) was greater than 99% on all of the Ni-B catalysts.

  8. Delayed Administration of Bone Marrow Mesenchymal Stem Cell Conditioned Medium Significantly Improves Outcome After Retinal Ischemia in Rats

    PubMed Central

    Dreixler, John C.; Poston, Jacqueline N.; Balyasnikova, Irina; Shaikh, Afzhal R.; Tupper, Kelsey Y.; Conway, Sineadh; Boddapati, Venkat; Marcet, Marcus M.; Lesniak, Maciej S.; Roth, Steven

    2014-01-01

    Purpose. Delayed treatment after ischemia is often unsatisfactory. We hypothesized that injection of bone marrow stem cell (BMSC) conditioned medium after ischemia could rescue ischemic retina, and in this study we characterized the functional and histological outcomes and mechanisms of this neuroprotection. Methods. Retinal ischemia was produced in adult Wistar rats by increasing intraocular pressure for 55 minutes. Conditioned medium (CM) from rat BMSCs or unconditioned medium (uCM) was injected into the vitreous 24 hours after the end of ischemia. Recovery was assessed 7 days after ischemia using electroretinography, at which time we euthanized the animals and then prepared 4-μm-thick paraffin-embedded retinal sections. TUNEL and Western blot were used to identify apoptotic cells and apoptosis-related gene expression 24 hours after injections; that is, 48 hours after ischemia. Protein content in CM versus uCM was studied using tandem mass spectrometry, and bioinformatics methods were used to model protein interactions. Results. Intravitreal injection of CM 24 hours after ischemia significantly improved retinal function and attenuated cell loss in the retinal ganglion cell layer. CM attenuated postischemic apoptosis and apoptosis-related gene expression. By spectral counting, 19 proteins that met stringent identification criteria were increased in the CM compared to uCM; the majority were extracellular matrix proteins that mapped into an interactional network together with other proteins involved in cell growth and adhesion. Conclusions. By restoring retinal function, attenuating apoptosis, and preventing retinal cell loss after ischemia, CM is a robust means of delayed postischemic intervention. We identified some potential candidate proteins for this effect. PMID:24699381

  9. Fourier transform infrared spectroscopic study of intact cells of the nitrogen-fixing bacterium Azospirillum brasilense

    NASA Astrophysics Data System (ADS)

    Kamnev, A. A.; Ristić, M.; Antonyuk, L. P.; Chernyshev, A. V.; Ignatov, V. V.

    1997-06-01

    The data of Fourier transform infrared (FTIR) spectroscopic measurements performed on intact cells of the soil nitrogen-fixing bacterium Azospirillum brasilense grown in a standard medium and under the conditions of an increased metal uptake are compared and discussed. The structural FTIR information obtained is considered together with atomic absorption spectrometry (AAS) data on the content of metal cations in the bacterial cells. Some methodological aspects concerning preparation of bacterial cell samples for FTIR measurements are also discussed.

  10. The in vitro effects of retrograded starch (resistant starch type 3) from lotus seed starch on the proliferation of Bifidobacterium adolescentis.

    PubMed

    Zhang, Yi; Wang, Ying; Zheng, Baodong; Lu, Xu; Zhuang, Weijing

    2013-11-01

    Prebiotics such as oligosaccharides, fructans, and resistant starch (RS) stimulate the growth of beneficial bacteria in large bowel and modify the human gastrointestinal environment. In this study, compared with glucose (GLU) and high amylose maize starch (HAMS), the in vitro effects of LRS3 and P-LRS3 (RS3 and purified RS3 prepared from lotus seed starch) on the proliferation of bifidobacteria were assessed by assessing the changes in optical density (OD), pH values, short chain fatty acid (SCFA) production, and tolerance ability to gastrointestinal conditions. Significantly higher OD values were obtained from media containing LRS3 and P-LRS3, and especially in the medium containing P-LRS3, the OD value of which reached 1.36 when the concentration of the carbon source was 20 g L(-1). Additionally, the lag phase of bifidobacteria was 8 h in the medium with LRS3 or P-LRS3, whereas it was 16 h in the medium with GLU or HAMS. What is more, a higher content of butyric acid was obtained in the P-LRS3 medium. Compared with GLU and HAMS media, bifidobacteria had a higher tolerance to gastrointestinal conditions in LRS3 and P-LRS3 media. It shows that lotus seed resistant starch, especially P-LRS3, could stimulate the growth of bifidobacteria. The rough surface of resistant starch and the SCFAs produced during fermentation might influence the proliferation of bifidobacteria.

  11. Production of Fumaric Acid in 20-Liter Fermentors

    PubMed Central

    Rhodes, R. A.; Lagoda, A. A.; Misenheimer, T. J.; Smith, M. L.; Anderson, R. F.; Jackson, R. W.

    1962-01-01

    The conditions necessary for the production of fumaric acid in 20-liter fermentors by fermentation of glucose with Rhizopus arrhizus strain NRRL 2582 were determined. Continuous neutralization of fumaric acid was necessary for optimal yields. Yields of the calcium salt were in excess of 65 g of fumaric acid from 100 g of sugar consumed during fermentation of sugar concentrations of 10 to 16%. Conditions established for calcium fumarate production include a simple mineral salts medium, 0.5 v:v:min aeration rate, 300 rev/min agitation rate in a baffled tank, 33 C incubation temperature, CaCO3 to neutralize the acid formed, and a 4 to 5% (v/v) vegetative inoculum. A suitable procedure and medium for the preparation of a vigorous vegetative inoculum were established. The tendency for calcium fumarate fermentations to foam excessively was controlled with a proper antifoam agent added prior to sterilization of the medium and again at daily intervals during fermentation. The production of soluble sodium or potassium fumarates was inhibited when the concentration of fumarates reached 3.5 to 4.0%. No means of overcoming this inhibition was found. Starches and certain other grain-derived carbohydrates were fermented to form calcium fumarate in flask experiments with approximately the same efficiency as was glucose. PMID:16349614

  12. Comparison of Vibrio parahaemolyticus grown in estuarine water and rich medium.

    PubMed Central

    Pace, J; Chai, T J

    1989-01-01

    Cell envelope composition and selected physiological traits of Vibrio parahaemolyticus were studied in regard to the Kanagawa phenomenon and growth conditions. Cell envelopes were prepared from cells cultured in Proteose Peptone-beef extract (Difco Laboratories, Detroit, Mich.) medium or filtered estuarine water. Protein, phospholipid, and lipopolysaccharide contents varied with culture conditions. The phospholipids present in the cell envelopes were identified as phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin. Phosphatidylethanolamine decreased and phosphatidylglycerol increased in cells grown in estuarine water. Profiles of proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated numerous protein species, with four to six predominant proteins ranging from 26,000 to 120,000 in molecular weight. The profile of V. parahaemolyticus cell envelope proteins was unique and might be useful in the identification of the organism. Alkaline phosphatase activity was slightly higher in Kanagawa-negative strains and was higher in cells grown in estuarine water than in cells grown in rich laboratory medium. The DNA levels in estuarine water-grown cells increased, while RNA levels and cell volume decreased. Bacteriophage sensitivity typing demonstrated a close intraspecies relationship. Results indicated that Kanagawa-positive and -negative strains were closely related, but they could be grouped separately and may have undergone starvation-related physiological changes when cultured in estuarine water. Images PMID:2782869

  13. Polyurethane foam loaded with sodium dodecylsulfate for the extraction of 'quat' pesticides from aqueous medium: Optimization of loading conditions.

    PubMed

    Vinhal, Jonas O; Lima, Claudio F; Cassella, Ricardo J

    2016-09-01

    The cationic herbicides paraquat, diquat and difenzoquat are largely used in different cultures worldwide. With this, there is an intrinsic risk of environmental contamination when these herbicides achieve natural waters. The goal of this work was to propose a novel and low-cost sorbent for the removal of the cited herbicides from aqueous medium. The proposed sorbent was prepared by loading polyurethane foam with sodium dodecylsulfate. The influence of several parameters (SDS concentration, HCl concentration and shaking time) on the loading process was investigated. The results obtained in this work demonstrated that all studied variables influenced the loading process, having significant effect on the extraction efficiency of the resulted PUF-SDS. At optimized conditions, the PUF was loaded by shaking 200mg of crushed foam with 200mL of a solution containing 5.0×10(-3)molL(-1) SDS and 0.25molL(-1) HCl, for 30min. The obtained PUF-SDS was efficient for removing the three herbicides from aqueous medium, achieving extraction percentages higher than 90%. The sorption process followed a pseudo second-order kinetics, which presented excellent predictive capacity of the amount of herbicide retained with time. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Quantitative Analysis of the Lamellarity of Giant Liposomes Prepared by the Inverted Emulsion Method

    PubMed Central

    Chiba, Masataka; Miyazaki, Makito; Ishiwata, Shin’ichi

    2014-01-01

    The inverted emulsion method is used to prepare giant liposomes by pushing water-in-oil droplets through the oil/water interface into an aqueous medium. Due to the high encapsulation efficiency of proteins under physiological conditions and the simplicity of the protocol, it has been widely used to prepare various cell models. However, the lamellarity of liposomes prepared by this method has not been evaluated quantitatively. Here, we prepared liposomes that were partially stained with a fluorescent dye, and analyzed their fluorescence intensity under an epifluorescence microscope. The fluorescence intensities of the membranes of individual liposomes were plotted against their diameter. The plots showed discrete distributions, which were classified into several groups. The group with the lowest fluorescence intensity was determined to be unilamellar by monitoring the exchangeability of the inner and the outer solutions of the liposomes in the presence of the pore-forming toxin α-hemolysin. Increasing the lipid concentration dissolved in oil increased the number of liposomes ∼100 times. However, almost all the liposomes were unilamellar even at saturating lipid concentrations. We also investigated the effects of lipid composition and liposome content, such as highly concentrated actin filaments and Xenopus egg extracts, on the lamellarity of the liposomes. Remarkably, over 90% of the liposomes were unilamellar under all conditions examined. We conclude that the inverted emulsion method can be used to efficiently prepare giant unilamellar liposomes and is useful for designing cell models. PMID:25028876

  15. Scedo-Select III: a new semi-selective culture medium for detection of the Scedosporium apiospermum species complex.

    PubMed

    Pham, Trâm; Giraud, Sandrine; Schuliar, Gaëlle; Rougeron, Amandine; Bouchara, Jean-Philippe

    2015-06-01

    The Scedosporium apiospermum complex is responsible for a large variety of infections in human. Members of this complex have become emerging fungal pathogens with an increasing occurrence in patients with underlying conditions such as immunosuppression or cystic fibrosis. A better knowledge of these fungi and of the sources of contamination of the patients is required and more accurate detection methods from the environment are needed. In this context, a highly selective culture medium was developed in the present study. Thus, various aliphatic, cyclic, or aromatic compounds were tested as the sole carbon source, in combination with some inorganic nitrogen sources and fungicides. The best results were obtained with 4-hydroxy-benzoate combined with ammonium sulfate and the fungicides dichloran and benomyl. This new culture medium called Scedo-Select III was shown to support growth of all species of the S. apiospermum complex. Subsequently, this new culture medium was evaluated successfully on water and soil samples, exhibiting higher sensitivity and selectivity than the previously described SceSel+ culture medium. Therefore, this easy-to-prepare and synthetic semi-selective culture medium may be useful to clarify the ecology of these fungi and to identify their reservoirs in patients' environment. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  16. Pseudo-thermosetting chitosan hydrogels for biomedical application.

    PubMed

    Berger, J; Reist, M; Chenite, A; Felt-Baeyens, O; Mayer, J M; Gurny, R

    2005-01-20

    To prepare transparent chitosan/beta-glycerophosphate (betaGP) pseudo-thermosetting hydrogels, the deacetylation degree (DD) of chitosan has been modified by reacetylation with acetic anhydride. Two methods (I and II) of reacetylation have been compared and have shown that the use of previously filtered chitosan, dilution of acetic anhydride and reduction of temperature in method II improves efficiency and reproducibility. Chitosans with DD ranging from 35.0 to 83.2% have been prepared according to method II under homogeneous and non-homogeneous reacetylation conditions and the turbidity of chitosan/betaGP hydrogels containing homogeneously or non-homogeneously reacetylated chitosan has been investigated. Turbidity is shown to be modulated by the DD of chitosan and by the homogeneity of the medium during reacetylation, which influences the distribution mode of the chitosan monomers. The preparation of transparent chitosan/betaGP hydrogels requires a homogeneously reacetylated chitosan with a DD between 35 and 50%.

  17. Pseudo-thermosetting chitosan hydrogels for biomedical application.

    PubMed

    Berger, J; Reist, M; Chenite, A; Felt-Baeyens, O; Mayer, J M; Gurny, R

    2005-01-06

    To prepare transparent chitosan/beta-glycerophosphate (betaGP) pseudo-thermosetting hydrogels, the deacetylation degree (DD) of chitosan has been modified by reacetylation with acetic anhydride. Two methods (I and II) of reacetylation have been compared and have shown that the use of previously filtered chitosan, dilution of acetic anhydride and reduction of temperature in method II improves efficiency and reproducibility. Chitosans with DD ranging from 35.0 to 83.2% have been prepared according to method II under homogeneous and non-homogeneous reacetylation conditions and the turbidity of chitosan/betaGP hydrogels containing homogeneously or non-homogeneously reacetylated chitosan has been investigated. Turbidity is shown to be modulated by the DD of chitosan and by the homogeneity of the medium during reacetylation, which influences the distribution mode of the chitosan monomers. The preparation of transparent chitosan/betaGP hydrogels requires a homogeneously reacetylated chitosan with a DD between 35 and 50%.

  18. Palladium-Catalyzed SN2'-Cyclization of Ambivalent (Bromoalkadienyl)malonates: Preparation of Medium- to Large-Membered Endocyclic Allenes.

    PubMed

    Ichio, Hiroaki; Murakami, Hidetoshi; Chen, Yen-Chou; Takahashi, Tamotsu; Ogasawara, Masamichi

    2017-07-21

    A palladium-catalyzed reaction for preparing various endocyclic allenes was developed. The substrates for the reaction were readily available ω-(pronucleophile-tethered)-3-bromo-1,3-alkadienes, and a palladium-catalyst facilitated their unimolecular S N 2'-cyclization in the presence of potassium tert-butoxide to give the corresponding 9- to 16-membered endocyclic allenes in fair yields of up to 67% together with the dimeric 16- to 32-membered endocyclic bis-allenes and other oligomeric/polymeric intermolecular reaction products. For higher yields of the monomeric endocyclic allenes, the reaction needed to be conducted under high-dilution conditions. Using a chiral palladium catalyst, axially chiral endocyclic allenes were obtained in up to 70% ee.

  19. Intestinal myiasis.

    PubMed

    Udgaonkar, U S; Dharamsi, R; Kulkarni, S A; Shah, S R; Patil, S S; Bhosale, A L; Gadgil, S A; Mohite, R S

    2012-01-01

    Intestinal myiasis is a condition when the fly larvae inhabit the gastrointestinal tract and are passed out in faeces. This type of infestation results when eggs or larvae of the fly, deposited on food are inadvertently taken by man. They survive the unfavourable conditions within the gastrointestinal tract and produce disturbances, which may vary from mild to severe. The condition is not uncommon and is often misdiagnosed as pinworm infestation. Correct diagnosis by the clinical microbiologist is important to avoid unnecessary treatment. We had 7 cases of intestinal myiasis. In 2 cases the larvae were reared to adult fly in modified meat and sand medium (developed by Udgaonkar). This medium is simple and can be easily prepared in the laboratory. Of the 7 larvae, 5 were Sarcophaga haemorrhoidalis, 1 Megaselia species and 1 was identified as Muscina stabulans. S. haemorrhoidalis was the commonest maggot involved. A high index of suspicion is required for clinical diagnosis when the patient complains of passing wriggling worms in faeces for a long period without any response to antihelminthics. The reason for long duration of illness and recurrence of infestation is baffling. The nearest to cure was colonic wash. We feel prevention is of utmost importance, which is to avoid eating food articles with easy access to flies.

  20. Human Induced Hepatic Lineage-Oriented Stem Cells: Autonomous Specification of Human iPS Cells toward Hepatocyte-Like Cells without Any Exogenous Differentiation Factors

    PubMed Central

    Yanagi, Satoshi; Kato, Chika; Takashima, Ryokichi; Kobayashi, Eiji; Hagiwara, Keitaro; Ochiya, Takahiro

    2015-01-01

    Preparing targeted cells for medical applications from human induced pluripotent stem cells (hiPSCs) using growth factors, compounds, or gene transfer has been challenging. Here, we report that human induced hepatic lineage-oriented stem cells (hiHSCs) were generated and expanded as a new type of hiPSC under non-typical coculture with feeder cells in a chemically defined hiPSC medium at a very high density. Self-renewing hiHSCs expressed markers of both human embryonic stem cells (hESCs) and hepatocytes. Those cells were highly expandable, markedly enhancing gene expression of serum hepatic proteins and cytochrome P450 enzymes with the omission of FGF-2 from an undefined hiPSC medium. The hepatic specification of hiHSCs was not attributable to the genetic and epigenetic backgrounds of the starting cells, as they were established from distinct donors and different types of cells. Approximately 90% of hiHSCs autonomously differentiated to hepatocyte-like cells, even in a defined minimum medium without any of the exogenous growth factors necessary for hepatic specification. After 12 days of this culture, the differentiated cells significantly enhanced gene expression of serum hepatic proteins (ALB, SERPINA1, TTR, TF, FABP1, FGG, AGT, RBP4, and AHSG), conjugating enzymes (UGT2B4, UGT2B7, UGT2B10, GSTA2, and GSTA5), transporters (SULT2A1, SLC13A5, and SLCO2B1), and urea cycle-related enzymes (ARG1 and CPS1). In addition, the hepatocyte-like cells performed key functions of urea synthesis, albumin secretion, glycogen storage, indocyanine green uptake, and low-density lipoprotein uptake. The autonomous hepatic specification of hiHSCs was due to their culture conditions (coculture with feeder cells in a defined hiPSC medium at a very high density) in self-renewal rather than in differentiation. These results suggest the feasibility of preparing large quantities of hepatocytes as a convenient and inexpensive hiPSC differentiation. Our study also suggests the necessity of optimizing culture conditions to generate other specific lineage-oriented hiPSCs, allowing for a very simple differentiation. PMID:25875613

  1. Medium chain triglycerides (MCT) formulas in paediatric and allergological practice.

    PubMed

    Łoś-Rycharska, Ewa; Kieraszewicz, Zuzanna; Czerwionka-Szaflarska, Mieczysława

    2016-01-01

    Fats constitute the most significant nutritional source of energy. Their proper use by the body conditions a number of complex mechanisms of digestion, absorption, distribution, and metabolism. These mechanisms are facilitated by fats made of medium chain fatty acids; therefore, they are an easy and quick source of energy. Thus, an increased supply of medium chain triglycerides (MCT) is particularly important in patients with disturbances of digestion and absorption such as disturbed bile secretion, classic coeliac disease, short bowel syndrome, inflammatory diseases of the intestines, disturbed outflow of lymph, some metabolic disease, and severe food allergies, as well as in prematurely born neonates. Use of preparations containing an additive of MCT is limited, especially if they are to be used for a longer period of time. With a large quantity of MCT in a diet, there is a risk of deficiency of necessary unsaturated fatty acids and some fat-soluble vitamins. The caloricity of MTC compared to long-chain triglycerides is lower, and formulas with MCT are characterised by higher osmolality. Medium chain triglycerides is not recommended as an additive to standard formulas for healthy children. The use of MCT should be limited to strictly specified medical indications.

  2. Medium chain triglycerides (MCT) formulas in paediatric and allergological practice

    PubMed Central

    Łoś-Rycharska, Ewa; Kieraszewicz, Zuzanna

    2016-01-01

    Fats constitute the most significant nutritional source of energy. Their proper use by the body conditions a number of complex mechanisms of digestion, absorption, distribution, and metabolism. These mechanisms are facilitated by fats made of medium chain fatty acids; therefore, they are an easy and quick source of energy. Thus, an increased supply of medium chain triglycerides (MCT) is particularly important in patients with disturbances of digestion and absorption such as disturbed bile secretion, classic coeliac disease, short bowel syndrome, inflammatory diseases of the intestines, disturbed outflow of lymph, some metabolic disease, and severe food allergies, as well as in prematurely born neonates. Use of preparations containing an additive of MCT is limited, especially if they are to be used for a longer period of time. With a large quantity of MCT in a diet, there is a risk of deficiency of necessary unsaturated fatty acids and some fat-soluble vitamins. The caloricity of MTC compared to long-chain triglycerides is lower, and formulas with MCT are characterised by higher osmolality. Medium chain triglycerides is not recommended as an additive to standard formulas for healthy children. The use of MCT should be limited to strictly specified medical indications. PMID:28053676

  3. Qualitative and quantitative measures of various compounded formulations of 17-hydroxyprogesterone caproate

    PubMed Central

    CARITIS, Steve N.; ZHAO, Yang; BETTINGER, Joseph; VENKATARAMANAN, Raman

    2013-01-01

    Objectives 17-hydroxyprogesterone caproate (17-OHPC) is available both as an FDA – approved medication and as a product prepared for individual patients by compounding pharmacies. Compounding pharmacies may omit the preservative used in the FDA approved formulation or use an alternate preservative and may dispense 17-OHPC in containers that differ from the FDA-approved product. The objective of this study was to assess the stability, microbiological and pyrogen status of 17-OHPC formulations under various compounding and dispensing conditions. Methods 17-OHPC was prepared by a local compounding pharmacy. The formulations prepared included one identical to the FDA approved product with benzyl alcohol as a preservative, one with benzalkonium chloride as a preservative and one without a preservative. These various formulations were dispensed into either single dose 1 ml plastic syringes or glass vials or 10 ml glass vials. Concentration of 17-OHPC and microbial and pyrogen status were evaluated at various time intervals over the ensuing 19 weeks. Results The concentration of 17-OHPC did not change over the duration of study regardless of the dispensing medium used or the absence or presence of any preservatives. The preparations remained microbe and pyrogen free during the study period regardless of the dispensing medium used or the absence of presence of any preservatives. Conclusions Products containing 17-OPHC tested in this study were quite stable over the 19-week period of study in different dispensing containers and in absence or presence of a different preservative. The compounded products remained sterile and pyrogen free during the period of observation. PMID:23453884

  4. Semi-preparative scale purification of enterococcal bacteriocin enterocin EJ97, and evaluation of substrates for its production.

    PubMed

    López, Rosario Lucas; García, Ma Teresa; Abriouel, Hikmate; Ben Omar, Nabil; Grande, Ma José; Martínez-Cañamero, Magdalena; Gálvez, Antonio

    2007-12-01

    The influence of substrate composition on the production of enterocin EJ97 and the conditions for semi-preparative bacteriocin recovery have been studied. Final bacteriocin concentrations of 12.5 or 15.6 mg/l were obtained in the commercial media brain heart infusion broth (BHI) and tryptic soya broth, respectively. The bacteriocin was also produced in the complex medium CM (8.75 mg/l), in which the vitamin supplement was essential for production. Some combinations of meat peptone and yeast extract plus either soy peptone or BHI also supported bacteriocin production, at concentrations of 6.25-7.5 mg/l. In cow milk (whole, half-skimmed, and skimmed), the final bacteriocin concentrations obtained ranged from 7.5 to 11.25 mg/l. Highest bacteriocin activity was obtained by using pasteurised milk whey as growth substrate (up to 25 mg/l), suggesting that this bacteriocin can be obtained on a large scale by using this cheap food-grade industrial by-product. Highest bacteriocin titres were always obtained after 8 h of incubation at 37 degrees C. Semi-preparative concentration and purification of enterocin EJ97 produced in a complex medium was achieved by bulk cation exchange chromatography without previous cell separation, followed by reversed-phase chromatography. This two-step procedure allowed preparation of milligram quantities of purified bacteriocin, which is an improvement compared to purification procedures established for most other bacteriocins (35). The availability of purified enterocin EJ97 will facilitate other studies such as the elucidation of its molecular structure and its interaction with target bacteria.

  5. Amyloid-β(1–42) Protofibrils Formed in Modified Artificial Cerebrospinal Fluid Bind and Activate Microglia

    PubMed Central

    Paranjape, Geeta S.; Terrill, Shana E.; Gouwens, Lisa K.; Ruck, Benjamin M.; Nichols, Michael R.

    2012-01-01

    Soluble aggregated forms of amyloid-β protein (Aβ) have garnered significant attention recently for their role in Alzheimer’s disease (AD). Protofibrils are a subset of these soluble species and are considered intermediates in the aggregation pathway to mature Aβ fibrils. Biological studies have demonstrated that protofibrils exhibit both toxic and inflammatory activities. It is important in these in vitro studies to prepare protofibrils using solution conditions that are appropriate for cellular studies as well as conducive to biophysical characterization of protofibrils. Here we describe the preparation and characterization of Aβ(1–42) protofibrils in modified artificial cerebrospinal fluid (aCSF) and demonstrate their prominent binding and activation of microglial cells. A simple phosphate/bicarbonate buffer system was prepared that maintained the ionic strength and cell compatibility of F-12 medium but did not contain numerous supplements that interfere with spectroscopic analyses of Aβ protofibrils. Reconstitution of Aβ(1–42) in aCSF and isolation with size exclusion chromatography (SEC) revealed curvilinear β-sheet protofibrils <100 nm in length and hydrodynamic radii of 21 nm. Protofibril concentration determination by BCA assay, which was not possible in F-12 medium, was more accurately measured in aCSF. Protofibrils formed and isolated in aCSF, but not monomers, markedly stimulated TNFα production in BV-2 and primary microglia and bound in significant amounts to microglial membranes. This report demonstrates the suitability of a modified aCSF system for preparing SEC-isolated Aβ(1–42) protofibrils and underscores the unique ability of protofibrils to functionally interact with microglia. PMID:23242692

  6. Enhanced biocatalytic esterification with lipase-immobilized chitosan/graphene oxide beads.

    PubMed

    Lau, Siaw Cheng; Lim, Hong Ngee; Basri, Mahiran; Fard Masoumi, Hamid Reza; Ahmad Tajudin, Asilah; Huang, Nay Ming; Pandikumar, Alagarsamy; Chia, Chin Hua; Chia, Chi Hua; Andou, Yoshito

    2014-01-01

    In this work, lipase from Candida rugosa was immobilized onto chitosan/graphene oxide beads. This was to provide an enzyme-immobilizing carrier with excellent enzyme immobilization activity for an enzyme group requiring hydrophilicity on the immobilizing carrier. In addition, this work involved a process for the preparation of an enzymatically active product insoluble in a reaction medium consisting of lauric acid and oleyl alcohol as reactants and hexane as a solvent. This product enabled the stability of the enzyme under the working conditions and allowed the enzyme to be readily isolated from the support. In particular, this meant that an enzymatic reaction could be stopped by the simple mechanical separation of the "insoluble" enzyme from the reaction medium. Chitosan was incorporated with graphene oxide because the latter was able to enhance the physical strength of the chitosan beads by its superior mechanical integrity and low thermal conductivity. The X-ray diffraction pattern showed that the graphene oxide was successfully embedded within the structure of the chitosan. Further, the lipase incorporation on the beads was confirmed by a thermo-gravimetric analysis. The lipase immobilization on the beads involved the functionalization with coupling agents, N-hydroxysulfosuccinimide sodium (NHS) and 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC), and it possessed a high enzyme activity of 64 U. The overall esterification conversion of the prepared product was 78% at 60 °C, and it attained conversions of 98% and 88% with commercially available lipozyme and novozyme, respectively, under similar experimental conditions.

  7. Enhanced Biocatalytic Esterification with Lipase-Immobilized Chitosan/Graphene Oxide Beads

    PubMed Central

    Lau, Siaw Cheng; Lim, Hong Ngee; Basri, Mahiran; Fard Masoumi, Hamid Reza; Ahmad Tajudin, Asilah; Huang, Nay Ming; Pandikumar, Alagarsamy; Chia, Chi Hua; Andou, Yoshito

    2014-01-01

    In this work, lipase from Candida rugosa was immobilized onto chitosan/graphene oxide beads. This was to provide an enzyme-immobilizing carrier with excellent enzyme immobilization activity for an enzyme group requiring hydrophilicity on the immobilizing carrier. In addition, this work involved a process for the preparation of an enzymatically active product insoluble in a reaction medium consisting of lauric acid and oleyl alcohol as reactants and hexane as a solvent. This product enabled the stability of the enzyme under the working conditions and allowed the enzyme to be readily isolated from the support. In particular, this meant that an enzymatic reaction could be stopped by the simple mechanical separation of the “insoluble” enzyme from the reaction medium. Chitosan was incorporated with graphene oxide because the latter was able to enhance the physical strength of the chitosan beads by its superior mechanical integrity and low thermal conductivity. The X-ray diffraction pattern showed that the graphene oxide was successfully embedded within the structure of the chitosan. Further, the lipase incorporation on the beads was confirmed by a thermo-gravimetric analysis. The lipase immobilization on the beads involved the functionalization with coupling agents, N-hydroxysulfosuccinimide sodium (NHS) and 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC), and it possessed a high enzyme activity of 64 U. The overall esterification conversion of the prepared product was 78% at 60°C, and it attained conversions of 98% and 88% with commercially available lipozyme and novozyme, respectively, under similar experimental conditions. PMID:25127038

  8. Gold nanoparticles prepared by electro-exploding wire technique in aqueous solutions

    NASA Astrophysics Data System (ADS)

    Kumar, Lalit; Kapoor, Akanksha; Meghwal, Mayank; Annapoorni, S.

    2016-05-01

    This article presents an effective approach for the synthesis of Au nanoparticles via an environmentally benevolent electro-exploding wire (EEW) technique. In this process, Au nanoparticles evolve through the plasma generated from the parent Au metal. Compared to other typical chemical methods, electro-exploding wire technique is a simple and economical technique which normally operates in water or organic liquids under ambient conditions. Efficient size control was achieved using different aqueous medium like (1mM) NaCl, deionized water and aqueous solution of sodium hydroxide (NaOH, pH 9.5) using identical electro-exploding conditions. The gold nanoparticles exhibited the UV-vis absorption spectrum with a maximum absorption band at 530 nm, similar to that of gold nanoparticles chemically prepared in a solution. The mechanism of size variation of Au nanoparticles is also proposed. The results obtained help to develop methodologies for the control of EEW based nanoparticle growth and the functionalization of nanoparticle surfaces by specific interactions.

  9. [Defective function of bioindicators for steam sterilization].

    PubMed

    Botzenhart, K; Merkt-Kinzler, M

    1990-05-01

    It can be shown, that under certain conditions commercially available indicators with Bacillus stearothermophilus and packages of native spores from soil prepared according to DIN 58 946/4 react differently to treatment in a lab-type steam sterilizer. The differences were most evident when incomplete evacuation of air had to be supposed. These results lead to the conclusion that some bioindicators are not able to show the inefficient function of steam sterilizers caused by local residuals of air. This may be caused by the properties of the selected strain, by the conditions of growth and preparation of the spores and by the culture medium used after exposition. The results of our experiments as well as the resistance of mesophilic spore forming bacilli against dry heat described by other authors make it necessary to test the resistance of bioindicators for steam sterilization not only against steam according to DIN 58946/4 but also against dry heat or mixtures of steam and air.

  10. Effect of Processing Variables on Tensile Modulus and Morphology of Polyethylene/Clay Nanocomposites Prepared in an Internal Mixer

    NASA Astrophysics Data System (ADS)

    Ujianto, O.; Jollands, M.; Kao, N.

    2018-03-01

    A comparative study on effect of internal mixer on high density Polyethylene (HDPE)/clay nanocomposites preparation was done. Effect of temperature, rotor rotation (rpm), and mixing time, as well as rotor type (Roller and Banbury) on mechanical properties and morphology of HDPE/clay nanocomposites were studied using Box-Behnken experimental design. The model was developed according to secant modulus and confirmed to morphology analysis using Transmission Electron Microscopy (TEM). The finding suggests that there is different mechanisms occurred in each rotor to improve the mechanical properties. The mechanism in Roller is medium shear and medium diffusion, while Banbury is high shear and low diffusion. The difference in mechanism to disperse the clay particles attribute to the different optimum processing conditions in each rotor. The settings for roller samples are predicted around mid temperature, mid speed, and mid mixing time. There is no optimum setting for Banbury within the processing boundaries. The best settings for Banbury are at low, high, low settings. The morphology results showed a hybrid composite structure, with some exfoliations and some intercalations. There was a correlation between better mechanical properties and morphology with more exfoliation and thinner intercalated particles.

  11. Effects of laser fluence and liquid media on preparation of small Ag nanoparticles by laser ablation in liquid

    NASA Astrophysics Data System (ADS)

    Moura, Caroline Gomes; Pereira, Rafael Santiago Floriani; Andritschky, Martin; Lopes, Augusto Luís Barros; Grilo, João Paulo de Freitas; Nascimento, Rubens Maribondo do; Silva, Filipe Samuel

    2017-12-01

    This study aims to assess a method for preparation of small and highly stable Ag nanoparticles by nanosecond laser ablation in liquid. Effect of liquid medium and laser fluence on the size, morphology and structure of produced nanoparticles has been studied experimentally. Pulses of a Nd:YAG laser of 1064 nm wavelength at 35 ns pulse width at different fluences were employed to irradiate the silver target in different environments (water, ethanol and acetone). The UV-Visible absorption spectra of nanoparticles exhibit surface plasmon resonance absorption peak in the UV region. STEM and TEM micrographs were used to evaluate the size and shape of nanoparticles. The stability of silver colloids in terms of oxidation at different liquid media was analyzed by SAED patterns. The results showed that characteristics of Ag nanoparticles and their production rate were strongly influenced by varying laser fluence and liquid medium. Particles from 2 to 80 nm of diameter were produced using different conditions and no oxidation was found in ethanol and acetone media. This work puts in evidence a promising approach to produce small nanoparticles by using high laser fluence energy.

  12. Selective medium for aerobic incubation of Campylobacter

    USDA-ARS?s Scientific Manuscript database

    Studies were conducted on the formulation of a selective medium that could be used to isolate Campylobacter from mixed bacterial cultures using aerobic incubation. A non-selective, basal broth medium was prepared and supplemented with Bolton, Cefex, or Skirrow antibiotic mixtures. The ability of pur...

  13. A FORTRAN program for calculating nonlinear seismic ground response

    USGS Publications Warehouse

    Joyner, William B.

    1977-01-01

    The program described here was designed for calculating the nonlinear seismic response of a system of horizontal soil layers underlain by a semi-infinite elastic medium representing bedrock. Excitation is a vertically incident shear wave in the underlying medium. The nonlinear hysteretic behavior of the soil is represented by a model consisting of simple linear springs and Coulomb friction elements arranged as shown. A boundary condition is used which takes account of finite rigidity in the elastic substratum. The computations are performed by an explicit finite-difference scheme that proceeds step by step in space and time. A brief program description is provided here with instructions for preparing the input and a source listing. A more detailed discussion of the method is presented elsewhere as is the description of a different program employing implicit integration.

  14. Maize Inoculation with Azospirillum brasilense Ab-V5 Cells Enriched with Exopolysaccharides and Polyhydroxybutyrate Results in High Productivity under Low N Fertilizer Input

    PubMed Central

    Oliveira, André L. M.; Santos, Odair J. A. P.; Marcelino, Paulo R. F.; Milani, Karina M. L.; Zuluaga, Mónica Y. A.; Zucareli, Claudemir; Gonçalves, Leandro S. A.

    2017-01-01

    Although Azospirillum strains used in commercial inoculant formulations presents diazotrophic activity, it has been reported that their ability to produce phytohormones plays a pivotal role in plant growth-promotion, leading to a general recommendation of its use in association with regular N-fertilizer doses. In addition, a high variability in the effectiveness of Azospirillum inoculants is still reported under field conditions, contributing to the adoption of the inoculation technology as an additional management practice rather than its use as an alternative practice to the use of chemical inputs in agriculture. To investigate whether the content of stress-resistance biopolymers would improve the viability and performance of Azospirillum inoculants when used as substitute of N-fertilizers, biomass of A. brasilense strain Ab-V5 enriched in exopolysaccharides (EPS) and polyhydroxybutirate (PHB) was produced using a new culture medium developed by factorial mixture design, and the effectiveness of resulting inoculants was evaluated under field conditions. The culture medium formulation extended the log phase of A. brasilense cultures, which presented higher cell counts and increased EPS and PHB contents than observed in the cultures grown in the OAB medium used as control. An inoculation trial with maize conducted under greenhouse conditions and using the biopolymers-enriched Ab-V5 cells demonstrated the importance of EPS and PHB to the long term bacterial viability in soil and to the effectiveness of inoculation. The effectiveness of liquid and peat inoculants prepared with Ab-V5 cells enriched with EPS and PHB was also evaluated under field conditions, using maize as target crop along different seasons, with the inoculants applied directly over seeds or at topdressing under limiting levels of N-fertilization. No additive effect on yield resulted from inoculation under high N fertilizer input, while inoculated plants grown under 80% reduction in N fertilizer showed yields at levels compared to fully fertilized plants, regardless the inoculation method. The presented data highlights the feasibility to partially substitute the N-fertilizer demand in non-legume crops using high-quality inoculant formulations, prepared with diazotrophic bacteria enriched with stress-resistance biopolymers that confer increased viability an effectiveness to the bacterial cells. PMID:29018432

  15. Development of sol-gel bioactive glass for hard tissue regeneration

    NASA Astrophysics Data System (ADS)

    Noor, Siti Noor Fazliah Mohd; Zain, Nurul Shazwani Mohd; Wei, Poh Yong; Azizan, Nur Syazana; Mohamad, Hasmaliza

    2016-12-01

    The regeneration of hard tissues requires various contributing factors such as cells, scaffolds and growth factors. Bioactive glasses are known for its properties to stimulate hard tissue regeneration. In this study, sol-gel bioactive glasses (BG) were prepared and characterized. Sol-gel BG powders having particle size less than 25 µm were incubated with cell culture medium for 4 hours at 37°C on continuous rolling, and then the medium was filtered using 0.22 µm syringe filters. Prior to use, the SGBG-conditioned media were supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) antibiotic-antimycotic, and were allowed to equilibrate overnight inside a CO2 incubator. The human dental pulp stem cells (DPSC) were incubated with the BG-conditioned media and their viability and proliferation were assessed at day 1, 2, 4 and 7 using Alamar Blue and MTT assays. The results showed that BG at various powders to liquid ratio concentrations promoted DPSC growth. The BG have potential to be used for hard tissue regeneration especially in the field of regenerative dentistry.

  16. [Survival of probiotic microorganisms in the conditions in vitro imitating the process of human digestion].

    PubMed

    Darmov, I V; Chicherin, I Iu; Pogorel'skiĭ, I P; Lundovskikh, I A

    2011-01-01

    Assessment of survival bifidobacteria and lactobacteria under the conditions in vitro, simulating digestion in human stomach and intestine, and study of survival probiotic and indigenous microorganisms in co-cultivation on solid nutrient medium. Probiotic microorganisms from commercial preparations Bifidobacterin and Lactobacterin, clinical isolates lactobacillus (Lactobacillus acidophilus No 1, L. brevis No 2) were used in experiments. Survival study of probiotic microorganisms was performed on a model in vitro, simulating the process of digestion in the human body. Assessment of the relationship of probiotic microorganisms and indigenous microorganisms was carried out in co-cultivation in vitro on solid nutrient medium. A significant reduction in the number of viable probiotic microorganisms during their incubation in model media was set as well as suppression of probiotic microorganisms growth by cultures of a clinical strains of lactobacillus, corresponding to biocompatibility by type "host against probiotic". While choosing probiotics in the treatment of dysbacterioses the character of relationship between probiotic microorganisms and indigenous microorganisms of a patient is recommended to be preliminarily tested. Also microorganisms of own microflora should be stimulated using modern prebiotics.

  17. Fabrication of magnetic biochar as a treatment medium for As(V) via pyrolysis of FeCl3-pretreated spent coffee ground.

    PubMed

    Cho, Dong-Wan; Yoon, Kwangsuk; Kwon, Eilhann E; Biswas, Jayanta Kumar; Song, Hocheol

    2017-10-01

    This study investigated the preparation of magnetic biochar from N 2 - and CO 2 -assisted pyrolysis of spent coffee ground (SCG) for use as an adsorption medium for As(V), and the effects of FeCl 3 pretreatment of SCG on the material properties and adsorption capability of the produced biochar. Pyrolysis of FeCl 3 -pretreated SCG in CO 2 atmosphere produced highly porous biochar with its surface area ∼70 times greater than that produced in N 2 condition. However, despite the small surface area, biochar produced in N 2 showed greater As(V) adsorption capability. X-ray diffraction and X-ray photoelectron spectrometer analyses identified Fe 3 C and Fe 3 O 4 as dominant mineral phases in N 2 and CO 2 conditions, with the former being much more adsorptive toward As(V). The overall results suggest functional biochar can be facilely fabricated by necessary pretreatment to expand the applicability of biochar for specific purposes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Effects of chitosan molecular weight on the physical and dissolution characteristics of amorphous curcumin-chitosan nanoparticle complex.

    PubMed

    Yu, Hong; Nguyen, Minh-Hiep; Hadinoto, Kunn

    2018-01-01

    To investigate the effects of varying molecular weight (MW) of chitosan (CHI) used in the complexation with curcumin (CUR) on the physical and dissolution characteristics of the amorphous CUR-CHI nanoparticle complex produced. Amorphous CUR-CHI nanoparticle complex (or CUR nanoplex in short) recently emerged as a promising bioavailability enhancement strategy of CUR attributed to its fast dissolution, supersaturation generation capability, and simple preparation. Existing CUR nanoplex prepared using low MW CHI, however, exhibited poor colloidal stability during storage. Herein we hypothesized that the colloidal stability could be improved by using CHI of higher MW. The effects of this approach on the nanoplex's other characteristics were simultaneously investigated. The CUR nanoplex was prepared by electrostatically driven self-assembled complexation between CUR and oppositely charged CHI of three different MWs (i.e. low, medium, and high). Besides colloidal stability, the effects of MW variation were investigated for the nanoplex's (1) other physical characteristics (i.e. size, zeta potential, CUR payload, amorphous state stability), (2) preparation efficiency (i.e. CUR utilization rate, yield), and (3) dissolutions under sink condition and supersaturation generation. CUR nanoplex prepared using CHI of high MW exhibited improved colloidal stability, larger size, superior morphology, and prolonged supersaturation generation. On the other hand, the effects of MW variation on the payload, amorphous state stability, preparation efficiency, and dissolution under sink condition were found to be insignificant. Varying MW of CHI used was an effective means to improve certain aspects of the CUR nanoplex characteristics with minimal adverse effects on the others.

  19. New on-line separation workflow of microbial metabolites via hyphenation of analytical and preparative comprehensive two-dimensional liquid chromatography.

    PubMed

    Yan, Xia; Wang, Li-Juan; Wu, Zhen; Wu, Yun-Long; Liu, Xiu-Xiu; Chang, Fang-Rong; Fang, Mei-Juan; Qiu, Ying-Kun

    2016-10-15

    Microbial metabolites represent an important source of bioactive natural products, but always exhibit diverse of chemical structures or complicated chemical composition with low active ingredients content. Traditional separation methods rely mainly on off-line combination of open-column chromatography and preparative high performance liquid chromatography (HPLC). However, the multi-step and prolonged separation procedure might lead to exposure to oxygen and structural transformation of metabolites. In the present work, a new two-dimensional separation workflow for fast isolation and analysis of microbial metabolites from Chaetomium globosum SNSHI-5, a cytotoxic fungus derived from extreme environment. The advantage of this analytical comprehensive two-dimensional liquid chromatography (2D-LC) lies on its ability to analyze the composition of the metabolites, and to optimize the separation conditions for the preparative 2D-LC. Furthermore, gram scale preparative 2D-LC separation of the crude fungus extract could be performed on a medium-pressure liquid chromatograph×preparative high-performance liquid chromatography system, under the optimized condition. Interestingly, 12 cytochalasan derivatives, including two new compounds named cytoglobosin Ab (3) and isochaetoglobosin Db (8), were successfully obtained with high purity in a short period of time. The structures of the isolated metabolites were comprehensively characterized by HR ESI-MS and NMR. To be highlighted, this is the first report on the combination of analytical and preparative 2D-LC for the separation of microbial metabolites. The new workflow exhibited apparent advantages in separation efficiency and sample treatment capacity compared with conventional methods. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Preparation of tritium-labeled optical isomers of amino acids by ligand exchange chromatography on polyacrylamide sorbent containing L-phenylalanine groupings

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

    Zolotarev, Yu.A.; Penkina, V.I.; Dostavalov, I.N.

    Tritium-labeled optically active amino acids are obtained by resolving racemates of the corresponding amino acids by chromatography on a chiral polyacrylamide sorbent, filled with copper ions. The chiral sorbent is obtained by the action of formaldehyde and L-phenylalanine on a Biogel P-4 polyacrylamide gel in an alkaline medium. Data are given on the ligand exchange chromatography of amino acids on this sorbent, depending on the degree of filling of the sorbent by copper ions and the concentration of the eluent. Conditions were selected for the quantitative resolution of racemates of amino acids and examples are given of a preparative obtainingmore » of tritium labeled optical isomers of amino acids.« less

  1. Anaerobic utilization of phosphite/phosphine as a sole source of phosphorus: implication to growth in the Jovian environment.

    PubMed

    Foster, T L; Winans, L

    1977-01-01

    The objective of the investigation was to isolate anaerobic micro-organisms which had the ability to utilize inorganic phosphorus in forms other than phosphate. The first part of this investigation was to isolate from Cape Canaveral soil micro-organisms capable of utilizing phosphite as their phosphorus source under anaerobic conditions. In an attempt to demonstrate this ability, a medium was prepared which contained hypophosphite as the phosphorus source. This was inoculated with soil samples, and growth was subcultured at least four times. To verify that these isolates could use hypophosphite, they were inoculated into defined hypophosphite medium, and samples were removed periodically and killed with formalin. Growth was determined by turbidity measurements and the sample was then filtered. The filtrate was separated by chromatography and the total amounts of hypophosphite, phosphate and phosphate in the filtrate were measured. By this procedure it appeared that the hypophosphite level began decreasing after 14 hr of incubation suggesting utilization of the hypophosphite under anaerobic conditions. The third part of this investigation used labeled (32P) hypophosphite in a defined medium; the cells were then lysed and the metabolic compounds separated by the use of paper chromatography and autoradiograms, demonstrating the presence of 32P in intermediate metabolic compounds. Similar investigations are now being performed with phosphine as the phosphorus source.

  2. Comparative study on the uptake and bioimpact of metal nanoparticles released into environment

    NASA Astrophysics Data System (ADS)

    Andries, Maria; Pricop, Daniela; Grigoras, Marian; Lupu, Nicoleta; Sacarescu, Liviu; Creanga, Dorina; Iacomi, Felicia

    2015-12-01

    Metallic particles of very small size are ubiquitously released in the air, water and soil from various natural and artificial sources - the last ones with enhanced extent since nanotechnology development accelerated exponentially. In this study we focused on the impact of metal nanoparticles in vegetal species of agroindustrial interest namely the maize (Zea mais L.). Laboratory simulation of environmental pollution was carried out by using engineered nanoparticles of two types: iron oxides with magnetic properties and gold nanoparticles supplied in the form of dilutes stable suspensions in the culture medium of maize seedlings. Magnetic nanoparticle (MNPs) preparation was performed by applying chemical route from iron ferric and ferrous precursor salts in alkali reaction medium at relatively high temperature (over 80 °C). Gold nanoparticles (GNPs) synthesis was accomplished from auric hydrochloride acid in alkali reaction medium in similar temperature conditions. In both types of metallic nanoparticles citrate ions were used as coating shell with role of suspension stabilization. Plantlet response was assessed at the level of assimilatory pigment contents in green tissue of seedlings in early ontogenetic stages.

  3. Laser activation of a nutrient medium and antibiotic solutions and its estimation by of bacteria growth dynamics

    NASA Astrophysics Data System (ADS)

    Malov, Alexander N.; Neupokoeva, Anna V.; Kokorina, Lubov A.; Simonova, Elena V.

    2016-11-01

    A laser photomodifacation of nutrient mediums and antibiotics results at the microbiological supervision of bacteria colonies growth are discussed. It is experimentally shown, that on the irradiated media there is a delay of bacterial colonies growth number. Influence of laser radiation on activity of an antibiotic also is experimentally studied. It is revealed, that laser photomodifacation increases antimicrobic activity of a preparation. The mechanism of biological solutions activation is connected with the phenomenon laser nanoclusterization. Parameters of bacteria growth bacteria growth dynamics allow to numerically estimate degree of laser activation of nutrient mediums and pharmaceutical preparations.

  4. 2,2-Diphenyl-1-picrylhydrazyl as a screening tool for recombinant monoterpene biosynthesis.

    PubMed

    Behrendorff, James Byh; Vickers, Claudia E; Chrysanthopoulos, Panagiotis; Nielsen, Lars K

    2013-08-23

    Monoterpenes are a class of natural C10 compounds with a range of potential applications including use as fuel additives, fragrances, and chemical feedstocks. Biosynthesis of monoterpenes in heterologous systems is yet to reach commercially-viable levels, and therefore is the subject of strain engineering and fermentation optimization studies. Detection of monoterpenes typically relies on gas chromatography/mass spectrometry; this represents a significant analytical bottleneck which limits the potential to analyse combinatorial sets of conditions. To address this, we developed a high-throughput method for pre-screening monoterpene biosynthesis. An optimised DPPH assay was developed for detecting monoterpenes from two-phase microbial cultures using dodecane as the extraction solvent. The assay was useful for reproducible qualitative ranking of monoterpene concentrations, and detected standard preparations of myrcene and γ-terpinene dissolved in dodecane at concentrations as low as 10 and 15 μM, respectively, and limonene as low as 200 μM. The assay could not be used quantitatively due to technical difficulties in capturing the initial reaction rate in a multi-well plate and the presence of minor DPPH-reactive contaminants. Initially, limonene biosynthesis in Saccharomyces cerevisiae was tested using two different limonene synthase enzymes and three medium compositions. The assay indicated that limonene biosynthesis was enhanced in a supplemented YP medium and that the Citrus limon limonene synthase (CLLS) was more effective than the Mentha spicata limonene synthase (MSLS). GC-MS analysis revealed that the DPPH assay had correctly identified the best limonene synthase (CLLS) and culture medium (supplemented YP medium). Because only traces of limonene were detected in SD medium, we subsequently identified medium components that improved limonene production and developed a defined medium based on these findings. The best limonene titres obtained were 1.48 ± 0.22 mg limonene per L in supplemented YP medium and 0.9 ± 0.15 mg limonene per L in a pH-adjusted supplemented SD medium. The DPPH assay is useful for detecting biosynthesis of limonene. Although the assay cannot be used quantitatively, it proved successful in ranking limonene production conditions qualitatively and thus is suitable as a first-tier screen. The DPPH assay will likely be applicable in detecting biosynthesis of several other monoterpenes and for screening libraries of monoterpene-producing strains.

  5. 2,2-Diphenyl-1-picrylhydrazyl as a screening tool for recombinant monoterpene biosynthesis

    PubMed Central

    2013-01-01

    Background Monoterpenes are a class of natural C10 compounds with a range of potential applications including use as fuel additives, fragrances, and chemical feedstocks. Biosynthesis of monoterpenes in heterologous systems is yet to reach commercially-viable levels, and therefore is the subject of strain engineering and fermentation optimization studies. Detection of monoterpenes typically relies on gas chromatography/mass spectrometry; this represents a significant analytical bottleneck which limits the potential to analyse combinatorial sets of conditions. To address this, we developed a high-throughput method for pre-screening monoterpene biosynthesis. Results An optimised DPPH assay was developed for detecting monoterpenes from two-phase microbial cultures using dodecane as the extraction solvent. The assay was useful for reproducible qualitative ranking of monoterpene concentrations, and detected standard preparations of myrcene and γ-terpinene dissolved in dodecane at concentrations as low as 10 and 15 μM, respectively, and limonene as low as 200 μM. The assay could not be used quantitatively due to technical difficulties in capturing the initial reaction rate in a multi-well plate and the presence of minor DPPH-reactive contaminants. Initially, limonene biosynthesis in Saccharomyces cerevisiae was tested using two different limonene synthase enzymes and three medium compositions. The assay indicated that limonene biosynthesis was enhanced in a supplemented YP medium and that the Citrus limon limonene synthase (CLLS) was more effective than the Mentha spicata limonene synthase (MSLS). GC-MS analysis revealed that the DPPH assay had correctly identified the best limonene synthase (CLLS) and culture medium (supplemented YP medium). Because only traces of limonene were detected in SD medium, we subsequently identified medium components that improved limonene production and developed a defined medium based on these findings. The best limonene titres obtained were 1.48 ± 0.22 mg limonene per L in supplemented YP medium and 0.9 ± 0.15 mg limonene per L in a pH-adjusted supplemented SD medium. Conclusions The DPPH assay is useful for detecting biosynthesis of limonene. Although the assay cannot be used quantitatively, it proved successful in ranking limonene production conditions qualitatively and thus is suitable as a first-tier screen. The DPPH assay will likely be applicable in detecting biosynthesis of several other monoterpenes and for screening libraries of monoterpene-producing strains. PMID:23968454

  6. Vibrio sp. DSM 14379 pigment production--a competitive advantage in the environment?

    PubMed

    Starič, Nejc; Danevčič, Tjaša; Stopar, David

    2010-10-01

    The ability to produce several antibacterial agents greatly increases the chance of producer's survival. In this study, red-pigmented Vibrio sp. DSM 14379 and Bacillus sp., both isolated from the same sampling volume from estuarine waters of the Northern Adriatic Sea, were grown in a co-culture. The antibacterial activity of the red pigment extract was tested on Bacillus sp. in microtiter plates. The MIC(50) for Bacillus sp. was estimated to be around 10⁻⁵ mg/L. The extract prepared form the nonpigmented mutant of Vibrio sp. had no antibacterial effect. The pigment production of Vibrio sp. was studied under different physicochemical conditions. There was no pigment production at high or low temperatures, high or low salt concentrations in peptone yeast extract (PYE) medium, low glucose concentration in mineral growth medium or high glucose concentration in PYE medium. This indicates that the red pigment production is a luxurious good that Vibrio sp. makes only under favorable conditions. The Malthusian fitness of Bacillus sp. in a co-culture with Vibrio sp. under optimal environmental conditions dropped from 4.0 to -7.6, which corresponds to three orders of magnitude decrease in the number of CFU relative to the monoculture. The nonpigmented mutant of Vibrio sp. in a co-culture with Bacillus sp. had a significant antibacterial activity. This result shows that studying antibacterial properties in isolation (i.e. pigment extract only) may not reveal full antibacterial potential of the bacterial strain. The red pigment is a redundant antibacterial agent of Vibrio sp.

  7. Influenza pandemic preparedness: motivation for protection among small and medium businesses in Australia

    PubMed Central

    Watkins, Rochelle E; Cooke, Feonagh C; Donovan, Robert J; MacIntyre, C Raina; Itzwerth, Ralf; Plant, Aileen J

    2007-01-01

    Background Community-wide preparedness for pandemic influenza is an issue that has featured prominently in the recent news media, and is currently a priority for health authorities in many countries. The small and medium business sector is a major provider of private sector employment in Australia, yet we have little information about the preparedness of this sector for pandemic influenza. This study aimed to investigate the association between individual perceptions and preparedness for pandemic influenza among small and medium business owners and managers. Methods Semi-structured face-to-face interviews were conducted with 201 small and medium business owners or managers in New South Wales and Western Australia. Eligible small or medium businesses were defined as those that had less than 200 employees. Binomial logistic regression analysis was used to identify the predictors of having considered the impact of, having a plan for, and needing help to prepare for pandemic influenza. Results Approximately 6 per cent of participants reported that their business had a plan for pandemic influenza, 39 per cent reported that they had not thought at all about the impact of pandemic influenza on their business, and over 60 per cent stated that they required help to prepare for a pandemic. Beliefs about the severity of pandemic influenza and the ability to respond were significant independent predictors of having a plan for pandemic influenza, and the perception of the risk of pandemic influenza was the most important predictor of both having considered the impact of, and needing help to prepare for a pandemic. Conclusion Our findings suggest that small and medium businesses in Australia are not currently well prepared for pandemic influenza. We found that beliefs about the risk, severity, and the ability to respond effectively to the threat of pandemic influenza are important predictors of preparedness. Campaigns targeting small and medium businesses should emphasise the severity of the consequences to their businesses if a pandemic were to occur, and, at the same time, reassure them that there are effective strategies capable of being implemented by small and medium businesses to deal with a pandemic. PMID:17634112

  8. Influenza pandemic preparedness: motivation for protection among small and medium businesses in Australia.

    PubMed

    Watkins, Rochelle E; Cooke, Feonagh C; Donovan, Robert J; MacIntyre, C Raina; Itzwerth, Ralf; Plant, Aileen J

    2007-07-17

    Community-wide preparedness for pandemic influenza is an issue that has featured prominently in the recent news media, and is currently a priority for health authorities in many countries. The small and medium business sector is a major provider of private sector employment in Australia, yet we have little information about the preparedness of this sector for pandemic influenza. This study aimed to investigate the association between individual perceptions and preparedness for pandemic influenza among small and medium business owners and managers. Semi-structured face-to-face interviews were conducted with 201 small and medium business owners or managers in New South Wales and Western Australia. Eligible small or medium businesses were defined as those that had less than 200 employees. Binomial logistic regression analysis was used to identify the predictors of having considered the impact of, having a plan for, and needing help to prepare for pandemic influenza. Approximately 6 per cent of participants reported that their business had a plan for pandemic influenza, 39 per cent reported that they had not thought at all about the impact of pandemic influenza on their business, and over 60 per cent stated that they required help to prepare for a pandemic. Beliefs about the severity of pandemic influenza and the ability to respond were significant independent predictors of having a plan for pandemic influenza, and the perception of the risk of pandemic influenza was the most important predictor of both having considered the impact of, and needing help to prepare for a pandemic. Our findings suggest that small and medium businesses in Australia are not currently well prepared for pandemic influenza. We found that beliefs about the risk, severity, and the ability to respond effectively to the threat of pandemic influenza are important predictors of preparedness. Campaigns targeting small and medium businesses should emphasise the severity of the consequences to their businesses if a pandemic were to occur, and, at the same time, reassure them that there are effective strategies capable of being implemented by small and medium businesses to deal with a pandemic.

  9. Eco-friendly intracellular biosynthesis of CdS quantum dots without changing Escherichia coli's antibiotic resistance.

    PubMed

    Yan, Zheng-Yu; Du, Qing-Qing; Qian, Jing; Wan, Dong-Yu; Wu, Sheng-Mei

    2017-01-01

    In the paper, a green and efficient biosynthetical technique was reported for preparing cadmium sulfide (CdS) quantum dots, in which Escherichia coli (E. coli) was chosen as a biomatrix. Fluorescence emission spectra and fluorescent microscopic photographs revealed that as-produced CdS quantum dots had an optimum fluorescence emission peak located at 470nm and emitted a blue-green fluorescence under ultraviolet excitation. After extracted from bacterial cells and located the nanocrystals' foci in vivo, the CdS quantum dots showed a uniform size distribution by transmission electron microscope. Through the systematical investigation of the biosynthetic conditions, including culture medium replacement, input time point of cadmium source, working concentrations of raw inorganic ions, and co-cultured time spans of bacteria and metal ions in the bio-manufacture, the results revealed that CdS quantum dots with the strongest fluorescence emission were successfully prepared when E. coli cells were in stationary phase, with the replacement of culture medium and following the incubation with 1.0×10 -3 mol/L cadmium source for 2 days. Results of antimicrobial susceptibility testing indicated that the sensitivities to eight types of antibiotics of E. coli were barely changed before and after CdS quantum dots were prepared in the mild temperature environment, though a slight fall of antibiotic resistance could be observed, suggesting hinted the proposed technique of producing quantum dots is a promising environmentally low-risk protocol. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Degradation and protein release properties of microspheres prepared from biodegradable poly(lactide-co-glycolide) and ABA triblock copolymers: influence of buffer media on polymer erosion and bovine serum albumin release.

    PubMed

    Bittner, B; Witt, C; Mäder, K; Kissel, T

    1999-08-05

    The aim of the present study was to investigate the influence of the chemical insertion of poly(ethylene oxide), PEO, into a poly(lactide-co-glycolide), PLG, backbone on the mechanisms of in vitro degradation and erosion of the polymer. For this purpose microspheres prepared by a modified W/O/W double emulsion technique using ABA triblock copolymers, consisting of PLG A-blocks attached to central PEO B-blocks were compared with microspheres prepared from PLG. Due to their molecular architecture the ABA triblock copolymers differed in their erosion and degradation behavior from PLG. Degradation occurred faster in the ABA polymers by cleavage of ester bonds inside the polymer backbone. Even erosion was shown to start immediately after incubation in different buffer media. By varying pH and ionic strength of the buffer it was found that both mass loss and molecular weight decay were accelerated in alkaline and acidic pH in the case of the ABA triblock copolymers. Although the pH of the medium had a moderate influence on the degradation of PLG, the molecular weight decay was not accompanied by a mass loss during the observation time. In a second set of experiments we prepared bovine serum albumin, BSA, loaded microspheres from both polymers. The release of BSA from ABA microspheres under in vitro conditions parallels the faster swelling and erosion rates. This could be confirmed by electron paramagnetic resonance, EPR, measurements with spin labeled albumin where an influx of buffer medium into the ABA microspheres was already observed within a few minutes. In contrast, PLG microspheres revealed a burst release without any erosion. The current study shows that the environmental conditions affected the degradation and erosion of the pure polymer microspheres in the same way as the release of the model protein. This leads to the conclusion that the more favorable degradation profile of the ABA triblock copolymers was responsible for the improvement of the release profile.

  11. Quantifying the Impact of Food Preparation Skills among College Women

    ERIC Educational Resources Information Center

    Soliah, LuAnn; Walter, Janelle; Antosh, Deeanna

    2006-01-01

    Food preparation practices have changed dramatically in the past fifty years. This study was designed to quantify food preparation knowledge and practices of college women, to assess the reasons why they do not prepare certain foods, and to determine the frequency of eating outside the home. Distinct food preparation ability emerged (high, medium,…

  12. 75 FR 33565 - Notice of Intent To Prepare an Environmental Impact Statement for New Medium- and Heavy-Duty Fuel...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-14

    ...- and Heavy-Duty Fuel Efficiency Improvement Program AGENCY: National Highway Traffic Safety... efficiency improvement program for commercial medium- and heavy-duty on-highway vehicles and work trucks... efficiency standards starting with model year (MY) 2016 commercial medium- and heavy-duty on-highway vehicles...

  13. The end-stage renal disease industry and exit strategies for nephrologists.

    PubMed

    Sullivan, John D

    2006-01-01

    The purpose of this presentation is to identify exit strategies for nephrologists under changing conditions in the dialysis market. The end-stage renal disease service provider market continues to be highly receptive to consolidation. Taking advantage of large economies of scale, large for-profit dialysis chains have surpassed independent operators in both number of clinics and total patients. With relatively low barriers to entry, new smaller clinics continue to open, serving a niche outside the larger chains. Additional competition comes in the form of medium players funded by venture capitalists with the added pressure of rapid growth and financial return. To ensure market power in both dialysis products and managed care negotiation leverage, medium and large service providers will continue to seek out attractive acquisition targets. For nephrologists to capitalize on investment, clinic and business preparation will continue to be the driving force for these divestitures.

  14. Concentration, physical state, and purity of bacterial endotoxin affect its detoxification by ionizing radiation

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

    Csako, G.; Tsai, C.M.; Hochstein, H.D.

    Increasing concentrations of a highly purified bacterial lipopolysaccharide preparation, the U.S. Reference Standard Endotoxin, were exposed to increasing doses of ionizing radiation from a 60Co source. At identical radiation doses both the structural change and Limulus amebocyte lysate (LAL) reactivity were progressively smaller with increasing concentrations of the lipopolysaccharide in an aqueous medium. Under the experimental conditions used, there was a linear relationship between the endotoxin concentration and radiation dose for the structural changes. In contrast to endotoxin in aqueous medium, endotoxin irradiated in its dry state showed no decrease in LAL reactivity and rabbit pyrogenicity. Endotoxin exposed to radiationmore » in water in the presence of albumin showed a much smaller decrease in LAL and pyrogenic activities than expected. The results show that the concentration, physical state, and purity of endotoxin influence its structural and functional alteration by ionizing radiation.« less

  15. Preparation and characterization of a thermoresponsive gigaporous medium for high-speed protein chromatography.

    PubMed

    Qu, Jian-Bo; Chen, Yan-Li; Huan, Guan-Sheng; Zhou, Wei-Qing; Liu, Jian-Guo; Zhu, Hu; Zhang, Xiao-Yun

    2015-01-01

    A high-speed thermoresponsive medium was developed by grafting poly(N-isopropylacrylamide-co-butyl methacrylate) (P(NIPAM-co-BMA)) brushes onto gigaporous polystyrene (PS) microspheres via surface-initiated atom transfer radical polymerization (ATRP) technique, which has strong mechanical strength, good chemical stability and high mass transfer rate for biomacromolecules. The gigaporous structure, surface chemical composition, static protein adsorption, and thermoresponsive chromatographic properties of prepared medium (PS-P(NIPAM-co-BMA)) were characterized in detail. Results showed that the PS microspheres were successfully grafted with P(NIPAM-co-BMA) brushes and that the gigaporous structure was robustly maintained. After grafting, the nonspecific adsorption of proteins on PS microspheres was greatly reduced. A column packed with PS-P(NIPAM-co-BMA) exhibited low backpressure and significant thermo-responsibility. By simply changing the column temperature, it was able to separate three model proteins at the mobile phase velocity up to 2167 cm h(-1). In conclusion, the thermoresponsive polymer brushes grafted gigaporous PS microspheres prepared by ATRP are very promising in 'green' high-speed preparative protein chromatography. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Anaerobic digestion of citrus waste using two-stage membrane bioreactor

    NASA Astrophysics Data System (ADS)

    Millati, Ria; Lukitawesa; Dwi Permanasari, Ervina; Wulan Sari, Kartika; Nur Cahyanto, Muhammad; Niklasson, Claes; Taherzadeh, Mohammad J.

    2018-03-01

    Anaerobic digestion is a promising method to treat citrus waste. However, the presence of limonene in citrus waste inhibits anaerobic digestion process. Limonene is an antimicrobial compound and could inhibit methane forming bacteria that takes a longer time to recover than the injured acid forming bacteria. Hence, volatile fatty acids will be accumulated and methane production will be decreased. One way to solve this problem is by conducting anaerobic digestion process into two stages. The first step is aimed for hydrolysis, acidogenesis, and acetogenesis reactions and the second stage is aimed for methanogenesis reaction. The separation of the system would further allow each stage in their optimum conditions making the process more stable. In this research, anaerobic digestion was carried out in batch operations using 120 ml-glass bottle bioreactors in 2 stages. The first stage was performed in free-cells bioreactor, whereas the second stage was performed in both bioreactor of free cells and membrane bioreactor. In the first stage, the reactor was set into ‘anaerobic’ and ‘semi-aerobic’ conditions to examine the effect of oxygen on facultative anaerobic bacteria in acid production. In the second stage, the protection of membrane towards the cells against limonene was tested. For the first stage, the basal medium was prepared with 1.5 g VS of inoculum and 4.5 g VS of citrus waste. The digestion process was carried out at 55°C for four days. For the second stage, the membrane bioreactor was prepared with 3 g of cells that were encased and sealed in a 3×6 cm2 polyvinylidene fluoride membrane. The medium contained 40 ml basal medium and 10 ml liquid from the first stage. The bioreactors were incubated at 55°C for 2 days under anaerobic condition. The results from the first stage showed that the maximum total sugar under ‘anaerobic’ and ‘semi-aerobic’ conditions was 294.3 g/l and 244.7 g/l, respectively. The corresponding values for total volatile fatty acids were 3.8 g/l and 2.9 g/l, respectively. Methane production of citrus waste taken from the first stage under ‘anaerobic’ condition in membrane and free-cells bioreactors was 11.2 Nml and 7.2 Nml, respectively. Whereas, methane production of citrus waste taken from the first stage under ‘semi-aerobic’ condition in membrane and free-cells bioreactors was 8.8 Nml and 5.7 Nml, respectively. It can be seen from the results of the first stage that volatile fatty acids from ‘anaerobic’ condition was higher than that of ‘semi-aerobic’ condition. The absence of oxygen provides the optimal condition for growth and metabolism of facultative and obligatorily anaerobic bacteria in the first stage. Furthermore, polyvinylidene fluoride membrane was able to protect the cells from antimicrobial compounds.

  17. Polyacrylamide medium for the electrophoretic separation of biomolecules

    DOEpatents

    Madabhushi, Ramakrishna S.; Gammon, Stuart A.

    2003-11-11

    A polyacryalmide medium for the electrophoretic separation of biomolecules. The polyacryalmide medium comprises high molecular weight polyacrylamides (PAAm) having a viscosity average molecular weight (M.sub.v) of about 675-725 kDa were synthesized by conventional red-ox polymerization technique. Using this separation medium, capillary electrophoresis of BigDye DNA sequencing standard was performed. A single base resolution of .about.725 bases was achieved in .about.60 minute in a non-covalently coated capillary of 50 .mu.m i.d., 40 cm effective length, and a filed of 160 V/cm at 40.degree. C. The resolution achieved with this formulation to separate DNA under identical conditions is much superior (725 bases vs. 625 bases) and faster (60 min. vs. 75 min.) to the commercially available PAAm, such as supplied by Amersham. The formulation method employed here to synthesize PAAm is straight-forward, simple and does not require cumbersome methods such as emulsion polymerizaiton in order to achieve very high molecular weights. Also, the formulation here does not require separation of PAAm from the reaction mixture prior to reconstituting the polymer to a final concentration. Furthermore, the formulation here is prepared from a single average mol. wt. PAAm as opposed to the mixture of two different average mo. wt. PAAm previously required to achieve high resolution.

  18. [Nutritional value of daily diets prepared in several regions of the country. Part VI. Value of retinol, beta-carotene and vitamin E].

    PubMed

    Nadolna, I; Okolska, G; Kunachowicz, H

    1992-01-01

    Determinations of retinol, beta-carotene and vitamin E were carried out in daily diets of manual and mental workers with medium incomes. The diets were prepared on the basis of the analysis of the Central Statistical Office in 1986 under laboratory conditions in five regions of Poland (Warszawa, Lublin, Olsztyn, Poznań, Wrocław). It was found that these diets contained respectively: vitamin A (retinol and beta-carotene) 844 and 859 micrograms, retinol 466 and 496 micrograms, beta-carotene 2267 and 2167 micrograms and vitamin E 4.82 and 5.12 mg per day. In relation to the realization of daily recommended dietary allowances by these diets, for vitamin A were met respectively 102 and 104% and for vitamin E 58 and 61%. There were considerable differences in the content of beta-carotene and vitamin E between diets prepared in all five regions of Poland.

  19. Insulin-loaded W/O/W multiple emulsions: comparison of the performances of systems prepared with medium-chain-triglycerides and fish oil.

    PubMed

    Cournarie, Fabienne; Savelli, Marie-Pierre; Rosilio, Véronique; Bretez, Françoise; Vauthier, Christine; Grossiord, Jean-Louis; Seiller, Monique

    2004-11-01

    Insulin-loaded W/O/W multiple emulsions (ME) composed of medium-chain triglycerides have been shown to decrease the blood glucose level after oral administration to diabetic rats. Fish oil (very long-chain triglycerides) could be an alternative to medium-chain triglycerides because its chronic consumption has beneficial therapeutic effects. The aim of this work was twofold: to obtain stable fish oil containing ME, based on a formulation optimized in a previous work with low medium-chain triglycerides content, and to compare their characteristics to those of ME composed of medium-chain triglycerides. Due to the higher viscosity and surface tension of fish oil compared to medium-chain triglycerides, preparation of ME appeared difficult to achieve. However, a stable unloaded-ME with low fish oil content was formed, by adapting the emulsification process. The characteristics of unloaded fish oil ME were almost similar to those of medium-chain triglycerides ME. In contrast to medium-chain triglycerides ME, the introduction of insulin did not improve the elasticity and consequently the characteristics and stability of fish oil ME. Nevertheless, the insulin-loaded fish oil containing ME was shown to be stable for 6 weeks at 4 degrees C.

  20. CD-ROM preparation: An overview and guide

    NASA Technical Reports Server (NTRS)

    Daniel, Ralph E.; Jeschke, Mark W.; Schroer, James A.

    1995-01-01

    A primer on the options and procedures involved in producing CD-ROM products in a small to medium sized business operation is presented in language that persons with a minimal technical background can easily understand. The capabilities, limitations, and standards of CD-ROM technology are surveyed. Emphasis is placed on CD-ROM production, especially upon design, data conversion to an electronic medium, data file preparation, the use of vendors, and the steps for in-house production of CD-ROM products.

  1. Immobilisation increases yeast cells' resistance to dehydration-rehydration treatment.

    PubMed

    Borovikova, Diana; Rozenfelde, Linda; Pavlovska, Ilona; Rapoport, Alexander

    2014-08-20

    This study was performed with the goal of revealing if the dehydration procedure used in our new immobilisation method noticeably decreases the viability of yeast cells in immobilised preparations. Various yeasts were used in this research: Saccharomyces cerevisiae cells that were rather sensitive to dehydration and had been aerobically grown in an ethanol-containing medium, a recombinant strain of S. cerevisiae grown in aerobic conditions which were completely non-resistant to dehydration and an anaerobically grown bakers' yeast strain S. cerevisiae, as well as a fairly resistant Pichia pastoris strain. Experiments performed showed that immobilisation of all these strains essentially increased their resistance to a dehydration-rehydration treatment. The increase of cells' viability (compared with control cells dehydrated in similar conditions) was from 30 to 60%. It is concluded that a new immobilisation method, which includes a dehydration stage, does not lead to an essential loss of yeast cell viability. Correspondingly, there is no risk of losing the biotechnological activities of immobilised preparations. The possibility of producing dry, active yeast preparations is shown, for those strains that are very sensitive to dehydration and which can be used in biotechnology in an immobilised form. Finally, the immobilisation approach can be used for the development of efficient methods for the storage of recombinant yeast strains. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Water-Soluble Nonconjugated Polymer Nanoparticles with Strong Fluorescence Emission for Selective and Sensitive Detection of Nitro-Explosive Picric Acid in Aqueous Medium.

    PubMed

    Liu, Shi Gang; Luo, Dan; Li, Na; Zhang, Wei; Lei, Jing Lei; Li, Nian Bing; Luo, Hong Qun

    2016-08-24

    Water-soluble nonconjugated polymer nanoparticles (PNPs) with strong fluorescence emission were prepared from hyperbranched poly(ethylenimine) (PEI) and d-glucose via Schiff base reaction and self-assembly in aqueous phase. Preparation of the PEI-d-glucose (PEI-G) PNPs was facile (one-pot reaction) and environmentally friendly under mild conditions. Also, PEI-G PNPs showed a high fluorescence quantum yield in aqueous solution, and the fluorescence properties (such as concentration- and solvent-dependent fluorescence) and origin of intrinsic fluorescence were investigated and discussed. PEI-G PNPs were then used to develop a fluorescent probe for fast, selective, and sensitive detection of nitro-explosive picric acid (PA) in aqueous medium, because the fluorescence can be easily quenched by PA whereas other nitro-explosives and structurally similar compounds only caused negligible quenching. A wide linear range (0.05-70 μM) and a low detection limit (26 nM) were obtained. The fluorescence quenching mechanism was carefully explored, and it was due to a combined effect of electron transfer, resonance energy transfer, and inner filter effect between PA and PEI-G PNPs, which resulted in good selectivity and sensitivity for PA. Finally, the developed sensor was successfully applied to detection of PA in environmental water samples.

  3. [Polyelectrolyte microcapsules as systems for delivery of biologically active substances].

    PubMed

    Borodina, T N; Rumsh, L D; Kunizhev, S M; Sukhorukov, G B; Vorozhtsov, G N; Fel'dman, B M; Markvicheva, E A

    2007-01-01

    Novel biodegradable microcapsules for delivery of biologically active substances (BAS) were prepared by layer-by-layer (LbL) adsorption of oppositely charged polyelectrolytes, namely sodium alginate (Alg) and poly-L-lysine (PLL). To immobilize these BAS, porous spherical CaCO3 microparticles were used as templates. The templates (cores) were coated with several layers of oppositely charged polyelectrolytes forming shell on a core surface. The core-shell microparticles were converted into hollow microcapsules by a core dissolution after an EDTA treatment. Mild conditions for microcapsule fabrication allow to perform an entrapment of various biomolecules while keeping their bioactivity. Biocompatibility and biodegradable capability of the polyelectrolytes give a possibility to use the microcapsules as the target delivery systems. Chymotrypsin (Chym) entrapped into the microcapsules was used as a model enzyme. The immobilized enzyme was found to keep about 86% of the activity compared to a native Chym. The obtained microcapsules were stable at an acidic medium while they could be easily decomposed by trypsin treatment at an slightly alkaline medium. Chym was shown to be active after being released from the microcapsules decomposed by trypsin treatment. Thus, the microcapsules prepared by the LbL - technique can be used for the development of new type of BAS delivery systems in humans and animals.

  4. Depth enhancement of ion sensitized data

    DOEpatents

    Lamartine, Bruce C.

    2001-01-01

    A process of fabricating a durable data storage medium is disclosed, the durable data storage medium capable of storing, digital or alphanumeric characters as well as graphical shapes or characters. Additionally, a durable data storage medium including a substrate having etched characters therein is disclosed, the substrate characterized as containing detectable residual amounts of ions used in the preparation process.

  5. [Microfungicid--a preparation based on trichoderma viride for plant diseases control].

    PubMed

    Kolombet, L V; Zhigletsova, S K; Derbyshev, V V; Ezhov, D V; Kosareva, N I; Bystrova, E V

    2001-01-01

    A technology was designed for manufacturing a preparation based on Trichoderma viride Pers ex S.F. Gray that strongly suppresses the development of causative agents of certain plant diseases and displays a growth-stimulating activity. Cultivation of the strain in a liquid medium for 18-24 h produced up to 60 g dry biomass per liter nutrient medium. A marketable form created in this work conserves the activity of the mycelial preparation for six months. The preparation is compatible with insecticides (carbofos, vismetrin, talstar, and applaud) and certain fungicides (such as baitan). Tests performed with the liquid form of Mycofungicid (seeds were treated with this preparation at a dose of 20-30 g per metric ton before sowing) showed its high efficiency in protecting cereal crops from plant pathogens. The incidence of plant diseases decreased by 65%, and crop yields increased by 15-20%.

  6. Calibration of redox potential in sperm wash media and evaluation of oxidation-reduction potential values in various assisted reproductive technology culture media using MiOXSYS system.

    PubMed

    Panner Selvam, M K; Henkel, R; Sharma, R; Agarwal, A

    2018-03-01

    Oxidation-reduction potential describes the balance between the oxidants and antioxidants in fluids including semen. Various artificial culture media are used in andrology and IVF laboratories for sperm preparation and to support the development of fertilized oocytes under in vitro conditions. The composition and conditions of these media are vital for optimal functioning of the gametes. Currently, there are no data on the status of redox potential of sperm processing and assisted reproduction media. The purpose of this study was to compare the oxidation-reduction potential values of the different media and to calibrate the oxidation-reduction potential values of the sperm wash medium using oxidative stress inducer cumene hydroperoxide and antioxidant ascorbic acid. Redox potential was measured in 10 different media ranging from sperm wash media, freezing media and assisted reproductive technology one-step medium to sequential media. Oxidation-reduction potential values of the sequential culture medium and one-step culture medium were lower and significantly different (p < 0.05) from the sperm wash media. Calibration of the sperm wash media using the oxidant cumene hydroperoxide and antioxidant ascorbic acid demonstrated that oxidation-reduction potential and the concentration of oxidant or antioxidant are logarithmically dependent. This study highlights the importance of calibrating the oxidation-reduction potential levels of the sperm wash media in order to utilize it as a reference value to identify the physiological range of oxidation-reduction potential that does not have any adverse effect on normal physiological sperm function. © 2017 American Society of Andrology and European Academy of Andrology.

  7. Buffer-free therapeutic antibody preparations provide a viable alternative to conventionally buffered solutions: from protein buffer capacity prediction to bioprocess applications.

    PubMed

    Bahrenburg, Sven; Karow, Anne R; Garidel, Patrick

    2015-04-01

    Protein therapeutics, including monoclonal antibodies (mAbs), have significant buffering capacity, particularly at concentrations>50 mg/mL. This report addresses pH-related issues critical to adoption of self-buffered monoclonal antibody formulations. We evaluated solution conditions with protein concentrations ranging from 50 to 250 mg/mL. Samples were both buffer-free and conventionally buffered with citrate. Samples were non-isotonic or adjusted for isotonicity with NaCl or trehalose. Studies included accelerated temperature stability tests, shaking stability studies, and pH changes in infusion media as protein concentrate is added. We present averaged buffering slopes of capacity that can be applied to any mAb and present a general method for calculating buffering capacity of buffer-free, highly concentrated antibody liquid formulations. In temperature stability tests, neither buffer-free nor conventionally buffered solution conditions showed significant pH changes. Conventionally buffered solutions showed significantly higher opalescence than buffer-free ones. In general, buffer-free solution conditions showed less aggregation than conventionally buffered solutions. Shaking stability tests showed no differences between buffer-free and conventionally buffered solutions. "In-use" preparation experiments showed that pH in infusion bag medium can rapidly approximate that of self-buffered protein concentrate as concentrate is added. In summary, the buffer capacity of proteins can be predicted and buffer-free therapeutic antibody preparations provide a viable alternative to conventionally buffered solutions. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Preparing sternorrhynchous insects (Insecta: Hemiptera: Sternorrhyncha) for microscope examination: Hoyer’s mounting medium

    USDA-ARS?s Scientific Manuscript database

    Proper identification of aphids, scale insects, psyllids, and whitefles (Hemiptera: Sternorrhyncha) require preparation of the specimen on a microscope slide. This training video provides visual instruction on how to prepare sternorrhynchous specimens on microscope slides for examination and identi...

  9. Notochordal cell conditioned medium stimulates mesenchymal stem cell differentiation toward a young nucleus pulposus phenotype

    PubMed Central

    2010-01-01

    Introduction Mesenchymal stem cells (MSCs) offer promise for intervertebral disc (IVD) repair and regeneration because they are easily isolated and expanded, and can differentiate into several mesenchymal tissues. Notochordal (NC) cells contribute to IVD development, incorporate into the nucleus pulposus (NP), and stimulate mature disc cells. However, there have been no studies investigating the effects of NC cells on adult stem cell differentiation. The premise of this study is that IVD regeneration is more similar to IVD development than to IVD maintenance, and we hypothesize that soluble factors from NC cells differentiate MSCs to a phenotype characteristic of nucleus pulposus (NP) cells during development. The eventual clinical goal would be to isolate or chemically/recombinantly produce the active agent to induce the therapeutic effects, and to use it as either an injectable therapy for early intervention on disc disease, or in developing appropriately pre-differentiated MSC cells in a tissue engineered NP construct. Methods Human MSCs from bone marrow were expanded and pelleted to form high-density cultures. MSC pellets were exposed to either control medium (CM), chondrogenic medium (CM with dexamethasone and transforming growth factor, (TGF)-β3) or notochordal cell conditioned medium (NCCM). NCCM was prepared from NC cells maintained in serum free medium for four days. After seven days culture, MSC pellets were analyzed for appearance, biochemical composition (glycosaminoglycans and DNA), and gene expression profile (sox-9, collagen types-II and III, laminin-β1 and TIMP1(tissue inhibitor of metalloproteinases-1)). Results Significantly higher glycosaminoglycan accumulation was seen in NCCM treated pellets than in CM or TGFβ groups. With NCCM treatment, increased gene expression of collagen III, and a trend of increasing expression of laminin-β1 and decreased expression of sox-9 and collagen II relative to the TGFβ group was observed. Conclusions Together, results suggest NCCM stimulates mesenchymal stem cell differentiation toward a potentially NP-like phenotype with some characteristics of the developing IVD. PMID:20565707

  10. Comparison of mounting methods for the evaluation of fibers by phase contrast microscopy.

    PubMed

    Lee, Eun Gyung; Pang, Thomas W S; Nelson, John; Andrew, Mike; Harper, Martin

    2011-07-01

    The objectives of this study were to evaluate mounting methods for fiber examination of air sample filters by phase contrast microscopy (PCM) and to evaluate differences in fiber counts that might be due to fiber movement. Acetone/triacetin (AT) with various amounts of triacetin and acetone/Euparal (AE) where the mounting medium was placed between the cleared filter wedge and the coverslip were tested as a function of time. Field sample slides collected from a taconite iron-ore processing mill, a tremolitic talc-ore processing mill, and from around a crusher in a meta-basalt stone quarry were prepared with relocatable coverslips to revisit the same field areas on the slides. For each slide, three or four field areas were randomly selected and pictures were taken every 2 weeks to determine any sign of fiber movement over time. For 11 AT slides (named as AT-3.5) prepared with 3.5 μl of the mounting medium according to the NIOSH 7400 method, no fiber movements were detected over 59 weeks. On the other hand, AT slides prepared with larger quantities (10, 15, and 20 μl) of the mounting medium (named as AT-10) and AE slides prepared with ∼10 μl mounting medium showed fiber movement from the eighth day at the earliest. Fiber movement began earlier for the slides mounted with excess triacetin than for those mounted with Euparal. The sample slide storage method, either vertically or horizontally, did not seem to accelerate fiber movement. Additionally, two other modified methods, dimethylformamide solution/Euparal (mDE) and dimethylformamide solution/triacetin (mDT), were also prepared where the mounting medium was placed between the cleared filter wedge and the glass slide. The findings of fiber movements were similar; when 3.5 μl of triacetin was used for the mDT slides, fiber movements were not detected, while fibers on slides prepared with 10 μl triacetin (mDT-10) moved around. No fiber movements were observed for the mDE slides at any time during 59 weeks. Once fiber movement started, fibers moved over distances measured from 4 μm and up to >1000 μm within 22 weeks. However, since then, no further fiber movements have been observed in any field sample slides. Additional sample slides, two Amosite and two chrysotile, were prepared from Union for International Cancer Control (UICC) samples using the AT method with 5 μl triacetin mounting medium. Fiber movements were also observed in these samples; chrysotile fibers began to migrate in 3 weeks, while Amosite fiber movement started after 3 months. Although fiber movement was observed for the AT-10, AE, and mDT-10 sample slides, fiber counts were not significantly different from AT-3.5 and mDE samples that exhibited no fiber movement. Although fiber counts would not be significantly changed by fiber movement, the type and amount of mounting medium for sample slide preparation remains critical for issues such as quality assurance and training of analysts by revisiting the same fibers.

  11. [Biosynthesis of the bioprotectant ectoin by aerobic methylotrophic bacteria from methanol].

    PubMed

    Doronina, N V; Ezhov, V A; Beschastnyĭ, A P; Trotsenko, Iu A

    2010-01-01

    It is shown that neutrophilic methylobacteria Methylophaga thalassica and M. marina have higher rates of growth and ectoin accumulation compared to the haloalkaliphilic species M. alcalica and M. natronia and methanotrophs Methylomicrobium alcaliphilum and M. kenyense. The conditions of M. thalassica cultivation in methanol-containing medium were optimized. The yield of this process reached 60 g/l of absolutely dry biomass containing 15-19% (9-11 g/l) ectoine. The scheme of ectoin isolation from the biomass by extraction and subsequent purification, which allowed obtaining preparations of different degree of purity, was developed.

  12. Preparation, Imaging, and Quantification of Bacterial Surface Motility Assays

    PubMed Central

    Morales-Soto, Nydia; Anyan, Morgen E.; Mattingly, Anne E.; Madukoma, Chinedu S.; Harvey, Cameron W.; Alber, Mark; Déziel, Eric; Kearns, Daniel B.; Shrout, Joshua D.

    2015-01-01

    Bacterial surface motility, such as swarming, is commonly examined in the laboratory using plate assays that necessitate specific concentrations of agar and sometimes inclusion of specific nutrients in the growth medium. The preparation of such explicit media and surface growth conditions serves to provide the favorable conditions that allow not just bacterial growth but coordinated motility of bacteria over these surfaces within thin liquid films. Reproducibility of swarm plate and other surface motility plate assays can be a major challenge. Especially for more “temperate swarmers” that exhibit motility only within agar ranges of 0.4%-0.8% (wt/vol), minor changes in protocol or laboratory environment can greatly influence swarm assay results. “Wettability”, or water content at the liquid-solid-air interface of these plate assays, is often a key variable to be controlled. An additional challenge in assessing swarming is how to quantify observed differences between any two (or more) experiments. Here we detail a versatile two-phase protocol to prepare and image swarm assays. We include guidelines to circumvent the challenges commonly associated with swarm assay media preparation and quantification of data from these assays. We specifically demonstrate our method using bacteria that express fluorescent or bioluminescent genetic reporters like green fluorescent protein (GFP), luciferase (lux operon), or cellular stains to enable time-lapse optical imaging. We further demonstrate the ability of our method to track competing swarming species in the same experiment. PMID:25938934

  13. Control of Drug Dissolution Rate from Film Dosage Forms Containing Valsartan.

    PubMed

    Murata, Yoshifumi; Kofuji, Kyoko; Maida, Chieko

    2016-01-01

    Film dosage forms (FDs) containing valsartan (VST), a popular antihypertensive drug, were prepared using a casting method with sodium alginate and other polysaccharides as the film base. Drug dissolution profiles of the FDs were investigated in limited medium. The FDs were 170-200 μm thick and were easy to handle. All FDs immediately swelled and disintegrated in the medium. About 23% of the VST incorporated into the FD prepared with 1.5% sodium alginate dissolved at 5 min. The initial dissolution rate of VST increased upon the addition of chitosan to the film base; this effect was not observed in the case of chitin. On the other hand, the rate apparently decreased upon modification with alginic acid. In addition, the solubility of VST in the dissolution medium was changed by the addition of chitosan or alginic acid. FDs prepared with polysaccharides are useful for simplifying the administration of drugs to patients, and the drug dissolution rate from FDs can be controlled by modification.

  14. Differences in the ribosomes prepared from lactating and non-lactating bovine mammary gland

    PubMed Central

    Herrington, M. D.; Hawtrey, A. O.

    1971-01-01

    1. Ribosomes prepared from bovine lactating mammary gland are able to synthesize protein, whereas similar preparations from non-lactating glands are not. Washing the ribosome suspensions through a medium containing 0.5m-ammonium chloride enhanced their ability to incorporate phenylalanine into polyphenylalanine. 2. Ribosomes isolated from non-lactating bovine mammary gland, in contrast with those from rat liver and lactating mammary gland, contained significant amounts of extraneous nucleases. These enzymes could be removed by washing with a medium A buffer containing 0.5m-ammonium chloride. 3. Only those ribosomes from functionally active tissues were able to bind polyuridylic acid and phenylalanyl-tRNA. PMID:5165653

  15. The Dynamics of a Five-level (Double Λ)-type Atom Interacting with Two-mode Field in a Cross Kerr-like Medium

    NASA Astrophysics Data System (ADS)

    Obada, A.-S. F.; Ahmed, M. M. A.; Farouk, Ahmed M.

    2018-04-01

    In this paper, we propose a new transition scheme (Double Λ) for the interaction between a five-level atom and an electromagnetic field and study its dynamics in the presence of a cross Kerr-like medium in the exact-resonance case. The wave function is derived when the atom is initially prepared in its upper most state, and the field is initially prepared in the coherent state. We studied the atomic population inversion, the coherence degree by studying the second-order correlation function, Cauchy-Schwartz inequality (CSI) and the relation with P-function. Finally, we investigate the effect of Kerr-like medium on the evolution of Husimi Q-function of the considered system.

  16. Dispersion Polymerization of Polystyrene Particles Using Alcohol as Reaction Medium

    NASA Astrophysics Data System (ADS)

    Cho, Young-Sang; Shin, Cheol Hwan; Han, Sujin

    2016-02-01

    In this study, monodisperse polystyrene nanospheres were prepared by dispersion polymerization using alcohol as reaction medium to prepare colloidal clusters of the latex beads. Polyvinylpyrrolidone (PVP) and 2-(methacryloyloxy)ethyltrimethylammonium chloride (MTC) were used as dispersion stabilizer and comonomer, respectively. The particle size could be controlled by adjusting the reactant compositions such as the amount of stabilizer, comonomer, and water in the reactant mixture. The size and monodispersity of the polymeric particles could be also controlled by changing the reaction medium with different alcohols other than ethanol or adjusting the polymerization temperature. The synthesized particles could be self-organized inside water-in-oil emulsion droplets by evaporation-driven self-assembly to produce colloidal clusters of the polymeric nanospheres.

  17. Prospect of stem cell conditioned medium in regenerative medicine.

    PubMed

    Pawitan, Jeanne Adiwinata

    2014-01-01

    Stem cell-derived conditioned medium has a promising prospect to be produced as pharmaceuticals for regenerative medicine. To investigate various methods to obtain stem cell-derived conditioned medium (CM) to get an insight into their prospect of application in various diseases. Systematic review using keywords "stem cell" and "conditioned medium" or "secretome" and "therapy." Data concerning treated conditions/diseases, type of cell that was cultured, medium and supplements to culture the cells, culture condition, CM processing, growth factors and other secretions that were analyzed, method of application, and outcome were noted, grouped, tabulated, and analyzed. Most of CM using studies showed good results. However, the various CM, even when they were derived from the same kind of cells, were produced by different condition, that is, from different passage, culture medium, and culture condition. The growth factor yields of the various types of cells were available in some studies, and the cell number that was needed to produce CM for one application could be computed. Various stem cell-derived conditioned media were tested on various diseases and mostly showed good results. However, standardized methods of production and validations of their use need to be conducted.

  18. Improved chemically defined basal medium (CMRL-1969) for primary monkey kidney and human diploid cells.

    PubMed

    Healy, G M; Teleki, S; von Seefried, A; Walton, M J; Macmorine, H G

    1971-01-01

    An improved tissue culture basal medium, CMRL-1969, supplemented with serum, has been evaluated by measuring the growth responses of primary cultures of trypsin-dispersed monkey kidney cells (PMKC) and of an established culture of a human diploid cell strain (HDCS). Medium H597, an early modification of medium 199 which has been used successfully in the preparation of poliomyelitis vaccine for 15 years, was used for comparison. In addition, parallel testing was done with Basal Medium Eagle (BME) widely used for the growth of HDCS. The improvements in basal medium CMRL-1969 are attributed to changes in amino acid concentrations, in vitamin composition, and, in particular, to enhanced buffering capacity. The latter has been achieved by the use of free-base amino acids and by increasing the dibasic sodium phosphate. The new medium has already been used to advantage for the production of polioviruses in PMKC where equivalent titers were obtained from cultures initiated with 70% of the number of cells required with earlier media. The population-doubling time was reduced in this system. Also, with small inocula of HDCS, the time required to obtain maximum cell yield was shorter with CMRL-1969 than with BME. Both media were supplemented with 10% calf serum. Maximum cell yields after repeated subcultivation in the new basal medium were greatly increased and the stability of the strain, as shown by chromosomal analysis, was not affected. Basal medium CMRL-1969 can be prepared easily in liquid or powdered form.

  19. Preparation of DNA from cytological material: effects of fixation, staining, and mounting medium on DNA yield and quality.

    PubMed

    Dejmek, Annika; Zendehrokh, Nooreldin; Tomaszewska, Malgorzata; Edsjö, Anders

    2013-07-01

    Personalized oncology requires molecular analysis of tumor cells. Several studies have demonstrated that cytological material is suitable for DNA analysis, but to the authors' knowledge there are no systematic studies comparing how the yield and quality of extracted DNA is affected by the various techniques used for the preparation of cytological material. DNA yield and quality were compared using cultured human lung cancer cells subjected to different preparation techniques used in routine cytology, including fixation, mounting medium, and staining. The results were compared with the outcome of epidermal growth factor receptor (EGFR) genotyping of 66 clinical cytological samples using the same DNA preparation protocol. All tested protocol combinations resulted in fragment lengths of at least 388 base pairs. The mounting agent EcoMount resulted in higher yields than traditional xylene-based medium. Spray and ethanol fixation resulted in both a higher yield and better DNA quality than air drying. In liquid-based cytology (LBC) methods, CytoLyt solution resulted in a 5-fold higher yield than CytoRich Red. Papanicolaou staining provided twice the yield of hematoxylin and eosin staining in both liquid-based preparations. Genotyping outcome and quality control values from the clinical EGFR genotyping demonstrated a sufficient amount and amplifiability of DNA in both spray-fixed and air-dried cytological samples. Reliable clinical genotyping can be performed using all tested methods. However, in the cell line experiments, spray- or ethanol-fixed, Papanicolaou-stained slides provided the best results in terms of yield and fragment length. In LBC, the DNA recovery efficiency of the preserving medium may differ considerably, which should be taken into consideration when introducing LBC. Cancer (Cancer Cytopathol) 2013;121:344-353. © 2013 American Cancer Society. © 2013 American Cancer Society.

  20. Biosynthesis of Ergothioneine from Endogenous Hercynine in Mycobacterium smegmatis

    PubMed Central

    Genghof, Dorothy S.; Van Damme, Olga

    1968-01-01

    Ergothioneine was synthesized and accumulated in growing cultures of Mycobacterium smegmatis when the medium was adequately supplied with sulfur. In a low sulfur medium, the accumulation was sharply limited although growth of the organism was apparently normal. Synthesis of hercynine, the precursor of ergothioneine, was unaffected by low sulfur levels and was markedly increased by addition of l-histidine, the precursor of hercynine. Resting-cell pellicle experiments, performed with cells grown on the low sulfur high histidine medium, showed that ergothioniene was synthesized from endogenous hercynine, when cysteine or compounds readily converted to cysteine (such as cystine, lanthionine, cystathionine, and thiazolidine carboxylic acid) were added. Homocysteine and djenkolic acid allowed for minimal synthesis of betaine, whereas methionine, S-methylcysteine, sodium sulfate, and sodium thiosulfate were unable to donate sulfur for ergothioniene synthesis under the experimental conditions employed. Addition of cysteine to a resting pellicle preparation caused the formation of 100 to 200 μg of ergothioneine per g of dry cells in 2.5 to 3 hr. A modified procedure for isolating ergothioneine and hercynine, employing a 75% ethyl alcohol extraction of wet organisms, followed by a single alumina column separation of the compounds, is described. PMID:5644441

  1. Process for the production of liquid hydrocarbons

    DOEpatents

    Bhatt, Bharat Lajjaram; Engel, Dirk Coenraad; Heydorn, Edward Clyde; Senden, Matthijis Maria Gerardus

    2006-06-27

    The present invention concerns a process for the preparation of liquid hydrocarbons which process comprises contacting synthesis gas with a slurry of solid catalyst particles and a liquid in a reactor vessel by introducing the synthesis gas at a low level into the slurry at conditions suitable for conversion of the synthesis gas into liquid hydrocarbons, the solid catalyst particles comprising a catalytic active metal selected from cobalt or iron on a porous refractory oxide carrier, preferably selected from silica, alumina, titania, zirconia or mixtures thereof, the catalyst being present in an amount between 10 and 40 vol. percent based on total slurry volume liquids and solids, and separating liquid material from the solid catalyst particles by using a filtration system comprising an asymmetric filtration medium (the selective side at the slurry side), in which filtration system the average pressure differential over the filtration medium is at least 0.1 bar, in which process the particle size distribution is such that at least a certain amount of the catalyst particles is smaller than the average pore size of the selective layer of the filtration medium. The invention also comprises an apparatus to carry out the process described above.

  2. Merging Expertise: Preparing Collaborative Educators

    ERIC Educational Resources Information Center

    Altieri, Elizabeth M.; Colley, Kenna M.; Daniel, Leslie S.; Dickenson, Kathie W.

    2015-01-01

    In this article, teacher education faculty of a medium-size university, historically grounded in teacher preparation within a rural context, describe their ongoing work to transform their practice in order to prepare special and general educators who co-plan, co-teach, and co-assess with their counterparts in the field. Follow-up research with…

  3. Isolating "Unknown" Bacteria in the Introductory Microbiology Laboratory: A New Selective Medium for Gram-Positives.

    ERIC Educational Resources Information Center

    McKillip, John L.; Drake, MaryAnne

    1999-01-01

    Describes the development, preparation, and use of a medium that can select against a wide variety of Gram-negative bacteria while still allowing growth and differentiation of a wide range of Gram-positives. (WRM)

  4. Relocatable dense medium coal preparation plants for Indonesia

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

    Lamb, R.

    1994-12-31

    The major recent coal mine developments in Indonesia have been situated along the east coast of Kalimantan (Borneo). Design and construction in these remote areas require a high level of innovation and ingenuity to ensure that the plants can be brought on line effectively. This paper describes the design, installation, and operation of two relocatable modular dense medium plants. The plants were specifically built to overcome the difficulties of remote areas and can be assembled by a semi-skilled workforce. The two relocatable dense medium coal preparation plants recently built for mines in Kalimantan are unique in that the plants weremore » fabricated, preassembled, and wet-commissioned in Brisbane, Australia, before shipment to Indonesia. The plants are a 3OO t/h dense medium bath, cyclone, and spirals plant and a 250 t/h dense medium cyclone and spirals with reject and tailings co-disposal. The relocatable plant concept has enabled a low capital cost per ton per hour and an extremely fast construction timetable-20 weeks from contract award to completion of wet-commissioning for shipment to Indonesia.« less

  5. Cultivation of Mycobacterium bovis BCG in bioreactors.

    PubMed

    Dietrich, Guido; Mollenkopf, Hans-Joachim; Weber, Heinz; Knapp, Bernhard; Diehl, Klaus-Dieter; Hess, Jürgen; Blackkolb, Friedrich; Bröker, Michael; Kaufmann, Stefan H E; Hundt, Erika

    2002-07-03

    The Mycobacterium bovis BCG vaccine for commercial use is classically produced as surface pellicles by culture on synthetic medium. Under these conditions, reproducibility of the cultures and quality assessment are hampered by slow growth of the bacilli, the formation of bacterial aggregates and a high proportion of dead bacilli after processing and final formulation of the vaccine. Here, we established dispersed cultures of M. bovis BCG in synthetic media in small-scale bioreactors. These cultures allow recording and adjusting of culture parameters and give rise to single bacilli with a high degree of live bacteria. In the murine model, bioreactor-grown M. bovis BCG exhibited slightly stronger replication and persistence than the vaccine produced under the classical conditions. The protective efficacy against challenge with M. tuberculosis was identical for both vaccine preparations.

  6. Multifunctional substrate of Al alloy based on general hierarchical micro/nanostructures: superamphiphobicity and enhanced corrosion resistance.

    PubMed

    Li, Xuewu; Shi, Tian; Liu, Cong; Zhang, Qiaoxin; Huang, Xingjiu

    2016-10-24

    Aluminum alloys are vulnerable to penetrating and peeling failures in seawater and preparing a barrier coating to isolate the substrate from corrosive medium is an effective anticorrosion method. Inspired by the lotus leaves effect, a wetting alloy surface with enhanced anticorrosion behavior has been prepared via etch, deposition, and low-surface-energy modification. Results indicate that excellent superamphiphobicity has been achieved after the modification of the constructed hierarchical labyrinth-like microstructures and dendritic nanostructures. The as-prepared surface is also found with good chemical stability and mechanical durability. Furthermore, superior anticorrosion behaviors of the resultant samples in seawater are investigated by electrochemical measurements. Due to trapped air in micro/nanostructures, the newly presented solid-air-liquid contacting interface can help to resist the seawater penetration by greatly reducing the interface interaction between corrosive ions and the superamphiphobic surface. Finally, an optimized two-layer perceptron artificial neural network is set up to model and predict the cause-and-effect relationship between preparation conditions and the anticorrosion parameters. This work provides a great potential to extend the applications of aluminum alloys especially in marine engineering fields.

  7. Multifunctional substrate of Al alloy based on general hierarchical micro/nanostructures: superamphiphobicity and enhanced corrosion resistance

    PubMed Central

    Li, Xuewu; Shi, Tian; Liu, Cong; Zhang, Qiaoxin; Huang, Xingjiu

    2016-01-01

    Aluminum alloys are vulnerable to penetrating and peeling failures in seawater and preparing a barrier coating to isolate the substrate from corrosive medium is an effective anticorrosion method. Inspired by the lotus leaves effect, a wetting alloy surface with enhanced anticorrosion behavior has been prepared via etch, deposition, and low-surface-energy modification. Results indicate that excellent superamphiphobicity has been achieved after the modification of the constructed hierarchical labyrinth-like microstructures and dendritic nanostructures. The as-prepared surface is also found with good chemical stability and mechanical durability. Furthermore, superior anticorrosion behaviors of the resultant samples in seawater are investigated by electrochemical measurements. Due to trapped air in micro/nanostructures, the newly presented solid-air-liquid contacting interface can help to resist the seawater penetration by greatly reducing the interface interaction between corrosive ions and the superamphiphobic surface. Finally, an optimized two-layer perceptron artificial neural network is set up to model and predict the cause-and-effect relationship between preparation conditions and the anticorrosion parameters. This work provides a great potential to extend the applications of aluminum alloys especially in marine engineering fields. PMID:27775053

  8. Multifunctional substrate of Al alloy based on general hierarchical micro/nanostructures: superamphiphobicity and enhanced corrosion resistance

    NASA Astrophysics Data System (ADS)

    Li, Xuewu; Shi, Tian; Liu, Cong; Zhang, Qiaoxin; Huang, Xingjiu

    2016-10-01

    Aluminum alloys are vulnerable to penetrating and peeling failures in seawater and preparing a barrier coating to isolate the substrate from corrosive medium is an effective anticorrosion method. Inspired by the lotus leaves effect, a wetting alloy surface with enhanced anticorrosion behavior has been prepared via etch, deposition, and low-surface-energy modification. Results indicate that excellent superamphiphobicity has been achieved after the modification of the constructed hierarchical labyrinth-like microstructures and dendritic nanostructures. The as-prepared surface is also found with good chemical stability and mechanical durability. Furthermore, superior anticorrosion behaviors of the resultant samples in seawater are investigated by electrochemical measurements. Due to trapped air in micro/nanostructures, the newly presented solid-air-liquid contacting interface can help to resist the seawater penetration by greatly reducing the interface interaction between corrosive ions and the superamphiphobic surface. Finally, an optimized two-layer perceptron artificial neural network is set up to model and predict the cause-and-effect relationship between preparation conditions and the anticorrosion parameters. This work provides a great potential to extend the applications of aluminum alloys especially in marine engineering fields.

  9. pH dependent green synthesis of gold nanoparticles by completely C6-carboxylated curdlan under high temperature and various pH conditions.

    PubMed

    Qiu, Wen-Yi; Wang, Kai; Wang, Yao-Yao; Ding, Zhi-Chao; Wu, Li-Xia; Cai, Wu-Dan; Yan, Jing-Kun

    2018-01-01

    A C6-carboxylated curdlan (C6-Cc) obtained from 4-acetamido-TEMPO-mediated oxidation of curdlan was used both as a reducing and stabilizing agent for green synthesis of pH-responsive AuNPs, which was carried out by controlling the pH of the C6-Cc solution at a high temperature (100°C). C6-Cc presented a semi-flexible random coil chain in the aqueous medium at pH 5.5 and became more expanded and rigid in alkaline conditions (pH 7.1-12.0), though the primary chemical structure of C6-Cc was virtually unchanged with the pH variation. The AuNPs prepared with C6-Cc at various pHs were characterized by various instrumental measurements. The shapes and sizes of AuNPs were found to be strongly dependent on the pH of the C6-Cc solution. The C6-Cc-decorated AuNPs exhibited a more well-dispersed spherical morphology with smaller particle sizes under alkaline conditions (pH 7.1-12.0). Through this study, a facile, simple, and green method has been demonstrated for preparation of stimuli-sensitive AuNPs using biocompatible polyanionic polysaccharides. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Synthesis procedure optimization and characterization of europium (III) tungstate nanoparticles

    NASA Astrophysics Data System (ADS)

    Rahimi-Nasrabadi, Mehdi; Pourmortazavi, Seied Mahdi; Ganjali, Mohammad Reza; Reza Banan, Ali; Ahmadi, Farhad

    2014-09-01

    Taguchi robust design as a statistical method was applied for the optimization of process parameters in order to tunable, facile and fast synthesis of europium (III) tungstate nanoparticles. Europium (III) tungstate nanoparticles were synthesized by a chemical precipitation reaction involving direct addition of europium ion aqueous solution to the tungstate reagent solved in an aqueous medium. Effects of some synthesis procedure variables on the particle size of europium (III) tungstate nanoparticles were studied. Analysis of variance showed the importance of controlling tungstate concentration, cation feeding flow rate and temperature during preparation of europium (III) tungstate nanoparticles by the proposed chemical precipitation reaction. Finally, europium (III) tungstate nanoparticles were synthesized at the optimum conditions of the proposed method. The morphology and chemical composition of the prepared nano-material were characterized by means of X-ray diffraction, scanning electron microscopy, transmission electron microscopy, FT-IR spectroscopy and fluorescence.

  11. Solution-mediated cladding doping of commercial polymer optical fibers

    NASA Astrophysics Data System (ADS)

    Stajanca, Pavol; Topolniak, Ievgeniia; Pötschke, Samuel; Krebber, Katerina

    2018-03-01

    Solution doping of commercial polymethyl methacrylate (PMMA) polymer optical fibers (POFs) is presented as a novel approach for preparation of custom cladding-doped POFs (CD-POFs). The presented method is based on a solution-mediated diffusion of dopant molecules into the fiber cladding upon soaking of POFs in a methanol-dopant solution. The method was tested on three different commercial POFs using Rhodamine B as a fluorescent dopant. The dynamics of the diffusion process was studied in order to optimize the doping procedure in terms of selection of the most suitable POF, doping time and conditions. Using the optimized procedure, longer segment of fluorescent CD-POF was prepared and its performance was characterized. Fiber's potential for sensing and illumination applications was demonstrated and discussed. The proposed method represents a simple and cheap way for fabrication of custom, short to medium length CD-POFs with various dopants.

  12. BioComponent Robots

    DTIC Science & Technology

    2014-02-13

    sterile water. Culture medium was prepared with 70% Leibovitz’s L15 medium, 18% Grace’s Insect Medium, 12% fetal bovine serum (FBS), 3.4 mg/mL yeast...SECURITY CLASSIFICATION OF: The project goal was to exploit insect cell culture and tissue engineering approaches to generate biological actuators...utilizing the unique hardiness and longevity of insect cell sources for device applications for robotics. In contrast to mammalian cells and tissues

  13. Restart capability of resting-states of Euglena gracilis after 9 months of dormancy: preparation for autonomous space flight experiments

    NASA Astrophysics Data System (ADS)

    Strauch, Sebastian M.; Becker, Ina; Pölloth, Laura; Richter, Peter R.; Haag, Ferdinand W. M.; Hauslage, Jens; Lebert, Michael

    2018-04-01

    Dormant states of organisms are easier to store than the living state because they tolerate larger variations in temperature, light, storage space etc., making them attractive for laboratory culture stocks and also for experiments under special circumstances, especially space flight experiments. Like several other organisms, Euglena gracilis is capable of forming desiccation tolerant resting states in order to survive periods of unfavourable environmental conditions. In earlier experiments it was found that dormant Euglena cells must not become completely desiccated. Some residual moisture is required to ensure recovery of the resting states. To analyse the water demand in recovery of Euglena resting states, cells were transferred to a defined amount of cotton wool (0.5 g). Subsequently different volumes of medium (1, 2, 3, 4, 5, 8, 10 and 20 ml) were added in order to supply humidity; a control was set up without additional liquid. Samples were sealed in transparent 50 ml falcon tubes and stored for 9 months under three different conditions: • Constant low light conditions in a culture chamber at 20°C, • In a black box, illuminated with short light emitting diode-light pulses provided by joule thieves and • In darkness in a black box. After 9 months, cells were transferred to fresh medium and cell number, photosynthetic efficiency and movement behavior was monitored over 3 weeks. It was found that cells recovered under all conditions except in the control, where no medium was supplied. Transcription levels of 21 genes were analysed with a Multiplex-polymerase chain reaction. One hour after rehydration five of these genes were found to be up-regulated: ubiquitin, heat shock proteins HSP70, HSP90, the calcium-sensor protein frequenin and a distinct protein kinase, which is involved in gravitaxis. The results indicate a transient general stress response of the cells.

  14. Active spice-derived components can inhibit inflammatory responses of adipose tissue in obesity by suppressing inflammatory actions of macrophages and release of monocyte chemoattractant protein-1 from adipocytes.

    PubMed

    Woo, Hae-Mi; Kang, Ji-Hye; Kawada, Teruo; Yoo, Hoon; Sung, Mi-Kyung; Yu, Rina

    2007-02-13

    Inflammation plays a key role in obesity-related pathologies such as cardiovascular disease, type II diabetes, and several types of cancer. Obesity-induced inflammation entails the enhancement of the recruitment of macrophages into adipose tissue and the release of various proinflammatory proteins from fat tissue. Therefore, the modulation of inflammatory responses in obesity may be useful for preventing or ameliorating obesity-related pathologies. Some spice-derived components, which are naturally occurring phytochemicals, elicit antiobesity and antiinflammatory properties. In this study, we investigated whether active spice-derived components can be applied to the suppression of obesity-induced inflammatory responses. Mesenteric adipose tissue was isolated from obese mice fed a high-fat diet and cultured to prepare an adipose tissue-conditioned medium. Raw 264.7 macrophages were treated with the adipose tissue-conditioned medium with or without active spice-derived components (i.e., diallyl disulfide, allyl isothiocyanate, piperine, zingerone and curcumin). Chemotaxis assay was performed to measure the degree of macrophage migration. Macrophage activation was estimated by measuring tumor necrosis factor-alpha (TNF-alpha), nitric oxide, and monocyte chemoattractant protein-1 (MCP-1) concentrations. The active spice-derived components markedly suppressed the migration of macrophages induced by the mesenteric adipose tissue-conditioned medium in a dose-dependent manner. Among the active spice-derived components studied, allyl isothiocyanate, zingerone, and curcumin significantly inhibited the cellular production of proinflammatory mediators such as TNF-alpha and nitric oxide, and significantly inhibited the release of MCP-1 from 3T3-L1 adipocytes. Our findings suggest that the spice-derived components can suppress obesity-induced inflammatory responses by suppressing adipose tissue macrophage accumulation or activation and inhibiting MCP-1 release from adipocytes. These spice-derived components may have a potential to improve chronic inflammatory conditions in obesity.

  15. Conditioned medium from LS 174T goblet cells treated with oxyresveratrol strengthens tight junctions in Caco-2 cells.

    PubMed

    Hwang, Dahyun; Jo, HyunA; Hwang, Seonwook; Kim, Jeong-Keun; Kim, In-Ho; Lim, Young-Hee

    2017-01-01

    Strengthening of intestinal tight junctions provides an effective barrier from the external environment. Goblet cell-derived trefoil factor 3 (TFF3) increases transepithelial resistance by upregulating the expression of tight junction proteins. Oxyresveratrol (OXY) is a hydroxyl-substituted stilbene found in the roots, leaves, stems, and fruit of many plants and known to have various biological activities. In this study, we investigated the strengthening effect of OXY on intestinal tight junctions through stimulation of TFF production in goblet cells. We prepared conditioned medium from LS 174T goblet cells treated with OXY (GCO-CM) and investigated the effect of GCO-CM on strengthening tight junctions of Caco-2 cells. The mRNA and protein expression levels of major tight junction components (claudin-1, occludin, and ZO-1) were measured by quantitative real-time PCR and western blotting, respectively. Transepithelial electric resistance (TEER) was measured using an ohm/V meter. Monolayer permeability was evaluated by paracellular transport of fluorescein isothiocyanate-dextran. OXY showed a strong antioxidant activity. It significantly increased the expression level of TFF3 in LS 174T goblet cells. GCO-CM prepared by treatment with 2.5, 5, and 10μg/ml OXY did not show cytotoxicity in Caco-2 cells. GCO-CM increased the mRNA and protein expression levels of claudin-1, occludin, and ZO-1. It also significantly increased tight junction integrity and reduced permeability in a dose-dependent manner. OXY stimulates the expression of TFF3 in goblet cells, which might increase the integrity of the intestinal tight junction barrier. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  16. Controlling silver nanoparticle exposure in algal toxicity testing – A matter of timing

    PubMed Central

    Baun, Anders

    2015-01-01

    The aquatic ecotoxicity testing of nanoparticles is complicated by unstable exposure conditions resulting from various transformation processes of nanoparticles in aqueous suspensions. In this study, we investigated the influence of exposure timing on the algal test response to silver nanoparticles (AgNPs), by reducing the incubation time and by aging the AgNPs in algal medium prior to testing. The freshwater green algae Pseudokirchneriella subcapitata were exposed to AgNO3, NM-300 K (a representative AgNP) and citrate stabilized AgNPs from two different manufacturers (AgNP1 and AgNP2) in a standard algal growth inhibition test (ISO 8692:2004) for 48 h and a short-term (2 h) 14C-assimilation test. For AgNO3, similar responses were obtained in the two tests, whereas freshly prepared suspensions of citrate stabilized AgNPs were less toxic in the 2-h tests compared to the 48-h tests. The 2-h test was found applicable for dissolved silver, but yielded non-monotonous concentration–response relationships and poor reproducibility for freshly prepared AgNP suspensions. However, when aging AgNPs in algal medium 24 h prior to testing, clear concentration–response patterns emerged and reproducibility increased. Prolonged aging to 48 h increased toxicity in the 2-h tests whereas aging beyond 48 h reduced toxicity. Our results demonstrate that the outcome of algal toxicity testing of AgNPs is highly influenced not only by the test duration, but also by the time passed from the moment AgNPs are added to the test medium. This time-dependency should be considered when nanomaterial dispersion protocols for ecotoxicity testing are developed. PMID:24842597

  17. 40 CFR 205.57-3 - Test vehicle preparation.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Test vehicle preparation. 205.57-3... PROGRAMS TRANSPORTATION EQUIPMENT NOISE EMISSION CONTROLS Medium and Heavy Trucks § 205.57-3 Test vehicle preparation. (a) Prior to the official test, the test vehicle selected in accordance with § 205-57-2 shall not...

  18. 47 CFR 42.5 - Preparation and preservation of reproductions of original records.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 3 2010-10-01 2010-10-01 false Preparation and preservation of reproductions... Instructions § 42.5 Preparation and preservation of reproductions of original records. (a) Each carrier may use... medium shall be accompanied by a statement clearly indicating the type of data included in the record and...

  19. 47 CFR 42.5 - Preparation and preservation of reproductions of original records.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 47 Telecommunication 3 2011-10-01 2011-10-01 false Preparation and preservation of reproductions... Instructions § 42.5 Preparation and preservation of reproductions of original records. (a) Each carrier may use... medium shall be accompanied by a statement clearly indicating the type of data included in the record and...

  20. [Nutritional value of daily diets prepared in several regions of the country. Part V. Value of group b vitamins].

    PubMed

    Nadolna, I; Wiśniewska, W; Kunachowicz, H

    1991-01-01

    Studies of the content of group B vitamins: thiamin, riboflavin, niacin and vitamin b6 in daily diets of manual and mental workers with medium income were carried on. The diets were prepared for five regions of the country (Warszawa, Lublin, Olsztyn, Poznań, Wrocław) under laboratory conditions. These diets contained respectively: thiamin 1.17 and 1.11 mg, riboflavin 1.48 and 1.55 mg; niacin 11.3 and 10.2 mg and vitamin B6 1.50 and 1.21 mg per day. According to the studies the realization of daily requirements by these diets for thiamin were met 82 and 98%, for niacin 62 and 65%, and for vitamin B6 79 and 64%. The comparison of the presently studied diets with the ones from 1973, 1980 and 1981 showed that the content of niacin and vitamin B6 intakes systematically decrease. There were no considerable differences in the contents of group B vitamins between diets prepared in all five regions of Poland.

  1. Enantioselective Synthesis of Various Cyanohydrins Using Covalently Immobilized Preparations of Hydroxynitrile Lyase from Prunus dulcis.

    PubMed

    Alagöz, Dilek; Tükel, S Seyhan; Yildirim, Deniz

    2015-11-01

    The carrier-based and carrier-free (cross-linked enzyme aggregate) covalent immobilizations of Prunus dulcis hydroxynitrile lyase were investigated. The immobilized preparations were tested for enantioselective carbon-carbon bond formation activity in the biphasic medium. Of the tested preparations, only cross-linked enzyme aggregate of P. dulcis hydroxynitrile lyase (PdHNL-CLEA) achieved the synthesis of (R)-mandelonitrile with 93% yield and 99% enantiopurity. PdHNL-CLEA was also used in the synthesis of various (R)-cyanohydrins from corresponding aldehydes/ketones and hydrocyanic acid. When 4-methoxybenzaldehyde, 4-methyl benzaldehyde, and 4-hydroxybenzaldehyde were used as substrates, the yield-enantiomeric excess of corresponding (R)-cyanohydrins were obtained as 95-95, 85-79, and 2-25%, respectively, after 96 h at pH 4.0 and 5 °C. For acetophenone, 4-fluoroacetophenone, 4-chloroacetophenone, 4-bromoacetophenone, and 4-iodoacetophenone, the yield-enantiomeric excess of corresponding (R)-cyanohydrins were 1-99, 20-84, 11-95, 5-99, and 3-24%, respectively at the same conditions. The results demonstrate PdHNL-CLEA can be effectively used in the synthesis of (R)-mandelonitrile.

  2. Improved Chemically Defined Basal Medium (CMRL-1969) for Primary Monkey Kidney and Human Diploid Cells 1

    PubMed Central

    Healy, G. M.; Teleki, S.; Seefried, A. V.; Walton, M. J.; Macmorine, H. G.

    1971-01-01

    An improved tissue culture basal medium, CMRL-1969, supplemented with serum, has been evaluated by measuring the growth responses of primary cultures of trypsin-dispersed monkey kidney cells (PMKC) and of an established culture of a human diploid cell strain (HDCS). Medium H597, an early modification of medium 199 which has been used successfully in the preparation of poliomyelitis vaccine for 15 years, was used for comparison. In addition, parallel testing was done with Basal Medium Eagle (BME) widely used for the growth of HDCS. The improvements in basal medium CMRL-1969 are attributed to changes in amino acid concentrations, in vitamin composition, and, in particular, to enhanced buffering capacity. The latter has been achieved by the use of free-base amino acids and by increasing the dibasic sodium phosphate. The new medium has already been used to advantage for the production of polioviruses in PMKC where equivalent titers were obtained from cultures initiated with 70% of the number of cells required with earlier media. The population-doubling time was reduced in this system. Also, with small inocula of HDCS, the time required to obtain maximum cell yield was shorter with CMRL-1969 than with BME. Both media were supplemented with 10% calf serum. Maximum cell yields after repeated subcultivation in the new basal medium were greatly increased and the stability of the strain, as shown by chromosomal analysis, was not affected. Basal medium CMRL-1969 can be prepared easily in liquid or powdered form. PMID:4322279

  3. Enzymatic preparation of structured oils containing short-chain fatty acids.

    PubMed

    Kanda, Ayato; Namiki, Fusako; Hara, Setsuko

    2010-01-01

    Structured oils prepared by enzymatic transacylation with triacylglycerols (TAGs) and various fatty acids (FAs) were characterized. Transacylation with trilaurin and saturated FAs (C4:0-C16:0) was performed using Lipozyme RM-IM under standard reaction conditions. The structured oils thus produced had transacylation ratios of 25-37%, as medium-chain FAs > long-chain FAs > short-chain FAs. This result confirmed that short-chain FAs have little reactivity in enzymatic transacylation. All prepared oils shared the same composition of TAG molecular species, as demonstrated by HPLC analysis, and contained a mixture of mono-substituted, di-substituted, and non-substituted TAGs. The reaction conditions for transacylation with TAGs and short-chain FAs were optimized to improve transacylation ratios. The introduction ratios of C4:0, C5:0, and C6:0 into trilaurin were increased to 52.4, 42.5, and 34.1%, respectively, by extending the reaction time. Transacylation between TAGs and short-chain FAs was further examined by using Lipase PL. C4:0 was introduced at 51.1%, the same ratio as for Lipozyme RM-IM. When C5:0 and C6:0 were used as the FA substrate, the transacylation ratios obtained were 47.7 and 43.4%, respectively, higher than those for Lipase RM-IM. Lipase PL is therefore useful for introducing short-chain FAs into TAGs.

  4. Bioremediation of coal contaminated soil under sulfate-reducing condition.

    PubMed

    Kuwano, Y; Shimizu, Y

    2006-01-01

    The objective of this study was to investigate the biodegradation of coal-derived hydrocarbons, especially high molecular weight (HMW) components, under anaerobic conditions. For this purpose biodegradation experiments were performed, using specifically designed soil column bioreactors. For the experiment, coal-contaminated soil was prepared, which contains high molecular weight hydrocarbons at high concentration (approx. 55.5 mgC g-drysoil(-1)). The experiment was carried out in two different conditions: sulfate reducing (SR) condition (SO4(2-) = 10 mmol l(-1) in the liquid medium) and control condition (SO4(2-)<0.5 mmol l(-1)). Although no degradation was observed under the control condition, the resin fraction decreased to half (from 6,541 to 3,386 mgC g-soil(-1)) under SR condition, with the concomitant increase of two PAHs (phenanthrene and fluoranthene, 9 and 2.5 times, respectively). From these results, we could conclude that high molecular hydrocarbons were biodegradable and transformed to low molecular weight PAHs under the sulfate-reducing condition. Since these PAHs are known to be biologically degraded under aerobic condition, a serial combination of anaerobic (sulfate reducing) and then aerobic bioremediations could be effective and useful for the soil pollution by petroleum and/or coal derived hydrocarbons.

  5. Monitoring sulfide and sulfate-reducing bacteria

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

    Tanner, R.S.

    1995-12-31

    Simple yet precise and accurate methods for monitoring sulfate-reducing bacteria (SRB) and sulfide remain useful for the study of bacterial souring and corrosion. Test kits are available to measure sulfide in field samples. A more precise methylene blue sulfide assay for both field and laboratory studies is described here. Improved media, compared to that in API RP-38, for enumeration of SRB have been formulated. One of these, API-RST, contained cysteine (1.1 mM) as a reducing agent, which may be a confounding source of sulfide. While cysteine was required for rapid enumeration of SRB from environmental samples, the concentration of cysteinemore » in medium could be reduced to 0.4 mM. It was also determined that elevated levels of yeast extract (>1 g/liter) could interfere with enumeration of SRB from environmental samples. The API-RST medium was modified to a RST-11 medium. Other changes in medium composition, in addition to reduction of cysteine, included reduction of the concentration of phosphate from 3.4 mM to 2.2 mM, reduction of the concentration of ferrous iron from 0.8 mM to 0.5 mM and preparation of a stock mineral solution to ease medium preparation. SRB from environmental samples could be enumerated in a week in this medium.« less

  6. On pleasure and thrill: the interplay between arousal and valence during visual word recognition.

    PubMed

    Recio, Guillermo; Conrad, Markus; Hansen, Laura B; Jacobs, Arthur M

    2014-07-01

    We investigated the interplay between arousal and valence in the early processing of affective words. Event-related potentials (ERPs) were recorded while participants read words organized in an orthogonal design with the factors valence (positive, negative, neutral) and arousal (low, medium, high) in a lexical decision task. We observed faster reaction times for words of positive valence and for those of high arousal. Data from ERPs showed increased early posterior negativity (EPN) suggesting improved visual processing of these conditions. Valence effects appeared for medium and low arousal and were absent for high arousal. Arousal effects were obtained for neutral and negative words but were absent for positive words. These results suggest independent contributions of arousal and valence at early attentional stages of processing. Arousal effects preceded valence effects in the ERP data suggesting that arousal serves as an early alert system preparing a subsequent evaluation in terms of valence. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. Synthesis of Azanucleosides by Anodic Oxidation in a Lithium Perchlorate-Nitroalkane Medium and Diversification at the 4'-Nitrogen Position.

    PubMed

    Shoji, Takao; Kim, Shokaku; Chiba, Kazuhiro

    2017-03-27

    Azanucleosides, in which the 4'-oxygen atom has been replaced with a nitrogen atom, have drawn much attention owing to their anticancer and antivirus activity, and tolerance towards nucleases. However, the traditional synthetic strategy requires multiple steps and harsh conditions, thereby limiting the structural and functional diversity of the products. Herein we describe the synthesis of azanucleosides by an electrochemical reaction in a lithium perchlorate-nitroethane medium, followed by postmodification at the 4'-N position. N-Acryloyl prolinol derivatives were converted into azanucleosides by anodic activation of the N-α-C-H bond. Moreover, the use of nitroethane instead of nitromethane lowered the oxidation potential of the N-acryloyl prolinols and increased the Faradic yield. The prepared azanucleosides were efficiently functionalized at the 4'-N-acryloyl group with a lipophilic alkanethiol and a fluorescent dye by conjugate addition and olefin cross-metathesis, respectively. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Electron and Fluorescence Microscopy of Extracellular Glucan and Aryl-Alcohol Oxidase during Wheat-Straw Degradation by Pleurotus eryngii

    PubMed Central

    Barrasa, J. M.; Gutiérrez, A.; Escaso, V.; Guillén, F.; Martínez, M. J.; Martínez, A. T.

    1998-01-01

    The ligninolytic fungus Pleurotus eryngii grown in liquid medium secreted extracellular polysaccharide (87% glucose) and the H2O2-producing enzyme aryl-alcohol oxidase (AAO). The production of both was stimulated by wheat-straw. Polyclonal antibodies against purified AAO were obtained, and a complex of glucanase and colloidal gold was prepared. With these tools, the localization of AAO and extracellular glucan in mycelium from liquid medium and straw degraded under solid-state fermentation conditions was investigated by transmission electron microscopy (TEM) and fluorescence microscopy. These studies revealed that P. eryngii produces a hyphal sheath consisting of a thin glucan layer. This sheath appeared to be involved in both mycelial adhesion to the straw cell wall during degradation and AAO immobilization on hyphal surfaces, with the latter evidenced by double labeling. AAO distribution during differential degradation of straw tissues was observed by immunofluorescence microscopy. Finally, TEM immunogold studies confirmed that AAO penetrates the plant cell wall during P. eryngii degradation of wheat straw. PMID:9435085

  9. Recombinant von Willebrand factor: preclinical development.

    PubMed

    Plaimauer, B; Schlokat, U; Turecek, P L; Mitterer, A; Mundt, W; Auer, W; Pichler, L; Gritsch, H; Schwarz, H P

    2001-08-01

    Von Willebrand factor (vWF) is a multimeric glycoprotein (GP) that attracts platelets to the site of vascular injury, mediates platelet-platelet interaction, and stabilizes factor VIII (FVIII) in the circulation. Quantitative and qualitative defects of vWF result in von Willebrand disease (vWD), manifested by modest to severe bleeding episodes. Substitution therapy, with plasma-derived FVIII/vWF complex concentrates, is used for patients suffering the more severe forms of vWD. Efficacy of these preparations is often unsatisfactory because inadvertent proteolytic degradation during the manufacturing process causes them to lack the hemostatically most active high-molecular-weight multimers. In contrast, recombinant vWF (r-vWF), which is constitutively expressed at high yields in Chinese hamster ovary (CHO) cells and secreted into the conditioned medium under perfusion fermentation in "protein-free" medium, has high-molecular-weight multimers of extraordinary structural integrity. Functional analysis has shown that r-vWF promotes ristocetin cofactor-mediated platelet aggregation, collagen interaction and FVIII binding, and platelet-collagen adhesion under shear stress. Infusing vWF-deficient animals with r-vWF corrected vWF concentration and reduced blood loss, subsequently stabilizing endogenous FVIII associated with the reduction of bleeding time. Compared with plasma-derived vWF preparations, r-vWF was found to have a prolonged half-life, further enhancing the potential value of r-vWF as a therapeutic agent for treating patients suffering from vWD.

  10. Two-dimensional preparative liquid chromatography system for preparative separation of minor amount components from complicated natural products.

    PubMed

    Qiu, Ying-Kun; Chen, Fang-Fang; Zhang, Ling-Ling; Yan, Xia; Chen, Lin; Fang, Mei-Juan; Wu, Zhen

    2014-04-11

    An on-line comprehensive two-dimensional preparative liquid chromatography system was developed for preparative separation of minor amount components from complicated natural products. Medium-pressure liquid chromatograph (MPLC) was applied as the first dimension and preparative HPLC as the second one, in conjunction with trapping column and makeup pump. The performance of the trapping column was evaluated, in terms of column size, dilution ratio and diameter-height ratio, as well as system pressure from the view of medium pressure liquid chromatograph. Satisfactory trapping efficiency can be achieved using a commercially available 15 mm × 30 mm i.d. ODS pre-column. The instrument operation and the performance of this MPLC×preparative HPLC system were illustrated by gram-scale isolation of crude macro-porous resin enriched water extract of Rheum hotaoense. Automated multi-step preparative separation of 25 compounds, whose structures were identified by MS, (1)H NMR and even by less-sensitive (13)C NMR, could be achieved in a short period of time using this system, exhibiting great advantages in analytical efficiency and sample treatment capacity compared with conventional methods. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Formulation and evaluation of a montelukast sodium orally disintegrating tablet with a similar dissolution profile as the marketed product.

    PubMed

    Chen, Yong; Feng, Tingting; Li, Yong; Du, Bin; Weng, Weiyu

    2017-03-01

    A major challenge of orally disintegrating tablet (ODT) development is predicting its bioequivalence to its corresponding marketed product. Therefore, comparing ODT dissolution profiles to those of the corresponding marketed product is very important. The objective of this study was to develop a 5.2-mg montelukast sodium (MS) ODT with a similar dissolution profile to that of the marketed chewable tablet. Dissolution profiles were examined in different media to screen each formulation. We found that MS dissolution from ODTs in acidic medium heavily depended on manufacturing methods. All MS ODTs prepared using direct compression rapidly disintegrated in acidic medium. However, dispersed MS powders aggregated into sticky masses, resulting in slow dissolution. In contrast, MS ODTs prepared using wet granulation had much faster dissolution rates in acidic medium with no obvious aggregation. Additionally, the optimized formulation, prepared using wet granulation, displayed similar dissolution profiles to the marketed reference in all four types of media examined (f 2  >   50). The in vitro disintegration time of the optimized ODT was 9.5 ± 2.4 s, which meets FDA requirements. In conclusion, the wet granulation preparation method of MS ODTs resulted in a product with equivalent dissolution profiles as those of the marketed product.

  12. Whole cell entrapment techniques.

    PubMed

    Trelles, Jorge A; Rivero, Cintia W

    2013-01-01

    Microbial whole cells are efficient, ecological, and low-cost catalysts that have been successfully applied in the pharmaceutical, environmental, and alimentary industries, among others. Microorganism immobilization is a good way to carry out the bioprocess under preparative conditions. The main advantages of this methodology lie in their high operational stability, easy upstream separation and bioprocess scale-up feasibility. Cell entrapment is the most widely used technique for whole cell immobilization. This technique-in which the cells are included within a rigid network-is porous enough to allow the diffusion of substrates and products, protects the selected microorganism from the reaction medium, and has high immobilization efficiency (100 % in most cases).

  13. An ultrasensitive and selective electrochemical sensor for determination of estrone 3-sulfate sodium salt based on molecularly imprinted polymer modified carbon paste electrode.

    PubMed

    Song, Han; Wang, Yuli; Zhang, Lu; Tian, Liping; Luo, Jun; Zhao, Na; Han, Yajie; Zhao, Feilang; Ying, Xue; Li, Yingchun

    2017-11-01

    A highly sensitive and selective electrochemical sensor based on carbon paste electrode (CPE) modified with molecularly imprinted polymers (MIPs) has been developed for the determination of estrone 3-sulfate sodium salt (ESS). MIPs were prepared in polar medium via bulk polymerization and characterized by scanning electron microscopy and infrared spectroscopy. Cyclic voltammetry was performed to the study preparation process and binding behavior of the MIP-modified CPE (MIP/CPE) toward ESS. The conditions for preparing MIPs and MIP/CPE as well as ESS detection were optimized. Under the optimal experimental conditions, the detection linear range for ESS is 4 × 10 -12 to 6 × 10 -9  M with a limit of detection of 1.18 × 10 -12  M (S/N = 3). In addition, the sensor exhibits high binding affinity toward ESS over its structural analogues with excellent repeatability and stability. The fabricated MIP/CPE was then successfully employed to detect ESS in pregnant mare urine (PMU) without any pretreatment, and the average recoveries were from 99.6 to 104.9% with relative standard deviation less than 3.0%. High-performance liquid chromatography was adopted as a reference to validate the established approach in detecting ESS and their results showed good agreement. The as-prepared sensor has high potential to be a decent tool for on-site determination of ESS in PMU in a fast and convenient manner. Graphical Abstract ᅟ.

  14. Development of a Suitable Dissolution Method for the Combined Tablet Formulation of Atorvastatin and Ezetimibe by RP-LC Method.

    PubMed

    Ozkan Cansel, Kose; Ozgur, Esim; Sevinc, Kurbanoglu; Ayhan, Savaser; Ozkan, Sibel A; Yalcin, Ozkan

    2016-01-01

    Pharmaceutical preparations of ezetimibe and atorvastatin are generally used to regulate the lipid level in blood. It decreases the secondary events for patients with high cholesterol and clinical cardiovascular disease such as non-fatal or fatal heart attack. There is no any pharmacopoeia method available for the dissolution testing recommended by the FDA. Development of dissolution tests method is very critical parameter especially for the pharmaceutical preparations that contain Class II drugs (slightly soluble, good permeable). In the proposed method, the effects of pH and surfactant on the dissolution of poorly water soluble combined drug therapy with a different pKa values in an in vitro environment is investigated. The content of our study was designed to answer these open-ended questions. The optimized test conditions achieved under sink conditions with USP apparatus 2 at a paddle rotation speed of 75 rpm and 900 ml in 0.01 M Acetate buffer (pH= 6.8) containing 0.45% SDS as a dissolution medium. Quantification of dissolution samples were analyzed with a new fully validated RP-LC method with UV detection at 242 nm.

  15. Silver/poly(vinyl alcohol) nanocomposite film prepared using water in oil microemulsion for antibacterial applications.

    PubMed

    Fatema, Ummul K; Rahman, M Muhibur; Islam, M Rakibul; Mollah, M Yousuf A; Susan, Md Abu Bin Hasan

    2018-03-15

    Water in oil microemulsion (w/o) is a simple preparative route for nanoparticles where water droplets (dispersed in continuous oil medium and stabilized by surfactants and cosurfactants) act as nanoreactors to carry out chemical reactions. If polymeric matrix is incorporated inside the core of the microemulsions, it should prevent the agglomeration of nanoparticles after separation from microemulsions. Thus polymer nanocomposite films prepared from w/o microemulsions are expected to give narrow and homogeneous size distribution of nanoparticles throughout the polymer host. Silver/poly(vinyl alcohol) (Ag/PVA) nanocomposite film was successfully prepared, for the first time, using Triton X-100 (TX-100)/1-butanol/cyclohexane/water microemulsion. Reduction of the metal salt was carried out in the core of w/o microemulsion droplets containing PVA polymeric matrix. After separation from the microemulsion, Ag/PVA nanocomposite film was then prepared by solution casting method. The antibacterial activity of the nanocomposites was tested against Gram-negative, Escherichia coli and Gram-positive, Staphylococcus aureus by agar diffusion method. Ag nanoparticles with an average diameter of 105 nm could be synthesized using PVA, whereas in the absence of PVA the nanoparticles agglomerated. The distribution of Ag nanoparticles on PVA surface of the nanocomposite film prepared using microemulsion was uniform, whereas the film prepared through in situ generation of Ag nanoparticles by chemical reduction process on PVA host showed non-uniform, coagulated, bunches of Ag nanoparticles. The film synthesized using microemulsion exhibited enhanced antibacterial efficacy compared to that prepared through in situ synthesis under the same test condition. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. A new method of preparing embeddment-free sections for transmission electron microscopy: applications to the cytoskeletal framework and other three-dimensional networks.

    PubMed

    Capco, D G; Krochmalnic, G; Penman, S

    1984-05-01

    Diethylene glycol distearate is used as a removable embedding medium to produce embeddment -free sections for transmission electron microscopy. The easily cut sections of this material float and form ribbons in a water-filled knife trough and exhibit interference colors that aid in the selection of sections of equal thickness. The images obtained with embeddment -free sections are compared with those from the more conventional epoxy-embedded sections, and illustrate that embedding medium can obscure important biological structures, especially protein filament networks. The embeddment -free section methodology is well suited for morphological studies of cytoskeletal preparations obtained by extraction of cells with nonionic detergent in cytoskeletal stabilizing medium. The embeddment -free section also serves to bridge the very different images afforded by embedded sections and unembedded whole mounts.

  17. Technique for restoration of mite (Acari) preparations in deteriorated Hoyer's medium.

    PubMed

    Jacinavicius, F C; Badari, J C; Ramirez, D G; Moraes, R H P; Onofrio, V C; Barros-Battesti, D M

    2013-06-01

    The Acari Collection of Instituto Butantan (IBSP), São Paulo, Brazil, includes many types and other identified mite specimens that were mounted in Hoyer's medium, mainly in the first part of last century. An effort to restore degraded preparations was initiated in 1996. In this process, an improved technique was developed, allowing the adequate cleaning of specimens mounted up to 50-70 years before. Types and other identified specimens of Trombidiformes (Harpirhynchidae and Trombiculidae), Sarcoptiformes (Acaridae, Atopomelidae, Listrophoridae, and Psoroptidae) and Mesostigmata (Dermanyssidae, Ixodorhynchidae, Laelapidae, Macronyssidae, and Spinturnicidae) deposited at IBSP Collection have been satisfactorily restored.

  18. Kitchen Microbiology: It's Easier Than You Think!

    ERIC Educational Resources Information Center

    Wilcoxson, Catherine; Shand, Stacey M.; Shand, Richard F.

    1999-01-01

    Offers a simple experiment that tests the effectiveness of cleaners that inhibit bacterial growth. Outlines a step-by-step method for preparing culture media that can be used for isolation and propagation of microorganisms. Concludes that kitchen medium is as efficient as the professionally-made medium in supporting colony growth. (CCM)

  19. Computational method to predict thermodynamic, transport, and flow properties for the modified Langley 8-foot high-temperature tunnel

    NASA Technical Reports Server (NTRS)

    Venkateswaran, S.; Hunt, L. Roane; Prabhu, Ramadas K.

    1992-01-01

    The Langley 8 foot high temperature tunnel (8 ft HTT) is used to test components of hypersonic vehicles for aerothermal loads definition and structural component verification. The test medium of the 8 ft HTT is obtained by burning a mixture of methane and air under high pressure; the combustion products are expanded through an axisymmetric conical contoured nozzle to simulate atmospheric flight at Mach 7. This facility was modified to raise the oxygen content of the test medium to match that of air and to include Mach 4 and Mach 5 capabilities. These modifications will facilitate the testing of hypersonic air breathing propulsion systems for a wide range of flight conditions. A computational method to predict the thermodynamic, transport, and flow properties of the equilibrium chemically reacting oxygen enriched methane-air combustion products was implemented in a computer code. This code calculates the fuel, air, and oxygen mass flow rates and test section flow properties for Mach 7, 5, and 4 nozzle configurations for given combustor and mixer conditions. Salient features of the 8 ft HTT are described, and some of the predicted tunnel operational characteristics are presented in the carpet plots to assist users in preparing test plans.

  20. Vibrational Spectroscopy after OSU - From C2- to Interstellar Polycyclic Aromatic Hydrocarbons

    NASA Technical Reports Server (NTRS)

    Allamandola, Louis J.

    2006-01-01

    The composition of interstellar ice and dust provides insight into the chemical history of the interstellar medium and early solar system. It is now possible to probe this unique and unusual chemistry and determine the composition of these microscopic interstellar particles which are hundreds to many thousands of light years away thanks to substantial progress in two areas: astronomical spectroscopic techniques in the middle-infrared, the spectral region most diagnostic of chemical composition, and laboratory simulations which realistically reproduce the critical conditions in various interstellar environments. High quality infrared spectra of many different astronomical sources, some associated with giant, dark molecular clouds -the birthplace of stars and planets- and others in more tenuous, UV radiation rich regions are now available. The fundamentals of IR spectroscopy and what comparisons of astronomical IR spectra with laboratory spectra of materials prepared under realistic simulated interstellar conditions tell us about the components of these materials is the subject of this talk. These observations have shown that mixed molecular ices comprised of H2O, CH3OH, CO, NH3 and H2CO contain most of the molecular material in molecular clouds and that gas phase, ionized polycyclic aromatic hydrocarbons (PAHs) are widespread and surprisingly abundant throughout most of the interstellar medium.

  1. Protective properties of ginsenoside Rb1 against UV-B radiation-induced oxidative stress in human dermal keratinocytes.

    PubMed

    Oh, Sun-Joo; Kim, Kyunghoon; Lim, Chang-Jin

    2015-06-01

    Ginsenosides, also known as ginseng saponins, are responsible for most pharmacological effect of ginseng. Ginsenoside Rb1 (Rb1) exerts a variety of pharmacological properties, including anti-inflammatory, antistress, anti-aging and anti-neurodegenerative activities. The aim of the present work was to assess the skin anti-photoaging properties of Rb1 in human dermal keratinocyte HaCaT cells. The anti-photoaging activity was evaluated by analyzing the levels of reactive oxygen species (ROS) and matrix metalloproteinases (MMPs) as well as cell viability for HaCaT cells under UV-B irradiation. Rb1 was able to suppress the ROS levels which were elevated under UV-B irradiation, and unable to influence cellular survival in UV-B-irradiated HaCaT cells. Rb1 diminished the enhancement of MMP-2 gelatinolytic activity in conditioned medium, which corresponded with the decreased MMP-2 protein levels in both conditioned medium and cellular lysate prepared from UV-B-irradiated HaCaT cultures. Rb1 could restore the total glutathione (GSH) and superoxide dismutase (SOD) activity diminished in UV-B-irradiated HaCaT cells. Ginsenoside Rb1 possesses skin anti-photoaging properties through scavenging ROS and decreasing MMP-2 levels possibly by enhancing antioxidant activity in keratinocytes under UV-B irradiation.

  2. Hypoxic-Preconditioned Bone Marrow Stem Cell Medium Significantly Improves Outcome After Retinal Ischemia in Rats

    PubMed Central

    Roth, Steven; Dreixler, John C.; Mathew, Biji; Balyasnikova, Irina; Mann, Jacob R.; Boddapati, Venkat; Xue, Lai; Lesniak, Maciej S.

    2016-01-01

    Purpose We have previously demonstrated the protective effect of bone marrow stem cell (BMSC)-conditioned medium in retinal ischemic injury. We hypothesized here that hypoxic preconditioning of stem cells significantly enhances the neuroprotective effect of the conditioned medium and thereby augments the protective effect in ischemic retina. Methods Rats were subjected to retinal ischemia by increasing intraocular pressure to 130 to 135 mm Hg for 55 minutes. Hypoxic-preconditioned, hypoxic unconditioned, or normoxic medium was injected into the vitreous 24 hours after ischemia ended. Recovery was assessed 7 days after injections by comparing electroretinography measurements, histologic examination, and apoptosis (TUNEL, terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling assay). To compare proteins secreted into the medium in the groups and the effect of hypoxic exposure, we used rat cytokine arrays. Results Eyes injected with hypoxic BMSC–conditioned medium 24 hours after ischemia demonstrated significantly enhanced return of retinal function, decreased retinal ganglion cell layer loss, and attenuated apoptosis compared to those administered normoxic or hypoxic unconditioned medium. Hypoxic-preconditioned medium had 21 significantly increased protein levels compared to normoxic medium. Conclusions The medium from hypoxic-preconditioned BMSCs robustly restored retinal function and prevented cell loss after ischemia when injected 24 hours after ischemia. The protective effect was even more pronounced than in our previous studies of normoxic conditioned medium. Prosurvival signals triggered by the secretome may play a role in this neuroprotective effect. PMID:27367588

  3. Biomimetic evaluation of β tricalcium phosphate prepared by hot isostatic pressing

    PubMed Central

    Mateescu, Mihaela; Rguitti, Emmanuelle; Ponche, Arnaud; Descamps, Michel; Anselme, Karine

    2012-01-01

    Two types of completely densified β-TCP tablets were synthesized from a stoichiometric β-TCP powder. The first ones (TCP) were conventionally sintered, while the second ones (TCP-T) were sintered and treated by hot isostatic process (HIP). The HIP produced completely densified materials with relative densities greater than 99.9% and a transparent appearance of tablets. Samples were immersed in culture medium with (CM) or without serum (NCM) in static and dynamic conditions for a biomimetic evaluation. Similarly, SaOs-2 cells were cultured on samples in a static or dynamic flow perfusion system. The results of surface transformation in absence of cells showed that the dynamic condition increased the speed of calcium phosphate precipitations compared with the static condition. The morphology of precipitates was different with nature of tablets. The immersion in CM did impede this precipitation. XPS analysis of TCP-T tablets showed the presence of hydroxyapatite (HA) precipitates after incubation in NCM while octacalcium phosphate (OCP) precipitates were formed after incubation in CM. The analysis of the response of SaOs-2 cells on surfaces showed that the two types of materials are biocompatible. However, the dynamic mode of culture stimulated the differentiation of cells. Finally, it appears that the HIP treatment of TCP produces highly densified and transparent samples that display a good in vitro biocompatibility in static and dynamic culture conditions. Moreover, an interesting result of this work is the relationship between the presence of proteins in the immersion medium and the quality of precipitates formed on hipped TCP surface. PMID:23507861

  4. Zinc is released by cultured astrocytes as a gliotransmitter under hypoosmotic stress-loaded conditions and regulates microglial activity.

    PubMed

    Segawa, Shohei; Nishiura, Takeshi; Furuta, Takahiro; Ohsato, Yuki; Tani, Misaki; Nishida, Kentaro; Nagasawa, Kazuki

    2014-01-17

    Astrocytes contribute to the maintenance of brain homeostasis via the release of gliotransmitters such as ATP and glutamate. Here we examined whether zinc was released from astrocytes under stress-loaded conditions, and was involved in the regulation of microglial activity as a gliotransmitter. Hypoosmotic stress was loaded to astrocytes using balanced salt solution prepared to 214-314 mOsmol/L, and then intra- and extra-cellular zinc levels were assessed using Newport Green DCF diacetate (NG) and ICP-MS, respectively. Microglial activation by the astrocytic supernatant was assessed by their morphological changes and poly(ADP-ribose) (PAR) polymer accumulation. Exposure of astrocytes to hypoosmotic buffer, increased the extracellular ATP level in osmolarity-dependent manners, indicating a load of hypoosmotic stress. In hypoosmotic stress-loaded astrocytes, there were apparent increases in the intra- and extra-cellular zinc levels. Incubation of microglia in the astrocytic conditioned medium transformed them into the activated "amoeboid" form and induced PAR formation. Administration of an extracellular zinc chelator, CaEDTA, to the astrocytic conditioned medium almost completely prevented the microglial activation. Treatment of astrocytes with an intracellular zinc chelator, TPEN, suppressed the hypoosmotic stress-increased intracellular, but not the extracellular, zinc level, and the increase in the intracellular zinc level was blocked partially by a nitric oxide synthase inhibitor, but not by CaEDTA, indicating that the mechanisms underlying the increases in the intra- and extra-cellular zinc levels might be different. These findings suggest that under hypoosmotic stress-loaded conditions, zinc is released from astrocytes and then plays a primary role in microglial activation as a gliotransmitter. Copyright © 2013 Elsevier Inc. All rights reserved.

  5. Water-in-oil Pickering emulsions stabilized by stearoylated microcrystalline cellulose.

    PubMed

    Pang, Bo; Liu, Huan; Liu, Peiwen; Peng, Xinwen; Zhang, Kai

    2018-03-01

    Hydrophobic particles with static water contact angles larger than 90° are more like to stabilize W/O Pickering emulsions. In particular, high internal phase Pickering emulsions (HIPEs) are of great interest for diverse applications. However, W/O HIPEs have rarely been realized using sustainable biopolymers. Herein, we used stearoylated microcrystalline cellulose (SMCC) to stabilize W/O Pickering emulsions and especially, W/O HIPEs. Moreover, these W/O HIPEs can be further used as platforms for the preparation of porous materials, such as porous foams. Stearoylated microcrystalline cellulose (SMCC) was prepared by modifying MCC with stearoyl chloride under heterogeneous conditions. Using SMCC as emulsifiers, W/O medium and high internal phase Pickering emulsions (MIPEs and HIPEs) with various organic solvents as continuous phases were prepared using one-step and two-step methods, respectively. Polystyrene (PS) foams were prepared after polymerization of oil phase using HIPEs as templates and their oil/water separation capacity were studied. SMCC could efficiently stabilize W/O Pickering emulsions and HIPEs could only be prepared via the two-step method. The internal phase volume fraction of the SMCC-stabilized HIPEs reached as high as 89%. Diverse internal phase volume fractions led to distinct inner structures of foams with closed or open cells. These macroporous polystyrene (PS) foams demonstrated great potential for the effective absorption of organic solvents from underwater. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Development of germ-free plants and tissue culture

    NASA Technical Reports Server (NTRS)

    Venketeswaran, S.

    1973-01-01

    The botanical program is reported for experiments performed at the Lunar Receiving Laboratory. Papers prepared during this program are listed. The studies reported include: tissues cultured on various mediums, nutritional studies, preparation of plant cultures for Apollo 15, and pine tissue cultures.

  7. 78 FR 22922 - Submission for OMB Review; Comment Request

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-17

    ...-consuming applications annually, 4 applications of medium difficulty, and 10 of the least difficult... the exemptive order application process, including preparation and revision of an application and... the costs required to prepare other more complex and novel applications. See also Political...

  8. Controlled production of camembert-type cheeses: part III role of the ripening microflora on free fatty acid concentrations.

    PubMed

    Leclercq-Perlat, Marie-Noëlle; Corrieu, Georges; Spinnler, Henry-Eric

    2007-05-01

    Phenomena generating FFAs, important flavour precursors, are significant in cheese ripening. In Camembert-like cheeses, it was intended to establish the relationships between the dynamics of FFA concentrations changes and the succession of ripening microflora during ripening. Experimental Camembert-type cheeses were prepared in duplicate from pasteurised milk inoculated with Kluyveromyces lactis, Geotrichum candidum, Penicillium camemberti, and Brevibacterium aurantiacum under aseptic conditions. For each cheese and each cheesy medium, concentrations of FFAs with odd-numbered carbons, except for 9:0 and 13:0, did not change over time. For long-chain FFAs, concentrations varied with the given cheese part (rind or core). K. lactis produced only short or medium-chain FFAs during its growth and had a minor influence on caproic, caprylic, capric, and lauric acids in comparison with G. candidum, the most lipolytic of the strains used here. It generated all short or medium-chain FFAs (4:0-12:0) during its exponential and slowdown growth periods and only long-chain ones (14:0-18:0) during its stationary phase. Pen. camemberti produced more long-chain FFAs (14:0-18:0) during its sporulation. Brev. aurantiacum did not generate any FFAs. The evidence of links between specific FFAs and the growth of a given microorganism is shown.

  9. [Troubleshooting of bioinequivalence of compound valsartan tablets].

    PubMed

    Shao, Da; Zhang, Yi-Fan; Zhan, Yan; Chen, Xiao-Yan; Zhong, Da-Fang

    2014-04-01

    The study aims to evaluate the bioequivalence of valsartan hydrochlorothiazide tablets, and to investigate the potential cause of bioinequivalence. This was a single-center study with an open, randomized double-way crossover design. Test and reference preparations containing 160 mg of valsartan and 25 mg of hydrochlorothiazide were given to 36 healthy male volunteers. Plasma concentrations of valsartan and hydrochlorothiazide were determined simultaneously by LC-MS/MS. The pharmacokinetic parameters and relative bioavailability were calculated, while the bioequivalence between test and reference preparations were evaluated. The dissolution profiles of test and reference preparations in four different mediums were determined via dissolution test and HPLC. The similarity was investigated according to the similarity factors (f2). The F(o-t) and F(0-infinity) were (139.4 +/- 65.2)% and (137.5 +/- 61.2)% for valsartan of test preparations. It led to get the conclusion that test and reference preparations were not bioequivalent for valsartan. A significant difference was observed between test and reference tablets in the valsartan dissolution test of pH 1.2 hydrochloric acid solution. The key factor of the bioinequivalence might be that dissolution of valsartan in acid medium has marked difference between two preparations.

  10. Evaluation of cytotoxicity of different tobacco product preparations.

    PubMed

    Arimilli, Subhashini; Damratoski, Brad E; Bombick, Betsy; Borgerding, Michael F; Prasad, G L

    2012-12-01

    Acute exposure to cigarette smoke or its components triggers diverse cellular effects, including cytotoxicity. However, available data regarding the potential cytotoxic effects of smokeless tobacco (ST) extracts lack consensus. Here, we investigated the relative biological effects of 2S3 reference ST, and whether ST elicits differential cellular/molecular responses compared to combustible tobacco product preparations (TPPs) prepared from 3R4F cigarettes. Total particulate matter (TPM) and whole smoke conditioned medium (WS-CM) were employed as combustible TPPs, while the ST extract was used as non-combustible TPP. HL60, THP1 cells and human PBMCs were used to examine the effects of TPPs in short-term cell culture. Corresponding EC(50) values, normalized for nicotine content of the TPPs, suggest that combustible TPPs induced higher cytotoxicity as follows: WS-CM TPM ≥ ≫ST extract>nicotine. While all three TPPs induced detectable levels of DNA damage and IL8 secretion, the combustible TPPs were significantly more potent than the ST preparation. The major PBMC subsets showed differential cytotoxicity to combustible TPPs as follows: CD4>CD8>monocytes>NK cells. These findings suggest that, relative cytotoxic and other cell biological effects of TPPs are dose-dependent, and that ST extract is the least cytotoxic TPP tested in this study. Copyright © 2012 Elsevier Inc. All rights reserved.

  11. [Influence of antitumor preparations on the concentration of free radicals in cells of Fusarium bulbigenum var. blasticola fungus during primary and tumour-like secondary growth].

    PubMed

    Riabikin, Iu A; Nikitina, E T; Balgimbatva, A S; Zashkvara, O V; Shakiev, S Sh

    2007-01-01

    The fungus Fusarium bulbigenum var. blasticola in which secondary tumor-like formations appear under certain conditions in aging was used as a new test system to examine the action of antitumor preparations. Free radicals in the primary mycelium and tumor-like formations without introduction of preparations (control samples) and after the introduction of preparation into the cultivation medium of the fungus have been studied by EPR spectroscopy. The EPR spectra of the fungus represent single, somewhat asymmetrical lines with a width of deltaH = 0.4 divided by 0.6 mT and g = 2.0036 +/- 0.006, which enabled one to assign the paramagnetic centers observed to melanine radicals. It was found that the concentration of free radicals in tumor-like formations is always higher than in the primary mycelium, which may be related to intensive metabolism in tumor-like formations. It has been established that several antitumor preparations (fluorouracil, hydrea, methotrexat, and vepezide) completely inhibit the growth of tumor-like formations. Another group of preparations (cyclophosphanum, dacarbazin, adriablastin, and vinblastin), on the contrary, stimulate their growth, which is accompanied by an increase in the concentration of free radicals in cells of the primary mycelium and tumor-like formations. The preparations of the third group (mercaptopurine, lanvis, and farmorubicin), despite the increased level of free radicals in cells, have a weak inhibitory effect. It has been shown that, in the concentration range studied, vitamins B2, B12, C, and PP stimulate the growth of tumor-like formations, and, when used in combination with antitumor preparations, enhance or reduce the inhibitory properties of these preparations.

  12. An improved stress corrosion test medium for aluminum alloys

    NASA Technical Reports Server (NTRS)

    Humphries, T. S.; Coston, J. E.

    1981-01-01

    A laboratory test method that is only mildly corrosive to aluminum and discriminating for use in classifying the stress corrosion cracking resistance of aluminum alloys is presented along with the method used in evaluating the media selected for testing. The proposed medium is easier to prepare and less expensive than substitute ocean water.

  13. Using Structure, Action, and Power to Make Teacher Preparation Responsive to Public Law 94-142.

    ERIC Educational Resources Information Center

    Bryson, John M.

    Organizational design for change of teacher education institutions should focus on the shaping of political activities through forums, arenas, and courts. Forums provide the medium for expression and competition of alternate ideas in the institution, and arenas are the medium for achieving group objectives through cooperation, contest, or…

  14. Organic contaminant separator

    DOEpatents

    Del Mar, P.

    1993-12-28

    A process is presented of sample preparation prior to analysis for the concentration of an organic contaminant in an aqueous medium by (a) passing an initial aqueous medium including a minor amount of the organic contaminant through a composite tube comprised of a blend of a polyolefin and a polyester, the composite tube having an internal diameter of from about 0.1 to about 2.0 millimeters and being of sufficient length to permit the organic contaminant to adhere to the composite tube, (b) passing a solvent through the composite tube. The solvent is capable of separating the adhered organic contaminant from the composite tube. Further, an extraction apparatus is presented for sample preparation prior to analysis for the concentration of an organic contaminant in an aqueous medium. The apparatus includes a composite tube comprised of a blend of a polyolefin and a polyester. The composite tube has an internal diameter of from about 0.1 to about 2.0 millimeters and has sufficient length to permit an organic contaminant contained within an aqueous medium passed therethrough to adhere to the composite tube. 2 figures.

  15. Security writing application of thermal decomposition assisted NaYF4:Er3+/Yb3+ upconversion phosphor

    NASA Astrophysics Data System (ADS)

    Kumar, A.; Tiwari, S. P.; Esteves da Silva, Joaquim C. G.; Kumar, K.

    2018-07-01

    The authors have synthesized water-dispersible NaYF4:Er3+/Yb3+ upconversion particles via a thermal decomposition route and optimized the green upconversion emission through a concentration variation of the Yb3+ sensitizer. The prepared particles were found to be ellipsoid in shape having an average particle dimension of 600  ×  150 nm. It is observed that the sample with 18 mmol% Yb3+ ion concentration and 2 mmol% Er3+ ion gives optimum upconversion intensity in the green region under 980 nm excitation. Colloidal dispersibility of the sample in different solvents was checked and hexane was found to be the best medium for the prepared particles. The particle size of the sample was found to be suitable for the preparation of colloidal ink and security writing on a plain sheet of paper. This was demonstrated successfully using ink prepared in polyvinyl chloride gold medium.

  16. A new method of preparing embeddment-free sections for transmission electron microscopy: applications to the cytoskeletal framework and other three-dimensional networks

    PubMed Central

    1984-01-01

    Diethylene glycol distearate is used as a removable embedding medium to produce embeddment -free sections for transmission electron microscopy. The easily cut sections of this material float and form ribbons in a water-filled knife trough and exhibit interference colors that aid in the selection of sections of equal thickness. The images obtained with embeddment -free sections are compared with those from the more conventional epoxy-embedded sections, and illustrate that embedding medium can obscure important biological structures, especially protein filament networks. The embeddment -free section methodology is well suited for morphological studies of cytoskeletal preparations obtained by extraction of cells with nonionic detergent in cytoskeletal stabilizing medium. The embeddment -free section also serves to bridge the very different images afforded by embedded sections and unembedded whole mounts. PMID:6539336

  17. Simplified method for preparation of concentrated exoproteins produced by Staphylococcus aureus grown on surface of cellophane bag containing liquid medium.

    PubMed

    Ikigai, H; Seki, K; Nishihara, S; Masuda, S

    1988-01-01

    A simplified method for preparation of concentrated exoproteins including protein A and alpha-toxin produced by Staphylococcus aureus was successfully devised. The concentrated proteins were obtained by cultivating S. aureus organisms on the surface of a liquid medium-containing cellophane bag enclosed in a sterilized glass flask. With the same amount of medium, the total amount of proteins obtained by the method presented here was identical with that obtained by conventional liquid culture. The concentration of proteins obtained by the method, however, was high enough to observe their distinct bands stained on polyacrylamide gel electrophoresis. This method was considered quite useful not only for large-scale cultivation for the purification of staphylococcal proteins but also for small-scale study using the proteins. The precise description of the method was presented and its possible usefulness was discussed.

  18. Evidence of reversible bradycardia and arrhythmias caused by immunogenic proteins secreted by T. cruzi in isolated rat hearts.

    PubMed

    Rodríguez-Angulo, Héctor O; Toro-Mendoza, Jhoan; Marques, Juan A; Concepción, Juan L; Bonfante-Cabarcas, Rafael; Higuerey, Yoliver; Thomas, Luz E; Balzano-Nogueira, Leandro; López, José R; Mijares, Alfredo

    2015-02-01

    Chagas cardiomyopathy, caused by the protozoan Trypanosoma cruzi, is characterized by alterations in intracellular ion, heart failure and arrhythmias. Arrhythmias have been related to sudden death, even in asymptomatic patients, and their molecular mechanisms have not been fully elucidated. The aim of this study is to demonstrate the effect of proteins secreted by T. cruzi on healthy, isolated beating rat heart model under a non-damage-inducing protocol. We established a non-damage-inducing recirculation-reoxygenation model where ultrafiltrate fractions of conditioned medium control or conditioned infected medium were perfused at a standard flow rate and under partial oxygenation. Western blotting with chagasic patient serum was performed to determine the antigenicity of the conditioned infected medium fractions. We observed bradycardia, ventricular fibrillation and complete atrioventricular block in hearts during perfusion with >50 kDa conditioned infected culture medium. The preincubation of conditioned infected medium with chagasic serum abolished the bradycardia and arrhythmias. The proteins present in the conditioned infected culture medium of >50 kDa fractions were recognized by the chagasic patient sera associated with arrhythmias. These results suggest that proteins secreted by T. cruzi are involved in Chagas disease arrhythmias and may be a potential biomarker in chagasic patients.

  19. Evidence of Reversible Bradycardia and Arrhythmias Caused by Immunogenic Proteins Secreted by T. cruzi in Isolated Rat Hearts

    PubMed Central

    Rodríguez-Angulo, Héctor O.; Toro-Mendoza, Jhoan; Marques, Juan A.; Concepción, Juan L.; Bonfante-Cabarcas, Rafael; Higuerey, Yoliver; Thomas, Luz E.; Balzano-Nogueira, Leandro; López, José R.; Mijares, Alfredo

    2015-01-01

    Rationale Chagas cardiomyopathy, caused by the protozoan Trypanosoma cruzi, is characterized by alterations in intracellular ion, heart failure and arrhythmias. Arrhythmias have been related to sudden death, even in asymptomatic patients, and their molecular mechanisms have not been fully elucidated. Objective The aim of this study is to demonstrate the effect of proteins secreted by T. cruzi on healthy, isolated beating rat heart model under a non-damage-inducing protocol. Methods and Results We established a non-damage-inducing recirculation-reoxygenation model where ultrafiltrate fractions of conditioned medium control or conditioned infected medium were perfused at a standard flow rate and under partial oxygenation. Western blotting with chagasic patient serum was performed to determine the antigenicity of the conditioned infected medium fractions. We observed bradycardia, ventricular fibrillation and complete atrioventricular block in hearts during perfusion with >50 kDa conditioned infected culture medium. The preincubation of conditioned infected medium with chagasic serum abolished the bradycardia and arrhythmias. The proteins present in the conditioned infected culture medium of >50 kDa fractions were recognized by the chagasic patient sera associated with arrhythmias. Conclusions These results suggest that proteins secreted by T. cruzi are involved in Chagas disease arrhythmias and may be a potential biomarker in chagasic patients. PMID:25647069

  20. Impact of Nutrient Restriction on the Structure of Listeria monocytogenes Biofilm Grown in a Microfluidic System.

    PubMed

    Cherifi, Tamazight; Jacques, Mario; Quessy, Sylvain; Fravalo, Philippe

    2017-01-01

    Biofilm formation by the pathogen Listeria monocytogenes is a major concern in food industries. The aim of this work was to elucidate the effect of nutrient limitation on both biofilm architecture and on the viability of the bacteria in microfluidic growth conditions. Biofilm formation by two L. monocytogenes strains was performed in a rich medium (BHI) and in a 10-fold diluted BHI (BHI/10) at 30°C for 24 h by using both static conditions and the microfluidic system Bioflux. In dynamic conditions, biofilms grown in rich and poor medium showed significant differences as well in structure and in the resulting biovolume. In BHI/10, biofilm was organized in a knitted network where cells formed long chains, whereas in the rich medium, the observed structure was homogeneous cellular multilayers. Biofilm biovolume production in BHI/10 was significantly higher than in BHI in these dynamic conditions. Interestingly, biovolume of dead cells in biofilms formed under limited nutrient conditions (BHI/10) was significantly higher than in biofilms formed in the BHI medium. In the other hand, in static conditions, biofilm is organized in a multilayer cells and dispersed cells in a rich medium BHI and poor medium BHI/10 respectively. There was significantly more biomass in the rich medium compared to BHI/10 but no difference was noted in the dead/damaged subpopulation showing how L. monocytogenes biofilm could be affected by the growth conditions. This work demonstrated that nutrient concentration affects biofilm structure and the proportion of dead cells in biofilms under microfluidic condition. Our study also showed that limited nutrients play an important role in the structural stability of L. monocytogenes biofilm by enhancing cell death and liberating extracellular DNA.

  1. Impact of Nutrient Restriction on the Structure of Listeria monocytogenes Biofilm Grown in a Microfluidic System

    PubMed Central

    Cherifi, Tamazight; Jacques, Mario; Quessy, Sylvain; Fravalo, Philippe

    2017-01-01

    Biofilm formation by the pathogen Listeria monocytogenes is a major concern in food industries. The aim of this work was to elucidate the effect of nutrient limitation on both biofilm architecture and on the viability of the bacteria in microfluidic growth conditions. Biofilm formation by two L. monocytogenes strains was performed in a rich medium (BHI) and in a 10-fold diluted BHI (BHI/10) at 30°C for 24 h by using both static conditions and the microfluidic system Bioflux. In dynamic conditions, biofilms grown in rich and poor medium showed significant differences as well in structure and in the resulting biovolume. In BHI/10, biofilm was organized in a knitted network where cells formed long chains, whereas in the rich medium, the observed structure was homogeneous cellular multilayers. Biofilm biovolume production in BHI/10 was significantly higher than in BHI in these dynamic conditions. Interestingly, biovolume of dead cells in biofilms formed under limited nutrient conditions (BHI/10) was significantly higher than in biofilms formed in the BHI medium. In the other hand, in static conditions, biofilm is organized in a multilayer cells and dispersed cells in a rich medium BHI and poor medium BHI/10 respectively. There was significantly more biomass in the rich medium compared to BHI/10 but no difference was noted in the dead/damaged subpopulation showing how L. monocytogenes biofilm could be affected by the growth conditions. This work demonstrated that nutrient concentration affects biofilm structure and the proportion of dead cells in biofilms under microfluidic condition. Our study also showed that limited nutrients play an important role in the structural stability of L. monocytogenes biofilm by enhancing cell death and liberating extracellular DNA. PMID:28567031

  2. Pneumococcal whole-cell vaccine: optimization of cell growth of unencapsulated Streptococcus pneumoniae in bioreactor using animal-free medium.

    PubMed

    Liberman, C; Takagi, M; Cabrera-Crespo, J; Sbrogio-Almeida, M E; Dias, W O; Leite, L C C; Gonçalves, V M

    2008-11-01

    The high cost of the available pneumococcal conjugated vaccines has been an obstacle in implementing vaccination programs for children in developing countries. As an alternative, Malley et al. proposed a vaccine consisting of inactivated whole-cells of unencapsulated S. pneumoniae, which provides serotype-independent protection and involves lower production costs. Although the pneumococcus has been extensively studied, little research has focused on its large-scale culture, thus implying a lack of knowledge of process parameters, which in turn are essential for its successful industrial production. The strain Rx1Al- eryR was originally cultured in Todd-Hewitt medium (THY), which is normally used for pneumococcus isolation, but is unsuitable for human vaccine preparations. The purposes of this study were to compare the strains Rx1Al- eryR and kanR, develop a new medium, and generate new data parameters for scaling-up the process. In static flasks, cell densities were higher for eryR than kanR. In contrast, the optical density (OD) of the former decreased immediately after reaching the stationary phase, and the OD of the latter remained stable. The strain Rx1Al- kanR was cultivated in bioreactors with medium based on either acid-hydrolyzed casein (AHC) or enzymatically hydrolyzed soybean meal (EHS). Biomass production in EHS was 2.5 times higher than in AHC, and about ten times higher than in THY. The process developed for growing the strain Rx1Al- kanR in pH-controlled bioreactors was shown to be satisfactory to this fastidious bacterium. The new culture conditions using this animal-free medium may allow the production of the pneumococcal whole-cell vaccine.

  3. Evaluation of pathogenicity of Candida albicans in germination-ready states using a silkworm infection model.

    PubMed

    Matsumoto, Haruhito; Nagao, Jun-ichi; Cho, Tamaki; Kodama, Jun

    2013-01-01

    We previously developed an N-acetyl-D-glucosamine (GlcNAc) medium which induces Candida albicans to undergo a yeast-to-hyphal transition through a cAMP-PKA pathway. Microarray analysis demonstrated that 18 genes, including ALS3 that encodes a cell wall adhesion, were upregulated by 30-min incubation of yeast cells at 37°C in the GlcNAc medium. To investigate the differences between morphological transition and morphotype in C. albicans as a consequence of infection, this study utilized a silkworm infection model as an invertebrate mini-host. We prepared 3 different conditions of C. albicans cells in vitro by changing the incubation times in the GlcNAc medium: yeast-form cells at 0 min (Y0 cells), yeast-form cells in germination-ready state at 60 min (Y60 cells), and hyphal cells at 120 min (H120 cells), and compared their pathogenicities. We performed the infection study at various temperatures to find temperature-dependent virulence factors in vivo. Y60 cells in germination-ready state in the GlcNAc medium showed higher pathogenicity in vivo compared to Y0 and H120 cells at 30°C. Y60 cells proliferated in silkworms 24 h post-injection at 30°C, whereas the other 2 cell types did not. In vitro analysis demonstrated that Y60 cells, but not Y0 cells, germinated in the silkworm hemolymph at 30°C. However, Y0 and Y60 cells showed a similar degree of germination in the silkworm hemolymph at 37°C, although no significant difference in silkworm survival after infection with each cell type was observed at 37°C. These results suggested that the germination-ready state induced by the GlcNAc medium contributed to virulence in the silkworm.

  4. Mesenchymal stem cell-conditioned medium triggers neuroinflammation and reactive species generation in organotypic cultures of rat hippocampus.

    PubMed

    Horn, Ana Paula; Bernardi, Andressa; Luiz Frozza, Rudimar; Grudzinski, Patrícia Bencke; Hoppe, Juliana Bender; de Souza, Luiz Fernando; Chagastelles, Pedro; de Souza Wyse, Angela Terezinha; Bernard, Elena Aida; Battastini, Ana Maria Oliveira; Campos, Maria Martha; Lenz, Guido; Nardi, Nance Beyer; Salbego, Christianne

    2011-07-01

    Cell therapy using bone marrow-derived mesenchymal stem cells (MSCs) seems to be a new alternative for the treatment of neurodegenerative diseases. Despite several promising results with their use, possible side effects are still unknown. In a previous work, we have shown that MSC-conditioned medium is toxic to hippocampal slice cultures and aggravates cell death induced by oxygen and glucose deprivation. In this work, we investigated whether the inflammatory response and/or reactive species formation could be involved in that toxicity. Rat organotypic hippocampal cultures were exposed for 24 h to conditioned medium from MSCs isolated from rat bone marrow. A marked glial activation was observed after exposure of cultures to MSC-conditioned medium, as evidenced by glial fibrillary acid protein (GFAP) and isolectin B(4) increase. Tumor necrosis factor-α and interleukin-6 levels were increased in the culture medium, and 2,7-dihydrodichlorofluorescein diacetate oxidation (indicating reactive species generation) and inducible nitric oxide synthase (iNOS) immunocontent were also higher after exposure of cultures to MSC-conditioned medium. Antioxidants (ascorbic acid and TROLOX(®)), N(ω)-nitro-l-arginine methyl ester hydrochloride, and anti-inflammatory drugs (indomethacin and dexamethasone) reduced cell death in hippocampal organotypic cultures after their exposure to MSC-conditioned medium. The results obtained here suggest that MSC-secreted factors trigger reactive species generation and neuroinflammation in organotypic cultures of hippocampus, introducing a note of caution in the use of these cells for neurological application.

  5. Pharmacokinetic behavior of intravitreal triamcinolone acetonide prepared by a hospital pharmacy.

    PubMed

    Oishi, Masako; Maeda, Shinichiro; Hashida, Noriyasu; Ohguro, Nobuyuki; Tano, Yasuo; Kurokawa, Nobuo

    2008-01-01

    We developed a new hospital pharmaceutical preparation of triamcinolone acetonide (TA) for intravitreal injections using sodium hyaluronate as the vehicle. The purpose of this study was to compare the pharmacokinetic behavior of this hospital pharmacy preparation of TA (HPP-TA) to that of a commercial preparation of TA (CP-TA) in rats. We injected the two preparations of TA into the vitreous humor of male Wistar rats. The rats were killed between days 1 and 21, and the concentration of TA in the vitreous was measured by high-performance liquid chromatography to determine the pharmacokinetic parameters. We also examined the microscopic appearance of the TA particles in these preparations. The elimination half-life was 6.08 days for the CP-TA and 5.78 days for the HPP-TA. A two-compartment model was suitable to approximate the pharmacokinetic behavior of HPP-TA in the vitreous body, but this model was not suitable for CP-TA, because its pharmacokinetic behavior was not sufficiently stable. The particle size of CP-TA was largest, followed by TA powder and HPP-TA. Many particles were agglutinated in the CP-TA preparation, whereas the TA particles were fine and dispersed in the HPP-TA medium. The TA particle size and the suspension medium are likely important factors in the preparation of a safe and stable suspension of TA. HPP-TA satisfied these requirements and should be suitable for clinical use.

  6. Isolation of a hemidesmosome-rich fraction from a human squamous cell carcinoma cell line

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

    Hirako, Yoshiaki, E-mail: s47526a@cc.nagoya-u.ac.jp; Yonemoto, Yuki; Yamauchi, Tomoe

    2014-06-10

    Hemidesmosomes are cell-to-matrix adhesion complexes anchoring keratinocytes to basement membranes. For the first time, we present a method to prepare a fraction from human cultured cells that are highly enriched in hemidesmosomal proteins. Using DJM-1 cells derived from human squamous cell carcinoma, accumulation of hemidesmosomes was observed when these cells were cultured for more than 10 days in a commercial serum-free medium without supplemental calcium. Electron microscopy demonstrated that numerous electron-dense adhesion structures were present along the basal cell membranes of DJM-1 cells cultured under the aforementioned conditions. After removing cellular materials using an ammonia solution, hemidesmosomal proteins and depositedmore » extracellular matrix were collected and separated by electrophoresis. There were eight major polypeptides, which were determined to be plectin, BP230, BP180, integrin α6 and β4 subunits, and laminin-332 by immunoblotting and mass spectrometry. Therefore, we designated this preparation as a hemidesmosome-rich fraction. This fraction contained laminin-332 exclusively in its unprocessed form, which may account for the promotion of laminin deposition, and minimal amounts of Lutheran blood group protein, a nonhemidesmosomal transmembrane protein. This hemidesmosome-rich fraction would be useful not only for biological research on hemidesmosomes but also for developing a serum test for patients with blistering skin diseases. - Highlights: • A defined condition promoted accumulation of hemidesmosomes in human cultured cells. • A fraction isolated from the cells contained eight major polypeptides. • The polypeptides were the five major hemidesmosome proteins and laminin-332. • The cultured cells deposited laminin-332 in its unprocessed form under the condition. • We report a method to prepare a fraction highly enriched in hemidesmosome proteins.« less

  7. Quenching of porous silicon photoluminescence by molecular oxygen and dependence of this phenomenon on storing media and method of preparation of pSi photosensitizer

    NASA Astrophysics Data System (ADS)

    Balaguer, María; Matveeva, Eugenia

    2010-10-01

    The quenching of porous silicon photoluminescence (pSi PL) by molecular oxygen has been studied in different storing media in an attempt to clarify the mechanism of the energy transfer from the silicon photosensitizer to the oxygen acceptor. Luminescent materials have been prepared by two methods: electrochemical anodizing and chemical etching. Different structural forms were used: porous layers on silicon wafer and two kinds of differently prepared powder. Dry air and liquid water were employed as storing media; quenching behaviour was under observation until total degradation of quenching properties. Singlet oxygen molecules generation through energy transfer from photoluminescent pSi was the only photosensitizing mechanism observed under dry gas conditions. This PL quenching process was preferentially developed at 760 nm (1.63 eV) that corresponds to the formation of the 1Σ singlet oxygen state. Oxidation of the pSi photosensitizer was the main factor that led to its total deactivation in a time scale of few weeks. Regarding water medium, different photosensitizing behaviour was observed. In watery conditions, two preferred energy levels were found: the one detected in dry gas and another centred at approximately 2.2 eV (550 nm). Formation of reactive oxygen species (ROS) different from singlet oxygen, such as superoxide anion or superoxide radical, can be responsible for the second one. This second quenching process developed gradually after the initial contact of pSi photosensitizer with water and then degraded. The process lasted only several hours. Therefore, functionalization of the pSi photosensitizer is probably required to stabilize its PL and quenching properties in the watery physiological conditions required for biomedical applications.

  8. Nanocrystalline hydroxyapatite ceramics prepared by hydrolysis in polyol medium

    NASA Astrophysics Data System (ADS)

    Mechay, Abderrahmen; Feki, Hafed E. L.; Schoenstein, Fréderic; Jouini, Noureddine

    2012-07-01

    This Letter describes a new approach for the synthesis of hydroxyapatite nanoparticles, which involves precipitation and hydrolysis reactions conducted in polyol medium. In fact, ammonium-hydrogen phosphate and calcium nitrate were dissolved in polyol, and then heated at the boiling point of the polyol (ethane1, 2diol or propane1, 2diol). Besides, the phase and composition of the polycrystalline were studied by TGA/DTA, FT-IR, TEM and XRD techniques. The nanoparticles thus obtained present interesting morphological characters varying from needle to very thin platelet. Moreover, the hydroxyapatite prepared in ployol shows higher cristallinity in comparison with that obtained by other 'chimie douce' methods.

  9. [Clinical practice guidelines in Peru: evaluation of its quality using the AGREE II instrument].

    PubMed

    Canelo-Aybar, Carlos; Balbin, Graciela; Perez-Gomez, Ángela; Florez, Iván D

    2016-01-01

    To evaluate the methodological quality of clinical practice guidelines (CPGs) put into practice by the Peruvian Ministry of Health (MINSA), 17 CPGs from the ministry, published between 2009 and 2014, were independently evaluated by three methodologic experts using the AGREE II instrument. The score of AGREE II domains was low and very low in all CPGs: scope and purpose (medium, 44%), clarity of presentation (medium, 47%), participation of decision-makers (medium, 8%), methodological rigor (medium, 5%), applicability (medium, 5%), and editorial independence (medium, 8%). In conclusion, the methodological quality of CPGs implemented by the MINSA is low. Consequently, its use could not be recommended. The implementation of the methodology for the development of CPGs described in the recentlypublished CPG methodological preparation manual in Peru is a pressing need.

  10. A novel sample preparation method to avoid influence of embedding medium during nano-indentation

    NASA Astrophysics Data System (ADS)

    Meng, Yujie; Wang, Siqun; Cai, Zhiyong; Young, Timothy M.; Du, Guanben; Li, Yanjun

    2013-02-01

    The effect of the embedding medium on the nano-indentation measurements of lignocellulosic materials was investigated experimentally using nano-indentation. Both the reduced elastic modulus and the hardness of non-embedded cell walls were found to be lower than those of the embedded samples, proving that the embedding medium used for specimen preparation on cellulosic material during nano-indentation can modify cell-wall properties. This leads to structural and chemical changes in the cell-wall constituents, changes that may significantly alter the material properties. Further investigation was carried out to detect the influence of different vacuum times on the cell-wall mechanical properties during the embedding procedure. Interpretation of the statistical analysis revealed no linear relationships between vacuum time and the mechanical properties of cell walls. The quantitative measurements confirm that low-viscosity resin has a rapid penetration rate early in the curing process. Finally, a novel sample preparation method aimed at preventing resin diffusion into lignocellulosic cell walls was developed using a plastic film to wrap the sample before embedding. This method proved to be accessible and straightforward for many kinds of lignocellulosic material, but is especially suitable for small, soft samples.

  11. Development and validation of spectrophotometric methods for estimating amisulpride in pharmaceutical preparations.

    PubMed

    Sharma, Sangita; Neog, Madhurjya; Prajapati, Vipul; Patel, Hiren; Dabhi, Dipti

    2010-01-01

    Five simple, sensitive, accurate and rapid visible spectrophotometric methods (A, B, C, D and E) have been developed for estimating Amisulpride in pharmaceutical preparations. These are based on the diazotization of Amisulpride with sodium nitrite and hydrochloric acid, followed by coupling with N-(1-naphthyl)ethylenediamine dihydrochloride (Method A), diphenylamine (Method B), beta-naphthol in an alkaline medium (Method C), resorcinol in an alkaline medium (Method D) and chromotropic acid in an alkaline medium (Method E) to form a colored chromogen. The absorption maxima, lambda(max), are at 523 nm for Method A, 382 and 490 nm for Method B, 527 nm for Method C, 521 nm for Method D and 486 nm for Method E. Beer's law was obeyed in the concentration range of 2.5-12.5 microg mL(-1) in Method A, 5-25 and 10-50 microg mL(-1) in Method B, 4-20 microg mL(-1) in Method C, 2.5-12.5 microg mL(-1) in Method D and 5-15 microg mL(-1) in Method E. The results obtained for the proposed methods are in good agreement with labeled amounts, when marketed pharmaceutical preparations were analyzed.

  12. 40 CFR 205.57-3 - Test vehicle preparation.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Test vehicle preparation. 205.57-3 Section 205.57-3 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) NOISE ABATEMENT PROGRAMS TRANSPORTATION EQUIPMENT NOISE EMISSION CONTROLS Medium and Heavy Trucks § 205.57-3 Test vehicle...

  13. Evaluation of Escherichia coli O157:H7 growth media for use in test-and-hold procedures for ground beef processing.

    PubMed

    Guerini, Michael N; Arthur, Terrance M; Shackelford, Steven D; Koohmaraie, Mohammad

    2006-05-01

    Since the mid-1990s, the beef industry has used a process called test and hold, wherein beef trim and ground beef are tested to keep products contaminated with Escherichia coli O157:H7 out of commerce. Current O157:H7 detection methods rely on a threshold level of bacterial growth for detection, which is dependent on the growth medium used. Twelve media were examined for growth and doubling time: buffered peptone water (BPW), SOC (which contains tryptone, yeast extract, KCl, MgCl2, and glucose), buffered peptone water plus SOC (BPW-SOC), Bacto-NZYM, RapidChek E. coli O157:H7 medium, BioControl EHEC8 culture medium, Neogen Reveal for E. coli O157:H7--Eight Hour medium (Neogen Reveal 8), BAX System medium for E. coli O157:H7 (BAX) BAX System medium for E. coli O157:H7 MP (BAX-MP), modified E. coli broth, nutrient medium, and tryptic soy broth (TSB). All media were tested at 37 or 42 degrees C under static or shaking conditions. The eight media with the highest total CFU per milliliter and most rapid doubling times were BPW-SOC, NZYM, RapidChek, EHEC8, Neogen Reveal 8, BAX, BAX-MP, and TSB. The ability of these eight media to enrich E. coli O157:H7 in ground beef was further evaluated through time-course experiments using immunomagnetic separation. Of these media, TSB was the easiest to prepare, had a wide application base, and was the least expensive. In the test-and-hold process, the normal ratio of medium to product is 1:10. In this study, a 1:3 ratio worked as well as a 1:10 ratio. Processors using test-and-hold procedures could use 1 liter of TSB to enrich for E. coli O157:H7 in a 375-g sample instead of the usual 3.375 liters, thus saving reagents, time, and labor while maintaining accuracy.

  14. A facile sol-gel synthesis of impurity-free nanocrystalline titania.

    PubMed

    Vinogradov, Alexandr V; Ermakova, Al'ena V; Butman, Mikhail F; Hey-Hawkins, Evamarie; Vinogradov, Vladimir V

    2014-06-14

    This paper reports an original technique that provides a highly pure crystalline sol of titania with controllable particle size by ultrasonic activation of the hydrolysis products of titanium isopropoxide in an aqueous medium at a near-neutral pH, which is potentially promising in impurity-sensitive electronics and biochemical engineering. Optimal conditions (H2O/TIP ratio, sonication time, etc.) for preparation of stable nanocrystalline titania sol were adopted. A new mechanism of regulation of aggregation and polycondensation under ultrasonic irradiation is proposed. Entrapment of human serum albumin (HSA) in the formed porous titania matrix results in high thermal stability of the protein dopants: the denaturation temperature of HSA is shifted by 31 °C.

  15. A novel organic solvent-based coupling method for the preparation of covalently immobilized proteins on gold.

    PubMed Central

    Parker, M. C.; Patel, N.; Davies, M. C.; Roberts, C. J.; Tendler, S. J.; Williams, P. M.

    1996-01-01

    A novel organic solvent-based coupling method has been developed for the covalent immobilization of biological material to gold surfaces. The method employs the polar organic solvent anhydrous 2,2,2-trifluoroethanol as the reaction medium and involves dissolution of the protein (catalase) in the solvent allowing protein coupling to proceed under basic conditions in a dry organic environment. The advantage of this method is that protein attachment is favored over hydrolysis of the coupling reagent. We have shown qualitatively and quantitatively that following attachment to the gold surface a significant proportion of the enzyme catalase remains catalytically active (at least 20-31%). PMID:8931151

  16. Growth factors for nanobacteria

    NASA Astrophysics Data System (ADS)

    Ciftcioglu, Neva; Kajander, E. Olavi

    1999-12-01

    Nanobacteria are novel microorganisms recently isolated from fetal bovine serum and blood of cows and humans. These coccoid, gram negative bacteria in alpha-2 subgroup of Proteobacteria grow slowly under mammalian cell culture conditions but not in common media for microbes. Now we have found two different kinds of culture supplement preparations that improve their growth and make them culturable in the classical sense. These are supernatant fractions of conditioned media obtained from 1 - 3 months old nanobacteria cultures and from about a 2 weeks old Bacillus species culture. Both improved multiplication and particle yields and the latter increased their resistance to gentamicin. Nanobacteria cultured with any of the methods shared similar immunological property, structure and protein pattern. The growth supporting factors were heat-stabile and nondialyzable, and dialysis improved the growth promoting action. Nanobacteria formed stony colonies in a bacteriological medium supplemented with the growth factors. This is an implication that nanobacterial growth is influenced by pre-existing bacterial flora.

  17. Enhancing learning using questions, adjunct to science charts

    NASA Astrophysics Data System (ADS)

    Holliday, William G.; Benson, Garth

    This study supported two hypotheses. First, adjunct questions interacted with a science chart so powerfully that content established as difficult to learn in the pilot and in this study's control groups became easier to learn when charted. Second, students familiar with the chart test before instruction (test exposure) were better prepared to take this test after instruction. This adjunct-question study examined the generalizability of selective-attention and academic-studying hypotheses to a modified science chart medium. About 300 high school students were randomly assigned to four conditions each including a vitamin chart (chart only, test exposure, importance of questions emphasized to students by teachers, and combinational conditions - test exposure and question importance) across 16 biology classrooms. Then these same students were again randomly assigned within each classroom to a control and to four question treatments no questions, questions focusing on easy-to-learn charted content, questions focusing on difficult-to-learn charted content, and a combinational treatment.

  18. Enhancing learning using questions adjunct to science charts

    NASA Astrophysics Data System (ADS)

    Holliday, William G.; Benson, Garth

    This study supported two hypotheses. First, adjunct questions interacted with a science chart so powerfully that content established as difficult to learn in the pilot and in this study's control groups became easier to learn when charted. Second, students familiar with the chart test before instruction (test exposure) were better prepared to take this test after instruction. This adjunct-question study examined the generalizability of selective-attention and academic-studying hypotheses to a modified science chart medium. About 300 high school students were randomly assigned to four conditions each including a vitamin chart (chart only, test exposure, importance of questions emphasized to students by teachers, and combinational conditions--test exposure and question importance) across 16 biology classrooms. Then these same students were again randomly assigned within each classroom to a control and to four question treatments (no questions, questions focusing on easy-to-learn charted content, questions focusing on difficult-to-learn charted content, and a combinational treatment).

  19. Detection of fibrils associated with Rickettsia rickettsii.

    PubMed

    Todd, W J; Burgdorfer, W; Wray, G P

    1983-09-01

    The ultrastructural appearance of the "halozone" formed at the interface between the spotted fever agent Rickettsia rickettsii and the cytoplasm of persistently infected cultured vole cells (Microtus pennsylvanicus) was studied by transmission electron microscopy. In sections of epoxy-embedded specimens stained with uranyl acetate and lead citrate, the halozone appeared clear and devoid of ultrastructural features. However, when unembedded preparations of whole infected cells were examined at 1,000 kV, fine structural features were observed within the halozone. These features, associated with the rickettsial outer membrane, were more clearly detectable when the infected cells were extracted with the detergent Triton X-100 before fixation. Under such conditions, long extensions of the rickettsial outer membrane, microfilament-like structures attached to that membrane, and extensive attachments between adjacent rickettsiae were seen. The fine structural features within the rickettsial halozone were also seen at 75 kV when unembedded sections were prepared from polyethylene glycol-embedded specimens. Thus, epoxy-embedding medium obscures the fine structural features within the halozone surrounding the rickettsiae in infected cells.

  20. Detection of fibrils associated with Rickettsia rickettsii.

    PubMed Central

    Todd, W J; Burgdorfer, W; Wray, G P

    1983-01-01

    The ultrastructural appearance of the "halozone" formed at the interface between the spotted fever agent Rickettsia rickettsii and the cytoplasm of persistently infected cultured vole cells (Microtus pennsylvanicus) was studied by transmission electron microscopy. In sections of epoxy-embedded specimens stained with uranyl acetate and lead citrate, the halozone appeared clear and devoid of ultrastructural features. However, when unembedded preparations of whole infected cells were examined at 1,000 kV, fine structural features were observed within the halozone. These features, associated with the rickettsial outer membrane, were more clearly detectable when the infected cells were extracted with the detergent Triton X-100 before fixation. Under such conditions, long extensions of the rickettsial outer membrane, microfilament-like structures attached to that membrane, and extensive attachments between adjacent rickettsiae were seen. The fine structural features within the rickettsial halozone were also seen at 75 kV when unembedded sections were prepared from polyethylene glycol-embedded specimens. Thus, epoxy-embedding medium obscures the fine structural features within the halozone surrounding the rickettsiae in infected cells. Images PMID:6411620

  1. Antimicrobial cellulosic hydrogel from olive oil industrial residue.

    PubMed

    Dacrory, Sawsan; Abou-Yousef, Hussein; Abouzeid, Ragab E; Kamel, Samir; Abdel-Aziz, Mohamed S; El-Badry, Mohamed

    2018-05-25

    The cellulose-based antimicrobial hydrogel was prepared from seed and husk cellulosic fibers of olive industry residues by load silver nanoparticles (AgNPs) onto grafted acrylamide monomer (Am) cellulosic fibers. The grafting approach was the free radical mechanism by utilizing ceric ammonium nitrate (CAN) as initiator in aqueous medium and N,N methylene bisacrylamide (MBAm) as a cross linker. The effect of different grafting conditions on the properties of produced hydrogels has been studied by determining the grafting parameters, i.e. concentration of Am, MBAm, grafting time and temperature to optimize grafting yield (G %), grafting efficiency (GE %), and swelling %. Characterizations of the obtained hydrogels were performed through monitoring swelling behavior, FTIR spectroscopy, SEM, and EDX. AgNPs were grown into the prepared hydrogel. Hydrogel/AgNPs were characterized by FT-IR spectroscopy, X-ray diffraction (XRD) and scanning electron microscopy (SEM). The hydrogel loaded AgNPs exhibit high efficient antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. Copyright © 2018. Published by Elsevier B.V.

  2. Corrosion Protection Of Mild Steel In Sea Water Using Chemical Inhibitor

    NASA Astrophysics Data System (ADS)

    Araoyinbo, Alaba O.; Salleh, Mohd Arif Anuar Mohd; Zulerwan Jusof, Muhammad

    2018-03-01

    The effect of sodium nitrite as a corrosion inhibitor of mild steel in sea water (i.e ASTM standard prepared sea water and sea water obtained from a local river) was investigated, using the weight loss technique. Different amount of sodium nitrite were prepared (i.e 2 % to 10 %) in the inhibition of the mild steel corrosion in sea water exposed to irradiation condition from sunlight exposure. The cut samples of mild steel were exposed to these corrosive media and the corresponding weight loss subsequently obtained was recorded at intervals of 1 to 4 weeks. It was observed that corrosion rate increases with the time of exposure to the corrosive medium exposed to sunlight and that sodium nitrite that was used at the chemical inhibitor was able to retard the corrosion rate of mild steel if the appropriate concentration is applied. The results obtained from the weight loss analysis shows that the optimum percentage of sodium nitrate in sea water that gives the optimum corrosion inhibition of mild steel is 4 %.

  3. Organic-inorganic hybrid carbon dots for cell imaging

    NASA Astrophysics Data System (ADS)

    Liu, Huan; Zhang, Hongwen; Li, Jiayu; Tang, Yuying; Cao, Yu; Jiang, Yan

    2018-04-01

    In this paper, nitrogen-doped carbon dots (CDs) had been synthesized directly by one-step ultrasonic treatment under mild conditions. During the functionalization process, Octa-aminopropyl polyhedral oligomeric silsesquioxane hydrochloride salt (OA-POSS) was used as stabilizing and passivation agent, which lead to self-assembling of CDs in aqueous medium solution. OA-POSS was obtained via hydrolytic condensation of γ-aminopropyl triethoxy silane (APTES). The average size of CDs prepared was approximately 3.3 nm with distribution between 2.5 nm and 4.5 nm. The prepared organic-inorganic hybrid carbon dots have several characteristics such as photoluminescence emission wavelength, efficient cellular uptake, and good biocompatibility. The results indicate that OA-POSS can maintain the fluorescence properties of the carbon dots effectively, and reduced cytotoxicity provides the possibility for biomedical applications. More than 89% of the Hela cells were viable when incubated with 2 mg ml‑1 or lesser organic-inorganic hybrid carbon dots. Thus, it provides a potential for multicolor imaging with HeLa cells.

  4. A strategy for the preparation of thioantimonates based on the concept of weak acids and corresponding strong bases.

    PubMed

    Anderer, Carolin; Delwa de Alarcón, Natalie; Näther, Christian; Bensch, Wolfgang

    2014-12-15

    By following a new synthetic approach, which is based on the in situ formation of a basic medium by the reaction between the strong base Sb(V)S4 (3-) and the weak acid H2 O, it was possible to prepare three layered thioantimonate(III) compounds of composition [TM(2,2'-bipyridine)3 ][Sb6 S10 ] (TM=Ni, Fe) and [Ni(4,4'-dimethyl-2,2'-bipyridine)3 ][Sb6 S10 ] under hydrothermal conditions featuring two different thioantimonate(III) network topologies. The antimony source, Na3 SbS4 ⋅ 9 H2 O, undergoes several decomposition reactions and produces the Sb(III) S3 species, which condenses to generate the layered anion. The application of transition-metal complexes avoids crystallization of dense phases. The reactions are very fast compared to conventional hydrothermal/solvothermal syntheses and are much less sensitive to changes of the reaction parameters. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Biosynthesis of Se-methyl-seleno-l-cysteine in Basidiomycetes fungus Lentinula edodes (Berk.) Pegler.

    PubMed

    Klimaszewska, M; Górska, S; Dawidowski, M; Podsadni, P; Turło, J

    2016-01-01

    The aim of the current study was to investigate whether the Basidiomycetes fungus Lentinula edodes can biosynthesize Se-methyl-seleno-l-cysteine, a seleno-amino acid with strong anticancer activity, and to optimize the culture conditions for its biosynthesis. We hypothesize that preparations obtained from Se-methyl-seleno-l-cysteine-enriched mycelia from this medicinal mushroom would possess stronger cancer-preventive properties than current preparations. By optimizing the concentration of selenium in the culture medium, we increased the mycelial concentration of Se-methyl-seleno-l-cysteine from essentially non-detectable levels to 120 µg/g dry weight. Significantly elevated levels of this amino acid also correlated with significant (twofold) inhibition of mycelial growth. Increases in the concentration of mycelial Se-methyl-seleno-l-cysteine appeared to be highly correlated with the enhanced biosynthesis of selenomethionine and total selenium content in mycelium. We have demonstrated that in L. edodes, enhanced biosynthesis of this non-protein amino acid eliminates excess selenium.

  6. Cellular uptake and transport of zein nanoparticles: effects of sodium caseinate.

    PubMed

    Luo, Yangchao; Teng, Zi; Wang, Thomas T Y; Wang, Qin

    2013-08-07

    Cellular evaluation of zein nanoparticles has not been studied systematically due to their poor redispersibility. Caseinate (CAS)-stabilized zein nanoparticles have been recently developed with better redispersibility in salt solutions. In this study, zein-CAS nanoparticles were prepared with different zein/CAS mass ratios. The prepared nanoparticles demonstrated good stabilities to maintain particle size (120-140 nm) in cell culture medium and HBSS buffer at 37 °C. The nanoparticles showed no cytotoxicity for Caco-2 cells for 72 h. CAS not only significantly enhanced cell uptake of zein nanoparticles in a concentration- and time-dependent manner but also remarkably improved epithelial transport through Caco-2 cell monolayer. The cell uptake of zein-CAS nanoparticles indicated an energy-dependent endocytosis process as evidenced by cell uptake under blocking conditions, that is, 4 °C, sodium azide, and colchicine. Fluorescent microscopy clearly showed the internalization of zein-CAS nanoparticles. This study may shed some light on the cellular evaluations of hydrophobic protein nanoparticles.

  7. Preparation of gold nanoparticle aggregates and their photothermal heating property.

    PubMed

    Kim, Jun-Hyun; Lavin, Brian W

    2011-01-01

    This report describes simple synthetic strategies to prepare partially aggregated gold nanoparticles (GNPs) and their ability to produce photothermally-induced heating of an aqueous medium upon exposure to broadband light. The formation of various GNPs and their aggregates were accomplished in the absence of surfactants at room temperature. The morphologies, structures, and absorption properties of these GNPs were carefully characterized. Given that the resulting GNPs possessing strong and wide absorption bands fall in the most intense solar radiation spectrum, the photothermally-induced heating of water was examined in the presence of the GNPs via irradiation with a solar simulator (i.e., 100 mW/cm2; 1-sun condition). Our GNPs exhibited a slightly greater increase in the water temperature (3-4 degrees C) than that of conventional citrate-stabilized GNPs. This superior photothermal heating property of our GNPs directly indicated that the intense and broad absorption band effectively improved the conversion of highly absorbed photon energy into heat.

  8. Design and environmentally benign synthesis of novel thiophene appended pyrazole analogues as anti-inflammatory and radical scavenging agents: Crystallographic, in silico modeling, docking and SAR characterization.

    PubMed

    Prabhudeva, Malledevarapura Gurumurthy; Bharath, Srinivasan; Kumar, Achutha Dileep; Naveen, Shivalingegowda; Lokanath, Neratur Krishnappagowda; Mylarappa, Bantaganahalli Ningappa; Kumar, Kariyappa Ajay

    2017-08-01

    Oxidative-stress induces inflammatory diseases and infections caused by drug-resistant microbial strains are on the rise necessitating the discovery of novel small-molecules for intervention therapy. The current study presents an effective and new green protocol for the synthesis of thiophene-appended pyrazoles through 3+2 annulations method. Chalcones 3(a-g) were prepared from 5-chloro-2-acetylthiophene and aromatic aldehydes by Claisen-Schmidt approach. The reaction of chalcones 3(a-g) with phenylhydrazine hydrochlorides 4(a-b) in acetic acid (30%) medium and also with freshly prepared citrus extract medium under reflux conditions produced the thiophene appended pyrazoles 5(a-l) in moderate yields. Structures of synthesized new pyrazoles were confirmed by spectral studies, elemental analysis and single crystal X-ray diffraction studies. Further, preliminary assessment of the anti-inflammatory properties of the compounds showed that, amongst the series, compounds 5d, 5e and 5l have excellent anti-inflammatory activities. Further, compounds 5c, 5d, 5g, and 5i exhibited excellent DPPH radical scavenging abilities in comparison with the standard ascorbic acid. Furthermore, using detailed structural modeling and docking efforts, combined with preliminary SAR, we show possible structural and chemical features on both the small-molecules and the protein that might contribute to the binding and inhibition. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. On the possibility of nonfat frying using molten glucose.

    PubMed

    Al-Khusaibi, Mohammed; Ahmad Tarmizi, Azmil Haizam; Niranjan, Keshavan

    2015-01-01

    Fried products impose a health concerns due to considerable amount of oil they contain. Production of snack foods with minimal oil content and good management of oil during frying to minimize the production of toxic compounds continue to be challenging aims. This paper aims to investigate the possibility of producing a fat-free food snack by replacing frying oil with a nonfat medium. Glucose was melted and its temperature was then brought to 185 °C and used to fry potato strips, to obtain a product referred here as glucose fries. The resulting product was compared with French fries prepared conventionally under conditions that resulted in similar final moisture content. The resulting products were also examined for crust formation, texture parameters, color development and glucose content. Stereo microscope images showed that similar crusts were formed in the glucose fries and French fries. Texture parameters were found to be similar for both products at 5 and 2 mm penetration depth. The maximum hardness at 2 mm penetration depth was also similar for both products, but different from cooked potato. The color development that characterized French fries was also observed in glucose fries. The glucose content in glucose fries was found to be twice the content of French fries, which is to be expected because glucose absorbed or adhered to the surface. In conclusion, glucose fries, with similar texture and color characteristics to that of French fries, can be prepared by using a nonfat frying medium. © 2014 Institute of Food Technologists®

  10. High cell density cultivation of a recombinant Escherichia coli strain expressing a 6-O-sulfotransferase for the production of bioengineered heparin.

    PubMed

    Zhang, J; Suflita, M; Fiaschetti, C M; Li, G; Li, L; Zhang, F; Dordick, J S; Linhardt, R J

    2015-01-01

    One of six heparin biosynthetic enzymes, cloned and expressed in Escherichia coli as a soluble fusion protein, requires large-scale preparation for use in the chemoenzymatic synthesis of heparin, an important anticoagulant drug. The 6-O-sulfotransferase isoform-3 (6-OST-3) can be conveniently prepared at mg/L levels in the laboratory by culturing E. coli on Luria-Bertani medium in shake flasks and inducing with isopropyl β-D-1-thiogalactopyranoside at an optical density of 0·6-0·8. The production of larger amounts of 6-OST-3 required fed-batch cultivation of E. coli in a stirred tank fermenter on medium containing an inexpensive carbon source, such as glucose or glycerol. The cultivation of E. coli on various carbon sources under different feeding schedules and induction strategies was examined. Conditions were established giving yields (5-20 mg g-cell-dry weight(-1)) of active 6-OST-3 with excellent productivity (2-5 mg l(-1) h(-1)). The production of 6-OST-3 in a fed-batch fermentation on an inexpensive carbon source has been demonstrated. The ability to scale-up the production of heparin biosynthetic enzymes, such as 6-OST-3, is critical for scaling-up the chemoenzymatic synthesis of heparin. The success of this project may someday lead to a commercially viable bioengineered heparin to replace the animal-sourced anticoagulant product currently on the market. © 2014 The Society for Applied Microbiology.

  11. Nano cube of CaSnO3: Facile and green co-precipitation synthesis, characterization and photocatalytic degradation of dye

    NASA Astrophysics Data System (ADS)

    Moshtaghi, Saeed; Gholamrezaei, Sousan; Salavati Niasari, Masoud

    2017-04-01

    In this work, nanocubes of CaSnO3 have been prepared by a simple and green co-precipitation method. In this technique, for preparation of calcium stannate, we have used from a complex structure of calcium as a new precursor and the synthesis of CaSnO3 have been done in water as a green solvent. Using of complexing precursors were created a congestion in reaction medium. Different conditions have been studied in synthetic approaches and optimized the effect of different parameters on the morphology of product such as precipitation agent (alkaline), pH, temperature, the rate of stirrer, surfactants and the mole ratio of surfactants for preparation product and obtain the best product in terms of quality and morphology. By using of this CaSnO3, two types of azo dyes (acid blue 92 and acid brown 14) have been degraded at presence of ultraviolet light from aqueous solution. Results display that the powder shows appropriate catalytic behavior for degradation of dyes (77% acid brown 14 and 67% acid black 92). Therefore these nano-cube structures have been used as photocatalysts in presence of UV light for degradation of azo dyes.

  12. An improved medium for the anaerobic growth of Paracoccus denitrificans Pd1222

    PubMed Central

    Hahnke, Stefanie M.; Moosmann, Philipp; Erb, Tobias J.; Strous, Marc

    2014-01-01

    Paracoccus denitrificans is a well studied model organism with respect to its aerobic and anaerobic respiratory enzymes. However, until now, the growth medium for this organism has not been optimized for anaerobic growth. In particular, the requirements of P. denitrificans for trace elements (TEs) are not well known. In the present study we aimed to improve growth rates of P. denitrificans Pd1222 on a defined medium under anoxic conditions. We designed media containing different combinations of TEs at various concentrations, and tested their performance against previously reported media. Our results suggest that growth rate and yield depend on the availability and concentration of TEs in the medium. A chelated TE solution was more suitable than an acidified TE solution. Highest growth rates were achieved with medium comprising the TEs iron, manganese, molybdenum, copper and zinc ranging from 0.1 to 9 μM. On this medium, P. denitrificans Pd1222 grew with a generation time of 4.4 h under anoxic conditions and 2.8 h under oxic conditions. Diauxic growth was clearly shown with respect to nitrate and nitrite reduction under anoxic conditions. PMID:24550891

  13. An improved medium for the anaerobic growth of Paracoccus denitrificans Pd1222.

    PubMed

    Hahnke, Stefanie M; Moosmann, Philipp; Erb, Tobias J; Strous, Marc

    2014-01-01

    Paracoccus denitrificans is a well studied model organism with respect to its aerobic and anaerobic respiratory enzymes. However, until now, the growth medium for this organism has not been optimized for anaerobic growth. In particular, the requirements of P. denitrificans for trace elements (TEs) are not well known. In the present study we aimed to improve growth rates of P. denitrificans Pd1222 on a defined medium under anoxic conditions. We designed media containing different combinations of TEs at various concentrations, and tested their performance against previously reported media. Our results suggest that growth rate and yield depend on the availability and concentration of TEs in the medium. A chelated TE solution was more suitable than an acidified TE solution. Highest growth rates were achieved with medium comprising the TEs iron, manganese, molybdenum, copper and zinc ranging from 0.1 to 9 μM. On this medium, P. denitrificans Pd1222 grew with a generation time of 4.4 h under anoxic conditions and 2.8 h under oxic conditions. Diauxic growth was clearly shown with respect to nitrate and nitrite reduction under anoxic conditions.

  14. The Role of Paracrine and Autocrine Signaling in the Early Phase of Adipogenic Differentiation of Adipose-derived Stem Cells

    PubMed Central

    Hemmingsen, Mette; Vedel, Søren; Skafte-Pedersen, Peder; Sabourin, David; Collas, Philippe; Bruus, Henrik; Dufva, Martin

    2013-01-01

    Introduction High cell density is known to enhance adipogenic differentiation of mesenchymal stem cells, suggesting secretion of signaling factors or cell-contact-mediated signaling. By employing microfluidic biochip technology, we have been able to separate these two processes and study the secretion pathways. Methods and results Adipogenic differentiation of human adipose-derived stem cells (ASCs) cultured in a microfluidic system was investigated under perfusion conditions with an adipogenic medium or an adipogenic medium supplemented with supernatant from differentiating ASCs (conditioned medium). Conditioned medium increased adipogenic differentiation compared to adipogenic medium with respect to accumulation of lipid-filled vacuoles and gene expression of key adipogenic markers (C/EBPα, C/EBPβ, C/EBPδ, PPARγ, LPL and adiponectin). The positive effects of conditioned medium were observed early in the differentiation process. Conclusions Using different cell densities and microfluidic perfusion cell cultures to suppress the effects of cell-released factors, we have demonstrated the significant role played by auto- or paracrine signaling in adipocyte differentiation. The cell-released factor(s) were shown to act in the recruitment phase of the differentiation process. PMID:23723991

  15. High-frequency and hot-platen curing of medium-density fiberboards

    Treesearch

    Robert R. Stevens; George E. Woodson

    1977-01-01

    The effects of two curing methods-high-frequencey heating and hot-platen heating-on the properties of a ureaformaldehyde-bonded medium-density fiberboard prepared with a southern-hardwoods furnish (50% southern read oak, 25% mockernut hickory, and 25% sweetgum) were studied. Boards of three densities-38, 44, and 50 lb./ft3-were cured by the two...

  16. Medium factors on anaerobic production of rhamnolipids by Pseudomonas aeruginosa SG and a simplifying medium for in situ microbial enhanced oil recovery applications.

    PubMed

    Zhao, Feng; Zhou, Jidong; Han, Siqin; Ma, Fang; Zhang, Ying; Zhang, Jie

    2016-04-01

    Aerobic production of rhamnolipid by Pseudomonas aeruginosa was extensively studied. But effect of medium composition on anaerobic production of rhamnolipid by P. aeruginosa was unknown. A simplifying medium facilitating anaerobic production of rhamnolipid is urgently needed for in situ microbial enhanced oil recovery (MEOR). Medium factors affecting anaerobic production of rhamnolipid were investigated using P. aeruginosa SG (Genbank accession number KJ995745). Medium composition for anaerobic production of rhamnolipid by P. aeruginosa is different from that for aerobic production of rhamnolipid. Both hydrophobic substrate and organic nitrogen inhibited rhamnolipid production under anaerobic conditions. Glycerol and nitrate were the best carbon and nitrogen source. The commonly used N limitation under aerobic conditions was not conducive to rhamnolipid production under anaerobic conditions because the initial cell growth demanded enough nitrate for anaerobic respiration. But rhamnolipid was also fast accumulated under nitrogen starvation conditions. Sufficient phosphate was needed for anaerobic production of rhamnolipid. SO4(2-) and Mg(2+) are required for anaerobic production of rhamnolipid. Results will contribute to isolation bacteria strains which can anaerobically produce rhamnolipid and medium optimization for anaerobic production of rhamnolipid. Based on medium optimization by response surface methodology and ions composition of reservoir formation water, a simplifying medium containing 70.3 g/l glycerol, 5.25 g/l NaNO3, 5.49 g/l KH2PO4, 6.9 g/l K2HPO4·3H2O and 0.40 g/l MgSO4 was designed. Using the simplifying medium, 630 mg/l of rhamnolipid was produced by SG, and the anaerobic culture emulsified crude oil to EI24 = 82.5 %. The simplifying medium was promising for in situ MEOR applications.

  17. Hyperconcentrated Sweet Whey, a New Culture Medium That Enhances Propionibacterium freudenreichii Stress Tolerance.

    PubMed

    Huang, Song; Rabah, Houem; Jardin, Julien; Briard-Bion, Valérie; Parayre, Sandrine; Maillard, Marie-Bernadette; Le Loir, Yves; Chen, Xiao Dong; Schuck, Pierre; Jeantet, Romain; Jan, Gwénaël

    2016-08-01

    Propionibacterium freudenreichii is used as a cheese-ripening starter and as a probiotic. Its reported physiological effects at the gut level, including modulation of bifidobacteria, colon epithelial cell proliferation and apoptosis, and intestinal inflammation, rely on active metabolism in situ Survival and activity are thus key factors determining its efficacy, creating stress adaptation and tolerance bottlenecks for probiotic applications. Growth media and growth conditions determine tolerance acquisition. We investigated the possibility of using sweet whey, a dairy by-product, to sustain P. freudenreichii growth. It was used at different concentrations (dry matter) as a culture medium. Using hyperconcentrated sweet whey led to enhanced multistress tolerance acquisition, overexpression of key stress proteins, and accumulation of intracellular storage molecules and compatible solutes, as well as enhanced survival upon spray drying. A simplified process from growth to spray drying of propionibacteria was developed using sweet whey as a 2-in-1 medium to both culture P. freudenreichii and protect it from heat and osmotic injury without harvesting and washing steps. As spray drying is far cheaper and more energy efficient than freeze-drying, this work opens new perspectives for the sustainable development of new starter and probiotic preparations with enhanced robustness. In this study, we demonstrate that sweet whey, a dairy industry by-product, not only allows the growth of probiotic dairy propionibacteria, but also triggers a multitolerance response through osmoadaptation and general stress response. We also show that propionibacteria accumulate compatible solutes under these culture conditions, which might account for the limited loss of viability after spray drying. This work opens new perspectives for more energy-efficient production of dairy starters and probiotics. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  18. Hyperconcentrated Sweet Whey, a New Culture Medium That Enhances Propionibacterium freudenreichii Stress Tolerance

    PubMed Central

    Huang, Song; Rabah, Houem; Jardin, Julien; Briard-Bion, Valérie; Parayre, Sandrine; Maillard, Marie-Bernadette; Le Loir, Yves; Schuck, Pierre; Jeantet, Romain

    2016-01-01

    ABSTRACT Propionibacterium freudenreichii is used as a cheese-ripening starter and as a probiotic. Its reported physiological effects at the gut level, including modulation of bifidobacteria, colon epithelial cell proliferation and apoptosis, and intestinal inflammation, rely on active metabolism in situ. Survival and activity are thus key factors determining its efficacy, creating stress adaptation and tolerance bottlenecks for probiotic applications. Growth media and growth conditions determine tolerance acquisition. We investigated the possibility of using sweet whey, a dairy by-product, to sustain P. freudenreichii growth. It was used at different concentrations (dry matter) as a culture medium. Using hyperconcentrated sweet whey led to enhanced multistress tolerance acquisition, overexpression of key stress proteins, and accumulation of intracellular storage molecules and compatible solutes, as well as enhanced survival upon spray drying. A simplified process from growth to spray drying of propionibacteria was developed using sweet whey as a 2-in-1 medium to both culture P. freudenreichii and protect it from heat and osmotic injury without harvesting and washing steps. As spray drying is far cheaper and more energy efficient than freeze-drying, this work opens new perspectives for the sustainable development of new starter and probiotic preparations with enhanced robustness. IMPORTANCE In this study, we demonstrate that sweet whey, a dairy industry by-product, not only allows the growth of probiotic dairy propionibacteria, but also triggers a multitolerance response through osmoadaptation and general stress response. We also show that propionibacteria accumulate compatible solutes under these culture conditions, which might account for the limited loss of viability after spray drying. This work opens new perspectives for more energy-efficient production of dairy starters and probiotics. PMID:27235433

  19. CNTF-ACM promotes mitochondrial respiration and oxidative stress in cortical neurons through upregulating L-type calcium channel activity.

    PubMed

    Sun, Meiqun; Liu, Hongli; Xu, Huanbai; Wang, Hongtao; Wang, Xiaojing

    2016-09-01

    A specialized culture medium termed ciliary neurotrophic factor-treated astrocyte-conditioned medium (CNTF-ACM) allows investigators to assess the peripheral effects of CNTF-induced activated astrocytes upon cultured neurons. CNTF-ACM has been shown to upregulate neuronal L-type calcium channel current activity, which has been previously linked to changes in mitochondrial respiration and oxidative stress. Therefore, the aim of this study was to evaluate CNTF-ACM's effects upon mitochondrial respiration and oxidative stress in rat cortical neurons. Cortical neurons, CNTF-ACM, and untreated control astrocyte-conditioned medium (UC-ACM) were prepared from neonatal Sprague-Dawley rat cortical tissue. Neurons were cultured in either CNTF-ACM or UC-ACM for a 48-h period. Changes in the following parameters before and after treatment with the L-type calcium channel blocker isradipine were assessed: (i) intracellular calcium levels, (ii) mitochondrial membrane potential (ΔΨm), (iii) oxygen consumption rate (OCR) and adenosine triphosphate (ATP) formation, (iv) intracellular nitric oxide (NO) levels, (v) mitochondrial reactive oxygen species (ROS) production, and (vi) susceptibility to the mitochondrial complex I toxin rotenone. CNTF-ACM neurons displayed the following significant changes relative to UC-ACM neurons: (i) increased intracellular calcium levels (p < 0.05), (ii) elevation in ΔΨm (p < 0.05), (iii) increased OCR and ATP formation (p < 0.05), (iv) increased intracellular NO levels (p < 0.05), (v) increased mitochondrial ROS production (p < 0.05), and (vi) increased susceptibility to rotenone (p < 0.05). Treatment with isradipine was able to partially rescue these negative effects of CNTF-ACM (p < 0.05). CNTF-ACM promotes mitochondrial respiration and oxidative stress in cortical neurons through elevating L-type calcium channel activity.

  20. 40 CFR 86.231-94 - Vehicle preparation.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 19 2012-07-01 2012-07-01 false Vehicle preparation. 86.231-94 Section... (CONTINUED) CONTROL OF EMISSIONS FROM NEW AND IN-USE HIGHWAY VEHICLES AND ENGINES Emission Regulations for 1994 and Later Model Year Gasoline-Fueled New Light-Duty Vehicles, New Light-Duty Trucks and New Medium...

  1. 40 CFR 86.231-94 - Vehicle preparation.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 19 2013-07-01 2013-07-01 false Vehicle preparation. 86.231-94 Section... (CONTINUED) CONTROL OF EMISSIONS FROM NEW AND IN-USE HIGHWAY VEHICLES AND ENGINES Emission Regulations for 1994 and Later Model Year Gasoline-Fueled New Light-Duty Vehicles, New Light-Duty Trucks and New Medium...

  2. 40 CFR 86.231-94 - Vehicle preparation.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 18 2011-07-01 2011-07-01 false Vehicle preparation. 86.231-94 Section... (CONTINUED) CONTROL OF EMISSIONS FROM NEW AND IN-USE HIGHWAY VEHICLES AND ENGINES Emission Regulations for 1994 and Later Model Year Gasoline-Fueled New Light-Duty Vehicles, New Light-Duty Trucks and New Medium...

  3. 40 CFR 86.231-94 - Vehicle preparation.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... (CONTINUED) CONTROL OF EMISSIONS FROM NEW AND IN-USE HIGHWAY VEHICLES AND ENGINES Emission Regulations for 1994 and Later Model Year Gasoline-Fueled New Light-Duty Vehicles, New Light-Duty Trucks and New Medium... 40 Protection of Environment 18 2010-07-01 2010-07-01 false Vehicle preparation. 86.231-94 Section...

  4. Polymers prepared from plant oils in liquid carbon dioxide

    USDA-ARS?s Scientific Manuscript database

    Recently, attention has been focused on the preparation of polymers in environmentally friendly media. Supercritical carbon dioxide (scCO2) is a promising candidate. The low toxicity of CO2 and lack of toxic solvent residues in the final products make CO2 an attractive medium for the synthesis of ...

  5. A New Approach to Prepare Vegetable Oil-Based Polymers

    USDA-ARS?s Scientific Manuscript database

    Polymers from vegetable oils, such as soybean oil, were prepared by cationic polymerization in supercritical carbon dioxide (scCO2) medium. Boron trifluoride diethyl etherate (BF3.OEt2) was selected as catalyst. The resulting polymers have molecular weight ranging from 21,842 to 118,300 g/mol. Nu...

  6. Underground Coal Preparation System and Applications

    NASA Astrophysics Data System (ADS)

    Wei, Cao; DeYong, Shang; BaoNing, Zhang

    2018-03-01

    The underground coal preparation is a cutting-edge technology of the coal industry worldwide. This paper introduced the meaning of implementing the underground coal preparation, and the practical applications of underground mechanical moving screen jig, underground heavy medium shallow slot and underground air jigger. Through analyzing the main separation equipment and the advantages and disadvantages of three primary processes from aspects of process complexity, slime water treatment, raw coal preparation, etc., the difference among technology investment, construction scale, production cost and economic benefit is concluded.

  7. Induction of endothelial cell proliferation by angiogenic factors released by activated monocytes

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

    Pakala, Rajbabu; Watanabe, Takuya; Benedict, Claude R

    2002-06-01

    Introduction: Cell-cell interaction is an essential component of atherosclerotic plaque development. Activated monocytes appear to play a central role in the development of atherosclerosis, not only through foam cell formation but also via the production of various growth factors that induce proliferation of different cell types that are involved in the plaque development. Using serum free co-culture method, we determined the effect of monocytes on endothelial cell proliferation. Methods: Endothelial cell proliferation is determined by the amount of [{sup 3}H]thymidine incorporated in to the DNA. Basic fibroblast growth factor (b-FGF), vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) levels inmore » the conditioned medium were determined by ELISA. Results: Conditioned medium from unactivated monocytes partially inhibited endothelial cell proliferation, whereas conditioned medium from activated monocytes promoted endothelial cell proliferation. The mitogenic effect of conditioned medium derived from activated monocytes is due to the presence of b-FGF, VEGF and IL-8. Neutralizing antibodies against b-FGF, VEGF and IL-8 partially reversed the mitogenic effect of conditioned medium derived from activated monocytes. When b-FGF, VEGF and IL-8 were immunoprecipitated from conditioned medium derived from activated monocytes, it is less mitogenic to endothelial cells. Conclusion: Activated monocytes may play an important role in the development of atherosclerotic plaque by producing endothelial cell growth factors.« less

  8. Mechanical fibrinogen-depletion supports heparin-free mesenchymal stem cell propagation in human platelet lysate.

    PubMed

    Laner-Plamberger, Sandra; Lener, Thomas; Schmid, Doris; Streif, Doris A; Salzer, Tina; Öller, Michaela; Hauser-Kronberger, Cornelia; Fischer, Thorsten; Jacobs, Volker R; Schallmoser, Katharina; Gimona, Mario; Rohde, Eva

    2015-11-10

    Pooled human platelet lysate (pHPL) is an efficient alternative to xenogenic supplements for ex vivo expansion of mesenchymal stem cells (MSCs) in clinical studies. Currently, porcine heparin is used in pHPL-supplemented medium to prevent clotting due to plasmatic coagulation factors. We therefore searched for an efficient and reproducible medium preparation method that avoids clot formation while omitting animal-derived heparin. We established a protocol to deplete fibrinogen by clotting of pHPL in medium, subsequent mechanical hydrogel disruption and removal of the fibrin pellet. After primary culture, bone-marrow and umbilical cord derived MSCs were tested for surface markers by flow cytometry and for trilineage differentiation capacity. Proliferation and clonogenicity were analyzed for three passages. The proposed clotting procedure reduced fibrinogen more than 1000-fold, while a volume recovery of 99.5 % was obtained. All MSC types were propagated in standard and fibrinogen-depleted medium. Flow cytometric phenotype profiles and adipogenic, osteogenic and chondrogenic differentiation potential in vitro were independent of MSC-source or medium type. Enhanced proliferation of MSCs was observed in the absence of fibrinogen but presence of heparin compared to standard medium. Interestingly, this proliferative response to heparin was not detected after an initial contact with fibrinogen during the isolation procedure. Here, we present an efficient, reproducible and economical method in compliance to good manufacturing practice for the preparation of MSC media avoiding xenogenic components and suitable for clinical studies.

  9. [Massive multiplication of coffee (Coffee arabica L. cv. Catimor) through embryogenic cell suspension culture].

    PubMed

    Flermoso-Gallardo, L; Menóndez-Yuffá, A

    2000-01-01

    Cell suspensions offer several advantages as a system for massive propagation because of the high rates of multiplication, the higher homogeneity in the culture conditions and the possibility of automatization. In this study, different experimental conditions were analyzed to establish embryogenic cell suspension cultures of coffee. The best conditions to establish the embryogenic cell suspension cultures of coffee were as follows: coffee leaf sections were cultivated during 12 weeks (Stage I) in a solid medium with the Murashige and Skoog salts, 2 mg/l kinetin and 0.5 mg/l 2,4-dichlorophenoxiacetic acid (medium 1). Under these conditions the explants formed a callus tissue that was transferred to a liquid medium containing 5 mg/l of 6-benzylamlno-purine (medium 2). After 12 days in a shaking liquid medium (Stage II), the cultures were sieved and were maintained In the same media, which was renewed every eight days (Stage III). This method yielded 1884 embryos in 50 ml; placing the embryos under conditions for germination yielded plantlets of normal appearance.

  10. Optimization of Carbon and Nitrogen Sources for Extracellular Polymeric Substances Production by Chryseobacterium indologenes MUT.2.

    PubMed

    Khani, Mojtaba; Bahrami, Ali; Chegeni, Asma; Ghafari, Mohammad Davoud; Mansouran Zadeh, ALi

    2016-06-01

    Bacterial Extracellular Polymeric Substances (EPS) are environmental friendly and versatile polymeric materials that are used in a wide range of industries such as: food, textile, cosmetics, and pharmaceuticals. To make the production process of the EPS cost-effective, improvements in the production yield is required which could be implemented through application of processes such as optimized culture conditions, and development of the strains with higher yield ( e.g . through genetic manipulation), or using low-cost substrates. In this work, the effects of carbon and nitrogen sources were studied in order to improve the EPS production by the submerged cultivation of Chryseobacterium indologenes MUT.2. The mesophilic microorganism Chryseobacterium indologenes MUT.2, was grown and maintained in the Luria Bertani agar. The initial basal medium contained: glucose (20 g.L -1 ), yeast extracts (5 g.L -1 ), K 2 HPO 4 (6 g.L -1 ), NaH 2 PO 4 (7 g.L -1 ), NH 4 CL (0.7 g.L -1 ), and MgSO 4 (0.5 g.L -1 ). For evaluating the carbon and nitrogen sources' effect on the fermentation performance, cultures were prepared in 500 mL flasks filled with 300 mL of the medium. The single-factor experiments based on statistics was employed to evaluate and optimize the carbon and nitrogen sources for EPS production in the liquid culture medium of Chryseobacterium indologenes MUT.2. The preferred carbon-sources, sucrose and glucose, commonly gave the highest EPS production of 8.32 and 6.37 g.L -1 , respectively, and the maximum EPS production of 8.87 g.L -1 was achieved when glutamic acid (5 g.L -1 ) was employed as the nitrogen source. In this work, the culture medium for production of EPS by Chryseobacterium indologenes MUT.2 was optimized. Compared to the basal culture medium in shake-flasks and stirred tank bioreactor, the use of optimized culture medium has resulted in a 53% and 73% increase in the EPS production, respectively.

  11. Selenium Nanoparticles Formed by Modulation of Carrageenan Enhance Osteogenic Differentiation of Mesenchymal Stem Cells.

    PubMed

    Kim, Jin; Lee, Ki-Young; Lee, Chang-Moon

    2016-03-01

    Fabrication of nano-sized selenium (Se) particles may help to expend the applications of Se. In this study, we focused on the preparation and characterization of Se nanoparticles (Se NPs) modulated with carrageenan (CA). Furthermore, their influence on osteoblast cell growth was investigated in vitro. Spherical Se-NPs, of 100-200 nm diameter, were prepared simply by adding κ-, ι-, and λ-CA, which has sulfate groups, hydroxyl groups, and carboxyl groups. CA-modulated Se NPs (CA-Se NPs) were readily suspended in liquid medium with no precipitation over long time periods. In particular, it was found through Alizarin Red S staining that the growth of osteoblast D1 cells treated with λ-CA-Se NPs was improved significantly. These results suggest that Se NPs can be prepared simply, using CA, have good suspension stability in liquid medium, and λ-CA-Se NPs may induce the growth of osteoblast cells.

  12. Epithelial cell morphology and adhesion on diamond films deposited and chemically modified by plasma processes.

    PubMed

    Rezek, Bohuslav; Ukraintsev, Egor; Krátká, Marie; Taylor, Andrew; Fendrych, Frantisek; Mandys, Vaclav

    2014-09-01

    The authors show that nanocrystalline diamond (NCD) thin films prepared by microwave plasma enhanced chemical vapor deposition apparatus with a linear antenna delivery system are well compatible with epithelial cells (5637 human bladder carcinoma) and significantly improve the cell adhesion compared to reference glass substrates. This is attributed to better adhesion of adsorbed layers to diamond as observed by atomic force microscopy (AFM) beneath the cells. Moreover, the cell morphology can be adjusted by appropriate surface treatment of diamond by using hydrogen and oxygen plasma. Cell bodies, cytoplasmic rims, and filopodia were characterized by Peakforce AFM. Oxidized NCD films perform better than other substrates under all conditions (96% of cells adhered well). A thin adsorbed layer formed from culture medium and supplemented with fetal bovine serum (FBS) covered the diamond surface and played an important role in the cell adhesion. Nevertheless, 50-100 nm large aggregates formed from the RPMI medium without FBS facilitated cell adhesion also on hydrophobic hydrogenated NCD (increase from 23% to 61%). The authors discuss applicability for biomedical uses.

  13. Preparation of Ion Exchange Films for Solid-Phase Spectrophotometry and Solid-Phase Fluorometry

    NASA Technical Reports Server (NTRS)

    Hill, Carol M.; Street, Kenneth W.; Tanner, Stephen P.; Philipp, Warren H.

    2000-01-01

    Atomic spectroscopy has dominated the field of trace inorganic analysis because of its high sensitivity and selectivity. The advantages gained by the atomic spectroscopies come with the disadvantage of expensive and often complicated instrumentation. Solid-phase spectroscopy, in which the analyte is preconcentrated on a solid medium followed by conventional spectrophotometry or fluorometry, requires less expensive instrumentation and has considerable sensitivity and selectivity. The sensitivity gains come from preconcentration and the use of chromophore (or fluorophore) developers and the selectivity is achieved by use of ion exchange conditions that favor the analyte in combination with speciative chromophores. Little work has been done to optimize the ion exchange medium (IEM) associated with these techniques. In this report we present a method for making ion exchange polymer films, which considerably simplify the solid-phase spectroscopic techniques. The polymer consists of formaldehyde-crosslinked polyvinyl alcohol with polyacrylic acid entrapped therein. The films are a carboxylate weak cation exchanger in the calcium form. They are mechanically sturdy and optically transparent in the ultraviolet and visible portion of the spectrum, which makes them suitable for spectrophotometry and fluorometry.

  14. Xylanase production with xylan rich lignocellulosic wastes by a local soil isolate of Trichoderma viride

    PubMed Central

    Goyal, Meenakshi; Kalra, K.L.; Sareen, V.K.; Soni, G.

    2008-01-01

    In the present study, cultural and nutritional conditions for enhanced production of xylanase by a local soil isolate of Trichoderma viride, using various lignocellulosic substrates in submerged culture fermentation have been optimized. Of the lignocellulosics used, maize straw was the best inducer followed by jowar straw for xylanase production. The highest activity achieved was between 14 to 17 days of fermentation. A continuous increase in xylanase production was observed with increasing level of lignocellulosics in the medium and highest activity was observed with maize straw at 5% level. Xylanase production with higher levels of lignocellulosics (3 to 5%) of maize, jowar and barseem was found to be higher as compared to that with commercial xylan as carbon source. Sodium nitrate was the best nitrogen source among the six sources used. Maximum xylanase production was achieved with initial medium pH of 3.5–4.0 and incubation temperature of 25ºC.The enzyme preparation was effective in bringing about saccharification of different lignocellulosics. The xylanase production could be further improved by using alkali treated straw as carbon source. PMID:24031262

  15. Dynamics of entropy and nonclassical properties of the state of a Λ-type three-level atom interacting with a single-mode cavity field with intensity-dependent coupling in a Kerr medium

    NASA Astrophysics Data System (ADS)

    Faghihi, M. J.; Tavassoly, M. K.

    2012-02-01

    In this paper, we study the interaction between a three-level atom and a quantized single-mode field with ‘intensity-dependent coupling’ in a ‘Kerr medium’. The three-level atom is considered to be in a Λ-type configuration. Under particular initial conditions, which may be prepared for the atom and the field, the dynamical state vector of the entire system will be explicitly obtained, for the arbitrary nonlinearity function f(n) associated with any physical system. Then, after evaluating the variation of the field entropy against time, we will investigate the quantum statistics as well as some of the nonclassical properties of the introduced state. During our calculations we investigate the effects of intensity-dependent coupling, Kerr medium and detuning parameters on the depth and domain of the nonclassicality features of the atom-field state vector. Finally, we compare our obtained results with those of V-type three-level atoms.

  16. Methods for the cultivation of ciliated protozoa from the large intestine of horses.

    PubMed

    Bełżecki, Grzegorz; Miltko, Renata; Michałowski, Tadeusz; McEwan, Neil R

    2016-01-01

    This paper describes cultivation methods for ciliates from the digestive tract of horses. Members of three different genera were successfully grown in vitro for short periods of time. However, only cells belonging to the genus Blepharocorys, which resides in the horse's large intestine, were maintained for longer periods. This Blepharocorys culture was successfully grown in vitro after inoculation of freshly excreted horse faeces in culture medium containing a population of bacteria. The ciliates survived for over six months, and the density of their population varied between 1.7 × 10(3) and 2.4 × 10(3) cells mL(-1). Favourable conditions for the prolonged cultivation of this ciliate were observed when the medium was prepared by mixing horse faeces and 'caudatum' salt solution in a 1:1 V/V ratio together with food (60% powdered meadow hay, 16% wheat gluten, 12% barley flour and 12% microcrystalline cellulose) supplied as 0.20 mg mL(-1) culture per day. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  17. Medium voltage therapy for preventing and treating asystole and PEA in ICDs.

    PubMed

    Gilman, Byron L; Brewer, James E; Kroll, Kai; Kroll, Mark W

    2009-01-01

    Sudden cardiac death (SCD) takes up to 500,000 lives each year before a victim can even be treated. To address this the implantable cardioverter defibrillator (ICD) was developed to treat those identified at high risk of SCD. Unfortunately, there are a significant number of cases in which the ICD does not successfully return a victim to normal rhythm and effective perfusion of the blood. The vast majority of cases that are not responsive to the ICD therapy require cardio-pulmonary resuscitation (CPR) according to current resuscitation guidelines. A novel electrical stimulus called medium voltage therapy (MVT) has shown efficacy in producing coronary and carotid blood flow during ventricular fibrillation. This report presents the case that the same stimulus may be effective and feasible for use in ICD patients that do not respond to their ICD therapy, or do not have a rhythm in which, an ICD shock is indicated. The inclusion of MVT technology in implantable devices may be effective in preparing the heart for successful defibrillation or in improving the metabolic condition of the heart to the extent that a pulsatile rhythm may spontaneously develop.

  18. Hydrogen photoproduction in green algae Chlamydomonas reinhardtii sustainable over 2 weeks with the original cell culture without supply of fresh cells nor exchange of the whole culture medium.

    PubMed

    Yagi, Takafumi; Yamashita, Kyohei; Okada, Norihide; Isono, Takumi; Momose, Daisuke; Mineki, Shigeru; Tokunaga, Eiji

    2016-07-01

    Unicellular green algae Chlamydomonas reinhardtii are known to make hydrogen photoproduction under the anaerobic condition with water molecules as the hydrogen source. Since the hydrogen photoproduction occurs for a cell to circumvent crisis of its survival, it is only temporary. It is a challenge to realize persistent hydrogen production because the cells must withstand stressful conditions to survive with alternation of generations in the cell culture. In this paper, we have found a simple and cost-effective method to sustain the hydrogen production over 14 days in the original culture, without supply of fresh cells nor exchange of the culture medium. This is achieved for the cells under hydrogen production in a sulfur-deprived culture solution on the {anaerobic, intense light} condition in a desiccator, by periodically providing a short period of the recovery time (2 h) with a small amount of TAP(+S) supplied outside of the desiccator. As this operation is repeated, the response time of transition into hydrogen production (preparation time) is shortened and the rate of hydrogen production (build up time) is increased. The optimum states of these properties favorable to the hydrogen production are attained in a few days and stably sustained for more than 10 days. Since generations are alternated during this consecutive hydrogen production experiment, it is suggested that the improved hydrogen production properties are inherited to next generations without genetic mutation. The properties are reset only when the cells are placed on the {sulfur-sufficient, aerobic, moderate light} conditions for a long time (more than 1 day at least).

  19. Comparative Study of the Effect of Defects on Selective Adsorption of Butanol from Butanol/Water Binary Vapor Mixtures in Silicalite-1 Films

    DOE PAGES

    Farzaneh, Amirfarrokh; DeJaco, Robert F.; Ohlin, Lindsay; ...

    2017-08-02

    A promising route for sustainable 1-butanol (butanol) production is ABE (acetone, butanol, ethanol) fermentation. However, recovery of the products is challenging because of the low concentrations obtained in the aqueous solution, thus hampering large-scale production of biobutanol. Membrane and adsorbent-based technologies using hydrophobic zeolites are interesting alternatives to traditional separation techniques (e.g., distillation) for energy-efficient separation of butanol from aqueous mixtures. To maximize the butanol over water selectivity of the material, it is important to reduce the number of hydrophilic adsorption sites. This can, for instance, be achieved by reducing the density of lattice defect sites where polar silanol groupsmore » are found. The density of silanol defects can be reduced by preparing the zeolite at neutral pH instead of using traditional synthesis solutions with high pH. In this work, binary adsorption of butanol and water in two silicalite-1 films was studied using in situ attenuated total reflectance–Fourier transform infrared (ATR-FTIR) spectroscopy under equal experimental conditions. One of the films was prepared in fluoride medium, whereas the other one was prepared at high pH using traditional synthesis conditions. The amounts of water and butanol adsorbed from binary vapor mixtures of varying composition were determined at 35 and 50 °C, and the corresponding adsorption selectivities were also obtained. Both samples showed very high selectivities (100–23 000) toward butanol under the conditions studied. The sample having low density of defects, in general, showed ca. a factor 10 times higher butanol selectivity than the sample having a higher density of defects at the same experimental conditions. This difference was due to a much lower adsorption of water in the sample with low density of internal defects. Analysis of molecular simulation trajectories provides insights on the local selectivities in the zeolite channel network and at the film surface.« less

  20. Comparative Study of the Effect of Defects on Selective Adsorption of Butanol from Butanol/Water Binary Vapor Mixtures in Silicalite-1 Films

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

    Farzaneh, Amirfarrokh; DeJaco, Robert F.; Ohlin, Lindsay

    A promising route for sustainable 1-butanol (butanol) production is ABE (acetone, butanol, ethanol) fermentation. However, recovery of the products is challenging because of the low concentrations obtained in the aqueous solution, thus hampering large-scale production of biobutanol. Membrane and adsorbent-based technologies using hydrophobic zeolites are interesting alternatives to traditional separation techniques (e.g., distillation) for energy-efficient separation of butanol from aqueous mixtures. To maximize the butanol over water selectivity of the material, it is important to reduce the number of hydrophilic adsorption sites. This can, for instance, be achieved by reducing the density of lattice defect sites where polar silanol groupsmore » are found. The density of silanol defects can be reduced by preparing the zeolite at neutral pH instead of using traditional synthesis solutions with high pH. In this work, binary adsorption of butanol and water in two silicalite-1 films was studied using in situ attenuated total reflectance–Fourier transform infrared (ATR-FTIR) spectroscopy under equal experimental conditions. One of the films was prepared in fluoride medium, whereas the other one was prepared at high pH using traditional synthesis conditions. The amounts of water and butanol adsorbed from binary vapor mixtures of varying composition were determined at 35 and 50 °C, and the corresponding adsorption selectivities were also obtained. Both samples showed very high selectivities (100–23 000) toward butanol under the conditions studied. The sample having low density of defects, in general, showed ca. a factor 10 times higher butanol selectivity than the sample having a higher density of defects at the same experimental conditions. This difference was due to a much lower adsorption of water in the sample with low density of internal defects. Analysis of molecular simulation trajectories provides insights on the local selectivities in the zeolite channel network and at the film surface.« less

  1. A xyloglucan from seeds of the native Brazilian species Hymenaea courbaril for micropropagation of Marubakaido and Jonagored apples.

    PubMed

    Lima-Nishimura, N; Quoirin, M; Naddaf, Y G; Wilhelm, H M; Ribas, L L F; Sierakowski, M-R

    2003-01-01

    Xyloglucan was extracted from seeds of Hymenaea courbaril and mixed with agar to prepare a solid culture medium used for micropropagation of the Marubakaido apple rootstock (Malus prunifolia Borkh) and cv. Jonagored (Malus domestica). The performance on gels created from a blend of 0.4%agar and 0.2% xyloglucan (w/v) was compared with that on media gelled with a standard concentration 0.6% (w/v) of agar. The growth of shoots and the multiplication rate were higher on the modified culture medium than on the agar-gelled medium. The occurrence of hyperhydric shoots was lower on the modified medium. In the absence of auxin, shoot rooting reached 70% (Marubakaido) and 66% (Jonagored) on the agar-xyloglucan medium and 6.7% and 10.4%, respectively, on the agar medium. When 0.25 microM indole-3-butyric acid (IBA) was added to both media, the modified medium gave better results in terms of rooting percentage and quality of roots than the agar-gelled medium.

  2. Porous structures of polymer films prepared by spin coating with mixed solvents under humid condition.

    PubMed

    Park, Min Soo; Joo, Wonchul; Kim, Jin Kon

    2006-05-09

    We investigate the effects of interfacial energy between water and solvent as well as polymer concentration on the formation of porous structures of polymer films prepared by spin coating of cellulose acetate butyrate (CAB) in mixed solvent of tetrahydrofuran (THF) and chloroform under humid condition. The interfacial energy between water and the solvent was gradually changed by the addition of chloroform to the solvent. At a high polymer concentration (0.15 g/cm3 in THF), porous structures were limited only at the top surfaces of CAB films, regardless of interfacial energies, due to the high viscosity of the solution. At a medium concentration (approximately 0.08 g/cm3 in THF), CAB film had relatively uniform pores at the top surface and very small pores inside the film because of the mixing of the water droplets with THF solution. When chloroform was added to THF, pores at the inner CAB film had a comparable size with those at the top surface because of the reduced degree of the mixing between the water droplets and the mixed solvent. A further decrease in polymer concentration (0.05 g/cm3 in THF) caused the final films to have a two-layer porous structure, and the size of pores at each layer was almost the same.

  3. An investigation into the stability of commercial versus MG63-derived hepatocyte growth factor under flow cultivation conditions.

    PubMed

    Meneghello, Giulia; Storm, Michael P; Chaudhuri, Julian B; De Bank, Paul A; Ellis, Marianne J

    2015-03-01

    The scale-up of tissue engineering cell culture must ensure that conditions are maintained while also being cost effective. Here we analyse the stability of hepatocyte growth factor (HGF) to investigate whether concentrations change under dynamic conditions, and compare commercial recombinant human HGF as an additive in 'standard medium', to HGF secreted by the osteosarcoma cell line MG63 as a 'preconditioned medium'. After 3 h under flow conditions, HGF in the standard medium degraded to 40% of its original concentration but HGF in the preconditioned medium remained at 100%. The concentration of secreted HGF was 10 times greater than the working concentration of commercially-available HGF. Thus HGF within this medium has increased stability; MG63-derived HGF should therefore be investigated as a cost-effective alternative to current lyophilised powders for use in in vitro models. Furthermore, we recommend that those intending to use HGF (or other growth factors) should consider similar stability testing before embarking on experiments with media flow.

  4. Monomer Derived Poly(Furfuryl)/BaTiO3 0-3 Nanocomposite Capacitors: Maximization of the Effective Permittivity Through Control at the Interface.

    PubMed

    Pearsall, Frederick A; Lombardi, Julien; O'Brien, Stephen

    2017-11-22

    Frequency stable, high permittivity nanocomposite capacitors produced under mild processing conditions offer an attractive replacement to MLCCs derived from conventional ceramic firing. Here, 0-3 nanocomposites were prepared using gel-collection derived barium titanate nanocrystals, suspended in a poly(furfuryl alcohol) matrix, resulting in a stable, high effective permittivity, low loss dielectric. The nanocrystals are produced at 60 °C, emerging as fully crystallized cubic BTO, 8 nm, paraelectric with a highly functional surface that enables both suspension and chemical reaction in organic solvents. The nanocrystals were suspended in furfuryl alcohol inside a uniquely prepared mold, in which volume fraction of nanocrystal filler (ν f ) could be varied. Polymerization of the matrix in situ at 70-90 °C resulted in a nanocomposite with a higher than anticipated effective permittivity (up to 50, with ν f only 0.41, 0.5-2000 kHz), exceptional stability as a function of frequency, and very favorable dissipation factors (tan δ < 0.01, ν f < 0.41; tan δ < 0.05, ν f < 0.5). The increased permittivity is attributed to the covalent attachment of the poly(furfuryl alcohol) matrix to the surface of the nanocrystals, homogenizing the particle-matrix interface, limiting undercoordinated surface sites and reducing void space. XPS and FTIR confirmed strong interfacial interaction between matrix and nanocrystal surface. Effective medium approximations were used to compare this with similar nanocomposite systems. It was found that the high effective permittivity could not be attributed to the combination of two components alone, rather the creation of a hybrid nanocomposite possessing its own dielectric behavior. A nondispersive medium was selected to focus on the frequency dependent permittivity of the 8 nm barium titanate nanocrystals. Experimental corroboration with known theory is evident until a specific volume fraction (ν f ≈ 0.3) where, due to a sharp increase in the effective permittivity, approximations fail to adequately describe the nanocomposite medium.

  5. Evidence for the lack of spare high-affinity insulin receptors in skeletal muscle.

    PubMed Central

    Camps, M; Gumà, A; Viñals, F; Testar, X; Palacín, M; Zorzano, A

    1992-01-01

    In this study, the relationship between the concentration of extracellular insulin, insulin binding and insulin action was evaluated in skeletal muscle. Initially we investigated the dose-response relationship of insulin action using three different experimental models that are responsive to insulin, i.e. the isolated perfused rat hindquarter, incubated strips of soleus muscle, and insulin receptors partially affinity-purified from skeletal muscle. We selected as insulin-sensitive parameters glucose uptake in the perfused hindquarter, lactate production in the incubated muscle preparation, and tyrosine receptor kinase activity in the purified receptor preparation. Our results showed that the dose-response curves obtained in the perfused hindquarter and in the incubated muscle were superimposable. In contrast, the dose-response curve for insulin-stimulated receptor tyrosine kinase activity in partially purified receptors was displaced to the left compared with the curves obtained in the perfused hindquarter and in the incubated muscle. The differences between the dose-response curve for receptor tyrosine kinase and those for glucose uptake and lactate production were not explained by a substantial insulin concentration gradient between medium and interstitial space. Thus the medium/interstitial insulin concentration ratio, when assayed in the incubated intact muscle at 5 degrees C, was close to 1. We also compared the dose-response curve of insulin-stimulated receptor tyrosine kinase with the pattern of insulin-binding-site occupancy. The curve of insulin-stimulated receptor kinase activity fitted closely with the occupancy of high-affinity binding sites. In summary, assuming that the estimation of the medium/interstitial insulin concentration ratio obtained at 5 degrees C reflects the actual ratio under more physiological conditions, our results suggest that maximal insulin action is obtained in skeletal muscle at insulin concentrations which do allow full occupancy of high-affinity binding sites. Therefore our data provide evidence for a lack of spare high-affinity insulin receptors in skeletal muscle. PMID:1323279

  6. Evidence for the lack of spare high-affinity insulin receptors in skeletal muscle.

    PubMed

    Camps, M; Gumà, A; Viñals, F; Testar, X; Palacín, M; Zorzano, A

    1992-08-01

    In this study, the relationship between the concentration of extracellular insulin, insulin binding and insulin action was evaluated in skeletal muscle. Initially we investigated the dose-response relationship of insulin action using three different experimental models that are responsive to insulin, i.e. the isolated perfused rat hindquarter, incubated strips of soleus muscle, and insulin receptors partially affinity-purified from skeletal muscle. We selected as insulin-sensitive parameters glucose uptake in the perfused hindquarter, lactate production in the incubated muscle preparation, and tyrosine receptor kinase activity in the purified receptor preparation. Our results showed that the dose-response curves obtained in the perfused hindquarter and in the incubated muscle were superimposable. In contrast, the dose-response curve for insulin-stimulated receptor tyrosine kinase activity in partially purified receptors was displaced to the left compared with the curves obtained in the perfused hindquarter and in the incubated muscle. The differences between the dose-response curve for receptor tyrosine kinase and those for glucose uptake and lactate production were not explained by a substantial insulin concentration gradient between medium and interstitial space. Thus the medium/interstitial insulin concentration ratio, when assayed in the incubated intact muscle at 5 degrees C, was close to 1. We also compared the dose-response curve of insulin-stimulated receptor tyrosine kinase with the pattern of insulin-binding-site occupancy. The curve of insulin-stimulated receptor kinase activity fitted closely with the occupancy of high-affinity binding sites. In summary, assuming that the estimation of the medium/interstitial insulin concentration ratio obtained at 5 degrees C reflects the actual ratio under more physiological conditions, our results suggest that maximal insulin action is obtained in skeletal muscle at insulin concentrations which do allow full occupancy of high-affinity binding sites. Therefore our data provide evidence for a lack of spare high-affinity insulin receptors in skeletal muscle.

  7. Effects of consumer food preparation on acrylamide formation.

    PubMed

    Jackson, Lauren S; Al-Taher, Fadwa

    2005-01-01

    Acrylamide is formed in high-carbohydrate foods during high temperature processes such as frying, baking, roasting and extrusion. Although acrylamide is known to form during industrial processing of food, high levels of the chemical have been found in home-cooked foods, mainly potato- and grain-based products. This chapter will focus on the effects of cooking conditions (e.g. time/temperature) on acrylamide formation in consumer-prepared foods, the use of surface color (browning) as an indicator of acrylamide levels in some foods, and methods for reducing acrylamide levels in home-prepared foods. As with commercially processed foods, acrylamide levels in home-prepared foods tend to increase with cooking time and temperature. In experiments conducted at the NCFST, we found that acrylamide levels in cooked food depended greatly on the cooking conditions and the degree of "doneness", as measured by the level of surface browning. For example, French fries fried at 150-190 degrees C for up to 10 min had acrylamide levels of 55 to 2130 microg/kg (wet weight), with the highest levels in the most processed (highest frying times/temperatures) and the most highly browned fries. Similarly, more acrylamide was formed in "dark" toasted bread slices (43.7-610.7 microg/kg wet weight), than "light" (8.27-217.5 microg/kg) or "medium" (10.9-213.7 microg/kg) toasted slices. Analysis of the surface color by colorimetry indicated that some components of surface color ("a" and "L" values) correlated highly with acrylamide levels. This indicates that the degree of surface browning could be used as an indicator of acrylamide formation during cooking. Soaking raw potato slices in water before frying was effective at reducing acrylamide levels in French fries. Additional studies are needed to develop practical methods for reducing acrylamide formation in home-prepared foods without changing the acceptability of these foods.

  8. Cultivation of Tomato Tissues Capable of Forming Flowers and Fruits in Vitro

    NASA Technical Reports Server (NTRS)

    Galston, Arthur W.

    1998-01-01

    The final phase of this research project was designed to develop a practical method for producing a steady supply of fresh cherry tomato fruits over a period of several months, for possible use as a fresh vegetable supplement to a standard diet of astronauts on extended missions. This effort was successful. We were able to excise immature flowers from Pixie tomato plants grown in a controlled condition room, implant them on artificial media under aseptic conditions, and get them to develop into edible fruits in a little over a month. The medium (Murashige-Skoog) was purchased from Sigma, supplemented with sugar plus a synthetic analog of the plant hormone cytokinin, and adjusted to pH 5.8. A temperature of at least 25 C and visible light helped to produce ripe red fruits within 7 weeks. To ensure a steady supply of such tomatoes, we found it possible to store the explanted flower buds in MS medium at 5 C for at least 6 weeks without significant loss of ability to develop into fruits. This means that many containers could be prepared before launch and put into a refrigerator; a convenient number could then be removed periodically to guarantee a succession of harvests during the life of an extended mission. Details are found in the attached reprints. Subsequent applications for funds for flight or continued research were denied, and the project was terminated.

  9. Silica hydride intermediate for octadecylsilica and phenyl bonded phase preparation via heterogeneous hydrosilation in supercritical carbon dioxide.

    PubMed

    Scully, N M; Ashu-Arrah, B A; Nagle, A P; Omamogho, J O; O'Sullivan, G P; Friebolin, V; Dietrich, B; Albert, K; Glennon, J D

    2011-04-15

    Investigations into the preparation of silica hydride intermediate in supercritical carbon dioxide (sc-CO(2)) that avoids the use of organic solvents such as toluene or dioxane are described. The effects of reaction temperature, pressure and time on the surface coverage of the supercritical fluid generated silica hydride intermediate were studied. Under optimised supercritical conditions of 120°C, 483 bar and 3 h reaction time, silica hydride (Si-H) conversion efficiencies of ca. 40% were achieved for the hydride intermediate prepared from a monofunctional silane reagent (dimethylmethoxysilane). Si-H conversion efficiencies (as determined from (29)Si CP-MAS NMR spectral analysis) for the hydride intermediate prepared from triethoxysilane (TES) in sc-CO(2) were found to be comparable to those obtained using a TES silanisation approach in an organic solvent. (13)C and (29)Si CP-MAS-NMR spectroscopy was employed to provide a complete structural assignment of the silica hydride intermediates. Furthermore, supercritical CO(2) was subsequently employed as a reaction medium for the heterogenous hydrosilation of silica hydride with octadecene and with styrene, in the presence of a free radical initiator. These supercritical fluid generated reversed-phase materials were prepared in a substantially reduced reaction time (3 h) compared to organic solvent based methods (100 h reaction time). Silica functionalisation in sc-CO(2) presents an efficient and clean alternative to organic solvent based methods for the preparation of important silica hydride intermediate and silica bonded stationary phases via a hydrosilation approach. Copyright © 2010 Elsevier B.V. All rights reserved.

  10. Transformation of proanthocyanidin A2 to its isomers under different physiological pH conditions and common cell culture medium.

    PubMed

    Lu, Wen-Chien; Huang, Wei-Ting; Kumaran, Alaganandam; Ho, Chi-Tang; Hwang, Lucy Sun

    2011-06-08

    Proanthocyanidins constitute an important class of polyphenols ubiquitously found in plants. They have been extensively studied for their antioxidant capacity and bioactivity in vitro and in animal models. However, their stability under different pH conditions and in cell culture medium has not been well documented. In the present study, it was observed that proanthocyanidin A2 (PA2) was relatively more stable in acidic condition than in weak alkaline condition. PA2 was also quite unstable in basal-Dulbecco's Modified Eagle medium (b-DMEM medium) at 37 °C. The addition of PA2 to the cell culture medium accelerated its epimerization with a half-life of <15 min, and ethylenediaminetetraacetic acid (EDTA) could not stop the reaction. The results also demonstrated that the major isomers transformed in the weak alkaline condition or cell culture medium at 37 °C were identified as epicatechin-(4β→8; 2β→O→7)-ent-catechin (proanthocyanidin A4) and epicatechin-(4β→6; 2β→O→7)-ent-catechin. The rates of transformation were dependent on the pH or the components of the medium. Therefore, the results obtained for PA2 in the cell culture bioassays, which were usually carried out for 24 h, might not represent the true activity of the original PA2. The stability and transformation of PA2 should be considered when the bioactivity of PA2 is evaluated in a given cell culture system.

  11. From oligomers to molecular giants of soybean oil in supercritical carbon dioxide medium: 1. Preparation of polymers with lower molecular weight from soybean oil.

    PubMed

    Liu, Zengshe; Sharma, Brajendra K; Erhan, Sevim Z

    2007-01-01

    Polymers with a low molecular weight derived from soybean oil have been prepared in a supercritical carbon dioxide medium by cationic polymerization. Boron trifluoride diethyl etherate was used as an initiator. Influences of polymerization temperature, amount of initiator, and carbon dioxide pressure on the molecular weight were investigated. It is shown that the higher polymerization temperature favors polymers with relatively higher molecular weights. Larger amounts of initiator also provide polymers with higher molecular weights. Higher pressure favors polymers with relatively higher molecular weights. The applications of these soy-based materials will be in the lubrication and hydraulic fluid areas.

  12. A Green Method for Processing Polymers using Dense Gas Technology

    PubMed Central

    Yoganathan, Roshan B.; Mammucari, Raffaella; Foster, Neil R.

    2010-01-01

    Dense CO2 can be used as an environmentally-benign polymer processing medium because of its liquid-like densities and gas-like mass transfer properties.In this work, polymer bio-blends of polycarbonate (PC), a biocompatible polymer, and polycaprolactone (PCL), a biodegradable polymer were prepared. Dense CO2 was used as a reaction medium for the melt-phase PC polymerization in the presence of dense CO2-swollen PCL particles and this method was used to prepare porous PC/PCL blends. To extend the applicability of dense CO2 to the biomedical industry and polymer blend processing, the impregnation of ibuprofen into the blend was conducted and subsequent dissolution characteristics were observed.

  13. Validation of Dissolution Testing with Biorelevant Media: An OrBiTo Study.

    PubMed

    Mann, James; Dressman, Jennifer; Rosenblatt, Karin; Ashworth, Lee; Muenster, Uwe; Frank, Kerstin; Hutchins, Paul; Williams, James; Klumpp, Lukas; Wielockx, Kristina; Berben, Philippe; Augustijns, Patrick; Holm, Rene; Hofmann, Michael; Patel, Sanjaykumar; Beato, Stefania; Ojala, Krista; Tomaszewska, Irena; Bruel, Jean-Luc; Butler, James

    2017-12-04

    Dissolution testing with biorelevant media has become widespread in the pharmaceutical industry as a means of better understanding how drugs and formulations behave in the gastrointestinal tract. Until now, however, there have been few attempts to gauge the reproducibility of results obtained with these methods. The aim of this study was to determine the interlaboratory reproducibility of biorelevant dissolution testing, using the paddle apparatus (USP 2). Thirteen industrial and three academic laboratories participated in this study. All laboratories were provided with standard protocols for running the tests: dissolution in FaSSGF to simulate release in the stomach, dissolution in a single intestinal medium, FaSSIF, to simulate release in the small intestine, and a "transfer" (two-stage) protocol to simulate the concentration profile when conditions are changed from the gastric to the intestinal environment. The test products chosen were commercially available ibuprofen tablets and zafirlukast tablets. The biorelevant dissolution tests showed a high degree of reproducibility among the participating laboratories, even though several different batches of the commercially available medium preparation powder were used. Likewise, results were almost identicalbetween the commercial biorelevant media and those produced in-house. Comparing results to previous ring studies, including those performed with USP calibrator tablets or commercially available pharmaceutical products in a single medium, the results for the biorelevant studies were highly reproducible on an interlaboratory basis. Interlaboratory reproducibility with the two-stage test was also acceptable, although the variability was somewhat greater than with the single medium tests. Biorelevant dissolution testing is highly reproducible among laboratories and can be relied upon for cross-laboratory comparisons.

  14. Assessment of International Students through the Medium of English: Ensuring Validity and Fairness in Content-Based Examinations

    ERIC Educational Resources Information Center

    Shaw, Stuart; Imam, Helen

    2013-01-01

    International assessments in a wide range of subjects are being prepared for and delivered through the medium of English in a variety of educational contexts. These assessments are taken by many candidates whose first language is not necessarily English. This raises important issues relating to assessment validity and fairness. This study…

  15. The Use of Financial Management Practices by Small, Medium and Micro Enterprises: A Perspective from South Africa

    ERIC Educational Resources Information Center

    Brijlal, Pradeep; Enow, Samuel; Isaacs, Eslyn B. H.

    2014-01-01

    This paper reports on an investigation of financial management practices used by small, medium-sized and micro-enterprises (SMMEs) in South Africa. It was found that more than half the SMMEs examined use external accounting staff to prepare accounting reports and more than 60% rely on external accounting staff to interpret and use accounting…

  16. High-frequency and hot-platen curing of medium-density fiberboards

    Treesearch

    R.R. Stevens; G.E. Woodson

    1977-01-01

    The effects of two curing methods- high-frequency heating and hot-platen heating- on the properties of a ure-formaldehyd-bonded medium-density fiberboard prepared with a southern-hardwoods furnish (50% southern red oak, 25% mockernut hickory, and 25% sweetgum) were studied. Boards of three densities- 38, 44, and 50 lb./ft.3- were cured by the two...

  17. Supercapacitors based on nitrogen-doped reduced graphene oxide and borocarbonitrides

    NASA Astrophysics Data System (ADS)

    Gopalakrishnan, K.; Moses, Kota; Govindaraj, A.; Rao, C. N. R.

    2013-12-01

    Nitrogen-doped reduced graphene oxide (RGO) samples with different nitrogen content, prepared by two different methods, as well as nitrogen-doped few-layer graphene have been investigated as supercapacitor electrodes. Two electrode measurements have been carried out both in aqueous (6M KOH) and in ionic liquid media. Nitrogen-doped reduced graphene oxides exhibit satisfactory specific capacitance, the values reaching 126F/g at a scan rate of 10mV/s in aqueous medium. Besides providing supercapacitor characteristics, the study has shown the nitrogen content and surface area to be important factors. High surface-area borocarbonitrides, BxCyNz, prepared by the urea route appear to be excellent supercapacitor electrode materials. Thus, BC4.5N exhibits a specific capacitance of 169F/g at a scan rate of 10mV/s in aqueous medium. In an ionic liquid medium, nitrogen-doped RGO and BC4.5N exhibit specific capacitance values of 258F/g and 240F/g at a scan rate of 5mV/s. The ionic liquid enables a larger operating voltage range of 0.0-2.5V compared to 0.0-1V in aqueous medium.

  18. Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures.

    PubMed

    Ukkonen, Kaisa; Veijola, Johanna; Vasala, Antti; Neubauer, Peter

    2013-07-29

    Fab antibody fragments in E. coli are usually directed to the oxidizing periplasmic space for correct folding. From periplasm Fab fragments may further leak into extracellular medium. Information on the cultivation parameters affecting this leakage is scarce, and the unpredictable nature of Fab leakage is problematic regarding consistent product recovery. To elucidate the effects of cultivation conditions, we investigated Fab expression and accumulation into either periplasm or medium in E. coli K-12 and E. coli BL21 when grown in different types of media and under different aeration conditions. Small-scale Fab expression demonstrated significant differences in yield and ratio of periplasmic to extracellular Fab between different culture media and host strains. Expression in a medium with fed-batch-like glucose feeding provided highest total and extracellular yields in both strains. Unexpectedly, cultivation in baffled shake flasks at 150 rpm shaking speed resulted in higher yield and accumulation of Fabs into culture medium as compared to cultivation at 250 rpm. In the fed-batch medium, extracellular fraction in E. coli K-12 increased from 2-17% of total Fab at 250 rpm up to 75% at 150 rpm. This was partly due to increased lysis, but also leakage from intact cells increased at the lower shaking speed. Total Fab yield in E. coli BL21 in glycerol-based autoinduction medium was 5 to 9-fold higher at the lower shaking speed, and the extracellular fraction increased from ≤ 10% to 20-90%. The effect of aeration on Fab localization was reproduced in multiwell plate by variation of culture volume. Yield and leakage of Fab fragments are dependent on expression strain, culture medium, aeration rate, and the combination of these parameters. Maximum productivity in fed-batch-like conditions and in autoinduction medium is achieved under sufficiently oxygen-limited conditions, and lower aeration also promotes increased Fab accumulation into extracellular medium. These findings have practical implications for screening applications and small-scale Fab production, and highlight the importance of maintaining consistent aeration conditions during scale-up to avoid changes in product yield and localization. On the other hand, the dependency of Fab leakage on cultivation conditions provides a practical way to manipulate Fab localization.

  19. Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures

    PubMed Central

    2013-01-01

    Background Fab antibody fragments in E. coli are usually directed to the oxidizing periplasmic space for correct folding. From periplasm Fab fragments may further leak into extracellular medium. Information on the cultivation parameters affecting this leakage is scarce, and the unpredictable nature of Fab leakage is problematic regarding consistent product recovery. To elucidate the effects of cultivation conditions, we investigated Fab expression and accumulation into either periplasm or medium in E. coli K-12 and E. coli BL21 when grown in different types of media and under different aeration conditions. Results Small-scale Fab expression demonstrated significant differences in yield and ratio of periplasmic to extracellular Fab between different culture media and host strains. Expression in a medium with fed-batch-like glucose feeding provided highest total and extracellular yields in both strains. Unexpectedly, cultivation in baffled shake flasks at 150 rpm shaking speed resulted in higher yield and accumulation of Fabs into culture medium as compared to cultivation at 250 rpm. In the fed-batch medium, extracellular fraction in E. coli K-12 increased from 2-17% of total Fab at 250 rpm up to 75% at 150 rpm. This was partly due to increased lysis, but also leakage from intact cells increased at the lower shaking speed. Total Fab yield in E. coli BL21 in glycerol-based autoinduction medium was 5 to 9-fold higher at the lower shaking speed, and the extracellular fraction increased from ≤ 10% to 20-90%. The effect of aeration on Fab localization was reproduced in multiwell plate by variation of culture volume. Conclusions Yield and leakage of Fab fragments are dependent on expression strain, culture medium, aeration rate, and the combination of these parameters. Maximum productivity in fed-batch-like conditions and in autoinduction medium is achieved under sufficiently oxygen-limited conditions, and lower aeration also promotes increased Fab accumulation into extracellular medium. These findings have practical implications for screening applications and small-scale Fab production, and highlight the importance of maintaining consistent aeration conditions during scale-up to avoid changes in product yield and localization. On the other hand, the dependency of Fab leakage on cultivation conditions provides a practical way to manipulate Fab localization. PMID:23895637

  20. Enhancing effect of medium-chain triglycerides on intestinal absorption of d-alpha-tocopherol acetate from lecithin-dispersed preparations in the rat.

    PubMed

    Fukui, E; Kurohara, H; Kageyu, A; Kurosaki, Y; Nakayama, T; Kimura, T

    1989-02-01

    The effect of formulations of lecithin-dispersed preparation on the absorption of d-alpha-tocopherol acetate (VEA) from the small intestine was investigated in rats. When lecithin-dispersed preparations containing VEA or polysorbate 80 (PS-80)-solubilized solution of VEA were intraduodenally administered, VEA was hydrolyzed to d-alpha-tocopherol (VE) and was not detected in the plasma nor in the thoracic lymph. The maximum plasma concentration (Cmax) of VE after the intraduodenal administration of a preparation consisting of VEA, soybean phosphatidylcholine (PC) and medium-chain triglycerides (MCTG) (VEA/PC/MCTG, 5/16/1 by weight) was highest among the VEA preparations, and PS-80-solubilized solution gave the lowest Cmax. AUC of VE up to 24 h was also increased by the addition of MCTG to VEA/PC preparation. In the thoracic duct-fistula rat, the transport of VE into the thoracic lymph was increased by the administration of the VEA/PC/MCTG preparation significantly more than the VEA/PC preparation; the cumulative amounts of VE recovered in the thoracic lymph up to 24 h were 23.2 +/- 0.5% and 10.9 +/- 1.5% of dose, respectively. The plasma concentration of VE was not increased in the thoracic duct-fistula rat even after the intraduodenal administration of VEA preparations, suggesting that VE is not transported directly to the systemic circulation, but by way of the lymphatic route. The lymphatic transport of VE following the intraduodenal administration of VEA/PC/MCTG preparation was markedly diminished by the simultaneous administration of Pluronic L-81 emulsion, an inhibitor of chylomicron formation. It is suggested that the chylomicron is essential to the lymphatic transport of VE from VEA preparations.

  1. Transition to surgical residency: a multi-institutional study of perceived intern preparedness and the effect of a formal residency preparatory course in the fourth year of medical school.

    PubMed

    Minter, Rebecca M; Amos, Keith D; Bentz, Michael L; Blair, Patrice Gabler; Brandt, Christopher; D'Cunha, Jonathan; Davis, Elisabeth; Delman, Keith A; Deutsch, Ellen S; Divino, Celia; Kingsley, Darra; Klingensmith, Mary; Meterissian, Sarkis; Sachdeva, Ajit K; Terhune, Kyla; Termuhlen, Paula M; Mullan, Patricia B

    2015-08-01

    To evaluate interns' perceived preparedness for defined surgical residency responsibilities and to determine whether fourth-year medical school (M4) preparatory courses ("bootcamps") facilitate transition to internship. The authors conducted a multi-institutional, mixed-methods study (June 2009) evaluating interns from 11 U.S. and Canadian surgery residency programs. Interns completed structured surveys and answered open-ended reflective questions about their preparedness for their surgery internship. Analyses include t tests comparing ratings of interns who had and had not participated in formal internship preparation programs. The authors calculated Cohen d for effect size and used grounded theory to identify themes in the interns' reflections. Of 221 eligible interns, 158 (71.5%) participated. Interns self-reported only moderate preparation for most defined care responsibilities in the medical knowledge and patient care domains but, overall, felt well prepared in the professionalism, interpersonal communication, practice-based learning, and systems-based practice domains. Interns who participated in M4 preparatory curricula had higher self-assessed ratings of surgical technical skills, professionalism, interpersonal communication skills, and overall preparation, at statistically significant levels (P < .05) with medium effect sizes. Themes identified in interns' characterizations of their greatest internship challenges included anxiety or lack of preparation related to performance of technical skills or procedures, managing simultaneous demands, being first responders for critically ill patients, clinical management of predictable postoperative conditions, and difficult communications. Entering surgical residency, interns report not feeling prepared to fulfill common clinical and professional responsibilities. As M4 curricula may enhance preparation, programs facilitating transition to residency should be developed and evaluated.

  2. English Medium Instruction: A Way towards Linguistically Better Prepared Professionals in the Basque Autonomous Community?

    ERIC Educational Resources Information Center

    van der Worp, Karin

    2017-01-01

    In the Basque Autonomous Community, besides the official languages Spanish and Basque, English is considered an important third language for internationally operating companies. However, employees are not believed to be linguistically well enough prepared, due to shortcomings in English language learning in the Basque educational system. The…

  3. Preparing International Counselor Educators through Online Distance Learning

    ERIC Educational Resources Information Center

    Sells, James; Tan, Augustine; Brogan, Justin; Dahlen, Ulla; Stupart, Yvette

    2012-01-01

    Online learning has emerged as a significant teaching medium, influencing how higher education is conducted in the U.S. and globally. While widely accepted in select disciplines, distance learning as the method for counselor preparation is often met with skepticism or resistance in the counseling profession. The purpose of this paper is to argue…

  4. Facile synthesis of TiO2/microcrystalline cellulose nanocomposites: photocatalytically active material under visible light irradiation

    EPA Science Inventory

    Doped TiO2 nanocomposites were prepared in situ by a facile and simple synthesis utilizing benign and renewable precursors such as microcrystalline cellulose (MC) and TiCl4 through hydrolysis in alkaline medium without the addition of organic solvents. The as-prepared nanocompos...

  5. Preparative Isolation and Purification of Flavone C-Glycosides from the Leaves of Ficus microcarpa L. f by Medium-Pressure Liquid Chromatography, High-Speed Countercurrent Chromatography, and Preparative Liquid Chromatography

    PubMed Central

    Wang, Xiaohong; Liang, Yong; Zhu, Licai; Xie, Huichun; Li, Hang; He, Junting; Pan, Man; Zhang, Tianyou; Ito, Yoichiro

    2009-01-01

    Combined with medium-pressure liquid chromatography (MPLC) and preparative high-performance liquid chromatography (perp-HPLC), high-speed countercurrent chromatography (HSCCC) was applied for separation and purification of flavone C-glycosides from the crude extract of leaves of Ficus microcarpae L. f. HSCCC separation was performed on a two-phase solvent system composed of methyl tert- butyl ether - ethyl acetate – 1-butanol – acetonitrile – 0.1% aqueous trifluoroacetic acid at a volume ratio of 1:3:1:1:5. Partially resolved peak fractions from HSCCC separation were further purified by preparative HPLC. Four well-separated compounds were obtained and their purities were determined by HPLC. The purities of these peaks were 97.28%, 97.20%, 92.23%, and 98.40%.. These peaks were characterized by ESI-MSn. According to the reference, they were identified as orientin (peak I), isovitexin-3″-O-glucopyranoside (peak II), isovitexin (peak III), and vitexin (peak IV), yielded 1.2 mg, 4.5 mg, 3.3 mg, and 1.8 mg, respectively. PMID:20190866

  6. Dissolution of Lipid-Based Matrices in Simulated Gastrointestinal Solutions to Evaluate Their Potential for the Encapsulation of Bioactive Ingredients for Foods.

    PubMed

    Raymond, Yves; Champagne, Claude P

    2014-01-01

    The goal of the study was to compare the dissolution of chocolate to other lipid-based matrices suitable for the microencapsulation of bioactive ingredients in simulated gastrointestinal solutions. Particles having approximately 750 μm or 2.5 mm were prepared from the following lipid-based matrices: cocoa butter, fractionated palm kernel oil (FPKO), chocolate, beeswax, carnauba wax, and paraffin. They were added to solutions designed to simulate gastric secretions (GS) or duodenum secretions (DS) at 37°C. Paraffin, carnauba wax, and bees wax did not dissolve in either the GS or DS media. Cocoa butter, FPKO, and chocolate dissolved in the DS medium. Cocoa butter, and to a lesser extent chocolate, also dissolved in the GS medium. With chocolate, dissolution was twice as fast as that with small particles (750 μm) as compared to the larger (2.5 mm) ones. With 750 μm particle sizes, 90% dissolution of chocolate beads was attained after only 60 minutes in the DS medium, while it took 120 minutes for 70% of FPKO beads to dissolve in the same conditions. The data are discussed from the perspective of controlled release in the gastrointestinal tract of encapsulated ingredients (minerals, oils, probiotic bacteria, enzymes, vitamins, and peptides) used in the development of functional foods.

  7. In vitro cultivation of anisakis simplex: pepsin increases survival and moulting from fourth larval to adult stage.

    PubMed

    Iglesias, L; Valero, A; Benítez, R; Adroher, F J

    2001-09-01

    This paper describes the in vitro cultivation of the 3rd-larval stage (L3) of Anisakis simplex to adulthood in a much simpler and easier to prepare medium than those described to date. The adult males obtained are between 3.8 and 6.5 cm long and the females between 4.5 and 8.0 cm. Some individually cultivated females laid eggs which had an average size of 44.4 x 50.5 microm. The culture conditions were as follows: medium RPMI-1640 supplemented with 20% heat-inactivated fetal bovine serum and 1% commercial pepsin, at pH 4.0 and a temperature of 37 degrees C, and in air atmosphere with 5% CO2. The pepsin was found to be the key to the success of the culture. The average survival of the worms in the culture increased from 50 to 88 days, due to the fact that the survival of the adults practically doubled (increasing by 1.9 times). Furthermore, the number of worms that completed the 4th moulting (M4) increased by 4.2 times, from 22.9 to 95.6%. This culture medium may facilitate, due to its simplicity, the study of anisakids, or at least of A. simplex, constituting another step towards achieving a complete in vitro life-cycle for these parasites.

  8. Dissolution of Lipid-Based Matrices in Simulated Gastrointestinal Solutions to Evaluate Their Potential for the Encapsulation of Bioactive Ingredients for Foods

    PubMed Central

    Champagne, Claude P.

    2014-01-01

    The goal of the study was to compare the dissolution of chocolate to other lipid-based matrices suitable for the microencapsulation of bioactive ingredients in simulated gastrointestinal solutions. Particles having approximately 750 μm or 2.5 mm were prepared from the following lipid-based matrices: cocoa butter, fractionated palm kernel oil (FPKO), chocolate, beeswax, carnauba wax, and paraffin. They were added to solutions designed to simulate gastric secretions (GS) or duodenum secretions (DS) at 37°C. Paraffin, carnauba wax, and bees wax did not dissolve in either the GS or DS media. Cocoa butter, FPKO, and chocolate dissolved in the DS medium. Cocoa butter, and to a lesser extent chocolate, also dissolved in the GS medium. With chocolate, dissolution was twice as fast as that with small particles (750 μm) as compared to the larger (2.5 mm) ones. With 750 μm particle sizes, 90% dissolution of chocolate beads was attained after only 60 minutes in the DS medium, while it took 120 minutes for 70% of FPKO beads to dissolve in the same conditions. The data are discussed from the perspective of controlled release in the gastrointestinal tract of encapsulated ingredients (minerals, oils, probiotic bacteria, enzymes, vitamins, and peptides) used in the development of functional foods. PMID:26904647

  9. Entanglement analysis of a two-atom nonlinear Jaynes-Cummings model with nondegenerate two-photon transition, Kerr nonlinearity, and two-mode Stark shift

    NASA Astrophysics Data System (ADS)

    Baghshahi, H. R.; Tavassoly, M. K.; Faghihi, M. J.

    2014-12-01

    An entangled state, as an essential tool in quantum information processing, may be generated through the interaction between light and matter in cavity quantum electrodynamics. In this paper, we study the interaction between two two-level atoms and a two-mode field in an optical cavity enclosed by a medium with Kerr nonlinearity in the presence of a detuning parameter and Stark effect. It is assumed that the atom-field coupling and third-order susceptibility of the Kerr medium depend on the intensity of the light. In order to investigate the dynamics of the introduced system, we obtain the exact analytical form of the state vector of the considered atom-field system under initial conditions which may be prepared for the atoms (in a coherent superposition of their ground and upper states) and the fields (in a standard coherent state). Then, in order to evaluate the degree of entanglement between the subsystems, we investigate the dynamics of the entanglement by employing the entanglement of formation. Finally, we analyze in detail the influences of the Stark shift, the deformed Kerr medium, the intensity-dependent coupling, and also the detuning parameter on the behavior of this measure for different subsystems. The numerical results show that the amount of entanglement between the different subsystems can be controlled by choosing the evolved parameters appropriately.

  10. Effect of environmental and cultural conditions on medium pH and explant growth performance of Douglas-fir ( Pseudotsuga menziesii) shoot cultures

    PubMed Central

    Chen, Chien-Chih; Bates, Rick; Carlson, John

    2015-01-01

    The medium pH level of plant tissue cultures has been shown to be essential to many aspects of explant development and growth. Sensitivity or tolerance of medium pH change in vitro varies according to specific requirements of individual species. The objectives of this study are to 1) determine medium pH change over time in storage conditions and with presence of explants, 2) evaluate the effects of medium pH change on explant growth performance and 3) assess the effects of adding a pH stabilizer, 2-(N-morpholino)ethanesulfonic acid (MES) that is commonly used in Douglas-fir micropropagation medium. Vegetative buds were collected in the spring before breaking dormancy from juvenile and mature donor trees for conducting these evaluations. Medium, with or without MES, was pre-adjusted to five pH levels before adding MES, agar and autoclaving. Medium pH changes and explant growth parameters were measured at eight different incubation times. Overall, MES provided a more stable medium pH, relative to starting pH values, under both light and dark storage conditions as well as with presence of explants. A general trend of decreasing medium pH over time was found comparing explants from juvenile and mature donor genotypes. Explant height and weight growth increased over time, but differ among explants from juvenile and mature donor genotypes. Our findings suggest that a 21-day subculture practice may best sustain medium freshness, medium pH level and desirable explant growth. PMID:26535110

  11. Antitumor Activity of Rat Mesenchymal Stem Cells during Direct or Indirect Co-Culturing with C6 Glioma Cells.

    PubMed

    Gabashvili, A N; Baklaushev, V P; Grinenko, N F; Mel'nikov, P A; Cherepanov, S A; Levinsky, A B; Chehonin, V P

    2016-02-01

    The tumor-suppressive effect of rat mesenchymal stem cells against low-differentiated rat C6 glioma cells during their direct and indirect co-culturing and during culturing of C6 glioma cells in the medium conditioned by mesenchymal stem cells was studied in an in vitro experiment. The most pronounced antitumor activity of mesenchymal stem cells was observed during direct co-culturing with C6 glioma cells. The number of live C6 glioma cells during indirect co-culturing and during culturing in conditioned medium was slightly higher than during direct co-culturing, but significantly differed from the control (C6 glioma cells cultured in medium conditioned by C6 glioma cells). The cytotoxic effect of medium conditioned by mesenchymal stem cells was not related to medium depletion by glioma cells during their growth. The medium conditioned by other "non-stem" cells (rat astrocytes and fibroblasts) produced no tumor-suppressive effect. Rat mesenchymal stem cells, similar to rat C6 glioma cells express connexin 43, the main astroglial gap junction protein. During co-culturing, mesenchymal stem cells and glioma C6 cells formed functionally active gap junctions. Gap junction blockade with connexon inhibitor carbenoxolone attenuated the antitumor effect observed during direct co-culturing of C6 glioma cells and mesenchymal stem cells to the level produced by conditioned medium. Cell-cell signaling mediated by gap junctions can be a mechanism of the tumor-suppressive effect of mesenchymal stem cells against C6 glioma cells. This phenomenon can be used for the development of new methods of cell therapy for high-grade malignant gliomas.

  12. Polyelectrolyte complexes between (cross-linked) N-carboxyethylchitosan and (quaternized) poly[2-(dimethylamino)ethyl methacrylate]: preparation, characterization, and antibacterial properties.

    PubMed

    Yancheva, Elena; Paneva, Dilyana; Maximova, Vera; Mespouille, Laetitia; Dubois, Philippe; Manolova, Nevena; Rashkov, Iliya

    2007-03-01

    Novel polyelectrolyte complexes (PECs) between N-carboxyethylchitosan (CECh) and well-defined (quaternized) poly[2-(dimethylamino)ethyl methacrylate] (PDMAEMA) have been obtained. The modification of chitosan into CECh allows the preparation of PECs in a pH range in which chitosan cannot form complexes. The CECh/PDMAEMA complex is formed in a narrow pH range around 7. The quaternization of the tertiary amino groups of PDMAEMA enables complex formation with CECh both in neutral and in alkaline medium. Cross-linked CECh is also capable of forming complexes with (quaternized) PDMAEMA. The antibacterial activity of (cross-linked) CECh, (quaternized) PDMAEMA, and their complexes against Escherichia coli has been evaluated. In contrast to (quaternized) PDMAEMA, (cross-linked) CECh exhibits no antibacterial activity. The complex formation between cross-linked CECh and (quaternized) PDMAEMA results in a loss of the inherent antibacterial activity of the latter in neutral medium. In acidic medium, the complexes exhibit strong antibacterial activity due to complex disintegration and release of (quaternized) PDMAEMA.

  13. Preparation of the Iron Oxide Red from the Converter Dust by the Magnetic Separation and Roasting Process

    NASA Astrophysics Data System (ADS)

    Guo, Z. J.; Li, S. Q.; Yang, C. Q.

    2017-05-01

    Preparation of iron oxide red (α-Fe2O3) from the converter dust by the superconductivity high gradient magnetic separation (S-HGMS) and roasting process was investigated in the paper. The basic properties of the dust were studied by the X Ray Fluorescence, the chemical analysis and the X Ray Diffraction methods. The results showed that the raw dust mainly contained elements of Fe‵O‵Si‵Ca, the iron content of the raw dust was 61.80%, and there were ferrous phases of Fe3O4, α-Fe2O3, Fe2(SiO4) and CaFe(Si2O6) in the raw dust. Under the optimum conditions of magnetic field intensity of 1.8T, the dispersion agent of 30mg/L and velocity of 500mL/min, the powders absorbed by the magnetic medium mainly contained Fe3O4 and α-Fe2O3, and the iron content of powders absorbed was up to 65.90%. The Fe2+ content of the powders absorbed under the optimum magnetic conditions dropped to 0.25% from 19.10% after roasting of fifty minutes, and the iron content of powders absorbed under the optimum magnetic conditions fell to 64% due to oxidation, and the Fe3O4 was removed. Finally the α-Fe2O3 content was up to 91.07% in the iron oxide red.

  14. Generation of an unusual depolarizing response in rabbit primary afferent neurones in the absence of divalent cations.

    PubMed

    Stansfeld, C E; Wallis, D I

    1984-07-01

    The effects of divalent cations on responses to 5-hydroxytryptamine (5-HT), gamma-aminobutyric acid (GABA) and 1,1-dimethyl-4-phenyl piperazinium (DMPP) were investigated using a sucrose-gap method to record population responses. In Ca-free medium responses to 5-HT were enhanced, those to DMPP depressed and those to GABA unchanged. In Mg-free medium responses to 5-HT were unchanged, while those to DMPP and GABA were depressed. Removal of both Ca and Mg from the superfusion medium caused a small reduction of GABA responses and a large reduction of DMPP responses. Responses to 5-HT were not only greatly potentiated but were changed in character; the depolarizing phase became sigmoid and the dose dependence between quantity of 5-HT and response magnitude was lost as if 5-HT were triggering an all-or-nothing phenomenon. Dose--response relationships for GABA were normal in the large majority of preparations. In about 10% of preparations, supramaximal amounts of GABA or DMPP evoked large responses of a similar character to those evoked by 5-HT. The large responses, generated by an unknown mechanism, were termed X responses. Further reduction in tissue divalent cations by EGTA (1 mM) caused X responses to be generated spontaneously. Ca, Mg, Mn or Co (1 mM) could suppress X responses. DMPP responses, reduced in Ca/Mg-free medium, were largely restored by 1 mM-Ca. Depression of GABA responses in Ca/Mg-free medium could be entirely attributed to the absence of Mg, Mn being able to substitute for Mg. X responses were generated only after equilibration for 1 h with Ca/Mg-free medium. Attempts to manipulate [Ca]i with dinitrophenol or caffeine did not produce the conditions under which X responses were generated. Intracellular records of responses to 5-HT, GABA or DMPP showed that cells with A fibres responded to GABA but not to 5-HT or DMPP. Fifty-four out of sixty-seven cells with C fibre axons (80%) were depolarized by 5-HT, thirty-seven out of forty-nine (76%) by DMPP and forty out of fifty-seven (70%) by GABA. Eighteen out of thirty-eight (47%) C cells were depolarized by all three agents. Some C cells were very sensitive to 5-HT, 10(-6) M evoking a substantial response. In most, responses to 10(-5) M-5-HT had a slower rate of rise than responses to 10(-4) or 10(-3) M-GABA or DMPP, yet lower 5-HT concentrations normally elicited X responses in sucrose-gap experiments whereas GABA or DMPP normally did not.(ABSTRACT TRUNCATED AT 400 WORDS)

  15. Novel biorelevant dissolution medium as a prognostic tool for polysaccharide-based colon-targeted drug delivery system.

    PubMed

    Yadav, Ankit Kumar; Sadora, Manik; Singh, Sachin Kumar; Gulati, Monica; Maharshi, Peddi; Sharma, Abhinav; Kumar, Bimlesh; Rathee, Harish; Ghai, Deepak; Malik, Adil Hussain; Garg, Varun; Gowthamrajan, K

    2017-01-01

    To overcome the limitations of the conventionally used methods for evaluation of orally administered colon-targeted delivery systems, a novel dissolution method using probiotics has been recently reported. In the present study, universal suitability of this medium composed of five different probiotics is established. Different delivery systems - mini tablets, liquisolid compacts, and microspheres coated with different polysaccharides - were prepared and subjected to sequential dissolution testing in medium with and without microbiota. The results obtained from fluid thioglycollate medium (FTM)-based probiotic medium for all the polysaccharide-based formulations showed statistically similar dissolution profile to that in the rat and goat cecal content media. Hence, it can be concluded that the developed FTM-based probiotic medium, once established, may eliminate the need for further animal sacrifice in the dissolution testing of polysaccharide-based colon-targeted delivery system.

  16. Thin stillage supplementation greatly enhances bacterial cellulose production by Gluconacetobacter xylinus.

    PubMed

    Wu, Jyh-Ming; Liu, Ren-Han

    2012-09-01

    Thin stillage (TS), a wastewater from rice wine distillery can well sustain the growth of Gluconacetobacter xylinus for production of bacterial cellulose (BC). When used as a supplement to the traditional BC production medium (Hestrin and Schramm medium), the enhancement of BC production increased with the amount of TS supplemented in a static culture of G. xylinus. When TS was employed to replace distilled water for preparing HS medium (100%TS-HS medium), the BC production in this 100%TS-HS medium was enhanced 2.5-fold to a concentration of 10.38 g/l with sugar to BC conversion yield of 57% after 7 days cultivation. The cost-free TS as a supplement in BC production medium not only can greatly enhance the BC production, but also can effectively dispose the nuisance wastewater of rice wine distillery. Copyright © 2012 Elsevier Ltd. All rights reserved.

  17. X-irradiation of human bronchial cancer cells causes the bystander effects in normal bronchial cells in vitro.

    PubMed

    Konopacka, M; Rogoliński, J

    2010-01-01

    Using X radiation commonly used in radiotherapy of cancers we investigated bystander interactions between human cells: irradiated A549 bronchial carcinoma human cells and non irradiated BEAS-2B normal bronchial epithelial cells. Non irradiated cells were incubated in medium transferred from irradiated A549 cells (ICM-irradiation conditioned medium) for 48h and next the chromosomal damage and apoptosis were estimated. Conditioned medium collected from irradiated cancer cells induced in non irradiated cells of the same line as well as in BEAS-2B normal cells genetic changes such as micronuclei, chromatid and chromosomal breaks and condensation of chromatin characteristic for processes of apoptosis. Addition of only 1% of conditioned medium to fresh medium was sufficient to induction of bystander response to normal bronchial cells. The presented results in this study could have implications for human radiation risk and in evaluating the secondary effects of radiotherapy.

  18. An invisible medium for circularly polarized electromagnetic waves.

    PubMed

    Tamayama, Y; Nakanishi, T; Sugiyama, K; Kitano, M

    2008-12-08

    We study the no reflection condition for a planar boundary between vacuum and an isotropic chiral medium. In general chiral media, elliptically polarized waves incident at a particular angle satisfy the no reflection condition. When the wave impedance and wavenumber of the chiral medium are equal to the corresponding parameters of vacuum, one of the circularly polarized waves is transmitted to the medium without reflection or refraction for all angles of incidence. We propose a circular polarizing beam splitter as a simple application of the no reflection effect. (c) 2008 Optical Society of America

  19. Radiochemistry, pre-clinical studies and first clinical investigation of 90Y-labeled hydroxyapatite (HA) particles prepared utilizing 90Y produced by (n,γ) route.

    PubMed

    Vimalnath, K V; Chakraborty, Sudipta; Rajeswari, A; Sarma, H D; Nuwad, Jitendra; Pandey, Usha; Kamaleshwaran, K; Shinto, Ajit; Dash, Ashutosh

    2015-05-01

    The scope of using no carrier added (NCA) (90)Y [T(1/2) = 64.1 h, Eβ(max) = 2.28 MeV] obtained from (90)Sr/(90)Y generator in radiation synovectomy (RSV) is widely accepted. In the present study, the prospect of using (90)Y produced by (n,γ) route in a medium flux research reactor for use in RSV was explored. Yttrium-90 was produced by thermal neutron irradiation of Y(2)O(3) target at a neutron flux of ~1×10(14) n/cm(2).s for 14 d. The influence of various experimental parameters were systematically investigated and optimized to arrive at the most favorable conditions for the formulation of (90)Y labeled hydroxyapatite (HA) using HA particles of 1-10 μm size range. An optimized kit formulation strategy was developed for convenient one-step compounding of (90)Y-HA, which is easily adaptable at hospital radiopharmacy. The pre-clinical biological evaluation of (90)Y-HA particles was studied by carrying out biodistribution and bioluminiscence imaging studies in Wistar rats. The first clinical investigation using the radiolabeled preparation was performed on a patient suffering from chronic arthritis in knee joint by administering 185 MBq (90)Y-HA formulated at the hospital radiopharmacy deploying the proposed strategy. Yttrium-90 was produced with a specific activity of 851 ± 111 MBq/mg and radionuclidic purity of 99.95 ± 0.02%. (90)Y-labeled HA particles (185 ± 10 MBq doses) were formulated in high radiochemical purity (>99%) and excellent in vitro stability. The preparation showed promising results in pre-clinical studies carried out in Wistar rats. The preliminary results of the first clinical investigation of (90)Y-HA preparation in a patient with rheumatoid arthritis in knee joints demonstrated the effectiveness of the formulation prepared using (90)Y produced via (n,γ) route in the management of the disease. The studies revealed that effective utilization of (90)Y produced via (n,γ) route in a medium flux research reactor coupled with the developed strategy of using HA kits for convenient formulation of (90)Y-HA at the hospital radiopharmacy can contribute to sustainable growth in the clinical utilization of (90)Y in RSV in the foreseeable future. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Large-area few-layer hexagonal boron nitride prepared by quadrupole field aided exfoliation

    NASA Astrophysics Data System (ADS)

    Lun Lu, Han; Zhi Rong, Min; Qiu Zhang, Ming

    2018-03-01

    A quadrupole electric field-mediated exfoliation method is proposed to convert micron-sized hexagonal boron nitride (h-BN) powder into few-layer hexagonal boron nitride nanosheets (h-BNNS). Under optimum conditions (400 Hz, 40 V, 32 μg ml-1, sodium deoxycholate, TAE medium), the h-BN powders (thickness >200 nm, horizontal scale ˜10 μm) are successfully exfoliated into 0.5-4 nm (1-10 layers) thick h-BNNS with the same horizontal scale. Dynamic laser scattering and atomic force microscope data show that the yield is 47.6% (for the portion with the thickness of 0.5-6 nm), and all of the vertical sizes are reduced to smaller than 18 nm (45 layers).

  1. New milk medium for germ tube and chlamydoconidia production by Candida albicans.

    PubMed

    Jitsurong, S; Kiamsiri, S; Pattararangrong, N

    1993-08-01

    A new medium consisting of UHT milk, tween 80 and agar is described for the development of both germ tube and chlamydoconidia by Candida albicans. In total 172 isolates from clinical specimens, including C. albicans (112), C. guilliermondii (4), C. krusei (3), C. parasilopsis (16). C. tropicalis (28), Torulopsis glabrata (6) and Trichosporon beigellii (3), were examined in this medium by using the standard method. A higher percentage (98.2%) of germ tube production by C. albicans was found in this medium than in undiluted serum (90.2%). In addition, only C. albicans was found to be able to produce a high percentage of chlamydoconidia (95.5%) after 48 hours' incubation. In comparison with the conventional medium, corn meal tween 80 agar (21.4%), this new medium gives a significantly higher percentage and abundance of chlamydoconidia production. Being simple, cheap and easy to prepare, the new milk medium is proposed as very practical in the clinical mycology laboratory.

  2. Reversible gelling culture media for in-vitro cell culture in three-dimensional matrices

    DOEpatents

    An, Yuehuei H.; Mironov, Vladimir A.; Gutowska, Anna

    2000-01-01

    A gelling cell culture medium useful for forming a three dimensional matrix for cell culture in vitro is prepared by copolymerizing an acrylamide derivative with a hydrophilic comonomer to form a reversible (preferably thermally reversible) gelling linear random copolymer in the form of a plurality of linear chains having a plurality of molecular weights greater than or equal to a minimum gelling molecular weight cutoff, mixing the copolymer with an aqueous solvent to form a reversible gelling solution and adding a cell culture medium to the gelling solution to form the gelling cell culture medium. Cells such as chondrocytes or hepatocytes are added to the culture medium to form a seeded culture medium, and temperature of the medium is raised to gel the seeded culture medium and form a three dimensional matrix containing the cells. After propagating the cells in the matrix, the cells may be recovered by lowering the temperature to dissolve the matrix and centrifuging.

  3. Preparation of stable Pickering emulsions with short, medium and long chain fats and starch nanocrystals and their in vitro digestion properties

    USDA-ARS?s Scientific Manuscript database

    Pickering emulsions are receiving more attention as delivery systems in food and pharmaceuticals because they can be formulated with nontoxic food ingredients to form stable emulsions. In this study, 40-100 nm starch nanocrystals (SNCs) prepared from acid hydrolysis of waxy maize starches were used ...

  4. Microbiological safety of commercial prime rib preparation methods: thermal inactivation of Salmonella spp. in mechanically tenderized beef roasts

    USDA-ARS?s Scientific Manuscript database

    A survey of food service operations in a medium-size Midwestern city was conducted to evaluate the microbiological safety of prime rib preparation methods relative to survival of Salmonella spp. in both intact and tenderized (non-intact) product. All six restaurants visited seared rib eye roasts (ai...

  5. Cyclopropyl glycine and proline-containing preparation noopept evoke two types of membrane potential responses in synaptoneurosomes.

    PubMed

    Lutsenko, V K; Vukolova, M N; Gudasheva, T A

    2003-06-01

    Proline, cyclo(Pro-Gly), and acyl-prolyl-containing dipeptide GVS-111 decreased synaptoneurosome membrane potential in a Ca2+-free medium. The efficiency of these preparations decreased in the following order: GVS>cyclo(Pro-Gly)>proline. Depolarization responses induced by endogenous nootropic agent cyclo(Pro-Gly) was dose-dependent and saturable; the threshold concentration of cyclo(Pro-Gly) was 10(-9) M. In a Ca2+-containing medium GVS and cyclo(Pro-Gly) induced both hyperpolarizing and depolarizing membrane responses of synaptoneurosomes. Possible mechanisms underlying changes in the membrane potential of synaptoneurosomes induced by nootropic agents are discussed. It was interesting whether modulation of electrogenesis can improve memory and potentiate the neuroprotective effect of the test nootropic agents.

  6. Preparation of fatty acid methyl esters for gas-chromatographic analysis of marine lipids: insight studies.

    PubMed

    Carvalho, Ana P; Malcata, F Xavier

    2005-06-29

    Assays for fatty acid composition in biological materials are commonly carried out by gas chromatography, after conversion of the lipid material into the corresponding methyl esters (FAME) via suitable derivatization reactions. Quantitative derivatization depends on the type of catalyst and processing conditions employed, as well as the solubility of said sample in the reaction medium. Most literature pertinent to derivatization has focused on differential comparison between alternative methods; although useful to find out the best method for a particular sample, additional studies on factors that may affect each step of FAME preparation are urged. In this work, the influence of various parameters in each step of derivatization reactions was studied, using both cod liver oil and microalgal biomass as model systems. The accuracies of said methodologies were tested via comparison with the AOCS standard method, whereas their reproducibility was assessed by analysis of variance of (replicated) data. Alkaline catalysts generated lower levels of long-chain unsaturated FAME than acidic ones. Among these, acetyl chloride and BF(3) were statistically equivalent to each other. The standard method, which involves alkaline treatment of samples before acidic methylation with BF(3), provided equivalent results when compared with acidic methylation with BF(3) alone. Polarity of the reaction medium was found to be of the utmost importance in the process: intermediate values of polarity [e.g., obtained by a 1:1 (v/v) mixture of methanol with diethyl ether or toluene] provided amounts of extracted polyunsaturated fatty acids statistically higher than those obtained via the standard method.

  7. Investigation of in situ gelling alginate formulations as a sustained release vehicle for co-precipitates of dextromethrophan and Eudragit S 100.

    PubMed

    El Maghraby, Gamal Mohamed; Elzayat, Ehab Mostafa; Alanazi, Fars Kaed

    2014-03-01

    Alginate vehicles are capable of forming a gel matrix in situ when they come into contact with gastric medium in the presence of calcium ions. However, the gel structure is pH dependent and can break after gastric emptying, leading to dose dumping. The aim of this work was to develop modified in situ gelling alginate formulations capable of sustaining dextromethorphan release throughout the gastrointestinal tract. Alginate solution (2 %, m/m) was used as a vehicle for the tested formulations. Solid matrix of the drug and Eudragit S 100 was prepared by dissolving the drug and polymer in acetone. The organic solvent was then evaporated and the deposited solid matrix was micronized, sieved and dispersed in alginate solution to obtain candidate formulations. The release behavior of dextromethorphan was monitored and evaluated in a medium simulating the gastric and intestinal pH. Drug-polymer compatibility and possible solid-state interactions suggested physical interaction through hydrogen bonding between the drug and the polymer. A significant decrease in the rate and extent of dextromethorphan release was observed with increasing Eudragit S 100 concentration in the prepared particles. Most formulations showed sustained release profiles similar to that of a commercial sustained-release liquid based on ion exchange resin. The release pattern indicated strict control of drug release both under gastric and intestinal conditions, suggesting the potential advantage of using a solid dispersion of drug-Eudragit S 100 to overcome the problem of dose dumping after the rupture of the pH dependent alginate gels.

  8. Human periodontal ligament stem cells secretome from multiple sclerosis patients suppresses NALP3 inflammasome activation in experimental autoimmune encephalomyelitis

    PubMed Central

    Soundara Rajan, Thangavelu; Giacoppo, Sabrina; Diomede, Francesca; Bramanti, Placido; Trubiani, Oriana; Mazzon, Emanuela

    2017-01-01

    Research in recent years has largely explored the immunomodulatory effects of mesenchymal stem cells (MSCs) and their secretory products, called “secretome,” in the treatment of neuroinflammatory diseases. Here, we examined whether such immunosuppressive effects might be elicited due to inflammasome inactivation. To this end, we treated experimental autoimmune encephalomyelitis (EAE) mice model of multiple sclerosis (MS) with the conditioned medium or purified exosomes/microvesicles (EMVs) obtained from relapsing-remitting-MS patients human periodontal ligament stem cells (hPDLSCs) and investigated the regulation of NALP3 inflammasome. We noticed enhanced expression of NALP3, Cleaved Caspase 1, interleukin (IL)-1β, and IL-18 in EAE mouse spinal cord. Conversely, hPDLSCs-conditioned medium and EMVs significantly blocked NALP3 inflammasome activation and provided protection from EAE. Reduction in NALP3, Cleaved Caspase 1, IL-1β, and IL-18 level was noticed in conditioned medium and EMVs-treated EAE mice. Pro-inflammatory Toll-like receptor (TLR)-4 and nuclear factor (NF)-κB were elevated in EAE, while hPDLSCs-conditioned medium and EMVs treatment reduced their expression and increased IκB-α expression. Characterization of hPDLSCs-conditioned medium showed substantial level of anti-inflammatory IL-10, transforming growth factor (TGF)-β, and stromal cell–derived factor 1α (SDF-1α). We propose that the immunosuppressive role of hPDLSCs-derived conditioned medium and EMVs in EAE mice may partly attribute to the presence of soluble immunomodulatory factors, NALP3 inflammasome inactivation, and NF-κB reduction. PMID:28764573

  9. Encapsulated microsensors for reservoir interrogation

    DOEpatents

    Scott, Eddie Elmer; Aines, Roger D.; Spadaccini, Christopher M.

    2016-03-08

    In one general embodiment, a system includes at least one microsensor configured to detect one or more conditions of a fluidic medium of a reservoir; and a receptacle, wherein the receptacle encapsulates the at least one microsensor. In another general embodiment, a method include injecting the encapsulated at least one microsensor as recited above into a fluidic medium of a reservoir; and detecting one or more conditions of the fluidic medium of the reservoir.

  10. Astrocytes Promote Oligodendrogenesis after White Matter Damage via Brain-Derived Neurotrophic Factor.

    PubMed

    Miyamoto, Nobukazu; Maki, Takakuni; Shindo, Akihiro; Liang, Anna C; Maeda, Mitsuyo; Egawa, Naohiro; Itoh, Kanako; Lo, Evan K; Lok, Josephine; Ihara, Masafumi; Arai, Ken

    2015-10-14

    Oligodendrocyte precursor cells (OPCs) in the adult brain contribute to white matter homeostasis. After white matter damage, OPCs compensate for oligodendrocyte loss by differentiating into mature oligodendrocytes. However, the underlying mechanisms remain to be fully defined. Here, we test the hypothesis that, during endogenous recovery from white matter ischemic injury, astrocytes support the maturation of OPCs by secreting brain-derived neurotrophic factor (BDNF). For in vitro experiments, cultured primary OPCs and astrocytes were prepared from postnatal day 2 rat cortex. When OPCs were subjected to chemical hypoxic stress by exposing them to sublethal CoCl2 for 7 d, in vitro OPC differentiation into oligodendrocytes was significantly suppressed. Conditioned medium from astrocytes (astro-medium) restored the process of OPC maturation even under the stressed conditions. When astro-medium was filtered with TrkB-Fc to remove BDNF, the BDNF-deficient astro-medium no longer supported OPC maturation. For in vivo experiments, we analyzed a transgenic mouse line (GFAP(cre)/BDNF(wt/fl)) in which BDNF expression is downregulated specifically in GFAP(+) astrocytes. Both wild-type (GFAP(wt)/BDNF(wt/fl) mice) and transgenic mice were subjected to prolonged cerebral hypoperfusion by bilateral common carotid artery stenosis. As expected, compared with wild-type mice, the transgenic mice exhibited a lower number of newly generated oligodendrocytes and larger white matter damage. Together, these findings demonstrate that, during endogenous recovery from white matter damage, astrocytes may promote oligodendrogenesis by secreting BDNF. The repair of white matter after brain injury and neurodegeneration remains a tremendous hurdle for a wide spectrum of CNS disorders. One potentially important opportunity may reside in the response of residual oligodendrocyte precursor cells (OPCs). OPCs may serve as a back-up for generating mature oligodendrocytes in damaged white matter. However, the underlying mechanisms are still mostly unknown. Here, we use a combination of cell biology and an animal model to report a new pathway in which astrocyte-derived BDNF supports oligodendrogenesis and regeneration after white matter damage. These findings provide new mechanistic insight into white matter physiology and pathophysiology, which would be broadly and clinically applicable to CNS disease. Copyright © 2015 the authors 0270-6474/15/3514002-07$15.00/0.

  11. Phenotyping the quality of complex medium components by simple online-monitored shake flask experiments.

    PubMed

    Diederichs, Sylvia; Korona, Anna; Staaden, Antje; Kroutil, Wolfgang; Honda, Kohsuke; Ohtake, Hisao; Büchs, Jochen

    2014-11-07

    Media containing yeast extracts and other complex raw materials are widely used for the cultivation of microorganisms. However, variations in the specific nutrient composition can occur, due to differences in the complex raw material ingredients and in the production of these components. These lot-to-lot variations can affect growth rate, product yield and product quality in laboratory investigations and biopharmaceutical production processes. In the FDA's Process Analytical Technology (PAT) initiative, the control and assessment of the quality of critical raw materials is one key aspect to maintain product quality and consistency. In this study, the Respiration Activity Monitoring System (RAMOS) was used to evaluate the impact of different yeast extracts and commercial complex auto-induction medium lots on metabolic activity and product yield of four recombinant Escherichia coli variants encoding different enzymes. Under non-induced conditions, the oxygen transfer rate (OTR) of E. coli was not affected by a variation of the supplemented yeast extract lot. The comparison of E. coli cultivations under induced conditions exhibited tremendous differences in OTR profiles and volumetric activity for all investigated yeast extract lots of different suppliers as well as lots of the same supplier independent of the E. coli variant. Cultivation in the commercial auto-induction medium lots revealed the same reproducible variations. In cultivations with parallel offline analysis, the highest volumetric activity was found at different cultivation times. Only by online monitoring of the cultures, a distinct cultivation phase (e.g. glycerol depletion) could be detected and chosen for comparable and reproducible offline analysis of the yield of functional product. This work proves that cultivations conducted in complex media may be prone to significant variation in final product quality and quantity if the quality of the raw material for medium preparation is not thoroughly checked. In this study, the RAMOS technique enabled a reliable and reproducible screening and phenotyping of complex raw material lots by online measurement of the respiration activity. Consequently, complex raw material lots can efficiently be assessed if the distinct effects on culture behavior and final product quality and quantity are visualized.

  12. Algal autolysate medium to label proteins for NMR in mammalian cells.

    PubMed

    Fuccio, Carmelo; Luchinat, Enrico; Barbieri, Letizia; Neri, Sara; Fragai, Marco

    2016-04-01

    In-cell NMR provides structural and functional information on proteins directly inside living cells. At present, the high costs of the labeled media for mammalian cells represent a limiting factor for the development of this methodology. Here we report a protocol to prepare a homemade growth medium from Spirulina platensis autolysate, suitable to express uniformly labeled proteins inside mammalian cells at a reduced cost-per-sample. The human proteins SOD1 and Mia40 were overexpressed in human cells grown in (15)N-enriched S. platensis algal-derived medium, and high quality in-cell NMR spectra were obtained.

  13. Enantioselective Synthesis of Medium-Sized Lactams via Chiral α,β-Unsaturated Acylammonium Salts.

    PubMed

    Kang, Guowei; Yamagami, Masaki; Vellalath, Sreekumar; Romo, Daniel

    2018-04-06

    Medium-sized lactams are important structural motifs found in a variety of bioactive compounds and natural products but are challenging to prepare, especially in optically active form. A Michael addition/proton transfer/lactamization organocascade process is described that delivers medium-sized lactams, including azepanones, benzazepinones, azocanones, and benzazocinones, in high enantiopurity through the intermediacy of chiral α,β-unsaturated acylammonium salts. An unexpected indoline synthesis was also uncovered, and the benzazocinone skeleton was transformed into other complex heterocyclic derivatives, including spiroglutarimides, isoquinolinones, and δ-lactones. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Recent advances in f-element separations based on a new method for the production of pentavalent americium in acidic solution

    DOE PAGES

    Mincher, Bruce J.; Schmitt, Nicholas C.; Schuetz, Brian K.; ...

    2015-03-11

    The peroxydisulfate anion has long been used for the preparation of hexavalent americium (Am VI) from the normally stable Am III valence state in mildly acidic solutions. However, there has been no satisfactory means to directly prepare the pentavalent state (Am V) in that medium. Some early literature reports indicated that the peroxydisulfate oxidation was incomplete, and silver ion catalysis in conjunction with peroxydisulfate became accepted as the means to ensure quantitative generation of Am VI. Incomplete oxidation would be expected to leave residual Am III, or to produce Am V in treated solutions. However, until recently, the use ofmore » peroxydisulfate as an Am V reagent has not been reported. Here, parameters influencing the oxidation were investigated, including peroxydisulfate and acid concentration, temperature, duration of oxidative treatment, and the presence of higher concentrations of other redox active metals such as plutonium. Using optimized conditions determined here, quantitative Am V was prepared in an acidic solution and the UV/Vis extinction coefficients of the Am V 513 nm peak were measured over a range of nitric acid concentrations. Furthermore, the utility of Am V for separations from the lanthanides and curium by solvent extraction, organic column chromatography and inorganic ion exchangers was also investigated.« less

  15. Preparation and characterization of triclosan nanoparticles for periodontal treatment.

    PubMed

    Piñón-Segundo, E; Ganem-Quintanar, A; Alonso-Pérez, V; Quintanar-Guerrero, D

    2005-04-27

    The aim of this work was to produce and characterize triclosan-loaded nanoparticles (NPs) by the emulsification-diffusion process, in an attempt to obtain a novel delivery system adequate for the treatment of periodontal disease. The NPs were prepared using poly(D,L-lactide-co-glycolide) (PLGA), poly(D,L-lactide) (PLA) and cellulose acetate phthalate (CAP). Poly(vinyl alcohol) (PVAL) was used as stabilizer. Batches were prepared with different amounts of triclosan (TCS) in order to evaluate the influence of drug on NP properties. Solid NPs of less than 500 nm in diameter were obtained. Entrapment efficiencies were higher than 63.8%. The characterization by scanning electron microscopy and light scattering indicated that high concentrations of TCS seemingly caused the increase of NP mean size. A decrease in the PLGA glass transition temperature was observed by differential scanning calorimetry. This could indicate that TCS in PLGA-NPs behaves as a non-conventional plasticizer. Subsequently, in vitro release studies were carried out under sink conditions using a device designed in our laboratory to allow a direct contact between the particles and the dissolution medium. A fast release of TCS from NPs was detected. A preliminary in vivo study in dogs with induced periodontal defects suggested that TCS-loaded NPs penetrate through the junctional epithelium.

  16. Germ tube and chlamydospore formation by Candida albicans on a new medium.

    PubMed

    Beheshti, F; Smith, A G; Krause, G W

    1975-10-01

    A new medium composed of "cream of rice" infusion, oxgall, Tween 80, and agar is described for the sequential development of germ tubes and chlamydospores by Candida albicans. The procedure used (Dalmau's technique) is an improvement over the fluid substrate procedures previously advocated for germ tube formation. That the same preparation is then used for chlamydospore production is of practical importance for the clinical mycology laboratory.

  17. Liquid chromatographic extraction medium

    DOEpatents

    Horwitz, E. Philip; Dietz, Mark L.

    1994-01-01

    A method and apparatus for extracting strontium and technetium values from biological, industrial and environmental sample solutions using a chromatographic column is described. An extractant medium for the column is prepared by generating a solution of a diluent containing a Crown ether and dispersing the solution on a resin substrate material. The sample solution is highly acidic and is introduced directed to the chromatographic column and strontium or technetium is eluted using deionized water.

  18. Battery Ownership Model - Medium Duty HEV Battery Leasing & Standardization

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

    Kelly, Ken; Smith, Kandler; Cosgrove, Jon

    2015-12-01

    Prepared for the U.S. Department of Energy, this milestone report focuses on the economics of leasing versus owning batteries for medium-duty hybrid electric vehicles as well as various battery standardization scenarios. The work described in this report was performed by members of the Energy Storage Team and the Vehicle Simulation Team in NREL's Transportation and Hydrogen Systems Center along with members of the Vehicles Analysis Team at Ricardo.

  19. Nanoparticle-supported and magnetically recoverable ruthenium hydroxide catalyst: efficient hydration of nitriles to amides in aqueous medium.

    PubMed

    Polshettiwar, Vivek; Varma, Rajender S

    2009-01-01

    Magnetic attraction not filtration: A magnetic nanoparticle-supported ruthenium hydroxide catalyst (see figure) was readily prepared from inexpensive starting materials and shown to catalyze the hydration of nitriles with excellent yield in a benign aqueous medium. Catalyst recovery using an external magnetic field, superior activity, and the inherent stability of the catalyst system are additional sustainable attributes of this protocol.

  20. Organic containment separator

    DOEpatents

    Del Mar, Peter

    1995-01-01

    A process of sample preparation prior to analysis for the concentration of an organic contaminant in an aqueous medium by (a) passing an initial aqueous medium including a minor amount of the organic contaminant through a composite tube including a polymeric base material selected from the group of polyolefins and polyfluorocarbons and particles of a carbon allotrope material adfixed to the inner wall of the polymeric base material, the composite tube having an internal diameter of from about 0.1 to about 2.0 millimeters and being of sufficient length to permit the organic contaminant to adhere to the composite tube, (b) passing a solvent through the composite tube, said solvent capable of separating the adhered organic contaminant from the composite tube. Further, an extraction apparatus for sample preparation prior to analysis for the concentration of an organic contaminant in an aqueous medium, said apparatus including a composite tube including a polymeric base material selected from the group of polyolefins and polyfluorocarbons and particles of a carbon allotrope material adfixed to the inner wall of the polymeric base material, the composite tube having an internal diameter of from about 0.1 to about 2.0 millimeters and being of sufficient length to permit an organic contaminant contained within an aqueous medium passed therethrough to adhere to the composite tube is disclosed.

  1. Organic contaminant separator

    DOEpatents

    Mar, Peter D.

    1994-01-01

    A process of sample preparation prior to analysis for the concentration of an organic contaminant in an aqueous medium by (a) passing an initial aqueous medium including a minor amount of the organic contaminant through a composite tube including a polymeric base material selected from the group of polyolefins and polyfluorocarbons and particles of a carbon allotrope material adfixed to the inner wall of the polymeric base material, the composite tube having an internal diameter of from about 0.1 to about 2.0 millimeters and being of sufficient length to permit the organic contaminant to adhere to the composite tube, (b) passing a solvent through the composite tube, said solvent capable of separating the adhered organic contaminant from the composite tube. Further, an extraction apparatus for sample preparation prior to analysis for the concentration of an organic contaminant in an aqueous medium, said apparatus including a composite tube including a polymeric base material selected from the group of polyolefins and polyfluorocarbons and particles of a carbon allotrope material adfixed to the inner wall of the polymeric base material, the composite tube having an internal diameter of from about 0.1 to about 2.0 millimeters and being of sufficient length to permit an organic contaminant contained within an aqueous medium passed therethrough to adhere to the composite tube is disclosed.

  2. Isolation and characterization of L-valine-degrading Candida maltosa DLPU-zpb for D-valine preparation from DL-valine.

    PubMed

    Zhang, C H; Xin, W T; Chen, M; Bi, Y; Gao, Z Q; Zhang, J

    2015-11-01

    To develop a practical process for D-valine preparation from DL-valine, L-valine was used as a sole source of carbon and nitrogen in basal minimal medium to isolate L-valine-degrading micro-organisms. A yeast strain DLPU-zpb was obtained, which showed asymmetric degrading activity against DL-valine. Based on the morphology, physiological and biochemical characteristics, and 26S rDNA D1/D2 domain sequence, strain DLPU-zpb was identified as Candida maltosa. The cells of this strain were used as a biocatalyst for eliminating the L-isomer from DL-valine. The L-isomer was completely degraded within 72 h under the conditions of 30°C, pH control at 6·0, 200 rev min(-1) and 50 g l(-1) DL-valine. The strain DLPU-zpb degraded L-valine effectively but not D-valine, and thus D-valine could be easily isolated from the resultant reaction mixture, which provides a new method for D-valine preparation from DL-valine. D-valine is an important raw material for medicines and its demand is increasing year by year. Several approaches for D-valine preparation have been reported, but none of them are likely to provide product at low cost. A newly isolated L-valine-degrading yeast strain Candida maltosa DLPU-zpb was described, which showed asymmetric degrading activity against DL-valine. Thus, a new and practical process for D-valine preparation from DL-valine could be developed. This is the first report of the asymmetric degrading ability of C. maltosa against DL-valine and D-valine preparation from DL-valine. © 2015 The Society for Applied Microbiology.

  3. Brain stem slice conditioned medium contains endogenous BDNF and GDNF that affect neural crest boundary cap cells in co-culture.

    PubMed

    Kaiser, Andreas; Kale, Ajay; Novozhilova, Ekaterina; Siratirakun, Piyaporn; Aquino, Jorge B; Thonabulsombat, Charoensri; Ernfors, Patrik; Olivius, Petri

    2014-05-30

    Conditioned medium (CM), made by collecting medium after a few days in cell culture and then re-using it to further stimulate other cells, is a known experimental concept since the 1950s. Our group has explored this technique to stimulate the performance of cells in culture in general, and to evaluate stem- and progenitor cell aptitude for auditory nerve repair enhancement in particular. As compared to other mediums, all primary endpoints in our published experimental settings have weighed in favor of conditioned culture medium, where we have shown that conditioned culture medium has a stimulatory effect on cell survival. In order to explore the reasons for this improved survival we set out to analyze the conditioned culture medium. We utilized ELISA kits to investigate whether brain stem (BS) slice CM contains any significant amounts of brain-derived neurotrophic factor (BDNF) and glial cell derived neurotrophic factor (GDNF). We further looked for a donor cell with progenitor characteristics that would be receptive to BDNF and GDNF. We chose the well-documented boundary cap (BC) progenitor cells to be tested in our in vitro co-culture setting together with cochlear nucleus (CN) of the BS. The results show that BS CM contains BDNF and GDNF and that survival of BC cells, as well as BC cell differentiation into neurons, were enhanced when BS CM were used. Altogether, we conclude that BC cells transplanted into a BDNF and GDNF rich environment could be suitable for treatment of a traumatized or degenerated auditory nerve. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Application of Low cost Spirulina growth medium using Deep sea water

    NASA Astrophysics Data System (ADS)

    Lim, Dae-hack; Kim, Bong-ju; Lee, Sung-jae; Choi, Nag-chul; Park, Cheon-young

    2017-04-01

    Deep-sea water has a relatively constant temperature, abundant nutrients such as calcium, magnesium, nitrates, and phosphates, etc., and stable water quality, even though there might be some variations of their compositions according to collection places. Thus, deep-sea water would be a good substrate for algal growth and biomass production since it contains various nutrients, including a fluorescent red pigment, and β-carotene, etc. The aim of this study was to investigate the economics of a culture condition through comparative analysis to Spirulina platensis growth characteristic under various medium conditions for cost-effective production of Spirulina sp.. Growth experiments were performed with S. platensis under various culture medium conditions (deep sea water + SP medium). Growth tests for culture medium demonstrated that the deep sea water to SP medium ratio of 50:50(W/W) was effective in S. platensis with the maximum biomass (1.35g/L) and minimum medium making cost per production mass (133.28 KRW/g). Parameter estimation of bio-kinetics (maximum growth rate and yield) for low cost medium results showed that the maximum growth rate and yield of N, P, K were obtained under deep sea water to SP medium ratio of 50:50(W/W) of 0.057 1/day and 0.151, 0.076, 0.123, respectively. Acknowledgment : "This research was a part of the project titled 'Development of microalgae culture technique for cosmetic materials based on ocean deep sea water(20160297)', funded by the Ministry of Oceans and Fisheries, Korea."

  5. Demonstration of a Sample Preparation Method for Biological Detection Based on a Novel Membrane Fractionation Technology

    DTIC Science & Technology

    2008-12-31

    TSA 11-g/mL IJ.L IJ.m LWI 2008-01 Acronyms Bacillus globigii also known as Bacillus arophaeus Concentration factor Colony Forming Units per...contracted with MRI to prepare a stock ofBg. This was done using the following steps: 1. An isolation streak was prepared on trypticase soy agar ( TSA ...broth overnight while shaking. 3. Ten milliliters of the overnight culture was inoculated into 1000 mL of a sporulation medium . This sporulation

  6. Pressure determination in Hydrothermal Diamond Anvil Cell via laser interferometry: Investigation of hydrothermal melting of haplogranitic glass

    NASA Astrophysics Data System (ADS)

    Solferino, G.; Anderson, A. J.

    2012-12-01

    Pressure determination in HDAC experiments of hydrothermal melting of a haplogranitic glass at 130-830 MPa and 600-800 °C were performed employing in-situ visualization of alpha to beta quartz via laser interferometry. Hitherto, Raman spectroscopy of ruby, quartz, 13C and zircon has been used for the same purpose, with a best resolution of 40-50 MPa. Our method average uncertainty is just 3.4 MPa. This augmented precision is critical in estimate of the emplacement depth of mid to upper crustal magmatic bodies, e.g., intermediate-felsic intrusions, or definition of formation conditions of magmatic ores, like rare metal pegmatites. Moreover, thanks to this improved resolution on pressure measurements, we observed that actual run pressure, named Pα/β, is smaller than pressure computed using the equation of state (EOS) of pure water, here labeled PH2O for an ample range of pressures, up to 400 MPa. The absolute value of ΔP = Pα/β- PH2O decrease at higher pressure, and switches from negative to positive at P > 800 MPa. Since dissolution of the glass/melt into the pressure medium (water) leads to increment of the medium compressibility (density), then the medium should be able to impose a larger pressure than pure water for every observed temperature of alpha to beta transition (i.e., steeper isochor). A possible explanation of this discrepancy is found in the differential density between the pressure medium and the melt, and in the change of the volume occupied by the fluid for increasing temperature, as it emerges from a simplified model of dissolution of albite feldspar / albite melt in water, prepared for this study on the base of solubility data available in literature.

  7. Detoxification of Hg(II) from aqueous and enzyme media: Pristine vs. tailored calcium alginate hydrogels.

    PubMed

    Sarkar, Kangkana; Ansari, Zarina; Sen, Kamalika

    2016-10-01

    Calcium alginate (CA) hydrogels were tailored using phenolic compounds (PC) like, thymol, morin, catechin, hesperidin, during their preparation. The PC incorporated gels show modified surface features as indicated by scanning electron microscopic images (SEM). The rheological studies show that excepting the hesperidin incorporated gels all the other kinds including calcium alginate pristine have similar mechanical strength. The hesperidine incorporated CA gels had the maximum capacity to adsorb Hg. The Freundlich adsorption isotherms show higher values of adsorption capacity for all PC incorporated CA beads than the pristine CA (PCA). The hesperidin incorporated CA gels were found to show the best adsorption condition at neutral pH and an optimum contact time of 2.5h at 25°C. Considering the possibility of ingested Hg detoxification from human alimentary tract, the hesperidin and morin incorporated CA beads were further modified through incorporation of cod liver oil as the digestion time of fat in stomach is higher. In vitro uptake capacities of Hg in pepsin and pancreatin containing enzyme media were studied with hesperidin and morin incorporated beads and their corresponding fat incorporated beads also. In the pepsin medium, there was no uptake by hesperidin and fat-hesperidin incorporated beads, which is possibly due to the higher acidity of the medium. But in pancreatin medium Hg was taken up by both kinds of beads. Morin and morin-fat incorporated beads were efficient to uptake Hg from both the pepsin and pancreatin medium. The tailored CA beads may therefore serve as efficient scaffolds to rescue Hg ingested individuals. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Enhanced Production of a Recombinant Multidomain Thermostable GH9 Processive Endo-1,4-β-Glucanase (CenC) from Ruminiclostridium thermocellum in a Mesophilic Host Through Various Cultivation and Induction Strategies.

    PubMed

    Haq, Ikram Ul; Akram, Fatima

    2017-09-01

    Commonly, unintentional induction and inadvertently preparing medium for engineered Escherichia coli BL21 CodonPlus (DE3)-RIPL, give poor or variable yields of heterologous proteins. Therefore, to enhance the activity and production of an industrially relevant recombinant processive endo-1,4-β-glucanase (CenC) propagated in Escherichia coli BL21 CodonPlus(DE3)-RIPL through various cultivation and induction strategies. Investigation of various growth media and induction parameters revealed that high-cell-density and optimal CenC expression were obtained in ZYBM9 medium induced either with 0.5 mM IPTG/150 mM lactose, after 6 h induction at 37 °C; and before induction, bacterial cells were given heat shock (42 °C) for 1 h when culture density (OD 600nm ) reached at 0.6. Intracellular enzyme activity was enhanced by 6.67 and 3.20-fold in ZYBM9 and 3×ZYBM9 medium, respectively, under optimal conditions. Using YNG auto-induction medium, activity was 2.5-fold increased after 10 h incubation at 37 °C. Approximately similar results were obtained by transferring the optimized process at the bioreactor level. Results showed that the effective process strategy is essential to enhance recombinant bacterial cell mass and enzyme production from small to large-scale. To the best of our knowledge, this is the first ever report on enhanced production of thermostable processive endo-1,4-β-glucanase cloned from Ruminiclostridium thermocellum, which is a suitable candidate for industrial applications. Graphical Abstract Flow Chart Summary of Enhanced Production of a Recombinant Multidomain Thermostable GH9 Processive Endo-1,4-β-glucanase from Ruminiclostridium thermocellum.

  9. Immobilization of Lipase from Penicillium sp. Section Gracilenta (CBMAI 1583) on Different Hydrophobic Supports: Modulation of Functional Properties.

    PubMed

    Turati, Daniela F M; Morais Júnior, Wilson G; Terrasan, César R F; Moreno-Perez, Sonia; Pessela, Benevides C; Fernandez-Lorente, Gloria; Guisan, Jose M; Carmona, Eleonora C

    2017-02-22

    Lipases are promising enzymes that catalyze the hydrolysis of triacylglycerol ester bonds at the oil/water interface. Apart from allowing biocatalyst reuse, immobilization can also affect enzyme structure consequently influencing its activity, selectivity, and stability. The lipase from Penicillium sp. section Gracilenta (CBMAI 1583) was successfully immobilized on supports bearing butyl, phenyl, octyl, octadecyl, and divinylbenzyl hydrophobic moieties wherein lipases were adsorbed through the highly hydrophobic opened active site. The highest activity in aqueous medium was observed for the enzyme adsorbed on octyl support, with a 150% hyperactivation regarding the soluble enzyme activity, and the highest adsorption strength was verified with the most hydrophobic support (octadecyl Sepabeads), requiring 5% Triton X-100 to desorb the enzyme from the support. Most of the derivatives presented improved properties such as higher stability to pH, temperature, and organic solvents than the covalently immobilized CNBr derivative (prepared under very mild experimental conditions and thus a reference mimicking free-enzyme behavior). A 30.8- and 46.3-fold thermostabilization was achieved in aqueous medium, respectively, by the octyl Sepharose and Toyopearl butyl derivatives at 60 °C, in relation to the CNBr derivative. The octyl- and phenyl-agarose derivatives retained 50% activity after four and seven cycles of p -nitrophenyl palmitate hydrolysis, respectively. Different derivatives exhibited different properties regarding their properties for fish oil hydrolysis in aqueous medium and ethanolysis in anhydrous medium. The most active derivative in ethanolysis of fish oil was the enzyme adsorbed on a surface covered by divinylbenzyl moieties and it was 50-fold more active than the enzyme adsorbed on octadecyl support. Despite having identical mechanisms of immobilization, different hydrophobic supports seem to promote different shapes of the adsorbed open active site of the lipase and hence different functional properties.

  10. [Isolation and characterization of vaginal lactobacilli producing hydrogen peroxide].

    PubMed

    Pashaian, M M; Oganesian, G G

    2011-01-01

    Isolation and characteristics of vaginal lactobacilli that actively generate H2O2 and have high antagonistic activity. Staphylococcus aureus 8956, Escherichia coli 8852, Klebsiella pneumoniae 8795 and Candida albicans 5646 were used as target-strains. Skim milk and MRS medium were used for lactobacilli isolation and cultivation. Antagonism was studied in complete agar and Saburo medium. Merckoquant peroxide test (Merck) stripes were used for the determination of H2O2. Antibacterial activity was determined by diffusion into agar. Specific culture growth rate was determined by conventional method, acidification of the culture medium--by pH-meter. 12 strains were isolated from vaginal smears of healthy women. These strains have an ability to ferment milk among which a highly active H2O2 producer was isolated and attributed to Lactobacillus delbrueckii by the results of 16S rRNA and alpha-subunit RNA polymerase gene sequence analysis (16S rDNA and rpoA genes are registered in GenBank, numbers HQ379171 and HQ379180 respectively). L. delbrueckii MH-10 bacterial cells were characterized by specific growth speed 1.26 per hour, reaching a maximum titer of 2 x 10(9) PFU/ml with lowering medium pH to 4.0. Under aerated conditions H2O2 concentration reached 100 microg/ml or more. L. delbrueckii MH-10 has high antibacterial activity against S. aureus, E. coli, K. pneumoniae. L. delbrueckii MH-10 isolate is an active H2O2 producer, has high growth speed and broad antibacterial activity spectrum, is a perspective candidate for the development of probiotic preparation for the prophylaxis and therapy of vaginoses.

  11. Characterization of an extracellular lipase by Pseudomonas koreensis BK-L07 isolated from soil.

    PubMed

    Anbu, Periasamy

    2014-01-01

    Screening using spirit blue agar revealed that strain BK-L07 had the highest lipase activity. Furthermore, the isolated strain was identified as Pseudomonas sp. based on morphological, physiological, biochemical, and molecular analyses. The 16S rRNA gene sequence of strain BK-L07 shared a high similarity with that of Pseudomonas koreensis (99%). The nutritional conditions and physicochemical properties were influenced by P. koreensis BK-L07. The maximum lipase production was obtained in tryptic soy broth medium at pH 8.0 and a temperature of 25°C after 36 hr of incubation. In addition, the lipase activity was determined using different carbon sources and lipase inducers. The lipase production was greatest when 1% maltose was used as the carbon source and olive oil was used as the lipase inducer. The lipase production was significantly increased approximately threefold in the optimized medium when compared with the original medium. Further, the lipase was purified by ammonium sulfate precipitation and gel filtration chromatography with a purification yield of 10.8%. The molecular mass of lipase was 45 kDa. The optimum temperature and pH were 40°C and 8.0, respectively. The enzyme was stable up to 50°C and at pH from 7 to 9. In addition, the enzyme activity was stimulated by MgSO4 and completely inhibited by ethylenediamine tetraacetic acid (EDTA), indicating the metalloenzyme type. The lipase activity was toward medium to long chain length of fatty acids (C10 to C18). Supplemental materials are available for this article. Go to the publisher's online edition of Preparative Biochemistry and Biotechnology to view the supplemental file.

  12. [Microbial synthesis of deuterium labelled L-phenylalanine with different levels of isotopic enrichment by facultative methylotrophic bacterium Brevibacterium methylicum with RMP assimilation of carbon].

    PubMed

    Mosin, O V; Shvets, V I; Skladnev, D A; Ignatov, I

    2014-01-01

    The preparative microbial synthesis of amino acids labelled with stable isotopes, including deuterium ( 2 H), suitable for biomedical applications by methylotrophic bacteria was studied using L-phenylalanine as example. This amino acid is secreted by Gram-negative aerobic facultative methylotrophic bacteria Brevibacterium methylicum, assimilating methanol via ribulose-5-monophosphate (RMP) cycle of assimilation of carbon, The data on adaptation of L-phenylalanine secreted by methylotrophic bacterium В. methylicum to the maximal concentration of deuterium in the growth medium with 98% 2 Н 2 O and 2% [ 2 Н]methanol, and biosynthesis of deuterium labelled L-phenylalanine With different levels of enrichment are presented. The strain was adapted by means of plating initial cells on firm (2% agarose) minimal growth media with an increasing gradient of 2 Н 2 O concentration from 0; 24.5; 49.0; 73.5 up to 98% 2 Н 2 O followed by subsequent selection of separate colonies stable to the action of 2 Н 2 O. These colonies were capable to produce L-phenylalanine. L-phenylalanine was extracted from growth medium by extraction with isopropanol with the subsequent crystallization in ethanol (output 0.65 g/l). The developed method of microbial synthesis allows to obtain deuterium labelled L-phenylalanine with different levels of isotopic enrichment, depending on concentration of 2 Н 2 O in growth media, from 17% (on growth medium with 24,5% 2 Н 2 O) up to 75% (on growth medium with 98% 2 Н 2 O) of deuterium in the molecule that is confirmed with the data of the electron impact (EI) mass- spectrometry analysis of methyl ethers of N-dimethylamino(naphthalene)-5-sulfochloride (dansyl) phenylalanine in these experimental conditions.

  13. Dimethyl carbonate-mediated lipid extraction and lipase-catalyzed in situ transesterification for simultaneous preparation of fatty acid methyl esters and glycerol carbonate from Chlorella sp. KR-1 biomass.

    PubMed

    Jo, Yoon Ju; Lee, Ok Kyung; Lee, Eun Yeol

    2014-04-01

    Fatty acid methyl esters (FAMEs) and glycerol carbonate were simultaneously prepared from Chlorella sp. KR-1 containing 40.9% (w/w) lipid using a reactive extraction method with dimethyl carbonate (DMC). DMC was used as lipid extraction agent, acyl acceptor for transesterification of the extracted triglycerides, substrate for glycerol carbonate synthesis from glycerol, and reaction medium for the solvent-free reaction system. For 1g of biomass, 367.31 mg of FAMEs and 16.73 mg of glycerol carbonate were obtained under the optimized conditions: DMC to biomass ratio of 10:1 (v/w), water content of 0.5% (v/v), and Novozyme 435 to biomass ratio of 20% (w/w) at 70°C for 24h. The amount of residual glycerol was only in the range of 1-2.5mg. Compared to conventional method, the cost of FAME production with the proposed technique could be reduced by combining lipid extraction with transesterification and omitting the extraction solvent recovery process. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Optimization of culture conditions and medium composition for the production of micrococcin GO5 by Micrococcus sp. GO5.

    PubMed

    Kim, Mi-Hee; Kong, Yoon-Jung; Baek, Hong; Hyun, Hyung-Hwan

    2006-01-02

    To enhance the production of micrococcin GO5, a bacteriocin produced by Micrococcus sp. GO5, cultivation conditions and medium composition were optimized. The optimal initial pH and temperature for bacteriocin production were 7.0-9.0 and 37 degrees C, respectively. Micrococcus sp. GO5 displayed the highest micrococcin GO5 activity when grown in modified MRS medium that contained lactose or sucrose, rather than glucose, as a carbon source. The maximum bacteriocin activity was obtained in modified MRS medium containing 0.5% tryptone and 1.0% yeast extract as nitrogen sources instead of the other nitrogen sources present in MRS medium. Bacteriocin production was greatly affected by the concentration of K(2)HPO(4); strain GO5 produced eight-fold more bacteriocin in medium containing 2.0-2.5% K(2)HPO(4) than in medium containing 0.2% K(2)HPO(4). The optimal concentration of MgSO(4).7H(2)O for bacteriocin production was 0.5%. The production of micrococcin GO5 was increased 32-fold in shake flask culture and 16-fold in a bioreactor using the optimized medium (TY medium), compared with culturing in MRS medium.

  15. Pancreas preservation fluid microbial contamination is associated with poor islet isolation outcomes - a multi-centre cohort study.

    PubMed

    Meier, Raphael P H; Andrey, Diego O; Sun, Pamela; Niclauss, Nadja; Bédat, Benoît; Demuylder-Mischler, Sandrine; Borot, Sophie; Benhamou, Pierre-Yves; Wojtusciszyn, Anne; Buron, Fanny; Pernin, Nadine; Muller, Yannick D; Bosco, Domenico; van Delden, Christian; Berney, Thierry

    2018-03-30

    The microbiological safety of islet preparations is paramount. Preservation medium contamination is frequent, and its impact on islet yield and function remains unclear. Microbiological samples collected during islet isolations from 2006 to 2016 were analyzed and correlated to isolation and allo- and autotransplantation outcomes. Microbial contamination of preservation medium was found in 64.4% of processed donor pancreases (291/452). We identified 464 microorganisms including Staphylococcus (253/464, 54.5%), Streptococcus (31/464, 6.7%), and Candida species (25/464, 5.4%). Microbial contamination was associated with longer warm and cold ischemia times and lower numbers of postpurification islet equivalents, purity, transplant rate, and stimulation index (all P < 0.05). Six percent of the preparations accepted for transplantation showed microbial contamination after isolation (12/200); 9 of 12 were Candida species. Six patients were transplanted with a sample with late microbial growth discovered after the infusion. Insulin independence rate was not affected. This risk of transplanting a contaminated islets preparation was reduced by half following the implementation of an additional sampling after 24 h of islet culture. Pancreas preservation fluid microbial contamination is associated with lower transplant rate and poorer in vitro function, but not with changes in graft survival. Culture medium testing 1 day after isolation reduces the risk of incidental transplantation with contaminated islets. © 2018 Steunstichting ESOT.

  16. The Professional Preparation of Malaysian Teachers in the Implementation of Teaching and Learning of Mathematics and Science in English

    ERIC Educational Resources Information Center

    Idris, Noraini; Cheong, Loh Sau; Nor, Norjoharuddeen Mohd.; Razak, Ahmad Zabidi Abdul; Saad, Rahimi Md.

    2007-01-01

    Malaysia is in the process of changing the medium of instruction for mathematics and science from Malay to English since 2003. To ensure the success of this transition, teachers have to be professional prepared to teach in English. This research aimed to survey the Malaysian science/mathematics teachers' perception towards this professional…

  17. Effects of methadone hydrochloride on the growth of organotypic cerebellar cultures prepared from methadone-tolerant and control rats.

    PubMed

    Willson, N J; Schneider, J F; Roizin, L; Fleiss, J F; Rivers, W; Demartini, J E

    1976-11-01

    Male and female Sprague-Dawley rats were given dl-methadone (5 mg/kg) for at least 3 months and then mated. The drug was continued throughout pregnancy and after delivery. The newly born pups were divided into two groups. One group was tested for in vivo methadone tolerance, while the animals in the othergroup were used to prepare organotypic cerebellar cultures. Various amounts of dl-methadone were added to the media of half of these cerebellum cultures. The effect of the drug in the medium was assessed by measuring explant outgrowth. Similar experiments were carried out with control animals. Statistical analysis of the data obtained in the in vivo portion of the experiment indicates that the pups of methadone-treated mothers tolerate methadone better than those of untreated mothers. The culture experiments revealed that the addition of methadone to the medium reduced explant outgrowth size and this was a dose-related effect. Also, there was significantly less outgrowth from explants prepared using pups of methadone-treated mothers as compared to the controls. There was no significant difference in the effect of methadone on the growth of cultures prepared from the methadone-tolerant and control animals.

  18. A new medium for Caenorhabditis elegans toxicology and nanotoxicology studies designed to better reflect natural soil solution conditions.

    PubMed

    Tyne, William; Lofts, Stephen; Spurgeon, David J; Jurkschat, Kerstin; Svendsen, Claus

    2013-08-01

    A new toxicity test medium for Caenorhabditis elegans is presented. The test solution is designed to provide a better representation of natural soil pore water conditions than currently available test media. The medium has a composition that can readily be modified to allow for studies of the influences of a range of environmentally relevant parameters on nematode biology and toxicology. Tests conducted in the new medium confirmed that nematodes' reproduction was possible at a range of solution pH levels, offering the potential to conduct toxicity studies under a variety of conditions. A test to establish silver nanoparticle and dissolved silver nitrate toxicity, a study type not feasible in M9 or agar media due to precipitation and nanoparticle agglomeration, indicated lower silver nanoparticle (median effective concentration [EC50] of 6.5 mg Ag/L) than silver nitrate (EC50 0.28 mg Ag/L) toxicity. Characterization identified stable nanoparticle behavior in the new test medium. Copyright © 2013 SETAC.

  19. Sensitive determination of 2-methoxyestradiol in pharmaceutical preparations and serum samples using flow injection chemiluminescence.

    PubMed

    Yao, Hanchun; Zhang, Min; Zeng, Wenyuan; Zeng, Xiaoying; Zhang, Zhenzhong

    2014-05-01

    A rapid and sensitive flow injection chemiluminescence (FI-CL) method is described for the determination of 2-methoxyestradiol (2ME) based on enhancement of the CL intensity from a potassium ferricyanide-calcein system in sodium hydroxide medium. The optimum conditions for the CL emission were investigated. Under optimized conditions, a linear calibration graph was obtained over the range 1.0 × 10(-8) to 1.0 × 10(-6) mol/L (r = 0.998) 2ME with a detection limit (3σ) of 5.4 × 10(-9) mol/L. The relative standard deviation (RSD) for 5.0 × 10(-7) mol/L 2ME was 1.7%. As a preliminary application, the proposed method was successfully applied to the determination of 2ME in injection solutions and serum samples. The possible CL mechanism was also proposed. Copyright © 2013 John Wiley & Sons, Ltd.

  20. Analysis of Ricefield Land Damage in Denpasar City, Bali, Indonesia

    NASA Astrophysics Data System (ADS)

    Suyarto, R.; Wiyanti; Dibia, I. N.

    2018-02-01

    Soil as a natural resource, living area, environmental media, and factors of production including biomass production that supports human life and other living beings must be preserved, on the other hand, uncontrolled biomass production activities can cause soil damage, ultimately can threaten the survival of humans and other living things. Therefore, in order to control soil damage, first must inventories the soil condition data and its damage which then visualised in soil damage potential and soil damage status. The activities of the study are the preparation of a map of the initial soil conditions and the delineation of potentially land degradation distribution. Mapping results are used as work maps for verification on the field to take soil samples and create soil damage status. In general, Denpasar City have soil damage potential at very low, low until medium rate. Soil damage status in Denpasar City generally is low damage of bulk volume, total porosity, soil permeability and electrolyte conductivity which beyond limitation thresholds.

  1. Digital-model simulation of the glacial-outwash aquifer, Otter Creek-Dry Creek basin, Cortland County, New York

    USGS Publications Warehouse

    Cosner, O.J.; Harsh, J.F.

    1978-01-01

    The city of Cortland, New York, and surrounding areas obtain water from the highly productive glacial-outwash aquifer underlying the Otter Creek-Dry Creek basin. Pumpage from the aquifer in 1976 was approximately 6.3 million gallons per day and is expected to increase as a result of population growth and urbanization. A digital ground-water model that uses a finite-difference approximation technique to solve partial differential equations of flow through a porous medium was used to simulate the movement of water within the aquifer. The model was calibrated to equilibrium conditions by comparing water levels measured in the aquifer in March 1976 with those computed by the model. Then, from the simulated water-level surface for March, a transient-condition run was made to simulate the surface as measured in September 1976. Computed water levels presented as contours are generally in close agreement with potentiometric-surface maps prepared from field measurements of March and September 1976. (Woodard-USGS)

  2. Oxidative stress differentially impacts male and female bovine embryos depending on the culture medium and the stress condition.

    PubMed

    Dallemagne, Matthew; Ghys, Emmanuelle; De Schrevel, Catalina; Mwema, Ariane; De Troy, Delphine; Rasse, Catherine; Donnay, Isabelle

    2018-09-01

    Male and female embryos are known to differ for their metabolism and response to environmental factors very early in development. The present study aimed to evaluate the response to oxidative stress of male and female bovine embryos at the morula-blastocyst stages in terms of developmental rates, total cell number and apoptotic rates in two culture conditions. Embryos where cultured in a medium supplemented with either 5% fetal calf serum (FCS) or 4 mg/mL bovine serum albumin and a mixture of insulin, transferrin and selenium (BSA-ITS). Oxidative stress was applied at Day-5 post insemination (pi) by adding either AAPH or menadione to the culture medium, and blastocysts were analyzed at Day-7pi. The impact on development and blastocyst quality was dependent on the culture medium and the stress inducer but differed between male and female embryos. Male embryos resisted better to oxidative stress in FCS supplemented medium, no matter the stress inducer. Accordingly, the impact on blastocyst cell number tended to be higher in female blastocysts after stress induction with AAPH in FCS supplemented medium. On the other hand, in BSA-ITS supplemented medium, female embryos were more resistant to AAPH induced stress, while menadione had no impact on sex ratio. The weaker resistance of males to AAPH in this medium is in accordance with their trend to show a higher increase in apoptotic rates than females in this condition. In conclusion, this study shows that oxidative stress has differential impact on male and female bovine blastocysts depending on the culture condition and on the way oxidative stress is induced. Copyright © 2018 Elsevier Inc. All rights reserved.

  3. A Phosphine-mediated Synthesis of 2,3,4,5-tetra-substituted N-hydroxypyrroles from α-oximino Ketones and Dialkyl Acetylenedicarboxylates Under Ionic Liquid Green-media.

    PubMed

    Shahvelayati, Ashraf S; Ghazvini, Maryam; Yadollahzadeh, Khadijeh; Delbari, Akram S

    2018-01-01

    The development of multicomponent reactions (MCRs) in the presence of task-specific ionic liquids (ILs), used not only as environmentally benign reaction media, but also as catalysts, is a new approach that meet with the requirements of sustainable chemistry. In recent years, the use of ionic liquids as a green media for organic synthesis has become a chief study area. This is due to their unique properties such as non-volatility, non-flammability, chemical and thermal stability, immiscibility with both organic compounds and water and recyclability. Ionic liquids are used as environmentally friendly solvents instead of hazardous organic solvents. We report the condensation reaction between α-oximinoketone and dialkyl acetylene dicarboxylate in the presence of triphenylphosphine to afford substituted pyrroles under ionic liquid conditions in good yields. Densely functionalized pyrroles was easily prepared from reaction of α-oximinoketones, dialkyl acetylene dicarboxylate in the presence of triphenylphosphine in a quantitative yield under ionic liquid conditions at room temperature. In conclusion, ionic liquids are indicated as a useful and novel reaction medium for the selective synthesis of functionalized pyrroles. This reaction medium can replace the use of hazardous organic solvents. Easy work-up, synthesis of polyfunctional compounds, decreased reaction time, having easily available-recyclable ionic liquids, and good to high yields are advantages of present method. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  4. ZnO/La 2O 2CO 3 layered composite: A new heterogeneous catalyst for the efficient ultra-fast microwave biofuel production

    DOE PAGES

    Jin, Lei; Zhang, Yashan; Dombrowski, James; ...

    2011-01-21

    The search for solid state materials with high catalytic activities and with no leaching into the reaction medium is one of the key steps toward reducing the cost of producing biodiesel. We report a high biodiesel yield (>95%) in less than 5 min under mild reaction conditions (<100°C) on a ZnO/La₂O₂CO₃ heterogeneous catalyst, showing no catalyst leaching into the reaction medium. The ZnO/La₂O₂CO₃ catalyst is prepared by a co-precipitation method and characterized by X-ray diffraction (XRD), thermogravimetric analyses (TGA), transmission electron microscopy (SEM), and transmission electron microscopy (TEM). The fatty acid methyl ester (FAME) yields as function of different amountsmore » of catalyst was also investigated. Less than 1.0 wt.% catalyst can be used in the reaction to get higher than a 95% FAME yield under mild reaction conditions. The catalytic performance is maintained after storing the catalyst in Ar for a month and no catalyst leaching into the products was found based on XRF analysis. The catalyst has a higher reaction rate than the homogeneous KOH catalyst with the assistance of microwave irradiation. All of these results promote the industrial application of the synthesized ZnO/La₂O₂CO₃ as an ideal catalyst for fast biodiesel production, avoiding many of the issues found in both commercial and independently published catalysts.« less

  5. Development and validation spectroscopic methods for the determination of lomefloxacin in bulk and pharmaceutical formulations

    NASA Astrophysics Data System (ADS)

    El-Didamony, A. M.; Hafeez, S. M.

    2016-01-01

    Four simple, sensitive spectrophotometric and spectrofluorimetric methods (A-D) for the determination of antibacterial drug lomefloxacin (LMFX) in pharmaceutical formulations have been developed. Method A is based on formation of ternary complex between Pd(II), eosin and LMFX in the presence of methyl cellulose as surfactant and acetate-HCl buffer pH 4.0. Spectrophotometrically, under the optimum conditions, the ternary complex showed absorption maximum at 530 nm. Methods B and C are based on redox reaction between LMFX and KMnO4 in acid and alkaline media. In indirect spectrophotometry method B the drug solution is treated with a known excess of KMnO4 in H2SO4 medium and subsequent determination of unreacted oxidant by reacting it with safronine O in the same medium at λmax = 520 nm. Direct spectrophotometry method C involves treating the alkaline solution of LMFX with KMnO4 and measuring the bluish green product at 604 nm. Method D is based on the chelation of LMFX with Zr(IV) to produce fluorescent chelate. At the optimum reaction conditions, the drug-metal chelate showed excitation maxima at 280 nm and emission maxima at 443 nm. The optimum experimental parameters for the reactions have been studied. The validity of the described procedures was assessed. Statistical analysis of the results has been carried out revealing high accuracy and good precision. The proposed methods were successfully applied for the determination of the selected drug in pharmaceutical preparations with good recoveries.

  6. Microgravity and bone cell mechanosensitivity: FLOW experiment during the DELTA mission

    NASA Astrophysics Data System (ADS)

    Bacabac, Rommel G.; Van Loon, Jack J. W. A.; de Blieck-Hogervorst, Jolanda M. A.; Semeins, Cor M.; Zandieh-Doulabi, Behrouz; Helder, Marco N.; Smit, Theo H.; Klein-Nulend, Jenneke

    2007-09-01

    The catabolic effects of microgravity on mineral metabolism in bone organ cultures might be explained as resulting from an exceptional form of disuse. It is possible that the mechanosensitivity of bone cells is altered under near weightlessness conditions, which likely contributes to disturbed bone metabolism observed in astronauts. In the experiment "FLOW", we tested whether the production of early signaling molecules that are involved in the mechanical load-induced osteogenic response by bone cells is changed under microgravity conditions. FLOW was one of the Biological experiment entries to the Dutch Soyuz Mission "DELTA" (Dutch Expedition for Life Science, Technology and Atmospheric Research). FLOW was flown by the Soyuz craft, launched on April 19, 2004, on its way to the International Space Station. Primary osteocytes, osteoblasts, and periosteal fibroblasts were incubated in plunger boxes, developed by Centre for Concepts in Mechatronics, using plunger activation events for single pulse fluid shear stress stimulations. Due to unforeseen hardware complications, results from in-flight cultures are considered lost. Ground control experiments showed an accumulative increase of NO in medium for osteocytes (as well as for osteoblasts and periosteal fibroblasts). Data from the online-NO sensor showed that the NO produced in medium by osteocytes increased sharply after pulse shear stress stimulations. COX-2 mRNA expression revealed high levels in osteoblasts compared to the other cell types tested. In conclusion, preparations for the FLOW experiment and preliminary ground results indicate that the FLOW setup is viable for a future flight opportunity.

  7. A study on the effects of temperature and volume fraction on thermal conductivity of copper oxide nanofluid.

    PubMed

    Jwo, Ching-Song; Chang, Ho; Teng, Tun-Ping; Kao, Mu-Jnug; Guo, Yu-Ting

    2007-06-01

    By using copper oxide nanofluid fabricated by the self-made Submerged Arc Nanofluid Synthesis System (SANSS), this paper measures the thermal conductivity under different volume fractions and different temperatures by thermal properties analyzer, and analyzes the correlation among the thermal conductivity, volume fraction, and temperature of nanofluid. The CuO nanoparticles used in the experiment are needle-like, with a mean particle size of about 30 nm. They can be stably suspended in deionized water for a long time. The experimental results show that under the condition that the temperature is 40 degrees C, when the volume fraction of nanofluid increases from 0.2% to 0.8%, the thermal conductivity increment of the prepared nanofluid towards deionized water can be increased from 14.7% to 38.2%. Under the condition that the volume fraction is 0.8%, as the temperature of nanofluid rises from 5 degrees C to 40 degrees C, the thermal conductivity increment of the prepared nanofluid towards deionized water increases from 5.9% to 38.2%. Besides, the effects of temperature change are greater than the effects of volume fraction on the thermal conductivity of nanofluid. Therefore, when the self-made copper oxide nanofluid is applied to the heat exchange device under medium and high temperature, an optimal radiation effect can be acquired.

  8. Tunable pH and redox-responsive drug release from curcumin conjugated γ-polyglutamic acid nanoparticles in cancer microenvironment.

    PubMed

    Pillarisetti, Shameer; Maya, S; Sathianarayanan, S; Jayakumar, R

    2017-11-01

    Tunable pH and redox responsive polymer was prepared using γ-polyglutamic acid (γ-PGA) with linker 3-mercaptopropionic acid (3-MPA) (γ-PGA_SH) via oxidation to obtain redox responsive disulfide (γ-PGA_SS) backbone and adipic acid dihydrazide (ADH) (γ-PGA_SS_ADH) with hydrazide functional group for pH responsiveness. Further curcumin (Cur) was conjugated through hydrazone bond of the γ-PGA_SS_ADH via Schiff base reaction to obtain (γ-PGA_SS_ADH_Cur). The prepared systems were characterized by Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, Electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Qq-TOF-MS/MS) and Solid state nuclear magnetic resonance (SS NMR) techniques. γ-PGA_SS_ADH_Cur formed self-assembled core shell nanoparticles (NPs) in existence of stabilized aqueous medium. γ-PGA_SS_ADH_Cur NPs maintained its stability in physiological condition. NPs tunable Cur release and cytotoxicity were observed for γ-PGA_SS_ADH_Cur NPs in both acidic and redox conditions mimicking the cancer microenvironment. γ-PGA_SS_ADH_Cur NPs uptake study showed via endocytosis mechanism resulted in the lysosomal entrapment of these NPs within the cell. γ-PGA_SS_ADH_Cur NPs exhibited a dual stimuli responsive drug delivery and can be used as a smart and potential drug delivery system in cancer microenvironment. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Characterization of recombinant prolyl aminopeptidase from Aspergillus oryzae.

    PubMed

    Matsushita-Morita, M; Furukawa, I; Suzuki, S; Yamagata, Y; Koide, Y; Ishida, H; Takeuchi, M; Kashiwagi, Y; Kusumoto, K-I

    2010-07-01

    Prolyl aminopeptidase (PAP) degrades only amino-terminal proline from peptides. The food-grade fungus Aspergillus oryzae produces this enzyme only in small amounts. In this paper, we present efficient production of recombinant PAP with an overexpression system of A. oryzae and characterization of its biochemical properties. The gene encoding PAP was overexpressed as a His-tag fusion protein under a taka-amylase gene (amyB) promoter with a limited expressing condition in A. oryzae. The PAP activity in the mycelia grown in rich medium containing glucose (repressing condition) was twice that in starch (inducing condition). The enzyme prepared as cell-free extract was partially purified through two-step column chromatography. The PAP was estimated to be a hexameric protein and exhibited salt tolerance against NaCl of up to 4 mol l(-1). Aspergillus oryzae PAP was produced under the repressing condition of amyB promoter in a PAP-overexpressing strain and purified 1800-folds. Overproduction of PAP under promoter-inducing conditions led to an increase in inactive PAP, possibly because of irregular folding. PAP with a high specific activity and salt tolerance may be used effectively in the manufacturing processes of fermented foods. Journal compilation © 2009 The Society for Applied Microbiology. No claim to Japanese Government works.

  10. Lemna paucicostata Hegelm. 6746

    PubMed Central

    Datko, Anne H.; Mudd, S. Harvey; Giovanelli, John

    1980-01-01

    Photoautotrophic and mixotrophic growth of Lemna paucicostata Hegelm. 6746 (formerly Lemna perpusilla Torr. 6746) was investigated to establish standardized conditions for biochemical studies. Optimal temperature for growth was 29 to 30 C. The medium used previously (Datko AH, Mudd SH, Giovanelli J 1977 J Biol Chem 252: 3436-3445) was modified by inclusion of NH4Cl, decreasing macronutrient and ethylenediamine tetraacetate concentration, increasing micronutrient concentration, and inclusion of bicarbonate (for photoautotrophic growth) or 2-(N-morpholino)ethanesulfonic acid (for mixotrophic growth) buffers. Varying the sulfate concentration between 14 and 1 millimolar had no effect on growth. For photoautotrophic growth in the new medium (medium 4), the effects of CO2 concentration, light intensity, and pH were measured. Under the optimal conditions, a multiplication rate (MR) of 300 to 315, equivalent to a doubling time of 23 to 24 hours was obtained. Addition of glutamine or asparagine did not increase this MR. For mixotrophic growth in low light, the effects of sucrose concentration and pH were determined. Under optimal conditions, MR was 210. A concentration of sucrose less than maximal for growth was chosen for the medium for experiments which will include 14C-labeling of intermediates. MR under these conditions was 184. Growth was equally good in medium 4 and in half-strength Hutner's medium when sulfate was high (0.4 to 1 millimolar), but better in medium 4 when sulfate was low (20 micromolar). Growth rates could be restored to normal in half-strength Hutner's with low sulfate by decreasing the molybdate concentration. By modifying medium 4 to contain very low amounts of sulfate, and by preconditioning medium and plants, it was shown that there was an increment in plant protein of approximately 2.5 micrograms per nanomole of added MgSO4. Colonies undergoing sulfur limitation exhibited a slow growth rate and a high frond to colony ratio. Molybdate and selenate produced growth inhibition reversible by sulfate. Conditions were developed in which the plants could be maintained indefinitely in the presence of either molybdate or selenate in altered metabolic steady-states with lowered growth rates and protein per frond. Images PMID:16661306

  11. Laboratory tools and e-learning elements in training of acousto-optics

    NASA Astrophysics Data System (ADS)

    Barócsi, Attila; Lenk, Sándor; Ujhelyi, Ferenc; Majoros, Tamás.; Maák, Paál.

    2015-10-01

    Due to the acousto-optic (AO) effect, the refractive index of an optical interaction medium is perturbed by an acoustic wave induced in the medium that builds up a phase grating that will diffract the incident light beam if the condition of constructive interference is satisfied. All parameters, such as magnitude, period or phase of the grating can be controlled that allows the construction of useful devices (modulators, switches, one or multi-dimensional deflectors, spectrum analyzers, tunable filters, frequency shifters, etc.) The research and training of acousto-optics have a long-term tradition at our department. In this presentation, we introduce the related laboratory exercises fitted into an e-learning frame. The BSc level exercise utilizes a laser source and an AO cell to demonstrate the effect and principal AO functions explaining signal processing terms such as amplitude or frequency modulation, modulation depth and Fourier transformation ending up in building a free space sound transmitting and demodulation system. The setup for MSc level utilizes an AO filter with mono- and polychromatic light sources to learn about spectral analysis and synthesis. Smart phones can be used to generate signal inputs or outputs for both setups as well as to help students' preparation and reporting.

  12. Functionalized Agarose Self-Healing Ionogels Suitable for Supercapacitors.

    PubMed

    Trivedi, Tushar J; Bhattacharjya, Dhrubajyoti; Yu, Jong-Sung; Kumar, Arvind

    2015-10-12

    Agarose has been functionalized (acetylated/carbanilated) in an ionic liquid (IL) medium of 1-butyl-3-methylimidazolium acetate at ambient conditions. The acetylated agarose showed a highly hydrophobic nature, whereas the carbanilated agarose could be dissolved in water as well as in the IL medium. Thermoreversible ionogels were obtained by cooling the IL sols of carbanilated agarose at room temperature. The ionogel prepared from a protic-aprotic mixed-IL system (1-butyl-3-methylimidazolium chloride and N-(2-hydroxyethyl)ammonium formate) demonstrated a superior self-healing property, as confirmed from rheological measurements. The superior self-healing property of such an ionogel has been attributed to the unique inter-intra hydrogen-bonding network of functional groups inserted in the agarose. The ionogel was tested as a flexible solid electrolyte for an activated-carbon-based supercapacitor cell. The measured specific capacitance was found to be comparable with that of a liquid electrolyte system at room temperature and was maintained for up to 1000 charge-discharge cycles. Such novel functionalized-biopolymer self-healing ionogels with flexibility and good conductivity are desirable for energy-storage devices and electronic skins with superior lifespans and robustness. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. [Determination of peracetic acid and hydrogen peroxide in a preparation].

    PubMed

    Bodiroga, Milanka; Ognjanović, Jasminka

    2002-01-01

    Iodometric and permanganometric titrations were used for determination of peracetic acid and hydrogen peroxide (H2O2) in the mixture. Two procedures were described and compared. Titrations could be done in only one vessel, in the same reaction mixture, when iodometric titration of peracetic acid was continued after the permanganometric titration of H2O2, (procedure A). Peracetic acid and H2O2, as oxidizing agents, reacted with potassium iodide in an acid medium, evolving iodine. This reaction was used for the quantitative iodometric determination of total peroxide in procedure B. H2O2 reacted with potassium permanganate in acid medium, but peracetic acid did not react under the same conditions. That made possible the selective permanganometric determination of H2O2 in the presence of peracetic acid. The procedure B was performed in two titration vessels (KV = 3.4% for peracetic acid, 0.6% for H2O2). The procedure A for iodometric determination of peracetic acid in one titration vessel after permanganometric titration of H2O2 was recommended (KV = 2.5% for peracetic acid, 0.45% for H2O2).

  14. [Effects of environmental stress on secondary metabolites of Aspergillus ochraceus LCJ11-102 associated with the coral Dichotella gemmacea].

    PubMed

    Zhou, Yalin; Wang, Yi; Liu, Peipei; Wang, Zhiying; Zhu, Weiming

    2010-08-01

    To explore the secondary metabolites of fungus Aspergillus ochraceus LCJ11-102 associated with the coral Dichotella gemmacea under environmental stress and to obtain characteristic compounds with biological activities. A nutrient-deprived culture medium (biomimetic culture) and a high salt culture medium were used for fermentation. Fingerprints of HPLC of the fermentation broth were used to investigate the diversity of secondary metabolites. Compounds were isolated by column chromatography on silica gel, Sephadex LH-20, and preparative HPLC. Their structures were identified by spectroscopic analyses and the modified Mosher's method. Different secondary metabolites were produced by A. ochraceus LCJ11-102 under two different culture conditions. (R)-mellein (1), (5,6-trans, 8,9-threo-) -9-chloro-8-hydroxy-8, 9-deoxyaspyrone (2), (5,6-erythro-, 8,9-threo-) -9-chloro-8-hydroxy-8, 9-deoxyasperlactone (3), and (5S, 6R, 9S)-dihydroaspyrone (4) were identified from the biomimetic cultures, and R (+) -semi-vioxanthin (5) was identified from the high salt cultures, respectively. Environmental stress obviously induces microbes to produce different secondary metabolites. And biomimetic culture is an effective approach to obtain active chloro compounds from marine microorganisms.

  15. Simple method for culture of peripheral blood lymphocytes of Testudinidae.

    PubMed

    Silva, T L; Silva, M I A; Venancio, L P R; Zago, C E S; Moscheta, V A G; Lima, A V B; Vizotto, L D; Santos, J R; Bonini-Domingos, C R; Azeredo-Oliveira, M T V

    2011-12-06

    We developed and optimized a simple, efficient and inexpensive method for in vitro culture of peripheral blood lymphocytes from the Brazilian tortoise Chelonoidis carbonaria (Testudinidae), testing various parameters, including culture medium, mitogen concentration, mitotic index, culture volume, incubation time, and mitotic arrest. Peripheral blood samples were obtained from the costal vein of four couples. The conditions that gave a good mitotic index were lymphocytes cultured at 37°C in minimum essential medium (7.5 mL), with phytohemagglutinin as a mitogen (0.375 mL), plus streptomycin/penicillin (0.1 mL), and an incubation period of 72 h. Mitotic arrest was induced by 2-h exposure to colchicine (0.1 mL), 70 h after establishing the culture. After mitotic arrest, the cells were hypotonized with 0.075 M KCl for 2 h and fixed with methanol/acetic acid (3:1). The non-banded mitotic chromosomes were visualized by Giemsa staining. The diploid chromosome number of C. carbonaria was found to be 52 in females and males, and sex chromosomes were not observed. We were able to culture peripheral blood lymphocytes of a Brazilian tortoise in vitro, for the preparation of mitotic chromosomes.

  16. Fast nucleation for silica nanoparticle synthesis using a sol-gel method.

    PubMed

    Dixit, Chandra K; Bhakta, Snehasis; Kumar, Ajeet; Suib, Steven L; Rusling, James F

    2016-12-01

    We have developed a method that for the first time allowed us to synthesize silica particles in 20 minutes using a sol-gel preparation. Therefore, it is critically important to understand the synthesis mechanism and kinetic behavior in order to achieve a higher degree of fine tuning ability during the synthesis. In this study, we have employed our ability to modulate the physical nature of the reaction medium from sol-gel to emulsion, which has allowed us to halt the reaction at a particular time; this has allowed us to precisely understand the mechanism and chemistry of the silica polymerization. The synthesis medium is kept quite simple with tetraethyl orthosilicate (TEOS) as a precursor in an equi-volumetric ethanol-water system and with sodium hydroxide as a catalyst. Synthesis is performed under ambient conditions at 20 °C for 20 minutes followed by phasing out of any unreacted TEOS and polysilicic acid chains via their emulsification with supersaturated water. We have also demonstrated that the developed particles with various sizes can be used as seeds for further particle growth and other applications. Luminol, a chemiluminescent molecule, has been entrapped successfully between the layers of silica and was demonstrated for the chemiluminescence of these particles.

  17. The effect of environmental and physiological conditions on excystation of Acanthamoeba castellanii belonging to the T4 genotype.

    PubMed

    Lakhundi, Sahreena; Khan, Naveed Ahmed; Siddiqui, Ruqaiyyah

    2014-08-01

    Excystation in Acanthamoeba is an important property for the onset of infection as well as infection recurrence, post-treatment. The overall aim of this study was to determine the effects of several environmental and physiological parameters on excystation in Acanthamoeba castellanii belonging to the T4 genotype. Cysts were prepared by inoculating A. castellanii trophozoites on non-nutrient agar plates for up to 2 weeks. To determine the effects of various conditions on excystation, A. castellanii cysts were inoculated in growth medium i.e. PYG and incubated at varying temperatures (4-40 °C), various pHs (4-9), artificial light/dark cycles and 5% of CO2. Optimum excystation was observed when cysts were incubated at 30 °C in growth medium at neutral pH. Extremes of temperature and pH reduced excystation, while light/dark cycles had no effect on excystation of A. castellanii. On the other hand, 5% of CO2 enhanced excystation and growth of excysting amoebae. To determine the effect of serum on A. castellanii excystation, assays were performed in the presence of varying concentrations of heat-inactivated foetal bovine serum (FBS) (5-100%). The results revealed that FBS promoted excystation. The involvement of G proteins in excystation was also determined. Using propranolol hydrochloride, a G protein inhibitor, the results revealed that G proteins play a role in A. castellanii differentiation. Furthermore, organic solvents (methanol/ethanol) completely blocked excystation. None of the aforementioned conditions had any effect on the viability of A. castellanii. A complete understanding of excystation in A. castellanii will be of value to counter infection recurrence.

  18. Corrosion of 2205 Duplex Stainless Steel Weldment in Chloride Medium Containing Sulfate-Reducing Bacteria

    NASA Astrophysics Data System (ADS)

    Antony, P. J.; Singh Raman, R. K.; Kumar, Pradeep; Raman, R.

    2008-11-01

    Influence of changes in microstructure caused due to welding on microbiologically influenced corrosion of a duplex stainless steel was studied by exposing the weldment and parent metal to chloride medium containing sulfate-reducing bacteria (SRB). Identically prepared coupons (same area and surface finish) exposed to sterile medium were used as the control. Etching-type attack was observed in the presence of SRB, which was predominant in the heat-affected zone (HAZ) of the weldment. The anodic polarization studies indicated an increase in current density for coupon exposed to SRB-containing medium as compared to that obtained for coupon exposed to sterile medium. The scanning electron microscopy (SEM) observations after anodic polarization revealed that the attack was preferentially in the ferrite phase of HAZ of the weldment, whereas it was restricted to the austenite phase of the parent metal.

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

    Fox, Sandra Lynn; Bala, Greg Alan

    Surfactin, a lipopeptide biosurfactant, produced by Bacillus subtilis is known to reduce the surface tension of water from 72 to 27 mN/m. Potato substrates were evaluated as a carbon source for surfactant production by B. subtilis ATCC 21332. An established potato medium, simulated liquid and solid potato waste media, and a commercially prepared potato starch in a mineral salts medium were evaluated in shake flask experiments to verify growth, surface tension reduction, and carbohydrate reduction capabilities. Total carbohydrate assays and glucose monitoring indicated that B. subtilis was able to degrade potato substrates to produce surfactant. Surface tensions dropped from 71.3±0.1more » to 28.3±0.3 mN/m (simulated solid potato medium) and to 27.5±0.3 mN/m (mineral salts medium). A critical micelle concentration (CMC) of 0.10 g/l was obtained from a methylene chloride extract of the simulated solid potato medium.« less

  20. Fabrication of hydrogels with elasticity changed by alkaline phosphatase for stem cell culture.

    PubMed

    Toda, Hiroyuki; Yamamoto, Masaya; Uyama, Hiroshi; Tabata, Yasuhiko

    2016-01-01

    The objective of this study is to design hydrogels whose elasticity can be changed by alkaline phosphatase (ALP) in cell culture and evaluate the effect of hydrogel elasticity on an osteogenic gene expression of cells. Hydrogels were prepared by the radical polymerization of acrylamide (AAm), N,N'-methylenebisacrylamide (BIS), and Phosmer™M containing phosphate groups (PE-PAAm hydrogels). The storage modulus of PE-PAAm hydrogels prepared was changed by the preparation conditions. When human mesenchymal stem cells (hMSC) were cultured on the ALP-responsive PE-PAAm hydrogels in the presence or absence of ALP, the morphology of hMSC was observed and one of the osteogenic differentiation markers, Runx2, was evaluated. By ALP addition into the culture medium, the morphology of hMSC was changed into an elongated shape without cell damage. ALP addition modified the level of Runx2 gene expression, which was influenced by the modulus of PE-PAAm hydrogels. It is concluded that the elasticity change of hydrogel substrates in cell culture had an influence on the Runx2 gene expression of hMSC. Stem cells sense the surface elasticity of culture substrates, and their differentiation fate is biologically modified by substrate properties. Most of experiments have been performed in static conditions during cell culture, while the in vivo microenvironment is dynamically changed. In this study, we established to design an enzyme-responsive hydrogel whose elasticity can be changed by alkaline phosphatase (ALP) in cell culture to mimic in vivo conditions. As a result, the cells were deformed and the gene expression level of an osteogenic maker, Runx2, was modified by ALP treatment. This is the novel report describing to demonstrate that the dynamic alteration of hydrogel substrate elasticity could modulate the osteoblastic gene expression of human MSC in vitro. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  1. Stimulation of GABA-Induced Ca2+ Influx Enhances Maturation of Human Induced Pluripotent Stem Cell-Derived Neurons

    PubMed Central

    Rushton, David J.; Mattis, Virginia B.; Svendsen, Clive N.; Allen, Nicholas D.; Kemp, Paul J.

    2013-01-01

    Optimal use of patient-derived, induced pluripotent stem cells for modeling neuronal diseases is crucially dependent upon the proper physiological maturation of derived neurons. As a strategy to develop defined differentiation protocols that optimize electrophysiological function, we investigated the role of Ca2+ channel regulation by astrocyte conditioned medium in neuronal maturation, using whole-cell patch clamp and Ca2+ imaging. Standard control medium supported basic differentiation of induced pluripotent stem cell-derived neurons, as assayed by the ability to fire simple, single, induced action potentials. In contrast, treatment with astrocyte conditioned medium elicited complex and spontaneous neuronal activity, often with rhythmic and biphasic characteristics. Such augmented spontaneous activity correlated with astrocyte conditioned medium-evoked hyperpolarization and was dependent upon regulated function of L-, N- and R-type Ca2+ channels. The requirement for astrocyte conditioned medium could be substituted by simply supplementing control differentiation medium with high Ca2+ or γ-amino butyric acid (GABA). Importantly, even in the absence of GABA signalling, opening Ca2+ channels directly using Bay K8644 was able to hyperpolarise neurons and enhance excitability, producing fully functional neurons. These data provide mechanistic insight into how secreted astrocyte factors control differentiation and, importantly, suggest that pharmacological modulation of Ca2+ channel function leads to the development of a defined protocol for improved maturation of induced pluripotent stem cell-derived neurons. PMID:24278369

  2. Efficient Preparation of Streptochlorin from Marine Streptomyces sp. SYYLWHS-1-4 by Combination of Response Surface Methodology and High-Speed Counter-Current Chromatography.

    PubMed

    Li, Lin; He, Shan; Ding, Lijian; Yuan, Ye; Zhu, Peng; Epstein, Slava; Fan, Jianzhong; Wu, Xiaokai; Yan, Xiaojun

    2016-05-27

    Since first isolated from the lipophilic extract of Streptomyces sp. SF2583, streptochlorin, has attracted a lot of attention because of its various pharmacological properties, such as antibiotic, antiallergic, antitumor, and anti-inflammatory activities. For the efficient preparation of streptochlorin from a producing strain Streptomyces sp. SYYLWHS-1-4, we developed a combinative method by using response surface methodology (RSM) and high-speed counter-current chromatography (HSCCC). In the fermentation process, we used RSM to optimize the condition for the efficient accumulation of streptochlorin, and the optimal parameters were: yeast extract 1.889 g/L, soluble starch 8.636 g/L, K₂HPO₄ 0.359 g/L, CaCl₂ 2.5 g/L, MgSO₄ 0.625 g/L, marine salt 25 g/L, medium volume 50%, initial pH value 7.0, temperature 27.5 °C, which enhanced streptochlorin yield by 17.7-fold. During the purification process, the preparative HSCCC separation was performed using a petroleum ether-ethyl acetate-methanol-water (9:0.8:5:5, v/v/v/v) biphasic solvent system, where 300 mg of crude sample yielded 16.5 mg streptochlorin with over 95% purity as determined by UPLC. Consequently, the combination method provided a feasible strategy for highly effective preparation of streptochlorin, which ensured the supply of large amounts of streptochlorin for in vivo pharmacological assessments or other requirements.

  3. The in vitro isolated whole guinea pig brain as a model to study epileptiform activity patterns.

    PubMed

    de Curtis, Marco; Librizzi, Laura; Uva, Laura

    2016-02-15

    Research on ictogenesis is based on the study of activity between seizures and during seizures in animal models of epilepsy (chronic condition) or in in vitro slices obtained from naïve non-epileptic brains after treatment with pro-convulsive drugs, manipulations of the extracellular medium and specific stimulation protocols. The in vitro isolated guinea pig brain retains the functional connectivity between brain structures and maintains interactions between neuronal, glial and vascular compartments. It is a close-to-in vivo preparation that offers experimental advantages not achieved with the use of other experimental models. Neurophysiological and imaging techniques can be utilized in this preparation to study brain activity during and between seizures induced by pharmacological or functional manipulations. Cellular and network determinants of interictal and ictal discharges that reproduce abnormal patterns observed in human focal epilepsies and the associated changes in extracellular ion and blood-brain permeability can be identified and analyzed in the isolated guinea pig brain. Ictal and interictal patterns recorded in in vitro slices may show substantial differences from seizure activity recorded in vivo due to slicing procedure itself. The isolated guinea pig brain maintained in vitro by arterial perfusion combines the typical facilitated access of in vitro preparations, that are difficult to approach during in vivo experiments, with the preservation of larger neuronal networks. The in vitro whole isolated guinea pig brain preparation offers an unique experimental model to study systemic and neurovascular changes during ictogenesis. Published by Elsevier B.V.

  4. Measurements of bovine sperm velocities under true anaerobic and aerobic conditions.

    PubMed

    Krzyzosiak, J; Molan, P; Vishwanath, R

    1999-04-30

    Velocities of bovine spermatozoa in a medium containing glucose were similar under true anaerobic and aerobic conditions. Spermatozoa were not able to sustain motility under anaerobic conditions when glycolysis was inhibited, but regained motility when re-aerated. This demonstrates that immobilisation was due to lack of oxygen and that conditions under which motility was analysed were truly anaerobic. Sperm motility parameters were not significantly different in the presence and absence of 4 microM antimycin A and 4 microM rotenone when glucose was present in the medium. After each incubation, functionality of sperm mitochondria was assayed by washing sperm into the medium which supported respiration but not glycolysis, and motility was visually assessed. All sperm samples were highly motile in this medium indicating that their mitochondria were functional. When glycolysis was inhibited, antimycin and rotenone abolished sperm motility immediately after addition. Bovine sperm can maintain similar levels of motility aerobically and anaerobically if a glycolysable substrate is available. Available data on bovine sperm energetics support this view.

  5. A simplified radiometabolite analysis procedure for PET radioligands using a solid phase extraction with micellar medium.

    PubMed

    Nakao, Ryuji; Halldin, Christer

    2013-07-01

    A solid phase extraction method has been developed for simple and high-speed direct determination of PET radioligands in plasma. This methodology makes use of a micellar medium and a solid-phase extraction cartridge for displacement of plasma protein bound radioligand and separation of PET radioligands from their radiometabolites without significant preparation. The plasma samples taken from monkey or human during PET measurements were mixed with a micellar eluent containing an anionic surfactant sodium dodecyl sulphate and loaded onto SPE cartridges. The amount of radioactivity corresponding to parent radioligand (retained on the cartridge) and its radioactive metabolites (eluted with micellar eluent) was measured. Under the optimized conditions, excellent separation of target PET radioligands from their radiometabolites was achieved with a single elution and short run-time of 1 min. This method was successfully applied to study the metabolism for (11)C-labelled radioligands in human or monkey plasma. The amount of parent PET radioligands estimated by micellar solid phase extraction strongly corresponded with that determined by radio-LC. The improved throughput permitted the analysis of a large number of plasma samples (up to 13 samples per one PET study) for accurate estimation of metabolite-corrected input function during quantitative PET imaging studies. Solid phase extraction together with micellar medium is fast, sensitive and easy to use, and therefore it is an attractive alternative method to determine relative composition of PET radioligands in plasma. Copyright © 2013 Elsevier Inc. All rights reserved.

  6. Modulation of antimicrobial metabolites production by the fungus Aspergillus parasiticus

    PubMed Central

    Bracarense, Adriana A.P.; Takahashi, Jacqueline A.

    2014-01-01

    Biosynthesis of active secondary metabolites by fungi occurs as a specific response to the different growing environments. Changes in this environment alter the chemical and biological profiles leading to metabolites diversification and consequently to novel pharmacological applications. In this work, it was studied the influence of three parameters (fermentation length, medium composition and aeration) in the biosyntheses of antimicrobial metabolites by the fungus Aspergillus parasiticus in 10 distinct fermentation periods. Metabolism modulation in two culturing media, CYA and YES was evaluated by a 22 full factorial planning (ANOVA) and on a 23 factorial planning, role of aeration, medium composition and carbohydrate concentration were also evaluated. In overall, 120 different extracts were prepared, their HPLC profiles were obtained and the antimicrobial activity against A. flavus, C. albicans, E. coli and S. aureus of all extracts was evaluated by microdilution bioassay. Yield of kojic acid, a fine chemical produced by the fungus A. parasiticus was determined in all extracts. Statistical analyses pointed thirteen conditions able to modulate the production of bioactive metabolites by A. parasiticus. Effect of carbon source in metabolites diversification was significant as shown by the changes in the HPLC profiles of the extracts. Most of the extracts presented inhibition rates higher than that of kojic acid as for the extract obtained after 6 days of fermentation in YES medium under stirring. Kojic acid was not the only metabolite responsible for the activity since some highly active extracts showed to possess low amounts of this compound, as determined by HPLC. PMID:24948950

  7. Pharmacological profiling of the TRPV3 channel in recombinant and native assays.

    PubMed

    Grubisha, Olivera; Mogg, Adrian J; Sorge, Jessica L; Ball, Laura-Jayne; Sanger, Helen; Ruble, Cara L A; Folly, Elizabeth A; Ursu, Daniel; Broad, Lisa M

    2014-05-01

    Transient receptor potential vanilloid subtype 3 (TRPV3) is implicated in nociception and certain skin conditions. As such, it is an attractive target for pharmaceutical research. Understanding of endogenous TRPV3 function and pharmacology remains elusive as selective compounds and native preparations utilizing higher throughput methodologies are lacking. In this study, we developed medium-throughput recombinant and native cellular assays to assess the detailed pharmacological profile of human, rat and mouse TRPV3 channels. Medium-throughput cellular assays were developed using a Ca(2+) -sensitive dye and a fluorescent imaging plate reader. Human and rat TRPV3 pharmacology was examined in recombinant cell lines, while the mouse 308 keratinocyte cell line was used to assess endogenous TRPV3 activity. A recombinant rat TRPV3 cellular assay was successfully developed after solving a discrepancy in the published rat TRPV3 protein sequence. A medium-throughput, native, mouse TRPV3 keratinocyte assay was also developed and confirmed using genetic approaches. Whereas the recombinant human and rat TRPV3 assays exhibited similar agonist and antagonist profiles, the native mouse assay showed important differences, namely, TRPV3 activity was detected only in the presence of potentiator or during agonist synergy. Furthermore, the native assay was more sensitive to block by some antagonists. Our findings demonstrate similarities but also notable differences in TRPV3 pharmacology between recombinant and native systems. These findings offer insights into TRPV3 function and these assays should aid further research towards developing TRPV3 therapies. © 2013 The British Pharmacological Society.

  8. Germ tube and chlamydospore formation by Candida albicans on a new medium.

    PubMed Central

    Beheshti, F; Smith, A G; Krause, G W

    1975-01-01

    A new medium composed of "cream of rice" infusion, oxgall, Tween 80, and agar is described for the sequential development of germ tubes and chlamydospores by Candida albicans. The procedure used (Dalmau's technique) is an improvement over the fluid substrate procedures previously advocated for germ tube formation. That the same preparation is then used for chlamydospore production is of practical importance for the clinical mycology laboratory. Images PMID:1102561

  9. Liquid chromatographic extraction medium

    DOEpatents

    Horwitz, E.P.; Dietz, M.L.

    1994-09-13

    A method and apparatus are disclosed for extracting strontium and technetium values from biological, industrial and environmental sample solutions using a chromatographic column. An extractant medium for the column is prepared by generating a solution of a diluent containing a Crown ether and dispersing the solution on a resin substrate material. The sample solution is highly acidic and is introduced directed to the chromatographic column and strontium or technetium is eluted using deionized water. 1 fig.

  10. Optimization of Carbon and Nitrogen Sources for Extracellular Polymeric Substances Production by Chryseobacterium indologenes MUT.2

    PubMed Central

    Khani, Mojtaba; Bahrami, Ali; Chegeni, Asma; Ghafari, Mohammad Davoud; Mansouran Zadeh, ALi

    2016-01-01

    Background Bacterial Extracellular Polymeric Substances (EPS) are environmental friendly and versatile polymeric materials that are used in a wide range of industries such as: food, textile, cosmetics, and pharmaceuticals. To make the production process of the EPS cost-effective, improvements in the production yield is required which could be implemented through application of processes such as optimized culture conditions, and development of the strains with higher yield (e.g. through genetic manipulation), or using low-cost substrates. Objectives In this work, the effects of carbon and nitrogen sources were studied in order to improve the EPS production by the submerged cultivation of Chryseobacterium indologenes MUT.2. Materials and Methods The mesophilic microorganism Chryseobacterium indologenes MUT.2, was grown and maintained in the Luria Bertani agar. The initial basal medium contained: glucose (20 g.L-1), yeast extracts (5 g.L-1), K2HPO4 (6 g.L-1), NaH2PO4 (7 g.L-1), NH4CL (0.7 g.L-1), and MgSO4 (0.5 g.L-1). For evaluating the carbon and nitrogen sources’ effect on the fermentation performance, cultures were prepared in 500 mL flasks filled with 300 mL of the medium. The single-factor experiments based on statistics was employed to evaluate and optimize the carbon and nitrogen sources for EPS production in the liquid culture medium of Chryseobacterium indologenes MUT.2. Results The preferred carbon-sources, sucrose and glucose, commonly gave the highest EPS production of 8.32 and 6.37 g.L-1, respectively, and the maximum EPS production of 8.87 g.L-1 was achieved when glutamic acid (5 g.L-1) was employed as the nitrogen source. Conclusions In this work, the culture medium for production of EPS by Chryseobacterium indologenes MUT.2 was optimized. Compared to the basal culture medium in shake-flasks and stirred tank bioreactor, the use of optimized culture medium has resulted in a 53% and 73% increase in the EPS production, respectively. PMID:28959321

  11. Computational modeling and molecular imprinting for the development of acrylic polymers with high affinity for bile salts.

    PubMed

    Yañez, Fernando; Chianella, Iva; Piletsky, Sergey A; Concheiro, Angel; Alvarez-Lorenzo, Carmen

    2010-02-05

    This work has focused on the rational development of polymers capable of acting as traps of bile salts. Computational modeling was combined with molecular imprinting technology to obtain networks with high affinity for cholate salts in aqueous medium. The screening of a virtual library of 18 monomers, which are commonly used for imprinted networks, identified N-(3-aminopropyl)-methacrylate hydrochloride (APMA.HCl), N,N-diethylamino ethyl methacrylate (DEAEM) and ethyleneglycol methacrylate phosphate (EGMP) as suitable functional monomers with medium-to-high affinity for cholic acid. The polymers were prepared with a fix cholic acid:functional monomer mole ratio of 1:4, but with various cross-linking densities. Compared to polymers prepared without functional monomer, both imprinted and non-imprinted microparticles showed a high capability to remove sodium cholate from aqueous medium. High affinity APMA-based particles even resembled the performance of commercially available cholesterol-lowering granules. The imprinting effect was evident in most of the networks prepared, showing that computational modeling and molecular imprinting can act synergistically to improve the performance of certain polymers. Nevertheless, both the imprinted and non-imprinted networks prepared with the best monomer (APMA.HCl) identified by the modeling demonstrated such high affinity for the template that the imprinting effect was less important. The fitting of adsorption isotherms to the Freundlich model indicated that, in general, imprinting increases the population of high affinity binding sites, except when the affinity of the functional monomer for the target molecule is already very high. The cross-linking density was confirmed as a key parameter that determines the accessibility of the binding points to sodium cholate. Materials prepared with 9% mol APMA and 91% mol cross-linker showed enough affinity to achieve binding levels of up to 0.4 mmol g(-1) (i.e., 170 mg g(-1)) under flow (1 mL min(-1)) of 0.2 mM sodium cholate solution. Copyright 2009 Elsevier B.V. All rights reserved.

  12. 48 CFR 5.501 - Definitions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ..., means any single message prepared for placement in communication media, regardless of the number of... professional journal, or any other printed medium, or (2) the broadcasting of an advertisement over radio or...

  13. Properties of Polyhydroxyalkanoate Granules and Bioemulsifiers from Pseudomonas sp. and Burkholderia sp. Isolates Growing on Glucose.

    PubMed

    Sacco, Laís Postai; Castellane, Tereza Cristina Luque; Lopes, Erica Mendes; de Macedo Lemos, Eliana Gertrudes; Alves, Lúcia Maria Carareto

    2016-03-01

    A Burkholderia and Pseudomonas species designated as AB4 and AS1, respectively, were isolated from soil containing decomposing straw or sugar cane bagasse collected from Brazil. This study sought to evaluate the capacities of culture media, cell-free medium, and crude lysate preparations (containing PHB inclusion bodies) from bacterial cell cultures to stabilize emulsions with several hydrophobic compounds. Four conditions showed good production of bioemulsifiers (E24 ≥ 50 %), headed by substantially cell-free media from bacterial cell cultures in which bacterial isolates from Burkholderia sp. strain AB4 and Pseudomonas sp. strain AS1 were grown. Our results revealed that the both isolates (AB4 and AS1 strains) exhibited high emulsification indices (indicating usefulness in bioremediation) and good stabilities.

  14. Large Area Few Layers Hexagonal Boron Nitride Prepared by Quadrupole Field Aided Exfoliation.

    PubMed

    Hanlun, Lu; Rong, Min Zhi; Zhang, Ming Qiu

    2018-01-16

    A quadrupole electric field mediated exfoliation method is proposed to convert micron sized hexagonal boron nitride (hBN) powders into few layers hexagonal boron nitride nano-sheets (h-BNNS). Under the optimum conditions (400 Hz, 40 V, 32μg/mL, sodium deoxycholate, TAE medium), the hBN powders (thickness > 200 nm, horizontal scale ~ 10 μm) are successfully exfoliated into 0.5-4 nm (1-10 layers) thick h-BNNS with the same horizontal scale. Dynamic laser scattering (DLS) and atomic force microscope (AFM) statistics show that the yield is 47.6 % (for the portion with the thickness of 0.5-6 nm), and all of the vertical sizes are reduced to smaller than 18 nm (45 layers). © 2018 IOP Publishing Ltd.

  15. Dependence of the electronic absorption spectra of aqueous solutions of iodine monochloride on the conditions of dilution and storage time

    NASA Astrophysics Data System (ADS)

    Klyubin, V. V.; Klyubina, K. A.; Makovetskaya, K. N.

    2017-04-01

    The electronic absorption spectra of aqueous solutions of iodine monochloride ICl are studied. The spectra of as-prepared solutions display the absorption band associated with hydrated ICl molecules. An additional band indicating that molecular iodine was formed in the solution emerges in the spectrum as dissolution takes place. Only the band belonging to iodine monochloride remains in the absorption spectra, and no additional bands appear after chloride anions Cl- are added to the solution. The absorption spectrum becomes more complex when ICl is dissolved in an alkaline medium. The band belonging to molecular iodine emerges in the spectra at low alkali concentrations, while being transformed to other shorter-wavelength bands at high alkali concentrations (pH ≥ 12).

  16. A study on the swelling behavior of poly(acrylic acid) hydrogels obtained by electron beam crosslinking

    NASA Astrophysics Data System (ADS)

    Sheikh, N.; Jalili, L.; Anvari, F.

    2010-06-01

    Poly(acrylic acid) (PAA) hydrogels were prepared by using electron beam (EB) crosslinking of PAA homopolymer from its aqueous solutions. The swelling behavior of the hydrogels was studied as a function of the concentration of PAA solution, radiation dose, pH of the swelling medium and swelling time. Also the environmental pH effect on the water diffusion mode into hydrogels was investigated. These hydrogels clearly showed pH-sensitive swelling behavior with Fickian type of diffusion in the stomach-like pH medium (pH 1.3) and non-Fickian type in the intestine-like pH medium (pH 6.8).

  17. Polyphosphoester-Camptothecin Prodrug with Reduction-Response Prepared via Michael Addition Polymerization and Click Reaction.

    PubMed

    Du, Xueqiong; Sun, Yue; Zhang, Mingzu; He, Jinlin; Ni, Peihong

    2017-04-26

    Polyphosphoesters (PPEs), as potential candidates for biocompatible and biodegradable polymers, play an important role in material science. Various synthetic methods have been employed in the preparation of PPEs such as polycondensation, polyaddition, ring-opening polymerization, and olefin metathesis polymerization. In this study, a series of linear PPEs has been prepared via one-step Michael addition polymerization. Subsequently, camptothecin (CPT) derivatives containing disulfide bonds and azido groups were linked onto the side chain of the PPE through Cu(I)-catalyzed azidealkyne cyclo-addition "click" chemistry to yield a reduction-responsive polymeric prodrug P(EAEP-PPA)-g-ss-CPT. The chemical structures were characterized by nuclear magnetic resonance spectroscopy, gel permeation chromatography, Fourier transform infrared, ultraviolet-visible spectrophotometer, and high performance liquid chromatograph analyses, respectively. The amphiphilic prodrug could self-assemble into micelles in aqueous solution. The average particle size and morphology of the prodrug micelles were measured by dynamic light scattering and transmission electron microscopy, respectively. The results of size change under different conditions indicate that the micelles possess a favorable stability in physiological conditions and can be degraded in reductive medium. Moreover, the studies of in vitro drug release behavior confirm the reduction-responsive degradation of the prodrug micelles. A methyl thiazolyl tetrazolium assay verifies the good biocompatibility of P(EAEP-PPA) not only for normal cells, but also for tumor cells. The results of cytotoxicity and the intracellular uptake about prodrug micelles further demonstrate that the prodrug micelles can efficiently release CPT into 4T1 or HepG2 cells to inhibit the cell proliferation. All these results show that the polyphosphoester-based prodrug can be used for triggered drug delivery system in cancer treatment.

  18. Conditioned medium as a strategy for human stem cells chondrogenic differentiation.

    PubMed

    Alves da Silva, M L; Costa-Pinto, A R; Martins, A; Correlo, V M; Sol, P; Bhattacharya, M; Faria, S; Reis, R L; Neves, Nuno M

    2015-06-01

    Paracrine signalling from chondrocytes has been reported to increase the synthesis and expression of cartilage extracellular matrix (ECM) by stem cells. The use of conditioned medium obtained from chondrocytes for stimulating stem cells chondrogenic differentiation may be a very interesting alternative for moving into the clinical application of these cells, as chondrocytes could be partially replaced by stem cells for this type of application. In the present study we aimed to achieve chondrogenic differentiation of two different sources of stem cells using conditioned medium, without adding growth factors. We tested both human bone marrow-derived mesenchymal stem cells (hBSMCs) and human Wharton's jelly-derived stem cells (hWJSCs). Conditioned medium obtained from a culture of human articular chondrocytes was used to feed the cells during the experiment. Cultures were performed in previously produced three-dimensional (3D) scaffolds, composed of a blend of 50:50 chitosan:poly(butylene succinate). Both types of stem cells were able to undergo chondrogenic differentiation without the addition of growth factors. Cultures using hWJSCs showed significantly higher GAGs accumulation and expression of cartilage-related genes (aggrecan, Sox9 and collagen type II) when compared to hBMSCs cultures. Conditioned medium obtained from articular chondrocytes induced the chondrogenic differentiation of MSCs and ECM formation. Obtained results showed that this new strategy is very interesting and should be further explored for clinical applications. Copyright © 2013 John Wiley & Sons, Ltd.

  19. Thermally conductive polymers

    NASA Technical Reports Server (NTRS)

    Byrd, N. R.; Jenkins, R. K.; Lister, J. L. (Inventor)

    1971-01-01

    A thermally conductive polymer is provided having physical and chemical properties suited to use as a medium for potting electrical components. The polymer is prepared from hydroquinone, phenol, and formaldehyde, by conventional procedures employed for the preparation of phenol-formaldehyde resins. While the proportions of the monomers can be varied, a preferred polymer is formed from the monomers in a 1:1:2.4 molar or ratio of hydroquinone:phenol:formaldehyde.

  20. [Occupational and non-occupational factors influencing health state of small and medium business workers].

    PubMed

    Fasikov, R M; Khuzhakhmetova, I B; Stepanov, E G

    2010-01-01

    Complex study of work conditions and health parameters of workers engaged into small and medium business proved that preserved and better health of these workers, prevention of occupational and occupationally mediated diseases necessitate federal and regional complex system of measures including legal basis, database on work conditions and their influence on small and medium business workers' health, occupational medicine training for employers and employees, more active involvement of medical institutioins into screening for occupational diseases.

  1. Exosomes of human placenta-derived mesenchymal stem cells stimulate angiogenesis.

    PubMed

    Komaki, Motohiro; Numata, Yuri; Morioka, Chikako; Honda, Izumi; Tooi, Masayuki; Yokoyama, Naoki; Ayame, Hirohito; Iwasaki, Kengo; Taki, Atsuko; Oshima, Noriko; Morita, Ikuo

    2017-10-03

    The therapeutic potential of mesenchymal stem cells (MSCs) may be attributed partly to humoral factors such as growth factors, cytokines, and chemokines. Human term placental tissue-derived MSCs (PlaMSCs), or conditioned medium left over from cultures of these cells, have been reported to enhance angiogenesis. Recently, the exosome, which can transport a diverse suite of macromolecules, has gained attention as a novel intercellular communication tool. However, the potential role of the exosome in PlaMSC therapeutic action is not well understood. The purpose of this study was to evaluate PlaMSC-derived exosome angiogenesis promotion in vitro and in vivo. MSCs were isolated from human term placental tissue by enzymatic digestion. Conditioned medium was collected after 48-h incubation in serum-free medium (PlaMSC-CM). Angiogenic factors present in PlaMSC-CM were screened by a growth factor array. Exosomes were prepared by ultracentrifugation of PlaMSC-CM, and confirmed by transmission electron microscopy, dynamic light scattering, and western blot analyses. The proangiogenic activity of PlaMSC-derived exosomes (PlaMSC-exo) was assessed using an endothelial tube formation assay, a cell migration assay, and reverse transcription-PCR analysis. The in-vivo angiogenic activity of PlaMSC-exo was evaluated using a murine auricle ischemic injury model. PlaMSC-CM contained both angiogenic and angiostatic factors, which enhanced endothelial tube formation. PlaMSC-exo were incorporated into endothelial cells; these exosomes stimulated both endothelial tube formation and migration, and enhanced angiogenesis-related gene expression. Laser Doppler blood flow analysis showed that PlaMSC-exo infusion also enhanced angiogenesis in an in-vivo murine auricle ischemic injury model. PlaMSC-exo enhanced angiogenesis in vitro and in vivo, suggesting that exosomes play a role in the proangiogenic activity of PlaMSCs. PlaMSC-exo may be a novel therapeutic approach for treating ischemic diseases.

  2. High-oil-load encapsulation of medium-chain triglycerides and D-limonene mixture in modified starch by spray drying.

    PubMed

    Paramita, Vita; Furuta, Takeshi; Yoshii, Hidefumi

    2012-02-01

    Oil mixtures of medium-chain triglycerides (MCT) and D-limonene in mixing ratios from 10 to 100 wt% were encapsulated in modified starch (wall material) by spray drying to produce oil-rich powders. The oil load (mass ratio of oil mixture to wall material) of the infeed emulsion markedly influenced the properties of the infeed liquid and the characteristics of the resulting powder. The viscosity of the infeed liquid and the particle size of the powder exponentially decreased with increasing oil load, while the emulsion droplet size in the infeed liquid increased. In addition, retention of D-limonene during spray drying also decreased markedly with increasing oil load. Irrespective of the different oil loads and concentrations of the wall material, D-limonene retention was well correlated with the emulsion droplet diameter of the infeed liquid. The encapsulation efficiency of the oil mixture exhibited a maximum value (almost 100%) at an oil load between 0.5 and 1.0, before decreasing at higher oil loads. At an oil load of 2.0, the encapsulation efficiency of D-limonene was reduced to almost zero, while around 40% of the initial MCT was encapsulated in the powder. The increase in oil load also led to increased amounts of surface oil of MCT and D-limonene in the resulting powder due to the increasing emulsion droplet diameter of the infeed liquids. This study proposes the microencapsulation of medium-chain triglycerides under high-oil-load conditions by spray drying. The powders prepared by this process provide significant benefits in terms of rapid energy conversion after consumption without accumulation in the body. Important quality factors of the powder products such as the encapsulation efficiency and the amount of surface oil were examined to understand the optimum process conditions for spray drying. © 2012 Institute of Food Technologists®

  3. 40 CFR Table 1 to Subpart Hhh of... - Emission Limits for Small Rural, Small, Medium, and Large HMIWI

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ..., Medium, and Large HMIWI 1 Table 1 to Subpart HHH of Part 62 Protection of Environment ENVIRONMENTAL... 62—Emission Limits for Small Rural, Small, Medium, and Large HMIWI Pollutant Units (7 percent oxygen, dry basis at standard conditions) Emission limits HMIWI size Small rural Small Medium Large...

  4. 40 CFR Table 1 to Subpart Hhh of... - Emission Limits for Small Rural, Small, Medium, and Large HMIWI

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., Medium, and Large HMIWI 1 Table 1 to Subpart HHH of Part 62 Protection of Environment ENVIRONMENTAL... 62—Emission Limits for Small Rural, Small, Medium, and Large HMIWI Pollutant Units (7 percent oxygen, dry basis at standard conditions) Emission limits HMIWI size Small rural Small Medium Large...

  5. 40 CFR Table 1 to Subpart Hhh of... - Emission Limits for Small Rural, Small, Medium, and Large HMIWI

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., Medium, and Large HMIWI 1 Table 1 to Subpart HHH of Part 62 Protection of Environment ENVIRONMENTAL... 62—Emission Limits for Small Rural, Small, Medium, and Large HMIWI Pollutant Units (7 percent oxygen, dry basis at standard conditions) Emission limits HMIWI size Small rural Small Medium Large...

  6. Callus induction and flavonoid production on the immature seed of Stelechocarpus burahol

    NASA Astrophysics Data System (ADS)

    Habibah, N. A.; Moeljopawiro, S.; Dewi, K.; Indrianto, A.

    2018-03-01

    Stelechocarpus burahol [(Bl.) Hook. f. & Th.] is one of the medicinal plants. In vitro callus induction studies on S. burahol were carried out to determine phytohormone requirement for optimum callus induction. Immature seed explants were cultured on MS medium by adding different kinds and different concentrations of plant growth regulators (picloram and 2,4-D) under light and dark conditions. The results showed that callus formation was initiated on the 18,50th to the 55th days. The best condition for optimum callus induction was found on MS medium, which was supplemented with 7.5 mg/L picloram and was maintained in the dark condition. The callus induction varied from 60% to 100%. The callus that produced the highest flavonoid was grown on the medium with the addition of 10 mg/L of 2,4-D. In conclusion, the results represented a suitable medium for S.burahol callus induction.

  7. 15 CFR 1180.4 - Preparing a product for transfer.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... SCIENTIFIC, TECHNICAL AND ENGINEERING INFORMATION TO THE NATIONAL TECHNICAL INFORMATION SERVICE § 1180.4... medium; (3) In the case of software, be accompanied by relevant documentation, such as operating manuals...

  8. Electrophoretic analysis of the major polypeptides of human erythrocyte membranes prepared by low and high osmolarity haemolysis.

    PubMed

    Zail, S S; Hoek, V D

    1975-04-16

    Human erythrocyte membranes were prepared in three ways: washing in hypotonic Tris buffer, pH 7.6, by lysis in isotonic Tris buffer pH 7.6 after incubation at 37 degrees C for 2 hours and by ultrasonication in an isotonic medium, pH 7.6. Analysis of the major polypeptides of the erythrocyte membranes by sodium dodecylsulphate polyacrylamide gel electrophoresis revealed a selective depletion of a major polypeptide representing glyceraldehyde-3-phosphate dehydrogenase in the membranes prepared by high osmolarity lysis. The pattern of seperation of the remaining polypeptides was identical in the 3 different membrane preparations.

  9. Preparation of 14C-Labeled Sterigmatocystin in Liquid Media

    PubMed Central

    Hsieh, Dennis P. H.; Yang, Susie L.

    1975-01-01

    14C-labeled sterigmatocystin was prepared from surface cultures of Aspergillus versicolor A-18074 maintained in liquid media by multiple additions of [1-14C]acetate to the cultures. The highest yield of 7.75 mg/10 ml was found with a sucrose-asparagine-ammonium medium in which more than 3% of the radioactivity of the added [1-14C]acetate was recovered in the purified [ring-14C] sterigmatocystin. The method offers an easy way to prepare 14C-labeled sterigmatocystin for studies of this mycotoxin. PMID:1110489

  10. European Seminar on the Development of the Competencies of Trainers and Organizers of Training Programmes for SMES (Berlin, West Germany, June 7-8, 1989). CEDEFOP Flash 7/89.

    ERIC Educational Resources Information Center

    Melis, A.

    This document summarizes the proceedings of a 2-day conference on training in small and medium businesses in the European Economic Community. Some of the topics discussed by the speakers include distance learning and the importance of training for owners and managers of small and medium enterprises. A series of guides that have been prepared to…

  11. [Alterations in the metabolism of cornmeal epithelium during medium-term storage (author's transl)].

    PubMed

    Schmidt-Martens, F W; Hennighausen, U; Wirz, K; Teping, C

    1977-08-08

    Freshly prepared bovine corneas were stored in medium TC 199 with penicillin and fetal calf serum at +4 degrees C over a storage period of 168h. Every 24h, the levels of glucose, lactate, and pyruvate in the corneal epithelium were estimated. Also the glucose levels in the corneal epithelium and stroma were compared at the same time intervals. Furthermore, alterations in the enzyme pattern of the epithelial cells during storage were observed.

  12. Determination of physicochemical properties and degradation kinetics of triamcinolone acetonide palmitate in vitro.

    PubMed

    Peng, Cuilian; Liu, Cong; Tang, Xing

    2010-12-01

    Triamcinolone acetonide palmitate (TAP) is a lipophilic prodrug of triamcinolone acetonide (TAA) to improve the insoluble TAA physicochemical properties for the preparation of emulsions. This investigation has focused on the preformulation study of TAP, including its physicochemical properties and hydrolysis kinetics in vitro. The solubility of TAP in medium-chain triglyceride is about twice greater than that in soybean oil (long-chain triglyceride) (19.17 versus 9.55 mg/g) at 25°C, and in all investigated cases, lecithin (80, 160, and 240 mg/g) as solubilizer provided increased solubility of drugs in medium-chain triglyceride and long-chain triglyceride, whereas the maximum water solubility of TAP was 0.10 μg/mL. The partition coefficient (log P) of TAP was 5.79 irrespective of the pH conditions. The hydrolysis of TAP followed pseudo-first-order kinetics in aqueous solutions, and the stable pH range was from pH 5.0 to 9.0. The in vitro enzymolysis kinetics of TAP in rat plasma and liver homogenate was evaluated by measuring the decrease of TAP as well as the increase of TAA at 37°C for 96 hours. The results demonstrated that the TAP may be hydrolyzed mainly by rat plasma esterase and, to a minor extent, by liver esterase, and the hydrolysis half-life of TAP in 100% rat plasma was 17.53 ± 6.85 hours at pH 7.4. All these results indicated that TAP had successfully obtained higher lipid-soluble property for the preparation of intravenous emulsion and may be an effective prodrug for sustained release of TAA in vivo.

  13. Matrix-assisted laser desorption/ionization mass spectrometric analysis of poly(3,4-ethylenedioxythiophene) in solid-state dye-sensitized solar cells: comparison of in situ photoelectrochemical polymerization in aqueous micellar and organic media.

    PubMed

    Zhang, Jinbao; Ellis, Hanna; Yang, Lei; Johansson, Erik M J; Boschloo, Gerrit; Vlachopoulos, Nick; Hagfeldt, Anders; Bergquist, Jonas; Shevchenko, Denys

    2015-04-07

    Solid-state dye-sensitized solar cells (sDSCs) are devoid of such issues as electrolyte evaporation or leakage and electrode corrosion, which are typical for traditional liquid electrolyte-based DSCs. Poly(3,4-ethylenedioxythiophene) (PEDOT) is one of the most popular and efficient p-type conducting polymers that are used in sDSCs as a solid-state hole-transporting material. The most convenient way to deposit this insoluble polymer into the dye-sensitized mesoporous working electrode is in situ photoelectrochemical polymerization. Apparently, the structure and the physicochemical properties of the generated conducting polymer, which determine the photovoltaic performance of the corresponding solar cell, can be significantly affected by the preparation conditions. Therefore, a simple and fast analytical method that can reveal information on polymer chain length, possible chemical modifications, and impurities is strongly required for the rapid development of efficient solar energy-converting devices. In this contribution, we applied matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) for the analysis of PEDOT directly on sDSCs. It was found that the PEDOT generated in aqueous micellar medium possesses relatively shorter polymeric chains than the PEDOT deposited from an organic medium. Furthermore, the micellar electrolyte promotes a transformation of one of the thiophene terminal units to thiophenone. The introduction of a carbonyl group into the PEDOT molecule impedes the growth of the polymer chain and reduces the conductivity of the final polymer film. Both the simplicity of sample preparation (only application of the organic matrix onto the solar cell is needed) and the rapidity of analysis hold the promise of making MALDI MS an essential tool for the physicochemical characterization of conducting polymer-based sDSCs.

  14. Exploring the sensitivity of the zone of inhibition test for leachable biocides from paper and board food contact materials, and improvements thereof.

    PubMed

    Castle, L; Kelly, J; Jickells, S M; Johns, S M; Mountfort, K A

    2012-01-01

    The zone of inhibition method to test the release of biocides from paper and board food contact materials was evaluated. The method tests the paper by placing a small specimen directly onto culture plates of Bacillus subtilis and Aspergillus niger. The principle is that any extractable biocide will diffuse from the paper into the surrounding nutrient medium and so inhibit growth of the microorganism in the vicinity. The test was found to have insufficient sensitivity for assuring food safety, where detection limits for migration at or below the mg l(-1) (parts per million) level are needed. Also, the test does not mimic the actual or foreseeable conditions of use since most paper/board materials are not intended for direct contact with an aqueous medium for up to 3 days at 30°C (B. subtilis) or 25°C (A. niger), which are the incubation conditions used. The sensitivity of the test was increased approximately 100-fold by preparing a concentrated extract of the paper to be tested and applying this extract to the assay via a blank paper carrier. This was done using methanol as a good solvent for most biocides, as a proof of principle. Other solvents or food simulants could be used to mimic the conditions of use intended for the particular paper/board samples under examination, e.g. contact with dry, fatty, aqueous or acidic foods, hot or cold. Twenty-four plain (unconverted) paper and board samples and 100 food packaging samples were evaluated using the modified procedure. The results revealed that the method has been developed to the stage where background cytotoxic action of normal paper constituents gives a weak response. Unlike the original method, therefore, the modified method with its improved sensitivity and the facility to link with the intended food contact conditions may be considered a suitable bioassay screening test to complement chemical analysis of paper/board for composition and migration.

  15. An Investigation into the Suitability of Sulfate-Reducing Bacteria as Models for Martian Forward Contamination

    NASA Astrophysics Data System (ADS)

    Silver, Maxwell M. W.

    The NASA Planetary Protection policy requires interplanetary space missions do not compromise the target body for a current or future scientific investigation and do not pose an unacceptable risk to Earth, including biologic materials. Robotic missions to Mars pose a risk to planetary protection in the forms of forward and reverse contamination. To reduce these risks, a firm understanding of microbial response to Mars conditions is required. Sulfate-reducing bacteria are prime candidates for potential forward contamination on Mars. Understanding the potential for forward-contamination of sulfate-reducers on Mars calls for the characterization of sulfate-reducers under Mars atmosphere, temperature, and sulfate-brines. This study investigated the response of several sulfate-reducing bacteria, including spore formers and psychrophiles. The psychrophile Desulfotalea psychrophila was found to inconsistently survive positive control lab conditions, attributed to an issue shipping pure cultures. Desulfotomaculum arcticum, a spore-forming mesophilic sulfate-reducer, and Desulfuromusa ferrireducens, an iron and sulfate-reducer, were metabolically active under positive control lab conditions with complex and minimal growth medium. A wastewater treatment sulfate-reducing bacteria (SRB) isolate was subjected to sulfate + growth-medium solutions of varied concentrations (0.44 & 0.55% wt. SO42-). The wastewater SRB displayed higher cellular light-absorbance levels at delayed rates in 0.55% sulfate solutions, suggesting a greater total culture reproduction, but with increased lag time. Additional SRB were isolated from marine sediments, subjected to a shock pressure of 8.73 GPa, and returned to ideal conditions. The sulfate-concentration patterns in the impacted SRB culture suggests a destruction of culture occurred somewhere during the preparation process. The response of SRB in this investigation to Ca and Na sulfate-brines suggests that Martian sulfate deposits offer a viable energy sink to terrestrial microorganisms, and the studied SRB are capable of replication at reduced water-activity. Further investigation (i.e. sulfate cations and concentrations, temperature, pressure, etc.) may identify Martian locations at risk to forward contamination.

  16. Mechanisms for the target patterns formation in a stochastic bistable excitable medium

    NASA Astrophysics Data System (ADS)

    Verisokin, Andrey Yu.; Verveyko, Darya V.; Postnov, Dmitry E.

    2018-04-01

    We study the features of formation and evolution of spatiotemporal chaotic regime generated by autonomous pacemakers in excitable deterministic and stochastic bistable active media using the example of the FitzHugh - Nagumo biological neuron model under discrete medium conditions. The following possible mechanisms for the formation of autonomous pacemakers have been studied: 1) a temporal external force applied to a small region of the medium, 2) geometry of the solution region (the medium contains regions with Dirichlet or Neumann boundaries). In our work we explore the conditions for the emergence of pacemakers inducing target patterns in a stochastic bistable excitable system and propose the algorithm for their analysis.

  17. FIBER AND INTEGRATED OPTICS. OTHER TOPICS IN QUANTUM ELECTRONICS: Determination of the saturation parameter in a fast-flow CO2 active medium

    NASA Astrophysics Data System (ADS)

    Lebedev, V. F.; Lysikov, A. Yu; Ryazanova, A. V.; Shalygin, S. V.

    1990-05-01

    A diagnostic method was developed for determination of such an important characteristic of a CO2 active medium as the energy stored in the vibrational degrees of freedom of molecules, which can be determined directly from the experimentally measured powers of probe beams without the need for any additional information on the medium and without any numerical calculations. Measurements were carried out using a real fast-flow system, which satisfied the conditions of validity of the proposed diagnostic method, and the correctness of the approach was confirmed. The results obtained demonstrated the efficiency of the excitation of the medium when the pump conditions were varied.

  18. [Method of culturing microorganisms at constant concentrations of the nutrient components].

    PubMed

    Markvichev, N S; Manakov, M N

    1985-01-01

    A method for batch cultivation of microorganisms in a flow medium is described, characterized by slight changes in concentrations of medium components in time and by the absence of products of vital activity of microorganisms in the fermentation medium. The conditions are achieved due to application of a fermentation installation with a microfiltrative membrane that separates the cells of cultivated microorganisms from the culture fluid and due to increasing the flow rate to a value at which the inlet and outlet concentrations of the medium components are almost equal. The cells of cultivated microorganisms under such conditions remain in the fermentation medium volume. The system was called "Ekostat". If the process is performed in "Ekostat" system, a positive deviation from the logarithmic law is observed for the growth rate of the yeast Candida utilis VSB-651 on ethanol cultivation.

  19. Corrosive and cytotoxic properties of compact specimens and microparticles of Ni-Cr dental alloy.

    PubMed

    Ristic, Ljubisa; Vucevic, Dragana; Radovic, Ljubica; Djordjevic, Snezana; Nikacevic, Milutin; Colic, Miodrag

    2014-04-01

    Nickel-chromium (Ni-Cr) dental alloys have been widely used in prosthodontic practice, but there is a permanent concern about their biocompatibility due to the release of metal ions. This is especially important when Ni-Cr metal microparticles are incorporated into gingival tissue during prosthodontic procedures. Therefore, the aim of this study was to examine and compare the corrosion and cytotoxic properties of compact specimens and microparticles of Ni-Cr dental alloy. Ni-Cr alloy, Remanium CSe bars (4 mm diameter), were made by the standard casting method and then cut into 0.5-mm-thick disks. Metal particles were obtained by scraping the bars using a diamond instrument for crown preparation. The microstructure was observed by an optical microscope. Quantitative determination and morphological and dimensional characterization of metal particles were carried out by a scanning electron microscope and Leica Application Suite software for image analysis. Corrosion was studied by conditioning the alloy specimens in the RPMI 1640 medium, containing 10% fetal calf serum in an incubator with 5% CO2 for 72 hours at 37°C. Inductively coupled plasma-optical emission spectrometry was used to assess metal ion release. The cytotoxity of conditioning medium (CM) was investigated on L929 cells using an MTT test. One-way ANOVA was used for statistical analysis. After casting, the microstructure of the Remanium CSe compact specimen composed of Ni, Cr, Mo, Si, Fe, Al, and Co had a typical dendritic structure. Alloy microparticles had an irregular shape with a wide size range: from less than 1 μm to more than 100 μm. The release of metal ions, especially Ni and Mo from microparticles, was significantly higher, compared to the compact alloy specimen. The CM prepared from compact alloy was not cytotoxic at any tested dilutions, whereas CM from alloy microparticles showed dose-dependent cytotoxicity (90% CM and 45% CM versus control; p < 0.005). Ni-Cr microparticles showed less corrosion resistance and lower biocompatibility than compact alloy. This could affect health on long-term exposure, especially in sensitized individuals. © 2013 by the American College of Prosthodontists.

  20. Platelet concentrates for transfusion-metabolic and storage aspects.

    PubMed

    Farrugia, A

    1994-01-01

    Transfusion of platelets concentrated from donated blood is an established therapeutic modality in clinical medicine. Over the past 25 years much effort has gone into optimising the conditions for the collection, preparation and storage of platelets for transfusion. Despite significant advances, platelet production is still a costly process requiring a dedicated environment and the use of specially formulated plastic storage containers. A progressive lesion over storage limits the shelf life and the availability of donated platelets, while the need to store platelets in the donor's autologous plasma also results in a loss of valuable fresh plasma for fractionation. Recent studies have addressed the issues of platelet quality and plasma economy by examining the possibility of storing platelets in a synthetic medium. Platelets stored in a variety of crystalloid solutions have been shown to retain in vitro and in vivo properties equivalent or superior to platelets stored in autologous donor plasma. Some additional insight has been gained on the metabolic patterns of stored platelets. In particular, studies have shown that, under these conditions, platelets are unable to oxidise dextrose to any significant extent, and that dextrose is invariably broken down to lactate, irrespective of the oxygen tensions in the platelet's environment. This in turn leads to the metabolic lesion of platelet storage, whereby low pH results in loss of platelet viability. Platelets stored in synthetic dextrose-free media are capable of maintaining aerobic ATP generation, and acetate-a component of many media studied-has been shown to be metabolised by platelets. Similarly, platelets prepared from blood collected into a dextrose-free anticoagulant have satisfactory properties both when suspended in autologous plasma or in a dextrose-free synthetic medium. The requirements for storage in special, high gas-permeable, containers, and for constant agitation during storage, were both found to be unnecessary when dextrose was excluded from the platelet's environment. These developments suggest that manipulation of the platelet's metabolic pattern during blood bank storage may allow significant benefits in plasma economy as well as in decreasing the cost of platelet delivery to patients.

  1. CD109 is a component of exosome secreted from cultured cells

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

    Sakakura, Hiroki; Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, Nagoya; Mii, Shinji

    Exosomes are 50–100-nm-diameter membrane vesicles released from various types of cells. Exosomes retain proteins, mRNAs and miRNAs, which can be transported to surrounding cells. CD109 is a glycosylphosphatidylinositol-anchored glycoprotein, and is released from the cell surface to the culture medium in vitro. Recently, it was reported that secreted CD109 from the cell surface downregulates transforming growth factor-β signaling in human keratinocytes. In this study, we revealed that CD109 is a component of the exosome in conditioned medium. FLAG-tagged human CD109 (FLAG-CD109) in conditioned medium secreted from HEK293 cells expressing FLAG-CD109 (293/FLAG-CD109) was immunoprecipitated with anti-FLAG affinity gel, and the co-precipitated proteins weremore » analyzed by mass spectrometry and western blotting. Exosomal proteins were associated with CD109. We revealed the presence of CD109 in exosome fractions from conditioned medium of 293/FLAG-CD109. Moreover, the localization of CD109 in the exosome was demonstrated using immuno-electron microscopy. When we used HEK293 cells expressing FLAG-tagged truncated CD109, which does not contain the C-terminal region, the association of truncated CD109 with exosomes was not detected in conditioned medium. These findings indicate that CD109 is an exosomal protein and that the C-terminal region of CD109 is required for its presence in the exosome. - Highlights: • CD109 is an exosomal protein. • The C-terminal region of CD109 is required for its presence in the exosome. • Part of the secreted CD109 is present in the exosome-free fraction in the conditioned medium.« less

  2. Epidermal regulation of dermal fibroblast activity.

    PubMed

    Garner, W L

    1998-07-01

    Although the association between delayed burn wound healing and subsequent hypertrophic scar formation is well-established, the mechanism for this relationship is unknown. Unhealed burn wounds lack an epidermis, suggesting a possible regulatory role for the epidermis in controlling dermal fibroblast matrix synthesis. Therefore, we examined the effect of epidermal cells and media conditioned by epidermal cells on fibroblast collagen synthesis and replication. Purified fibroblast and keratinocyte cell strains were developed from discarded normal adult human skin. Conditioned media were created by incubation of cytokine-free and serum-free medium with either confluent fibroblast or keratinocyte cultures for 18 hours (n = 3). Nearly confluent fibroblast cultures were exposed for 48 hours to graded concentrations of either unconditioned medium (control), conditioned medium, or varying numbers of keratinocytes. Replication was quantified by the incorporation of 3H-thymidine. Collagen synthesis was measured by the incorporation of 3H-proline into collagenase-sensitive protein. Data were compared using analysis of variance (ANOVA) and linear regression. Keratinocyte conditioned medium induced a significant increase in replication (n = 3) (p = 0.004) and a decrease in collagen synthesis (n = 6) (p < 0.001). In contrast, neither fibroblast conditioned medium nor control medium had an effect on fibroblast replication or collagen synthesis. Co-culture of fibroblast with a graded number of keratinocytes similarly decreased collagen synthesis (n = 6) (p < 0.001). Dermal fibroblast collagen synthesis appears to be regulated by a soluble keratinocyte product. This result suggests a mechanism for the clinical observation that unhealed burn wounds, which lack the epidermis, demonstrate excess collagen production and scar. Clinical strategies to decrease hypertrophic scar should include an attempt at early wound closure with skin grafting or the application of cultured epithelial autografts.

  3. Lung cells support osteosarcoma cell migration and survival.

    PubMed

    Yu, Shibing; Fourman, Mitchell Stephen; Mahjoub, Adel; Mandell, Jonathan Brendan; Crasto, Jared Anthony; Greco, Nicholas Giuseppe; Weiss, Kurt Richard

    2017-01-25

    Osteosarcoma (OS) is the most common primary bone tumor, with a propensity to metastasize to the lungs. Five-year survival for metastatic OS is below 30%, and has not improved for several decades despite the introduction of multi-agent chemotherapy. Understanding OS cell migration to the lungs requires an evaluation of the lung microenvironment. Here we utilized an in vitro lung cell and OS cell co-culture model to explore the interactions between OS and lung cells, hypothesizing that lung cells would promote OS cell migration and survival. The impact of a novel anti-OS chemotherapy on OS migration and survival in the lung microenvironment was also examined. Three human OS cell lines (SJSA-1, Saos-2, U-2) and two human lung cell lines (HULEC-5a, MRC-5) were cultured according to American Type Culture Collection recommendations. Human lung cell lines were cultured in growth medium for 72 h to create conditioned media. OS proliferation was evaluated in lung co-culture and conditioned media microenvironment, with a murine fibroblast cell line (NIH-3 T3) in fresh growth medium as controls. Migration and invasion were measured using a real-time cell analysis system. Real-time PCR was utilized to probe for Aldehyde Dehydrogenase (ALDH1) expression. Osteosarcoma cells were also transduced with a lentivirus encoding for GFP to permit morphologic analysis with fluorescence microscopy. The anti-OS efficacy of Disulfiram, an ALDH-inhibitor previously shown to inhibit OS cell proliferation and metastasis in vitro, was evaluated in each microenvironment. Lung-cell conditioned medium promoted osteosarcoma cell migration, with a significantly higher attractive effect on all three osteosarcoma cell lines compared to basic growth medium, 10% serum containing medium, and NIH-3 T3 conditioned medium (p <0.05). Lung cell conditioned medium induced cell morphologic changes, as demonstrated with GFP-labeled cells. OS cells cultured in lung cell conditioned medium had increased alkaline phosphatase staining. Lung endothelial HULEC-5a cells are attractants for OS cell migration, proliferation, and survival. The SJSA-1 osteosarcoma cell line demonstrated greater metastatic potential than Saos-2 and U-2 cells. ALDH appears to be involved in the interaction between lung and OS cells, and ALP may be a valuable biomarker for monitoring functional OS changes during metastasis.

  4. Accumulation and chemical states of radiocesium by fungus Saccharomyces cerevisiae

    NASA Astrophysics Data System (ADS)

    Ohnuki, Toshihiko; Sakamoto, Fuminori; Kozai, Naofumi; Yamasaki, Shinya; Yu, Qianqian

    2014-05-01

    After accident of Fukushima Daiichi Nuclear Power Plant, the fall-out radiocesium was deposited on the ground. Filamentous fungus is known to accumulate radiocesium in environment, even though many minerals are involved in soil. These facts suggest that fungus affect the migration behavior of radiocesium in the environment. However, accumulation mechanism of radiocesium by fungus is not understood. In the present study, accumulation and chemical states change of Cs by unicellular fungus of Saccharomyces cerevisiae have been studied to elucidate the role of microorganisms in the migration of radiocesium in the environment. Two different experimental conditions were employed; one is the accumulation experiments of radiocesium by S. cerevisiae from the agar medium containing 137Cs and a mineral of zeolite, vermiculite, smectite, mica, or illite. The other is the experiments using stable cesium to examine the chemical states change of Cs. In the former experiment, the cells were grown on membrane filter of 0.45 μm installed on the agar medium. After the grown cells were weighed, radioactivity in the cells was measured by an autoradiography technique. The mineral weight contents were changed from 0.1% to 1% of the medium. In the latter experiment, the cells were grown in the medium containing stable Cs between 1 mM and 10mM. The Cs accumulated cells were analyzed by SEM-EDS and EXAFS. The adsorption experiments of cesium by the cells under resting condition were also conducted to test the effect of cells metabolic activity. Without mineral in the medium, cells of S. cerevisiae accumulated 1.5x103 Bq/g from the medium containing 137Cs of 2.6x102 Bq/g. When mineral was added in the medium, concentration of 137Cs in the cells decreased. The concentration of 137Cs in the cells from the medium containing different minerals were in the following order; smectite, illite, mica > vermiculite > zeolite. This order was nearly the same as the inverse of distribution coefficient of mineral for 137Cs in the medium solution. The concentration of 137Cs in the cells lowered in the medium containing higher mineral content. These results indicate that radiocesium was competively accumulated in the cells with minerals in the soil. Higher concentration of stable Cs was accumulated in the cells in the metabolically active condition than in the resting cells condition. XAFS analyses showed that the k3-weighted extended-XAFS functions and the radial structural function of Cs accumulated by the cells in the metabolically active condition were similar to those in the resting condition, indicating that chemical states of the accumulated Cs were nearly the same between both conditions. These results indicate that the fungus accumulates radiocesium by competitively with minerals in the soils, and performs higher retardation of the migration of Cs in the metabolically active condition than the resting one. A part of this study is the results of "Multidisciplinary investigation on radiocesium fate and transport for safety assessment for interim storage and disposal of heterogeneous waste" carried out under the Initiatives for Atomic Energy Basic and Generic Strategic Research by the Ministry of Education, Culture, Sports, Science and Technology of Japan.

  5. Astrocytes produce an insulin-like neurotrophic factor

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

    Kadle, R.; Suksang, C.; Fellows, R.E.

    1986-05-01

    They have previously reported that survival of dissociated neurons from fetal rat telencephalon plated at low density in serum-free, hormone-free defined medium is enhanced in the presence of insulin. In the absence of insulin a similar effect on neuronal survival is observed if cells are grown in medium conditioned by glial cells. The present study was carried out to characterize the insulin-like neurotrophic activity present in the glial conditioned medium (GLCM). Conditioned medium from confluent cultures of astrogial cells maintained in a serum free defined medium without insulin was collected every two or three days. A 5 to 30kDa fractionmore » of this medium was obtained by filtering it sequentially through YM30 and YM5 membrane filters. Binding of /sup 125/I-insulin to high density neuronal cultures was inhibited 43% by this fraction. Radioimmunoassay for insulin indicated that 1-2 ng of immuno-reactive insulin were present per ml of GLCM. Immunosequestration of the factor by insulin antibodies bound to protein A agarose gel resulted in loss of neurotrophic activity of the 5 to 30 kDa fraction. These results indicate that cultured astrocytes produce a factor immunologically and biochemically similar to insulin. This factor enhances the survival of neurons in culture and may be important for their normal development and differentiation.« less

  6. Adult hippocampus derived soluble factors induce a neuronal-like phenotype in mesenchymal stem cells.

    PubMed

    Rivera, Francisco J; Sierralta, Walter D; Minguell, Jose J; Aigner, Ludwig

    2006-10-02

    Bone marrow-derived mesenchymal stem cells (MSCs) are not restricted in their differentiation fate to cells of the mesenchymal lineage. They acquire a neural phenotype in vitro and in vivo after transplantation in the central nervous system. Here we investigated whether soluble factors derived from different brain regions are sufficient to induce a neuronal phenotype in MSCs. We incubated bone marrow-derived MSCs in conditioned medium (CM) derived from adult hippocampus (HCM), cortex (CoCM) or cerebellum (CeCM) and analyzed the cellular morphology and the expression of neuronal and glial markers. In contrast to muscle derived conditioned medium, which served as control, conditioned medium derived from the different brain regions induced a neuronal morphology and the expression of the neuronal markers GAP-43 and neurofilaments in MSCs. Hippocampus derived conditioned medium had the strongest activity. It was independent of NGF or BDNF; and it was restricted to the neuronal differentiation fate, since no induction of the astroglial marker GFAP was observed. The work indicates that soluble factors present in the brain are sufficient to induce a neuronal phenotype in MSCs.

  7. Aqueous extracts of a Mars analogue regolith that mimics the Phoenix landing site do not inhibit spore germination or growth of model spacecraft contaminants Bacillus subtilis 168 and Bacillus pumilus SAFR-032

    NASA Astrophysics Data System (ADS)

    Nicholson, Wayne L.; McCoy, Lashelle E.; Kerney, Krystal R.; Ming, Douglas W.; Golden, D. C.; Schuerger, Andrew C.

    2012-08-01

    Because Mars is a primary target for life detection and habitability assessment missions, its exploration is also by necessity a Planetary Protection issue. The recent finding of significant levels of perchlorate (ClO4-) in regolith sampled from the Phoenix landing site raises the question of its potential biotoxicity to putative indigenous martian life, microbial forward contaminants from Earth, or future human visitors. To address this issue, an analogue regolith was constructed based on regolith chemistry data from the Phoenix landing site. A Mars Aqueous Regolith Extract (MARE) was prepared from the Phoenix analogue regolith and analyzed by ion chromatography. The MARE contained (mg/L) the cations Na+ (1411 ± 181), Mg2+ (1051 ± 160), Ca2+ (832 ± 125), and K+ (261 ± 29), and the anions SO42-(5911±993), ClO4-(5316±1767), Cl(171±25) and F- (2.0 ± 0.4). Nitrogen-containing species NO3-(773±113) and NO2-(6.9±2.3) were also present as a result of regolith preparation procedures, but their relevance to Mars is at present unknown. The MARE was tested for potential toxic effects on two model spacecraft contaminants, the spore-forming bacteria Bacillus subtilis strain 168 and Bacillus pumilus strain SAFR-032. In B. subtilis, spore germination and initial vegetative growth (up to ˜5 h) was not inhibited in a rich complex medium prepared with the MARE, but growth after 5 h was significantly suppressed in medium prepared using the MARE. Both B. subtilis and B. pumilus exhibited significantly higher rates of spore germination and growth in the MARE vs. DW with no additions (likely due to endogenous spore nutrients), but germination and growth was further stimulated by addition of glucose and a combination of buffered inorganic salts (K2HPO4, KH2PO4, (NH4)2SO4, and MgSO4). The data indicate that the aqueous environment in the regolith from the Phoenix landing site containing high levels of perchlorate does not pose a significant barrier to growth of putative forward contaminants such as B. subtilis and B. pumilus under Earth laboratory conditions.

  8. Changes of the elemental distributions in marine diatoms as a reporter of sample preparation artefacts. A nuclear microscopy application

    NASA Astrophysics Data System (ADS)

    Godinho, R. M.; Cabrita, M. T.; Alves, L. C.; Pinheiro, T.

    2015-04-01

    Studies of the elemental composition of whole marine diatoms cells have high interest as they constitute a direct measurement of environmental changes, and allow anticipating consequences of anthropogenic alterations to organisms, ecosystems and global marine geochemical cycles. Nuclear microscopy is a powerful tool allowing direct measurement of whole cells giving qualitative imaging of distribution, and quantitative determination of intracellular concentration. Major obstacles to the analysis of marine microalgae are high medium salinity and the recurrent presence of extracellular exudates produced by algae to maintain colonies in natural media and in vitro. The objective of this paper was to optimize the methodology of sample preparation of marine unicellular algae for elemental analysis with nuclear microscopy, allowing further studies on cellular response to metals. Primary cultures of Coscinodiscus wailesii maintained in vitro were used to optimize protocols for elemental analysis with nuclear microscopy techniques. Adequate cell preparation procedures to isolate the cells from media components and exudates were established. The use of chemical agents proved to be inappropriate for elemental determination and for intracellular morphological analysis. The assessment of morphology and elemental partitioning in cell compartments obtained with nuclear microscopy techniques enabled to infer their function in natural environment and imbalances in exposure condition. Exposure to metal affected C. wailesii morphology and internal elemental distribution.

  9. Human platelet lysate: Replacing fetal bovine serum as a gold standard for human cell propagation?

    PubMed

    Burnouf, Thierry; Strunk, Dirk; Koh, Mickey B C; Schallmoser, Katharina

    2016-01-01

    The essential physiological role of platelets in wound healing and tissue repair builds the rationale for the use of human platelet derivatives in regenerative medicine. Abundant growth factors and cytokines stored in platelet granules can be naturally released by thrombin activation and clotting or artificially by freeze/thaw-mediated platelet lysis, sonication or chemical treatment. Human platelet lysate prepared by the various release strategies has been established as a suitable alternative to fetal bovine serum as culture medium supplement, enabling efficient propagation of human cells under animal serum-free conditions for a multiplicity of applications in advanced somatic cell therapy and tissue engineering. The rapidly increasing number of studies using platelet derived products for inducing human cell proliferation and differentiation has also uncovered a considerable variability of human platelet lysate preparations which limits comparability of results. The main variations discussed herein encompass aspects of donor selection, preparation of the starting material, the possibility for pooling in plasma or additive solution, the implementation of pathogen inactivation and consideration of ABO blood groups, all of which can influence applicability. This review outlines the current knowledge about human platelet lysate as a powerful additive for human cell propagation and highlights its role as a prevailing supplement for human cell culture capable to replace animal serum in a growing spectrum of applications. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. The components of an α-glycerophosphate cycle and their relation to oxidative metabolism in the lens

    PubMed Central

    Griffiths, M. H.

    1966-01-01

    1. The concentration of ATP in a lens brei is maintained when the brei is incubated in oxygen with α-glycerophosphate. Lack of α-glycerophosphate or incubation in nitrogen causes the concentration to decrease. α-Glycerophosphate has some effect under anaerobic conditions but this is not sufficient to account for the maintenance in oxygen. 2. Manometric experiments show that α-glycerophosphate enhances the respiration of lens preparations. This respiration can be further increased by the addition of ADP and is abolished by cyanide and antimycin. The inference from these experiments is that a mitochondrial system able to oxidize α-glycerophosphate is present, i.e. the particulate half of the α-glycerophosphate cycle. 3. More than the calculated proportion of NADH is used when limiting amounts of dihydroxyacetone phosphate are added to lens tissue in spectrophotometric experiments. Dihydroxyacetone phosphate is therefore regenerated and an α-glycerophosphate cycle is operative. 4. A preparation of a particulate α-glycerophosphate dehydrogenase that takes up oxygen with methylene blue as electron acceptor is described. 5. Methods for obtaining mitochondria from lens are compared, and a useful extraction medium is defined. 6. Mitochondria with activities of the same order of magnitude as those obtained from liver, with α-glycerophosphate and glutamate as substrates, are prepared from epithelium detached from the capsule; some respiratory control is observed. PMID:4290550

  11. Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells.

    PubMed

    Maia, Leandro; Dias, Marianne Camargos; de Moraes, Carolina Nogueira; de Paula Freitas-Dell'Aqua, Camila; da Mota, Ligia S L Silveira; Santiloni, Valquíria; da Cruz Landim-Alvarenga, Fernanda

    2017-03-01

    Cryopreservation is a feasible alternative to maintaining several cell lines, particularly for immediate therapeutic use, transportation of samples, and implementation of new in vitro studies. This work parts from the hypothesis that the medium of cryopreservation composed by 90% of conditioned medium (CM) supports cryopreservation of equine umbilical cord intervascular matrix mesenchymal stem cells (UCIM-MSCs), allowing the maintenance of the biological properties for the establishment of cell banks intended for therapeutic use and in vitro studies. Thus, we evaluated the viability, apoptosis/necrosis rates, immunophenotypic profile (IP), chromosomal stability, clonicity, and differentiation potential of UCIM-MSCs cryopreserved with four different mediums (with FBS: M1, M3, M4 and without FBS: M2). After 3 months of cryopreservation, samples were thawed and analyzed. The potential of differentiation in the mesodermal lineages, clonicity, and the chromosomal stability were maintained after cryopreservation of UCIM-MSCs with medium containing FBS. Changes (P < 0.05) at IP for some markers were observed at cells cryopreserved with medium M1-M3. Only the UCIM-MSCs cryopreserved with the CM (M4) had similar viability post-thaw (P = 0.23) when compared with fresh cells. We proved the hypothesis that the medium of cryopreservation containing CM supports the cryopreservation of UCIM-MSCs, at the experimental conditions, being the medium that better maintains the biological characteristics observed at fresh cells. Thus, future studies of UCIM-MSCs secretome should be conducted to better understand the beneficial and protective effects of the CM during the freezing process. © 2017 International Federation for Cell Biology.

  12. [Laboratory simulation study on the influence of resuspension on the release of mercury from Yangtze estuarine tidal flat].

    PubMed

    Li, Meng; Bi, Chun-Juan; Zhang, Jing-Jing; Lü, Jin-Gang; Chen, Zhen-Lou

    2011-11-01

    Using a Particle Environment Simulator, laboratory simulation experiment on the influence of resuspension on the release of mercury from sediments collected from medium tidal flat and low tidal flat in Chongming Dongtan was conducted. Results indicated that the conditions of overlying water changed significantly during resuspension. The concentrations of HgD and HgP were mainly in the rang of 0.152-2.657 microg x L(-1), 0.080-2.722 microg x L(-1) separately. In the resuspension processes, mercury was released from the medium intertidal sediments under the salinity condition of 4.1 per thousand, 8.0 per thousand, 10.0 per thousand, 13.0 per thousand, and from the low intertidal sediments under the salinity condition of 13.0 per thousand. However, the release of mercury from the medium intertidal sediments is not significant under the salinity condition of 4.1 per thousand, 8.0 per thousand, 10.0 per thousand. The salinity of overlying water had an important effect on resuspension. In different salinity condition, the release of mercury was different. And the release of mercury from medium tidal flat and low tidal flat was different during the resuspension periods, the release of mercury from the medium intertidal sediment was significantly higher than the release from the low intertidal sediment. When the rotate speed was (210 +/- 5) r x min(-1), the desorption of mercury significantly increased, thereby the rotate speed had an effect on the release of mercury.

  13. Pericellular oxygen concentration of cultured primary human trophoblasts

    PubMed Central

    Chen, Baosheng; Longtine, Mark S.; Nelson, D. Michael

    2012-01-01

    Introduction Oxygen is pivotal in placental development and function. In vitro culture of human trophoblasts provides a useful model to study this phenomenon, but a hotly debated issue is whether or not the oxygen tension of the culture conditions mimics in vivo conditions. We tested the hypothesis that ambient oxygen tensions in culture reflect the pericellular oxygen levels. Methods We used a microelectrode oxygen sensor to measure the concentration of dissolved oxygen in the culture medium equilibrated with 21%, 8% or <0.5% oxygen. Results The concentration of oxygen in medium without cells resembled that in the ambient atmosphere. The oxygen concentration present in medium bathing trophoblasts was remarkably dependent on the depth within the medium where sampling occurred, and the oxygen concentration within the overlying atmosphere was not reflected in medium immediately adjacent to the cells. Indeed, the pericellular oxygen concentration was in a range that most would consider severe hypoxia, at ≤ 0.6% oxygen or about 4.6 mm Hg, when the overlying atmosphere was 21% oxygen. Conclusions We conclude that culture conditions of 21% oxygen are unable to replicate the pO2 of 40–60 mm Hg commonly attributed to the maternal blood in the intervillous space in the second and third trimesters of pregnancy. We further surmise that oxygen atmospheres in culture conditions between 0.5% and 21% provide different oxygen fluxes in the immediate pericellular environment yet can still yield insights into the responses of human trophoblast to different oxygen conditions. PMID:23211472

  14. Basic FGF Support of Human Embryonic Stem Cell Self-Renewal

    PubMed Central

    Levenstein, Mark E.; Ludwig, Tenneille E.; Xu, Ren-He; Llanas, Rachel A.; VanDenHeuvel-Kramer, Kaitlyn; Manning, Daisy; Thomson, James A.

    2015-01-01

    Human embryonic stem (ES) cells have most commonly been cultured in the presence of basic FGF (FGF2) either on fibroblast feeder layers or in fibroblast-conditioned medium. Recently, it has been reported that elevated concentrations of FGF2 permit the culture of human ES cells in the absence of fibroblasts or fibroblast-conditioned medium. Here we compare the ability of unconditioned medium (UM) supplemented with 4, 24, 40, 80, 100 and 250 ng/ml FGF2 to sustain low-density human ES cell cultures through multiple passages. In these stringent culture conditions, 4, 24, and 40 ng/ml FGF2 failed to sustain human ES cells through three passages, but 100 ng/ml sustained human ES cells with an effectiveness comparable to conditioned medium (CM). Two human ES cell lines (H1 and H9) were maintained for up to 164 population doublings (7 and 4 months) in UM supplemented with 100 ng/ml FGF2. After prolonged culture the cells formed teratomas when injected into SCID-beige mice, and expressed markers characteristic of undifferentiated human ES cells. We also demonstrate that FGF2 is degraded more rapidly in UM than in CM, partly explaining the need for higher concentrations of FGF2 in UM. These results further facilitate the large-scale, routine culture of human ES cells, and suggest that fibroblasts and fibroblast-conditioned medium sustain human ES cells in part by stabilizing FGF signaling above a critical threshold. PMID:16282444

  15. Spray-dried chitosan as a direct compression tableting excipient.

    PubMed

    Chinta, Dakshinamurthy Devanga; Graves, Richard A; Pamujula, Sarala; Praetorius, Natalie; Bostanian, Levon A; Mandal, Tarun K

    2009-01-01

    The objective of this study was to prepare and evaluate a novel spray-dried tableting excipient using a mixture of chitosan and lactose. Three different grades of chitosan (low-, medium-, and high-molecular-weight) were used for this study. Propranolol hydrochloride was used as a model drug. A specific amount of chitosan (1, 1.9, and 2.5 g, respectively) was dissolved in 50 mL of an aqueous solution of citric acid (1%) and later mixed with 50 mL of an aqueous solution containing lactose (20, 19.1, and 18.5 g, respectively) and propanolol (2.2 g). The resultant solution was sprayed through a laboratory spray drier at 1.4 mL/min. The granules were evaluated for bulk density, tap density, Carr index, particle size distribution, surface morphology, thermal properties, and tableting properties. Bulk density of the granules decreased from 0.16 to 0.13 g/mL when the granules were prepared using medium- or high-molecular-weight chitosan compared with the low-molecular-weight chitosan. The relative proportion of chitosan also showed a significant effect on the bulk density. The granules prepared with 1 g of low-molecular-weight chitosan showed the minimum Carr index (11.1%) indicating the best flow properties among all five formulations. All three granules prepared with 1 g chitosan, irrespective of their molecular weight, showed excellent flow properties. Floating tablets prepared by direct compression of these granules with sodium bicarbonate showed 50% drug release between 30 and 35 min. In conclusion, the spray-dried granules prepared with chitosan and lactose showed excellent flow properties and were suitable for tableting.

  16. System and method of infrared matrix-assisted laser desorption/ionization mass spectrometry in polyacrylamide gels

    DOEpatents

    Haglund, Jr., Richard F.; Ermer, David R.; Baltz-Knorr, Michelle Lee

    2004-11-30

    A system and method for desorption and ionization of analytes in an ablation medium. In one embodiment, the method includes the steps of preparing a sample having analytes in a medium including at least one component, freezing the sample at a sufficiently low temperature so that at least part of the sample has a phase transition, and irradiating the frozen sample with short-pulse radiation to cause medium ablation and desorption and ionization of the analytes. The method further includes the steps of selecting a resonant vibrational mode of at least one component of the medium and selecting an energy source tuned to emit radiation substantially at the wavelength of the selected resonant vibrational mode. The medium is an electrophoresis medium having polyacrylamide. In one embodiment, the energy source is a laser, where the laser can be a free electron laser tunable to generate short-pulse radiation. Alternatively, the laser can be a solid state laser tunable to generate short-pulse radiation. The laser can emit light at various ranges of wavelength.

  17. 20 CFR 220.131 - Work which exists in the national economy.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... and medium jobs exist in the national economy: (1) Dictionary of Occupational Titles, published by the..., also published by the Bureau of the Census. (4) Occupational Analyses, prepared for the Social Security...

  18. 20 CFR 220.131 - Work which exists in the national economy.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... and medium jobs exist in the national economy: (1) Dictionary of Occupational Titles, published by the..., also published by the Bureau of the Census. (4) Occupational Analyses, prepared for the Social Security...

  19. 20 CFR 220.131 - Work which exists in the national economy.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... and medium jobs exist in the national economy: (1) Dictionary of Occupational Titles, published by the..., also published by the Bureau of the Census. (4) Occupational Analyses, prepared for the Social Security...

  20. 20 CFR 220.131 - Work which exists in the national economy.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... and medium jobs exist in the national economy: (1) Dictionary of Occupational Titles, published by the..., also published by the Bureau of the Census. (4) Occupational Analyses, prepared for the Social Security...

  1. 20 CFR 220.131 - Work which exists in the national economy.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... and medium jobs exist in the national economy: (1) Dictionary of Occupational Titles, published by the..., also published by the Bureau of the Census. (4) Occupational Analyses, prepared for the Social Security...

  2. Application of pseudo-template molecularly imprinted polymers by atom transfer radical polymerization to the solid-phase extraction of pyrethroids.

    PubMed

    Zhang, Ming; He, Juan; Shen, Yanzheng; He, Weiye; Li, Yuanyuan; Zhao, Dongxin; Zhang, Shusheng

    2018-02-01

    A polymer-based adsorption medium with molecular recognition ability for homologs of pyrethroids was prepared by atom transfer radical polymer iration using a fragment imprinting technique. Phenyl ether-biphenyl eutectic was utilized as a pseudo-template molecule, and the adsorption medium prepared was evaluated by solid-phase extraction and gas chromatography. Selectivity of the medium for pyrethroids was evaluated using it as solid phase extraction packing by Gas Chromatography. The results demonstrated that the absorption amount of bifenthrin, fenpropathrin, permethrin, cypermethrin, fenvalerate, Dursban and pentachloronitrobenzene for molecularly imprinted polymers were 2.32, 2.12, 2.18, 2.20, 2.30, 1.30 and 1.40mgg -1 , respectively, while the non-imprinted polymers were 1.20, 1.13, 1.25, 1.05, 1.20, 1.23 and 1.32mgg -1 , respectively. The rebinding test based on the molecularly imprinted solid phase extraction column technique showed the recoveries of honey sample spiked with seven insecticides within 88.5-106.2%, with relative standard deviations of 2.38-5.63%. Finally, the method was successfully applied to the analysis of pyrethroids in a honey sample. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. The preparedness of hospital Health Information Services for system failures due to internal disasters.

    PubMed

    Lee, Cheens; Robinson, Kerin M; Wendt, Kate; Williamson, Dianne

    The unimpeded functioning of hospital Health Information Services (HIS) is essential for patient care, clinical governance, organisational performance measurement, funding and research. In an investigation of hospital Health Information Services' preparedness for internal disasters, all hospitals in the state of Victoria with the following characteristics were surveyed: they have a Health Information Service/ Department; there is a Manager of the Health Information Service/Department; and their inpatient capacity is greater than 80 beds. Fifty percent of the respondents have experienced an internal disaster within the past decade, the majority affecting the Health Information Service. The most commonly occurring internal disasters were computer system failure and floods. Two-thirds of the hospitals have internal disaster plans; the most frequently occurring scenarios provided for are computer system failure, power failure and fire. More large hospitals have established back-up systems than medium- and small-size hospitals. Fifty-three percent of hospitals have a recovery plan for internal disasters. Hospitals typically self-rate as having a 'medium' level of internal disaster preparedness. Overall, large hospitals are better prepared for internal disasters than medium and small hospitals, and preparation for disruption of computer systems and medical record services is relatively high on their agendas.

  4. Voyager investigation of the cosmic diffuse background: Observations of rocket-studied locations with Voyager

    NASA Technical Reports Server (NTRS)

    Henry, Richard C.

    1994-01-01

    Attachments to this final report include 2 papers connected with the Voyager work: 'Voyager Observations of Dust Scattering Near the Coalsack Nebula' and 'Search for the Intergalactic Medium'. An appendix of 12 one-page write-ups prepared in connection with another program, UVISI, is also included. The one-page write-ups are: (1) Sky survey of UV point sources to 600 times fainter than previous (TD-1) survey; (2) Diffuse galactic light: starlight scattered from dust at high galactic latitude; (3) Optical properties of interstellar grains; (4) Fluorescence of molecular hydrogen in the interstellar medium; (5) Line emission from hot interstellar medium and/or hot halo of galaxy; (6) Integrated light of distant galaxies in the ultraviolet; (7) Intergalactic far-ultraviolet radiation field; (8) Radiation from recombining intergalactic medium; (9) Radiation from re-heating of intergalactic medium following recombination; (10) Radiation from radiative decay of dark matter candidates (neutrino, etc.); (11) Reflectivity of the asteroids in the Ultraviolet; and (12) Zodiacal light.

  5. Enhanced binding by dextran-grafting to Protein A affinity chromatographic media.

    PubMed

    Zhao, Lan; Zhu, Kai; Huang, Yongdong; Li, Qiang; Li, Xiunan; Zhang, Rongyue; Su, Zhiguo; Wang, Qibao; Ma, Guanghui

    2017-04-01

    Dextran-grafted Protein A affinity chromatographic medium was prepared by grafting dextran to agarose-based matrix, followed by epoxy-activation and Protein A coupling site-directed to sulfhydryl groups of cysteine molecules. An enhancement of both the binding performance and the stability was achieved for this dextran-grafted Protein A chromatographic medium. Its dynamic binding capacity was 61 mg immunoglobulin G/mL suction-dried gel, increased by 24% compared with that of the non-grafted medium. The binding capacity of dextran-grafted medium decreased about 7% after 40 cleaning-in-place cycles, much lower than that of the non-grafted medium as decreased about 15%. Confocal laser scanning microscopy results showed that immunoglobulin G was bound to both the outside and the inside of dextran-grafted medium faster than that of non-grafted one. Atomic force microscopy showed that this dextran-grafted Protein A medium had much rougher surface with a vertical coordinate range of ±80 nm, while that of non-grafted one was ±10 nm. Grafted dextran provided a more stereo surface morphology and immunoglobulin G molecules were more easily to be bound. This high-performance dextran-grafted Protein A affinity chromatographic medium has promising applications in large-scale antibody purification. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. A simple and cost effective liquid culture system for the micropropagation of two commercially important apple rootstocks.

    PubMed

    Mehta, Mohina; Ram, Raja; Bhattacharya, Amita

    2014-07-01

    The two commercially important apple rootstocks i.e., MM106 and B9 were micropropagated using a liquid culture system. Three different strengths of 0.8% agar solidified PGR free basal MS medium were first tested to optimize the culture media for both the rootstocks. Full strength medium (MS0) supported maximum in vitro growth, multiplication, rooting and survival under field conditions as opposed to quarter and half strength media. When three different volumes of liquid MS0 were tested, highest in vitro growth, multiplication, rooting and also survival under field conditions were achieved in 20 mL liquid MS0. The cost of one litre of liquid medium was also reduced by 8 times to Rs. 6.29 as compared to solid medium. The cost of 20 mL medium was further reduced to Rs. 0.125.

  7. An improved method for growing neurons: Comparison with standard protocols.

    PubMed

    Pozzi, Diletta; Ban, Jelena; Iseppon, Federico; Torre, Vincent

    2017-03-15

    Since different culturing parameters - such as media composition or cell density - lead to different experimental results, it is important to define the protocol used for neuronal cultures. The vital role of astrocytes in maintaining homeostasis of neurons - both in vivo and in vitro - is well established: the majority of improved culturing conditions for primary dissociated neuronal cultures rely on astrocytes. Our culturing protocol is based on a novel serum-free preparation of astrocyte - conditioned medium (ACM). We compared the proposed ACM culturing method with other two commonly used methods Neurobasal/B27- and FBS- based media. We performed morphometric characterization by immunocytochemistry and functional analysis by calcium imaging for all three culture methods at 1, 7, 14 and 60days in vitro (DIV). ACM-based cultures gave the best results for all tested criteria, i.e. growth cone's size and shape, neuronal outgrowth and branching, network activity and synchronization, maturation and long-term survival. The differences were more pronounced when compared with FBS-based medium. Neurobasal/B27 cultures were comparable to ACM for young cultures (DIV1), but not for culturing times longer than DIV7. ACM-based cultures showed more robust neuronal outgrowth at DIV1. At DIV7 and 60, the activity of neuronal network grown in ACM had a more vigorous spontaneous electrical activity and a higher degree of synchronization. We propose our ACM-based culture protocol as an improved and more suitable method for both short- and long-term neuronal cultures. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Intracerebral Cell Implantation: Preparation and Characterization of Cell Suspensions.

    PubMed

    Rossetti, Tiziana; Nicholls, Francesca; Modo, Michel

    2016-01-01

    Intracerebral cell transplantation is increasingly finding a clinical translation. However, the number of cells surviving after implantation is low (5-10%) compared to the number of cells injected. Although significant efforts have been made with regard to the investigation of apoptosis of cells after implantation, very little optimization of cell preparation and administration has been undertaken. Moreover, there is a general neglect of the biophysical aspects of cell injection. Cell transplantation can only be an efficient therapeutic approach if an optimal transfer of cells from the dish to the brain can be ensured. We therefore focused on the in vitro aspects of cell preparation of a clinical-grade human neural stem cell (NSC) line for intracerebral cell implantation. NSCs were suspended in five different vehicles: phosphate-buffered saline (PBS), Dulbecco's modified Eagle medium (DMEM), artificial cerebral spinal fluid (aCSF), HypoThermosol, and Pluronic. Suspension accuracy, consistency, and cell settling were determined for different cell volume fractions in addition to cell viability, cell membrane damage, and clumping. Maintenance of cells in suspension was evaluated while being stored for 8 h on ice, at room temperature, or physiological normothermia. Significant differences between suspension vehicles and cellular volume fractions were evident. HypoThermosol and Pluronic performed best, with PBS, aCSF, and DMEM exhibiting less consistency, especially in maintaining a suspension and preserving viability under different storage conditions. These results provide the basis to further investigate these preparation parameters during the intracerebral delivery of NSCs to provide an optimized delivery process that can ensure an efficient clinical translation.

  9. Gamma-ray line afterglow from burst environments

    NASA Technical Reports Server (NTRS)

    Fencl, H. S.; Boyd, R. N.; Hartmann, Dieter

    1992-01-01

    We consider photoerosion and direct pair production in a medium surrounding a gamma-ray burst. The resulting secondary gamma-rays may provide diagnostic tools of these environments and, in turn, of the nature of the bursters themselves. In some instances short-lived nuclides are formed; the beta-delayed gamma-rays produced from their decays provide the signatures of the photoerosion. In addition, annihilation radiation produced from positrons resulting from direct pair production is related to the plasma conditions in the medium. We investigate the plausibility of detecting the various radiations. Under extremely favorable conditions, the photoerosion afterglow might be detectable with the present generation of detectors. However, the positron annihilation line should be detectable under a fairly wide range in the conditions of the medium.

  10. Stimulation of plasmin activity in cultured human fibroblast cells by Porphyromonas endodontalis.

    PubMed

    Oikawa, T; Ogura, N; Akiba, M; Abiko, Y; Takiguchi, H; Izumi, H

    1993-09-01

    1. Plasmin activity in the conditioned medium of Gin-1 cells, a human gingival fibroblast cell line, was stimulated by Porphyromonas endodontalis, a putative pathogen of oral submucous abscesses, in a time- and dose-dependent manner. 2. P. endodontalis stimulated the activity of plasminogen activator in both the conditioned medium and the cell lysate. The plasminogen activator in Gin-1 cells was approx. 50 kDa by zymography. 3. The conditioned medium of Gin-1 cells exposed to P. endodontalis stimulated the conversion of human serum prekallikrein to kallikrein. 4. These results suggested that P. endodontalis stimulates the plasminogen activator-plasmin system in Gin-1 cells, and that activated plasmin plays a role in the progress of periodontal tissue inflammation.

  11. Optimization of flask culture medium and conditions for hyaluronic acid production by a Streptococcus equisimilis mutant nc2168.

    PubMed

    Chen, Yong-Hao; Li, Jun; Liu, Li; Liu, Hong-Zhi; Wang, Qiang

    2012-10-01

    A mutant designated NC2168, which was selected from wild-type Streptococcus equisimilis CVCC55116 by ultraviolet ray combined with(60)Co-γ ray treatment and does not produce streptolysin, was employed to produce hyaluronic acid (HA). In order to increase the output of HA in a flask, the culture medium and conditions for NC2168 were optimized in this study. The influence of culture medium ingredients including carbon sources, nitrogen sources and metal ions on HA production was evaluated using factional factorial design. The mathematical model, which represented the effect of each medium component and their interaction on the yield of HA, was established by the quadratic rotary combination design and response surface method. The model estimated that, a maximal yield of HA could be obtained when the concentrations of yeast extract, peptone, glucose, and MgSO4 were set at 3 g/100 mL, 2 g/100 mL, 0.5 g/100 mL and 0.15 g/100 mL, respectively. Compared with the values obtained by other runs in the experimental design, the optimized medium resulted in a remarkable increase in the output of HA and the maximum of the predicted HA production was 174.76 mg/L. The model developed was accurate and reliable for predicting the production of HA by NC2168.Cultivation conditions were optimized by an orthogonal experimental design and the optimal conditions were as follows: temperature 33°C, pH 7.8, agitation speed 200 rpm, medium volume 20 mL.

  12. The effect of environmental pH on polymeric transfection efficiency.

    PubMed

    Kang, Han Chang; Samsonova, Olga; Kang, Sun-Woong; Bae, You Han

    2012-02-01

    Although polymers, polyplexes, and cells are exposed to various extracellular and intracellular pH environments during polyplex preparation and polymeric transfection, the impact of environmental pH on polymeric transfection has not yet been investigated. This study aims to understand the influence of environmental pH on polymeric transfection by modulating the pH of the transfection medium or the culture medium. Changes in the extracellular pH affected polymeric transfection by way of complex factors such as pH-induced changes in polymer characteristics (e.g., proton buffering capacity and ionization), polyplex characteristics (e.g., size, surface charge, and decomplexation), and cellular characteristics (e.g., cellular uptake, cell cycle phases, and intracellular pH environment). Notably, acidic medium delayed endocytosis, endosomal acidification, cytosolic release, and decomplexation of polyplexes, thereby negatively affecting gene expression. However, acidic medium inhibited mitosis and reduced dilution of gene expression, resulting in increased transfection efficiency. Compared to pH 7.4 medium, acidic transfection medium reduced gene expression 1.6-7.7-fold whereas acidic culture medium enhanced transfection efficiency 2.1-2.6-fold. Polymeric transfection was affected more by the culture medium than by the transfection medium. Understanding the effects of extracellular pH during polymeric transfection may stimulate new strategies for determining effective and safe polymeric gene carriers. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Comparison of the root canal debridement ability of two single file systems with a conventional multiple rotary system in long oval-shaped root canals: In vitro study.

    PubMed

    Khoshbin, Elham; Shokri, Abbas; Donyavi, Zakieh; Shahriari, Shahriar; Salehimehr, Golsa; Farhadian, Maryam; Kavandi, Zeinab

    2017-08-01

    This study sought to compare the root canal debridement ability of Neolix, Reciproc and ProTaper rotary systems in long oval-shaped root canals. Eighty five extracted single-rooted human teeth with long oval-shaped single root canals were selected and divided into three experimental groups(n=25) and one control group (n= 10). Root canals were filled with Vitapex radiopaque contrast medium and prepared with Neolix, Reciproc or ProTaper systems. The control group only received irrigation. Digital radiographs were obtained at baseline and postoperatively and subjected to digital subtraction. The percentage of reduction in contrast medium was quantified at 0-5 mm and 5-10 mm distances from the apex. The data were analyzed using one-way ANOVA and t-test. The mean percentage of the contrast medium removed was not significantly different in the 0-5mm segment among the three groups ( P =0.6). In the 5-10mm segment a significant difference was found in this regard among the ProTaper and Reciproc groups ( P =0.02) and the highest mean percentage of contrast medium was removed by ProTaper. But, difference between ProTaper and Neolix as well as Neolix and Reciproc was not significant. In Neolix ( P =0.024) and Reciproc ( P =0.002) systems, the mean percentage of the contrast medium removed from the 0-5mm segment was significantly greater than that in 5-10mm segment; however, this difference was not significant in ProTaper group ( P =0.069). Neolix single-file system may be a suitable alternative to ProTaper multiple-file system in debridement of long oval shaped canals. Key words: Root Canal Preparation, Debridement, Root Canal Therapy.

  14. Geyser preplay and eruption in a laboratory model with a bubble trap

    NASA Astrophysics Data System (ADS)

    Adelstein, Esther; Tran, Aaron; Saez, Carolina Muñoz; Shteinberg, Alexander; Manga, Michael

    2014-09-01

    We present visual observations and temperature measurements from a laboratory model of a geyser. Our model incorporates a bubble trap, a zone in which vapor can accumulate in the geyser's subsurface plumbing, in a vertical conduit connected to a basal chamber. Analogous features have been identified at several natural geysers. We observe three types of eruptions: 1) rising bubbles eject a small volume of liquid in a weak spout (small eruption); 2) boiling occurs in the conduit above the bubble trap (medium eruption); and 3) boiling occurs in the conduit and chamber (large eruption). In the last two cases, boiling in the conduit causes a rapid hydrostatic pressure drop that allows for the rise and eruption of liquid water in a vigorous spout. Boiling initiates at depth rather than propagating downward from the surface. In a single eruption cycle, multiple small eruptions precede every medium and large eruption. At least one eruption cycle that culminates in a medium eruption (i.e., a quiescent period followed by a series of small eruptions leading up to a medium eruption) precedes every eruption cycle that culminates in a large eruption. We find that the transfer of fluid with high enthalpy to the upper conduit during small and medium eruptions is necessary to heat the upper conduit and prepare the system for the full boiling required for a large eruption. The placement of the bubble trap midway up the conduit allows for more efficient heating of the upper conduit. Our model provides insight into the influence of conduit geometry on eruption style and the importance of heat transfer by smaller events in preparing the geyser system for eruption.

  15. Medium components and culture conditions affect the thermotolerance of aerial conidia of fungal biocontrol agent Beauveria bassiana.

    PubMed

    Ying, S-H; Feng, M-G

    2006-09-01

    To produce more thermotolerable conidia of Beauveria bassiana, a well-known fungal biocontrol agent, by optimizing the medium components and culture conditions. The conidia produced on media including 0.5-6% glucose, sucrose or starch as carbon source and 50-300-microg ml(-1) Cu2+, Zn2+, Mn2+ or Fe3+ as additive to Sabouraud dextrose medium at 15-30 degrees C, pH 4-8 or KCl-adjusted water availabilities were exposed to 30-min wet heat stress at 48 degrees C. The medium components for conidial production with greatly enhanced thermotolerance included 4% glucose as optimum or 1% starch as alternative for the carbon source and < or =50-microg ml(-1) Mn2+ for the metal additive. The culture conditions were optimized as 25 degrees C and pH 5-6. Conidial thermotolerance decreased remarkably when sucrose and Fe3+ or Cu2+ were used in the cultures, but altered slightly when 50-200-microg ml(-1) Zn2+ were included. The tolerance of B. bassiana conidia to the thermal stress was significantly affected by the medium composition and culture conditions under which the conidia were produced. Proper treatment of small grains as mass production substrates for more glucose release and supplement of glucose or 50-microg ml(-1) Mn2+ are possible means to enhancing conidial thermotolerance and field persistence for improved insect control.

  16. H2O-EG-assisted synthesis of uniform urchinlike rutile TiO2 with superior lithium storage properties.

    PubMed

    Chen, Jun Song; Liang, Yen Nan; Li, Yongmei; Yan, Qingyu; Hu, Xiao

    2013-10-23

    A facile green method to synthesize uniform nanostructured urchinlike rutile TiO2 is demonstrated. Titanium trichloride was selected as the TiO2 precursor, and a mixed solvent containing H2O and ethylene glycol was used. By using this binary medium, the nucleation and crystal growth of rutile TiO2 can be regulated, giving rise to very uniform urchinlike structures with tailorable sizes. As confirmed by the SEM and TEM analysis, large particles with dense aggregation of needle-like building blocks or small ones with loosely packed subunits could be obtained at different reaction conditions. The as-prepared samples were applied as the anode material for lithium-ion batteries, and they were shown to have superior properties with a high reversible capacity of 140 mA h g(-1) at a high current rate of 10 C for up to 300 cycles, which is almost unmatched by other rutile TiO2-based electrodes. A stable capacity of 88 mA h g(-1) can also be delivered at an extremely high rate of 50 C, suggesting the great potential of the as-prepared product for high-rate lithium-ion batteries.

  17. A Novel Seeding Method of Interfacial Polymerization-Assisted Dip Coating for the Preparation of Zeolite NaA Membranes on Ceramic Hollow Fiber Supports.

    PubMed

    Cao, Yue; Wang, Ming; Xu, Zhen-Liang; Ma, Xiao-Hua; Xue, Shuang-Mei

    2016-09-28

    A novel seeding method combining interfacial polymerization (IP) technique with dip-coating operation was designed for directly coating nanosized NaA seed crystals (150 nm) onto the micrometer-sized α-Al2O3 hollow fiber support, in which the polyamide (PA) produced by IP acted as an effective medium to freeze and fix seed crystals at the proper position so that the controlled seed layer could be accomplished. While a coating suspension with only 0.5 wt % seed content was used, a very thin seed layer with high quality and good adhesion was achieved through dip coating twice without drying between, and the whole seeding process was operated at ambient conditions. The resulting zeolite NaA membranes not only exhibited high pervaporation (PV) performance with an average separation factor above 10000 and flux nearly 9.0 kg/m(2)·h in dehydration of 90 wt % ethanol aqueous solution at 348 K but also demonstrated great reproducibility by testing more than eight batches of zeolite membranes. In addition, this seeding strategy could be readily extended to the preparation of other supported zeolite membranes for a wide range of separation applications.

  18. Improving the industrial production of 6-APA: enzymatic hydrolysis of penicillin G in the presence of organic solvents.

    PubMed

    Abian, Olga; Mateo, César; Fernández-Lorente, Gloria; Guisán, José M; Fernández-Lafuente, Roberto

    2003-01-01

    The hydrolysis of penicillin G in the presence of an organic solvent, used with the purpose of extracting it from the culture medium, may greatly simplify the industrial preparation of 6-APA. However, under these conditions, PGA immobilized onto Eupergit displays very low stability (half-life of 5 h in butanone-saturated water) and a significant degree of inhibition by the organic solvent (30%). The negative effect of the organic solvent strongly depended on the type of solvent utilized: water saturated with butanone (around 28% v/v) had a much more pronounced negative effect than that of methylisobutyl ketone (MIBK) (solubility in water was only 2%). These problems were sorted out by using a new penicillin G acylase derivative designed to work in the presence of organic solvents (with each enzyme molecule surrounded by an hydrophilic artificial environment) and a suitable organic solvent (MIBK). Using such solvent, this derivative kept its activity unaltered for 1 week at 32 degrees C. Moreover, the enzyme activity was hardly inhibited by the presence of the organic solvent. In this way, the new enzyme derivative thus prepared enables simplification of the industrial hydrolysis of penicillin G.

  19. Kinetics of dissolution of UO2 in nitric acid solutions: A multiparametric study of the non-catalysed reaction

    NASA Astrophysics Data System (ADS)

    Cordara, T.; Szenknect, S.; Claparede, L.; Podor, R.; Mesbah, A.; Lavalette, C.; Dacheux, N.

    2017-12-01

    UO2 pellets were prepared by densification of oxides obtained from the conversion of the oxalate precursor. Then characterized in order to perform a multiparametric study of the dissolution in nitric acid medium. In this frame, for each sample, the densification rate, the grain size and the specific surface area of the prepared pellets were determined prior to the final dissolution experiments. By varying the concentration of the nitric acid solution and temperature, three different and successive steps were identified during the dissolution. Under the less aggressive conditions considered, a first transient step corresponding to the dissolution of the most reactive phases was observed at the solid/solution interface. Then, for all the tested conditions, a steady state step was established during which the normalised dissolution rate was found to be constant. It was followed by a third step characterized by a strong and continuous increase of the normalised dissolution rate. The duration of the steady state, also called "induction period", was found to vary drastically as a function of the HNO3 concentration and temperature. However, independently of the conditions, this steady state step stopped at almost similar dissolved material weight loss and dissolved uranium concentration. During the induction period, no important evolution of the topology of the solid/liquid interface was evidenced authorizing the use of the starting reactive specific surface area to evaluate the normalised dissolution rates thus the chemical durability of the sintered pellets. From the multiparametric study of UO2 dissolution proposed, oxidation of U(IV) to U(VI) by nitrate ions at the solid/liquid interface constitutes the limiting step in the overall dissolution mechanism associated to this induction period.

  20. Long-term viability and differentiation of bovine oviductal monolayers: bidimensional versus three-dimensional culture.

    PubMed

    Gualtieri, R; Mollo, V; Braun, S; Barbato, V; Fiorentino, I; Talevi, R

    2012-10-15

    Different in vitro models have been developed to study the interaction of gametes and embryos with the maternal tract. In cattle, the interaction of the oviduct with gametes and embryos have been classically studied using oviductal explants or monolayers (OMs). Explants are well differentiated but have to be used within 24 h after collection, whereas OMs can be used for a longer time after cell confluence but dedifferentiate during culture, losing cell polarity and ciliation. Herein, OMs were cultured either in M199 plus 10% fetal calf serum or in a semidefined culture medium (Gray's medium), in an immersed condition on collagen-coated coated microporous polyester or polycarbonate inserts under air-liquid interface conditions. The influence of culture conditions on long-term viability and differentiation of OMs was evaluated through scanning electron microscopy, localization of centrin and tubulin at the confocal laser scanning microscope, and assessment of maintenance of viability of sperm bound to OMs. Findings demonstrated that OMs cultured in an immersed condition with Gray's medium retain a better morphology, do not exhibit signs of crisis at least until 3 wks postconfluence, and maintain the viability of bound sperm significantly better than parallel OMs cultured in M199 plus 10% fetal calf serum. OM culture with Gray's medium in air-liquid interface conditions on porous inserts promotes cell polarity, ciliation, and maintenance of bound sperm viability at least until 3 wks postconfluence. In conclusion, oviduct culture in Gray's medium in an immersed or air-liquid condition allows long-term culture and, in the latter case, also ciliation of bovine OMs, and may represent in vitro systems that mimick more closely the biological processes modulated by the oviduct in vivo. Copyright © 2012 Elsevier Inc. All rights reserved.

  1. Practical methods for handling human periodontal ligament stem cells in serum-free and serum-containing culture conditions under hypoxia: implications for regenerative medicine.

    PubMed

    Murabayashi, Dai; Mochizuki, Mai; Tamaki, Yuichi; Nakahara, Taka

    2017-07-01

    Stem cell-based therapies depend on the reliable expansion of patient-derived mesenchymal stem cells (MSCs) in vitro. The supplementation of cell culture media with serum is associated with several risks; accordingly, serum-free media are commercially available for cell culture. Furthermore, hypoxia is known to accelerate the expansion of MSCs. The present study aimed to characterize the properties of periodontal ligament-derived MSCs (PDLSCs) cultivated in serum-free and serum-containing media, under hypoxic and normoxic conditions. Cell growth, gene and protein expression, cytodifferentiation potential, genomic stability, cytotoxic response, and in vivo hard tissue generation of PDLSCs were examined. Our findings indicated that cultivation in serum-free medium does not affect the MSC phenotype or chromosomal stability of PDLSCs. PDLSCs expanded in serum-free medium exhibited more active growth than in fetal bovine serum-containing medium. We found that hypoxia does not alter the cell growth of PDLSCs under serum-free conditions, but inhibits their osteogenic and adipogenic cytodifferentiation while enabling maintenance of their multidifferentiation potential regardless of the presence of serum. PDLSCs expanded in serum-free medium were found to retain common MSC characteristics, including the capacity for hard tissue formation in vivo. However, PDLSCs cultured in serum-free culture conditions were more susceptible to damage following exposure to extrinsic cytotoxic stimuli than those cultured in medium supplemented with serum, suggesting that serum-free culture conditions do not exert protective effects against cytotoxicity on PDLSC cultures. The present work provides a comparative evaluation of cell culture in serum-free and serum-containing media, under hypoxic and normoxic conditions, for applications in regenerative medicine.

  2. Production and optimisation of rosmarinic acid by Satureja hortensis L. callus cultures.

    PubMed

    Tepe, Bektas; Sokmen, Atalay

    2007-11-01

    In this study, production and optimisation of rosmarinic acid, a phenolic acid and an economically important metabolite, was investigated in the callus cultures established from the mature seeds of Satureja hortensis L. (summer savory) plant. Gamborg's B5 basal medium, supplemented with indol butyric acid (IBA) (1.00 mg L(-1)), N6-benzyl aminopurine (6-BA) (1.00 mg L(-1)) and sucrose (2.5%, w/v), was employed for the establishment and maintenance of the callus cultures. Applications were individually prepared by preparing the media containing different IBA/6-BA combinations and sucrose concentrations. All of the applications were carried out in the continuous dark. In the applications, where the effects of IBA/6-BA combinations on the growth and rosmarinic acid accumulation were assayed (1-15 applications), the highest biomass yield was obtained from the medium supplemented with 1.00 mg L(-1) IBA and 5.00 mg L(-1) 6-BA. In the case of the rosmarinic acid accumulation, an opposite relationship was determined between the growth and rosmarinic acid production. While the highest biomass yield was obtained from the medium containing 1.00 mg L(-1) IBA and 5.00 mg L(-1) 6-BA, the highest rosmarinic acid accumulation was obtained from the medium supported with 1.00 mg L(-1) IBA and 1.00 mg L(-1) 6-BA. In the applications where the effects of sucrose concentrations on the growth and rosmarinic acid accumulation were examined, the highest biomass yield was obtained from the medium which is supplemented with 5.0% (w/v) sucrose. In this category, the highest rosmarinic acid accumulation was obtained from the medium which is supported with 3.0% (w/v) sucrose. According to the experiments carried out with the wild S. hortensis, it is found to have 25.02+/-1.21 mg g(-1) rosmarinic acid. No differentiation was observed in any callus during the course of this study.

  3. Uptake of gentamicin by separated, viable renal tubules from rabbits.

    PubMed

    Barza, M; Murray, T; Hamburger, R J

    1980-04-01

    The proximal renal tubules have a marked affinity for gentamicin; they also are the major site of nephrotoxicity caused by this drug. The uptake of radiolabeled gentamicin in separated, viable renal tubules prepared by enzymatic digestion of rabbit kidneys was studied. The preparations showed rapid initial uptake of gentamicin followed by continued slower uptake. Accumulation was not affected by pH, but was significantly inhibited by ouabain, dinitrophenol, anoxia, and hypothermia in the absence of evident cellular damage. At gentamicin concentrations of greater than 50 microgram/ml in the medium, there was competition for drug uptake. Gentamicin efflux in tubules that were taken from a medium containing antibiotic and placed into antibiotic-free fluid was slow and incomplete. From these data it appears that gentamicin uptake by separated renal tubules occurs by a process that requires metabolic energy; thereafter, the drug resides in a poorly exchangeable cellular pool.

  4. Determination of dipyrone in pharmaceutical preparations based on the chemiluminescent reaction of the quinolinic hydrazide-H2O2-vanadium(IV) system and flow-injection analysis.

    PubMed

    Pradana Pérez, Juan A; Durand Alegría, Jesús S; Hernando, Pilar Fernández; Sierra, Adolfo Narros

    2012-01-01

    A rapid, economic and sensitive chemiluminescent method involving flow-injection analysis was developed for the determination of dipyrone in pharmaceutical preparations. The method is based on the chemiluminescent reaction between quinolinic hydrazide and hydrogen peroxide in a strongly alkaline medium, in which vanadium(IV) acts as a catalyst. Principal chemical and physical variables involved in the flow-injection system were optimized using a modified simplex method. The variations in the quantum yield observed when dipyrone was present in the reaction medium were used to determine the concentration of this compound. The proposed method requires no preconcentration steps and reliably quantifies dipyrone over the linear range 1-50 µg/mL. In addition, a sample throughput of 85 samples/h is possible. Copyright © 2011 John Wiley & Sons, Ltd.

  5. A study to identify winning strategies for the business community during the next pandemic.

    PubMed

    Spriggs, Martin

    2013-01-01

    This study examines the relationship between the healthcare system and the corporate sector to answer the following research question: how does the healthcare system best prepare small to medium-sized businesses for the next pandemic influenza? Data were collected and collated through a literature review, electronic survey and semi-structured follow-up telephone interviews. The participants were businesses with membership in the Alberta Chambers of Commerce, a provincial lobby group in Alberta, Canada. The findings indicate strategies that were effective in minimising impact to the business community during the H1N1 pandemic and suggest areas for the business community to improve in preparation for the next pandemic influenza. Recommendations focus on establishing new links for communication between the business community and the healthcare sector and improving strategies to increase the resilience of small to medium-sized businesses for the next pandemic influenza.

  6. Coffee husk waste for fermentation production of mosquitocidal bacteria.

    PubMed

    Poopathi, Subbiah; Abidha, S

    2011-12-01

    Coffee husk waste (CHW) discarded as bio-organic waste, from coffee industries, is rich in carbohydrates. The current study emphasizes the management of solid waste from agro-industrial residues for the production of biopesticides (Bacillus sphaericus, and B. thuringiensis subsp. israelensis), to control disease transmitting mosquito vectors. An experimental culture medium was prepared by extracting the filtrates from coffee husk. A conventional culture medium (NYSM) also was prepared. The studies revealed that the quantity of mosquitocidal toxins produced from CHW is at par with NYSM. The bacteria produced in these media, were bioassayed against mosquito vectors (Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti) and it was found that the toxic effect was statistically comparable. Cost-effective analysis have revealed that, production of biopesticides from CHW is highly economical. Therefore, the utilization of CHW provides dual benefits of effective utilization of environmental waste and efficient production of mosquitocidal toxins.

  7. [FEATURES OF MASS-SPECTROMETRIC PROTEIN PROFILES OF STRAINS OF BRUCELLOSIS CAUSATIVE AGENT DURING PREPARATION OF CULTURE ON VARIOUS NUTRIENT MEDIA].

    PubMed

    Ulshina, D V; Kovalev, D A; Zhirov, A M; Zharinova, N V; Khudoleev, A A; Kogotkova, O I; Efremenko, V I; Evchenko, N I; Kulichenko, A N

    2016-01-01

    Carry out comparative analysis using time-of-flight mass-spectrometry with matrix laser desorption/ionization (MALDI-TOF MS) of protein profiles of brucellosis causative agents (Brucella melitensis Rev-1 and Brucella abortus 19BA), cultivated in various nutrient media: Albimi agar, brucellagar and erythrit-agar. Vaccine,strains: Brucella melitensis Rev-1 and Brucella abortus 19BA. Protein profiling in linear mode on Microflex "Bruker Daltonics" MALDI-TOF mass-spectrometer. A number of characteristic features of brucella mass-spectra was detected: in particular, preservation of the total qualitative composition of protein profiles of cultures and significant differences in the intensity of separate peaks depending on the nutrient medium used. Based on the analysis of the data obtained, use of Albimi agar as the nutrient medium for preparation of brucella culture samples for mass-spectrometric analysis was shown to be optimal.

  8. Characterization of brush borders purified in iso-osmotic medium and microvillar membranes subfractionated from mouse small intestine.

    PubMed Central

    Fujita, M; Ohta, H; Uezato, T

    1981-01-01

    Brush borders free of nuclei were isolated by repeated homogenization and centrifugation in iso-osmotic medium. They showed typical morphology under electron microscopy. The mean recovery and enrichment of alkaline phosphatase activity in the brush-border fraction were 50% and 17.5-fold respectively. gamma-Glutamyl transpeptidase showed a close parallelism with alkaline phosphatase and sucrase in subcellular distribution. Microvillar membranes were purified from isolated brush borders; they showed a further enrichment for alkaline phosphatase and were composed of homogeneous vesicles. Both brush-border and microvillar-membrane preparations were analysed for contamination by basolateral and endoplasmic-reticular membranes. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the microvillar-membrane preparation in six different systems revealed approx. 40 components in the mol.wt. range 15 000-232 000. They were grouped into seven major classes on the basis of molecular weight and electrophoretic patterns. Images PLATE 1 PLATE 2 PMID:7317008

  9. Restoration of longitudinal laser tomography target image from inhomogeneous medium degradation under common conditions.

    PubMed

    Yi, WenJun; Wang, Ping; Fu, MeiCheng; Tan, JiChun; Zhu, Jubo; Li, XiuJian

    2017-07-10

    In order to overcome the shortages of the target image restoration method for longitudinal laser tomography using self-calibration, a more general restoration method through backscattering medium images associated with prior parameters is developed for common conditions. The system parameters are extracted from pre-calibration, and the LIDAR ratio is estimated according to the medium types. Assisted by these prior parameters, the degradation caused by inhomogeneous turbid media can be established with the backscattering medium images, which can further be used for removal of the interferences of turbid media. The results of simulations and experiments demonstrate that the proposed image restoration method can effectively eliminate the inhomogeneous interferences of turbid media and achieve exactly the reflectivity distribution of targets behind inhomogeneous turbid media. Furthermore, the restoration method can work beyond the limitation of the previous method that only works well under the conditions of localized turbid attenuations and some types of targets with fairly uniform reflectivity distributions.

  10. Improving the corrosion wear resistance of AISI 316L stainless steel by particulate reinforced Ni matrix composite alloying layer

    NASA Astrophysics Data System (ADS)

    Xu, Jiang; Zhuo, Chengzhi; Tao, Jie; Jiang, Shuyun; Liu, Linlin

    2009-01-01

    In order to overcome the problem of corrosion wear of AISI 316L stainless steel (SS), two kinds of composite alloying layers were prepared by a duplex treatment, consisting of Ni/nano-SiC and Ni/nano-SiO2 predeposited by brush plating, respectively, and subsequent surface alloying with Ni-Cr-Mo-Cu by a double glow process. The microstructure of the two kinds of nanoparticle reinforced Ni-based composite alloying layers was investigated by means of SEM and TEM. The electrochemical corrosion behaviour of composite alloying layers compared with the Ni-based alloying layer and 316L SS under different conditions was characterized by potentiodynamic polarization test and electrochemical impedance spectroscopy. Results showed that under alloying temperature (1000 °C) conditions, amorphous nano-SiO2 particles still retained the amorphous structure, whereas nano-SiC particles were decomposed and Ni, Cr reacted with SiC to form Cr6.5Ni2.5Si and Cr23C6. In static acidic solution, the corrosion resistance of the composite alloying layer with the brush plating Ni/nano-SiO2 particles interlayer is lower than that of the Ni-based alloying layer. However, the corrosion resistance of the composite alloying layer with the brush plating Ni/nano-SiO2 particles interlayer is prominently superior to that of the Ni-based alloying layer under acidic flow medium condition and acidic slurry flow condition. The corrosion resistance of the composite alloying layer with the brush plating Ni/nano-SiC particles interlayer is evidently lower than that of the Ni-based alloying layer, but higher than that of 316L SS under all test conditions. The results show that the highly dispersive nano-SiO2 particles are helpful in improving the corrosion wear resistance of the Ni-based alloying layer, whereas carbides and silicide phase are deleterious to that of the Ni-based alloying layer due to the fact that the preferential removal of the matrix around the precipitated phase takes place by the chemical attack of an aggressive medium.

  11. Interaction effect of response medium and working memory capacity on creative idea generation

    PubMed Central

    Hao, Ning; Yuan, Huan; Cheng, Rui; Wang, Qing; Runco, Mark A.

    2015-01-01

    This study aimed to examine the interaction effect of response medium (i.e., write down ideas and orally report ideas) and working memory capacity (WMC) on creative idea generation. Participants (N = 90) with higher or lower WMC were asked to solve Alternative Uses Task (AUT) problems in the condition of writing down or speaking out ideas. The results showed that fluency of AUT performance was higher in the writing than in the speaking condition. Additionally, participants with higher WMC performed better on AUT fluency than those with lower WMC in the writing condition, while they showed no difference in the speaking condition. Moreover, level of cognitive demand fully mediated the effect of response medium on AUT fluency. Theoretically, these findings indicated the importance of WMC in creative idea generation, which supported the controlled-attention theory of creativity. Practical implications and future directions were discussed. PMID:26528227

  12. Interaction effect of response medium and working memory capacity on creative idea generation.

    PubMed

    Hao, Ning; Yuan, Huan; Cheng, Rui; Wang, Qing; Runco, Mark A

    2015-01-01

    This study aimed to examine the interaction effect of response medium (i.e., write down ideas and orally report ideas) and working memory capacity (WMC) on creative idea generation. Participants (N = 90) with higher or lower WMC were asked to solve Alternative Uses Task (AUT) problems in the condition of writing down or speaking out ideas. The results showed that fluency of AUT performance was higher in the writing than in the speaking condition. Additionally, participants with higher WMC performed better on AUT fluency than those with lower WMC in the writing condition, while they showed no difference in the speaking condition. Moreover, level of cognitive demand fully mediated the effect of response medium on AUT fluency. Theoretically, these findings indicated the importance of WMC in creative idea generation, which supported the controlled-attention theory of creativity. Practical implications and future directions were discussed.

  13. Effect of cultivating conditions on alpha-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation.

    PubMed

    Liu, Cai-qin; Chen, Qi-he; Cheng, Qian-jun; Wang, Jin-ling; He, Guo-qing

    2007-05-01

    The work is intended to achieve optimum culture conditions of alpha-galactosidase production by a mutant strain Aspergillus foetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of alpha-galactosidase production in SSF were 60% initial moisture of medium, 28 degrees C incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0 approximately 6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum alpha-galactosidase production was 2 037.51 U/g dry matter near the 144th hour of fermentation.

  14. Effect of cultivating conditions on α-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation

    PubMed Central

    Liu, Cai-qin; Chen, Qi-he; Cheng, Qian-jun; Wang, Jin-ling; He, Guo-qing

    2007-01-01

    The work is intended to achieve optimum culture conditions of α-galactosidase production by a mutant strain Aspergillus foetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of α-galactosidase production in SSF were 60% initial moisture of medium, 28 °C incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0~6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum α-galactosidase production was 2 037.51 U/g dry matter near the 144th hour of fermentation. PMID:17542067

  15. Novel microfluidic device for the continuous separation of cancer cells using dielectrophoresis.

    PubMed

    Alazzam, Anas; Mathew, Bobby; Alhammadi, Falah

    2017-03-01

    We describe the design, microfabrication, and testing of a microfluidic device for the separation of cancer cells based on dielectrophoresis. Cancer cells, specifically green fluorescent protein-labeled MDA-MB-231, are successfully separated from a heterogeneous mixture of the same and normal blood cells. MDA-MB-231 cancer cells are separated with an accuracy that enables precise detection and counting of circulating tumor cells present among normal blood cells. The separation is performed using a set of planar interdigitated transducer electrodes that are deposited on the surface of a glass wafer and slightly protrude into the separation microchannel at one side. The device includes two parts, namely, a glass wafer and polydimethylsiloxane element. The device is fabricated using standard microfabrication techniques. All experiments are conducted with low conductivity sucrose-dextrose isotonic medium. The variation in response between MDA-MB-231 cancer cells and normal cells to a certain band of alternating-current frequencies is used for continuous separation of cells. The fabrication of the microfluidic device, preparation of cells and medium, and flow conditions are detailed. The proposed microdevice can be used to detect and separate malignant cells from heterogeneous mixture of cells for the purpose of early screening for cancer. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Cellulose nanofiber aerogel as a promising biomaterial for customized oral drug delivery

    PubMed Central

    Bhandari, Jyoti; Mishra, Harshita; Mishra, Pawan Kumar; Wimmer, Rupert; Ahmad, Farhan J; Talegaonkar, Sushama

    2017-01-01

    Cellulose nanofiber (CNF) aerogels with favorable floatability and mucoadhesive properties prepared by the freeze-drying method have been introduced as new possible carriers for oral controlled drug delivery system. Bendamustine hydrochloride is considered as the model drug. Drug loading was carried out by the physical adsorption method, and optimization of drug-loaded formulation was done using central composite design. A very lightweight-aerogel-with-matrix system was produced with drug loading of 18.98%±1.57%. The produced aerogel was characterized for morphology, tensile strength, swelling tendency in media with different pH values, floating behavior, mucoadhesive detachment force and drug release profiles under different pH conditions. The results showed that the type of matrix was porous and woven with excellent mechanical properties. The drug release was assessed by dialysis, which was fitted with suitable mathematical models. Approximately 69.205%±2.5% of the drug was released in 24 hours in medium of pH 1.2, whereas ~78%±2.28% of drug was released in medium of pH 7.4, with floating behavior for ~7.5 hours. The results of in vivo study showed a 3.25-fold increase in bioavailability. Thus, we concluded that CNF aerogels offer a great possibility for a gastroretentive drug delivery system with improved bioavailability. PMID:28352172

  17. Optical coatings on laser crystals for HiPER project

    NASA Astrophysics Data System (ADS)

    Oulehla, Jindrich; Pokorný, Pavel; Lazar, Josef

    2011-12-01

    In this contribution we present a technology for deposition of interference coatings for optical components designed to operate as active media in power pulsed lasers. The aim of the technology is to prepare crystals for lasers for the HiPER project (High Power laser Energy Research facility) which should demonstrate the feasibility of laser driven fusion as a future energy source. Diode pumped solid state lasers (DPSSL) are the most likely option for fusion ignition. The choice of the material for the lasers' active medium is critical. Some of the most important properties include the ability to be antireflection coated to reduce the energy losses and increase the overall efficiency. This contribution deals with some of the materials considered to be candidates for slabs serving as the active medium of the DPSSLs. We tested Yb:YAG and Yb:CaF2 samples. As large amounts of heat need to be dissipated during laser operation, cryogenic cooling is necessary. Appropriate coating materials and techniques need to be chosen. Therefore differences between available coating techniques are investigated in terms of adhesion, enduring of stress from temperature shocks, etc. Coated samples were placed into cryogenic environment in order to simulate conditions similar to those in real life operation. Optical microscopy was used for coating investigation after the conducted experiments.

  18. AR coatings on laser crystals for HiPER project

    NASA Astrophysics Data System (ADS)

    Oulehla, Jindřich; Pokorný, Pavel

    2010-08-01

    In this contribution we present a technology for deposition of interference coatings for optical components designed to operate as active media in power pulsed lasers. The aim of the technology is to prepare crystals for lasers for the HiPER project (High Power laser Energy Research) which should demonstrate the feasibility of laser driven fusion as a future energy source. Diode pumped solid state lasers (DPSSL) are the most likely option for fusion ignition. The choice of material for the lasers active medium is critical. Some of the most important properties include the ability to be antireflection coated to reduce the energy losses and increase the overall efficiency. This contribution deals with some of the materials considered to be candidates for slabs serving as the active medium of the DPSSLs. We tested Yb:YAG, Yb:CaF2 samples. As large amounts of heat need to be dissipated during laser operation, cryogenic cooling is necessary. Appropriate coating materials and techniques need to be chosen. Therefore differences between available coating techniques are investigated in terms of adhesion, enduring of stress resulting from temperature shocks, etc. Coated samples were placed into cryogenic environment in order to simulate conditions similar to those in real life operation. Optical microscopy was used for coating investigation after the conducted experiments.

  19. Tripartite entanglement dynamics and entropic squeezing of a three-level atom interacting with a bimodal cavity field

    NASA Astrophysics Data System (ADS)

    Faghihi, M. J.; Tavassoly, M. K.; Bagheri Harouni, M.

    2014-04-01

    In this paper, we study the interaction between a Λ-type three-level atom and two quantized electromagnetic fields which are simultaneously injected in a bichromatic cavity surrounded by a Kerr medium in the presence of field-field interaction (parametric down conversion) and detuning parameters. By applying a canonical transformation, the introduced model is reduced to a well-known form of the generalized Jaynes-Cummings model. Under particular initial conditions which may be prepared for the atom and the field, the time evolution of the state vector of the entire system is analytically evaluated. Then, the dynamics of the atom is studied through the evolution of the atomic population inversion. In addition, two different measures of entanglement between the tripartite system (three entities make the system: two field modes and one atom), i.e., von Neumann and linear entropy are investigated. Also, two kinds of entropic uncertainty relations, from which entropy squeezing can be obtained, are discussed. In each case, the influences of the detuning parameters and Kerr medium on the above nonclassicality features are analyzed in detail via numerical results. It is illustrated that the amount of the above-mentioned physical phenomena can be tuned by choosing the evolved parameters, appropriately.

  20. Improved human islet preparations using Glucocorticoid and Exendin-4

    PubMed Central

    Miki, Atsushi.; Ricordi, Camillo.; Yamamoto, Toshiyuki.; Sakuma, Yasunaru.; Misawa, Ryosuke.; Mita, Atsuyoshi.; Inverardi, Luca.; Alejandro, Rodolfo; Ichii, Hirohito.

    2014-01-01

    Objectives The effects of Glucocorticoid during culture on human islet cells have been controversial. Exendin-4 (EX) enhances the insulin secretion and significantly improves clinical outcomes in islet cell transplantation. In this study, we examined the effects of Glucocorticoids and exendin-4 on human islet cells during pre-transplant culture. Methods Methylprednisolone (MP) and/or EX were added to the standard culture medium for clinical islet cell transplantation. Islets were cultured for 24 hours with three different conditions (Control: no additives, MP alone, MP+EX). Beta cell fractional viability, cellular composition, multiple cytokine/chemokine production, multiple phosphorylation proteins and glucose induced insulin secretion were evaluated. Results Viable beta cell survival in MP and MP+EX group was significantly higher than in the control group. EX prevented MP induced reduction of insulin secretion. MP supplementation to the culture medium decreased cytokine and chemokine production. Moreover, Erk1/2 phosphorylation was significantly increased by MP and MP+EX. Conclusions Glucocorticoid supplementation into culture media significantly decreased the cytokine/chemokine production and increased the Erk1/2 phosphorylation, resulting in the improvement of human beta cell survival. In addition, EX maintained the insulin secretion suppressed by MP. The supplementation of MP and EX together could be a useful strategy to create suitable human islets for transplantation. PMID:25036907

  1. Xanthan production on polyurethane foam and its enhancement by air pressure pulsation.

    PubMed

    Zhang, Zhi-guo; Chen, Hong-zhang

    2010-12-01

    In this study, we evaluated the feasibility of solid-state fermentation (SSF) on polyurethane foam (PUF) for xanthan production. The effects of air pressure pulsation (APP) on biomass accumulation and final xanthan concentration were also studied. Under suitable conditions (15% inoculum, 0.5-cm (side length) PUF cubes, 15 mL medium per gram cubes and 4.5 cm bed depth), the broth was dispersed on the PUF as a film. When the initial glucose concentration in the media was low (20 and 40 g L⁻¹), there was no significant difference between the final xanthan concentration in static SSF and submerged fermentation (SMF). When high initial glucose concentrations (60 and 80 g L⁻¹) were used, the final gum concentrations in SSF were much higher than those in SMF. When the APP technique was applied in xanthan production with a medium containing a high glucose concentration (80 g L⁻¹), the oxygen consumption rate of Xanthomonas campestris was significantly enhanced at the later stages of fermentation, and both the biomass and xanthan concentration were improved. The results indicated that SSF on PUF is suitable for xanthan preparation, especially when the initial glucose concentration ranged from 60 to 80 g L⁻¹. Those results also demonstrated that APP technology can be used to enhance xanthan yields.

  2. Effect of residual monomer from polyacrylamide on head lettuce grown in peat substrate.

    PubMed

    Mroczek, Ewelina; Kleiber, Tomasz; Konieczny, Piotr; Waśkiewicz, Agnieszka

    2015-01-01

    The paper investigates the migration of the acrylamide monomer (AMD) to lettuce chosen as a test plant growing in an organic medium (peat substrate). Polyacrylamide (PAM)-based flocculant added to the growing medium contained no more than 1000 mg kg(-1) of AMD. Plants were grown with varied doses of PAM preparation (0.5-3.0 mg dm(-3) of peat substrate) to compare the results with the control sample. The determination of AMD content, chlorophyll content, weight of the lettuce head, and also analysis of macro- and micro-elements in lyophilised test material was made under the same analytical conditions. The results showed that lettuce plants absorb AMD to the leaves from the peat substrate. The AMD uptake has a negative impact on the growth of lettuce. It reduces the average fresh weight of heads and destabilises the mineral composition of the plant. Therefore, concern related to the transfer risk of the residual AMD from sludge used for organic fertilisation of edible plants still remains a crucial question from a food and consumer safety point of view. To ensure consumer safety, the fate of the AMD following the application of PAM to cropland should be carefully monitored in the whole food chain.

  3. Cost analysis in laccase production.

    PubMed

    Osma, Johann F; Toca-Herrera, José L; Rodríguez-Couto, Susana

    2011-11-01

    In this paper the cost of producing the enzyme laccase by the white-rot fungus Trametes pubescens under both submerged (SmF) and solid-state fermentation (SSF) conditions was studied. The fungus was cultured using more than 45 culture medium compositions. The cost of production was estimated by analyzing the cost of the culture medium, the cost of equipment and the operating costs. The cost of the culture medium represented, in all cases, the highest contribution to the total cost, while, the cost of equipment was significantly low, representing less than 2% of the total costs. The cultivation under SSF conditions presented a final cost 50-fold lower than the one obtained when culturing under SmF conditions at flask scale. In addition, the laccase production under SSF conditions in tray bioreactors reduced the final cost 4-fold compared to the one obtained under SSF conditions at flask scale, obtaining a final price of 0.04 cent €/U. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. Fish meal extract bile esculin agar (FMBE) a selective medium for Bacteroides fragilis group.

    PubMed

    Beena, V K; Rao, S; Kotian, M; Shivananda, P G

    1997-07-01

    Fish meal extract bile esculin agar (FMBE) is prepared using Fish meal extract concentrate as the basal substance, for the selective isolation and presumptive identification of B.fragilis group. The efficiency of the medium was evaluated by growing stock cultures of B.fragilis groups as well as inoculating clinical specimens and comparing the results with Bacteroides bile esculin agar (BBE). All the 87 stock cultures of B.fragilis grew on FMBE and BBE. No other anaerobes tested grew on the medium. However 7 out of 65 neomycin resistant aerobes grew on the FMBE. From the 100 clinical samples, 62 strains of B. Fragilis group were recovered on FMBE and BBE, and 53 strains on supplemented BHIBA. The cost effectiveness, selectivity and the ability to detect esculin hydrolysis will enable FMBE as a suitable medium as comparable to that of BBE, if not superior.

  5. Medium-chain fatty acid synthesis in lactating-rabbit mammary gland. Intracellular concentration and specificity of medium-chain acyl thioester hydrolase.

    PubMed Central

    Knudsen, J

    1979-01-01

    The concentration of medium-chain acyl thioester hydrolase and of fatty acid synthetase was determined by rocket immunoelectrophoresis in nine different particle-free supernatant fractions from lactating-rabbit mammary gland. The molar ratio of the hydrolase to fatty acid synthetase was 1.99 +/- 0.66 (mean +/- S.D.). A rate-limiting concentration of malonyl-CoA was required to ensure the predominant synthesis of medium-chain fatty acids when 2 mol of the hydrolase was added per mol of fatty acid synthetase. The interaction of the hydrolase with fatty acid synthetase was concentration-dependent, though an optimum concentration of hydrolase to synthetase could not be obtained. The lactating-rabbit mammary gland hydrolase altered the pattern of fatty acids synthesized by fatty acid synthetases prepared from cow, goat, sheep and rabbit lactating mammary glands, rabbit liver and cow adipose tissue. PMID:574008

  6. The growth of Treponema hyodysenteriae and other porcine intestinal spirochaetes in a liquid medium.

    PubMed

    Lemcke, R M; Bew, J; Burrows, M R; Lysons, R J

    1979-05-01

    A new simple method for the preparation of a liquid medium containing rabbit serum for the propagation of Treponema hyodysenteriae and other porcine intestinal spirochaetes is described. The medium, when dispensed in shallow layers and sealed under 10 per cent CO2 in nitrogen, had a redox potential not greater than -125mV and an initial pH of about 6.9 when buffered with bicarbonate. Growth of T hyodysenteriae developed more rapidly and viable counts reached higher levels at 42 degrees C than at 37 degrees C. Viable counts increased at least 10,000-fold after two to five days' incubation, depending on the temperature. Growth could be initiated from small inocula that failed to produce colonies on blood agar. Using a 1 per cent inoculum, the medium supported the growth of two strains of T hyodysenteriae through 10 serial passages.

  7. [Experimental study on co-culture of salivary adenoid cystic carcinoma cells and ganglia].

    PubMed

    Gu, Ling; Bu, Rong-fa; Wang, Dong-sheng; E, Ling-ling; Zhu, Guo-xiong

    2012-01-01

    To construct the co-culture models of salivarya denoid cystic carcinoma (SACC) cells and dorsal root ganglia (DRG) of chickens and investigate the promotive effects of SACC on neural tissue. Glass-base culture dish was adopted to construct co-culture model of SACC-83 cells and DRG. SACC-83 cells were seeded in the medium pore with DRG around them. Outgrowth of neuronal processes was observed. Then DRG was cultured in the conditioned medium of SACC-83, with the groups of conditioned medium of MC3T3-E1 and HGF, the group of cell lysis buffer, the groups of serum-free medium and serum-plus medium as the controls. Outgrowth of neuronal processes was also recorded and compared with control groups. In the co-culture model of tumor and neuronal tissue, SACC-83 cells produced a suitable microenvironment in which neuronal processes remarkably grow. Neuronal processes of most DRG displayed growth tendency toward SACC. The group of conditioned medium from SACC-83 manifested obvious promotive effects on DRG. Co-culture model of tumor and neuronal tissue was successfully constructed, with which the promotive effects of tumor on outgrowth of neuronal processes could be observed. So hypothesized that SACC could secrete some neurotrophic factors to guide peripheral nerves gemmating and to trigger the cascade of the neural invasion in succession.

  8. p-Coumaric Acid Attenuates UVB-Induced Release of Stratifin from Keratinocytes and Indirectly Regulates Matrix Metalloproteinase 1 Release from Fibroblasts

    PubMed Central

    Seok, Jin Kyung

    2015-01-01

    Ultraviolet (UV) radiation-induced loss of dermal extracellular matrix is associated with skin photoaging. Recent studies demonstrated that keratinocyte-releasable stratifin (SFN) plays a critical role in skin collagen metabolism by inducing matrix metalloproteinase 1 (MMP1) expression in target fibroblasts. In the present study, we examined whether SFN released from UVB-irradiated epidermal keratinocytes increases MMP1 release from dermal fibroblasts, and whether these events are affected by p-coumaric acid (p-CA), a natural phenolic compound with UVB-shielding and antioxidant properties. HaCaT cells were exposed to UVB in the absence and presence of p-CA, and the conditioned medium was used to stimulate fibroblasts in medium transfer experiments. The cells and media were analyzed to determine the expressions/releases of SFN and MMP1. UVB exposure increased SFN release from keratinocytes into the medium. The conditioned medium of UVB-irradiated keratinocytes increased MMP1 release from fibroblasts. The depletion of SFN using a siRNA rendered the conditioned medium of UVB-irradiated keratinocytes ineffective at stimulating fibroblasts to release MMP1. p-CA mitigated UVB-induced SFN expression in keratinocytes, and attenuated the MMP1 release by fibroblasts in medium transfer experiments. In conclusion, the present study demonstrated that the use of UV absorbers such as p-CA would reduce UV-induced SFN-centered signaling events involved in skin photoaging. PMID:25954129

  9. p-Coumaric Acid Attenuates UVB-Induced Release of Stratifin from Keratinocytes and Indirectly Regulates Matrix Metalloproteinase 1 Release from Fibroblasts.

    PubMed

    Seok, Jin Kyung; Boo, Yong Chool

    2015-05-01

    Ultraviolet (UV) radiation-induced loss of dermal extracellular matrix is associated with skin photoaging. Recent studies demonstrated that keratinocyte-releasable stratifin (SFN) plays a critical role in skin collagen metabolism by inducing matrix metalloproteinase 1 (MMP1) expression in target fibroblasts. In the present study, we examined whether SFN released from UVB-irradiated epidermal keratinocytes increases MMP1 release from dermal fibroblasts, and whether these events are affected by p-coumaric acid (p-CA), a natural phenolic compound with UVB-shielding and antioxidant properties. HaCaT cells were exposed to UVB in the absence and presence of p-CA, and the conditioned medium was used to stimulate fibroblasts in medium transfer experiments. The cells and media were analyzed to determine the expressions/releases of SFN and MMP1. UVB exposure increased SFN release from keratinocytes into the medium. The conditioned medium of UVB-irradiated keratinocytes increased MMP1 release from fibroblasts. The depletion of SFN using a siRNA rendered the conditioned medium of UVB-irradiated keratinocytes ineffective at stimulating fibroblasts to release MMP1. p-CA mitigated UVB-induced SFN expression in keratinocytes, and attenuated the MMP1 release by fibroblasts in medium transfer experiments. In conclusion, the present study demonstrated that the use of UV absorbers such as p-CA would reduce UV-induced SFN-centered signaling events involved in skin photoaging.

  10. Coconut as a Medium for the Experimental Production of Aflatoxin

    PubMed Central

    Arseculeratne, S. N.; De Silva, L. M.; Wijesundera, S.; Bandunatha, C. H. S. R.

    1969-01-01

    Fresh, grated coconut has been found to be an excellent medium for aflatoxin production by Aspergillus flavus. Under optimal conditions, yields of 8 mg of total aflatoxin per g of substrate were obtained. Continuous agitation of the growth medium under moist conditions at 24 C produced highest yields. Aflatoxin was assayed both biologically and chromatographically. The aflatoxin content of cultures varied biphasically with the duration of incubation. It is suggested that this pattern could result from the sequential operation of factors promoting aflatoxin formation on the one hand and a detoxifying mechanism on the other. Images PMID:5803632

  11. Effect of timing and method of post space preparation on sealing ability of remaining root filling material: in vitro microbiological study.

    PubMed

    Grecca, Fabiana Soares; Rosa, Angela Rezende Gomes; Gomes, Maximiliano Schünke; Parolo, Clarissa Fatturi; Bemfica, Jules Renan Dutra; Frasca, Luis Carlos da Fontoura; Maltz, Marisa

    2009-10-01

    To evaluate the effect of timing (immediate versus delayed) and technique of post space preparation on the ability of the residual root canal obturation to prevent coronal bacterial leakage. Sixty-six single-rooted teeth were decoronated at the cementoenamel junction. The canals were prepared according to a step-back technique and were filled with thermoplasticized gutta-percha and AH Plus endodontic sealer (Dentsply De Trey). The root segments were randomly assigned to 8 groups. The positive controls (n = 3) were instrumented but not obturated. The negative controls (n = 3) were instrumented, obturated and sealed with Cavit (3M ESPE). In the other 6 groups (n = 10 each), the post space was prepared either immediately after obturation or 7 days later using LA Axxess burs (SybronEndo) (groups 1 and 2), heated pluggers (groups 3 and 4) or solvent delivered with a hand file (groups 5 and 6). The external surface of all roots was rendered waterproof with nail varnish. Custom-made dual-chamber devices were used to evaluate leakage. The coronal third of the prepared root canal was kept in contact with artificial saliva contaminated with Enterococcus faecalis, and the root apex was submerged in tryptic soy agar medium. The root assemblies were stored at 37 degrees C and were monitored daily over a 90-day period. The occurrence of turbidity in the medium was deemed to indicate bacterial leakage, from which failure of the seal was inferred. Throughout the experimental period, there was no significant difference (p = 0.094) among the preparation techniques, either immediate or delayed, in terms of bacterial leakage. Immediate and delayed post space preparation yielded similar outcomes in terms of the canal seal. Regardless of the timing and the technique of post space preparation, coronal bacterial leakage occurred over time.

  12. Cytotoxic and hemolytic effects of Tritrichomonas foetus on mammalian cells.

    PubMed Central

    Burgess, D E; Knoblock, K F; Daugherty, T; Robertson, N P

    1990-01-01

    Geographically distinct lines of Tritrichomonas foetus were assayed for their ability to cause cytotoxicity in nucleated mammalian cells and lysis of bovine erythrocytes. T. foetus was highly cytotoxic toward a human cervical cell line (HeLa) and early bovine lymphosarcoma (BL-3) but displayed low levels of cytotoxicity against African green monkey kidney (Vero) cells. In addition to variation in the extent of cytotoxicity toward different targets, differences in the levels of cytotoxicity in the same nucleated target occurred with different parasite lines. Whole T. foetus, unfractionated whole-cell extracts, and parasite-conditioned medium (RPMI 1640 without serum) all caused lysis of bovine erythrocytes. Lytic activity in the conditioned medium was substantially reduced by repeated freezing and thawing or heating to 90 degrees C for 30 min. Damage of mammalian target cells by live T. foetus could be reduced by the presence of protease inhibitors; however, such inhibitors did not diminish the lytic effects of conditioned medium. These results suggested that proteolytic enzymes were necessary for the lytic mechanism of the live parasites but were not required once lytic factors were released into the parasite-conditioned medium. They further suggested that the lytic molecules were either proteins or had proteinaceous components. Images PMID:2228233

  13. Race Does Not Predict Melanocyte Heterogeneous Responses to Dermal Fibroblast-Derived Mediators

    PubMed Central

    Sirimahachaiyakul, Pornthep; Sood, Ravi F.; Muffley, Lara A.; Seaton, Max; Lin, Cheng-Ta; Qiao, Liang; Armaly, Jeffrey S.; Hocking, Anne M.; Gibran, Nicole S.

    2015-01-01

    Introduction Abnormal pigmentation following cutaneous injury causes significant patient distress and represents a barrier to recovery. Wound depth and patient characteristics influence scar pigmentation. However, we know little about the pathophysiology leading to hyperpigmentation in healed shallow wounds and hypopigmentation in deep dermal wound scars. We sought to determine whether dermal fibroblast signaling influences melanocyte responses. Methods and Materials Epidermal melanocytes from three Caucasians and three African-Americans were genotyped for single nucleotide polymorphisms (SNPs) across the entire genome. Melanocyte genetic profiles were determined using principal component analysis. We assessed melanocyte phenotype and gene expression in response to dermal fibroblast-conditioned medium and determined potential mesenchymal mediators by proteome profiling the fibroblast-conditioned medium. Results Six melanocyte samples demonstrated significant variability in phenotype and gene expression at baseline and in response to fibroblast-conditioned medium. Genetic profiling for SNPs in receptors for 13 identified soluble fibroblast-secreted mediators demonstrated considerable heterogeneity, potentially explaining the variable melanocyte responses to fibroblast-conditioned medium. Discussion Our data suggest that melanocytes respond to dermal fibroblast-derived mediators independent of keratinocytes and raise the possibility that mesenchymal-epidermal interactions influence skin pigmentation during cutaneous scarring. PMID:26418010

  14. Starch and lipid accumulation in eight strains of six Chlorella species under comparatively high light intensity and aeration culture conditions.

    PubMed

    Takeshita, Tsuyoshi; Ota, Shuhei; Yamazaki, Tomokazu; Hirata, Aiko; Zachleder, Vilém; Kawano, Shigeyuki

    2014-04-01

    The microalgae family Chlorella species are known to accumulate starch and lipids. Although nitrogen or phosphorous deficiencies promote starch and lipids formation in many microalgae, these deficiencies also limit their growth and productivity. Therefore, the Chlorellaceae strains were attempted to increase starch and lipids productivity under high-light-intensity conditions (600-μmol photons m(-2)s(-1)). The 12:12-h light-dark (LD) cycle conditions elicited more stable growth than the continuous light (LL) conditions, whereas the starch and lipids yields increased in LL conditions. The amount of starch and lipids per cell increased in Chlorella viscosa and Chlorella vulgaris in sulfur-deficient medium, and long-chain fatty acids with 20 or more carbon atoms accumulated in cells grown in sulfur-deficient medium. Accumulation of starch and lipids was investigated in eight strains. The accumulation was strain-dependent, and varied according to the medium and light conditions. Five of the eight Chlorella strains exhibited similar accumulation patterns. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. In vitro cultivation of Hysterothylacium aduncum (Nematoda: Anisakidae) from 3rd-stage larvae to egg-laying adults.

    PubMed

    Iglesias, L; Valero, A; Gálvez, L; Benítez, R; Adroher, F J

    2002-11-01

    This is the first demonstration of the in vitro development of the 3rd-stage larvae (L3) of Hysterothylacium aduncum to the adult. This was achieved in a semi-defined medium that is easy to prepare and to reproduce. The L3, collected from the peritoneal cavity of horse mackerel (Trachurus trachurus), were individually inoculated into RPMI-1640 medium +20% heat-inactivated fetal bovine serum (IFBS). It has been demonstrated that the optimum temperature for development is around 13 degrees C and is stimulated by the presence of 5% CO2 in the growth atmosphere, increasing the percentage moulting to the 4th larval stage (L4) by 1.9-fold (from 44 to 82%) and the average survival of the nematodes by 1.6 times (from 60 to 96 days). When the larvae were grown at different pHs, optimum development occurred at pH 4.0. Under these conditions, all the larvae moulted to the L4 and more than two-thirds transformed to the adult stage--in which 25-30% of the females laid eggs--and reached an average survival of over 4 months. When this medium was supplemented with 1% (w/v) of commercial pepsin, all the larvae reached the adult stage, at least 45% of the females oviposited, laying around 12-fold more eggs per female than in the medium without pepsin. The mean size of the eggs (non-fertilized) obtained was 56.8 x 47.6 microm. The mean length of the adult males obtained was between 3.2 and 5.2 cm and the females were between 3.0 and 6.5 cm. The adult specimens were morphologically identified as Hysterothylacium aducum aduncum. This culture medium (RPMI-1640+20% (v/v) IFBS+1 commercial pepsin, at pH 4.0, 13 degrees C and 5% CO2 in air) could facilitate the identification of at least some of the larvae of the genus Hysterothylacium--and perhaps other anisakids--for which the specific identification and the biological study of these parasites is often difficult.

  16. HYDROGENATION OF 4-OXOISOPHORONE OVER PD/AL2/O2 CATALYST UNDER SUPERCRITICAL CO2 MEDIUM

    EPA Science Inventory

    Hydrogenation of 4-oxoisophorone has been studied over 1% Pd/Al2O3 impregnated catalyst in supercritical CO2 medium at different reaction conditions. The effect of temperature, pressure and reaction medium on the conversion and product selectivity is discussed. Phase behavior stu...

  17. Density-dependent regulation of growth of BSC-1 cells in cell culture: growth inhibitors formed by the cells.

    PubMed Central

    Holley, R W; Armour, R; Baldwin, J H

    1978-01-01

    Inhibitors formed by a monkey epithelial cell line, BSC-1, play an important role in limiting growth at high cell densities. At least three inhibitors are formed: lactic acid, ammonia, and an unidentified inhibitor that may be an unstable protein. The unidentified inhibitor is destroyed by shaking the conditioned medium, by bubbling gas through the medium, or by heating or storing the medium in the absence of cells. The concentrations of lactic acid and ammonia that accumulate in conditioned medium inhibit growth when added to fresh medium. These results, together with earlier studies, indicate that density-dependent regulation of growth of BSC-1 cells results from the combined effects of (a) inhibitors formed by the cells, (b) decreased availability of receptor sites for serum growth factors as the cells become crowded, and (c) limiting concentrations of low molecular weight nutrients in the medium. In contrast, density-dependent regulation of growth in 3T3 mouse embryo fibroblasts results almost entirely from inactivation of serum factors. PMID:273914

  18. [Factors inducing transition from growth to dormancy in rhizobacteria Azospirillum brasilense].

    PubMed

    Kushneruk, M A; Tugarova, A V; Il'chukova, A V; Slavkina, E A; Starichkova, N I; Bogatyrev, V A; Antoniuk, L P

    2013-01-01

    The factors suppressing division of the cells of the rhizobacterium Azospirillum brasilense and inducing their transition to a dormant state were analyzed. These included the presence of hexylresorcinol or heavy metals (Cu and Co) in the medium, oxygen stress, and transfer of the cells into the physiological saline or phosphate buffer solution. The results were used to develop a protocol for obtaining of uncultured cells of A. brasilense Sp245, a natural symbiont of wheat. The cells lost their ability to grow on synthetic agar medium, but could revert to growth when incubated in freshly prepared liquid medium. Needle-shaped crystals differing from struvite, which has been previously reported for this strain, were found in the dormant culture of A. brasilense Sp245.

  19. Multiplication in liquid medium of Treponema sp. isolated from intestinal contents of swine.

    PubMed

    Binek, M; Szynkiewicz, Z

    1985-01-01

    Treponema hyodysenteriae and Treponema innocens were multiplied by a simple culture method in liquid medium. TSB medium was prepared by the PRAS method in plasma bottles containing glass beads. Spirochaetes were injected through the rubber stopper and the bottles were incubated while revolving round their axes. The most abundant growth of spirochaetes in rotary culture was observed after 72 h incubation at 40 degrees C. whereas the highest number of viable cells in stationary culture was observed after 120 h. However, in the latter case the number of cells was lower than introduced at inoculation. Growth of the bacteria was stimulated by equine serum and 5% addition of rumen fluid. Optimal growth temperature was 40 degrees C.

  20. Magnetic nanoparticle-supported glutathione: a conceptually sustainable organocatalyst

    EPA Science Inventory

    A conceptually novel nanoparticle-supported and magnetically recoverable organocatalyst has been developed, which is readily prepared from inexpensive starting materials in a truly sustainable manner; which catalyzes Paal-Knorr reaction with high yield in pure aqueous medium that...

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