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Sample records for viability high thermotolerance

  1. High salinity conveys thermotolerance in the coral model Aiptasia

    KAUST Repository

    Gegner, Hagen M.

    2017-12-15

    The endosymbiosis between dinoflagellate algae of the genus Symbiodinium and stony corals provides the foundation of coral reef ecosystems. Coral bleaching, the expulsion of endosymbionts from the coral host tissue as a consequence of heat or light stress, poses a threat to reef ecosystem functioning on a global scale. Hence, a better understanding of the factors contributing to heat stress susceptibility and tolerance is needed. In this regard, some of the most thermotolerant corals also live in particularly saline habitats, but possible effects of high salinity on thermotolerance in corals are anecdotal. Here we test the hypothesis that high salinity may lead to increased thermotolerance. We conducted a heat stress experiment at low, intermediate, and high salinities using a set of host-endosymbiont combinations of the coral model Aiptasia. As expected, all host-endosymbiont combinations showed reduced photosynthetic efficiency and endosymbiont loss during heat stress, but the severity of bleaching was significantly reduced with increasing salinities for one of the host-endosymbiont combinations. Our results show that higher salinities can convey increased thermotolerance in Aiptasia, although this effect seems to be dependent on the particular host strain and/or associated symbiont type. This finding may help explain the extraordinarily high thermotolerance of corals in high salinity environments such as the Red Sea and the Persian/Arabian Gulf and provides novel insight regarding factors that contribute to thermotolerance. Since our results are based on a salinity effect in symbiotic sea anemones, it remains to be determined whether this salinity effect can also be observed in stony corals.

  2. Thermotolerant genes essential for survival at a critical high temperature in thermotolerant ethanologenic Zymomonas mobilis TISTR 548.

    Science.gov (United States)

    Charoensuk, Kannikar; Sakurada, Tomoko; Tokiyama, Amina; Murata, Masayuki; Kosaka, Tomoyuki; Thanonkeo, Pornthap; Yamada, Mamoru

    2017-01-01

    High-temperature fermentation (HTF) technology is expected to reduce the cost of bioconversion of biomass to fuels or chemicals. For stable HTF, the development of a thermotolerant microbe is indispensable. Elucidation of the molecular mechanism of thermotolerance would enable the thermal stability of microbes to be improved. Thermotolerant genes that are essential for survival at a critical high temperature (CHT) were identified via transposon mutagenesis in ethanologenic, thermotolerant Zymomonas mobilis TISTR 548. Surprisingly, no genes for general heat shock proteins except for degP were included. Cells with transposon insertion in these genes showed a defect in growth at around 39 °C but grew normally at 30 °C. Of those, more than 60% were found to be sensitive to ethanol at 30 °C, indicating that the mechanism of thermotolerance partially overlaps with that of ethanol tolerance in the organism. Products of these genes were classified into nine categories of metabolism, membrane stabilization, transporter, DNA repair, tRNA modification, protein quality control, translation control, cell division, and transcriptional regulation. The thermotolerant genes of Escherichia coli and Acetobacter tropicalis that had been identified can be functionally classified into 9 categories according to the classification of those of Z. mobilis, and the ratio of thermotolerant genes to total genomic genes in Z. mobilis is nearly the same as that in E. coli, though the ratio in A. tropicalis is relatively low. There are 7 conserved thermotolerant genes that are shared by these three or two microbes. These findings suggest that Z. mobilis possesses molecular mechanisms for its survival at a CHT that are similar to those in E. coli and A. tropicalis. The mechanisms may mainly contribute to membrane stabilization, protection and repair of damage of macromolecules and maintenance of cellular metabolism at a CHT. Notably, the contribution of heat shock proteins to such survival

  3. High-temperature ethanol production using thermotolerant yeast newly isolated from Greater Mekong Subregion

    Directory of Open Access Journals (Sweden)

    Atiya Techaparin

    Full Text Available Abstract The application of high-potential thermotolerant yeasts is a key factor for successful ethanol production at high temperatures. Two hundred and thirty-four yeast isolates from Greater Mekong Subregion (GMS countries, i.e., Thailand, The Lao People's Democratic Republic (Lao PDR and Vietnam were obtained. Five thermotolerant yeasts, designated Saccharomyces cerevisiae KKU-VN8, KKU-VN20, and KKU-VN27, Pichia kudriavzevii KKU-TH33 and P. kudriavzevii KKU-TH43, demonstrated high temperature and ethanol tolerance levels up to 45 °C and 13% (v/v, respectively. All five strains produced higher ethanol concentrations and exhibited greater productivities and yields than the industrial strain S. cerevisiae TISTR5606 during high-temperature fermentation at 40 °C and 43 °C. S. cerevisiae KKU-VN8 demonstrated the best performance for ethanol production from glucose at 37 °C with an ethanol concentration of 72.69 g/L, a productivity of 1.59 g/L/h and a theoretical ethanol yield of 86.27%. The optimal conditions for ethanol production of S. cerevisiae KKU-VN8 from sweet sorghum juice (SSJ at 40 °C were achieved using the Box-Behnken experimental design (BBD. The maximal ethanol concentration obtained during fermentation was 89.32 g/L, with a productivity of 2.48 g/L/h and a theoretical ethanol yield of 96.32%. Thus, the newly isolated thermotolerant S. cerevisiae KKU-VN8 exhibits a great potential for commercial-scale ethanol production in the future.

  4. High temperature alcoholic fermentation of orange peel by the newly isolated thermotolerant Pichia kudriavzevii KVMP10.

    Science.gov (United States)

    Koutinas, M; Patsalou, M; Stavrinou, S; Vyrides, I

    2016-01-01

    This work explores the potential for the development of orange peel based ethanol bioprocesses through isolation of the thermotolerant Pichia kudriavzevii KVMP10. A model solution of hydrolysed Valencia orange peel was employed to determine the ethanologenic potential of the yeast, which was maximized at 42°C producing 54 g l(-1) of ethanol. The effect of orange peel oil on bioethanol formation was investigated at 30 and 42°C confirming that the minimum inhibitory peel oil content was 0·01% (v/v). Pichia kudriavzevii KVMP10 demonstrated significant technological advantages for the production of sustainable bioenergy, such as utilization of both hexoses (glucose, sucrose, fructose and galactose) and pentoses (xylose) at high temperatures, exemplifying its great potential for application in orange peel based biorefineries for ethanol production. Citrus peel waste is one of the most underutilized and geographically diverse residues in the planet. In attempt to develop a citrus peel based biorefinery we report here the isolation of a yeast which exhibited favourable technological characteristics for the production of ethanol through utilization of the specific food waste. Pichia kudriavzevii KVMP10 was highly thermotolerant and utilized both hexoses and pentoses for ethanol production, which was achieved at elevated rates, highlighting its great potential for application in ethanol production processes from citrus peel. © 2015 The Society for Applied Microbiology.

  5. The Metabolic Basis of Pollen Thermo-Tolerance: Perspectives for Breeding

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    Marine J. Paupière

    2014-09-01

    Full Text Available Crop production is highly sensitive to elevated temperatures. A rise of a few degrees above the optimum growing temperature can lead to a dramatic yield loss. A predicted increase of 1–3 degrees in the twenty first century urges breeders to develop thermo-tolerant crops which are tolerant to high temperatures. Breeding for thermo-tolerance is a challenge due to the low heritability of this trait. A better understanding of heat stress tolerance and the development of reliable methods to phenotype thermo-tolerance are key factors for a successful breeding approach. Plant reproduction is the most temperature-sensitive process in the plant life cycle. More precisely, pollen quality is strongly affected by heat stress conditions. High temperature leads to a decrease of pollen viability which is directly correlated with a loss of fruit production. The reduction in pollen viability is associated with changes in the level and composition of several (groups of metabolites, which play an important role in pollen development, for example by contributing to pollen nutrition or by providing protection to environmental stresses. This review aims to underline the importance of maintaining metabolite homeostasis during pollen development, in order to produce mature and fertile pollen under high temperature. The review will give an overview of the current state of the art on the role of various pollen metabolites in pollen homeostasis and thermo-tolerance. Their possible use as metabolic markers to assist breeding programs for plant thermo-tolerance will be discussed.

  6. Improved ethanol production of a newly isolated thermotolerant Saccharomyces cerevisiae strain after high-energy-pulse-electron beam.

    Science.gov (United States)

    Zhang, Q; Fu, Y; Wang, Y; Han, J; Lv, J; Wang, S

    2012-02-01

    To isolate thermotolerant Saccharomyces cerevisiae with high-energy-pulse-electron (HEPE) beam, to optimize the mutation strain fermentation conditions for ethanol production and to conduct a preliminary investigation into the thermotolerant mechanisms. After HEPE beam radiation, the thermotolerant S. cerevisiae strain Y43 was obtained at 45°C. Moreover, the fermentation conditions of mutant Y43 were optimized by L3(3) orthogonal experiment. The optimal glucose content and initial pH for fermentation were 20% g l(-1) and 4·5, respectively; peptone content was the most neglected important factor. Under this condition, ethanol production of Y43 was 83·1 g l(-1) after fermentation for 48 h at 43°C, and ethanol yield was 0·42 g g(-1), which was about 81·5% of the theoretical yield. The results also showed that the trehalose content and the expression of the genes MSN2, SSA3 and TPS1 in Y43 were higher than those in the original strain (YE0) under the same stress conditions. A genetically stable mutant strain with high ethanol yield under heat stress was obtained using HEPE. This mutant may be a suitable candidate for the industrial-scale ethanol production. High-energy-pulse-electron radiation is a new efficient technology in breeding micro-organisms. The mutant obtained in this work has the advantages in industrial ethanol production under thermostress. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  7. High alcohol production by solid substrate fermentation from starchy substrates using thermotolerant Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Sree, N.K.; Sridhar, M.; Suresh, K.; Rao, L.V. [Department of Microbiology, Osmania University, Hyderabad 500007, Andhra Pradesh (India)

    1999-06-01

    Solid Substrate Fermentation system (SSF) was used to produce ethanol from various starchy substrates like sweet sorghum, sweet potato, wheat flour, rice starch, soluble starch and potato starch using thermotolerant yeast isolate (VS{sub 3}) by simultaneous saccharification and fermentation process. Alcohol produced was estimated by gas chromatography after an incubation time of 96 hrs at 37 C and 42 C. More ethanol was produced from rice starch and sweet sorghum. The maximum amount of ethanol produced from these substrates using VS{sub 3} was 10 g/100 g and 3.5 g/100 g substrate (rice starch) and 8.2 g and 7.5 g/100 g substrate (sweet sorghum) at 37 C and 42 C respectively. (orig.) With 2 figs., 1 tab., 12 refs.

  8. Direct ethanol production from cellulosic materials at high temperature using the thermotolerant yeast Kluyveromyces marxianus displaying cellulolytic enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Yanase, Shuhei; Yamada, Ryosuke; Ogino, Chiaki; Kondo, Akihiko [Kobe Univ. (Japan). Dept. of Chemical Science and Engineering; Hasunuma, Tomohisa; Tanaka, Tsutomu; Fukuda, Hideki [Kobe Univ. (Japan). Organization of Advanced Science and Technology

    2010-09-15

    To exploit cellulosic materials for fuel ethanol production, a microorganism capable of high temperature and simultaneous saccharification-fermentation has been required. However, a major drawback is the optimum temperature for the saccharification and fermentation. Most ethanol-fermenting microbes have an optimum temperature for ethanol fermentation ranging between 28 C and 37 C, while the activity of cellulolytic enzymes is highest at around 50 C and significantly decreases with a decrease in temperature. Therefore, in the present study, a thermotolerant yeast, Kluyveromyces marxianus, which has high growth and fermentation at elevated temperatures, was used as a producer of ethanol from cellulose. The strain was genetically engineered to display Trichoderma reesei endoglucanase and Aspergillus aculeatus {beta}-glucosidase on the cell surface, which successfully converts a cellulosic {beta}-glucan to ethanol directly at 48 C with a yield of 4.24 g/l from 10 g/l within 12 h. The yield (in grams of ethanol produced per gram of {beta}-glucan consumed) was 0.47 g/g, which corresponds to 92.2% of the theoretical yield. This indicates that high-temperature cellulose fermentation to ethanol can be efficiently accomplished using a recombinant K. marxianus strain displaying thermostable cellulolytic enzymes on the cell surface. (orig.)

  9. EVALUATION OF THERMOTOLERANT ACETOBACTER PASTEURIANUS STRAINS ISOLATED FROM MOROCCAN FRUITS CATALYZING OXIDATIVE FERMENTATION AT HIGH TEMPERATURE.

    Science.gov (United States)

    Mounir, M; Shafiei, R; Zarmehrkhorshid, R; Hamouda, A; Alaoui, M Ismaili; Thonart, P

    2015-01-01

    Six strains of acetic acid bacteria were isolated from Moroccan local products and their potential as industrial strains was evaluated in lab-bioreactor. Three of them, namely TAV01, AF01 and CV01, isolated from traditional apple vinegar, apple and cactus fruit, respectively were selected and their responses to high temperature were assessed. Morphological and biochemical identification confirmed that these strains belong to Acetobacter species. Their growth and acetic acid production were compared with the thermoresistant reference strain, Acetobacter senegalensis and mesophilic strains of Acetobacter pasteurianus. The two strains AF01 and CV01 showed abundant growth and noticeable acetic acid production ability at high temperatures (38 to 41°C). A thermophilic character was observed for AF01 strain. Indeed, this bacterium grew better at 38 than 30°C.

  10. Application of thermotolerant microorganisms for biofertilizer preparation.

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    Chen, Kuo-Shu; Lin, Yann-Shying; Yang, Shang-Shyng

    2007-12-01

    Intensive agriculture is practised in Taiwan, and compost application is very popular as a means of improving the soil physical properties and supplying plant nutrition. We tested the potential of inoculation with thermotolerant microorganisms to shorten the maturity and improve the quality of biofertilizer prepared by composting. Thermotolerant microorganisms were isolated from compost and reinoculated for the preparation of biofertilizer. The physical, chemical and biological properties of the biofertilizer were determined during composting. The effects of biofertilizer application on the growth and yield of rape were also studied. Among 3823 colonies of thermotolerant microorganisms, Streptomyces thermonitrificans NTU-88, Streptococcus sp. NTU-130 and Aspergillus fumigatus NTU-132 exhibited high growth rates and cellulolytic and proteolytic activities. When a mixture of rice straw and swine manure were inoculated with these isolates and composted for 61 days, substrate temperature increased initially and then decreased gradually during composting. Substrate pH increased from 7.3 to 8.5. Microbial inoculation enhanced the rate of maturity, and increased the content of ash and total and immobilized nitrogen, improved the germination rate of alfalfa seed, and decreased the content of total organic carbon and the carbon/nitrogen ratio. Biofertilizer application increased the growth and yield of rape. Inoculation of thermotolerant and thermophilic microorganisms to agricultural waste for biofertilizer preparation enhances the rate of maturity and improves the quality of the resulting biofertilizer. Inoculation of appropriate microorganisms in biofertilizer preparation might be usefully applied to agricultural situations.

  11. QTL Analysis of High Thermotolerance with Superior and Downgraded Parental Yeast Strains Reveals New Minor QTLs and Converges on Novel Causative Alleles Involved in RNA Processing

    Science.gov (United States)

    Yang, Yudi; Foulquié-Moreno, Maria R.; Clement, Lieven; Erdei, Éva; Tanghe, An; Schaerlaekens, Kristien; Dumortier, Françoise; Thevelein, Johan M.

    2013-01-01

    Revealing QTLs with a minor effect in complex traits remains difficult. Initial strategies had limited success because of interference by major QTLs and epistasis. New strategies focused on eliminating major QTLs in subsequent mapping experiments. Since genetic analysis of superior segregants from natural diploid strains usually also reveals QTLs linked to the inferior parent, we have extended this strategy for minor QTL identification by eliminating QTLs in both parent strains and repeating the QTL mapping with pooled-segregant whole-genome sequence analysis. We first mapped multiple QTLs responsible for high thermotolerance in a natural yeast strain, MUCL28177, compared to the laboratory strain, BY4742. Using single and bulk reciprocal hemizygosity analysis we identified MKT1 and PRP42 as causative genes in QTLs linked to the superior and inferior parent, respectively. We subsequently downgraded both parents by replacing their superior allele with the inferior allele of the other parent. QTL mapping using pooled-segregant whole-genome sequence analysis with the segregants from the cross of the downgraded parents, revealed several new QTLs. We validated the two most-strongly linked new QTLs by identifying NCS2 and SMD2 as causative genes linked to the superior downgraded parent and we found an allele-specific epistatic interaction between PRP42 and SMD2. Interestingly, the related function of PRP42 and SMD2 suggests an important role for RNA processing in high thermotolerance and underscores the relevance of analyzing minor QTLs. Our results show that identification of minor QTLs involved in complex traits can be successfully accomplished by crossing parent strains that have both been downgraded for a single QTL. This novel approach has the advantage of maintaining all relevant genetic diversity as well as enough phenotypic difference between the parent strains for the trait-of-interest and thus maximizes the chances of successfully identifying additional minor

  12. QTL analysis of high thermotolerance with superior and downgraded parental yeast strains reveals new minor QTLs and converges on novel causative alleles involved in RNA processing.

    Directory of Open Access Journals (Sweden)

    Yudi Yang

    Full Text Available Revealing QTLs with a minor effect in complex traits remains difficult. Initial strategies had limited success because of interference by major QTLs and epistasis. New strategies focused on eliminating major QTLs in subsequent mapping experiments. Since genetic analysis of superior segregants from natural diploid strains usually also reveals QTLs linked to the inferior parent, we have extended this strategy for minor QTL identification by eliminating QTLs in both parent strains and repeating the QTL mapping with pooled-segregant whole-genome sequence analysis. We first mapped multiple QTLs responsible for high thermotolerance in a natural yeast strain, MUCL28177, compared to the laboratory strain, BY4742. Using single and bulk reciprocal hemizygosity analysis we identified MKT1 and PRP42 as causative genes in QTLs linked to the superior and inferior parent, respectively. We subsequently downgraded both parents by replacing their superior allele with the inferior allele of the other parent. QTL mapping using pooled-segregant whole-genome sequence analysis with the segregants from the cross of the downgraded parents, revealed several new QTLs. We validated the two most-strongly linked new QTLs by identifying NCS2 and SMD2 as causative genes linked to the superior downgraded parent and we found an allele-specific epistatic interaction between PRP42 and SMD2. Interestingly, the related function of PRP42 and SMD2 suggests an important role for RNA processing in high thermotolerance and underscores the relevance of analyzing minor QTLs. Our results show that identification of minor QTLs involved in complex traits can be successfully accomplished by crossing parent strains that have both been downgraded for a single QTL. This novel approach has the advantage of maintaining all relevant genetic diversity as well as enough phenotypic difference between the parent strains for the trait-of-interest and thus maximizes the chances of successfully identifying

  13. Enhanced pathway efficiency of Saccharomyces cerevisiae by introducing thermo-tolerant devices.

    Science.gov (United States)

    Liu, Yueqin; Zhang, Genli; Sun, Huan; Sun, Xiangying; Jiang, Nisi; Rasool, Aamir; Lin, Zhanglin; Li, Chun

    2014-10-01

    In this study, thermo-tolerant devices consisting of heat shock genes from thermophiles were designed and introduced into Saccharomyces cerevisiae for improving its thermo-tolerance. Among ten engineered thermo-tolerant yeasts, T.te-TTE2469, T.te-GroS2 and T.te-IbpA displayed over 25% increased cell density and 1.5-4-fold cell viability compared with the control. Physiological characteristics of thermo-tolerant strains revealed that better cell wall integrity, higher trehalose content and enhanced metabolic energy were preserved by thermo-tolerant devices. Engineered thermo-tolerant strain was used to investigate the impact of thermo-tolerant device on pathway efficiency by introducing β-amyrin synthesis pathway, showed 28.1% increased β-amyrin titer, 28-35°C broadened growth temperature range and 72h shortened fermentation period. The results indicated that implanting heat shock proteins from thermophiles to S. cerevisiae would be an efficient approach to improve its thermo-tolerance. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Methanol-inducible promoter of thermotolerant methylotrophic yeast Ogataea thermomethanolica BCC16875 potential for production of heterologous protein at high temperatures.

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    Promdonkoy, Peerada; Tirasophon, Witoon; Roongsawang, Niran; Eurwilaichitr, Lily; Tanapongpipat, Sutipa

    2014-08-01

    Methanol-utilizing metabolism is generally found in methylotrophic yeasts. Several potential promoters regulating enzymes in this pathway have been extensively studied, especially alcohol oxidase. Here, we characterized the alcohol oxidase gene promoter from thermotolerant Ogataea thermomethanolica (OthAOX). This promoter can be induced by methanol, and was shown to regulate expression of phytase up to 45 °C. The pattern of heterologous phytase N-glycosylation depends on the induction temperature. Unlike the AOX promoter from Pichia pastoris, this OthAOX initially turns on the expression of the heterologous protein at the de-repression stage in the presence of glycerol. Full induction of protein is observed when methanol is present. With this methanol-inducible promoter, target protein can be initially produced prior to the induction phase, which would help shorten the time for protein production. Being able to drive protein expression at various temperatures prompts this newly identified AOX promoter to be potential tool for heterologous protein production in high temperature conditions.

  15. Multiple heat priming enhances thermo-tolerance to a later high temperature stress via improving subcellular antioxidant activities in wheat seedlings.

    Science.gov (United States)

    Wang, Xiao; Cai, Jian; Liu, Fulai; Dai, Tingbo; Cao, Weixing; Wollenweber, Bernd; Jiang, Dong

    2014-01-01

    Seedlings of winter wheat (Triticum aestivum L.) were firstly twice heat-primed at 32/24 °C, and subsequently subjected to a more severe high temperature stress at 35/27 °C. The later high temperature stress significantly decreased plant biomass and leaf total soluble sugars concentration. However, plants experienced priming (PH) up-regulated the Rubisco activase B encoding gene RcaB, which was in accordance with the higher photosynthesis rate in relation to the non-primed plants (NH) under the later high temperature stress. In relation to NH, the major chlorophyll a/b-binding protein gene Cab was down-regulated in PH plants, implying a reduction of the light absorption to protect the photosystem II from excitation energy under high temperature stress. At the same time, under the later high temperature stress PH plants showed significantly higher actual photochemical efficiency, indicating an improvement of light use efficiency due to the priming pre-treatment. Under the later high temperature stress, PH could be maintained a better redox homeostasis than NH, as exemplified by the higher activities of superoxide dismutase (SOD) in chloroplasts and glutathione reductase (GR), and of peroxidase (POD) in mitochondria, which contributed to the lower superoxide radical production rate and malondialdehyde concentration in both chloroplasts and mitochondria. The improved antioxidant capacity in chloroplasts and mitochondria was related to the up-regulated expressions of Cu/Zn-SOD, Mn-SOD and GR in PH. Collectively, heat priming effectively improved thermo-tolerance of wheat seedlings subjected to a later high temperature stress, which could be largely ascribed to the enhanced anti-oxidation at the subcellular level. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  16. Microfluidic high viability neural cell separation using viscoelastically tuned hydrodynamic spreading

    DEFF Research Database (Denmark)

    Wu, Zhigang; Hjort, Klas; Wicher, Grzegorz

    2008-01-01

    A high viability microfluidic cell separation technique of high throughput was demonstrated based on size difference continuous mode hydrodynamic spreading with viscoelastic tuning. Using water with fluorescent dye as sample fluid and in parallel introducing as elution a viscoelastic biocompatible...

  17. Microscale Electro-Hydrodynamic Cell Printing with High Viability.

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    He, Jiankang; Zhao, Xiang; Chang, Jinke; Li, Dichen

    2017-12-01

    Cell printing has gained extensive attentions for the controlled fabrication of living cellular constructs in vitro. Various cell printing techniques are now being explored and developed for improved cell viability and printing resolution. Here an electro-hydrodynamic cell printing strategy is developed with microscale resolution (95%). Unlike the existing electro-hydrodynamic cell jetting or printing explorations, insulating substrate is used to replace conventional semiconductive substrate as the collecting surface which significantly reduces the electrical current in the electro-hydrodynamic printing process from milliamperes (>0.5 mA) to microamperes (printed cells. The smallest width of the electro-hydrodynamically printed hydrogel filament is 82.4 ± 14.3 µm by optimizing process parameters. Multiple hydrogels or multilayer cell-laden constructs can be flexibly printed under cell-friendly conditions. The printed cells in multilayer hydrogels kept alive and gradually spread during 7-days culture in vitro. This exploration offers a novel and promising cell printing strategy which might benefit future biomedical innovations such as microscale tissue engineering, organ-on-a-chip systems, and nanomedicine. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Changes in subendocardial viability ratio with acute high-altitude exposure and protective role of acetazolamide.

    Science.gov (United States)

    Salvi, Paolo; Revera, Miriam; Faini, Andrea; Giuliano, Andrea; Gregorini, Francesca; Agostoni, Piergiuseppe; Becerra, Carlos G Ramos; Bilo, Grzegorz; Lombardi, Carolina; O'Rourke, Michael F; Mancia, Giuseppe; Parati, Gianfranco

    2013-04-01

    High-altitude tourism is increasingly frequent, involving also subjects with manifest or subclinical coronary artery disease. Little is known, however, on the effects of altitude exposure on factors affecting coronary perfusion. The aim of our study was to assess myocardial oxygen supply/demand ratio in healthy subjects during acute exposure at high altitude and to evaluate the effect of acetazolamide on this parameter. Forty-four subjects (21 men, age range: 24-59 years) were randomized to double-blind acetazolamide 250 mg bid or placebo. Subendocardial viability ratio and oxygen supply/demand ratio were estimated on carotid artery by means of a validated PulsePen tonometer, at sea level, before and after treatment, and after acute and more prolonged exposure to high altitude (4559 m). On arrival at high altitude, subendocardial viability ratio was reduced in both placebo (from 1.63±0.15 to 1.18±0.17; PSubendocardial viability ratio returned to sea level values (1.65±0.24) after 3 days at high altitude under acetazolamide but remained lower than at sea level under placebo (1.42±0.22; Psubendocardial oxygen supply triggered by exposure to hypobaric hypoxia. Further studies involving also subjects with known or subclinical coronary artery disease are needed to confirm a protective action of acetazolamide on myocardial viability under high-altitude exposure.

  19. Low nutrient availability reduces high-irradiance–induced viability loss inoceanic phytoplankton

    NARCIS (Netherlands)

    Kulk, G.; van de Poll, W.H.; Visser, R.J.W.; Buma, A.G.J.

    2013-01-01

    In situ viability of oceanic phytoplankton may be relatively low in open oceans. This is assumed to be related to the high-irradiance and low-nutrient conditions typical for oligotrophic regions. However, experimental evidence for this phenomenon was not yet available. In the present study, the

  20. Inorganic salts and intracellular polyphosphate inclusions play a role in the thermotolerance of the immunobiotic Lactobacillus rhamnosus CRL 1505.

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    María A Correa Deza

    Full Text Available In this work, the thermotolerance of Lactobacillus rhamnosus CRL1505, an immunobiotic strain, was studied as a way to improve the tolerance of the strain to industrial processes involving heat stress. The strain displayed a high intrinsic thermotolerance (55°C, 20 min; however, after 5 min at 60°C in phosphate buffer a two log units decrease in cell viability was observed. Different heat shock media were tested to improve the cell survival. Best results were obtained in the mediumcontaining inorganic salts (KH2PO4, Na2HPO4, MnSO4, and MgSO4 likely as using 10% skim milk. Flow cytometry analysis evinced 25.0% live cells and a large number of injured cells (59.7% in the inorganic salts medium after heat stress. The morphological changes caused by temperature were visualized by transmission electronic microscopy (TEM. In addition, TEM observations revealed the presence of polyphosphate (polyP granules in the cells under no-stress conditions. A DAPI-based fluorescence technique, adjusted to Gram-positive bacteria for the first time, was used to determine intracellular polyP levels. Results obtained suggest that the high initial polyP content in L. rhamnosus CRL 1505 together with the presence of inorganic salts in the heat shock medium improve the tolerance of the cells to heat shock. To our knowledge, this is the first report giving evidence of the relationship between polyP and inorganic salts in thermotolerance of lactic acid bacteria.

  1. Modeling the impact of high temperatures on microalgal viability and photosynthetic activity.

    Science.gov (United States)

    Béchet, Quentin; Laviale, Martin; Arsapin, Nicolas; Bonnefond, Hubert; Bernard, Olivier

    2017-01-01

    Culture collapse due to high temperatures can significantly impact the profitability of outdoor algal cultivation systems. The objective of this study was to model for the first time the impact of high temperatures on algal activity and viability. Viability measurements on Dunaliella salina cultures were based on cytometry with two fluorescent markers (erythrosine and fluorescein di-acetate), and photosynthetic activity was measured by Pulse Amplitude Modulation (PAM) fluorometry. Kinetic studies revealed that viability and activity losses during exposure to high temperatures could be described by a Weibull model. Both mortality and activity were shown to be functions of the thermal dose received by the algae, defined as the product of duration of exposure to high temperatures and an exponential function of temperature. Simulations at five climatic locations revealed that culture collapse due to high temperatures could impact productivity of D. salina in non-temperature-controlled outdoor photobioreactors by 35 and 40% in arid and Mediterranean climates, respectively. The model developed in this study can be used to forecast the impact of high temperatures on algal biofuel productivity. When coupled with models predicting the temperature of outdoor cultivation systems, this model can also be used to select the best combination of location, system geometry, and algal species to minimize the risks of culture collapse and therefore maximize biofuel productivity.

  2. Thermotolerant fermenting yeasts for simultaneous saccharification fermentation of lignocellulosic biomass

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    Jairam Choudhary

    2016-05-01

    Full Text Available Lignocellulosic biomass is the most abundant renewable source of energy that has been widely explored as second-generation biofuel feedstock. Despite more than four decades of research, the process of ethanol production from lignocellulosic (LC biomass remains economically unfeasible. This is due to the high cost of enzymes, end-product inhibition of enzymes, and the need for cost-intensive inputs associated with a separate hydrolysis and fermentation (SHF process. Thermotolerant yeast strains that can undergo fermentation at temperatures above 40°C are suitable alternatives for developing the simultaneous saccharification and fermentation (SSF process to overcome the limitations of SHF. This review describes the various approaches to screen and develop thermotolerant yeasts via genetic and metabolic engineering. The advantages and limitations of SSF at high temperatures are also discussed. A critical insight into the effect of high temperatures on yeast morphology and physiology is also included. This can improve our understanding of the development of thermotolerant yeast amenable to the SSF process to make LC ethanol production commercially viable.

  3. Identification of Two Thermotolerance-Related Genes in Agaricus bisporus

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    Rong Chen

    2003-01-01

    Full Text Available To characterize thermotolerance-related genes in Agaricus bisporus strain 02, we employed differential display PCR (DD-PCR to analyze total RNA samples extracted from the mycelia grown at different temperatures. Two partial DNA fragments (023-11A and 023-11B were cloned thus far, the expression of which was correlated with the culturing temperature. The sequences of the two DNA fragments were determined and the results showed that the nucleotide sequence of 023-11A was unknown, and 023-11B was highly similar in nucleotide sequence (identities 24 %, positives 45 % to a gene coding for the karyopherin docking complex of the nuclear pore complex of Saccharomyces cerevisiae. It is possible to use the two fragments for further characterization of full-length coding sequences, which can potentially be used for generating new thermotolerant mushroom strains by transgenic technique.

  4. [Influence of high molecular weight polyethylene on viability of osteoblasts and new bone formation].

    Science.gov (United States)

    Ren, Gaohong; Lin, Angru; Pei, Guoxian; Hu, Basheng

    2006-02-01

    To investigate the influence of high molecular weight polyethylene (HMWP) on the viability of osteoblasts and new bone formation in the process of fracture healing, the osteoblasts derived from adult human bone marrow were cultured in HMWP maceration extract and normal culture medium. The viability of the osteoblasts was measured by MTT assay, and the function of the osteoblasts was detected by use of alkaline phosphatase test kit. The locked double-plating (steel plate and HMWP plate) was implanted and fixed at the artificial fracture of distal femur of dogs. Specimens were gained at 3, 6, 9 and 12 weeks postoperatively, examined with macroscopy, microscope and scanning electron microscope (SEM). The results showed that HMWP did no harm to osteoblasts. There is no significant difference in activities of proliferation and alkaline phosphatase between HMWP maceration extract and normal culture medium at each observation time of at 2,4,8, and 14 dyas (P>0. 05). Bone tissue under the implanted HMWP plate manifested no absorption; the new bones formed under the HMWP plate and gradually matured as time went on. It is demonstrated in this study that HMWP has no adverse influence on the viability of osteoblasts and new bone formation and it can be used as internal fixation implant in treating fractures.

  5. Delayed enhancement imaging of myocardial viability: low-dose high-pitch CT versus MRI

    Energy Technology Data Exchange (ETDEWEB)

    Goetti, Robert; Feuchtner, Gudrun; Stolzmann, Paul; Donati, Olivio F.; Frauenfelder, Thomas; Leschka, Sebastian; Alkadhi, Hatem [University Hospital Zurich, Department of Radiology, Zurich (Switzerland); Wieser, Monika; Plass, Andre [University Hospital Zurich, Division of Cardiac and Vascular Surgery, Zurich (Switzerland)

    2011-10-15

    To evaluate the accuracy of high-pitch delayed enhancement (DE) CT for the assessment of myocardial viability with MRI as the reference standard. Twenty-four patients (mean age 66.9 {+-} 9.2 years) with coronary artery disease underwent DE imaging with 128-slice dual-source CT (prospective electrocardiography (ECG)-triggering) and MRI at 1.5 T. Two observers assessed DE transmurality per segment, and measured signal intensity (MRI) or attenuation (CT) in infarcted and healthy myocardium and noise in the left ventricular blood pool for calculating contrast-to-noise ratios (CNR). 75/408 (18.4%) segments in 18/24 patients (75.0%) showed DE in MRI, of which 28 segments in 10/24 (41.7%) patients were non-viable (scar tissue transmurality >50%). Sensitivity, specificity and accuracy of CT for diagnosis of non-viability were 60.7%, 96.8% and 94.4% per segment, and 90.0%, 92.9% and 91.7% per patient. CNR was significantly higher in MR (7.4 {+-} 3.0 vs. 4.6 {+-} 1.5; p = 0.018), and image noise significantly lower (11.6 {+-} 5.7 vs.15.0 {+-} 4.5; p = 0.019). Radiation dose of DECT was 0.89 {+-} 0.07 mSv. CTDE imaging in the high-pitch mode enables myocardial viability assessment at a low radiation dose and good accuracy compared with MR, although associated with a lower CNR and higher noise. (orig.)

  6. Do Multiwell Plate High Throughput Assays Measure Loss of Cell Viability Following Exposure to Genotoxic Agents?

    Directory of Open Access Journals (Sweden)

    Razmik Mirzayans

    2017-08-01

    Full Text Available Cell-based assays in multiwell plates are widely used for radiosensitivity and chemosensitivity assessment with different mammalian cell types. Despite their relative ease of performance, such assays lack specificity as they do not distinguish between the cytostatic (reversible/sustained growth arrest and cytotoxic (loss of viability effects of genotoxic agents. We recently reported studies with solid tumor-derived cell lines demonstrating that radiosensitivity as measured by multiwell plate colorimetric (e.g., XTT and fluorimetric (e.g., CellTiter-Blue assays reflects growth arrest but not loss of viability. Herein we report similar observations with cancer cell lines expressing wild-type p53 (A549 lung carcinoma or mutant p53 (MDA–MB-231 breast carcinoma after treatment with the chemotherapeutic drug cisplatin. Importantly, we show that treatment of cancer cells with concentrations of cisplatin that result in 50% effect (i.e., IC50 in multiwell plate assays trigger the emergence of growth arrested cells that exhibit highly enlarged morphology, remain viable and adherent to the culture dish, and metabolize the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl-tetrazolium bromide (MTT to its formazan derivative. The emergence of markedly enlarged viable cells complicates the interpretation of chemosensitivity data obtained with multiwell plate high throughput assays. Relying solely on IC50 values could be misleading.

  7. The Cytotoxic Role of Intermittent High Glucose on Apoptosis and Cell Viability in Pancreatic Beta Cells

    Directory of Open Access Journals (Sweden)

    Zhen Zhang

    2014-01-01

    Full Text Available Objectives. Glucose fluctuations are both strong predictor of diabetic complications and crucial factor for beta cell damages. Here we investigated the effect of intermittent high glucose (IHG on both cell apoptosis and proliferation activity in INS-1 cells and the potential mechanisms. Methods. Cells were treated with normal glucose (5.5 mmol/L, constant high glucose (CHG (25 mmol/L, and IHG (rotation per 24 h in 11.1 or 25 mmol/L for 7 days. Reactive oxygen species (ROS, xanthine oxidase (XOD level, apoptosis, cell viability, cell cycle, and expression of cyclinD1, p21, p27, and Skp2 were determined. Results. We found that IHG induced more significant apoptosis than CHG and normal glucose; intracellular ROS and XOD levels were more markedly increased in cells exposed to IHG. Cells treated with IHG showed significant decreased cell viability and increased cell proportion in G0/G1 phase. Cell cycle related proteins such as cyclinD1 and Skp2 were decreased significantly, but expressions of p27 and p21 were increased markedly. Conclusions. This study suggested that IHG plays a more toxic effect including both apoptosis-inducing and antiproliferative effects on INS-1 cells. Excessive activation of cellular stress and regulation of cyclins might be potential mechanism of impairment in INS-1 cells induced by IHG.

  8. [Thermophilic and thermotolerant bacteria that assimilate methane].

    Science.gov (United States)

    Malashenko, Iu R; Romanovskaia, V A; Bogachenko, V N; Shved, A D

    1975-01-01

    Microorganisms assimilating methane at temperatures above 40 degrees C were isolated from various natural sources: ooze, mud, waste water of coal pits. The bacteria are obligate methylotrophs and are represented by two groups: (a) thermotolerant, growing at 37 to 45 degrees C; and (b) thermophilic, growing at 50 to 62 degrees C. The selective factor used to isolate various physiological forms of methylotrophs is corresponding temperatures of growth which allow to isolate from the same substrate meso-, thermotolerant, and thermophilic forms. Morphological and physiological properties of the strains are described. The thermotolerant cultures of methylotrophs are similar to Methylobacter vinelandii, though differ from it by some characteristics. The thermophilic microorganisms should be classed as a separate species Methylococcus thermophilus.

  9. Viability of common wine spoilage organisms after exposure to high power ultrasonics.

    Science.gov (United States)

    Luo, Hua; Schmid, Frank; Grbin, Paul R; Jiranek, Vladimir

    2012-05-01

    Microbial spoilage of wine can lead to significant economic loss. At present sulfur dioxide is the main additive to juice/must/wine to prevent and control microbial spoilage. As an alternative, or complement to SO(2), high power ultrasonics (HPU) may be used to control microbes. Several wine spoilage yeasts and bacteria were treated with HPU in saline (0.9% w/v NaCl), juice and red wine to assess their susceptibility to HPU. Significant killing was seen across several yeasts and bacteria commonly associated with winemaking and wine spoilage. In general the viability of yeast was more affected than that of bacteria. Copyright © 2011 Elsevier B.V. All rights reserved.

  10. Enhanced lipid production in thermo-tolerant mutants of Chlorella pyrenoidosa NCIM 2738.

    Science.gov (United States)

    Sachdeva, Neha; Gupta, Ravi Prakash; Mathur, Anshu Shankar; Tuli, Deepak Kumar

    2016-12-01

    The present study aimed to develop thermo-tolerant mutants of Chlorella pyrenoidosa NCIM 2738 for high lipids production. For this, ethyl methane sulfonate was used, which generated two effective thermo-tolerant mutants, M18 and M24 of Chlorella pyrenoidosa NCIM 2738, capable of surviving at temperature up to 47°C and showing improved lipid and biomass yields. They showed 59.62% and 50.75% increase, respectively in lipid content compared to wild type at 30°C, which could not grow at temperature above 35°C. The novelty of this study lied in incorporation of PAM Flurometry with mutagenesis to generate thermo-tolerant mutants of C. pyrenoidosa and investigating the reasons for increased yields of mutants at cellular and photosynthetic levels with the aim to use them for commercial biodiesel production. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Effect of high hydrostatic pressure on mycelial development, spore viability and enzyme activity of Penicillium Roqueforti.

    Science.gov (United States)

    Martínez-Rodríguez, Yamile; Acosta-Muñiz, Carlos; Olivas, Guadalupe I; Guerrero-Beltrán, José; Rodrigo-Aliaga, Dolores; Mujica-Paz, Hugo; Welti-Chanes, Jorge; Sepulveda, David R

    2014-01-03

    This study investigated the effect of high hydrostatic pressure treatments on mycelial development, spore viability, and total proteolytic and lipolytic activity of Penicillium roqueforti PV-LYO 10 D. Fungus growing in liquid medium was pressure-treated at 300, 400, and 500 MPa for 10 min at 20°C following seven days of incubation at 25°C and analyzed periodically up to day 9 after treatments to evaluate the effect on fungal growth. Mycelial mass of P. roqueforti was significantly affected at all pressure treatments evaluated, being 15.48%, 22.28%, 30.03%, and 12.53% lower than controls on day 1, 3, 6, and 9 after 300 MPa treatment, respectively. In a similar way, at 400 and 500 MPa, mycelial mass was 31.08% and 60.34% lower than controls one day after treatments and 49.74% and 80.85% lower on day 9, respectively. The viability of P. roqueforti spores decreased by 36.53% at 300 MPa, and complete inactivation took place at ≥400 MPa from an initial count of 7 log cfu/mL. Total proteolytic activity was not significantly affected at 300 MPa but was reduced by 18.22% at 400 MPa and by 43.18% at 500 MPa. Total lipolytic activity also decreased as the intensity of the pressure treatments increased. 21.69%, 39.12%, and 56.26% activity reductions were observed when treatments of 300, 400 and 500 MPa were applied, respectively. The results from this study show that pressure treatments are able to control growth, inactivate spores, and alter enzyme activity of P. roqueforti, which could be of interest in extending the shelf-life of blue-veined cheeses and other food products. © 2013.

  12. Rewiring of Signaling Networks Modulating Thermotolerance in the Human Pathogen Cryptococcus neoformans.

    Science.gov (United States)

    Yang, Dong-Hoon; Jung, Kwang-Woo; Bang, Soohyun; Lee, Jang-Won; Song, Min-Hee; Floyd-Averette, Anna; Festa, Richard A; Ianiri, Giuseppe; Idnurm, Alexander; Thiele, Dennis J; Heitman, Joseph; Bahn, Yong-Sun

    2017-01-01

    Thermotolerance is a crucial virulence attribute for human pathogens, including the fungus Cryptococcus neoformans that causes fatal meningitis in humans. Loss of the protein kinase Sch9 increases C. neoformans thermotolerance, but its regulatory mechanism has remained unknown. Here, we studied the Sch9-dependent and Sch9-independent signaling networks modulating C. neoformans thermotolerance by using genome-wide transcriptome analysis and reverse genetic approaches. During temperature upshift, genes encoding for molecular chaperones and heat shock proteins were upregulated, whereas those for translation, transcription, and sterol biosynthesis were highly suppressed. In this process, Sch9 regulated basal expression levels or induced/repressed expression levels of some temperature-responsive genes, including heat shock transcription factor (HSF1) and heat shock proteins (HSP104 and SSA1). Notably, we found that the HSF1 transcript abundance decreased but the Hsf1 protein became transiently phosphorylated during temperature upshift. Nevertheless, Hsf1 is essential for growth and its overexpression promoted C. neoformans thermotolerance. Transcriptome analysis using an HSF1 overexpressing strain revealed a dual role of Hsf1 in the oxidative stress response and thermotolerance. Chromatin immunoprecipitation demonstrated that Hsf1 binds to the step-type like heat shock element (HSE) of its target genes more efficiently than to the perfect- or gap-type HSE. This study provides insight into the thermotolerance of C. neoformans by elucidating the regulatory mechanisms of Sch9 and Hsf1 through the genome-scale identification of temperature-dependent genes. Copyright © 2017 by the Genetics Society of America.

  13. Cell viability and MRI performance of highly efficient polyol-coated magnetic nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Arteaga-Cardona, Fernando [Universidad de las Américas de Puebla, Departamento de Ciencias Químico-Biológicas (Mexico); Gutiérrez-García, Eric [Instituto Literario, Universidad Autónoma del Estado de México (Mexico); Hidalgo-Tobón, Silvia, E-mail: shid@xanum.uam.mx [Universidad Autónoma Metropolitana, Departamento de Física (Mexico); López-Vasquez, Ciro; Brito-Barrera, Yazmín A. [Universidad de las Américas de Puebla, Departamento de Ciencias Químico-Biológicas (Mexico); Flores-Tochihuitl, Julia [Benemérita Universidad Autónoma de Puebla, Facultad de Estomatología (Mexico); Angulo-Molina, Aracely [Universidad de Sonora, Departamento de Ciencias Químico-Biológicas (Mexico); Reyes-Leyva, Julio R. [Instituto Mexicano del Seguro Social, Centro de Investigación Biomédica de Oriente (CIBIOR) (Mexico); González-Rodríguez, Roberto; Coffer, Jeffery L. [Texas Christian University, Department of Chemistry (United States); Pal, Umapada [Benemérita Universidad Autónoma de Puebla, Apdo, Instituto de Física (Mexico); and others

    2016-11-15

    This work aimed at determining conditions that would allow us to control the size of the NPs and create a system with characteristics apt for biomedical applications. We describe a comprehensive study on the synthesis and physical characterization of two highly sensitive sets of triethylene glycol (TREG) and polyethylene glycol (PEG)-coated superparamagnetic iron oxide nanoparticles (SPIONs) to be evaluated for use as magnetic resonance (MR) contrast agents. The ferrofluids demonstrated excellent colloidal stability in deionized water at pH 7.0 as indicated by dynamic light scattering (DLS) data. The magnetic relaxivities, r{sub 2}, were measured on a 1.5 T clinical MRI instrument. Values in the range from 205 to 257 mM{sup −1} s{sup −1} were obtained, varying proportionally to the SPIONs’ sizes and coating nature. Further in vitro cell viability tests and in vivo biodistribution analyses of the intravenously administered nanoparticles showed that the prepared systems have good biocompatibility and migrate to several organs, mainly the meninges, spleen, and liver. Based on these results, our findings demonstrated the potential utility of these nanosystems as clinical contrast agents for MR imaging.

  14. Viability Theory

    CERN Document Server

    Aubin, Jean-Pierre; Saint-Pierre, Patrick

    2011-01-01

    Viability theory designs and develops mathematical and algorithmic methods for investigating the adaptation to viability constraints of evolutions governed by complex systems under uncertainty that are found in many domains involving living beings, from biological evolution to economics, from environmental sciences to financial markets, from control theory and robotics to cognitive sciences. It involves interdisciplinary investigations spanning fields that have traditionally developed in isolation. The purpose of this book is to present an initiation to applications of viability theory, explai

  15. High-content assays for characterizing the viability and morphology of 3D cancer spheroid cultures.

    Science.gov (United States)

    Sirenko, Oksana; Mitlo, Trisha; Hesley, Jayne; Luke, Steve; Owens, Windsor; Cromwell, Evan F

    2015-09-01

    There is an increasing interest in using three-dimensional (3D) spheroids for modeling cancer and tissue biology to accelerate translation research. Development of higher throughput assays to quantify phenotypic changes in spheroids is an active area of investigation. The goal of this study was to develop higher throughput high-content imaging and analysis methods to characterize phenotypic changes in human cancer spheroids in response to compound treatment. We optimized spheroid cell culture protocols using low adhesion U-bottom 96- and 384-well plates for three common cancer cell lines and improved the workflow with a one-step staining procedure that reduces assay time and minimizes variability. We streamlined imaging acquisition by using a maximum projection algorithm that combines cellular information from multiple slices through a 3D object into a single image, enabling efficient comparison of different spheroid phenotypes. A custom image analysis method was implemented to provide multiparametric characterization of single-cell and spheroid phenotypes. We report a number of readouts, including quantification of marker-specific cell numbers, measurement of cell viability and apoptosis, and characterization of spheroid size and shape. Assay performance was assessed using established anticancer cytostatic and cytotoxic drugs. We demonstrated concentration-response effects for different readouts and measured IC50 values, comparing 3D spheroid results to two-dimensional cell cultures. Finally, a library of 119 approved anticancer drugs was screened across a wide range of concentrations using HCT116 colon cancer spheroids. The proposed methods can increase performance and throughput of high-content assays for compound screening and evaluation of anticancer drugs with 3D cell models.

  16. Pleurotus sajor-caju HSP100 complements a thermotolerance defect in hsp104 mutant Saccharomyces cerevisiae.

    Science.gov (United States)

    Lee, Jin-Ohk; Jeong, Mi-Jeong; Kwon, Tack-Ryun; Lee, Seung-Kon; Byun, Myung-Ok; Chung, Ill-Min; Park, Soo-Chul

    2006-06-01

    A putative Hsp100 gene was cloned from the fungus Pleurotus sajor-caju. mRNA expression studies demonstrated that this gene (designated PsHsp100) is highly induced by high temperature,induced less strongly by exposure to ethanol, and not induced by drought or salinity. Heat shock induction is detectable at 37 degrees C and reaches a maximum level at 42 degrees C. PsHsp100 mRNA levels sharply increased within 15 min of exposure to high temperature, and reached a maximum expression level at 2 h that was maintained for several hours. These results indicate that PsHsp100 could work at an early step in thermotolerance. To examine its function, PsHsp100 was transformed into a temperature-sensitive hsp104 deletion mutant Saccharomycetes cerivisiae strain to test the hypothesis that PsHSP100 is an protein that functions in thermotolerance. Overexpression of PsHSP100 complemented the thermotolerance defect of the hsp104 mutant yeast, allowing them being survive even at 50 degree C for 4 h. These results indicate that PsHSP100 protein is functional as an HSP100 in yeast and could play and important role in thermotolerance in P. sajor-caju.

  17. Isolation, cloning and molecular characterization of a thermotolerant ...

    African Journals Online (AJOL)

    Isolation, cloning and molecular characterization of a thermotolerant xylanase from Streptomyces sp. THW31. Thayat Sriyapai, Peechapack Somyoonsap, Supatra Areekit, Paisarn Khawsak, Arda Pakpitcharoen, Kosum Chansiri ...

  18. Divergent distribution of the sensor kinase CosS in non-thermotolerant campylobacter species and its functional incompatibility with the response regulator CosR of Campylobacter jejuni.

    Science.gov (United States)

    Hwang, Sunyoung; Miller, William G; Ryu, Sangryeol; Jeon, Byeonghwa

    2014-01-01

    Two-component signal transduction systems are commonly composed of a sensor histidine kinase and a cognate response regulator, modulating gene expression in response to environmental changes through a phosphorylation-dependent process. CosR is an OmpR-type response regulator essential for the viability of Campylobacter jejuni, a major foodborne pathogenic species causing human gastroenteritis. Although CosR is a response regulator, its cognate sensor kinase has not been identified in C. jejuni. In this study, DNA sequence analysis of the cosR flanking regions revealed that a gene encoding a putative sensor kinase, which we named cosS, is prevalent in non-thermotolerant Campylobacter spp., but not in thermotolerant campylobacters. Phosphorylation assays indicated that C. fetus CosS rapidly autophosphorylates and then phosphorylates C. fetus CosR, suggesting that the CosRS system constitutes a paired two-component signal transduction system in C. fetus. However, C. fetus CosS does not phosphorylate C. jejuni CosR, suggesting that CosR may have different regulatory cascades between thermotolerant and non-thermotolerant Campylobacter species. Comparison of CosR homolog amino acid sequences showed that the conserved phosphorylation residue (D51), which is present in all non-thermotolerant Campylobacter spp., is absent from the CosR homologs of thermotolerant Campylobacter species. However, C. jejuni CosR was not phosphorylated by C. fetus CosS even after site-directed mutagenesis of N51D, implying that C. jejuni CosR may possibly function phosphorylation-independently. In addition, the results of cosS mutational analysis indicated that CosS is not associated with the temperature dependence of the Campylobacter spp. despite its unique divergent distribution only in non-thermotolerant campylobacters. The findings in this study strongly suggest that thermotolerant and non-thermotolerant Campylobacter spp. have different signal sensing mechanisms associated with the Cos

  19. Multiple heat priming enhances thermo-tolerance to a later high temperature stress via improving subcellular antioxidant activities in wheat seedlings

    DEFF Research Database (Denmark)

    Wang, Xiao; Cai, Jian; Liu, Fulai

    2014-01-01

    Seedlings of winter wheat (Triticum aestivum L.) were firstly twice heat-primed at 32/24 °C, and subsequently subjected to a more severe high temperature stress at 35/27 °C. The later high temperature stress significantly decreased plant biomass and leaf total soluble sugars concentration. However......, plants experienced priming (PH) up-regulated the Rubisco activase B encoding gene RcaB, which was in accordance with the higher photosynthesis rate in relation to the non-primed plants (NH) under the later high temperature stress. In relation to NH, the major chlorophyll a/b-binding protein gene Cab...... an improvement of light use efficiency due to the priming pre-treatment. Under the later high temperature stress, PH could be maintained a better redox homeostasis than NH, as exemplified by the higher activities of superoxide dismutase (SOD) in chloroplasts and glutathione reductase (GR), and of peroxidase (POD...

  20. 3D Printed Vascular Networks Enhance Viability in High-Volume Perfusion Bioreactor.

    Science.gov (United States)

    Ball, Owen; Nguyen, Bao-Ngoc B; Placone, Jesse K; Fisher, John P

    2016-12-01

    There is a significant clinical need for engineered bone graft substitutes that can quickly, effectively, and safely repair large segmental bone defects. One emerging field of interest involves the growth of engineered bone tissue in vitro within bioreactors, the most promising of which are perfusion bioreactors. Using bioreactor systems, tissue engineered bone constructs can be fabricated in vitro. However, these engineered constructs lack inherent vasculature and once implanted, quickly develop a necrotic core, where no nutrient exchange occurs. Here, we utilized COMSOL modeling to predict oxygen diffusion gradients throughout aggregated alginate constructs, which allowed for the computer-aided design of printable vascular networks, compatible with any large tissue engineered construct cultured in a perfusion bioreactor. We investigated the effect of 3D printed macroscale vascular networks with various porosities on the viability of human mesenchymal stem cells in vitro, using both gas-permeable, and non-gas permeable bioreactor growth chamber walls. Through the use of 3D printed vascular structures in conjunction with a tubular perfusion system bioreactor, cell viability was found to increase by as much as 50% in the core of these constructs, with in silico modeling predicting construct viability at steady state.

  1. Thermotolerance of human myometrium: implications for minimally invasive uterine therapies

    Science.gov (United States)

    Thomas, Aaron C.; Grisez, Brian T.; McMillan, Kathleen; Chill, Nicholas; Harclerode, Tyler P.; Radabaugh, Rebecca; Jones, Ryan M.; Coad, James E.

    2013-02-01

    Endometrial ablation has gained significant clinical acceptance over the last decade as a minimally invasive treatment for abnormal uterine bleeding. To improve upon current thermal injury modeling, it is important to better characterize the myometrium's thermotolerance. The extent of myometrial thermal injury was determined across a spectrum of thermal histories/doses (time-temperature combinations). Fresh extirpated human myometrium was obtained from 13 subjects who underwent a previous scheduled benign hysterectomy. Within two hours of hysterectomy, the unfixed myometrium was treated in a stabilized saline bath with temperatures ranging from 45-70 °C and time intervals from 30- 150 seconds. The time-temperature combinations were selected to simulate treatment times under 2.5 minutes. A total of six such thermal matrices, each comprised of 45 time-temperature combinations, were prepared for evaluation. The treated myometrium was cryosectioned for nitro blue tetrazolium (NBT) staining to assess for thermal respiratory enzyme inactivation. Image analysis was subsequently used to quantitatively assess the stained myometrium's capacity to metabolize the tetrazolium at each time-temperature combination. This colorimetric data was then used as marker of cellular viability and determine survival parameters with implications for developing minimally invasive uterine therapies.

  2. Pleurotus sajor-caju HSP100 complements a thermotolerance ...

    Indian Academy of Sciences (India)

    To examine its function, PsHsp100 was transformed into a temperature-sensitive hsp104 deletion mutant Saccharomyces cerevisiae strain to test the hypothesis that PsHSP100 is an protein that functions in thermotolerance. Overexpression of PsHSP100 complemented the thermotolerance defect of the hsp104 mutant ...

  3. Para-aminobenzoic acid (PABA synthase enhances thermotolerance of mushroom Agaricus bisporus.

    Directory of Open Access Journals (Sweden)

    Zhonglei Lu

    Full Text Available Most mushrooms are thermo-sensitive to temperatures over 23°C, which greatly restricts their agricultural cultivation. Understanding mushroom's innate heat-tolerance mechanisms may facilitate genetic improvements of their thermotolerance. Agaricus bisporus strain 02 is a relatively thermotolerant mushroom strain, while strain 8213 is quite thermo-sensitive. Here, we compared their responses at proteomic level to heat treatment at 33°C. We identified 73 proteins that are differentially expressed between 02 and 8213 or induced upon heat stress in strain 02 itself, 48 of which with a known identity. Among them, 4 proteins are constitutively more highly expressed in 02 than 8213; and they can be further upregulated in response to heat stress in 02, but not in 8213. One protein is encoded by the para-aminobenzoic acid (PABA synthase gene Pabs, which has been shown to scavenge the reactive oxygen species in vitro. Pabs mRNA and its chemical product PABA show similar heat stress induction pattern as PABA synthase protein and are more abundant in 02, indicating transcriptional level upregulation of Pabs upon heat stress. A specific inhibitor of PABA synthesis impaired thermotolerance of 02, while exogenous PABA or transgenic overexpression of 02 derived PABA synthase enhanced thermotolerance of 8213. Furthermore, compared to 8213, 02 accumulated less H2O2 but more defense-related proteins (e.g., HSPs and Chitinase under heat stress. Together, these results demonstrate a role of PABA in enhancing mushroom thermotolerance by removing H2O2 and elevating defense-related proteins.

  4. Perspectives on deciphering mechanisms underlying plant heat stress response and thermotolerance

    Directory of Open Access Journals (Sweden)

    Kamila Lucia Bokszczanin

    2013-08-01

    Full Text Available Global warming is a major threat for agriculture and food safety and in many cases the negative effects are already apparent. The current challenge of basic and applied plant science is to decipher the molecular mechanisms of heat stress response and thermotolerance in detail and use this information to identify genotypes that will withstand unfavorable environmental conditions. Nowadays X-omics approaches complement the findings of previous targeted studies and highlight the complexity of heat stress response mechanisms giving information for so far unrecognized genes, proteins and metabolites as potential key players of thermotolerance. Even more, roles of epigenetic mechanisms and the involvement of small RNAs in thermotolerance are currently emerging and thus open new directions of yet unexplored areas of plant heat stress response. In parallel it is emerging that although the whole plant is vulnerable to heat, specific organs are particularly sensitive to elevated temperatures. This has redirected research from the vegetative to generative tissues. The sexual reproduction phase is considered as the most sensitive to heat and specifically pollen exhibits the highest sensitivity and frequently an elevation of the temperature just a few degrees above the optimum during pollen development can have detrimental effects for crop production. Compared to our knowledge on heat stress response of vegetative tissues, the information on pollen is still scarce. Nowadays, several techniques for high-throughput X-omics approaches provide major tools to explore the principles of pollen heat stress response and thermotolerance mechanisms in specific genotypes. The collection of such information will provide an excellent support for improvement of breeding programs to facilitate the development of tolerant cultivars. The review aims at describing the current knowledge of thermotolerance mechanisms and the technical advances which will foster new insights into

  5. L-Citrulline acts as potential hypothermic agent to afford thermotolerance in chicks.

    Science.gov (United States)

    Chowdhury, Vishwajit S; Han, Guofeng; Bahry, Mohammad A; Tran, Phuong V; Do, Phong H; Yang, Hui; Furuse, Mitsuhiro

    2017-10-01

    Recently we demonstrated that L-citrulline (L-Cit) causes hypothermia in chicks. However, the question of how L-Cit mediates hypothermia remained elusive. Thus, the objective of this study was to examine some possible factors in the process of L-Cit-mediated hypothermia and to confirm whether L-Cit can also afford thermotolerance in young chicks. Chicks were subjected to oral administration of L-Cit along with intraperitoneal injection of a nitric oxide synthase (NOS) inhibitor, NG-nitro-l-arginine methyl ester HCl (L-NAME), to examine the involvement of NO in the process of hypothermia. Food intake and plasma metabolites were also analyzed after oral administration of L-Cit in chicks. To examine thermotolerance, chicks were orally administered with a single dose of L-Cit (15mmol/10ml/kg body weight) or the same dose twice within a short interval of 1h (dual oral administration) before the exposure to high ambient temperature (35 ± 1°C) for 180min. Although the rectal temperature was reduced following administration of L-Cit, L-NAME caused a greater reduction. L-NAME reduced total NO2 and NO3 (NOx) in plasma, which confirmed its inhibitory effect on NO. A single oral administration of L-Cit mediated a persistent state of hypothermia for the 300min of the study without affecting food intake. It was further found that plasma glucose was significantly lower in L-Cit-treated chicks. Dual oral administration of L-Cit, but not a single oral administration, afforded thermotolerance without a significant change in plasma NOx in chicks. In conclusion, our results suggest that L-Cit-mediated hypothermia and thermotolerance may not be involved in NO production. L-Cit-mediated thermotolerance further suggests that L-Cit may serve as an important nutritional supplement that could help in coping with summer heat. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Hsp27 protects mitochondria of thermotolerant cells against apoptotic stimuli

    Science.gov (United States)

    Samali, Afshin; Robertson, John D.; Peterson, Elisabeth; Manero, Florence; van Zeijl, Leone; Paul, Catherine; Cotgreave, Ian A.; Arrigo, André-Patrick; Orrenius, Sten

    2001-01-01

    Enhanced cell survival and resistance to apoptosis during thermotolerance correlates with an increased expression of heat shock proteins (Hsps). Here we present additional evidence in support of the hypothesis that the induction of Hsp27 and Hsp72 during acquired thermotolerance in Jurkat T-lymphocytes prevents apoptosis. In thermotolerant cells, Hsp27 was shown to associate with the mitochondrial fraction, and inhibition of Hsp27 induction during thermotolerance in cells transfected with hsp27 antisense potentiated mitochondrial cytochrome c release after exposure to various apoptotic stimuli, despite the presence of elevated levels of Hsp72. Caspase activation and apoptosis were inhibited under these conditions. In vitro studies revealed that recombinant Hsp72 more efficiently blocked cytochrome c–mediated caspase activation than did recombinant Hsp27. A model is presented for the inhibition of apoptosis during thermotolerance in which Hsp27 preferentially blocks mitochondrial cytochrome c release, whereas Hsp72 interferes with apoptosomal caspase activation. PMID:11525243

  7. High Cell Density Process for Constitutive Production of a Recombinant Phytase in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica Using Table Sugar as Carbon Source.

    Science.gov (United States)

    Charoenrat, Theppanya; Antimanon, Sompot; Kocharin, Kanokarn; Tanapongpipat, Sutipa; Roongsawang, Niran

    2016-12-01

    The yeast Ogataea thermomethanolica has recently emerged as a potential host for heterologous protein expression at elevated temperature. To evaluate the feasibility of O. thermomethanolica as heterologous host in large-scale fermentation, constitutive production of fungal phytase was investigated in fed-batch fermentation. The effect of different temperatures, substrate feeding strategies, and carbon sources on phytase production was investigated. It was found that O. thermomethanolica can grow in the temperature up to 40 °C and optimal at 34 °C. However, the maximum phytase production was observed at 30 °C and slightly decreased at 34 °C. The DOT stat control was the most efficient feeding strategy to obtain high cell density and avoid by-product formation. The table sugar can be used as an alternative substrate for phytase production in O. thermomethanolica. The highest phytase activity (134 U/mL) was obtained from table sugar at 34 °C which was 20-fold higher than batch culture (5.7 U/mL). At a higher cultivation temperature of 38 °C, table sugar can be used as a low-cost substrate for the production of phytase which was expressed with an acceptable yield (85 U/mL). Lastly, the results from this study reveal the industrial favorable benefits of employing O. thermomethanolica as a host for heterologous protein production.

  8. Selection and characterization of a newly isolated thermotolerant Pichia kudriavzevii strain for ethanol production at high temperature from cassava starch hydrolysate.

    Science.gov (United States)

    Yuangsaard, Napatchanok; Yongmanitchai, Wichien; Yamada, Mumoru; Limtong, Savitree

    2013-03-01

    Pichia kudriavzevii DMKU 3-ET15 was isolated from traditional fermented pork sausage by an enrichment technique in a yeast extract peptone dextrose (YPD) broth, supplemented with 4 % (v/v) ethanol at 40 °C and selected based on its ethanol fermentation ability at 40 °C in YPD broth composed of 16 % glucose, and in a cassava starch hydrolysate medium composed of cassava starch hydrolysate adjusted to 16 % glucose. The strain produced ethanol from cassava starch hydrolysate at a high temperature up to 45 °C, but the optimal temperature for ethanol production was at 40 °C. Ethanol production by this strain using shaking flask cultivation was the highest in a medium containing cassava starch hydrolysate adjusted to 18 % glucose, 0.05 % (NH(4))(2)SO(4), 0.09 % yeast extract, 0.05 % KH(2)PO(4), and 0.05 % MgSO(4)·7H(2)O, with a pH of 5.0 at 40 °C. The highest ethanol concentration reached 7.86 % (w/v) after 24 h, with productivity of 3.28 g/l/h and yield of 85.4 % of the theoretical yield. At 42 °C, ethanol production by this strain became slightly lower, while at 45 °C only 3.82 % (w/v) of ethanol, 1.27 g/l/h productivity and 41.5 % of the theoretical yield were attained. In a study on ethanol production in a 2.5-l jar fermenter with an agitation speed of 300 rpm and an aeration rate of 0.1 vvm throughout the fermentation, P. kudriavzevii DMKU 3-ET15 yielded a final ethanol concentration of 7.35 % (w/v) after 33 h, a productivity of 2.23 g/l/h and a yield of 79.9 % of the theoretical yield.

  9. Advanced high-power pulsed light device to decontaminate food from pathogens: effects on Salmonella typhimurium viability in vitro.

    Science.gov (United States)

    Luksiene, Z; Gudelis, V; Buchovec, I; Raudeliuniene, J

    2007-11-01

    The aim of this study was to construct an advanced high-power pulsed light device for decontamination of food matrix and to evaluate its antibacterial efficiency. Key parameters of constructed device-emitted light spectrum, pulse duration, pulse power density, frequency of pulses, dependence of emitted spectrum on input voltage, irradiation homogenicity, possible thermal effects as well as antimicrobial efficiency were evaluated. Antimicrobial efficiency of high-power pulsed light technique was demonstrated and evaluated by two independent methods - spread plate and Miles-Misra method. Viability of Salmonella typhimurium as function of a given light dose (number of pulses) and pulse frequency was examined. According to the data obtained, viability of Salmonella typhimurium reduced by 7 log order after 100 light pulses with power density 133 W cm(-2). In addition, data indicate, that the pulse frequency did not influence the outcome of pathogen inactivation in the region 1-5 Hz. Moreover, no hyperthermic effect was detected during irradiation even after 500 pulses on all shelves with different distance from light source and subsequently different pulse power density (0-252 W cm(-2)). Newly constructed high-power pulsed light technique is effective nonthermal tool for inactivation of Salmonella typhimurium even by 7 log order in vitro. Novel advanced high-power pulsed light device can be a useful tool for development of nonthermal food decontamination technologies.

  10. Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability.

    Directory of Open Access Journals (Sweden)

    Emily Peak

    2010-07-01

    Full Text Available Schistosomiasis, caused by infection with the blood fluke Schistosoma, is responsible for greater than 200,000 human deaths per annum. Objective high-throughput screens for detecting novel anti-schistosomal targets will drive 'genome to drug' lead translational science at an unprecedented rate. Current methods for detecting schistosome viability rely on qualitative microscopic criteria, which require an understanding of parasite morphology, and most importantly, must be subjectively interpreted. These limitations, in the current state of the art, have significantly impeded progress into whole schistosome screening for next generation chemotherapies.We present here a microtiter plate-based method for reproducibly detecting schistosomula viability that takes advantage of the differential uptake of fluorophores (propidium iodide and fluorescein diacetate by living organisms. We validate this high-throughput system in detecting schistosomula viability using auranofin (a known inhibitor of thioredoxin glutathione reductase, praziquantel and a range of small compounds with previously-described (gambogic acid, sodium salinomycin, ethinyl estradiol, fluoxetidine hydrochloride, miconazole nitrate, chlorpromazine hydrochloride, amphotericin b, niclosamide or suggested (bepridil, ciclopirox, rescinnamine, flucytosine, vinblastine and carbidopa anti-schistosomal activities. This developed method is sensitive (200 schistosomula/well can be assayed, relevant to industrial (384-well microtiter plate compatibility and academic (96-well microtiter plate compatibility settings, translatable to functional genomics screens and drug assays, does not require a priori knowledge of schistosome biology and is quantitative.The wide-scale application of this fluorescence-based bioassay will greatly accelerate the objective identification of novel therapeutic lead targets/compounds to combat schistosomiasis. Adapting this bioassay for use with other parasitic worm species

  11. Mechanism of action of hydrogen peroxide in wheat thermotolerance

    African Journals Online (AJOL)

    Mechanism of action of hydrogen peroxide in wheat thermotolerance - interaction between antioxidant isoenzymes, proline and cell membrane. Ranjeet Ranjan Kumar, Sushil Kumar Sharma, Kritika A. Gadpayle, Khushboo Singh, R Sivaranjani, Suneha Goswami, D. Rai Raj ...

  12. Isolation and characterization of thermotolerant ethanol-fermenting ...

    African Journals Online (AJOL)

    Isolation and characterization of thermotolerant ethanol-fermenting yeasts from Laos and application of whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) analysis for their quick identification.

  13. [On the role of gene of SER-4 serotonin receptor in thermotolerance of Caenorhabditis elegans behavior].

    Science.gov (United States)

    Kalinnikova, T B; Kolsanova, R R; Shagidullin, R R; Osipova, E B; Gaĭnutdinov, M Kh

    2013-03-01

    Serotonin reduces the behavior tolerance of Caenorhabditis elegans of the N2 wild-type strain (swimming induced by the mechanical stimulus) to a temperature of 36 degrees C. The sensitivity to the serotonin influence on the behavior thermotolerance remains intact in strains with null mutations of mod-1 (ok103) and ser-1 (ok345) serotonin receptor genes, and is almost completely lost in the ser-4 (ok512) strain with null mutation in the gene of the SER-4 serotonin receptor, which is a homologue of 5-HT1 mammalian serotonin receptor. In addition, nematodes of the ser-4 (ok512) strain have high behavior thermotolerance in the absence of the exogenous serotonin compared to the N2 strain. These data indicate the involvement of the ser-4 gene in the serotonin regulation of the tolerance of C. elegance nervous system functions to hyperthermia.

  14. Pleurotus sajor-caju HSP100 complements a thermotolerance ...

    Indian Academy of Sciences (India)

    Madhu

    functional as an HSP100 in yeast and could play an important role in thermotolerance in P. sajor-caju. [Lee J-O, Jeong M-J, Kwon T-R, Lee S-K, Byun M-O, Chung I-M and Park S-C 2006 Pleurotus sajor-caju HSP100 complements a thermotolerance defect in hsp104 mutant Saccharomyces cerevisiae; J. Biosci. 31 223–233].

  15. Characterization and gene expression profiles of thermotolerant Saccharomyces cerevisiae isolates from Thai fruits.

    Science.gov (United States)

    Auesukaree, Choowong; Koedrith, Preeyaporn; Saenpayavai, Pornpon; Asvarak, Thipa; Benjaphokee, Suthee; Sugiyama, Minetaka; Kaneko, Yoshinobu; Harashima, Satoshi; Boonchird, Chuenchit

    2012-08-01

    For industrial applications, fermentation of ethanol at high temperature offers advantages such as reduction in cooling costs, reduced risk of microbial contamination and higher efficiency of fermentation processes including saccharification and continuous ethanol stripping. Three thermotolerant Saccharomyces cerevisiae isolates (C3723, C3751 and C3867) from Thai fruits were capable of growing and producing 38 g/L ethanol up to 41°C. Based on genetic analyses, these isolates were prototrophic and homothallic, with dominant homothallic and thermotolerant phenotypes. After short-term (30 min) and long-term (12 h) exposure at 37°C, expression levels increased for the heat stress-response genes HSP26, SSA4, HSP82, and HSP104 encoding the heat shock proteins small HSP, HSP70, HSP90 and the HSP100 family, respectively. In isolates C3723 and C3867, expression was significantly higher than that in reference isolates W303 and TISTR5606 for TPS1 encoding trehalose-6-phosphate synthase, NTH1 encoding neutral trehalase and GSY1 encoding glycogen synthase. The results suggested that continuous high expression of heat stress-response genes was important for the long-term, heat stress tolerance of these thermotolerant isolates. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  16. L-arginine, an active component of salmon milt nucleoprotein, promotes thermotolerance via Sirtuin in Caenorhabditis elegans.

    Science.gov (United States)

    Furuhashi, Tsubasa; Matsunaga, Masaji; Asahara, Yuji; Sakamoto, Kazuichi

    2016-03-25

    We previously showed that salmon milt nucleoprotein (NP) promotes thermotolerance in Caenorhabditis elegans; however, the active component and physiological mechanism of this effect has remained unclear. l-arginine (AR) is a major component of protamine and thus it has been proposed as the possible active component of NP. In this study, the viability of C. elegans treated with AR under heat stress was assessed and AR was shown to extend the survival term of the heat-stressed organisms. Additionally, AR was shown to restore the thrashing movement of the worms that is suppressed by heat stress. Treatment with AR was furthermore shown to promote thermotolerance in a DAF-16- and SIR-2.1-dependent manner, where DAF-16 and SIR-2.1 are homologs of FoxO and SirT1, respectively. Taken together, these data suggest that AR is one of the active components of NP and promotes thermotolerance via the activation of DAF-16 and SIR-2.1. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Antiretrovirals for low income countries: an analysis of the commercial viability of a highly competitive market.

    Science.gov (United States)

    Nakakeeto, Olive N; Elliott, Brian V

    2013-02-15

    The price of antiretroviral drugs (ARVs) in low income countries declined steadily in recent years. This raises concerns about the commercial viability of the market of ARVs in low income countries. Using 2 costing scenarios, we modeled the production cost of the most commonly used ARVs in low income countries in 2010 and 2012, and assessed whether, at the median price paid by low income countries, their manufacturers would still make profits. By interviews we consulted 11 generic manufacturers on the current state of the ARV market, and on what would be required to ensure their continued commitment to supply ARVs to low income countries. Using the lowest prices for active pharmaceutical ingredients (API) quoted to WHO, and applying published assumptions about the production cost of ARVs, our baseline estimate was that Indian generic manufacturers would have made profits on only 1 out of 13 formulations of ARVs in both 2010 and 2012, and publicly owned manufacturers would have made profits on 5 and 3 out of 13 formulations in 2010 and 2012, respectively. We needed to assume a 20% and a 40% lower API cost for our model to predict that publicly owned and Indian manufacturers, respectively, would make profits on the sale of the majority of their ARVs. Between 2010 and 2012, we estimate that--across the ARV portfolio--the gross profit on sales of ARVs to low income countries decreased with between 6% and 7% of their sales price. Generic manufacturers consider that current prices are unsustainable. They suggested amendments to the tender procedures, simplified regulatory procedures, improved forecasting, and simplification of the ARV guidelines as critical improvements to maintain a viable ARV market. While recent price decreases indicate that there is still space for price reduction, our estimate that gross profit margin on sales decreased by 6 to 7% between 2010 and 2012 lends credibility to assertions by generic manufacturers that the ARV market in low income

  18. Antiretrovirals for low income countries: an analysis of the commercial viability of a highly competitive market

    Science.gov (United States)

    2013-01-01

    Background The price of antiretroviral drugs (ARVs) in low income countries declined steadily in recent years. This raises concerns about the commercial viability of the market of ARVs in low income countries. Methods Using 2 costing scenarios, we modeled the production cost of the most commonly used ARVs in low income countries in 2010 and 2012, and assessed whether, at the median price paid by low income countries, their manufacturers would still make profits. By interviews we consulted 11 generic manufacturers on the current state of the ARV market, and on what would be required to ensure their continued commitment to supply ARVs to low income countries. Results Using the lowest prices for active pharmaceutical ingredients (API) quoted to WHO, and applying published assumptions about the production cost of ARVs, our baseline estimate was that Indian generic manufacturers would have made profits on only 1 out of 13 formulations of ARVs in both 2010 and 2012, and publicly owned manufacturers would have made profits on 5 and 3 out of 13 formulations in 2010 and 2012, respectively. We needed to assume a 20% and a 40% lower API cost for our model to predict that publicly owned and Indian manufacturers, respectively, would make profits on the sale of the majority of their ARVs. Between 2010 and 2012, we estimate that - across the ARV portfolio - the gross profit on sales of ARVs to low income countries decreased with between 6% and 7% of their sales price. Generic manufacturers consider that current prices are unsustainable. They suggested amendments to the tender procedures, simplified regulatory procedures, improved forecasting, and simplification of the ARV guidelines as critical improvements to maintain a viable ARV market. Conclusions While recent price decreases indicate that there is still space for price reduction, our estimate that gross profit margin on sales decreased by 6 to 7% between 2010 and 2012 lends credibility to assertions by generic manufacturers

  19. Melatonin in Arabidopsis thaliana acts as plant growth regulator at low concentrations and preserves seed viability at high concentrations.

    Science.gov (United States)

    Hernández, Ismaél Gatica; Gomez, Federico José Vicente; Cerutti, Soledad; Arana, María Verónica; Silva, María Fernanda

    2015-09-01

    Since the discovery of melatonin in plants, several roles have been described for different species, organs, and developmental stages. Arabidopsis thaliana, being a model plant species, is adequate to contribute to the elucidation of the role of melatonin in plants. In this work, melatonin was monitored daily by UHPLC-MS/MS in leaves, in order to study its diurnal accumulation as well as the effects of natural and artificial light treatments on its concentration. Furthermore, the effects of exogenous application of melatonin to assess its role in seed viability after heat stress and as a regulator of growth and development of vegetative tissues were evaluated. Our results indicate that melatonin contents in Arabidopsis were higher in plants growing under natural radiation when compared to those growing under artificial conditions, and its levels were not diurnally-regulated. Exogenous melatonin applications prolonged seed viability after heat stress conditions. In addition, melatonin applications retarded leaf senescence. Its effects as growth promoter were dose and tissue-dependent; stimulating root growth at low concentrations and decreasing leaf area at high doses. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  20. Consortium inoculum of five thermo-tolerant phosphate solubilizing Actinomycetes for multipurpose biofertilizer preparation.

    Science.gov (United States)

    Nandimath, Arusha P; Karad, Dilip D; Gupta, Shantikumar G; Kharat, Arun S

    2017-10-01

    Alkaline pH of the soil facilitates the conversion of phosphate present in phosphate fertilizer applied in the field to insoluble phosphate which is not available to plants. Problem of soluble phosphate deficiency arises, primarily due to needless use of phosphate fertilizer. We sought to biofertilizer with the thermo-tolerant phosphate solubilizing actinomycetes consortium that could convert insoluble phosphate to soluble phosphate at wider temperature range. In the present investigation consortium of five thermo-tolerant phosphate solubilizing actinomycetes was applied for preparation of inoculum to produce multipurpose bio-fertilizer. Phosphates solubilizing thermo-tolerant 32 actinomycetes strains were processed for identification with the use of PIBWIN software and were screened for phosphate solubilizing activity. Amongst these five actinomycetes were selected on the basis of their ability to produce cellulase, chitinase, pectinase, protease, lipase, amylase and phosphate solubilizing enzymes. Ability to produce these enzymes at 28°C and 50°C were examined. Biofertilizer was prepared by using agricultural waste as a raw material. While preparation of bio-fertilizer the pH decreased from 7.5 to 4.3 and temperature increased up to 74°C maximum at the end of 4 th week and in subsequent week it started to decline gradually till it reached around 50°C, which was found to be stable up to eighth week. This thermo-tolerant actinomycetes consortium released soluble phosphate of up to 46.7 μg ml -1 . As the mesophilic organisms die out at high temperature of composting hence thormo-tolerant actinomycetes would be the better substitute for preparation of phosphate solubilizing bio-fertilizer with added potential to degrade complex macromolecules in composting.

  1. Screening of thermotolerant and thermophilic fungi aiming β-xylosidase and arabinanase production

    Science.gov (United States)

    Benassi, Vivian Machado; de Lucas, Rosymar Coutinho; Jorge, João Atílio; Polizeli, Maria de Lourdes Teixeira de Moraes

    2014-01-01

    Plant cell wall is mainly composed by cellulose, hemicellulose and lignin. The heterogeneous structure and composition of the hemicellulose are key impediments to its depolymerization and subsequent use in fermentation processes. Thus, this study aimed to perform a screening of thermophilic and thermotolerant filamentous fungi collected from different regions of the São Paulo state, and analyze the production of β-xylosidase and arabinanase at different temperatures. These enzymes are important to cell wall degradation and synthesis of end products as xylose and arabinose, respectively, which are significant sugars to fermentation and ethanol production. A total of 12 fungal species were analyzed and 9 of them grew at 45 °C, suggesting a thermophilic or thermotolerant character. Additionally Aspergillus thermomutatus anamorph of Neosartorya and A. parasiticus grew at 50 °C. Aspergillus niger and Aspergillus thermomutatus were the filamentous fungi with the most expressive production of β-xylosidase and arabinanase, respectively. In general for most of the tested microorganisms, β-xylosidase and arabinanase activities from mycelial extract (intracellular form) were higher in cultures grown at high temperatures (35–40 °C), while the correspondent extracellular activities were favorably secreted from cultures at 30 °C. This study contributes to catalogue isolated fungi of the state of São Paulo, and these findings could be promising sources for thermophilic and thermotolerant microorganisms, which are industrially important due to their enzymes. PMID:25763055

  2. Screening of thermotolerant and thermophilic fungi aiming β-xylosidase and arabinanase production

    Directory of Open Access Journals (Sweden)

    Vivian Machado Benassi

    2014-12-01

    Full Text Available Plant cell wall is mainly composed by cellulose, hemicellulose and lignin. The heterogeneous structure and composition of the hemicellulose are key impediments to its depolymerization and subsequent use in fermentation processes. Thus, this study aimed to perform a screening of thermophilic and thermotolerant filamentous fungi collected from different regions of the São Paulo state, and analyze the production of β-xylosidase and arabinanase at different temperatures. These enzymes are important to cell wall degradation and synthesis of end products as xylose and arabinose, respectively, which are significant sugars to fermentation and ethanol production. A total of 12 fungal species were analyzed and 9 of them grew at 45 ºC, suggesting a thermophilic or thermotolerant character. Additionally Aspergillus thermomutatus anamorph of Neosartorya and A. parasiticus grew at 50 ºC. Aspergillus niger and Aspergillus thermomutatus were the filamentous fungi with the most expressive production of β-xylosidase and arabinanase, respectively. In general for most of the tested microorganisms, β-xylosidase and arabinanase activities from mycelial extract (intracellular form were higher in cultures grown at high temperatures (35-40 ºC, while the correspondent extracellular activities were favorably secreted from cultures at 30 ºC. This study contributes to catalogue isolated fungi of the state of São Paulo, and these findings could be promising sources for thermophilic and thermotolerant microorganisms, which are industrially important due to their enzymes.

  3. Thermotolerant yeasts and application for ethanol production

    Directory of Open Access Journals (Sweden)

    To-on, N.

    2007-07-01

    Full Text Available A total of 70 thermotolerant yeast strains were isolated at 40oC from 145 samples including fruit, leaves, flowers, soils and oil-palm fruits. Six isolates showed maximum growth at 40oC within 18 h. Three isolates (MIY1, MIY48 and MIY57 were selected based on their ability to ferment glucose and sucrose rapidly (24 h and showed the maximum temperature for growth at 42oC but it was good at 40oC. MIY57 produced 4.6% (v/v ethanol at 40oC from a medium containing 15% glucose. The optimum cultivation conditions for growth and ethanol production of MIY57 was 5% inoculum into the fermentation medium containing 15% glucose and 1% yeast extract with initial pH of 4.5 on a shaking incubator at 150 rpm at 40oC. MIY57, under these conditions, produced maximum ethanol of 5.0% (v/v after 48 h incubation while S. cerevisiae TISTR 5048 produced only 3.7% (v/v. Maximum cell dry weight was 7.2 g/L (at 18 h, again much higher than that of S. cerevisiae TISTR 5048 (4.1 g/L. Based on morphological, physiological and molecular studies, this strain (MIY57 was identified as Saccharomyces cerevisiae.

  4. Classical and Molecular Identification of Thermotolerant Campylobacters from Poultry Meat

    Directory of Open Access Journals (Sweden)

    Tina Zorman

    2002-01-01

    Full Text Available Poultry meat samples from Slovenian retail market were examined for the presence of thermotolerant campylobacters. The isolates were identified by phenotypic and genotypic methods. ISO 10272 recommendations were followed for phenotypic identification. Different PCR assays, targeting species specific DNA regions in C. jejuni and C. coli, were checked for their applicability in identification. High degree of tested samples was positive (27/33, with significant proportion of C. coli (32 % among identified strains. High percentage of C. jejuni strains (54 % were hippurate negative. Phenotypic identification was therefore found to be inconvenient because of the presence of the strains with atypical phenotype and possible misinterpretation of test results. Multiplex PCR, targeting hippuricase gene in C. jejuni and species specific region in C. coli, was found to be an efficient method that allowed fast, simple and accurate identification of C. jejuni and C. coli. FlaA PCR is a reliable method to identify the group C. jejuni/C. coli, but it does not differentiate between the two species. CdtB PCR is inconvenient because of many false negative and some false positive results.

  5. Heat Stress Phenotypes of Arabidopsis Mutants Implicate Multiple Signaling Pathways in the Acquisition of Thermotolerance1[w

    Science.gov (United States)

    Larkindale, Jane; Hall, Jennifer D.; Knight, Marc R.; Vierling, Elizabeth

    2005-01-01

    To investigate the importance of different processes to heat stress tolerance, 45 Arabidopsis (Arabidopsis thaliana) mutants and one transgenic line were tested for basal and acquired thermotolerance at different stages of growth. Plants tested were defective in signaling pathways (abscisic acid, salicylic acid, ethylene, and oxidative burst signaling) and in reactive oxygen metabolism (ascorbic acid or glutathione production, catalase) or had previously been found to have temperature-related phenotypes (e.g. fatty acid desaturase mutants, uvh6). Mutants were assessed for thermotolerance defects in seed germination, hypocotyl elongation, root growth, and seedling survival. To assess oxidative damage and alterations in the heat shock response, thiobarbituric acid reactive substances, heat shock protein 101, and small heat shock protein levels were determined. Fifteen mutants showed significant phenotypes. Abscisic acid (ABA) signaling mutants (abi1 and abi2) and the UV-sensitive mutant, uvh6, showed the strongest defects in acquired thermotolerance of root growth and seedling survival. Mutations in nicotinamide adenine dinucleotide phosphate oxidase homolog genes (atrbohB and D), ABA biosynthesis mutants (aba1, aba2, and aba3), and NahG transgenic lines (salicylic acid deficient) showed weaker defects. Ethylene signaling mutants (ein2 and etr1) and reactive oxygen metabolism mutants (vtc1, vtc2, npq1, and cad2) were more defective in basal than acquired thermotolerance, especially under high light. All mutants accumulated wild-type levels of heat shock protein 101 and small heat shock proteins. These data indicate that, separate from heat shock protein induction, ABA, active oxygen species, and salicylic acid pathways are involved in acquired thermotolerance and that UVH6 plays a significant role in temperature responses in addition to its role in UV stress. PMID:15923322

  6. Influence of high gas production during thermophilic anaerobic digestion in pilot-scale and lab-scale reactors on survival of the thermotolerant pathogens Clostridium perfringens and Campylobacter jejuni in piggery wastewater.

    Science.gov (United States)

    Skillman, L C; Bajsa, O; Ho, L; Santhanam, B; Kumar, M; Ho, G

    2009-07-01

    Safe reuse of animal wastes to capture energy and nutrients, through anaerobic digestion processes, is becoming an increasingly desirable solution to environmental pollution. Pathogen decay is the most important safety consideration and is in general, improved at elevated temperatures and longer hydraulic residence times. During routine sampling to assess pathogen decay in thermophilic digestion, an inversely proportional relationship between levels of Clostridium perfringens and gas production was observed. Further samples were collected from pilot-scale, bench-scale thermophilic reactors and batch scale vials to assess whether gas production (predominantly methane) could be a useful indicator of decay of the thermotolerant pathogens C. perfringens and Campylobacter jejuni. Pathogen levels did appear to be lower where gas production and levels of methanogens were higher. This was evident at each operating temperature (50, 57, 65 degrees C) in the pilot-scale thermophilic digesters, although higher temperatures also reduced the numbers of pathogens detected. When methane production was higher, either when feed rate was increased, or pH was lowered from 8.2 (piggery wastewater) to 6.5, lower numbers of pathogens were detected. Although a number of related factors are known to influence the amount and rate of methane production, it may be a useful indicator of the removal of the pathogens C. perfringens and C. jejuni.

  7. Nutrient limitation and high irradiance acclimation reduce PAR and UV-induced viability loss in the Antarctic diatom Chaetoceros brevis (Bacillariophyceae)

    NARCIS (Netherlands)

    van de Poll, WH; van Leeuwe, MA; Roggeveld, J; Buma, AGJ

    The effects of high PAR (400-700 nm), UVA (315-400 nm), and UVB (280-315 nm) radiation on viability and photosynthesis were investigated for Chaetoceros brevis Schutt. This Antarctic marine diatom was cultivated under low, medium, and high irradiance and nitrate, phosphate, silicate, and iron

  8. Silk-fibronectin protein alloy fibres support cell adhesion and viability as a high strength, matrix fibre analogue

    Science.gov (United States)

    Jacobsen, Matthew M.; Li, David; Gyune Rim, Nae; Backman, Daniel; Smith, Michael L.; Wong, Joyce Y.

    2017-01-01

    Silk is a natural polymer with broad utility in biomedical applications because it exhibits general biocompatibility and high tensile material properties. While mechanical integrity is important for most biomaterial applications, proper function and integration also requires biomaterial incorporation into complex surrounding tissues for many physiologically relevant processes such as wound healing. In this study, we spin silk fibroin into a protein alloy fibre with whole fibronectin using wet spinning approaches in order to synergize their respective strength and cell interaction capabilities. Results demonstrate that silk fibroin alone is a poor adhesive surface for fibroblasts, endothelial cells, and vascular smooth muscle cells in the absence of serum. However, significantly improved cell attachment is observed to silk-fibronectin alloy fibres without serum present while not compromising the fibres’ mechanical integrity. Additionally, cell viability is improved up to six fold on alloy fibres when serum is present while migration and spreading generally increase as well. These findings demonstrate the utility of composite protein alloys as inexpensive and effective means to create durable, biologically active biomaterials. PMID:28378749

  9. Silk-fibronectin protein alloy fibres support cell adhesion and viability as a high strength, matrix fibre analogue

    Science.gov (United States)

    Jacobsen, Matthew M.; Li, David; Gyune Rim, Nae; Backman, Daniel; Smith, Michael L.; Wong, Joyce Y.

    2017-04-01

    Silk is a natural polymer with broad utility in biomedical applications because it exhibits general biocompatibility and high tensile material properties. While mechanical integrity is important for most biomaterial applications, proper function and integration also requires biomaterial incorporation into complex surrounding tissues for many physiologically relevant processes such as wound healing. In this study, we spin silk fibroin into a protein alloy fibre with whole fibronectin using wet spinning approaches in order to synergize their respective strength and cell interaction capabilities. Results demonstrate that silk fibroin alone is a poor adhesive surface for fibroblasts, endothelial cells, and vascular smooth muscle cells in the absence of serum. However, significantly improved cell attachment is observed to silk-fibronectin alloy fibres without serum present while not compromising the fibres’ mechanical integrity. Additionally, cell viability is improved up to six fold on alloy fibres when serum is present while migration and spreading generally increase as well. These findings demonstrate the utility of composite protein alloys as inexpensive and effective means to create durable, biologically active biomaterials.

  10. Inhibitory effects of multiwall carbon nanotubes with high iron impurity on viability and neuronal differentiation in cultured PC12 cells.

    Science.gov (United States)

    Meng, Li; Jiang, Aihua; Chen, Rui; Li, Chen-zhong; Wang, Liming; Qu, Ying; Wang, Peng; Zhao, Yuliang; Chen, Chunying

    2013-11-08

    The increasing use of carbon nanotubes (CNTs) in biomedical applications has garnered a great concern on their potential negative effects to human health. CNTs have been reported to potentially disrupt normal neuronal function and they were speculated to accumulate and cause brain damage, although a lot of distinct and exceptional properties and potential wide applications have been associated with this material in neurobiology. Fe impurities strapped inside the CNTs may be partially responsible for neurotoxicity generation. In the present study, we selected rat pheochromocytoma (PC12) cells to investigate and compare the effects of two kinds of multiwall carbon nanotubes (MWCNTs) with different concentrations of Fe impurities which usually come from the massive production of CNTs by chemical vapor deposition. Exposure to Fe-high MWCNTs can reduce cell viability and increase cytoskeletal disruption of undifferentiated PC12 cells, diminish the ability to form mature neurites, and then adversely influence the neuronal dopaminergic phenotype in NGF-treated PC-12 cells. The present results highlight the critical role of iron residue in the adverse response to MWCNTs exposure in neural cells. These findings provide useful information for understanding the toxicity and safe application of carbon nanotubes. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  11. The prevalence of thermotolerant Campylobacter species in food ...

    African Journals Online (AJOL)

    Background: Thermotolerant Campylobacter spp. is known to occur in the intestinal systems of a wide variety of domestic and wild animals. Although Campylobacter jejuni and Campylobacter coli cause acute diarrhoeal diseases in humans worldwide, they mostly manifest themselves in an apparently healthy carrier state in ...

  12. Polyphasic identification of a new thermotolerant species of lactic ...

    African Journals Online (AJOL)

    Two thermotolerant and desiccation tolerant lactic acid bacteria (TDLAB) were pointed out from twenty isolated strains from soils and dried chicken faeces. Samples were collected in poultry farms in the vicinity of Dakar, Senegal (West Africa). The two new isolates were called Sp.4 (Sp.4=CWBI-B534=LMG7278) and Sp.20 ...

  13. Environmental regulation of alcohol metabolism in thermotolerant methylotrophic Bacillus strains

    NARCIS (Netherlands)

    Arfman, N.; Moezelaar, H.R.; Attwood, M.M.; Robinson, G.K.; Geel, M. van; Dijkhuizen, L.

    The thermotolerant methylotroph Bacillus sp. C1 possesses a novel NAD-dependent methanol dehydrogenase (MDH), with distinct structural and mechanistic properties. During growth on methanol and ethanol, MDH was responsible for the oxidation of both these substrates. MDH activity in cells grown on

  14. The prevalence of thermotolerant Campylobacter species in food ...

    African Journals Online (AJOL)

    Preferred Customer

    Abstract. Background: Thermotolerant Campylobacter spp. is known to occur in the intestinal systems of a wide variety of domestic and wild animals. Although Campylobacter jejuni and Campylobacter coli cause acute diarrhoeal diseases in humans worldwide, they mostly manifest themselves in an apparently healthy ...

  15. Induction of thermotolerance through heat acclimation and salicylic ...

    African Journals Online (AJOL)

    Many putative signaling molecules like SA, ABA, H2O2 and CaCl2 and heat acclimation have been found to be involved in inducing thermotolerance as well as in initiating the underlying signal transduction pathway for upregulating various genes involved in thermotolerace. In the present investigation, we observed the ...

  16. Fever-range hyperthermia vs. hypothermia effect on cancer cell viability, proliferation and HSP90 expression.

    Science.gov (United States)

    Kalamida, Dimitra; Karagounis, Ilias V; Mitrakas, Achilleas; Kalamida, Sofia; Giatromanolaki, Alexandra; Koukourakis, Michael I

    2015-01-01

    The current study examines the effect of fever-range hyperthermia and mild hypothermia on human cancer cells focusing on cell viability, proliferation and HSP90 expression. A549 and H1299 lung carcinoma, MCF7 breast adenocarcinoma, U87MG and T98G glioblastoma, DU145 and PC3 prostate carcinoma and MRC5 normal fetal lung fibroblasts cell lines were studied. After 3-day exposure to 34°C, 37°C and 40°C, cell viability was determined. Cell proliferation (ki67 index), apoptosis (Caspase 9) and HSP90 expression was studied by confocal microscopy. Viability/proliferation experiments demonstrated that MRC5 fibroblasts were extremely sensitive to hyperthermia, while they were the most resistant to hypothermia. T98G and A549 were thermo-tolerant, the remaining being thermo-sensitive to a varying degree. Nonetheless, as a universal effect, hypothermia reduced viability/proliferation in all cell lines. Hyperthermia sharply induced Caspase 9 in the U87MG most thermo-sensitive cell line. In T98G and A549 thermo-tolerant cell lines, the levels of Caspase 9 declined. Moreover, hyperthermia strongly induced the HSP90 levels in T98G, whilst a sharp decrease was recorded in the thermo-sensitive PC3 and U87MG cell lines. Hyperthermia sensitized thermo-sensitive cancer cell lines to cisplatin and temozolomide, whilst its sensitizing effect was diminished in thermo-tolerant cell lines. The existence of thermo-tolerant and thermo-sensitive cancer cell lines was confirmed, which further encourages research to classify human tumor thermic predilection for patient stratification in clinical trials. Of interest, mild hypothermia had a universal suppressing effect on cancer cell proliferation, further supporting the radio-sensitization hypothesis through reduction of oxygen and metabolic demands.

  17. Fever-range hyperthermia vs. hypothermia effect on cancer cell viability, proliferation and HSP90 expression.

    Directory of Open Access Journals (Sweden)

    Dimitra Kalamida

    Full Text Available The current study examines the effect of fever-range hyperthermia and mild hypothermia on human cancer cells focusing on cell viability, proliferation and HSP90 expression.A549 and H1299 lung carcinoma, MCF7 breast adenocarcinoma, U87MG and T98G glioblastoma, DU145 and PC3 prostate carcinoma and MRC5 normal fetal lung fibroblasts cell lines were studied. After 3-day exposure to 34°C, 37°C and 40°C, cell viability was determined. Cell proliferation (ki67 index, apoptosis (Caspase 9 and HSP90 expression was studied by confocal microscopy.Viability/proliferation experiments demonstrated that MRC5 fibroblasts were extremely sensitive to hyperthermia, while they were the most resistant to hypothermia. T98G and A549 were thermo-tolerant, the remaining being thermo-sensitive to a varying degree. Nonetheless, as a universal effect, hypothermia reduced viability/proliferation in all cell lines. Hyperthermia sharply induced Caspase 9 in the U87MG most thermo-sensitive cell line. In T98G and A549 thermo-tolerant cell lines, the levels of Caspase 9 declined. Moreover, hyperthermia strongly induced the HSP90 levels in T98G, whilst a sharp decrease was recorded in the thermo-sensitive PC3 and U87MG cell lines. Hyperthermia sensitized thermo-sensitive cancer cell lines to cisplatin and temozolomide, whilst its sensitizing effect was diminished in thermo-tolerant cell lines.The existence of thermo-tolerant and thermo-sensitive cancer cell lines was confirmed, which further encourages research to classify human tumor thermic predilection for patient stratification in clinical trials. Of interest, mild hypothermia had a universal suppressing effect on cancer cell proliferation, further supporting the radio-sensitization hypothesis through reduction of oxygen and metabolic demands.

  18. Sporophyte and gametophyte generations differ in their thermotolerance response in the moss Microbryum.

    Science.gov (United States)

    McLetchie, D Nicholas; Stark, Lloyd R

    2006-04-01

    Actively growing post-embryonic sporophytes of desert mosses are restricted to the cooler, wetter months. However, most desert mosses have perennial gametophytes. It is hypothesized that these life history patterns are due in part to a reduced thermotolerance for sporophytes relative to gametophytes. Gametophytes with attached embryonic sporophytes of Microbryum starckeanum were exposed whilst desiccated to thermal episodes of 35 degrees C (1 hr), 55 degrees C (1 hr), 75 degrees C (1 hr) and 75 degrees C (3 hr), then moistened and allowed to recover for 35 d in a growth chamber. All of the gametophytes survived the thermal exposures and produced protonemata, with the majority also producing shoot buds. Symptoms of gametophytic stress (leaf burning and discoloration of entire shoots) were present in lower frequencies in the 55 degrees C exposure. Sporophyte resumption of growth and maturation to meiosis were significantly negatively affected by thermal treatment. Not a single sporophyte exposed to the two higher thermal treatments (75 degrees C for 1 h and 75 degrees C for 3 h) survived to meiosis, and those sporophytes exposed to 75 degrees C that survived to the post-embryonic phenophase took significantly longer to reach this phase. Furthermore, among the thermal treatments where some capsules reached maturity (35 degrees C and 55 degrees C), maternal shoots that produced a meiotic capsule took longer to regenerate through protonemata than maternal shoots aborting their sporophyte, suggestive of a resource trade-off between generations. Either (1) the inherent sporophyte thermotolerance is quite low even in this desert moss, and/or (2) a gametophytic thermal stress response controls sporophyte viability.

  19. Directed evolution of a filamentous fungus for thermotolerance.

    Science.gov (United States)

    de Crecy, Eudes; Jaronski, Stefan; Lyons, Benjamin; Lyons, Thomas J; Keyhani, Nemat O

    2009-08-26

    Filamentous fungi are the most widely used eukaryotic biocatalysts in industrial and chemical applications. Consequently, there is tremendous interest in methodology that can use the power of genetics to develop strains with improved performance. For example, Metarhizium anisopliae is a broad host range entomopathogenic fungus currently under intensive investigation as a biologically based alternative to chemical pesticides. However, it use is limited by the relatively low tolerance of this species to abiotic stresses such as heat, with most strains displaying little to no growth between 35-37 degrees C. In this study, we used a newly developed automated continuous culture method called the Evolugator, which takes advantage of a natural selection-adaptation strategy, to select for thermotolerant variants of M. anisopliae strain 2575 displaying robust growth at 37 degrees C. Over a 4 month time course, 22 cycles of growth and dilution were used to select 2 thermotolerant variants of M. anisopliae. Both variants displayed robust growth at 36.5 degrees C, whereas only one was able to grow at 37 degrees C. Insect bioassays using Melanoplus sanguinipes (grasshoppers) were also performed to determine if thermotolerant variants of M. anisopliae retained entomopathogenicity. Assays confirmed that thermotolerant variants were, indeed, entomopathogenic, albeit with complex alterations in virulence parameters such as lethal dose responses (LD50) and median survival times (ST50). We report the experimental evolution of a filamentous fungus via the novel application of a powerful new continuous culture device. This is the first example of using continuous culture to select for complex phenotypes such as thermotolerance. Temperature adapted variants of the insect-pathogenic, filamentous fungus M. anisopliae were isolated and demonstrated to show vigorous growth at a temperature that is inhibitory for the parent strain. Insect virulence assays confirmed that pathogenicity can be

  20. Thermotolerance of an inactivated rabies vaccine for dogs.

    Science.gov (United States)

    Lankester, Felix J; Wouters, Pieter A W M; Czupryna, Anna; Palmer, Guy H; Mzimbiri, Imam; Cleaveland, Sarah; Francis, Mike J; Sutton, David J; Sonnemans, Denny G P

    2016-11-04

    This study provides the first robust data that the antibody response of dogs vaccinated with Nobivac® Rabies vaccine stored for several months at high temperatures (up to 30°C) is not inferior to that of dogs vaccinated with vaccine stored under recommended cold-chain conditions (2-8°C). A controlled and randomized non-inferiority study was carried out comparing the four-week post vaccination serological responses of Tanzanian village dogs inoculated with vaccine which had been stored at elevated temperatures for different periods of time with those of dogs vaccinated with the same product stored according to label recommendations. Specifically, the neutralizing antibody response following the use of vaccine which had been stored for up to six months at 25°C or for three months at 30°C was not inferior to that following the use of cold-chain stored vaccine. These findings provide reassurance that the vaccine is likely to remain efficacious even if exposed to elevated temperatures for limited periods of time and, under these circumstances, it can safely be used and not necessarily destroyed or discarded. The availability of thermotolerant vaccines has been an important factor in the success of several disease control and elimination programs and could greatly increase the capacity of rabies vaccination campaigns to access hard to reach communities in Africa and Asia. We have not confirmed a 3-year duration of immunity for the high temperature stored vaccine, however because annual re-vaccination is usually practiced for dogs presented for vaccination during campaigns in Africa and Asia this should not be a cause for concern. These findings will provide confidence that, for rabies control and elimination programs using this vaccine in low-income settings, more flexible delivery models could be explored, including those that involve limited periods of transportation and storage at temperatures higher than that currently recommended. Copyright © 2016 The Authors

  1. Thermotolerance responses in ripening berries of Vitis vinifera L. cv Muscat Hamburg.

    Science.gov (United States)

    Carbonell-Bejerano, Pablo; Santa María, Eva; Torres-Pérez, Rafael; Royo, Carolina; Lijavetzky, Diego; Bravo, Gema; Aguirreolea, Jone; Sánchez-Díaz, Manuel; Antolín, M Carmen; Martínez-Zapater, José M

    2013-07-01

    Berry organoleptic properties are highly influenced by ripening environmental conditions. In this study, we used grapevine fruiting cuttings to follow berry ripening under different controlled conditions of temperature and irradiation intensity. Berries ripened at higher temperatures showed reduced anthocyanin accumulation and hastened ripening, leading to a characteristic drop in malic acid and total acidity. The GrapeGen GeneChip® combined with a newly developed GrapeGen 12Xv1 MapMan version were utilized for the functional analysis of berry transcriptomic differences after 2 week treatments from veraison onset. These analyses revealed the establishment of a thermotolerance response in berries under high temperatures marked by the induction of heat shock protein (HSP) chaperones and the repression of transmembrane transporter-encoding transcripts. The thermotolerance response was coincident with up-regulation of ERF subfamily transcription factors and increased ABA levels, suggesting their participation in the maintenance of the acclimation response. Lower expression of amino acid transporter-encoding transcripts at high temperature correlated with balanced amino acid content, suggesting a transcriptional compensation of temperature effects on protein and membrane stability to allow for completion of berry ripening. In contrast, the lower accumulation of anthocyanins and higher malate metabolization measured under high temperature might partly result from imbalance in the expression and function of their specific transmembrane transporters and expression changes in genes involved in their metabolic pathways. These results open up new views to improve our understanding of berry ripening under high temperatures.

  2. Increased viability and resilience of haemolymph cells in blue mussels following pre-treatment with acute high-dose gamma irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Jaeschke, B. [Stockholm University (Sweden)

    2014-07-01

    In an initial experiment, blue mussels (Mytilus edulis) were exposed to a range of acute high doses of gamma radiation in the laboratory. Haemolymph was extracted and the haemocytes (blood cells) were scored for cell viability (% living cells) under a microscope, directly after irradiation (0.04, 0.4 or 4 Gy) and again after a subsequent treatment with hydrogen peroxide in vitro (final H{sub 2}O{sub 2} conc.: 0.2 μM). Cell viability in controls (0 Gy) was approximately 100% and no cell death was observable from radiation exposure alone. When treated with H{sub 2}O{sub 2} a decrease in cell viability was seen across all treatments, however this decrease in viability was reduced with increasing radiation pre-treatment (0 Gy = 53%; 0.04 Gy = 66%; 0.4 Gy = 75%; 4 Gy = 83%). To investigate the mechanism for this therapeutic effect observed, the experiment was repeated. Using mussels from a different location, the same, but more extensive method of irradiation (0[control], 0.04, 0.4 Gy, 5 or 40 Gy) and H{sub 2}O{sub 2} treatment was used. Additional haemolymph sub-samples were taken for analysis of catalase concentration. In this second experiment, viability of cells from controls was only 62%, indicating the mussels were in a poorer condition than those of the previous experiment. The lowest level of radiation exposure (0.04 Gy) further decreased the viability (56%). However, at higher doses the viability was increased compared to control, which then gradually declined with increasing dose (0.4 Gy = 75%; 5 Gy = 72%; 40 Gy = 65%). Catalase analysis demonstrated a complimentary pattern of activity of the antioxidant in the haemolymph, directly correlating with radiation dose (0 Gy = 0.2 U; 0.04 Gy = 0.1 U; 0.4 Gy = 1.3 U; 5 Gy = 0.9 U; 40 Gy = 0.1 Gy). Treatment with H{sub 2}O{sub 2} decreased cell viability across all treatments, but no pattern between radiation treatments was discernable. The results indicate that an acute dose of radiation not only has negligible

  3. Draft Genome Sequence of Natranaerobius trueperi DSM 18760T, an Anaerobic, Halophilic, Alkaliphilic, Thermotolerant Bacterium Isolated from a Soda Lake

    OpenAIRE

    Guo, Xiaomeng; Liao, Ziya; Holtzapple, Mark; Hu, Qingping; Zhao, Baisuo

    2017-01-01

    ABSTRACT The anaerobic, halophilic, alkaliphilic, thermotolerant bacterium Natranaerobius trueperi was isolated from a soda lake in Wadi An Natrun, Egypt. It grows optimally at 3.7?M Na+, pH?9.5, and 43?C. The draft genome consists of 2.63?Mb and is composed of 2,681 predicted genes. Genomic analysis showed that various genes are potentially involved in the adaptation mechanisms for osmotic stress, pH homeostasis, and high temperatures.

  4. Effect of UV radiation on thermotolerance, ethanol tolerance and osmotolerance of Saccharomyces cerevisiae VS1 and VS3 strains.

    Science.gov (United States)

    Sridhar, M; Sree, N Kiran; Rao, L Venkateswar

    2002-07-01

    After a previous mass screening and enrichment programme for the isolation of thermotolerant yeasts, VS1, VS2, VS3 and VS4 strains isolated from soil samples, collected within the hot regions of Kothagudem Thermal Power Plant, AP, India, had a better thermotolerance, osmotolerance and ethanol tolerance than the other isolates. Among these isolates VS1 and VS3 were best performers. Efforts were made to further improve their osmotolerance, thermotolerance and ethanol tolerance by treating them with UV radiation. Mutants of VS1 and VS3 produced more biomass and ethanol than the parent strains at high temperature and glucose concentrations. The amount of biomass produced by VS1 and VS3 mutants was 0.25 and 0.20 g l(-1) more than the parent strains at 42 degrees C using 2% glucose. At high glucose concentrations VS1 and VS3 mutants produced biomass which was 0.70 and 0.30 g l(-1) at 30 degrees C and 0.10 and 0.20 g l(-1) at 40 degrees C more than the parent strains. The amount of ethanol produced by the mutants (VS1 and VS3) was 8.20 and 1.20 g l(-1) more than the parent strains at 42 degrees C using 150 g l(-1) glucose. More ethanol was produced by mutants (VS1 and VS3) than the parents at high glucose concentrations of 5.0 and 6.0 g l(-1) at 30 degrees C and 13.0 and 3.0 g l(-1) at 42 degrees C, respectively. These results indicated that UV mutagenesis can be used for improving thermotolerance, ethanol tolerance and osmotolerance in VS1 and VS3 yeast strains.

  5. A review of acquired thermotolerance, heat shock proteins, and molecular chaperones in archaea

    Energy Technology Data Exchange (ETDEWEB)

    Trent, J.D.

    1996-05-01

    Acquired thermotolerance, the associated synthesis of heat-shock proteins (HSPs) under stress conditions, and the role of HSPs as molecular chaperones under normal growth conditions have been studied extensively in eukaryotes and bacteria, whereas research in these areas in archaea is only beginning. All organisms have evolved a variety of strategies for coping with high-temperature stress, and among these strategies is the increased synthesis of HSPs. The facts that both high temperatures and chemical stresses induce the HSPs and that some of the HSPs recognize and bind to unfolded proteins in vitro have led to the theory that the function of HSPs is to prevent protein aggregation in vivo. The facts that some HSPs are abundant under normal growth conditions and that they assist in protein folding in vitro have led to the theory that they assist protein folding in vivo; in this role, they are referred to as molecular chaperones. The limited research on acquired thermotolerance, HSPs, and molecular chaperones in archaea, particularly the hyperthermophilic archaea, suggests that these extremophiles provide a new perspective in these areas of research, both because they are members of a separate phylogenetic domain and because they have evolved to live under extreme conditions.

  6. Morphological and enzymatic response of the thermotolerant fungus Fomes sp. EUM1 in solid state fermentation under thermal stress.

    Science.gov (United States)

    Ordaz-Hernández, Armando; Ortega-Sánchez, Eric; Montesinos-Matías, Roberto; Hernández-Martínez, Ricardo; Torres-Martínez, Daniel; Loera, Octavio

    2016-08-01

    Thermotolerance of the fungus Fomes sp. EUM1 was evaluated in solid state fermentation (SSF). This thermotolerant strain improved both hyphal invasiveness (38%) and length (17%) in adverse thermal conditions exceeding 30°C and to a maximum of 40°C. In contrast, hyphal branching decreased by 46% at 45°C. The production of cellulases over corn stover increased 1.6-fold in 30°C culture conditions, xylanases increased 2.8-fold at 40°C, while laccase production improved 2.7-fold at 35°C. Maximum production of lignocellulolytic enzymes was obtained at elevated temperatures in shorter fermentation times (8-6 days), although the proteases appeared as a thermal stress response associated with a drop in lignocellulolytic activities. Novel and multiple isoenzymes of xylanase (four bands) and cellulase (six bands) were secreted in the range of 20-150 kDa during growth in adverse temperature conditions. However, only a single laccase isoenzyme (46 kDa) was detected. This is the first report describing the advantages of a thermotolerant white-rot fungus in SSF. These results have important implications for large-scale SSF, where effects of metabolic heat are detrimental to growth and enzyme production, which are severely affected by the formation of high temperature gradients. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. Prevalence of thermotolerant Campylobacter spp. in farmed hares (Lepus europaeus).

    Science.gov (United States)

    Santaniello, Antonio; Dipineto, Ludovico; Veneziano, Vincenzo; Mariani, Ugo; Fioretti, Alessandro; Menna, Lucia Francesca

    2014-10-01

    Thermotolerant Campylobacter spp. were isolated from 118/240 (49.2%) rectal swabs from commercially farmed hares (Lepus europaeus) in southern Italy. Using multiplex PCR, Campylobacter coli was identified in 118/118 (100%) positive samples, while 17/118 (14.4%) positive samples were also positive for Campylobacter jejuni. Adult hares had a higher prevalence of infection with Campylobacter spp. than juvenile hares. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Expression and thermotolerance of calreticulin during pollen development in tobacco

    Czech Academy of Sciences Publication Activity Database

    Hrubá, Petra; Honys, David; Tupý, Jaroslav

    2005-01-01

    Roč. 18, č. 3 (2005), s. 143-148 ISSN 0934-0882 R&D Projects: GA ČR GP522/02/D075; GA MŠk LZ1K03018; GA AV ČR KJB6038409 Institutional research plan: CEZ:AV0Z50380511 Keywords : tobacco * pollen development * thermotolerance Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.278, year: 2005

  9. Salinity modulates thermotolerance, energy metabolism and stress response in amphipods Gammarus lacustris.

    Science.gov (United States)

    Vereshchagina, Kseniya P; Lubyaga, Yulia A; Shatilina, Zhanna; Bedulina, Daria; Gurkov, Anton; Axenov-Gribanov, Denis V; Baduev, Boris; Kondrateva, Elizaveta S; Gubanov, Mikhail; Zadereev, Egor; Sokolova, Inna; Timofeyev, Maxim

    2016-01-01

    Temperature and salinity are important abiotic factors for aquatic invertebrates. We investigated the influence of different salinity regimes on thermotolerance, energy metabolism and cellular stress defense mechanisms in amphipods Gammarus lacustris Sars from two populations. We exposed amphipods to different thermal scenarios and determined their survival as well as activity of major antioxidant enzymes (peroxidase, catalase, glutathione S-transferase) and parameters of energy metabolism (content of glucose, glycogen, ATP, ADP, AMP and lactate). Amphipods from a freshwater population were more sensitive to the thermal challenge, showing higher mortality during acute and gradual temperature change compared to their counterparts from a saline lake. A more thermotolerant population from a saline lake had high activity of antioxidant enzymes. The energy limitations of the freshwater population (indicated by low baseline glucose levels, downward shift of the critical temperature of aerobic metabolism and inability to maintain steady-state ATP levels during warming) was observed, possibly reflecting a trade-off between the energy demands for osmoregulation under the hypo-osmotic condition of a freshwater environment and protection against temperature stress.

  10. Laccase produced by a thermotolerant strain of Trametes trogii LK13.

    Science.gov (United States)

    Yan, Jinping; Chen, Yuhui; Niu, Jiezhen; Chen, Daidi; Chagan, Irbis

    2015-03-01

    Thermophilic and thermotolerant micro-organisms strains have served as the natural source of industrially relevant and thermostable enzymes. Although some strains of the Trametes genus are thermotolerant, few Trametes strains were studied at the temperature above 30 °C until now. In this paper, the laccase activity and the mycelial growth rate for Trametes trogii LK13 are superior at 37 °C. Thermostability and organic cosolvent tolerance assays of the laccase produced at 37 °C indicated that the enzyme possessed fair thermostability with 50% of its initial activity at 80 °C for 5 min, and could remain 50% enzyme activity treated with organic cosolvent at the concentration range of 25%-50% (v/v). Furthermore, the test on production of laccase and lignocellulolytic enzymes showed the crude enzymes possessed high laccase level (1000 U g (-1) ) along with low cellulose (2 U g (-1) ) and xylanase (140 U g (-1) ) activity. Thus, T. trogii LK13 is a potential strain to be applied in many biotechnological processes.

  11. Laccase produced by a thermotolerant strain of Trametes trogii LK13

    Directory of Open Access Journals (Sweden)

    Jinping Yan

    2015-03-01

    Full Text Available Thermophilic and thermotolerant micro-organisms strains have served as the natural source of industrially relevant and thermostable enzymes. Although some strains of the Trametes genus are thermotolerant, few Trametes strains were studied at the temperature above 30 °C until now. In this paper, the laccase activity and the mycelial growth rate for Trametes trogii LK13 are superior at 37 °C. Thermostability and organic cosolvent tolerance assays of the laccase produced at 37 °C indicated that the enzyme possessed fair thermostability with 50% of its initial activity at 80 °C for 5 min, and could remain 50% enzyme activity treated with organic cosolvent at the concentration range of 25%–50% (v/v. Furthermore, the test on production of laccase and lignocellulolytic enzymes showed the crude enzymes possessed high laccase level (1000 U g−1 along with low cellulose (2 U g−1 and xylanase (140 U g−1 activity. Thus, T. trogii LK13 is a potential strain to be applied in many biotechnological processes.

  12. Salinity modulates thermotolerance, energy metabolism and stress response in amphipods Gammarus lacustris

    Science.gov (United States)

    Vereshchagina, Kseniya P.; Lubyaga, Yulia A.; Shatilina, Zhanna; Bedulina, Daria; Gurkov, Anton; Axenov-Gribanov, Denis V.; Baduev, Boris; Kondrateva, Elizaveta S.; Gubanov, Mikhail; Zadereev, Egor; Sokolova, Inna

    2016-01-01

    Temperature and salinity are important abiotic factors for aquatic invertebrates. We investigated the influence of different salinity regimes on thermotolerance, energy metabolism and cellular stress defense mechanisms in amphipods Gammarus lacustris Sars from two populations. We exposed amphipods to different thermal scenarios and determined their survival as well as activity of major antioxidant enzymes (peroxidase, catalase, glutathione S-transferase) and parameters of energy metabolism (content of glucose, glycogen, ATP, ADP, AMP and lactate). Amphipods from a freshwater population were more sensitive to the thermal challenge, showing higher mortality during acute and gradual temperature change compared to their counterparts from a saline lake. A more thermotolerant population from a saline lake had high activity of antioxidant enzymes. The energy limitations of the freshwater population (indicated by low baseline glucose levels, downward shift of the critical temperature of aerobic metabolism and inability to maintain steady-state ATP levels during warming) was observed, possibly reflecting a trade-off between the energy demands for osmoregulation under the hypo-osmotic condition of a freshwater environment and protection against temperature stress. PMID:27896024

  13. Genome shuffling improves thermotolerance and glutamic acid production of Corynebacteria glutamicum.

    Science.gov (United States)

    Zheng, Pu; Liu, Miao; Liu, Xiao-de; Du, Qiao-Yan; Ni, Ye; Sun, Zhi-Hao

    2012-03-01

    Genome shuffling was used to improve the thermotolerance of L: -glutamic acid-producing strain Corynebacteria glutamicum. Five strains with subtle improvements in high temperature tolerance and productivity were selected by ultraviolet irradiation and diethyl sulfate mutagenesis. An improved strain (F343) was obtained by three rounds of genome shuffling of the five strains as mentioned above. The cell density of F343 was four times higher than that of ancestor strains after 24 h of cultivation at 44°C, and importantly, the yield of L: -glutamic acid was increased by 1.8-times comparing with that of the ancestor strain at 38°C in a 5-L fermentor. With glucose supplement and two-stage pH control, the L: -glutamate acid concentration of F343 reached 119 g/L after fermentation for 30 h. The genetic diversity between F343 and its ancestors was also evaluated by amplified fragment length polymorphism analysis. Results suggest that the phenotypes for both thermotolerance and L: -glutamic acid production in F343 were evolved.

  14. Salinity modulates thermotolerance, energy metabolism and stress response in amphipods Gammarus lacustris

    Directory of Open Access Journals (Sweden)

    Kseniya P. Vereshchagina

    2016-11-01

    Full Text Available Temperature and salinity are important abiotic factors for aquatic invertebrates. We investigated the influence of different salinity regimes on thermotolerance, energy metabolism and cellular stress defense mechanisms in amphipods Gammarus lacustris Sars from two populations. We exposed amphipods to different thermal scenarios and determined their survival as well as activity of major antioxidant enzymes (peroxidase, catalase, glutathione S-transferase and parameters of energy metabolism (content of glucose, glycogen, ATP, ADP, AMP and lactate. Amphipods from a freshwater population were more sensitive to the thermal challenge, showing higher mortality during acute and gradual temperature change compared to their counterparts from a saline lake. A more thermotolerant population from a saline lake had high activity of antioxidant enzymes. The energy limitations of the freshwater population (indicated by low baseline glucose levels, downward shift of the critical temperature of aerobic metabolism and inability to maintain steady-state ATP levels during warming was observed, possibly reflecting a trade-off between the energy demands for osmoregulation under the hypo-osmotic condition of a freshwater environment and protection against temperature stress.

  15. Directed evolution of a filamentous fungus for thermotolerance

    Directory of Open Access Journals (Sweden)

    Lyons Thomas J

    2009-08-01

    Full Text Available Abstract Background Filamentous fungi are the most widely used eukaryotic biocatalysts in industrial and chemical applications. Consequently, there is tremendous interest in methodology that can use the power of genetics to develop strains with improved performance. For example, Metarhizium anisopliae is a broad host range entomopathogenic fungus currently under intensive investigation as a biologically based alternative to chemical pesticides. However, it use is limited by the relatively low tolerance of this species to abiotic stresses such as heat, with most strains displaying little to no growth between 35–37°C. In this study, we used a newly developed automated continuous culture method called the Evolugator™, which takes advantage of a natural selection-adaptation strategy, to select for thermotolerant variants of M. anisopliae strain 2575 displaying robust growth at 37°C. Results Over a 4 month time course, 22 cycles of growth and dilution were used to select 2 thermotolerant variants of M. anisopliae. Both variants displayed robust growth at 36.5°C, whereas only one was able to grow at 37°C. Insect bioassays using Melanoplus sanguinipes (grasshoppers were also performed to determine if thermotolerant variants of M. anisopliae retained entomopathogenicity. Assays confirmed that thermotolerant variants were, indeed, entomopathogenic, albeit with complex alterations in virulence parameters such as lethal dose responses (LD50 and median survival times (ST50. Conclusion We report the experimental evolution of a filamentous fungus via the novel application of a powerful new continuous culture device. This is the first example of using continuous culture to select for complex phenotypes such as thermotolerance. Temperature adapted variants of the insect-pathogenic, filamentous fungus M. anisopliae were isolated and demonstrated to show vigorous growth at a temperature that is inhibitory for the parent strain. Insect virulence assays

  16. Rational synthetic combination genetic devices boosting high temperature ethanol fermentation

    Directory of Open Access Journals (Sweden)

    Huan Sun

    2017-06-01

    Full Text Available The growth and production of yeast in the industrial fermentation are seriously restrained by heat stress and exacerbated by heat induced oxidative stress. In this study, a novel synthetic biology approach was developed to globally boost the viability and production ability of S. cerevisiae at high temperature through rationally designing and combing heat shock protein (HSP and superoxide dismutase (SOD genetic devices to ultimately synergistically alleviate both heat stress and oxidative stress. HSP and SOD from extremophiles were constructed to be different genetic devices and they were preliminary screened by heat resistant experiments and anti-oxidative experiments, respectively. Then in order to customize and further improve thermotolerance of S. cerevisiae, the HSP genetic device and SOD genetic device were rationally combined. The results show the simply assemble of the same function genetic devices to solve heat stress or oxidative stress could not enhance the thermotolerance considerably. Only S. cerevisiae with the combination genetic device (FBA1p-sod-MB4-FBA1p-shsp-HB8 solving both stress showed 250% better thermotolerance than the control and displayed further 55% enhanced cell density compared with the strains with single FBA1p-sod-MB4 or FBA1p-shsp-HB8 at 42 °C. Then the most excellent combination genetic device was introduced into lab S. cerevisiae and industrial S. cerevisiae for ethanol fermentation. The ethanol yields of the two strains were increased by 20.6% and 26.3% compared with the control under high temperature, respectively. These results indicate synergistically defensing both heat stress and oxidative stress is absolutely necessary to enhance the thermotolerance and production of S. cerevisiae.

  17. High Nutrient Levels and TORC1 Activity Reduce Cell Viability following Prolonged Telomere Dysfunction and Cell Cycle Arrest

    Directory of Open Access Journals (Sweden)

    Julia Klermund

    2014-10-01

    Full Text Available Cells challenged with DNA damage activate checkpoints to arrest the cell cycle and allow time for repair. Successful repair coupled to subsequent checkpoint inactivation is referred to as recovery. When DNA damage cannot be repaired, a choice between permanent arrest and cycling in the presence of damage (checkpoint adaptation must be made. While permanent arrest jeopardizes future lineages, continued proliferation is associated with the risk of genome instability. We demonstrate that nutritional signaling through target of rapamycin complex 1 (TORC1 influences the outcome of this decision. Rapamycin-mediated TORC1 inhibition prevents checkpoint adaptation via both Cdc5 inactivation and autophagy induction. Preventing adaptation results in increased cell viability and hence proliferative potential. In accordance, the ability of rapamycin to increase longevity is dependent upon the DNA damage checkpoint. The crosstalk between TORC1 and the DNA damage checkpoint may have important implications in terms of therapeutic alternatives for diseases associated with genome instability.

  18. Survey of thermotolerant Campylobacter in pheasant (Phasianus colchicus

    Directory of Open Access Journals (Sweden)

    Alessandro Fioretti

    2010-01-01

    Full Text Available This study was conducted with the aim to evaluate the prevalence of thermotolerant Campylobacter spp. in pheasant (Phasianus colchicus in Southern Italy. To achieve this goal, 60 cloacal swabs were collected. Campylobacter spp. were isolated from 52 out of 60 cloacal swabs tested. As proved by PCR, 100% of the strains were identified as C. coli (52/52, and 10 out of the 52 (19.2% positive samples were also positive to C. jejuni. The pheasant, can be considered, at least theoretically, as potential Campylobacter carriers.

  19. High modulus biodegradable polyurethanes for applications in cardiovascular stents: Evaluation of in-vitro degradation and cell viability

    Directory of Open Access Journals (Sweden)

    Melissa eSagarito

    2015-05-01

    Full Text Available We have recently reported the mechanical properties and hydrolytic degradation behaviour of a series of NovoSorb™ [1] biodegradable polyurethanes (PUs prepared by varying the hard segment (HS weight percentage from 60-100. In this study the in-vitro degradation behaviour of the PUs with and without extracellular matrix (ECM coating was investigated under accelerated hydrolytic degradation (phosphate buffer saline; PBS/70°C conditions to allow complete degradation. The mass loss at different time intervals and the effect of aqueous degradation products on the viability and growth of Human Umbilical Vein Endothelial Cells (HUVEC were examined.The results showed that for most PUs in the series the degradation medium turned acidic and the extent of this was dependent on the HS percentage. As the HS decreased, a drop in pH was observed, suggesting that the by-products of soft segment (SS degradation causing the solution to be acidic. Most of the samples were completely eroded by 18 weeks, except PU containing more than 70% HS which only showed partial degradation with no significant change in appearance. The cytotoxicity tests on HUVEC cells of the aqueous extracts obtained after 18 weeks incubation showed that toxicity was dependent on 3 factors: dose, percentage of HS and degradation time. HUVEC growth is similar to but not identical to that observed with tissue culture polystyrene (TCPS standard. The results from this in-vitro cytotoxicity study suggest that the mixture of degradation products formed during the accelerated degradation of this PU series are not toxic to cells under the experimental conditions used.

  20. Subendocardial Viability Ratio Is Impaired in Highly Proteinuric Chronic Kidney Disease Patients With Low Estimated Glomerular Filtration Rate.

    Science.gov (United States)

    Ekart, Robert; Šegula, Anja; Hartman, Tanja; Hojs, Nina; Hojs, Radovan

    2016-06-01

    Proteinuria and estimated glomerular filtration rate (eGFR) are markers of chronic kidney disease (CKD) and cardiovascular disease. With applanation tonometry, pulse wave analysis and many hemodynamic data are available. One of them is the subendocardial viability ratio (SEVR) which represents a non-invasive measure of myocardial perfusion related to the work of the heart. The aim of our study was to investigate the importance of SEVR in proteinuric CKD patients and healthy subjects. We performed a cross-sectional study in a cohort of 90 non-dialysis CKD patients and 39 healthy controls. SEVR was assessed by radial applanation tonometry (SphygmoCor, Atcor, Australia). Blood samples and urine albumin-to-creatinine ratio (UACR) were analyzed. CKD patients were divided in four groups according to the UACR and eGFR: CKD group 1: UACR > 1000 mg/g and eGFR  1000 mg/g and eGFR >30 mL/min; CKD group 3: UACR 30 mL/min. Using one-way ANOVA, we found a statistically significant difference in SEVR only between CKD group 1 and all other CKD groups and healthy control group (P < 0.022). Results of our study show that only CKD patients with UACR more than 1000 mg/g and eGFR below 30 mL/min have significantly lower SEVR. © 2016 International Society for Apheresis, Japanese Society for Apheresis, and Japanese Society for Dialysis Therapy.

  1. Evaluation of resistance development and viability recovery by toxigenic and non-toxigenic Staphylococcus aureus strains after repeated cycles of high hydrostatic pressure.

    Science.gov (United States)

    Baptista, Inês; Queirós, Rui P; Cunha, Angela; Rocha, Sílvia M; Saraiva, Jorge A; Almeida, Adelaide

    2015-04-01

    In this work, the development of resistance and the recovery of growth after several consecutive cycles of high hydrostatic pressure (HPP) were for the first time evaluated in different strains of Staphylococcus aureus. Three strains of this important and highly resilient to HPP foodborne pathogen were used: a non-enterotoxigenic ATCC 6538 strain, treated with 600 MPa for 30 min at 20 °C, and the toxigenic strains 2153 MA (with enterotoxin A) and 2065 MA (with the enterotoxins A, G and I), treated with 600 MPa for 15 min at 20 °C. After the first treatment, surviving colonies were used to produce new bacterial cultures. This procedure was repeated nine times more for each bacterium or until total inactivation occurred. The inactivation profile of non-enterotoxic strain and the two enterotoxic strains did not change after consecutive cycles, but the toxic strain with three enterotoxins was completely inactivated after the fourth cycle. The three strains did not recover their viability after 14 days. The results indicate that HPP effectively inactivates non-toxigenic and toxigenic strains of S. aureus after a single treatment. The surviving bacteria did not develop resistance after 10 cycles of pressurization and did not recover their viability after 14 days of incubation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. Vascular Graft Impregnation with Antibiotics: The Influence of High Concentrations of Rifampin, Vancomycin, Daptomycin, and Bacteriophage Endolysin HY-133 on Viability of Vascular Cells.

    Science.gov (United States)

    Herten, Monika; Idelevich, Evgeny A; Sielker, Sonja; Becker, Karsten; Scherzinger, Anna S; Osada, Nani; Torsello, Giovanni B; Bisdas, Theodosios

    2017-06-27

    BACKGROUND Rifampin-soaked synthetic prosthetic grafts have been widely used for prevention or treatment of vascular graft infections (VGIs). This in vitro study investigated the effect of the antibiotics daptomycin and vancomycin and the new recombinant bacteriophage endolysin HY-133 on vascular cells, as potential alternatives compared to rifampin. MATERIAL AND METHODS Primary human ECs, vascular smooth muscle cells (vSMC), and fibroblasts were cultivated in 96-well plates and incubated with rifampin, daptomycin, vancomycin, and endolysin HY-133 for 24 h. Subsequently, after washing, cell viability was determined by measuring mitochondrial ATP concentration. Antibiotics were used in their corresponding minimum and maximum serum concentrations, in decimal multiples and in maximum soaking concentration. The experiments were performed in triplicate. RESULTS The 10-fold max serum concentrations of rifampin, daptomycin, and vancomycin did not influence viability of EC and vSMC (100 µg/ml, p>0.170). Higher concentrations of rifampin (>1 mg/ml) significantly (pEndolysin did not display any cytotoxicity towards vascular cells. CONCLUSIONS Results of this in vitro study show the high cytotoxicity of rifampin against vascular cells, and may re-initiate the discussion about the benefit of prophylactic pre-soaking in high concentrations of rifampin. Further studies are necessary to determine the influence of rifampin on the restoration of vessel functionality versus its prophylactic effect against VGIs. Future use of recombinant phage endolysins for alternative prophylactic strategies needs further investigations.

  3. Lantana camara for fuel ethanol production using thermotolerant yeast.

    Science.gov (United States)

    Pasha, C; Nagavalli, M; Rao, L Venkateswar

    2007-06-01

    Evaluation of Lantana camara's use as feedstock for fuel ethanol production. Lantana camara plant material was hydrolysed with 1% sulfuric acid for 18 h at room temperature, followed by heat treatment of 121 degrees C for 20 min. Hemicellulosic hydrolyzate was separated and used for detoxification by ethyl acetate and overliming. Cellulosic fraction was hydrolysed with Aspergillus niger crude cellulase enzyme for 18 h at 55 degrees C. Using 15% (dw/v) substrate 73 g l(-1) total reducing sugars were obtained to give 78.7% hydrolysis of carbohydrate content. Acid and enzyme hydrolyzates were mixed equally and used for fermentation with thermotolerant Saccharomyces cerevisiae (VS(3)). Yeast fermented L. camara hydrolyzate well with a fermentation efficiency of 83.7% to give an ethanol yield of 0.431 +/- 0.018 g ethanol pre g sugar and productivity of 0.5 +/- 0.021 g l(-1) h(-1). Even though inhibitors were present in L. camara hydrolyzate, maximum sugars were utilized by thermotolerant yeast. Use of L. camara for fuel ethanol production with improved strains and detoxification can be recommended.

  4. The viability of perilabyrinthine osteocytes

    DEFF Research Database (Denmark)

    Bloch, Sune Land; Kristensen, Søren Lund; Sørensen, Mads Sølvsten

    2012-01-01

    Bone remodeling is highly inhibited around the inner ear space, most likely by the anti-resorptive action of the inner ear cytokine osteoprotegerin (OPG) entering perilabyrinthine bone through the lacuno-canalicular porosity (LCP). This extracellular signaling pathway depends on the viability...

  5. Pichia pastoris Exhibits High Viability and a Low Maintenance Energy Requirement at Near-Zero Specific Growth Rates.

    Science.gov (United States)

    Rebnegger, Corinna; Vos, Tim; Graf, Alexandra B; Valli, Minoska; Pronk, Jack T; Daran-Lapujade, Pascale; Mattanovich, Diethard

    2016-08-01

    The yeast Pichia pastoris is a widely used host for recombinant protein production. Understanding its physiology at extremely low growth rates is a first step in the direction of decoupling product formation from cellular growth and therefore of biotechnological relevance. Retentostat cultivation is an excellent tool for studying microbes at extremely low specific growth rates but has so far not been implemented for P. pastoris Retentostat feeding regimes were based on the maintenance energy requirement (mS) and maximum biomass yield on glucose (YX /S (max)) estimated from steady-state glucose-limited chemostat cultures. Aerobic retentostat cultivation enabled reproducible, smooth transitions from a specific growth rate (μ) of 0.025 h(-1) to near-zero specific growth rates (μ growth rates, viability remained at least 97%. The value of mS at near-zero growth rates was 3.1 ± 0.1 mg glucose per g biomass and h, which was 3-fold lower than the mS estimated from faster-growing chemostat cultures. This difference indicated that P. pastoris reduces its maintenance energy requirement at extremely low μ, a phenomenon not previously observed in eukaryotes. Intracellular levels of glycogen and trehalose increased, while μ progressively declined during retentostat cultivation. Transcriptional reprogramming toward zero growth included the upregulation of many transcription factors as well as stress-related genes and the downregulation of cell cycle genes. This study underlines the relevance of comparative analysis of maintenance energy metabolism, which has an important impact on large-scale industrial processes. The yeast Pichia pastoris naturally lives on trees and can utilize different carbon sources, among them glucose, glycerol, and methanol. In biotechnology, it is widely used for the production of recombinant proteins. For both the understanding of life in its natural habitat and optimized production processes, a better understanding of cell physiology at an

  6. Draft Genome Sequence of Natranaerobius trueperi DSM 18760T, an Anaerobic, Halophilic, Alkaliphilic, Thermotolerant Bacterium Isolated from a Soda Lake.

    Science.gov (United States)

    Guo, Xiaomeng; Liao, Ziya; Holtzapple, Mark; Hu, Qingping; Zhao, Baisuo

    2017-09-07

    The anaerobic, halophilic, alkaliphilic, thermotolerant bacterium Natranaerobius trueperi was isolated from a soda lake in Wadi An Natrun, Egypt. It grows optimally at 3.7 M Na+, pH 9.5, and 43°C. The draft genome consists of 2.63 Mb and is composed of 2,681 predicted genes. Genomic analysis showed that various genes are potentially involved in the adaptation mechanisms for osmotic stress, pH homeostasis, and high temperatures. Copyright © 2017 Guo et al.

  7. Tuning Chocolate Flavor through Development of Thermotolerant Saccharomyces cerevisiae Starter Cultures with Increased Acetate Ester Production

    Science.gov (United States)

    Meersman, Esther; Steensels, Jan; Struyf, Nore; Paulus, Tinneke; Saels, Veerle; Mathawan, Melissa; Allegaert, Leen; Vrancken, Gino

    2015-01-01

    Microbial starter cultures have extensively been used to enhance the consistency and efficiency of industrial fermentations. Despite the advantages of such controlled fermentations, the fermentation involved in the production of chocolate is still a spontaneous process that relies on the natural microbiota at cocoa farms. However, recent studies indicate that certain thermotolerant Saccharomyces cerevisiae cultures can be used as starter cultures for cocoa pulp fermentation. In this study, we investigate the potential of specifically developed starter cultures to modulate chocolate aroma. Specifically, we developed several new S. cerevisiae hybrids that combine thermotolerance and efficient cocoa pulp fermentation with a high production of volatile flavor-active esters. In addition, we investigated the potential of two strains of two non-Saccharomyces species that produce very large amounts of fruity esters (Pichia kluyveri and Cyberlindnera fabianii) to modulate chocolate aroma. Gas chromatography-mass spectrometry (GC-MS) analysis of the cocoa liquor revealed an increased concentration of various flavor-active esters and a decrease in spoilage-related off-flavors in batches inoculated with S. cerevisiae starter cultures and, to a lesser extent, in batches inoculated with P. kluyveri and Cyb. fabianii. Additionally, GC-MS analysis of chocolate samples revealed that while most short-chain esters evaporated during conching, longer and more-fat-soluble ethyl and acetate esters, such as ethyl octanoate, phenylethyl acetate, ethyl phenylacetate, ethyl decanoate, and ethyl dodecanoate, remained almost unaffected. Sensory analysis by an expert panel confirmed significant differences in the aromas of chocolates produced with different starter cultures. Together, these results show that the selection of different yeast cultures opens novel avenues for modulating chocolate flavor. PMID:26590272

  8. Discovery by organism based high-throughput screening of new multi-stage compounds affecting Schistosoma mansoni viability, egg formation and production.

    Science.gov (United States)

    Guidi, Alessandra; Lalli, Cristiana; Gimmelli, Roberto; Nizi, Emanuela; Andreini, Matteo; Gennari, Nadia; Saccoccia, Fulvio; Harper, Steven; Bresciani, Alberto; Ruberti, Giovina

    2017-10-01

    Schistosomiasis, one of the most prevalent neglected parasitic diseases affecting humans and animals, is caused by the Platyhelminthes of the genus Schistosoma. Schistosomes are the only trematodes to have evolved sexual dimorphism and the constant pairing with a male is essential for the sexual maturation of the female. Pairing is required for the full development of the two major female organs, ovary and vitellarium that are involved in the production of different cell types such as oocytes and vitellocytes, which represent the core elements of the whole egg machinery. Sexually mature females can produce a large number of eggs each day. Due to the importance of egg production for both life cycle and pathogenesis, there is significant interest in the search for new strategies and compounds not only affecting parasite viability but also egg production. Here we use a recently developed high-throughput organism-based approach, based on ATP quantitation in the schistosomula larval stage of Schistosoma mansoni for the screening of a large compound library, and describe a pharmacophore-based drug selection approach and phenotypic analyses to identify novel multi-stage schistosomicidal compounds. Interestingly, worm pairs treated with seven of the eight compounds identified show a phenotype characterized by defects in eggshell assemblage within the ootype and egg formation with degenerated oocytes and vitelline cells engulfment in the uterus and/or oviduct. We describe promising new molecules that not only impair the schistosomula larval stage but also impact juvenile and adult worm viability and egg formation and production in vitro.

  9. Effect of Continuous Fermentation of High-Sugar Fruit Must on the Viability and Morphology of Immobilized Yeast on White Foam Glass

    Directory of Open Access Journals (Sweden)

    Joanna Skwira

    2008-01-01

    Full Text Available We studied the effect of continuous fermentation of high-sugar fruit must (containing about 320 g/L of total sugars on the viability and morphology of yeast cells. The process was carried out for 2.5 months in a 4-column bioreactor at 22 °C, using the Saccharomyces bayanus S.o./1AD wine yeast strain, which was immobilized on cubes of white foam glass. During the time of continuous fermentation, the apple wine contained 11.4–16.8 % (by volume of alcohol and a total sugar concentration of 49.2–115.4 g/L. Yeast cells isolated from the carrier at the end of continuous fermentation were bigger than the cells before immobilization and were characterized by various shapes, e.g. they were elongated, large and round or pear-shaped. Some cells were connected to other cells in the form of aggregates. Some yeast cells from the second, third and fourth columns showed a substantial number of wrinkles or folds. Moreover, it was observed that yeast from the carrier in the first column was characterized by the highest viability, 70 %. In the fourth column, the percentage of viable cells was only 11 %.

  10. Thermotolerance-induced goblet cell activity confers protection in post-operative gut barrier dysfunction.

    LENUS (Irish Health Repository)

    Ali, Rohana

    2009-06-01

    There is evidence that some level of protection against the adverse sequelae of surgery is provided by induction of thermotolerance; this protective effect was explored by study of several indicators of bowel wall damage in animals exposed to surgical insults. It has been argued that the mechanism of the protective effect of thermotolerance involves heat shock proteins (HSPs). We hypothesized that the protective effect of thermotolerance may be due in part to changes in the bowel wall itself, and we investigated this hypothesis in an experimental rat model.

  11. Respostas Fisiológicas Associadas à Termotolerância em Pintos de Corte de Duas Linhagens por Exposição a Altas Temperaturas Physiologic Response Associated to Thermotolerance in Broiler Chickens of Two Strains for Exposure to High Temperature

    Directory of Open Access Journals (Sweden)

    Nelson José Laurino Dionello

    2002-02-01

    Full Text Available Com o objetivo de se avaliar o comportamento fisiológico das aves, bem como, se ocorre a expressão da proteína do choque térmico (Hsp 70 no fígado e cérebro, como conseqüência da exposição ao estresse por calor, em quatro dias sucessivos (2 a 5 dias de vida, 20 pintos de corte de duas linhagens Pescoço pelado (Na/na e Hubbard-Petersen, foram ou não estressados à temperatura de 36-37°C, durante cinco horas. Foram colhidas amostras de fígado e cérebro, ao final do período de estresse, e analisadas por Western Blotting, para determinação dos níveis de Hsp 70. Os resultados não mostraram associação entre perda de peso corporal e as demais características. Aumentos da variação de temperatura cloacal apresentaram aumentos nos níveis de Hsp70 no tecido hepático e cerebral. Concluiu-se que a concentração de Hsp70 no cérebro, apresentou-se correlacionada com a variação de temperatura cloacal, quando se usou o condicionamento ao calor em pintos de corte, embora não se possa afirmar que seria indicativo de termotolerância de aves.This study was run to evaluate the physiological response and expression of hepatic and brain Hsp70, of 20 Naked neck (Na/na and Hubbard-Petersen broiler chicks (2 to 5 days of age, exposed or not to 36-37°C. The trial was conducted during four days and the birds were exposed to high temperature during 5 hours. Liver and brain tissue samples were collected at the end of the stress period and analyzed for Western Blotting for determination of Hsp70 level. No association between body weight loss and the other variables were observed. Increasing the variation in cloacal temperature resulted in an increase in Hsp70 hepatic and brain tissue levels. Results indicate that brain Hsp70 levels were correlated with variation in cloacal temperature, concerning the chicks subjected to heat stress, but it can not be stated that it is an indicative of thermotolerance by the birds.

  12. Genetic diversity of thermotolerant Campylobacter spp. isolates from different stages of the poultry meat supply chain in Argentina.

    Science.gov (United States)

    Zbrun, María V; Romero-Scharpen, Analía; Olivero, Carolina; Zimmermann, Jorge A; Rossler, Eugenia; Soto, Lorena P; Astesana, Diego M; Blajman, Jesica E; Berisvil, Ayelén; Frizzo, Laureano S; Signorini, Marcelo L

    The objective of this study was to investigate a clonal relationship among thermotolerant Campylobacter spp. isolates from different stages of the poultry meat supply chain in Argentina. A total of 128 thermotolerant Campylobacter spp. (89 C. jejuni and 39 C. coli) isolates from six poultry meat chains were examined. These isolates were from: a) hens from breeder flocks, b) chickens on the farm (at ages 1 wk and 5 wk), c) chicken carcasses in the slaughterhouse, and d) chicken carcasses in the retail market. Chickens sampled along each food chain were from the same batch. Campylobacter spp. isolates were analyzed using pulsed-field gel electrophoresis to compare different profiles according to the source. Clustering of C. jejuni isolates resulted in 17 profiles, with four predominant genotypes and many small profiles with just a few isolates or unique patterns, showing a very high degree of heterogeneity among the C. jejuni isolates. Some clusters included isolates from different stages within the same chain, which would indicate a spread of strains along the same poultry meat chain. Moreover, twenty-two strains of C. coli clustered in seven groups and the remaining 17 isolates exhibited unique profiles. Evidence for transmission of thermotolerant Campylobacter spp. through the food chain and cross contamination in the slaughterhouses were obtained. This collective evidence should be considered as the scientific basis to implement risk management measures to protect the public health. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  13. Screening of thermotolerant microorganisms and application for oil separation from palm oil mill wastewater

    National Research Council Canada - National Science Library

    Aran H-Kittikun; Treetippa Laohaprapanon; Poonsuk Prasertsan

    2007-01-01

    ... 1.19 g and oil and grease 1.07 g. In order to release oil and grease trapped in palm fiber debris in the POMW, cellulase- and/or xylanase-enzyme-producing and thermotolerant microorganisms wereisolated...

  14. Bacterial cultures capable of facultative growth on methane under thermophilic or thermotolerant conditions

    National Research Council Canada - National Science Library

    Laursen, Andrew E; Kulpa, Charles F; Niedzielski, Maksymilian F; Estable, Mario C

    2007-01-01

    ...) are capable of growth on methane as a sole carbon source under thermotolerant (OS and 44) to thermophilic (SA) conditions. All three grew better on enriched medium, suggesting methane is not a preferred substrate...

  15. Acquired Thermotolerance and Heat Shock Proteins in Thermophiles from the Three Phylogenetic Domains

    DEFF Research Database (Denmark)

    Trent, Jonathan D.; Gabrielsen, Mette; Jensen, Bo

    1994-01-01

    Thermophilic organisms from each of the three phylogenetic domains (Bacteria, Archaea, and Eucarya) acquired thermotolerance after heat shock. Bacillus caldolyticus grown at 60 degrees C and heat shocked at 69 degrees C for 10 min showed thermotolerance at 74 degrees C, Sulfolobus shibatae grown...... at 70 degrees C and heat shocked at 88 degrees C for 60 min showed thermotolerance at 95 degrees C, and Thermomyces lanuginosus grown at 50 degrees C and heat shocked at 55 degrees C for 60 min showed thermotolerance at 58 degrees C. Determinations of protein synthesis during heat shock revealed...... differences in the dominant heat shock proteins for each species. For B. caldolyticus, a 70-kDa protein dominated while for S. shibatae, a 55-kDa protein dominated and for T. lanuginosus, 31- to 33-kDa proteins dominated. Reagents that disrupted normal protein synthesis during heat shock prevented...

  16. Structural systems biology evaluation of metabolic thermotolerance in Escherichia coli.

    Science.gov (United States)

    Chang, Roger L; Andrews, Kathleen; Kim, Donghyuk; Li, Zhanwen; Godzik, Adam; Palsson, Bernhard O

    2013-06-07

    Genome-scale network reconstruction has enabled predictive modeling of metabolism for many systems. Traditionally, protein structural information has not been represented in such reconstructions. Expansion of a genome-scale model of Escherichia coli metabolism by including experimental and predicted protein structures enabled the analysis of protein thermostability in a network context. This analysis allowed the prediction of protein activities that limit network function at superoptimal temperatures and mechanistic interpretations of mutations found in strains adapted to heat. Predicted growth-limiting factors for thermotolerance were validated through nutrient supplementation experiments and defined metabolic sensitivities to heat stress, providing evidence that metabolic enzyme thermostability is rate-limiting at superoptimal temperatures. Inclusion of structural information expanded the content and predictive capability of genome-scale metabolic networks that enable structural systems biology of metabolism.

  17. Laccase produced by a thermotolerant strain of Trametes trogii LK13

    OpenAIRE

    Jinping Yan; Yuhui Chen; Jiezhen Niu; Daidi Chen; Irbis Chagan

    2015-01-01

    Thermophilic and thermotolerant micro-organisms strains have served as the natural source of industrially relevant and thermostable enzymes. Although some strains of the Trametes genus are thermotolerant, few Trametes strains were studied at the temperature above 30 °C until now. In this paper, the laccase activity and the mycelial growth rate for Trametes trogii LK13 are superior at 37 °C. Thermostability and organic cosolvent tolerance assays of the laccase produced at 37 °C indicated that ...

  18. Production of savinase and population viability of Bacillus clausii during high-cell-density fed-batch cultivations

    DEFF Research Database (Denmark)

    Christiansen, Torben; Michaelsen, S.; Wumpelmann, M.

    2003-01-01

    The growth and product formation of a Savinase-producing Bacillus clausii were investigated in high-cell-density fed-batch cultivations with both linear and exponential feed profiles. The highest specific productivity of Savinase was observed shortly after the end of the initial batch phase for all....... The physiological state of the cells was monitored during the cultivations using a flow cytometry assay based on the permeability of the cell membrane to propidium iodide. In the latter parts of the fed-batch cultures with a linear feed profile, a large portion of the cell population was found to have a permeable...

  19. [Cultural detection of thermotolerant Campylobacter spp. in food--potentials and limitations of diagnostic tools in the context of official food control].

    Science.gov (United States)

    Messelhäusser, Ute; Thärigen, Diana; Fella, Christiane; Schreiner, Hermann; Busch, Ulrich; Höller, Christiane

    2015-01-01

    Thermotolerant Campylobacter spp. rank among the most important foodborne pathogens in Germany. Therefore a necessity for rapid and routinely useable detection methods exists also in the area of food microbiology. A reliable, cultura qualitative, but also quantitative detection of thermotolerant Campylobacter spp. pose a challenge, at least concerning special food matrices, especially because in the context of official food control the cultural detection of thermotolerant Campylobacter spp. is needed. This was the reason, why different cultural detection methods, beside the standard procedure of ISO 10272:2006, in combination with molecular and immunological screening methods were tested at the Bavarian Health and Food Safety Authority (LGL) during the last years for the use in routine diagnostic using different food matrices of animal and plant origin. The results of the comparative studies showed clearly that no enrichment broth tested gave completely satisfactory results for an only culture-based detection the combination with a screening method is therefore recommended for a rapid and reliable detection. But in this case the user should take into account that the sensitivity of such molecular and immunological methods is normally so high that in some cases, depending on the food matrix and processing step, the isolation of the pathogen would not be possible in samples, which were positive in the screening methods.

  20. Thermotolerance and regeneration in the brittle star species complex Ophioderma longicauda: a preliminary study comparing lineages and Mediterranean basins.

    Science.gov (United States)

    Weber, Alexandra Anh-Thu; Dupont, Sam; Chenuil, Anne

    2013-01-01

    Global warming is expected to change marine species distributions; it is thus critical to understand species current thermotolerance. The brittle star species complex Ophioderma longicauda comprises a broadcast spawning lineage L1 and a brooding lineage L3. We collected L1 specimens from Marseilles and Crete, and L3 specimens from Crete. We monitored survival, autotomy and arm regeneration at 17, 26 and 30°C during 14 weeks. Globally O. longicauda showed good resistance to elevated temperatures compared to other published studies on ophiuroids. The L3 sample displayed a better thermotolerance than L1 samples. Yet, more research is needed to establish whether these differences are due to lineages, geographic origin, or random effects. We provided for the first time individual regeneration trajectories, and showed that regeneration followed a growth curve and was highly influenced by temperature in both lineages. Our results highlight the importance of taking into account the presence of cryptic species when studying the potential effects of global warming. © 2013 Académie des sciences. Published by Elsevier SAS. All rights reserved.

  1. Thermotolerant Kluyveromyces marxianus and Saccharomyces cerevisiae strains representing potentials for bioethanol production from Jerusalem artichoke by consolidated bioprocessing

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Nan [Agricultural Univ., Qingdao, SD (China). College of Animal Science and Technology; Chinese Academy of Sciences, Qingdao, SD (China). Key Lab. of Biofuels; Yuan, Bo; Wang, Shi-An; Li, Fu-Li [Chinese Academy of Sciences, Qingdao, SD (China). Key Lab. of Biofuels; Sun, Juan [Agricultural Univ., Qingdao, SD (China). College of Animal Science and Technology

    2012-09-15

    Thermotolerant inulin-utilizing yeast strains are desirable for ethanol production from Jerusalem artichoke tubers by consolidated bioprocessing (CBP). To obtain such strains, 21 naturally occurring yeast strains isolated by using an enrichment method and 65 previously isolated Saccharomyces cerevisiae strains were investigated in inulin utilization, extracellular inulinase activity, and ethanol fermentation from inulin and Jerusalem artichoke tuber flour at 40 C. The strains Kluyveromyces marxianus PT-1 (CGMCC AS2.4515) and S. cerevisiae JZ1C (CGMCC AS2.3878) presented the highest extracellular inulinase activity and ethanol yield in this study. The highest ethanol concentration in Jerusalem artichoke tuber flour fermentation (200 g L{sup -1}) at 40 C achieved by K. marxianus PT-1 and S. cerevisiae JZ1C was 73.6 and 65.2 g L{sup -1}, which corresponded to the theoretical ethanol yield of 90.0 and 79.7 %, respectively. In the range of 30 to 40 C, temperature did not have a significant effect on ethanol production for both strains. This study displayed the distinctive superiority of K. marxianus PT-1 and S. cerevisiae JZ1C in the thermotolerance and utilization of inulin-type oligosaccharides reserved in Jerusalem artichoke tubers. It is proposed that both K. marxianus and S. cerevisiae have considerable potential in ethanol production from Jerusalem artichoke tubers by a high temperature CBP. (orig.)

  2. Thermotolerant Kluyveromyces marxianus and Saccharomyces cerevisiae strains representing potentials for bioethanol production from Jerusalem artichoke by consolidated bioprocessing.

    Science.gov (United States)

    Hu, Nan; Yuan, Bo; Sun, Juan; Wang, Shi-An; Li, Fu-Li

    2012-09-01

    Thermotolerant inulin-utilizing yeast strains are desirable for ethanol production from Jerusalem artichoke tubers by consolidated bioprocessing (CBP). To obtain such strains, 21 naturally occurring yeast strains isolated by using an enrichment method and 65 previously isolated Saccharomyces cerevisiae strains were investigated in inulin utilization, extracellular inulinase activity, and ethanol fermentation from inulin and Jerusalem artichoke tuber flour at 40 °C. The strains Kluyveromyces marxianus PT-1 (CGMCC AS2.4515) and S. cerevisiae JZ1C (CGMCC AS2.3878) presented the highest extracellular inulinase activity and ethanol yield in this study. The highest ethanol concentration in Jerusalem artichoke tuber flour fermentation (200 g L(-1)) at 40 °C achieved by K. marxianus PT-1 and S. cerevisiae JZ1C was 73.6 and 65.2 g L(-1), which corresponded to the theoretical ethanol yield of 90.0 and 79.7 %, respectively. In the range of 30 to 40 °C, temperature did not have a significant effect on ethanol production for both strains. This study displayed the distinctive superiority of K. marxianus PT-1 and S. cerevisiae JZ1C in the thermotolerance and utilization of inulin-type oligosaccharides reserved in Jerusalem artichoke tubers. It is proposed that both K. marxianus and S. cerevisiae have considerable potential in ethanol production from Jerusalem artichoke tubers by a high temperature CBP.

  3. Economic viability of ultra high-performance fiber reinforced concrete in prestressed concrete wind towers to support a 5 MW turbine

    Directory of Open Access Journals (Sweden)

    P. V. C. N. GAMA

    Full Text Available Abstract The Ultra-High Performance Fiber-Reinforced Concrete is a material with remarkable mechanical properties and durability when compared to conventional and high performance concrete, which allows its use even without the reinforcement. This paper proposes the design of prestressed towers for a 5 MW turbine, through regulatory provisions and the limit states method, with UHPFRC and the concrete class C50, comparing the differences obtained in the design by parametric analysis, giving the advantages and disadvantages of using this new type of concrete. Important considerations, simplifications and notes are made to the calculation process, as well as in obtaining the prestressing and passive longitudinal and passive transverse reinforcement, highlighting the shear strength of annular sections comparing a model proposed here with recent experimental results present in the literature, which was obtained good agreement. In the end, it is estimated a first value within the constraints here made to ensure the economic viability of the use of UHPFRC in a 100 m prestressed wind tower with a 5 MW turbine.

  4. Thermotolerant Yeasts for Bioethanol Production Using Lignocellulosic Substrates

    Science.gov (United States)

    Pasha, Chand; Rao, L. Venkateswar

    glucose without a physical and chemical pre-treatment. The pre-treatment processes normally applied on the different substrates are acidic hydrolysis, steam explosion and wet oxidation. A problem for most pretreatment methods is the generation of compounds that are inhibitory towards the fermenting microorganisms, primarily phenols. Degradation products that could have inhibitory action in later fermentation steps are avoided during pre-treatment by wet oxidation. Followed by pre treatment, hydrolysed with enzymes known as cellulases and hemicellulases, which hydrolyse cellulose and hemicellulose respectively. The production of bioethanol requires two steps, fermentation and distillation. Practically all ethanol fermentation is still based on Saccharomyces cerevisiae . The fermentation using thermotolerant yeasts has more advantageous in that they have faster fermentation rates, avoid the cooling costs, and decrease the over all fermentation costs, so that ethanol can be made available at cheaper rates. In addition they can be used for efficient simultaneous saccharification and fermentation of cellulose by cellulases because the temperature optimum of cellulase enzymes (about 40 ° C to 45 ° C) is close to the fermentation temperature of thermotolerant yeasts. Hence selection and improvement of thermotolerant yeasts for bioconversion of lignocellulosic substrates is very useful.

  5. Viability, invariance and applications

    CERN Document Server

    Carja, Ovidiu; Vrabie, Ioan I

    2007-01-01

    The book is an almost self-contained presentation of the most important concepts and results in viability and invariance. The viability of a set K with respect to a given function (or multi-function) F, defined on it, describes the property that, for each initial data in K, the differential equation (or inclusion) driven by that function or multi-function) to have at least one solution. The invariance of a set K with respect to a function (or multi-function) F, defined on a larger set D, is that property which says that each solution of the differential equation (or inclusion) driven by F and issuing in K remains in K, at least for a short time.The book includes the most important necessary and sufficient conditions for viability starting with Nagumo's Viability Theorem for ordinary differential equations with continuous right-hand sides and continuing with the corresponding extensions either to differential inclusions or to semilinear or even fully nonlinear evolution equations, systems and inclusions. In th...

  6. Pigment production by a new thermotolerant microalga Coelastrella sp. F50.

    Science.gov (United States)

    Hu, Che-Wei; Chuang, Lu-Te; Yu, Po-Chien; Chen, Ching-Nen Nathan

    2013-06-15

    Microalgae are good crops to produce natural pigments because of their high growth rates. Tropical zones are better locations than temperate areas for microalgal cultivation because they have longer duration of daylight and more stable temperatures throughout the year, but the high temperatures pose a challenge to microalgal cultivation. A newly isolated thermotolerant microalga produces reddish pigments under environmental stress. Morphological and molecular evidence including meridional ribs on the cell wall, pigment production, and its 18S rDNA sequence suggests that this microalga belongs to the genus Coelastrella. Salt stress and high light intensity accelerated biosynthesis of the pigments, and significant quantities of oil accumulated as the cells experienced stress due to nutrient deficiency. This microalga could withstand temperature of 50°C for more than 8h, which is a necessary trait for outdoor cultivation in tropical areas. The pigments contain astaxanthin, lutein, canthaxanthin, and β-carotene as analysed by using HPLC. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. Effect of magnetic nanoparticle heating on cortical neuron viability.

    Science.gov (United States)

    Rivet, Christopher J; Yuan, Yuan; Gilbert, Ryan J; Borca-Tasciuc, Diana-Andra

    2014-03-01

    Superparamagnetic iron oxide nanoparticles are currently approved for use as an adjunctive treatment to glioblastoma multiforme radiotherapy. Radio frequency stimulation of the nanoparticles generates localised hyperthermia, which sensitises the tumour to the effects of radiotherapy. Clinical trials reported thus far are promising, with an increase in patient survival rate; however, what are left unaddressed are the implications of this technology on the surrounding healthy tissue. Aminosilane-coated iron oxide nanoparticles suspended in culture medium were applied to chick embryonic cortical neuron cultures. Cultures were heated to 37 °C or 45 °C by an induction coil system for 2 h. The latter regime emulates the therapeutic conditions of the adjunctive therapy. Cellular viability and neurite retraction was quantified 24 h after exposure to the hyperthermic events. The hyperthermic load inflicted little damage to the neuron cultures, as determined by calcein-AM, propidium iodide, and alamarBlue® assays. Fluorescence imaging was used to assess the extent of neurite retraction which was found to be negligible. Retention of chick, embryonic cortical neuron viability was confirmed under the thermal conditions produced by radiofrequency stimulation of iron oxide nanoparticles. While these results are not directly applicable to clinical applications of hyperthermia, the thermotolerance of chick embryonic cortical neurons is promising and calls for further studies employing human cultures of neurons and glial cells.

  8. Oxygen availability strongly affects chronological lifespan and thermotolerance in batch cultures of Saccharomyces cerevisiae

    Science.gov (United States)

    Bisschops, Markus M.; Vos, Tim; Martínez-Moreno, Rubén; Cortés, Pilar T.; Pronk, Jack T.; Daran-Lapujade, Pascale

    2015-01-01

    Stationary-phase (SP) batch cultures of Saccharomyces cerevisiae, in which growth has been arrested by carbon-source depletion, are widely applied to study chronological lifespan, quiescence and SP-associated robustness. Based on this type of experiments, typically performed under aerobic conditions, several roles of oxygen in aging have been proposed. However, SP in anaerobic yeast cultures has not been investigated in detail. Here, we use the unique capability of S. cerevisiae to grow in the complete absence of oxygen to directly compare SP in aerobic and anaerobic bioreactor cultures. This comparison revealed strong positive effects of oxygen availability on adenylate energy charge, longevity and thermotolerance during SP. A low thermotolerance of anaerobic batch cultures was already evident during the exponential growth phase and, in contrast to the situation in aerobic cultures, was not substantially increased during transition into SP. A combination of physiological and transcriptome analysis showed that the slow post-diauxic growth phase on ethanol, which precedes SP in aerobic, but not in anaerobic cultures, endowed cells with the time and resources needed for inducing longevity and thermotolerance. When combined with literature data on acquisition of longevity and thermotolerance in retentostat cultures, the present study indicates that the fast transition from glucose excess to SP in anaerobic cultures precludes acquisition of longevity and thermotolerance. Moreover, this study demonstrates the importance of a preceding, calorie-restricted conditioning phase in the acquisition of longevity and stress tolerance in SP yeast cultures, irrespective of oxygen availability. PMID:28357268

  9. Evolution of thermotolerance in hot spring cyanobacteria of the genus Synechococcus

    Science.gov (United States)

    Miller, S. R.; Castenholz, R. W.

    2000-01-01

    The extension of ecological tolerance limits may be an important mechanism by which microorganisms adapt to novel environments, but it may come at the evolutionary cost of reduced performance under ancestral conditions. We combined a comparative physiological approach with phylogenetic analyses to study the evolution of thermotolerance in hot spring cyanobacteria of the genus Synechococcus. Among the 20 laboratory clones of Synechococcus isolated from collections made along an Oregon hot spring thermal gradient, four different 16S rRNA gene sequences were identified. Phylogenies constructed by using the sequence data indicated that the clones were polyphyletic but that three of the four sequence groups formed a clade. Differences in thermotolerance were observed for clones with different 16S rRNA gene sequences, and comparison of these physiological differences within a phylogenetic framework provided evidence that more thermotolerant lineages of Synechococcus evolved from less thermotolerant ancestors. The extension of the thermal limit in these bacteria was correlated with a reduction in the breadth of the temperature range for growth, which provides evidence that enhanced thermotolerance has come at the evolutionary cost of increased thermal specialization. This study illustrates the utility of using phylogenetic comparative methods to investigate how evolutionary processes have shaped historical patterns of ecological diversification in microorganisms.

  10. Oxygen availability strongly affects chronological lifespan and thermotolerance in batch cultures of Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Markus M.M. Bisschops

    2015-10-01

    Full Text Available Stationary-phase (SP batch cultures of Saccharomyces cerevisiae, in which growth has been arrested by carbon-source depletion, are widely applied to study chronological lifespan, quiescence and SP-associated robustness. Based on this type of experiments, typically performed under aerobic conditions, several roles of oxygen in aging have been proposed. However, SP in anaerobic yeast cultures has not been investigated in detail. Here, we use the unique capability of S. cerevisiae to grow in the complete absence of oxygen to directly compare SP in aerobic and anaerobic bioreactor cultures. This comparison revealed strong positive effects of oxygen availability on adenylate energy charge, longevity and thermotolerance during SP. A low thermotolerance of anaerobic batch cultures was already evident during the exponential growth phase and, in contrast to the situation in aerobic cultures, was not substantially increased during transition into SP. A combination of physiological and transcriptome analysis showed that the slow post-diauxic growth phase on ethanol, which precedes SP in aerobic, but not in anaerobic cultures, endowed cells with the time and resources needed for inducing longevity and thermotolerance. When combined with literature data on acquisition of longevity and thermotolerance in retentostat cultures, the present study indicates that the fast transition from glucose excess to SP in anaerobic cultures precludes acquisition of longevity and thermotolerance. Moreover, this study demonstrates the importance of a preceding, calorie-restricted conditioning phase in the acquisition of longevity and stress tolerance in SP yeast cultures, irrespective of oxygen availability.

  11. Effect of heat shock protein 70 polymorphism on thermotolerance in Tharparkar cattle

    Directory of Open Access Journals (Sweden)

    Sandip Bhat

    2016-02-01

    Full Text Available Aim: Out of various members of heat shock protein (HSP superfamily which act a molecular chaperon by binding to the denaturing protein thus stabilizing them and preserving their activity, HSP70 are of major importance in thermotolerance development. Thus, present investigation aimed at a screening of HSP70 gene for polymorphisms and possible differences in thermotolerance in Tharparkar breed of cattle. Materials and Methods: A 295 bp fragment of HSP70 gene was subjected to polymerase chain reaction-singlestrand conformation polymorphism (SSCP followed by sequencing of different SSCP patterns in 64 Tharparkar cattle. A comparative thermotolerance of identified genotypes was analyzed using heat tolerance coefficients (HTCs of animals for different seasons. Results: Three SSCP patterns and consequently two alleles namely A and B were documented in one fragment of HSP70 gene. On sequencing, one single-nucleotide polymorphism with G > T substitution was found at a position that led to a change of amino acid aspartate to tyrosine in allele A. It was found that in maintaining near normal average rectal temperature, genotype AA was superior (p≤0.01. Genotype AA, thus, was found to be most thermotolerant genotype with the highest HTC (p≤0.01. Conclusion: The polymorphism at HSP70 is expected to be a potent determinant for heat tolerance in cattle, which may aid in selection for thermotolerance in cattle.

  12. Transcriptome analysis of the thermotolerant yeast Kluyveromyces marxianus CCT 7735 under ethanol stress.

    Science.gov (United States)

    Diniz, Raphael Hermano Santos; Villada, Juan C; Alvim, Mariana Caroline Tocantins; Vidigal, Pedro Marcus Pereira; Vieira, Nívea Moreira; Lamas-Maceiras, Mónica; Cerdán, María Esperanza; González-Siso, María-Isabel; Lahtvee, Petri-Jaan; da Silveira, Wendel Batista

    2017-09-01

    The thermotolerant yeast Kluyveromyces marxianus displays a potential to be used for ethanol production from both whey and lignocellulosic biomass at elevated temperatures, which is highly alluring to reduce the cost of the bioprocess. Nevertheless, contrary to Saccharomyces cerevisiae, K. marxianus cannot tolerate high ethanol concentrations. We report the transcriptional profile alterations in K. marxianus under ethanol stress in order to gain insights about mechanisms involved with ethanol response. Time-dependent changes have been characterized under the exposure of 6% ethanol and compared with the unstressed cells prior to the ethanol addition. Our results reveal that the metabolic flow through the central metabolic pathways is impaired under the applied ethanol stress. Consistent with these results, we also observe that genes involved with ribosome biogenesis are downregulated and gene-encoding heat shock proteins are upregulated. Remarkably, the expression of some gene-encoding enzymes related to unsaturated fatty acid and ergosterol biosynthesis decreases upon ethanol exposure, and free fatty acid and ergosterol measurements demonstrate that their content in K. marxianus does not change under this stress. These results are in contrast to the increase previously reported with S. cerevisiae subjected to ethanol stress and suggest that the restructuration of K. marxianus membrane composition differs in the two yeasts which gives important clues to understand the low ethanol tolerance of K. marxianus compared to S. cerevisiae.

  13. POSSIBLE ROLE OF LOCALIZED PROTEIN DENATURATION IN THE MECHANISM OF INDUCTION OF THERMOTOLERANCE BY HEAT, SODIUM-ARSENITE AND ETHANOL

    NARCIS (Netherlands)

    BURGMAN, PWJJ; KAMPINGA, HH; KONINGS, AWT

    1993-01-01

    Heat, sodium-arsenite, and ethanol-induced thermotolerance are compared, especially with regard to the induced resistance of proteins of the particulate fraction (PF) against heat-induced denaturation. While all three agents induce thermotolerance as expressed as an enhanced survival after

  14. Natural variation in the thermotolerance of neural function and behavior due to a cGMP-dependent protein kinase.

    Directory of Open Access Journals (Sweden)

    Ken Dawson-Scully

    Full Text Available Although it is acknowledged that genetic variation contributes to individual differences in thermotolerance, the specific genes and pathways involved and how they are modulated by the environment remain poorly understood. We link natural variation in the thermotolerance of neural function and behavior in Drosophila melanogaster to the foraging gene (for, which encodes a cGMP-dependent protein kinase (PKG as well as to its downstream target, protein phosphatase 2A (PP2A. Genetic and pharmacological manipulations revealed that reduced PKG (or PP2A activity caused increased thermotolerance of synaptic transmission at the larval neuromuscular junction. Like synaptic transmission, feeding movements were preserved at higher temperatures in larvae with lower PKG levels. In a comparative assay, pharmacological manipulations altering thermotolerance in a central circuit of Locusta migratoria demonstrated conservation of this neuroprotective pathway. In this circuit, either the inhibition of PKG or PP2A induced robust thermotolerance of neural function. We suggest that PKG and therefore the polymorphism associated with the allelic variation in for may provide populations with natural variation in heat stress tolerance. for's function in behavior is conserved across most organisms, including ants, bees, nematodes, and mammals. PKG's role in thermotolerance may also apply to these and other species. Natural variation in thermotolerance arising from genes involved in the PKG pathway could impact the evolution of thermotolerance in natural populations.

  15. Acquisition of thermotolerant yeast Saccharomyces cerevisiae by breeding via stepwise adaptation.

    Science.gov (United States)

    Satomura, Atsushi; Katsuyama, Yoshiaki; Miura, Natsuko; Kuroda, Kouichi; Tomio, Ayako; Bamba, Takeshi; Fukusaki, Eiichiro; Ueda, Mitsuyoshi

    2013-01-01

    A thermotolerant Saccharomyces cerevisiae yeast strain, YK60-1, was bred from a parental strain, MT8-1, via stepwise adaptation. YK60-1 grew at 40°C, a temperature at which MT8-1 could not grow at all. YK60-1 exhibited faster growth than MT8-1 at 30°C. To investigate the mechanisms how MT8-1 acquired thermotolerance, DNA microarray analysis was performed. The analysis revealed the induction of stress-responsive genes such as those encoding heat shock proteins and trehalose biosynthetic enzymes in YK60-1. Furthermore, nontargeting metabolome analysis showed that YK60-1 accumulated more trehalose, a metabolite that contributes to stress tolerance in yeast, than MT8-1. In conclusion, S. cerevisiae MT8-1 acquired thermotolerance by induction of specific stress-responsive genes and enhanced intracellular trehalose levels. © 2013 American Institute of Chemical Engineers.

  16. ISOLATION AND IDENTIFICATION OF A THERMOTOLERANT PLANT GROWTH PROMOTING PSEUDOMONAS PUTIDA PRODUCING TREHALOSE SYNTHASE

    Directory of Open Access Journals (Sweden)

    Ali Sk.Z.

    2013-08-01

    Full Text Available A thermotolerant plant growth promoting Pseudomonas isolate growing at 40oC producing trehalose synthase (TreS was isolated from rhizosphere soil under semi arid conditions of India. Trehalose synthase was extracted; purified and enzymatic activity was examined at various temperatures and pH. The optimum temperature and pH was 38oC and pH 7.5 and the activity declined at above or below the optimum pH and temperature. The enzyme was active on maltose and trehalose among saccharides tested. The enzyme had a higher catalytic activity for maltose with a trehalose yield of 72% than for trehalose where 30% yield of maltose was achieved, indicating maltose as preferred substrate. The isolate showed multiple plant growth promoting traits (indole acetic acid (IAA, phosphate solubilization, siderophore and ammonia both at ambient (28oC and high temperature (40oC. Based on phenotypic and 16SrRNA analysis the isolate was identified as Pseudomonas putida (Accession No. GU396283.

  17. Predicting virus evolution: the relationship between genetic robustness and evolvability of thermotolerance.

    Science.gov (United States)

    Ogbunugafor, C B; McBride, R C; Turner, P E

    2009-01-01

    Evolutionary biologists often seek to infer historical patterns of relatedness among organisms using phylogenetic methods and to gauge the evolutionary processes that determine variation among individuals in extant populations. But relatively less effort is devoted to making evolutionary biology a truly predictive science, where future evolutionary events are precisely foreseen. Accurate predictions of evolvability would be particularly useful in the evolution of infectious diseases, such as the ability to preemptively address the challenge of pathogens newly emerging in humans and other host populations. Experimental evolution of microbes allows the possibility to rigorously test hypotheses regarding pathogen evolvability. Here, we review how genetic robustness was a useful predictor in gauging which variants of RNA virus varphi6 should evolve faster in a novel high-temperature environment. We also present new data on the relative survival of robust and brittle viruses across elevated temperatures and durations of ultraviolet exposure, to infer a possible mechanism for robustness. Our work suggests that virus adaptability in a new environment can be predicted given knowledge of virus canalization in the face of mutational input. These results hint that accurate predictions of virus evolvability are a realistic possibility, at least under circumstances of adaptive thermotolerance.

  18. Purification and Characterization of Organic Solvent and Detergent Tolerant Lipase from Thermotolerant Bacillus sp. RN2

    Directory of Open Access Journals (Sweden)

    Tadahiko Kajiwara

    2010-09-01

    Full Text Available The aim of this study was to characterize the organic solvent and detergent tolerant properties of recombinant lipase isolated from thermotolerant Bacillus sp. RN2 (Lip-SBRN2. The isolation of the lipase-coding gene was achieved by the use of inverse and direct PCR. The complete DNA sequencing of the gene revealed that the lip-SBRN2 gene contains 576 nucleotides which corresponded to 192 deduced amino acids. The purified enzyme was homogeneous with the estimated molecular mass of 19 kDa as determined by SDS-PAGE and gel filtration. The Lip-SBRN2 was stable in a pH range of 9–11 and temperature range of 45–60 °C. The enzyme was a non metallo-monomeric protein and was active against pNP-caprylate (C8 and pNP-laurate (C12 and coconut oil. The Lip-SBRN2 exhibited a high level of activity in the presence of 108% benzene, 102.4% diethylether and 112% SDS. It is anticipated that the organic solvent and detergent tolerant enzyme secreted by Bacillus sp. RN2 will be applicable as catalysts for reaction in the presence of organic solvents and detergents.

  19. Ethanol fermentation by the thermotolerant yeast, Kluyveromyces marxianus TISTR5925, of extracted sap from old oil palm trunk

    Directory of Open Access Journals (Sweden)

    Yoshinori Murata

    2015-05-01

    Full Text Available Palm sap extracted from old oil palm trunks was previously found to contain sugar and nutrients (amino acids and vitamins. Some palm saps contain a low content of sugar due to differences in species or in plant physiology. Here we condensed palm sap with a low content of sugar using flat membrane filtration, then fermented the condensed palm sap at high temperature using the thermotolerant, high ethanol-producing yeast, Kluyveromyces marxianus. Ethanol production under non-optimum conditions was evaluated. Furthermore, the energy required to concentrate the palm sap, and the amount of energy that could be generated from the ethanol, was calculated. The condensation of sugar in sap from palm trunk required for economically viable ethanol production was evaluated.

  20. Development of a monoclonal antibody-based colony blot immunoassay for detection of thermotolerant Campylobacter species.

    Science.gov (United States)

    Huang, Hongsheng; Phipps-Todd, Beverley; McMahon, Tanis; Elmgren, Catherine L; Lutze-Wallace, Cheryl; Todd, Zoe A; Garcia, Manuel M

    2016-11-01

    Campylobacter species, particularly thermotolerant Campylobacter spp., such as C. jejuni, are major human foodborne pathogens. Culture methods have been routinely used for the detection of this organism in various types of samples. An alternative, simple and rapid confirmation test(s) without further tedious biochemical tests would be useful. Meanwhile, Campylobacter-like colonies can be difficult to identify on agar plates overgrown with competitive bacteria, which can lead to false-negative results. This study was to develop a simple colony blot immunoassay using a new monoclonal antibody (Mab) produced in the present study for rapid screening, confirmation and quantification of campylobacters on culture agar plates. The procedure developed in this study was able to specifically detect thermotolerant Campylobacter spp., but not other non-thermotolerant Campylobacter and non-Campylobacter reference strains tested. This assay could detect 105 cells in a single dot. This assay showed 100% correlation with the culture method for the blotted membranes from 21 either chicken meat or vegetable samples experimentally inoculated with thermotolerant campylobacters. Among 101 natural samples of chicken meat (n=44), chicken feces (n=20) and vegetables (n=37), this assay also showed positive for 23 chicken meat and 14 fecal samples that were positive for thermotolerant campylobacters by culture method, and identified four additional suspects that were culture negative. Membranes stored at 4°C for at least 4years could also be used for this assay. The assay developed in this study can be used in quantitative study for immediate or archival usage, and for diagnostic test to preliminarily confirm the presence of thermotolerant Campylobacter on agar plates. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

  1. Occurrence of thermotolerant Campylobacter in raw poultry meat, environmental and pigeon stools collected in open-air markets

    Directory of Open Access Journals (Sweden)

    Alberto Bellio

    2014-08-01

    Full Text Available Campylobacteriosis was the most commonly reported zoonosis for confirmed human cases in European Union during 2011. Poultry meat was very often implicated in Campylobacter infections in humans. In Italy commerce of raw poultry meat is common in open-air markets: these areas can be considered at high risk of bacterial contamination due to the high presence birds like pigeons. The aim of this study was to collect data about the contamination by thermotolerant Campylobacter of raw poultry meat commercialised in open-air markets, of work-surfaces in contact with poultry meat and of pigeon stools sampled in the market areas in Turin, Northern Italy. Between September 2011 and December 2012, 86 raw poultry meat samples, 86 environmental swabs and 108 animal samples were collected in 38 open-air markets. Analysis were carried out according to ISO10272-1:2006 standard. C.coli was detected in 2.3% (2/86 of raw poultry meat samples, whereas no swab (0/86 resulted positive. Of pigeon stool 28% (30/107 was positive for C.jejuni (83.3% C.jejuni subsp. jejuni and 16.7% C.jejuni subsp. doylei. C.jejuni subsp. jejuni was isolated from 1 dead pigeon. Our results showed lower rates of contamination than those reported at retail in Europe. Although samples were collected in areas at high risk of contamination, raw poultry meat and work surfaces reported a low level of presence of thermotolerant Campylobacter. The high percentage of C.jejuni isolated from pigeon stools showed the importance of a continuous application of preventive measures by the food business operators and the surveillance activity by the Competent Authority.

  2. Distinct solubility and cytotoxicity regimes of paclitaxel-loaded cationic liposomes at low and high drug content revealed by kinetic phase behavior and cancer cell viability studies.

    Science.gov (United States)

    Steffes, Victoria M; Murali, Meena M; Park, Yoonsang; Fletcher, Bretton J; Ewert, Kai K; Safinya, Cyrus R

    2017-11-01

    Lipid-based particles are used worldwide in clinical trials as carriers of hydrophobic paclitaxel (PTXL) for cancer chemotherapy, albeit with little improvement over the standard-of-care. Improving efficacy requires an understanding of intramembrane interactions between PTXL and lipids to enhance PTXL solubilization and suppress PTXL phase separation into crystals. We studied the solubility of PTXL in cationic liposomes (CLs) composed of positively charged 2,3-dioleyloxypropyltrimethylammonium chloride (DOTAP) and neutral 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC) as a function of PTXL membrane content and its relation to efficacy. Time-dependent kinetic phase diagrams were generated from observations of PTXL crystal formation by differential-interference-contrast microscopy. Furthermore, a new synchrotron small-angle x-ray scattering in situ methodology applied to DOTAP/DOPC/PTXL membranes condensed with DNA enabled us to detect the incorporation and time-dependent depletion of PTXL from membranes by measurements of variations in the membrane interlayer and DNA interaxial spacings. Our results revealed three regimes with distinct time scales for PTXL membrane solubility: hours for >3 mol% PTXL (low), days for ≈ 3 mol% PTXL (moderate), and ≥20 days for Cell viability experiments on human cancer cell lines using CL PTXL nanoparticles (NPs) in the distinct CL PTXL solubility regimes reveal an unexpected dependence of efficacy on PTXL content in NPs. Remarkably, formulations with lower PTXL content and thus higher stability show higher efficacy than those formulated at the membrane solubility limit of ≈3 mol% PTXL (which has been the focus of most previous physicochemical studies and clinical trials of PTXL-loaded CLs). Furthermore, an additional high-efficacy regime is seen on occasion for liposome compositions with PTXL ≥9 mol% applied to cells at short time scales (hours) after formation. At longer time scales (days), CL PTXL NPs with ≥3

  3. Isolation of thermotolerant campylobacters and C. hyointestinalis from rectal swabs of healthy pigs

    Directory of Open Access Journals (Sweden)

    Mrenoski Slavco

    2007-11-01

    Full Text Available Thermotolerant campylobacters are the most common bacterial etiological agents of human infectious gastroenteritis worldwide. The most frequent isolated species among them are Campylobacter jejuni and C. coli, and less frequent C. upsaliensis and C. lari. Also C. hyointestinalis, that not belong to the group of thermotolerant campylobacters, has been indicate as an agent of human infectious gastroenteritis. Natural reservoir of all named campylobacters is the intestinal tract of many mammals and birds, including domestic animals. In these animals, campylobacters are commonly present as commensals and their feces is considered as a prime source for environmental contamination. Unlike the human feces which is usually examined in the cases of diarrhea, thermotolerant campylobacters and C. hyointestinalis in the animal feces are generally present in a much lesser amount and the isolation very often could be unsuccessful. The aim of this study was to estimate the validity of applied procedure for isolation (and identification of thermotolerant campylobacters and C. hyointestinalis from pig rectal swabs, as a procedure for detection of healthy animal carriers.

  4. Heat shock proteins (HSP-72 kd) in thermotolerant rat sciatic nerves

    NARCIS (Netherlands)

    Hoogeveen, J. F.; van der Kracht, A. H.; Wondergem, J.; Haveman, J.

    1993-01-01

    Localized heating of the rat sciatic nerve over a length of 5 mm for 30 min at 43 degrees C resulted in the production of heat shock protein 72 kd in every nucleated cell and in the induction of thermotolerance in the heated area. HSP-72 kd was never detected in axons. Heat treatment (30 min, 45

  5. Thermotolerant yeasts selected by adaptive evolution express heat stress response at 30ºC

    DEFF Research Database (Denmark)

    Caspeta, Luis; Chen, Yun; Nielsen, Jens

    2016-01-01

    Exposure to long-term environmental changes across >100s of generations results in adapted phenotypes, but little is known about how metabolic and transcriptional responses are optimized in these processes. Here, we show that thermotolerant yeast strains selected by adaptive laboratory evolution ...

  6. THERMOTOLERANCE IN MAMMALIAN-CELLS - PROTEIN DENATURATION AND AGGREGATION, AND STRESS PROTEINS

    NARCIS (Netherlands)

    KAMPINGA, HH

    Cells that have been pre-exposed to thermal stress can acquire a transient resistance against the killing effect of a subsequent thermal stress. The cause for this phenomenon, called thermotolerance, seems to be an enhanced resistance of proteins against thermal denaturation and aggregation. This

  7. Bacillus methanolicus sp. nov., a New Species of Thermotolerant, Methanol-Utilizing, Endospore-Forming Bacteria

    NARCIS (Netherlands)

    Arfman, Nico; Dijkhuizen, Lubbert; Kirchhof, Gudrun; Ludwig, Wolfgang; Schleifer, Karl-Heinz; Bulygina, Eugenia S.; Chumakov, Konstantin M.; Govorukhina, Natalya I.; Trotsenko, Yuri A.; White, Duncan; Sharp, Richard J.

    1992-01-01

    The generic position of 14 strains of gram-positive bacteria able to use methanol as a growth substrate was determined. All are obligately aerobic, thermotolerant organisms that are able to grow at temperatures of 35 to 60°C. Nine of the strains produce oval spores at a subterminal-to-central

  8. Decolorization of a recalcitrant organic compound (Melanoidin by a novel thermotolerant yeast, Candida tropicalis RG-9

    Directory of Open Access Journals (Sweden)

    Tiwari Soni

    2012-06-01

    Full Text Available Abstract Background Sugarcane distilleries use molasses for ethanol production and generate large volume of effluent containing high biological oxygen demand (BOD and chemical oxygen demand (COD along with melanoidin pigment. Melanoidin is a recalcitrant compound that causes several toxic effects on living system, therefore, may be treated before disposal. The aim of this study was to isolate a potential thermotolerant melanoidin decolorizing yeast from natural resources, and optimized different physico-chemical and nutritional parameters. Results Total 24 yeasts were isolated from the soil samples of near by distillery site, in which isolate Y-9 showed maximum decolorization and identified as Candida tropicalis by Microbial Type Culture Collection (MTCC Chandigarh, India. The decolorization yield was expressed as the decrease in the absorbance at 475 nm against initial absorbance at the same wavelength. Uninoculated medium served as control. Yeast showed maximum decolorization (75% at 45°C using 0.2%, glucose; 0.2%, peptone; 0.05%, MgSO4; 0.01%, KH2PO4; pH-5.5 within 24 h of incubation under static condition. Decolorizing ability of yeast was also confirmed by high performance liquid chromatography (HPLC analysis. Conclusion The yeast strain efficiently decolorized melanoidin pigment of distillery effluent at higher temperature than the other earlier reported strains of yeast, therefore, this strain could also be used at industrial level for melanoidin decolorization as it tolerated a wide range of temperature and pH with very small amount of carbon and nitrogen sources.

  9. The relationship between sperm viability and DNA fragmentation rates.

    Science.gov (United States)

    Samplaski, Mary K; Dimitromanolakis, Apostolos; Lo, Kirk C; Grober, Ethan D; Mullen, Brendan; Garbens, Alaina; Jarvi, Keith A

    2015-05-14

    In humans, sperm DNA fragmentation rates have been correlated with sperm viability rates. Reduced sperm viability is associated with high sperm DNA fragmentation, while conversely high sperm viability is associated with low rates of sperm DNA fragmentation. Both elevated DNA fragmentation rates and poor viability are correlated with impaired male fertility, with a DNA fragmentation rate of >30% indicating subfertility. We postulated that in some men, the sperm viability assay could predict the sperm DNA fragmentation rates. This in turn could reduce the need for sperm DNA fragmentation assay testing, simplifying the infertility investigation and saving money for infertile couples. All men having semen analyses with both viability and DNA fragmentation testing were identified via a prospectively collected database. Viability was measured by eosin-nigrosin assay. DNA fragmentation was measured using the sperm chromosome structure assay. The relationship between DNA fragmentation and viability was assessed using Pearson's correlation coefficient. From 2008-2013, 3049 semen analyses had both viability and DNA fragmentation testing. A strong inverse relationship was seen between sperm viability and DNA fragmentation rates, with r=-0.83. If viability was ≤50% (n=301) then DNA fragmentation was ≥ 30% for 95% of the samples. If viability was ≥75% (n=1736), then the DNA fragmentation was ≤30% for 95% of the patients. Sperm viability correlates strongly with DNA fragmentation rates. In men with high levels of sperm viability≥75%, or low levels of sperm viability≤ 30%, DFI testing may be not be routinely necessary. Given that DNA fragmentation testing is substantially more expensive than vitality testing, this may represent a valuable cost-saving measure for couples undergoing a fertility evaluation.

  10. Cloning, Expression, and Characterization of Thermotolerant Manganese Superoxide Dismutase from Bacillus sp. MHS47

    Directory of Open Access Journals (Sweden)

    Supatra Areekit

    2011-01-01

    Full Text Available A superoxide dismutase gene from thermotolerant Bacillus sp. MHS47 (MnSOD47 was cloned, sequenced, and expressed. The gene has an open reading frame of 612 bp, corresponding to 203 deduced amino acids, with high homology to the amino acid sequences of B. thuringiensis (accession no. EEN01322, B. anthracis (accession no. NP_846724, B. cereus (accession no. ZP_04187911, B. weihenstephanensis (accession no. YP_001646918, and B. pseudomycoides. The conserved manganese-binding sites (H28, H83, D165, and H169 show that MnSOD47 has the specific characteristics of the manganese superoxide dismutase (MnSOD enzymes. MnSOD47 expressed an enzyme with a molecular weight of approximately 22.65 kDa and a specific activity of 3537.75 U/mg. The enzyme is active in the pH range 7–8.5, with an optimum pH of 7.5, and at temperatures in the range 30–45 °C, with an optimum temperature of 37 °C. Tests of inhibitors and metal ions indicated that the enzyme activity is inhibited by sodium azide, but not by hydrogen peroxide or potassium cyanide. These data should benefit future studies of MnSODs in other microorganisms and the biotechnological production of MnSOD47, and could also be used to develop a biosensor for the detection of antioxidants and free radical activity. In the future, this basic knowledge could be applicable to the detection of cancer risks in humans and therapeutic treatments.

  11. Enhanced ethanol production from sugarcane juice by galactose adaptation of a newly isolated thermotolerant strain of Pichia kudriavzevii

    NARCIS (Netherlands)

    Dhaliwal, S.S.; Oberoi, H.S.; Sandhu, S.K.; Nanda, D.; Kumar, D.; Uppal, S.K.

    2011-01-01

    The thermotolerant yeast strain isolated from sugarcane juice through enrichment technique was identified as a strain of Pichiakudriavzevii (Issatchenkiaorientalis) through molecular characterization. The P. kudriavzevii cells adapted to galactose medium produced about 30% more ethanol from

  12. Studies On Fermentation, Alcohol Production And Viability In ...

    African Journals Online (AJOL)

    The reverse was true in the sugarcane bagasse medium. Yeasts with high viability tended to have high alcohol production ability in the sucrose medium and vice-versa. KEY WORDS: Alcohol production; fermentation; induced mutants; Saccharomyces cerevisiae; viability. Global Journal of Pure and Applied Sciences ...

  13. Viability of Baylisascaris procyonis Eggs

    OpenAIRE

    Shira C Shafir; Sorvillo, Frank J.; Sorvillo, Teresa; Eberhard, Mark L.

    2011-01-01

    Infection with Baylisascaris procyonis roundworms is rare but often fatal and typically affects children. We attempted to determine parameters of viability and methods of inactivating the eggs of these roundworms. Loss of viability resulted when eggs were heated to 62°C or desiccated for 7 months but not when frozen at –15°C for 6 months.

  14. Genetic manipulation of Bacillus methanolicus, a gram-positive, thermotolerant methylotroph.

    OpenAIRE

    Cue, D; Lam, H; Dillingham, R L; Hanson, R S; Flickinger, M C

    1997-01-01

    We report the fist genetic transformation system, shuttle vectors, and integrative vectors for the thermotolerant, methylotrophic bacterium Bacillus methanolicus. By using a polyethylene glycol-mediated transformation procedure, we have successfully transformed B. methanolicus with both integrative and multicopy plasmids. For plasmids with a single BmeTI recognition site, dam methylation of plasmid DNA (in vivo or in vitro) was found to enhance transformation efficiency from 7- to 11-fold. Tw...

  15. Trampling Impacts on Thermotolerant Vegetation of Geothermal Areas in New Zealand

    Science.gov (United States)

    Burns, Bruce R.; Ward, Jonet; Downs, Theresa M.

    2013-12-01

    Geothermal features such as geysers, mud pools, sinter terraces, fumaroles, hot springs, and steaming ground are natural attractions often visited by tourists. Visitation rates for such areas in the Taupo Volcanic Zone of New Zealand are in the order of hundreds of thousands annually. These areas are also habitat for rare and specialized plant and microbial communities that live in the steam-heated soils of unusual chemical composition. We evaluated historical and current trampling impacts of tourists on the thermotolerant vegetation of the Waimangu and Waiotapu geothermal areas near Rotorua, and compared the results to experimental trampling at a third site (Taheke) not used by tourists. Historical tourism has removed vegetation and soil from around key features, and remaining subsoil is compacted into an impervious pavement on which vegetation recolonization is unlikely in the short term. Social tracks made by tourists were present at both tourist sites often leading them onto hotter soils than constructed tracks. Vegetation height and cover were lower on and adjacent to social tracks than further from them. Thermotolerant vegetation showed extremely low resistance to experimental trampling. This confirms and extends previous research that also shows that thallophytes and woody shrubs, life forms that dominate in thermotolerant vegetation, are vulnerable to trampling damage. Preservation of these vulnerable ecosystems must ensure that tourist traffic is confined to existing tracks or boardwalks, and active restoration of impacted sites may be warranted.

  16. Thermotolerant coliforms are not a good surrogate for Campylobacter spp. in environmental water.

    Science.gov (United States)

    St-Pierre, Karen; Lévesque, Simon; Frost, Eric; Carrier, Nathalie; Arbeit, Robert D; Michaud, Sophie

    2009-11-01

    This study aimed to assess the importance of quantitatively detecting Campylobacter spp. in environmental surface water. The prevalence and the quantity of Campylobacter spp., thermotolerant coliforms, and Escherichia coli in 2,471 samples collected weekly, over a 2-year period, from 13 rivers and 12 streams in the Eastern Townships, Québec, Canada, were determined. Overall, 1,071 (43%), 1,481 (60%), and 1,463 (59%) samples were positive for Campylobacter spp., thermotolerant coliforms, and E. coli, respectively. There were weak correlations between the weekly distributions of Campylobacter spp. and thermotolerant coliforms (Spearman's rho coefficient = 0.27; P = 0.008) and between the quantitative levels of the two classes of organisms (Kendall tau-b correlation coefficient = 0.233; P coliforms. These findings suggest that microbial monitoring of raw water by using only fecal indicator organisms is not sufficient for assessing the occurrence or the load of thermophilic Campylobacter spp. Insights into the role of environmental water as sources for sporadic Campylobacter infection will require genus-specific monitoring techniques.

  17. Comparison of thermotolerant coliforms and Escherichia coli densities in freshwater bodies.

    Science.gov (United States)

    Hachich, Elayse M; Di Bari, Marisa; Christ, Ana Paula G; Lamparelli, Cláudia C; Ramos, Solange S; Sato, Maria Inês Z

    2012-04-01

    Fecal bacterial indicator analyses have been widely used for monitoring the water quality. This study was designed to determine the ratio between the density of Escherichia coli and other Thermotolerant Coliforms (TtC) bacteria from freshwater samples collected for a two-year period of monitoring. TtC were enumerated by membrane filtration on mFC agar. E. coli enumeration was done by two methods: TtC colonies identified in mFC were inoculated in EC-MUG or water samples were filtered and inoculated in modified mTEC agar media, and both methods were compared for quantitative recovery of E. coli. The results pointed out a mean percentage of E. coli among other thermotolerant coliforms (E. coli/TtC ratio) of 84.3% in mFC media. Taking these results into account, a mandatory standard of 1000 thermotolerant coliforms would correspond to 800 E. coli and the adoption of these E. coli based standards will represent a major improvement for the monitoring of freshwater quality.

  18. Thermotolerant Coliforms Are Not a Good Surrogate for Campylobacter spp. in Environmental Water ▿

    Science.gov (United States)

    St-Pierre, Karen; Lévesque, Simon; Frost, Eric; Carrier, Nathalie; Arbeit, Robert D.; Michaud, Sophie

    2009-01-01

    This study aimed to assess the importance of quantitatively detecting Campylobacter spp. in environmental surface water. The prevalence and the quantity of Campylobacter spp., thermotolerant coliforms, and Escherichia coli in 2,471 samples collected weekly, over a 2-year period, from 13 rivers and 12 streams in the Eastern Townships, Québec, Canada, were determined. Overall, 1,071 (43%), 1,481 (60%), and 1,463 (59%) samples were positive for Campylobacter spp., thermotolerant coliforms, and E. coli, respectively. There were weak correlations between the weekly distributions of Campylobacter spp. and thermotolerant coliforms (Spearman's ρ coefficient = 0.27; P = 0.008) and between the quantitative levels of the two classes of organisms (Kendall tau-b correlation coefficient = 0.233; P coliforms. These findings suggest that microbial monitoring of raw water by using only fecal indicator organisms is not sufficient for assessing the occurrence or the load of thermophilic Campylobacter spp. Insights into the role of environmental water as sources for sporadic Campylobacter infection will require genus-specific monitoring techniques. PMID:19734335

  19. [Presence and viability of Giardia spp. and Cryptosporidium spp. in drinking water and wastewater in the high basin of Bogotá river].

    Science.gov (United States)

    Alarcón, Marlén Andrea; Beltrán, Milena; Cárdenas, Martha Liliana; Campos, María Claudia

    2005-09-01

    Faecal contamination in wastewater and drinking water is linked to the dissemination of water related diseases. The bacteria, virus and parasites present in drinking water are responsible for substantial morbidity and mortality, especially among infants. Giardia spp. and Cryptosporidium spp. were the organisms selected as parasite contamination indicators. Their presence serves as a useful tool for evaluating water quality and determining sanitary risk. At present, in Colombia, concentration and occurrence of these parasites is unknown and an immediate assessment was considered necessary. Protozoan presence was determined in five sampling stations in the Bogotá river upper basin and in two drinking water plants near the same area. The techniques applied for counting encysted forms consisted of inorganic flocculation for wastewater or filtration for drinking water. Fluorogenic vital dyes tested for viability. The presence of Cryptosporidium spp. was confirmed in two of the sampled stations and at two of the drinking water plants. Giardia spp. was found at two of the drinking water plants but not at the sampled stations. Viable cysts were found for Cryptosporidium spp. in one of the samples from the Bogotá river, but only inviable exemplars were obtained from the drinking water plants. The results revealed protozoan presence in drinking and residual water implying the presence of a potential sanitary hazard.

  20. A rapid, high-throughput viability assay for Blastocystis spp. reveals metronidazole resistance and extensive subtype-dependent variations in drug susceptibilities.

    Science.gov (United States)

    Mirza, Haris; Teo, Joshua D W; Upcroft, Jacqui; Tan, Kevin S W

    2011-02-01

    Blastocystis is an emerging protistan parasite of controversial pathogenesis. Although metronidazole (Mz) is standard therapy for Blastocystis infections, there have been accumulating reports of treatment failure, suggesting the existence of drug-resistant isolates. Furthermore, very little is known about Blastocystis susceptibility to standard antimicrobials. In the present study, we established resazurin and XTT viability microassays for Blastocystis spp. belonging to subtypes 4 and 7, both of which have been suggested to represent pathogenic zoonotic subtypes. The optimized resazurin assay was used to screen a total of 19 compounds against both subtypes. Interestingly, subtype 7 parasites were resistant to Mz, a 1-position-substituted 5-nitroimidazole (5-NI), while subtype 4 parasites were sensitive. Some cross-resistance was observed to tinidazole, another 1-position 5-NI. Conversely, subtype 4 parasites were resistant to emetine, while subtype 7 parasites were sensitive. Position 2 5-NIs were effective against both subtypes, as were ornidazole, nitazoxanide, furazolidone, mefloquine, quinicrine, quinine, cotrimoxazole (trimethoprim-sulfamethoxazole), and iodoacetamide. Both subtypes were resistant to chloroquine, doxycycline, paromomycin, ampicillin, and pyrimethamine. This is the first study to report extensive variations in drug sensitivities among two clinically important subtypes. Our study highlights the need to reevaluate established treatment regimens for Blastocystis infections and offers clear new treatment options for Mz treatment failures.

  1. Viability of mesenchymal stem cells during electrospinning

    Directory of Open Access Journals (Sweden)

    G. Zanatta

    2012-02-01

    Full Text Available Tissue engineering is a technique by which a live tissue can be re-constructed and one of its main goals is to associate cells with biomaterials. Electrospinning is a technique that facilitates the production of nanofibers and is commonly used to develop fibrous scaffolds to be used in tissue engineering. In the present study, a different approach for cell incorporation into fibrous scaffolds was tested. Mesenchymal stem cells were extracted from the wall of the umbilical cord and mononuclear cells from umbilical cord blood. Cells were re-suspended in a 10% polyvinyl alcohol solution and subjected to electrospinning for 30 min under a voltage of 21 kV. Cell viability was assessed before and after the procedure by exclusion of dead cells using trypan blue staining. Fiber diameter was observed by scanning electron microscopy and the presence of cells within the scaffolds was analyzed by confocal laser scanning microscopy. After electrospinning, the viability of mesenchymal stem cells was reduced from 88 to 19.6% and the viability of mononuclear cells from 99 to 8.38%. The loss of viability was possibly due to the high viscosity of the polymer solution, which reduced the access to nutrients associated with electric and mechanical stress during electrospinning. These results suggest that the incorporation of cells during fiber formation by electrospinning is a viable process that needs more investigation in order to find ways to protect cells from damage.

  2. Viability of smallholder dairying in Wedza, Zimbabwe.

    Science.gov (United States)

    Zvinorova, Plaxedis Ivy; Halimani, Tinyiko Edward; Mano, Renneth T; Ngongoni, Nobbert Takarwirwa

    2013-04-01

    Viability differences in smallholder dairy farming are a result of differences in access to markets and services. It is hypothesized that innovations that improve productivity and market linkages also improve returns and viability. The viability of smallholder dairying in Wedza was characterised by interviewing 52 households using semi-structured questionnaires. Information on demographics, production, marketing, livestock numbers, assets and constraints was obtained. Farmers were resource-constrained with differences in access to resources. The highly resourced farmers had higher milk output and numbers of livestock. Almost 40 % of the households were female-headed, and these dominated the poor category. Household sizes ranged from 4 to 13 persons. Milk off-take was low (3.7 ± 0.53 l/cow/day), due to various constraints. Only rich farmers had viable enterprises in purely financial terms. Per litre cost of milk was more than selling price (US$0.96) for most farmers except the relatively rich. Operating ratios were 1.7, 0.6, 1.4 and 1.1 for the poor, rich, sub-centre and milk collection centre farmers, respectively. This means incomes from the dairy activities did not cover costs. Sensitivity analysis indicated that increases in total variable costs and labour reduced returns. Milk production and viability were influenced by access to resources and markets.

  3. Physiological ecology of desert biocrust moss following 10 years exposure to elevated CO2: evidence for enhanced photosynthetic thermotolerance

    Science.gov (United States)

    Coe, Kirsten K.; Belnap, Jayne; Grote, Edmund E.; Sparks, Jed P.

    2012-01-01

    In arid regions, biomes particularly responsive to climate change, mosses play an important biogeochemical role as key components of biocrusts. Using the biocrust moss Syntrichia caninervis collected from the Nevada Desert Free Air CO2 Enrichment Facility, we examined the physiological effects of 10 years of exposure to elevated CO2, and the effect of high temperature events on the photosynthetic performance of moss grown in CO2-enriched air. Moss exposed to elevated CO2 exhibited a 46% decrease in chlorophyll, a 20% increase in carbon and no difference in either nitrogen content or photosynthetic performance. However, when subjected to high temperatures (35–40°C), mosses from the elevated CO2 environment showed higher photosynthetic performance and photosystem II (PSII) efficiency compared to those grown in ambient conditions, potentially reflective of a shift in nitrogen allocation to components that offer a higher resistance of PSII to heat stress. This result suggests that mosses may respond to climate change in markedly different ways than vascular plants, and observed CO2-induced photosynthetic thermotolerance in S. caninervis will likely have consequences for future desert biogeochemistry.

  4. Physiological ecology of desert biocrust moss following 10 years exposure to elevated CO₂: evidence for enhanced photosynthetic thermotolerance.

    Science.gov (United States)

    Coe, Kirsten K; Belnap, Jayne; Grote, Edmund E; Sparks, Jed P

    2012-04-01

    In arid regions, biomes particularly responsive to climate change, mosses play an important biogeochemical role as key components of biocrusts. Using the biocrust moss Syntrichia caninervis collected from the Nevada Desert Free Air CO₂ Enrichment Facility, we examined the physiological effects of 10 years of exposure to elevated CO₂, and the effect of high temperature events on the photosynthetic performance of moss grown in CO₂-enriched air. Moss exposed to elevated CO₂ exhibited a 46% decrease in chlorophyll, a 20% increase in carbon and no difference in either nitrogen content or photosynthetic performance. However, when subjected to high temperatures (35-40°C), mosses from the elevated CO₂ environment showed higher photosynthetic performance and photosystem II (PSII) efficiency compared to those grown in ambient conditions, potentially reflective of a shift in nitrogen allocation to components that offer a higher resistance of PSII to heat stress. This result suggests that mosses may respond to climate change in markedly different ways than vascular plants, and observed CO₂-induced photosynthetic thermotolerance in S. caninervis will likely have consequences for future desert biogeochemistry. Copyright © Physiologia Plantarum 2012.

  5. Development of strains of the thermotolerant yeast Hansenula polymorpha capable of alcoholic fermentation of starch and xylan.

    Science.gov (United States)

    Voronovsky, Andriy Y; Rohulya, Olha V; Abbas, Charles A; Sibirny, Andriy A

    2009-01-01

    The thermotolerant yeast Hansenula polymorpha ferments glucose and xylose to ethanol at high temperatures. However, H. polymorpha cannot utilize starchy materials or xylans. Heterologous amylolytic and xylanolytic enzymes have to be expressed in this yeast to provide for utilization and growth on starch and xylan. Genes SWA2 and GAM1 from the yeast Schwanniomyces occidentalis, encoding alpha-amylase and glucoamylase, respectively, were expressed in H. polymorpha. The expression was achieved by integration of the SWA2 and GAM1 genes under the strong constitutive promoter of the H. polymorpha glyceraldehyde-3-phosphate dehydrogenase gene (HpGAP) into H. polymorpha genome. Resulting transformants acquired the ability to grow on a minimal medium containing soluble starch as a sole carbon source. Ethanol production at high-temperature fermentation from starch by the recombinant strains was up to 10 g/L. The XYN2 gene encoding endoxylanase of the fungus Trichoderma reseei was expressed in H. polymorpha. Co-expression of xlnD gene coding for beta-xylosidase of the fungus Aspergillus niger and the XYN2 gene in H. polymorpha was achieved by integration of these genes under control of the HpGAP promoter. Resulting transformants were capable of growth and alcoholic fermentation on a minimal medium supplemented with birchwood xylan as a sole carbon source at 48 degrees C.

  6. Thermotolerance of Leaf Discs from Four Isoprene-Emitting Species Is Not Enhanced by Exposure to Exogenous Isoprene1

    Science.gov (United States)

    Logan, Barry A.; Monson, Russell K.

    1999-01-01

    The effects of exogenously supplied isoprene on chlorophyll fluorescence characteristics were examined in leaf discs of four isoprene-emitting plant species, kudzu (Pueraria lobata [Willd.] Ohwi.), velvet bean (Mucuna sp.), quaking aspen (Populus tremuloides Michx.), and pussy willow (Salix discolor Muhl). Isoprene, supplied to the leaves at either 18 μL L−1 in compressed air or 21 μL L−1 in N2, had no effect on the temperature at which minimal fluorescence exhibited an upward inflection during controlled increases in leaf-disc temperature. During exposure to 1008 μmol photons m−2 s−1 in an N2 atmosphere, 21 μL L−1 isoprene had no effect on the thermally induced inflection of steady-state fluorescence. The maximum quantum efficiency of photosystem II photochemistry decreased sharply as leaf-disc temperature was increased; however, this decrease was unaffected by exposure of leaf discs to 21 μL L−1 isoprene. Therefore, there were no discernible effects of isoprene on the occurrence of symptoms of high-temperature damage to thylakoid membranes. Our data do not support the hypothesis that isoprene enhances leaf thermotolerance. PMID:10398717

  7. Thermotolerance of leaf discs from four isoprene-emitting species is not enhanced by exposure to exogenous isoprene

    Science.gov (United States)

    Logan; Monson

    1999-07-01

    The effects of exogenously supplied isoprene on chlorophyll fluorescence characteristics were examined in leaf discs of four isoprene-emitting plant species, kudzu (Pueraria lobata [Willd.] Ohwi.), velvet bean (Mucuna sp.), quaking aspen (Populus tremuloides Michx.), and pussy willow (Salix discolor Muhl). Isoprene, supplied to the leaves at either 18 &mgr;L L-1 in compressed air or 21 &mgr;L L-1 in N2, had no effect on the temperature at which minimal fluorescence exhibited an upward inflection during controlled increases in leaf-disc temperature. During exposure to 1008 &mgr;mol photons m-2 s-1 in an N2 atmosphere, 21 &mgr;L L-1 isoprene had no effect on the thermally induced inflection of steady-state fluorescence. The maximum quantum efficiency of photosystem II photochemistry decreased sharply as leaf-disc temperature was increased; however, this decrease was unaffected by exposure of leaf discs to 21 &mgr;L L-1 isoprene. Therefore, there were no discernible effects of isoprene on the occurrence of symptoms of high-temperature damage to thylakoid membranes. Our data do not support the hypothesis that isoprene enhances leaf thermotolerance.

  8. Viability of encapsulated Lactobacillus sp. Mar 8

    Directory of Open Access Journals (Sweden)

    EVI TRIANA

    2006-04-01

    Full Text Available Lactobacillus sp. Mar 8 had advantages as probiotic digestive system cholesterol lowering Lactobacillus. Applying in industry, particular processing technique is necessary for gaining product that ready for marketing and consuming. Spray drying is common technique using in various food processing. High processing temperature, 100-200oC, for 3-10 second become the barrier because cells were under extreme temperature stress. Therefore, encapsulate was needed to protect the cells from those extreme conditions. Viability and survival rate of encapsulated Lactobacillus sp. Mar 8 have been investigated. The result showed that Lactobacillus sp. Mar 8 that was encapsulated by 10% skim milk has higher viability than those by 5% skim milk, namely 72.37% and 51.69% respectively. Survival rate of encapsulated Lactobacillus cells will come to zero in 41.28 years. Therefore, encapsulated Lactobacillus sp. Mar 8 may use as probiotic agent.

  9. Bioprospecting thermotolerant ethanologenic yeasts for simultaneous saccharification and fermentation from diverse environments.

    Science.gov (United States)

    Choudhary, Jairam; Singh, Surender; Nain, Lata

    2017-03-01

    Lignocellulosic biomass, a promising renewable energy source, can be used for the production of second generation bioethanol. Simultaneous saccharification and fermentation (SSF), the process which alleviates the problem of separate hydrolysis and fermentation (SHF), requires thermotolerant ethanologenic yeast for bioethanol production. Therefore, ten yeast strains isolated from diverse sources, belonging to various genera like Saccharomyces, Candida, Pichia and Wickerhamomyces were evaluated for their thermotolerance, sugar utilization pattern, inhibitor tolerance and ethanol production potential with glucose, xylose and alkali pretreated paddy straw. All the tested strains were found to be thermotolerant, capable of significant growth at 40°C. Candida tropicalis Y6 was capable of utilizing a wide range of sugars as compared with other yeast isolates. Strains of Candida showed better inhibitor tolerance as compared to Saccharomyces and Pichia strains and exhibited only 5.1-18.8% and 4.7-7.9% reduction in growth with furfural and 5-hydroxymethyl furfural, respectively. Saccharomyces cerevisiae JRC6, isolated from distillery waste, produced ethanol with 88.3% and 89.1% theoretical efficiency at 40°C and 42°C, respectively, from glucose. This strain also produced significantly higher amount of ethanol (3.8 g/L) with better fermentation efficiency (87.9%) from alkali pretreated paddy straw at 40°C, as compared with the other yeast strains. Therefore, S. cerevisiae JRC6, based on its ability to ferment sugars at a higher temperature, can be a promising candidate for production of ethanol from lignocellulosic biomass via SSF process. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  10. Preliminary physiological characteristics of thermotolerant Saccharomyces cerevisiae clinical isolates identified by molecular biology techniques.

    Science.gov (United States)

    Siedlarz, P; Sroka, M; Dyląg, M; Nawrot, U; Gonchar, M; Kus-Liśkiewicz, M

    2016-03-01

    The aim of the study was a molecular identification and physiological characteristic of the five Saccharomyces cerevisiae strains isolated from patients. The tested isolates were compared with control strains (which are of laboratory or commercial origin). The relation of the isolates to baker's yeast S. cerevisiae was studied using species-specific primers in PCR analysis of the ITS-26S region of DNA. Five isolates were genetically identified as the yeast belonging to the genus S. cerevisiae. The effects of temperature and carbon sources on the growth of the yeast strains were analysed. A quantitative characterization of growth kinetics approve that some tested isolates are thermotolerant and are able to grow at range 37-39°C. Among them, one representative is characterized by the highest specific growth rate (0·637 h(-1) ). In conclusions, some strains are defined as potential candidates to use in the biotechnology due to a higher growth rate at elevated temperatures. Screening for further evaluation of biotechnological significance of the tested isolates will be done (e.g. ethanol and trehalose production at higher temperatures). The physiological characterization and confirmation of species identification by molecular methods for yeasts important in the context of biotechnology industry were demonstrated. Thermotolerant microbial strains are required in various industrial applications, for improving productivity and for decreasing the risk of undesirable contaminations when higher temperatures are used. It is important to search for such strains in extreme environments or exotic niches. In this paper, new thermotolerant strains were identified belonging to the Saccharomyces cerevisiae, but differed from typical bakers' yeast, essentially by their growth rate at higher temperature. The described yeast strains are promising for using in biotechnological industry, especially, for production of ethanol and other products at higher temperatures. © 2015 The

  11. Triple Bottom Line Thinking for High Arsenic Bearing Copper-Gold Project in Northern Peru: Assessing the Viability of an Integrated Mine, Mill, and Hydrometallurgical Refinery

    OpenAIRE

    Jonathan Robert Mean

    2011-01-01

    Growth in global urbanization, industrialization and infrastructure expansions will require the development of non-traditional copper resources such as those containing high arsenic to meet the demand for future mine supply. Arsenic levels are rising in current and future mines in resource rich countries such as Peru. Conventional smelting operations cannot accept concentrates high in arsenic (>0.5%) due to process limitations and tightening environmental restrictions. Alternative refining...

  12. Economic viability of ultra high-performance fiber reinforced concrete in prestressed concrete wind towers to support a 5 MW turbine

    OpenAIRE

    GAMA, P. V. C. N.; BITTENCOURT, T. N.

    2017-01-01

    Abstract The Ultra-High Performance Fiber-Reinforced Concrete is a material with remarkable mechanical properties and durability when compared to conventional and high performance concrete, which allows its use even without the reinforcement. This paper proposes the design of prestressed towers for a 5 MW turbine, through regulatory provisions and the limit states method, with UHPFRC and the concrete class C50, comparing the differences obtained in the design by parametric analysis, giving th...

  13. Pleurotus sajor-caju HSP100 complements a thermotolerance ...

    Indian Academy of Sciences (India)

    A putative Hsp100 gene was cloned from the fungus Pleurotus sajor-caju. mRNA expression studies demonstrated that this gene (designated PsHsp100) is highly induced by high temperature, induced less strongly by exposure to ethanol, and not induced by drought or salinity. Heat shock induction is detectable at 37°C ...

  14. Enrichment followed by quantitative PCR both for rapid detection and as a tool for quantitative risk assessment of food-borne thermotolerant campylobacters

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Jacobsen, N. R.; Hoorfar, Jeffrey

    2004-01-01

    As part of a large international project for standardization of PCR (Food-PCR; www.pcr.dk), a multiplex, multiplatform, ready-to-go enrichment followed by a real-time PCR method, including an internal amplification control, was developed for detection of food-borne thermotolerant campylobacters...... Organization (ISO)-based culture method by testing low, medium, and high levels of 12 spiked and 66 unspiked, presumably naturally contaminated, chicken rinse samples. In the RotorGene, a positive PCR response was detected in 40 samples of the 66. This was in complete agreement with the enriched ISO culture...... naturally contaminated chicken samples, which indicates PCR's additional potential as a tool for quantitative risk assessment. Signal from the internal amplification control was detected in all culture-negative samples (VIC Ct: 23.1 to 28.1). The method will be taken further and validated...

  15. Evaluation of high pressure processing, freezing, and fermentation/drying on viability of Trichinella spiralis larvae in raw pork and in Genoa salami

    Science.gov (United States)

    We evaluated high pressure processing (HPP), freezing, and fermentation/drying to inactivate Trichinella spiralis larvae in both infected pig muscle and in Genoa salami produced with trichinae infected pork. In part A, in each of two trials 10 gram portions (2 replicates per treatment) of fresh pig ...

  16. Thermo-tolerant phosphate-solubilizing microbes for multi-functional biofertilizer preparation.

    Science.gov (United States)

    Chang, Cheng-Hsiung; Yang, Shang-Shyng

    2009-02-01

    In order to prepare the multi-functional biofertilizer, thermo-tolerant phosphate-solubilizing microbes including bacteria, actinomycetes, and fungi were isolated from different compost plants and biofertilizers. Except Streptomycesthermophilus J57 which lacked pectinase, all isolates possessed amylase, CMCase, chitinase, pectinase, protease, lipase, and nitrogenase activities. All isolates could solubilize calcium phosphate and Israel rock phosphate; various isolates could solubilize aluminum phosphate, iron phosphate, and hydroxyapatite. During composting, biofertilizers inoculated with the tested microbes had a significantly higher temperature, ash content, pH, total nitrogen, soluble phosphorus content, and germination rate than non-inoculated biofertilizer; total organic carbon and carbon-to-nitrogen ratio showed the opposite pattern. Adding these microbes can shorten the period of maturity, improve the quality, increase the soluble phosphorus content, and enhance the populations of phosphate-solubilizing and proteolytic microbes in biofertilizers. Therefore, inoculating thermo-tolerant phosphate-solubilizing microbes into agricultural and animal wastes represents a practical strategy for preparing multi-functional biofertilizer.

  17. Production of Bioethanol from Carrot Pomace Using the Thermotolerant Yeast Kluyveromyces marxianus

    Energy Technology Data Exchange (ETDEWEB)

    Chi-Yang Yu; Bo-Hong Jiang; Kow-Jen Duan [Tatung University, Tapei, Taiwan (China). Department of Bioengineering

    2013-03-15

    Carrot pomace, a major agricultural waste from the juice industry, was used as a feedstock for bioethanol production by fermentation with the thermotolerant yeast Kluyveromyces marxianus. Treatment of the carrot pomace with Accellerase(TM) 1000 and pectinase at 50 °C for 84 h, resulted in conversion of 42% of its mass to fermentable sugars, mainly glucose, fructose, and sucrose. Simultaneous saccharification and fermentation (SSF) at 42 °C was performed on 10% (w/v) carrot pomace; the concentration of ethanol reached 18 g/L and the yield of ethanol from carrot pomace was 0.18 g/g. The highest ethanol concentration of 37 g/L was observed with an additional charge of 10% supplemented to the original 10% of carrot pomace after 12 h; the corresponding yield was 0.185 g/g. Our results clearly demonstrated the potential of combining a SSF process with thermotolerant yeast for the production of bioethanol using carrot pomace as a feedstock.

  18. Viability of sublethally injured coliform bacteria on fresh-cut cabbage stored in high CO2atmospheres following rinsing with electrolyzed water.

    Science.gov (United States)

    Izumi, Hidemi; Inoue, Ayano

    2018-02-02

    The extent of sublethally injured coliform bacteria on shredded cabbage, either rinsed or not rinsed with electrolyzed water, was evaluated during storage in air and high CO 2 controlled atmospheres (5%, 10%, and 15%) at 5°C and 10°C using the thin agar layer (TAL) method. Sublethally injured coliform bacteria on nonrinsed shredded cabbage were either absent or they were injured at a 64-65% level when present. Rinsing of shredded cabbage with electrolyzed water containing 25ppm available chlorine reduced the coliform counts by 0.4 to 1.1 log and caused sublethal injury ranging from 42 to 77%. Pantoea ananatis was one of the species injured by chlorine stress. When shredded cabbage, nonrinsed or rinsed with electrolyzed water, was stored in air and high CO 2 atmospheres at 5°C for 7days and 10°C for 5days, coliform counts on TAL plates increased from 3.3-4.5 to 6.5-9.0 log CFU/g during storage, with the increase being greater at 10°C than at 5°C. High CO 2 of 10% and 15% reduced the bacterial growth on shredded cabbage during storage at 5°C. Although injured coliform bacteria were not found on nonrinsed shredded cabbage on the initial day, injured coliforms at a range of 49-84% were detected on samples stored in air and high CO 2 atmospheres at 5°C and 10°C. Injured cells were detected more frequently during storage at both temperatures irrespective of the CO 2 atmosphere when shredded cabbage was rinsed with electrolyzed water. These results indicated that injured coliform bacteria on shredded cabbage, either rinsed or not rinsed with electrolyzed water, exhibited different degrees of injury during storage regardless of the CO 2 atmosphere and temperature tested. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Multiplexing spheroid volume, resazurin and acid phosphatase viability assays for high-throughput screening of tumour spheroids and stem cell neurospheres.

    Directory of Open Access Journals (Sweden)

    Delyan P Ivanov

    Full Text Available Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money.

  20. Multiplexing Spheroid Volume, Resazurin and Acid Phosphatase Viability Assays for High-Throughput Screening of Tumour Spheroids and Stem Cell Neurospheres

    Science.gov (United States)

    Ivanov, Delyan P.; Parker, Terry L.; Walker, David A.; Alexander, Cameron; Ashford, Marianne B.; Gellert, Paul R.; Garnett, Martin C.

    2014-01-01

    Three-dimensional cell culture has many advantages over monolayer cultures, and spheroids have been hailed as the best current representation of small avascular tumours in vitro. However their adoption in regular screening programs has been hindered by uneven culture growth, poor reproducibility and lack of high-throughput analysis methods for 3D. The objective of this study was to develop a method for a quick and reliable anticancer drug screen in 3D for tumour and human foetal brain tissue in order to investigate drug effectiveness and selective cytotoxic effects. Commercially available ultra-low attachment 96-well round-bottom plates were employed to culture spheroids in a rapid, reproducible manner amenable to automation. A set of three mechanistically different methods for spheroid health assessment (Spheroid volume, metabolic activity and acid phosphatase enzyme activity) were validated against cell numbers in healthy and drug-treated spheroids. An automated open-source ImageJ macro was developed to enable high-throughput volume measurements. Although spheroid volume determination was superior to the other assays, multiplexing it with resazurin reduction and phosphatase activity produced a richer picture of spheroid condition. The ability to distinguish between effects on malignant and the proliferating component of normal brain was tested using etoposide on UW228-3 medulloblastoma cell line and human neural stem cells. At levels below 10 µM etoposide exhibited higher toxicity towards proliferating stem cells, whereas at concentrations above 10 µM the tumour spheroids were affected to a greater extent. The high-throughput assay procedures use ready-made plates, open-source software and are compatible with standard plate readers, therefore offering high predictive power with substantial savings in time and money. PMID:25119185

  1. Evaluation of the thermotolerance of an inactivated rabies vaccine ...

    African Journals Online (AJOL)

    This study provides the first robust data that the antibody response of dogs vaccinated with Nobivac® Rabies vaccine stored for several months at high temperatures (up to 30 °C) is not inferior to that of dogs vaccinated with vaccine stored under recommended cold-chain conditions (2 - 8 °C). A controlled and randomized ...

  2. Induction of thermotolerance through heat acclimation in lablab ...

    African Journals Online (AJOL)

    The acclimation of plants to moderately high temperature plays an important role in inducing plant tolerance to subsequent lethal temperatures. This study was performed to investigate the effects of heat acclimation and sudden heat stress on the antioxidant and metabolic profile of lablab bean (Dolichos lablab). Following ...

  3. Validation of a PCR-based method for detection of food-borne thermotolerant Campylobacters in a multicenter collaborative trial

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Cook, N.; D'Agostino, M.

    2004-01-01

    A PCR-based method for rapid detection of food-borne thermotolerant campylobacters was evaluated through a collaborative trial with 12 laboratories testing spiked carcass rinse samples. The method showed an interlaboratory diagnostic sensitivity of 96.7% and a diagnostic specificity of 100...

  4. BACILLUS-METHANOLICUS SP-NOV, A NEW SPECIES OF THERMOTOLERANT, METHANOL-UTILIZING, ENDOSPORE-FORMING BACTERIA

    NARCIS (Netherlands)

    ARFMAN, N; DIJKHUIZEN, L; KIRCHHOF, G; LUDWIG, W; SCHLEIFER, KH; BULYGINA, ES; CHUMAKOV, KM; GOVORUKHINA, NI; TROTSENKO, YA; WHITE, D; SHARP, RJ

    The generic position of 14 strains of gram-positive bacteria able to use methanol as a growth substrate was determined. All are obligately aerobic, thermotolerant organisms that are able to grow at temperatures of 35 to 60-degrees-C. Nine of the strains produce oval spores at a

  5. Thermotolerance and heat stress responses of Douglas-fir and ponderosa pine seedling populations from contrasting climates

    Science.gov (United States)

    Danielle E. Marias; Frederick C. Meinzer; David R. Woodruff; Katherine A. McCulloh; David Tissue

    2016-01-01

    Temperature and the frequency and intensity of heat waves are predicted to increase throughout the 21st century. Germinant seedlings are expected to be particularly vulnerable to heat stress because they are in the boundary layer close to the soil surface where intense heating occurs in open habitats. We quantified leaf thermotolerance and whole-plant physiological...

  6. Genetically altering the expression of neutral trehalase gene affects conidiospore thermotolerance of the entomopathogenic fungus Metarhizium acridum

    Directory of Open Access Journals (Sweden)

    Peng Guoxiong

    2011-02-01

    Full Text Available Abstract Background The entomopathogenic fungus Metarhizium acridum has been used as an important biocontrol agent instead of insecticides for controlling crop pests throughout the world. However, its virulence varies with environmental factors, especially temperature. Neutral trehalase (Ntl hydrolyzes trehalose, which plays a role in environmental stress response in many organisms, including M. acridum. Demonstration of a relationship between Ntl and thermotolerance or virulence may offer a new strategy for enhancing conidiospore thermotolerance of entomopathogenic fungi through genetic engineering. Results We selected four Ntl over-expression and four Ntl RNA interference (RNAi transformations in which Ntl expression is different. Compared to the wild-type, Ntl mRNA expression was reduced to 35-66% in the RNAi mutants and increased by 2.5-3.5-fold in the over-expression mutants. The RNAi conidiospores exhibited less trehalase activity, accumulated more trehalose, and were much more tolerant of heat stress than the wild-type. The opposite effects were found in conidiospores of over-expression mutants compared to RNAi mutants. Furthermore, virulence was not altered in the two types of mutants compared to the wild type. Conclusions Ntl controlled trehalose accumulation in M. acridum by degrading trehalose, and thus affected conidiospore thermotolerance. These results offer a new strategy for enhancing conidiospore thermotolerance of entomopathogenic fungi without affecting virulence.

  7. Ectopic overexpression of SlHsfA3, a heat stress transcription factor from tomato, confers increased thermotolerance and salt hypersensitivity in germination in transgenic Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Zhenjun Li

    Full Text Available Plant heat stress transcription factors (Hsfs are the critical components involved in mediating responses to various environmental stressors. However, the detailed roles of many plant Hsfs are far from fully understood. In this study, an Hsf (SlHsfA3 was isolated from the cultivated tomato (Solanum lycopersicum, Sl and functionally characterized at the genetic and developmental levels. The nucleus-localized SlHsfA3 was basally and ubiquitously expressed in different plant organs. The expression of SlHsfA3 was induced dramatically by heat stress, moderately by high salinity, and slightly by drought, but was not induced by abscisic acid (ABA. The ectopic overexpression of SlHsfA3 conferred increased thermotolerance and late flowering phenotype to transgenic Arabidopsis plants. Moreover, SlHsfA3 played a negative role in controlling seed germination under salt stress. RNA-sequencing data demonstrated that a number of heat shock proteins (Hsps and stress-associated genes were induced in Arabidopsis plants overexpressing SlHsfA3. A gel shift experiment and transient expression assays in Nicotiana benthamiana leaves demonstrated that SlHsfA3 directly activates the expression of SlHsp26.1-P and SlHsp21.5-ER. Taken together, our results suggest that SlHsfA3 behaves as a typical Hsf to contribute to plant thermotolerance. The late flowering and seed germination phenotypes and the RNA-seq data derived from SlHsfA3 overexpression lines lend more credence to the hypothesis that plant Hsfs participate in diverse physiological and biochemical processes related to adverse conditions.

  8. Molecular basis for insecticide-enhanced thermotolerance in the brown planthopper Nilaparvata lugens Stål (Hemiptera:Delphacidae).

    Science.gov (United States)

    Ge, Lin-Quan; Huang, Liu-Juan; Yang, Guo-Qin; Song, Qi-Sheng; Stanley, David; Gurr, G M; Wu, Jin-Cai

    2013-11-01

    Climate change is likely to have marked ecological effects on terrestrial ecosystems, including the activities of insect pests. Most attention has focused on the increasing geographical ranges of pests; however, if extrinsic factors enhance their thermotolerance, populations may express increased voltinism and longer daily and annual activity periods. These changes in pest populations have the potential for severe consequences, including increased crop losses and decreased food security at the global level. The brown planthopper (BPH) Nilaparvata lugens Stål (Hemiptera: Delphacidae) is a serious pest of rice crops in temperate and tropical regions of Asia. It is often present in rice microclimates at temperatures close to its maximum thermotolerance. Recent BPH outbreaks in tropical Asia are considered to be associated with excess use of pesticides and increasing temperature. This study tested whether exposure to sublethal concentrations of triazophos (tzp), an insecticide widely used in Asian rice production, enhances thermotolerance of BPH. Tzp exposure (40 ppm at 40 °C) significantly decreased mortality (from 94% in controls to 50% at 48 h post-treatment) and increased lethal mean time (LT50 ) of adults by 17.2 h. To investigate the underlying molecular mechanism of this tzp-enhanced thermotolerance, we selected Hsp70 and Arginine kinase (Argk) for detailed study. Transcripts encoding both proteins in third-instar nymphs and brachypterous adult females were up-regulated, compared with controls, after exposure to tzp. RNAi silencing of both genes demonstrated that Hsp70 and Argk are essential for survival and tzp-increased thermotolerance. We propose that tzp induces thermotolerance in BPHs by increasing the expression of genes that act in cell protection mechanisms. The significance of our proposal relates to the importance of understanding the influence of sublethal concentrations of insecticides on pest biology. In addition to its influence on

  9. NADPH-thioredoxin reductase C mediates the response to oxidative stress and thermotolerance in the cyanobacterium Anabaena sp. PCC7120.

    Directory of Open Access Journals (Sweden)

    ANA MARÍA SÁNCHEZ-RIEGO

    2016-08-01

    Full Text Available NTRC (NADPH-thioredoxin reductase C is a bimodular enzyme composed of an NADPH-thioredoxin reductase and a thioredoxin domain extension in the same protein. In plants, NTRC has been described to be involved in the protection of the chloroplast against oxidative stress damage through reduction of the 2-Cys peroxiredoxin (2-Cys Prx as well as through other functions related to redox enzyme regulation. In cyanobacteria, the Anabaena NTRC has been characterized in vitro, however nothing was known about its in vivo function. In order to study that, we have generated the first knockout mutant strain (∆ntrC, apart from the previously described in Arabidopsis. Detailed characterization of this strain reveals a differential sensitivity to oxidative stress treatments with respect to the wild-type Anabaena strain, including a higher level of ROS (reactive oxygen species in normal growth conditions. In the mutant strain, different oxidative stress treatments such as hydrogen peroxide, methyl-viologen or high light irradiance provoke an increase in the expression of genes related to ROS detoxification, including AnNTRC and peroxiredoxin genes, with a concomitant increase in the amount of AnNTRC and 2-Cys Prx. Moreover, the role of AnNTRC in the antioxidant response is confirmed by the observation of a pronounced overoxidation of the 2-Cys Prx and a time-delay recovery of the reduced form of this protein upon oxidative stress treatments. Our results suggest the participation of this enzyme in the peroxide detoxification in Anabaena. In addition, we describe the role of Anabaena NTRC in thermotolerance, by the appearance of high molecular mass AnNTRC complexes, showing that the mutant strain is more sensitive to high temperature treatments.

  10. Acute effects of TiO2 nanomaterials on the viability and taxonomic composition of aquatic bacterial communities assessed via high-throughput screening and next generation sequencing.

    Directory of Open Access Journals (Sweden)

    Chu Thi Thanh Binh

    Full Text Available The nanotechnology industry is growing rapidly, leading to concerns about the potential ecological consequences of the release of engineered nanomaterials (ENMs to the environment. One challenge of assessing the ecological risks of ENMs is the incredible diversity of ENMs currently available and the rapid pace at which new ENMs are being developed. High-throughput screening (HTS is a popular approach to assessing ENM cytotoxicity that offers the opportunity to rapidly test in parallel a wide range of ENMs at multiple concentrations. However, current HTS approaches generally test one cell type at a time, which limits their ability to predict responses of complex microbial communities. In this study toxicity screening via a HTS platform was used in combination with next generation sequencing (NGS to assess responses of bacterial communities from two aquatic habitats, Lake Michigan (LM and the Chicago River (CR, to short-term exposure in their native waters to several commercial TiO2 nanomaterials under simulated solar irradiation. Results demonstrate that bacterial communities from LM and CR differed in their sensitivity to nano-TiO2, with the community from CR being more resistant. NGS analysis revealed that the composition of the bacterial communities from LM and CR were significantly altered by exposure to nano-TiO2, including decreases in overall bacterial diversity, decreases in the relative abundance of Actinomycetales, Sphingobacteriales, Limnohabitans, and Flavobacterium, and a significant increase in Limnobacter. These results suggest that the release of nano-TiO2 to the environment has the potential to alter the composition of aquatic bacterial communities, which could have implications for the stability and function of aquatic ecosystems. The novel combination of HTS and NGS described in this study represents a major advance over current methods for assessing ENM ecotoxicity because the relative toxicities of multiple ENMs to thousands

  11. Mg(2+) improves the thermotolerance of probiotic Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8.

    Science.gov (United States)

    Yang, Y; Huang, S; Wang, J; Jan, G; Jeantet, R; Chen, X D

    2017-04-01

    Food-related carbohydrates and proteins are often used as thermoprotectants for probiotic lactobacilli during industrial production and processing. However, the effect of inorganic salts is rarely reported. Magnesium is the second-most abundant cation in bacteria, and commonly found in various foods. Mg(2+) homeostasis is important in Salmonella and has been reported to play a critical role in their thermotolerance. However, the role of Mg(2+) in thermotolerance of other bacteria, in particular probiotic bacteria, still remains a hypothesis. In this study, the effect of Mg(2+) on thermotolerance of probiotic lactobacilli was investigated in three well-documented probiotic strains, Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8, in comparison with Zn(2+) and Na(+) . Concentrations of Mg(2+) between 10 and 50 mmol l(-1) were found to increase the bacterial survival upon heat challenge. Remarkably, Mg(2+) addition at 20 mmol l(-1) led to a 100-fold higher survival of L. rhamnosus GG upon heat challenge. This preliminary study also showed that Mg(2+) shortened the heat-induced extended lag time of bacteria, which indicated the improvement in bacterial recovery from thermal injury. In order to improve the productivity and stability of live probiotics, extensive investigations have been carried out to improve thermotolerance of probiotics. However, most of these studies focused on the effects of carbohydrates, proteins or amino acids. The roles of inorganic salts in various food materials, which have rarely been reported, should be considered when incorporating probiotics into these foods. In this study, Mg(2+) was found to play a significant role in the thermotolerance of probiotic lactobacilli. A novel strategy may be available in the near future by employing magnesium salts as protective agents of probiotics during manufacturing process. © 2017 The Society for Applied Microbiology.

  12. Bacterial metabolites from intra- and inter-species influencing thermotolerance: the case of Bacillus cereus and Geobacillus stearothermophilus.

    Science.gov (United States)

    Gómez-Govea, Mayra Alejandra; García, Santos; Heredia, Norma

    2017-05-01

    Bacterial metabolites with communicative functions could provide protection against stress conditions to members of the same species. Yet, information remains limited about protection provided by metabolites in Bacillus cereus and inter-species. This study investigated the effect of extracellular compounds derived from heat shocked (HS) and non-HS cultures of B. cereus and Geobacillus stearothermophilus on the thermotolerance of non-HS vegetative and sporulating B. cereus. Cultures of B. cereus and G. stearothermophilus were subjected to HS (42 or 65 °C respectively for 30 min) or non-HS treatments. Cells and supernatants were separated, mixed in a combined array, and then exposed to 50 °C for 60 min and viable cells determined. For spores, D values (85 and 95 °C) were evaluated after 120 h. In most cases, supernatants from HS B. cereus cultures added to non-HS B. cereus cells caused their thermotolerance to increase (D 50 12.2-51.9) when compared to supernatants from non-HS cultures (D 50 7.4-21.7). While the addition of supernatants from HS and non-HS G. stearothermophilus cultures caused the thermotolerance of non-HS cells from B. cereus to decrease initially (D 50 3.7-7.1), a subsequent increase was detected in most cases (D 50 18-97.7). In most cases, supernatants from sporulating G. stearothermophilus added to sporulating cells of B. cereus caused the thermotolerance of B. cereus 4810 spores to decline, whereas that of B. cereus 14579 increased. This study clearly shows that metabolites in supernatants from either the same or different species (such as G. stearothermophilus) influence the thermotolerance of B. cereus.

  13. Abundance, viability and culturability of Antarctic bacteria

    Digital Repository Service at National Institute of Oceanography (India)

    LokaBharathi, P.A.; DeSouza, M.J.B.D.; Nair, S.; Chandramohan, D.

    The viability of total number of bacteria decide the mineralisation rate in any ecosystem and ultimately the fertility of the region. This study aims at establishing the extent of viability in the standing stock of the Antarctic bacterial population...

  14. STUDY ON POLLEN VIABILITY AS BIOINDICATOR OF AIR QUALITY

    Directory of Open Access Journals (Sweden)

    Florentina ŞTEFLEA

    2012-01-01

    Full Text Available The aim of this research is to estimate the relationship between pollen viability and atmospheric pollution (in polluted and non-polluted conditions. The study was carried out in the city of Timisoara. Two areas, with different intensity of road traffic (very high and absent but all characterized by the presence of the same plant species, were selected. The pollen of herbaceous spontaneous species, arboreal species and a shrub species was used (Robinia pseudacacia, Aesculus x carnea, Catalpa bignonioides, Albizzia julibrissin, Rosa canina, Sambucus nigra, Malva neglecta, Ranunculus acer, Trifolium repens, Cichorium intybus. The pollen of these species was treated with TTC (2, 3, 5 Tryphenil-Tetrazolium-Chloride staining solution and viability was then estimated by light microscopy. The results of the mean pollen viability percentage of the examined species are reported. Pollen viability of herbaceous plants is significantly different between the two environments.

  15. Deficiencies in mitochondrial DNA compromise the survival of yeast cells at critically high temperatures.

    Science.gov (United States)

    Zubko, Elena I; Zubko, Mikhajlo K

    2014-01-01

    To address possible roles of mitochondrial genes in adaptation of eukaryotic cells to critical temperatures, we compared thermotolerance of mitochondrial rho mutants and wild type cells of six rho positive yeast species: Candida glabrata, Saccharomyces bayanus, Saccharomyces cerevisiae, Saccharomyces eubayanus, Saccharomyces paradoxus and Saccharomyces pastorianus. All rho mutants manifested compromised thermotolerance as a common phenotype. Analysis of viabilities at critical temperatures (32-45 °C) showed the reduction of maximum permissive temperatures (MPTs) in all rho mutants in comparison to their wild type counterparts. Degrees of the compromised thermotolerance depended on maximum permissive temperatures for wild type cells: the highest levels of MPT reductions for rho mutants took place in species and strains with highest MPTs for wild types. Short term exposures of S. cerevisiae cells (up to 3.5h) at non-permissive temperatures (45 °C and 50 °C) also lead to more rapid cell death of rho mutants as compared to wild type cells. We conclude that: (1) compromised thermotolerance could be a generic phenotypic property of rho mutants; (2) the enhanced thermotolerance of cells possessing mitochondrial genomes could be one of selective advantages in adaptation to environmental factors, in particular to enhanced temperatures. Copyright © 2013 Elsevier GmbH. All rights reserved.

  16. Tychastic measure of viability risk

    CERN Document Server

    Aubin, Jean-Pierre; Dordan, Olivier

    2014-01-01

    This book presents a forecasting mechanism of the price intervals for deriving the SCR (solvency capital requirement) eradicating the risk during the exercise period on one hand, and measuring the risk by computing the hedging exit time function associating with smaller investments the date until which the value of the portfolio hedges the liabilities on the other. This information, summarized under the term “tychastic viability measure of risk” is an evolutionary alternative to statistical measures, when dealing with evolutions under uncertainty. The book is written by experts in the field and the target audience primarily comprises research experts and practitioners.

  17. Cyclosporin A treatment of Leishmania donovani reveals stage-specific functions of cyclophilins in parasite proliferation and viability.

    Directory of Open Access Journals (Sweden)

    Wai-Lok Yau

    Full Text Available BACKGROUND: Cyclosporin A (CsA has important anti-microbial activity against parasites of the genus Leishmania, suggesting CsA-binding cyclophilins (CyPs as potential drug targets. However, no information is available on the genetic diversity of this important protein family, and the mechanisms underlying the cytotoxic effects of CsA on intracellular amastigotes are only poorly understood. Here, we performed a first genome-wide analysis of Leishmania CyPs and investigated the effects of CsA on host-free L. donovani amastigotes in order to elucidate the relevance of these parasite proteins for drug development. METHODOLOGY/PRINCIPAL FINDINGS: Multiple sequence alignment and cluster analysis identified 17 Leishmania CyPs with significant sequence differences to human CyPs, but with highly conserved functional residues implicated in PPIase function and CsA binding. CsA treatment of promastigotes resulted in a dose-dependent inhibition of cell growth with an IC50 between 15 and 20 microM as demonstrated by proliferation assay and cell cycle analysis. Scanning electron microscopy revealed striking morphological changes in CsA treated promastigotes reminiscent to developing amastigotes, suggesting a role for parasite CyPs in Leishmania differentiation. In contrast to promastigotes, CsA was highly toxic to amastigotes with an IC50 between 5 and 10 microM, revealing for the first time a direct lethal effect of CsA on the pathogenic mammalian stage linked to parasite thermotolerance, independent from host CyPs. Structural modeling, enrichment of CsA-binding proteins from parasite extracts by FPLC, and PPIase activity assays revealed direct interaction of the inhibitor with LmaCyP40, a bifunctional cyclophilin with potential co-chaperone function. CONCLUSIONS/SIGNIFICANCE: The evolutionary expansion of the Leishmania CyP protein family and the toxicity of CsA on host-free amastigotes suggest important roles of PPIases in parasite biology and implicate

  18. ADPase activity of recombinantly expressed thermotolerant ATPases may be caused by copurification of adenylate kinase of Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Baoyu; Sysoeva, Tatyana A.; Chowdhury, Saikat; Guo, Liang; Nixon, B.Tracy; (IIT); (Penn)

    2009-10-06

    Except for apyrases, ATPases generally target only the {gamma}-phosphate of a nucleotide. Some non-apyrase ATPases from thermophilic microorganisms are reported to hydrolyze ADP as well as ATP, which has been described as a novel property of the ATPases from extreme thermophiles. Here, we describe an apparent ADP hydrolysis by highly purified preparations of the AAA+ ATPase NtrC1 from an extremely thermophilic bacterium, Aquifex aeolicus. This activity is actually a combination of the activities of the ATPase and contaminating adenylate kinase (AK) from Escherichia coli, which is present at 1/10 000 of the level of the ATPase. AK catalyzes conversion of two molecules of ADP into AMP and ATP, the latter being a substrate for the ATPase. We raise concern that the observed thermotolerance of E. coli AK and its copurification with thermostable proteins by commonly used methods may confound studies of enzymes that specifically catalyze hydrolysis of nucleoside diphosphates or triphosphates. For example, contamination with E. coli AK may be responsible for reported ADPase activities of the ATPase chaperonins from Pyrococcus furiosus, Pyrococcus horikoshii, Methanococcus jannaschii and Thermoplasma acidophilum; the ATP/ADP-dependent DNA ligases from Aeropyrum pernix K1 and Staphylothermus marinus; or the reported ATP-dependent activities of ADP-dependent phosphofructokinase of P. furiosus. Purification methods developed to separate NtrC1 ATPase from AK also revealed two distinct forms of the ATPase. One is tightly bound to ADP or GDP and able to bind to Q but not S ion exchange matrixes. The other is nucleotide-free and binds to both Q and S ion exchange matrixes.

  19. Viability of pollen grains of tetraploid banana

    Directory of Open Access Journals (Sweden)

    Taliane Leila Soares

    2016-01-01

    Full Text Available ABSTRACT Obtaining banana tetraploid cultivars from triploid strains results in total or partial reestablishment of fertility, allowing the occurrence of some fruits with seeds, a feature that is undesirable from a marketing perspective. The objective of this study was to assess the viability of pollen of 12 banana tetraploid hybrids (AAAB by means of in vitro germination and two histochemical tests (acetocarmine and 2,3,5-triphenyltetrazolium chloride. The pollen tube growth was evaluated by germinating grains in three culture media — M1: 0.03% Ca(NO3∙4H2O, 0.02% Mg(SO4∙7H2O, 0.01% KNO3, 0.01% H3BO3 and 15% sucrose; M2: 0.03% Ca(NO3∙4H2O, 0.01% KNO3, 0.01% H3BO3 and 10% sucrose; and M3: 0.015% H3BO3, 0.045% Ca3(PO42 and 25% sucrose. The acetocarmine staining indicated high viability (above 80%, except for the genotypes YB42-17 and Caprichosa, which were 76 and 70%, respectively. However, the in vitro germination rate was lower than 50% for all the genotypes, except for the hybrids YB42-17 (M1 and YB42-47 (M1. The medium M1 provided the greatest germination percentage and pollen tube growth. Among the genotypes assessed, YB42-47 presented the highest germination rate (61.5% and tube length (5.0 mm. On the other hand, the Vitória cultivar had the lowest germination percentage (8.2% in medium M1. Studies of meiosis can shed more light on the differences observed in the evaluated tetraploids, since meiotic irregularities can affect pollen viability.

  20. Impact of urban contamination of the La Paz River basin on thermotolerant coliform density and occurrence of multiple antibiotic resistant enteric pathogens in river water, irrigated soil and fresh vegetables.

    Science.gov (United States)

    Poma, Violeta; Mamani, Nataniel; Iñiguez, Volga

    2016-01-01

    present. Moreover, some Salmonella isolates were distinguished by their multi-resistance to ≥8 antibiotics, within soil and vegetable samples. Overall, this study demonstrates that La Paz River-an affluent of the Amazon macrobasin-is heavily polluted along the year with a high density of thermotolerant coliforms and is a reservoir of multiple antibiotic resistant enteropathogens, present in river water, soil and vegetables. These data highlight health risk associated with food and waterborne diseases at the region.

  1. Prevalence and Antibiotic Resistance of Thermotolerant Campylobacter spp. in Chicken Meat Sold in İstanbul

    Directory of Open Access Journals (Sweden)

    Serkan Kemal BÜYÜKÜNAL

    2017-07-01

    Full Text Available Campylobacter spp. are some of the most common causes of bacterial diarrhea in humans worldwide. They are mainly considered as foodborne pathogens that are found in raw or undercooked poultry and serve as an important source of sporadic campylobacteriosis. The present study was aimed to determine the prevalence and the antimicrobial resistance patterns of thermotolerant Campylobacter spp. in chicken meat. A total of 176 samples of chicken meat were analyzed using PCM and BAX® system. The samples analyzed included: 56 samples of whole chicken, 27 samples of chicken breast, 33 samples of chicken thigh, 25 samples of chicken drumstick and 35 samples of chicken wings. Samples of all the fresh chicken meat sold in İstanbul were randomly purchased from different major supermarkets in their original, individual packages. Laboratory analyses to detect thermotolerant Campylobacter spp. were performed in accordance with the ISO 10272-1, 2006 standard (qualitative analysis. API® Campy (BioMerieux, Marcy-l’Etoile, France was used for the confirmation of presumptive colonies. Campylobacter isolates were subjected to antimicrobial susceptibility tests by the disc diffusion method as recommended by the National Committee for Clinical Laboratory Standards. Zones of growth inhibition were evaluated according to the NCCLS standards. Using PCM, the prevalence of C. coli, C. jejuni and C. lari was determined as 15.34, 8.52 and 1.7%, respectively. However, using BAX® system, the prevalence was determined as 15.90, 18.75 and 1.7% for C. coli, C. jejuni and C. lari, respectively. C. coli was resistant to nalidixic acid (78.57%, ofloxacin (14.29% norfloxacin (10.71% and ampicillin (10.71%. But the highest resistance was observed to nalidixic acid (90.91% for C. jejuni and (100% for C. lari. In conclusion, considering the public health, chicken meat is a common source for Campylobacter strains and antibiotics should be used carefully in veterinary medicine.

  2. Screening of thermotolerant microorganisms and application for oil separation from palm oil mill wastewater

    Directory of Open Access Journals (Sweden)

    Aran H-Kittikun

    2007-05-01

    Full Text Available The characteristics of palm oil mill wastewater (POMW were brown color, pH 3.8-4.3, temperature 48-55oC, total solids 68.2-82.1 g/l, suspended solids 26.2-65.6 g/l, oil and grease 19.1-25.1 g/l, COD 49.9-160.7g/l and BOD 32.5-75.3 g/l. After centrifugation (3,184 xg of 50 ml POMW for 10 min, the POMW was separated into 3 layers: top (oil, middle (supernatant and bottom layer (sediment. The sediment containeddry weight 1.19 g and oil and grease 1.07 g. In order to release oil and grease trapped in palm fiber debris in the POMW, cellulase- and/or xylanase-enzyme-producing and thermotolerant microorganisms wereisolated. The isolates SO1 and SO2 were isolated from soil near the first anaerobic pond of the palm oil mill. They were aerobic, Gram positive, rod shaped, thermotolerant microorganisms and produced cellulase 12.11 U/ml (3 days and 7.2 U/ml (4 days, and xylanase 50.98 U/ml (4 days and 20.42 U/ml (4 days, respectivelyin synthetic medium containing carboxymethycellulose as a carbon source. When these 2 isolates were added into the steriled POMW under shaking condition for 7 days, after centrifugation at 3,184 xg the isolate SO1gave the better % reduction of dry weight (64.66 % and of oil and grease in the bottom layer (85.32 % of the POMW.

  3. Role of redox homeostasis in thermo-tolerance under a climate change scenario.

    Science.gov (United States)

    de Pinto, Maria Concetta; Locato, Vittoria; Paradiso, Annalisa; De Gara, Laura

    2015-09-01

    Climate change predictions indicate a progressive increase in average temperatures and an increase in the frequency of heatwaves, which will have a negative impact on crop productivity. Over the last decade, a number of studies have addressed the question of how model plants or specific crops modify their metabolism when exposed to heat stress. This review provides an overview of the redox pathways that contribute to how plants cope with heat stress. The focus is on the role of reactive oxygen species (ROS), redox metabolites and enzymes in the signalling pathways leading to the activation of defence responses. Additional attention is paid to the regulating mechanisms that lead to an increase in specific ROS-scavenging systems during heat stress, which have been studied in different model systems. Finally, increasing thermo-tolerance in model and crop plants by exposing them to heat acclimation or to exogenous treatments is discussed. Although there is clear evidence that several strategies are specifically activated according to the intensity and the duration of heat stress, as well as the capacity of the different species or genotypes to overcome stress, an alteration in redox homeostasis seems to be a common event. Different mechanisms that act to enhance redox systems enable crops to overcome heat stress more effectively. Knowledge of thermo-tolerance within agronomic biodiversity is thus of key importance to enable researchers to identify new strategies for overcoming the impacts of climate change, and for decision-makers in planning for an uncertain future with new choices and options open to them. © The Author 2015. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  4. Genetic engineering of the biosynthesis of glycinebetaine enhances thermotolerance of photosystem II in tobacco plants.

    Science.gov (United States)

    Yang, Xinghong; Wen, Xiaogang; Gong, Hongmei; Lu, Qingtao; Yang, Zhipan; Tang, Yunlai; Liang, Zheng; Lu, Congming

    2007-02-01

    Genetically engineered tobacco (Nicotiana tabacum L.) with the ability to accumulate glycinebetaine was established. The wild type and transgenic plants were exposed to heat treatment (25-50 degrees C) for 4 h in the dark and under growth light intensity (300 mumol m(-2) s(-1)). The analyses of oxygen-evolving activity and chlorophyll fluorescence demonstrated that photosystem II (PSII) in transgenic plants showed higher thermotolerance than in wild type plants in particular when heat stress was performed in the light, suggesting that the accumulation of glycinebetaine leads to increased tolerance to heat-enhanced photoinhibition. This increased tolerance was associated with an improvement on thermostability of the oxygen-evolving complex and the reaction center of PSII. The enhanced tolerance was caused by acceleration of the repair of PSII from heat-enhanced photoinhibition. Under heat stress, there was a significant accumulation of H(2)O(2), O (2) (-) and catalytic Fe in wild type plants but this accumulation was much less in transgenic plants. Heat stress significantly decreased the activities of catalase, ascorbate peroxidase, glutathione reductase, dehydroascorbate reductase, and monodehydroascorbate reductase in wild type plants whereas the activities of these enzymes either decreased much less or maintained or even increased in transgenic plants. In addition, heat stress increased the activity of superoxide dismutase in wild type plants but this increase was much greater in transgenic plants. Furthermore, transgenic plants also showed higher content of ascorbate and reduced glutathione than that of wild type plants under heat stress. The results suggest that the increased thermotolerance induced by accumulation of glycinebetaine in vivo was associated with the enhancement of the repair of PSII from heat-enhanced photo inhibition, which might be due to less accumulation of reactive oxygen species in transgenic plants.

  5. Depth of morphologic skin damage and viability after one, two, and three passes of a high-energy, short-pulse CO2 laser (Tru-Pulse) in pig skin.

    Science.gov (United States)

    Smith, K J; Skelton, H G; Graham, J S; Hamilton, T A; Hackley, B E; Hurst, C G

    1997-08-01

    CO2 laser energy is absorbed by water, which is present in all tissue. The depth of penetration of CO2 lasers is narrow with minimal reflection, scatter, or transmission. However, thermal damage has limited the usefulness of conventional, continuous-wave CO2 lasers for debridement as demonstrated by wound healing studies. The development of high-energy CO2 lasers, with pulse durations that are less than the thermal relaxation time of tissue, have made vaporization of skin for resurfacing and wound debridement possible because of the decreased risk of thermal damage. This study was performed to evaluate thermal damage produced by a CO2 laser. Routine histopathologic examination and nitroblue-tetrazolium chloride (NBTC) staining were used to evaluate the depth of tissue damage and viability in weanling pig skin after one, two, and three passes of the laser. At a pulse energy of 300 mJ, with a pulse duration of 60 microseconds, one pass of the laser produced vaporization of the epidermis with minimal thermal damage. Two passes produced areas of denatured collagen with loss of viable cells in the superficial papillary dermis. Three passes extended the damage into the papillary dermis. Hyalinization of collagen appears to correspond well with the level of thermal damage as measured by NBTC staining. Our findings suggest that the energy necessary to vaporize the dermis may be greater than that needed to vaporize epidermis.

  6. Confirmation of E. coli among other thermotolerant coliform bacteria in paper mill effluents, wood chips screening rejects and paper sludges.

    Science.gov (United States)

    Beauchamp, Chantal J; Simao-Beaunoir, Anne-Marie; Beaulieu, Carole; Chalifour, François-Philippe

    2006-07-01

    Paper sludges are solid wastes material generated from the paper production, which have been characterized for their chemical contents. Some are rich in wood fiber and are a good carbon source, for example the primary and de-inking paper sludges. Others are made rich in nitrogen and phosphorus by pressing the activated sludge, resulting from the biological water treatments, with the primary sludge, yielding the combined paper sludge. Still, in the absence of sanitary effluents very few studies have addressed the characterization of their coliform microflora. Therefore, this study investigated the thermotolerant coliform population of one paper mill effluent and two paper mill sludges and wood chips screening rejects using chromogenic media. For the first series of analyses, the medium used was Colilert broth and positive tubes were selected to isolate bacteria in pure culture on MacConkey agar. In a second series of analyses, double selective media, based on ss-galactosidase and ss-glucuronidase activities, were used to isolate bacteria. First, the presence of thermotolerant coliforms was detected in low numbers in most water effluents, but showed that the entrance of the thermotolerant coliforms was early in the industrial process. Also, large numbers of thermotolerant coliforms, i.e., 7,000,000 MPN/g sludge (dry weight; d.w.), were found in combined sludges. From this first series of isolations, bacteria were purified on MacConkey medium and identified as Citrobacter freundii, Enterobacter sp, E. sakazakii, E. cloacae, Escherichia coli, K. pneumoniae, K. pneumoniae subsp. rhinoscleromatis, K. pneumoniae subsp. ozaenae, K. pneumoniae subsp. pneumoniae, Pantoea sp, Raoultella terrigena, R. planticola. Second, the presence of thermotolerant coliforms was measured at more than 3,700-6,000 MPN/g (d.w) sludge, whereas E. coli was detected from 730 to more than 3,300 MPN/g (d.w.) sludge. The presence of thermotolerant coliform bacteria and E. coli was sometimes detected

  7. Wildlife Tunnel Enhances Population Viability

    Directory of Open Access Journals (Sweden)

    Rodney van der Ree

    2009-12-01

    Full Text Available Roads and traffic are pervasive components of landscapes throughout the world: they cause wildlife mortality, disrupt animal movements, and increase the risk of extinction. Expensive engineering solutions, such as overpasses and tunnels, are increasingly being adopted to mitigate these effects. Although some species readily use such structures, their success in preventing population extinction remains unknown. Here, we use population viability modeling to assess the effectiveness of tunnels for the endangered Mountain Pygmy-possum (Burramys parvus in Australia. The underpasses reduced, but did not completely remove, the negative effects of a road. The expected minimum population size of a "reconnected" population remained 15% lower than that of a comparable "undivided" population. We propose that the extent to which the risk of extinction decreases should be adopted as a measure of effectiveness of mitigation measures and that the use of population modeling become routine in these evaluations.

  8. Malthus, Boserup and population viability.

    Science.gov (United States)

    Bonneuil, N

    1994-01-01

    The Malthus-Boserup explanatory framework is revisited from the point of view of viability theory. Instead of imposing a univocal relationship between population pressure and level of knowledge, the way technology will change is not determined, it is only constrained. This leads to regard any situation as associated to a set of reachable futures. When no possibility is left for systems to avoid extinction, systems are no longer viable. Hence, the control-phase space can be divided into regions corresponding to gradual danger or security. This point of view allows the introduction of ideas such as incentives to create or to use new knowledge, gives a role to the threatening power of Malthusian checks, and leaves space for a specific variety of behaviors. The Boserupian theme then appears indirectly, emerging from the constraints imposed by the inertia of technological change.

  9. Viability of telework at PROCEMPA.

    Science.gov (United States)

    Fetzner, Maria Amelia de Mesquita

    2003-02-01

    At the end of the 20th century, telework appears as one of the modalities of flexible work, which is related to new organizational structures as well as to increasing use of technology. It revolutionizes the traditional ways of performing work. Its implementation creates a number of questions to be answered by the organizations and the individuals involved. This article presents a case study on the viability of implementing telework at Procempa (The Data Processing Company of the City of Porto Alegre). The case study analyzes the technical, organizational, psychological, legal, and labor union dimensions. As a result of this study, we can identify the organization's stage of readiness for telework, the conditions under which it would be implemented, and the specific issues of an implementation.

  10. BIOSYNTHESIS GLUCOAMYLASE, AMINO ACIDS, FATTY ACIDS, LIPIDS THERMOTOLERANT AND MESOPHILIC STRAINS OF THE GENUS ASPERGILLUS AT DIFFERENT TEMPERATURES

    Directory of Open Access Journals (Sweden)

    V. S. Grigorov

    2014-01-01

    Full Text Available Summary. Active enzyme synthesis micromycetes thermotolerance at elevated temperatures incentives presence of acidic amino acids in the medium. In mesophilic micromycetes increased content of unsaturated fatty acids and reduced enzyme activity The effect of elevated temperature on the biosynthesis of glucoamylases and quantitative changes of some low molecular weight components in cells Aspergillus fungi. The object of the Exploration were taken micromycetes Aspergillus awamori WUD-T-2 and Aspergillus awamori Czech strain. Strains were cultivated for 96 hours at temperatures of 30, 40, 45 ° C in 750 cm3 flasks on shuttel - unit with a frequency of rotation of the 4-to-1. It has been established that the increase of the temperature of cultivation thermotolerance micromycetes from 30 to 45 ° C in those chenie 96 h increases intracellular content of amino acids, especially glutamic and aspartic and mono acyl glyceride and polar lipids, triglycerides of fatty acids having an even number of carbon atoms. Increasing the temperature of cultivation thermotolerance strain helped to increase the content of these lipid fractions; at mesophilic strain their content decreased at 45 oC. In the mycelium of the fungus Aspergillus awamori WUD-T-2, an increase in temperature causes an increase in the synthesis of palmitic and stearic acids, linoleic and linoene decreases. From a strain of Aspergillus awamori stкфшт Czech amounts of saturated fatty acids has changed little, is unsaturated - is reduced. Glucoamylase activity in the mycelium and culture fluid thermotolerance achievement-a maximum at 45 ° C, in the mesophile - at 40 oC. Thus, at supraoptimal temperatures culturing Aspergillus awamori WUD-T-2 is enhanced metabolic processes producing by hydrophobic interactions and s-amino acids with the ionized-COOH groups increased content of saturated fatty acids, thereby increasing biosynthesis glucoamylase, which was not observed in mesophyll strain.

  11. Calcium/calmodulin-dependent kinases are involved in growth, thermotolerance, oxidative stress survival, and fertility in Neurospora crassa.

    Science.gov (United States)

    Kumar, Ravi; Tamuli, Ranjan

    2014-04-01

    Calcium/calmodulin-dependent kinases (Ca(2+)/CaMKs) are Ser/Thr protein kinases that respond to change in cytosolic free Ca(2+) ([Ca(2+)]c) and play multiple cellular roles in organisms ranging from fungi to humans. In the filamentous fungus Neurospora crassa, four Ca(2+)/CaM-dependent kinases, Ca(2+)/CaMK-1 to 4, are encoded by the genes NCU09123, NCU02283, NCU06177, and NCU09212, respectively. We found that camk-1 and camk-2 are essential for full fertility in N. crassa. The survival of ∆camk-2 mutant was increased in induced thermotolerance and oxidative stress conditions. In addition, the ∆camk-1 ∆camk-2, ∆camk-4 ∆camk-2, and ∆camk-3 ∆camk-2 double mutants display slow growth phenotype, reduced aerial hyphae, decreased thermotolerance, and increased sensitivity to oxidative stress, revealing the genetic interactions among these kinases. Therefore, Ca(2+)/CaMKs are involved in growth, thermotolerance, oxidative stress tolerance, and fertility in N. crassa.

  12. Aerosol dissemination veterinary pathogenic and human opportunistic thermophilic and thermotolerant fungi from thermal effluents of nuclear production reactors

    Energy Technology Data Exchange (ETDEWEB)

    Tansey, M.R.; Fliermans, C.B.; Kern, C.D.

    1979-01-01

    The extent to which veterinary pathogenic and human opportunistic species of thermophilic and thermotolerant fungi disseminate in aerosols from heated effluents of nuclear production reactors of the Savannah River Plant (SRP), South Carolina, has been measured. Aerosol samples were taken at 140 sites, from directly over thermal effluents to more than 100 kilometers from the SRP boundary. Sampling methods included settle plates, liquid impingement, filtration, and a particle sizing cascade impactor (Andersen Sampler). Soils, foams, and microbial mats from thermal effluents, and vegetation were sampled to study distribution of particular species. Sampling was done under a variety of conditions; hot weather and cold, wet and dry, day and night, windy and calm, reactor(s) operating and not, disturbed vegetation and undisturbed. At 102 of the aerosol sampling sites, sophisticated meterological analysis were used to allow sampling of air in the plume which originated from thermal effluents. Soil, foam, microbial mat, vegetation, and aerosol samples were quantitatively plated for detection of viable units; filters were halved and then both plated and observed microscopically. Significant dissemination of thermophilic and thermotolerant fungi from thermal effluents was not detected. Thermophilic and thermotolerant fungi were widely distributed in soil, air, and on vegetation. Dactylaria gallopava, the indicator species and dominant fungus in microbial mats lining SRP thermal effluents and the cause of epidemic fatal phaeohyphomycosis in flocks of turkeys and chickens in South Carolina, Georgia, and elsewhere, was isolated from air at a maximum of 50 meters from effluents.

  13. Relationship between humidity and influenza A viability in droplets and implications for influenza's seasonality.

    Directory of Open Access Journals (Sweden)

    Wan Yang

    Full Text Available Humidity has been associated with influenza's seasonality, but the mechanisms underlying the relationship remain unclear. There is no consistent explanation for influenza's transmission patterns that applies to both temperate and tropical regions. This study aimed to determine the relationship between ambient humidity and viability of the influenza A virus (IAV during transmission between hosts and to explain the mechanisms underlying it. We measured the viability of IAV in droplets consisting of various model media, chosen to isolate effects of salts and proteins found in respiratory fluid, and in human mucus, at relative humidities (RH ranging from 17% to 100%. In all media and mucus, viability was highest when RH was either close to 100% or below ∼50%. When RH decreased from 84% to 50%, the relationship between viability and RH depended on droplet composition: viability decreased in saline solutions, did not change significantly in solutions supplemented with proteins, and increased dramatically in mucus. Additionally, viral decay increased linearly with salt concentration in saline solutions but not when they were supplemented with proteins. There appear to be three regimes of IAV viability in droplets, defined by humidity: physiological conditions (∼100% RH with high viability, concentrated conditions (50% to near 100% RH with lower viability depending on the composition of media, and dry conditions (<50% RH with high viability. This paradigm could help resolve conflicting findings in the literature on the relationship between IAV viability in aerosols and humidity, and results in human mucus could help explain influenza's seasonality in different regions.

  14. Economic viability of anaerobic digestion

    Energy Technology Data Exchange (ETDEWEB)

    Wellinger, A. [INFOENERGIE, Ettenhausen (Switzerland)

    1996-01-01

    The industrial application of anaerobic digestion is a relatively new, yet proven waste treatment technology. Anaerobic digestion reduces and upgrades organic waste, and is a good way to control air pollution as it reduces methane and nitrous gas emissions. For environmental and energy considerations, anaerobic digestion is a nearly perfect waste treatment process. However, its economic viability is still in question. A number of parameters - type of waste (solid or liquid), digester system, facility size, product quality and end use, environmental requirements, cost of alternative treatments (including labor), and interest rates - define the investment and operating costs of an anaerobic digestion facility. Therefore, identical facilities that treat the same amount and type of waste may, depending on location, legislation, and end product characteristics, reveal radically different costs. A good approach for evaluating the economics of anaerobic digestion is to compare it to treatment techniques such as aeration or conventional sewage treatment (for industrial wastewater), or composting and incineration (for solid organic waste). For example, the cost (per ton of waste) of in-vessel composting with biofilters is somewhat higher than that of anaerobic digestion, but the investment costs 1 1/2 to 2 times more than either composting or anaerobic digestion. Two distinct advantages of anaerobic digestion are: (1) it requires less land than either composting or incinerating, which translates into lower costs and milder environmental and community impacts (especially in densely populated areas); and (2) it produces net energy, which can be used to operate the facility or sold to nearby industries.

  15. Evaluation of Fermentation, Drying, and/or High Pressure Processing on Viability of Listeria monocytogenes, Escherichia coli O157:H7, Salmonella spp., and Trichinella spiralis in Raw Pork and Genoa Salami

    Science.gov (United States)

    In the present study, we evaluated the effect of fermentation/drying on viability of T. spiralis in Genoa salami. We also evaluated HPP for efficacy towards T. spiralis larvae in trichinae-infected pig masseter muscle as an alternate to curing for trichinae control. We also validated the integrated ...

  16. A thermotolerant beta-glucosidase isolated from an endophytic fungi, Periconia sp., with a possible use for biomass conversion to sugars.

    Science.gov (United States)

    Harnpicharnchai, Piyanun; Champreda, Verawat; Sornlake, Warasirin; Eurwilaichitr, Lily

    2009-10-01

    A fungal strain, BCC2871 (Periconia sp.), was found to produce a thermotolerant beta-glucosidase, BGL I, with high potential for application in biomass conversion. The full-length gene encoding the target enzyme was identified and cloned into Pichia pastoris KM71. Similar to the native enzyme produced by BCC2871, the recombinant beta-glucosidase showed optimal temperature at 70 degrees C and optimal pH of 5 and 6. The enzyme continued to exhibit high activity even after long incubation at high temperature, retaining almost 60% of maximal activity after 1.5h at 70 degrees C. It was also stable under basic conditions, retaining almost 100% of maximal activity after incubation for 2h at pH8. The enzyme has high activity towards cellobiose and other synthetic substrates containing glycosyl groups as well as cellulosic activity toward carboxymethylcellulose. Thermostability of the enzyme was improved remarkably in the presence of cellobiose, glucose, or sucrose. This beta-glucosidase was able to hydrolyze rice straw into simple sugars. The addition of this beta-glucosidase to the rice straw hydrolysis reaction containing a commercial cellulase, Celluclast 1.5L (Novozyme, Denmark) resulted in increase of reducing sugars being released compared to the hydrolysis without the beta-glucosidase. This enzyme is a candidate for applications that convert lignocellulosic biomass to biofuels and chemicals.

  17. Multimodality imaging in the assessment of myocardial viability

    Science.gov (United States)

    Partington, Sara L.; Kwong, Raymond Y.

    2014-01-01

    The prevalence of heart failure due to coronary artery disease continues to increase, and it portends a worse prognosis than non-ischemic cardiomyopathy. Revascularization improves prognosis in these high-risk patients who have evidence of viability; therefore, optimal assessment of myocardial viability remains essential. Multiple imaging modalities exist for differentiating viable myocardium from scar in territories with contractile dysfunction. Given the multiple modalities available, choosing the best modality for a specific patient can be a daunting task. In this review, the physiology of myocardial hibernation and stunning will be reviewed. All the current methods available for assessing viability including echocardiography, cardiac magnetic resonance imaging, nuclear imaging with single photon emission tomography and positron emission tomography imaging and cardiac computed tomography will be reviewed. The effectiveness of the various techniques will be compared, and the limitations of the current literature will be discussed. PMID:21069458

  18. Does the balanced scorecard support organizational viability?

    NARCIS (Netherlands)

    Achterbergh, J.M.I.M.; Beeres, R.J.M.; Vriens, D.J.

    2003-01-01

    In this paper we assess whether the balanced scorecard (BSC) supports the necessary functions for organizational viability. To this purpose, we use the viable system model (VSM) as a means to describe the functions required for organizational viability. Then we use the VSM as a template to assess

  19. Pollen viability and membrane lipid composition

    NARCIS (Netherlands)

    Bilsen, van D.G.J.L.

    1993-01-01

    In this thesis membrane lipid composition is studied in relation to pollen viability during storage. Chapter 1 reviews pollen viability, membranes in the dry state and membrane changes associated with cellular aging. This chapter is followed by a study of age-related changes in phospholipid

  20. Banana nectar as a medium for testing pollen viability and ...

    African Journals Online (AJOL)

    Administrator

    2007-05-16

    May 16, 2007 ... A quick and reliable method for evaluating pollen quality is essential in a breeding program, especially in a crop such as banana that is characterized by high male and female sterility. In this study the germination and viability of banana pollen was evaluated in a sucrose solution and diluted banana nectar.

  1. Banana nectar as a medium for testing pollen viability and ...

    African Journals Online (AJOL)

    A quick and reliable method for evaluating pollen quality is essential in a breeding program, especially in a crop such as banana that is characterized by high male and female sterility. In this study the germination and viability of banana pollen was evaluated in a sucrose solution and diluted banana nectar. Twenty banana ...

  2. Growth and ethanol fermentation ability on hexose and pentose sugars and glucose effect under various conditions in thermotolerant yeast Kluyveromyces marxianus

    Energy Technology Data Exchange (ETDEWEB)

    Rodrussamee, Nadchanok; Hirata, Katsushi; Suprayogi [Yamaguchi Univ., Ube (Japan). Graduate School of Medicine; Lertwattanasakul, Noppon; Kosaka, Tomoyuki [Yamaguchi Univ. (Japan). Faculty of Agriculture; Limtong, Savitree [Kasetsart Univ., Bangkok (Thailand). Faculty of Science; Yamada, Mamoru [Yamaguchi Univ., Ube (Japan). Graduate School of Medicine; Yamaguchi Univ. (Japan). Faculty of Agriculture

    2011-05-15

    Ethanol fermentation ability of the thermotolerant yeast Kluyveromyces marxianus, which is able to utilize various sugars including glucose, mannose, galactose, xylose, and arabinose, was examined under shaking and static conditions at high temperatures. The yeast was found to produce ethanol from all of these sugars except for arabinose under a shaking condition but only from hexose sugars under a static condition. Growth and sugar utilization rate under a static condition were slower than those under a shaking condition, but maximum ethanol yield was slightly higher. Even at 40 C, a level of ethanol production similar to that at 30 C was observed except for galactose under a static condition. Glucose repression on utilization of other sugars was observed, and it was more evident at elevated temperatures. Consistent results were obtained by the addition of 2-deoxyglucose. The glucose effect was further examined at a transcription level, and it was found that KmGAL1 for galactokinase and KmXYL1 for xylose reductase for galactose and xylose/arabinose utilization, respectively, were repressed by glucose at low and high temperatures, but KmHXK2 for hexokinase was not repressed. We discuss the possible mechanism of glucose repression and the potential for utilization of K. marxianus in high-temperature fermentation with mixed sugars containing glucose. (orig.)

  3. Evaluation of in vitro gas production and nutrient digestibility of complete diets supplemented with different levels of thermotolerant yeast in Nellore rams

    Directory of Open Access Journals (Sweden)

    Ch. Harikrishna

    degradable organic matter were recorded for rations D to D over control, whereas the difference among D to D diets was not 2 6 2 4 significant. Significant (P<0.05 partitioning factor and efficiency of microbial biomass synthesis (EMBP values for rations D and D and significant (P<0.01 microbial biomass production for D to D diets, recorded. Significant (P<0.01 nitrogen 2 3 2 6 (N retention was recorded in yeast supplemented rations with high EMBP than control. Dry matter intake (DMI, DMI (% b. wt., DMI/kg W0.75 and the digestibility of dry matter, organic matter, crude protein, crude fibre, ether extract (P<0.05, nitrogen free extract and fiber fractions were significant (P<0.01 on rations D to D over control, while the difference among 2 4 rations D to D was not significant. All rams were in positive N balance and was higher (P<0.01 on rations D to D over 2 4 2 4 control. The digestible crude protein (DCP and ME were higher (P<0.01 on rations D to D indicating all rams were 2 4 adequately met with DCP and ME requirements as suggested by ICAR, India. Conclusions: The present study demonstrated the potential of theromotolerant yeast at 1 g/kg to 3g/kg level in improving digestibility of nutrients, intake of DCP and ME and N retention without affecting health of rams under study. Thus, incorporation of 1 g/kg level of thermotolerant probiotic yeast in complete diet for lambs appears to be beneficial for livestock producers. [Vet. World 2012; 5(8.000: 477-485

  4. Determination of thermotolerant coliforms present in coconut water produced and bottled in the Northeast of Brazil

    Directory of Open Access Journals (Sweden)

    Vandbergue Santos Pereira

    2017-10-01

    Full Text Available Abstract Coconut water is considered to be a natural isotonic drink and its marketing is gradually increasing. The objective of the present study was to evaluate the microbiological quality of the coconut water produced and bottled in the Northeast of Brazil. Products form ten industries from different states in the Northeast of Brazil were analyzed. The most probable number (MPN method was used to quantify the coliforms. Samples showing positive for coliforms were seeded on ChromAgar Orient plates and the bacteria identified from isolated colonies using the automated system Vitek 2 (BioMérieux, according to the manufacturer's instructions for the preparation of the inoculum, incubation, reading and interpretation. The samples showed thermotolerant coliform counts between 6.0×102 and 2.6×104 MPN/100 mL. The presence of Klebsiella pneumoniae, Morganella morganii and Providencia alcalifaciens was observed. The implementation of preventive methods and monitoring of the water quality by the industries is required.

  5. Isolation, Production, and Characterization of Thermotolerant Xylanase from Solvent Tolerant Bacillus vallismortis RSPP-15

    Directory of Open Access Journals (Sweden)

    Rajeeva Gaur

    2015-01-01

    Full Text Available Sixty bacterial strains isolated from the soils sample in the presence of organic solvent were screened for xylanase production. Among them, strain RSPP-15 showed the highest xylanase activity which was identified as Bacillus vallismortis. The isolate showed maximum xylanase production (3768 U/mL in the presence of birch wood xylan and beef extract at 55°C pH 7.0 within 48 h of incubation. The enzyme activity and stability were increased 181.5, 153.7, 147.2, 133.6, and 127.9% and 138.2, 119.3, 113.9, 109, and 104.5% in the presence of Co2+, Ca2+, Mg+2, Zn+2, and Fe+3 ions (10 mM. Xylanase activity and stability were strongly inhibited in the presence of Hg and Cu ions. The enzyme was also stable in the presence of 30% of n-dodecane, isooctane, n-decane, xylene, toluene, n-hexane, n-butanol, and cyclohexane, respectively. The presence of benzene, methanol, and ethanol marginally reduced the xylanase stability, respectively. This isolate may be useful in several industrial applications owing to its thermotolerant and organic solvent resistance characteristics.

  6. Can E. coli or thermotolerant coliform concentrations predict pathogen presence or prevalence in irrigation waters?

    Science.gov (United States)

    Pachepsky, Yakov; Shelton, Daniel; Dorner, Sarah; Whelan, Gene

    2016-05-01

    An increase in food-borne illnesses in the United States has been associated with fresh produce consumption. Irrigation water presents recognized risks for microbial contamination of produce. Water quality criteria rely on indicator bacteria. The objective of this review was to collate and summarize experimental data on the relationships between pathogens and thermotolerant coliform (THT) and/or generic E. coli, specifically focusing on surface fresh waters used in or potentially suitable for irrigation agriculture. We analyzed peer-reviewed publications in which concentrations of E. coli or THT coliforms in surface fresh waters were measured along with concentrations of one or more of waterborne and food-borne pathogenic organisms. The proposed relationships were significant in 35% of all instances and not significant in 65% of instances. Coliform indicators alone cannot provide conclusive, non-site-specific and non-pathogen-specific information about the presence and/or concentrations of most important pathogens in surface waters suitable for irrigation. Standards of microbial water quality for irrigation can rely not only on concentrations of indicators and/or pathogens, but must include references to crop management. Critical information on microbial composition of actual irrigation waters to support criteria of microbiological quality of irrigation waters appears to be lacking and needs to be collected.

  7. Prevalence of thermotolerant Campylobacter in broiler flocks and broiler carcasses in Italy

    Directory of Open Access Journals (Sweden)

    Giacomo Migliorati

    2010-12-01

    Full Text Available In accordance with European Union regulations, from 5 February until 15 December 2008, sampling and analysis activities were conducted in Italy to assess the extent of contamination caused by thermotolerant Campylobacter in broiler chickens farmed nationwide. The survey involved 48 poultry slaughterhouses distributed across eleven regions of Italy, where the caeca and carcasses of 393 slaughter batches were sampled. A total of 284 batches (72.3% gave positive results for Campylobacter spp. as follows: 52.1% were contaminated by C. jejuni, 55.6% by C. coli and 1.1% by C. lari. C. jejuni and C. coli were isolated together in 37 batches (13% of positive results. Campylobacter spp. was isolated only from the caeca in 251 slaughter batches (63.9% including caecal isolates of C. jejuni (48.2%, C. coli (50.6%, and C. lari (1.2%. Carcasses from 182 batches (46.3% were contaminated by C. jejuni in 40.7% of cases, C. coli in 57.7% and the absence of C. lari from all batches examined. The contamination level observed in the carcasses ranged between 10 and 1.6 × 107 cfu/g.

  8. Bioconversion of Airborne Methylamine by Immobilized Recombinant Amine Oxidase from the Thermotolerant Yeast Hansenula polymorpha

    Directory of Open Access Journals (Sweden)

    Sasi Sigawi

    2014-01-01

    Full Text Available Aliphatic amines, including methylamine, are air-pollutants, due to their intensive use in industry and the natural degradation of proteins, amino acids, and other nitrogen-containing compounds in biological samples. It is necessary to develop systems for removal of methylamine from the air, since airborne methylamine has a negative effect on human health. The primary amine oxidase (primary amine : oxygen oxidoreductase (deaminating or amine oxidase, AMO; EC 1.4.3.21, a copper-containing enzyme from the thermotolerant yeast Hansenula polymorpha which was overexpressed in baker’s yeast Saccharomyces cerevisiae, was tested for its ability to oxidize airborne methylamine. A continuous fluidized bed bioreactor (CFBR was designed to enable bioconversion of airborne methylamine by AMO immobilized in calcium alginate (CA beads. The results demonstrated that the bioreactor with immobilized AMO eliminates nearly 97% of the airborne methylamine. However, the enzymatic activity of AMO causes formation of formaldehyde. A two-step bioconversion process was therefore proposed. In the first step, airborne methylamine was fed into a CFBR which contained immobilized AMO. In the second step, the gas flow was passed through another CFBR, with alcohol oxidase from the yeast H. polymorpha immobilized in CA, in order to decompose the formaldehyde formed in the first step. The proposed system provided almost total elimination of the airborne methylamine and the formaldehyde.

  9. Recovery of heat shock-triggered released apoplastic Ca2+ accompanied by pectin methylesterase activity is required for thermotolerance in soybean seedlings.

    Science.gov (United States)

    Wu, Hui-Chen; Hsu, Shih-Feng; Luo, Dan-Li; Chen, Shiang-Jiuun; Huang, Wen-Dar; Lur, Huu-Sheng; Jinn, Tsung-Luo

    2010-06-01

    Synthesis of heat shock proteins (HSPs) in response to heat shock (HS) is essential for thermotolerance. The effect of a Ca(2+) chelator, EGTA, was investigated before a lethal HS treatment in soybean (Glycine max) seedlings with acquired thermotolerance induced by preheating. Such seedlings became non-thermotolerant with EGTA treatment. The addition of Ca(2+), Sr(2+) or Ba(2+) to the EGTA-treated samples rescued the seedlings from death by preventing the increased cellular leakage of electrolytes, amino acids, and sugars caused by EGTA. It was confirmed that EGTA did not affect HSP accumulation and physiological functions but interfered with the recovery of HS-released Ca(2+) concentration which was required for thermotolerance. Pectin methylesterase (PME, EC 3.1.1.11), a cell wall remodelling enzyme, was activated in response to HS, and its elevated activity caused an increased level of demethylesterified pectin which was related to the recovery of the HS-released Ca(2+) concentration. Thus, the recovery of HS-released Ca(2+) in Ca(2+)-pectate reconstitution through PME activity is required for cell wall remodelling during HS in soybean which, in turn, retains plasma membrane integrity and co-ordinates with HSPs to confer thermotolerance.

  10. Population-specific life histories contribute to metapopulation viability

    Science.gov (United States)

    Halsey, Samniqueka J.; Bell, Timothy J.; McEachern, A. Kathryn; Pavlovic, Noel B.

    2016-01-01

    Restoration efforts can be improved by understanding how variations in life-history traits occur within populations of the same species living in different environments. This can be done by first understanding the demographic responses of natural occurring populations. Population viability analysis continues to be useful to species management and conservation with sensitivity analysis aiding in the understanding of population dynamics. In this study, using life-table response experiments and elasticity analyses, we investigated how population-specific life-history demographic responses contributed to the metapopulation viability of the Federally threatened Pitcher's thistle (Cirsium pitcheri). Specifically, we tested the following hypotheses: (1) Subpopulations occupying different environments within a metapopulation have independent demographic responses and (2) advancing succession results in a shift from a demographic response focused on growth and fecundity to one dominated by stasis. Our results showed that reintroductions had a positive contribution to the metapopulation growth rate as compared to native populations which had a negative contribution. We found no difference in succession on the contribution to metapopulation viability. In addition, we identified distinct population-specific contributions to metapopulation viability and were able to associate specific life-history demographic responses. For example, the positive impact of Miller High Dunes population on the metapopulation growth rate resulted from high growth contributions, whereas increased time of plant in stasis for the State Park Big Blowout population resulted in negative contributions. A greater understanding of how separate populations respond in their corresponding environment may ultimately lead to more effective management strategies aimed at reducing extinction risk. We propose the continued use of sensitivity analyses to evaluate population-specific demographic influences on

  11. Enhanced production of raw starch degrading enzyme using agro-industrial waste mixtures by thermotolerant Rhizopus microsporus for raw cassava chip saccharification in ethanol production.

    Science.gov (United States)

    Trakarnpaiboon, Srisakul; Srisuk, Nantana; Piyachomkwan, Kuakoon; Sakai, Kenji; Kitpreechavanich, Vichien

    2017-09-14

    In the present study, solid-state fermentation for the production of raw starch degrading enzyme was investigated by thermotolerant Rhizopus microsporus TISTR 3531 using a combination of agro-industrial wastes as substrates. The obtained crude enzyme was applied for hydrolysis of raw cassava starch and chips at low temperature and subjected to nonsterile ethanol production using raw cassava chips. The agro-industrial waste ratio was optimized using a simplex axial mixture design. The results showed that the substrate mixture consisting of rice bran:corncob:cassava bagasse at 8 g:10 g:2 g yielded the highest enzyme production of 201.6 U/g dry solid. The optimized condition for solid-state fermentation was found as 65% initial moisture content, 35°C, initial pH of 6.0, and 5 × 10 6 spores/mL inoculum, which gave the highest enzyme activity of 389.5 U/g dry solid. The enzyme showed high efficiency on saccharification of raw cassava starch and chips with synergistic activities of commercial α-amylase at 50°C, which promotes low-temperature bioethanol production. A high ethanol concentration of 102.2 g/L with 78% fermentation efficiency was achieved from modified simultaneous saccharification and fermentation using cofermentation of the enzymatic hydrolysate of 300 g raw cassava chips/L with cane molasses.

  12. Terminology for pregnancy loss prior to viability

    DEFF Research Database (Denmark)

    Kolte, A M; Bernardi, L A; Christiansen, O B

    2015-01-01

    Pregnancy loss prior to viability is common and research in the field is extensive. Unfortunately, terminology in the literature is inconsistent. The lack of consensus regarding nomenclature and classification of pregnancy loss prior to viability makes it difficult to compare study results from...... different centres. In our opinion, terminology and definitions should be based on clinical findings, and when possible, transvaginal ultrasound. With this Early Pregnancy Consensus Statement, it is our goal to provide clear and consistent terminology for pregnancy loss prior to viability....

  13. Morphology and viability of castor bean genotypes pollen grains

    Directory of Open Access Journals (Sweden)

    Maria Selma Alves Silva Diamantino

    2016-01-01

    Full Text Available The objective of this work was to characterize the morphology and viability of the pollen of 15 genotypes of castor bean (Ricinus communis L. and to generate information that can assist in the selection of highly promising male parents for future use in genetic improvement programs aimed at producing seeds for oil extraction. Acetolysis and scanning electron microscopy was used to characterize the morphology of the pollen. The viability of the pollen grains was estimated by in vitro germination and colorimetric analysis (acetocarmine 2% and 2, 3, 5-triphenyltetrazolium chloride 1%. For the in vitro germination, pollen grains were grown in 10 types of solidified culture medium consisting of different concentrations of sucrose, boric acid, calcium nitrate, magnesium sulfate and potassium nitrate. The pollen grains had the following characteristics: medium size, isopolar and subspheroidal shape, radial symmetry, circular ambit, 3-colporate, elongated endoapertures, tectate exine and granulated sexine. The acetocarmine dye overestimated pollen viability. The media M5 and M8 were the most efficient at promoting the germination of pollen grains. The studied genotypes had high levels of viability and can therefore be used as male parents in genetic improvement programs.

  14. A comparison of assays measuring the viability of Legionella ...

    Science.gov (United States)

    Background: The relatively high prevalence of Legionella pneumophila in premise plumbing systems has been widely reported. Published reports indicate Legionella has a comparatively high resistance to chlorine and moreover has the ability to grow in phagocytic amoeba which could provide additional protection in chlorinated drinking water distribution systems. Copper-Silver (Cu-Ag) ionization treatment systems are commercially available for use in large building water systems to help control the risks from Legionella bacteria. The objectives of this study were to develop and optimize Legionella viability assays and use them to investigate the viability of Legionella bacteria after exposure to water treated with coppper and silver ions. Methods: Log phase L. pneumophila cells were used in all experiments and were generated by incubation at 35C for 48 hours in buffered yeast extract broth. Viability assays used included plating on buffered charcoal yeast extract agar to determine the number of culturable cells and treating cells with propidium monoazide (PMA) or ethidium monoazide (EMA) followed by quantitative PCR targeting mip gene of L. pneumophila. The qPCR viability assays were optimized using L. pneumophila inactivated by heat treatment at 65C for 60 min. The effectiveness of Cu-Ag ionization treatment was studied by inoculating L. pneumonia at 105 CFU/mL in water collected directly from a building water system that employed this technology and incubat

  15. A collaborative study on a Nordic standard protocol for detection and enumeration of thermotolerant Campylobacter in food (NMKL 119, 3. Ed., 2007)

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Bengtsson, Anja; Hansen, Tina Beck

    2007-01-01

    A Nordic standard protocol for detection and enumeration of thermotolerant Campylobacter in food has been elaborated (NMKL 119, 3. Ed., 2007). Performance and precision characteristics of this protocol were evaluated in a collaborative study with participation of 14 laboratories from seven European.......4-86.6%), respectively. The sensitivity was not influenced by food matrix (P=0.359), but was significantly lower for C. coli compared to C. jejuni (P= 0.007) and for concentrations below 1.4 cfu g(-1) (P... variation between laboratories can be explained by the general difficulties in handling Campylobacter In conclusion, NMKL 119, 3. Ed., 2007, is regarded as an acceptable protocol for detection of thermotolerant Campylobacter at concentrations above 25 cfu g(-1) and also for enumeration of thermotolerant...

  16. Near viability for fully nonlinear differential inclusions

    National Research Council Canada - National Science Library

    Irina Căpraru; Alina Lazu

    2014-01-01

    .... We establish a viability result under Lipschitz hypothesis on F, that consists in proving the existence of solutions of the differential inclusion above, starting from a given set, which remain...

  17. Poxvirus viability and signatures in historical relics

    National Research Council Canada - National Science Library

    McCollum, Andrea M; Li, Yu; Wilkins, Kimberly; Karem, Kevin L; Davidson, Whitni B; Paddock, Christopher D; Reynolds, Mary G; Damon, Inger K

    2014-01-01

    Although it has been >30 years since the eradication of smallpox, the unearthing of well-preserved tissue material in which the virus may reside has called into question the viability of variola virus decades or centuries...

  18. Intraspecific variation in pollen viability, germination and ...

    African Journals Online (AJOL)

    Oleaceae) cultivars 'Koroneiki', 'Mastoidis' and 'Kalamata' was studied with scanning electron microscopy to identify genotype- distinguishing characters that could be employed for morphological cultivar discrimination. Pollen viability and germination ...

  19. A new search for thermotolerant yeasts, its characterization and optimization using response surface methodology for ethanol production.

    Science.gov (United States)

    Arora, Richa; Behera, Shuvashish; Sharma, Nilesh K; Kumar, Sachin

    2015-01-01

    The progressive rise in energy crisis followed by green house gas (GHG) emissions is serving as the driving force for bioethanol production from renewable resources. Current bioethanol research focuses on lignocellulosic feedstocks as these are abundantly available, renewable, sustainable and exhibit no competition between the crops for food and fuel. However, the technologies in use have some drawbacks including incapability of pentose fermentation, reduced tolerance to products formed, costly processes, etc. Therefore, the present study was carried out with the objective of isolating hexose and pentose fermenting thermophilic/thermotolerant ethanologens with acceptable product yield. Two thermotolerant isolates, NIRE-K1 and NIRE-K3 were screened for fermenting both glucose and xylose and identified as Kluyveromyces marxianus NIRE-K1 and K. marxianus NIRE-K3. After optimization using Face-centered Central Composite Design (FCCD), the growth parameters like temperature and pH were found to be 45.17°C and 5.49, respectively for K. marxianus NIRE-K1 and 45.41°C and 5.24, respectively for K. marxianus NIRE-K3. Further, batch fermentations were carried out under optimized conditions, where K. marxianus NIRE-K3 was found to be superior over K. marxianus NIRE-K1. Ethanol yield (Y x∕s ), sugar to ethanol conversion rate (%), microbial biomass concentration (X) and volumetric product productivity (Q p ) obtained by K. marxianus NIRE-K3 were found to be 9.3, 9.55, 14.63, and 31.94% higher than that of K. marxianus NIRE-K1, respectively. This study revealed the promising potential of both the screened thermotolerant isolates for bioethanol production.

  20. A new search for thermotolerant yeasts, its characterization and optimization using response surface methodology for ethanol production

    Directory of Open Access Journals (Sweden)

    Richa eArora

    2015-09-01

    Full Text Available The progressive rise in energy crisis followed by green house gas (GHG emissions is serving as the driving force for bioethanol production from renewable resources. Current bioethanol research focuses on lignocellulosic feedstocks as these are abundantly available, renewable, sustainable and exhibit no competition between the crops for food and fuel. However, the technologies in use have some drawbacks including incapability of pentose fermentation, reduced tolerance to products formed, costly processes, etc. Therefore, the present study was carried out with the objective of isolating hexose and pentose fermenting thermophilic/ thermotolerant ethanologens with acceptable product yield. Two thermotolerant isolates, NIRE-K1 and NIRE-K3 were screened for fermenting both glucose and xylose and identified as Kluyveromyces marxianus NIRE-K1 and K. marxianus NIRE-K3. After optimization using FCCD (Face-centered Central Composite Design, the growth parameters like temperature and pH were found to be 45.17 oC and 5.49, respectively for K. marxianus NIRE-K1 and 45.41 oC and 5.24, respectively for K. marxianus NIRE-K3. Further, batch fermentations were carried out under optimized conditions, where K. marxianus NIRE-K3 was found to be superior over K. marxianus NIRE-K1. Ethanol yield (Yx/s, sugar to ethanol conversion rate (%, microbial biomass concentration (X and volumetric product productivity (Qp obtained by K. marxianus NIRE-K3 were found to be 9.3%, 9.55%, 14.63% and 31.94% higher than that of K. marxianus NIRE-K1, respectively. This study revealed the promising potential of both the screened thermotolerant isolates for bioethanol production.

  1. Hyperthermia combined with 5-fluorouracil promoted apoptosis and enhanced thermotolerance in human gastric cancer cell line SGC-7901

    Directory of Open Access Journals (Sweden)

    Liu T

    2015-05-01

    Full Text Available Tao Liu,* Yan-Wei Ye,* A-li Zhu, Zhen Yang, Yang Fu, Chong-Qing Wei, Qi Liu, Chun-Lin Zhao, Guo-Jun Wang, Xie-Fu Zhang Department of Gastrointestinal Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, People’s Republic of China *These authors contributed equally to this work Abstract: This study was designed to investigate the proliferation inhibition and apo­ptosis-promoting effect under hyperthermia and chemotherapy treatment, at cellular level. Human gastric cancer cell line SGC-7901 was cultivated with 5-fluorouracil at different temperatures. Cell proliferation and apoptosis were determined, and expression of Bcl-2 and HSP70 was measured at different treatments. Cell survival rates and inhibition rates in chemotherapy group, thermotherapy group, and thermo-chemotherapy group were drastically lower than the control group (P<0.05. For tumor cells in the thermo-chemotherapy group, survival rates and inhibition rates at three different temperatures were all significantly lower than those in chemotherapy group and thermotherapy group (P<0.05. 5-Fluorouracil induced apoptosis of SGC-7901 cells with a strong temperature dependence, which increased gradually with increase in temperature. At 37°C and 43°C there were significant differences between the thermotherapy group and chemotherapy group and between the thermo-chemotherapy group and thermotherapy group (P<0.01. The expression of Bcl-2 was downregulated and HSP70 was upregulated, with increase in temperature in all groups. Cell apoptosis was not significant at 46°C (P>0.05, which was probably due to thermotolerance caused by HSP70 accumulation. These results suggested that hyperthermia combined with 5-fluorouracil had a synergistic effect in promoting apoptosis and enhancing thermotolerance in gastric cancer cell line SGC-7901. Keywords: gastric cancer, thermotherapy, 5-fluorouracil, Bcl-2, HSP70, thermotolerance

  2. Probiotic viability – does it matter?

    OpenAIRE

    Sampo J. Lahtinen

    2012-01-01

    Probiotics are viable by definition, and viability of probiotics is often considered to be a prerequisite for the health benefits. Indeed, the overwhelming majority of clinical studies in the field have been performed with viable probiotics. However, it has also been speculated that some of the mechanisms behind the probiotic health effects may not be dependent on the viability of the cells, and therefore is also possible that also nonviable probiotics could have some health benefits. The eff...

  3. Isolation and Characterization of a Thermotolerant Ammonia-Oxidizing Bacterium Nitrosomonas sp. JPCCT2 from a Thermal Power Station

    OpenAIRE

    Itoh, Yoshikane; Sakagami, Keiko; Uchino, Yoshihito; Boonmak, Chanita; Oriyama, Tetsuro; Tojo, Fuyumi; Matsumoto, Mitsufumi; Morikawa, Masaaki

    2013-01-01

    A thermotolerant ammonia-oxidizing bacterium strain JPCCT2 was isolated from activated sludge in a thermal power station. Cells of JPCCT2 are short non-motile rods or ellipsoidal. Molecular phylogenetic analysis of 16S rRNA gene sequences demonstrated that JPCCT2 belongs to the genus Nitrosomonas with the highest similarity to Nitrosomonas nitrosa Nm90 (100%), Nitrosomonas sp. Nm148 (99.7%), and Nitrosomonas communis Nm2 (97.7%). However, G+C content of JPCCT2 DNA was 49.1 mol% and clearly di...

  4. Low-dose adenosine stress echocardiography: Detection of myocardial viability

    Directory of Open Access Journals (Sweden)

    Nedeljkovic Milan

    2003-06-01

    Full Text Available Abstract Objective The aim of this study was to evaluate the diagnostic potential of low-dose adenosine stress echocardiography in detection of myocardial viability. Background Vasodilation through low dose dipyridamole infusion may recruit contractile reserve by increasing coronary flow or by increasing levels of endogenous adenosine. Methods Forty-three patients with resting dyssynergy, due to previous myocardial infarction, underwent low-dose adenosine (80, 100, 110 mcg/kg/min in 3 minutes intervals echocardiography test. Gold standard for myocardial viability was improvement in systolic thickening of dyssinergic segments of ≥ 1 grade at follow-up. Coronary angiography was done in 41 pts. Twenty-seven patients were revascularized and 16 were medically treated. Echocardiographic follow up data (12 ± 2 months were available in 24 revascularized patients. Results Wall motion score index improved from rest 1.55 ± 0.30 to 1.33 ± 0.26 at low-dose adenosine (p Conclusion Low-dose adenosine stress echocardiography test has high diagnostic potential for detection of myocardial viability in the group of patients with left ventricle dysfunction due to previous myocardial infarction. Low dose adenosine stress echocardiography may be adequate alternative to low-dose dobutamine test for evaluation of myocardial viability.

  5. Effect of salt hyperosmotic stress on yeast cell viability

    Directory of Open Access Journals (Sweden)

    Logothetis Stelios

    2007-01-01

    Full Text Available During fermentation for ethanol production, yeasts are subjected to different kinds of physico-chemical stresses such as: initially high sugar concentration and low temperature; and later, increased ethanol concentrations. Such conditions trigger a series of biological responses in an effort to maintain cell cycle progress and yeast cell viability. Regarding osmostress, many studies have been focused on transcriptional activation and gene expression in laboratory strains of Saccharomyces cerevisiae. The overall aim of this present work was to further our understanding of wine yeast performance during fermentations under osmotic stress conditions. Specifically, the research work focused on the evaluation of NaCl-induced stress responses of an industrial wine yeast strain S. cerevisiae (VIN 13, particularly with regard to yeast cell growth and viability. The hypothesis was that osmostress conditions energized specific genes to enable yeast cells to survive under stressful conditions. Experiments were designed by pretreating cells with different sodium chloride concentrations (NaCl: 4%, 6% and 10% w/v growing in defined media containing D-glucose and evaluating the impact of this on yeast growth and viability. Subsequent fermentation cycles took place with increasing concentrations of D-glucose (20%, 30%, 40% w/v using salt-adapted cells as inocula. We present evidence that osmostress induced by mild salt pre-treatments resulted in beneficial influences on both cell viability and fermentation performance of an industrial wine yeast strain.

  6. Association of ATP1A1 gene polymorphism with thermotolerance in Tharparkar and Vrindavani cattle

    Directory of Open Access Journals (Sweden)

    Neeraj Kashyap

    2015-07-01

    Full Text Available Aim: One of the major biochemical aspects of thermoregulation is equilibrium of ion gradient across biological membranes. Na+/K+-ATPase, a member of P type-ATPase family, is a major contributor to the mechanism that actively controls crossmembrane ion gradient. Thus, we examined ATP1A1 gene that encodes alpha-1 chain of Na+/K+-ATPase, for genetic polymorphisms. Materials and Methods: A total of 100 Vrindavani (composite cross strain of Hariana x Holstein-Friesian/Brown Swiss/Jersey and 64 Tharparkar (indigenous cattle were screened for genetic polymorphism in ATP1A1 gene, using polymerase chain reaction single-strand conformation polymorphism and DNA sequencing. For association studies, rectal temperature (RT and respiration rate (RR of all animals were recorded twice daily for 3 seasons. Results: A SNP (C2789A was identified in exon 17 of ATP1A1 gene. Three genotypes namely CC, CA, and AA were observed in both, Vrindavani and Tharparkar cattle. The gene frequencies in Tharparkar and Vrindavani for allele A were 0.51 and 0.48, and for allele C were 0.49 and 0.52, respectively, which remained at intermediate range. Association study of genotypes with RT and RR in both cattle population revealed that the animals with genotype CC exhibited significantly lower RT and higher heat tolerance coefficient than CA and AA genotypes. Conclusion: Differential thermoregulation between different genotypes of ATP1A1 gene indicate that the ATP1A1 gene could be potentially contributing to thermotolerance in both, Tharparkar, an indigenous breed and Vrindavani, a composite crossbred cattle.

  7. Biosorption of heavy metal by thermotolerant polymerproducing bacterial cells and the bioflocculant

    Directory of Open Access Journals (Sweden)

    Saithong Kaewchai

    2002-07-01

    Full Text Available Three strains of thermotolerant polymer-producing bacteria; Bacillus subtilis WD 90, Bacillus subtilis SM 29, and Enterobacter agglomerans SM 38 as well as their biofloculants were used to investigate on the adsorption of heavy metal, nickel and cadmium. The effects of pH and concentrations of heavy metal were investigated. The optimum pH for nickel and cadmium adsorption by the dried cells of E. agglomerans SM 38 were found to be 7.0 (25.5% removal and 8.0 (32% removal, respectively. For B. subtilis WD 90 and B. subtilis SM 29, the optimum pH at 8.0 exhibited the nickel removal of 27% and 25%, respectively, and cadmium removal of 28% and 28.5%, respectively. The heavy metal adsorption by the dried cells and wet cells of E. agglomerans SM 38 were slightly increased with increasing initial concentrations of nickel and cadmium up to 60 and 30 ppm, respectively. The bioflocculant of B. subtilis WD 90 and B. subtilis SM 29 showed the highest nickel removal of 90.7% and 87.0% respectively, while the cadmium removal was 90.9 and 91.4%, respectively. The optimum pH for adsorption of both nickel and cadmium by the bioflocculant of E. agglomerans SM 38 was 7.0 with the removal of 92.8 and 84.2%, respectively. The optimum nickel concentration for adsorption by the bioflocculant of E. agglomerans SM 38 was 10 ppm, with the removal of 92.5%, and rather stable up to 60 ppm. The optimum cadmium concentration for adsorption by the bioflocculant of B. subtilis SM 29 was 60 ppm at pH 8.0 with the removal of 85.7%. Therefore, the bioflocculant of the three isolates gave higher heavy metal adsorption than the cells.

  8. Characterization of alcohol dehydrogenase 3 of the thermotolerant methylotrophic yeast Hansenula polymorpha.

    Science.gov (United States)

    Suwannarangsee, Surisa; Kim, Seonghun; Kim, Oh-Cheol; Oh, Doo-Byoung; Seo, Jeong-Woo; Kim, Chul Ho; Rhee, Sang Ki; Kang, Hyun Ah; Chulalaksananukul, Warawut; Kwon, Ohsuk

    2012-11-01

    In this study, we identified and characterized mitochondrial alcohol dehydrogenase 3 from the thermotolerant methylotrophic yeast Hansenula polymorpha (HpADH3). The amino acid sequence of HpADH3 shares over 70% of its identity with the alcohol dehydrogenases of other yeasts and exhibits the highest similarity of 91% with the alcohol dehydrogenase 1 of H. polymorpha. However, unlike the cytosolic HpADH1, HpADH3 appears to be a mitochondrial enzyme, as a mitochondrial targeting extension exists at its N terminus. The recombinant HpADH3 overexpressed in Escherichia coli showed similar catalytic efficiencies for ethanol oxidation and acetaldehyde reduction. The HpADH3 displayed substrate specificities with clear preferences for medium chain length primary alcohols and acetaldehyde for an oxidation reaction and a reduction reaction, respectively. Although the H. polymorpha ADH3 gene was induced by ethanol in the culture medium, both an ADH isozyme pattern analysis and an ADH activity assay indicated that HpADH3 is not the major ADH in H. polymorpha DL-1. Moreover, HpADH3 deletion did not affect the cell growth on different carbon sources. However, when the HpADH3 mutant was complemented by an HpADH3 expression cassette fused to a strong constitutive promoter, the resulting strain produced a significantly increased amount of ethanol compared to the wild-type strain in a glucose medium. In contrast, in a xylose medium, the ethanol production was dramatically reduced in an HpADH3 overproduction strain compared to that in the wild-type strain. Taken together, our results suggest that the expression of HpADH3 would be an ideal engineering target to develop H. polymorpha as a substrate specific bioethanol production strain.

  9. High-Temperature-Induced Defects in Tomato (Solanum lycopersicum) Anther and Pollen Development Are Associated with Reduced Expression of B-Class Floral Patterning Genes

    Science.gov (United States)

    Kristensen, Lieke; Wolters-Arts, Mieke; de Groot, Peter F. M.; Jansma, Stuart Y.; Mariani, Celestina; Park, Sunghun

    2016-01-01

    Sexual reproduction is a critical process in the life-cycle of plants and very sensitive to environmental perturbations. To better understand the effect of high temperature on plant reproduction, we cultivated tomato (Solanum lycopersicum) plants in continuous mild heat. Under this condition we observed a simultaneous reduction in pollen viability and appearance of anthers with pistil-like structures, while in a more thermotolerant genotype, both traits were improved. Ectopic expression of two pistil-specific genes, TRANSMITTING TISSUE SPECIFIC and TOMATO AGAMOUS LIKE11, in the anthers confirmed that the anthers had gained partial pistil identity. Concomitantly, expression of the B-class genes TOMATO APETALA3, TOMATO MADS BOX GENE6 (TM6) and LePISTILLATA was reduced in anthers under continuous mild heat. Plants in which TM6 was partially silenced reacted hypersensitively to temperature elevation with regard to the frequency of pistilloid anthers, pollen viability and pollen quantity. Taken together, these results suggest that high-temperature-induced down-regulation of tomato B-class genes contributes to anther deformations and reduced male fertility. Improving our understanding of how temperature perturbs the molecular mechanisms of anther and pollen development will be important in the view of maintaining agricultural output under current climate changes. PMID:27936079

  10. Azetidine-induced accumulation of class I small heat shock proteins in the soluble fraction provides thermotolerance in soybean seedlings.

    Science.gov (United States)

    Jinn, Tsung-Luo; Chiu, Chi-Chou; Song, Wei-Wen; Chen, Yih-Ming; Lin, Chu-Yung

    2004-12-01

    Accumulation of class I small heat shock proteins (sHSPs) is induced by the proline analog, azetidine-2-carboxylic acid (Aze) in soybean seedlings to a level similar to that induced by exposure to 40 degrees C. However, only the treatment with 10 mM Aze for 6 h and subsequently with 10 mM proline for 24 h protected the seedlings from damage during subsequent exposure to 45 degrees C as assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. A chaperone activity assay showed that the purified class I sHSPs induced by Aze were functional in vitro and protected proteins from thermal denaturation. Amino acid composition analysis indicated that Aze was not incorporated into de novo synthesized class I sHSPs. Accumulation of class I sHSPs in the soluble post-ribosomal supernatant fraction was found to be important for acquisition of thermotolerance. We suggest that both the accumulation of class I sHSPs and their presence in the soluble fraction are important for establishment of thermotolerance.

  11. Removal of total coliforms, thermotolerant coliforms, and helminth eggs in Swine production wastewater treated in anaerobic and aerobic reactors.

    Science.gov (United States)

    Zacarias Sylvestre, Silvia Helena; Lux Hoppe, Estevam Guilherme; de Oliveira, Roberto Alves

    2014-01-01

    The present work evaluated the performance of two treatment systems in reducing indicators of biological contamination in swine production wastewater. System I consisted of two upflow anaerobic sludge blanket (UASB) reactors, with 510 and 209 L in volume, being serially arranged. System II consisted of a UASB reactor, anaerobic filter, trickling filter, and decanter, being also organized in series, with volumes of 300, 190, 250, and 150 L, respectively. Hydraulic retention times (HRT) applied in the first UASB reactors were 40, 30, 20, and 11 h in systems I and II. The average removal efficiencies of total and thermotolerant coliforms in system I were 92.92% to 99.50% and 94.29% to 99.56%, respectively, and increased in system II to 99.45% to 99.91% and 99.52% to 99.93%, respectively. Average removal rates of helminth eggs in system I were 96.44% to 99.11%, reaching 100% as in system II. In reactor sludge, the counts of total and thermotolerant coliforms ranged between 10(5) and 10(9) MPN (100 mL)(-1), while helminth eggs ranged from 0.86 to 9.27 eggs g(-1) TS.

  12. Removal of Total Coliforms, Thermotolerant Coliforms, and Helminth Eggs in Swine Production Wastewater Treated in Anaerobic and Aerobic Reactors

    Science.gov (United States)

    Zacarias Sylvestre, Silvia Helena; Lux Hoppe, Estevam Guilherme; de Oliveira, Roberto Alves

    2014-01-01

    The present work evaluated the performance of two treatment systems in reducing indicators of biological contamination in swine production wastewater. System I consisted of two upflow anaerobic sludge blanket (UASB) reactors, with 510 and 209 L in volume, being serially arranged. System II consisted of a UASB reactor, anaerobic filter, trickling filter, and decanter, being also organized in series, with volumes of 300, 190, 250, and 150 L, respectively. Hydraulic retention times (HRT) applied in the first UASB reactors were 40, 30, 20, and 11 h in systems I and II. The average removal efficiencies of total and thermotolerant coliforms in system I were 92.92% to 99.50% and 94.29% to 99.56%, respectively, and increased in system II to 99.45% to 99.91% and 99.52% to 99.93%, respectively. Average removal rates of helminth eggs in system I were 96.44% to 99.11%, reaching 100% as in system II. In reactor sludge, the counts of total and thermotolerant coliforms ranged between 105 and 109 MPN (100 mL)−1, while helminth eggs ranged from 0.86 to 9.27 eggs g−1 TS. PMID:24812560

  13. LIVE/DEAD YEAST VIABILITY STAINING AS A TOOL FOR IMPROVING ARTISANAL PILSNER BEER PRODUCTION

    Directory of Open Access Journals (Sweden)

    Benedetta Bottari

    2014-10-01

    Full Text Available The production of an artisanal beer, made by brewers using traditional practices on a small scale, is founded on the empirical adjustment of parameters, including yeasts handling and serial repitching. The aim of this study was to monitor yeast viability during different stages of artisanal beer productions through the Live/Dead Yeast viability staining and to correlate it with fermentation dynamics in order to increase process standardization and to maintain the quality of final products. Yeast viability and fermentation activities were evaluated during seven fermentation cycles of an artisanal pilsner beer. Yeast inoculated with higher viability performed generally better in fermentation, resulting in faster sugar consumption, faster ethanol production and stability. Handling yeast and serial repitching based on Live/Dead viability measurements, could be the key way to ensure reliable manufacture of high quality beer and to improve process standardization particularly for microbreweries, where variability of production can be a challenging point.

  14. Viability analysis in biological evaluations: Concepts of population viability analysis, biological population, and ecological scale

    Science.gov (United States)

    Gregory D. Hayward; John R. Squires

    1994-01-01

    Environmental protection strategies often rely on environmental impact assessments. As part of the assessment process biologists are routinely asked to evaluate the effects of management actions on plants and animals. This evaluation often requires that biologists make judgments about the viability of affected populations. However, population viability...

  15. Effect of air drying on bacterial viability: A multiparameter viability assessment

    NARCIS (Netherlands)

    Nocker, A.; Fernández, P.S.; Montijn, R.; Schuren, F.

    2012-01-01

    The effect of desiccation on the viability of microorganisms is a question of great interest for a variety of public health questions and industrial applications. Although viability is traditionally assessed by plate counts, cultivation-independent methods are increasingly applied with the aim to

  16. Viability and Resilience of Languages in Competition

    Science.gov (United States)

    Chapel, Laetitia; Castelló, Xavier; Bernard, Claire; Deffuant, Guillaume; Eguíluz, Víctor M.; Martin, Sophie; Miguel, Maxi San

    2010-01-01

    We study the viability and resilience of languages, using a simple dynamical model of two languages in competition. Assuming that public action can modify the prestige of a language in order to avoid language extinction, we analyze two cases: (i) the prestige can only take two values, (ii) it can take any value but its change at each time step is bounded. In both cases, we determine the viability kernel, that is, the set of states for which there exists an action policy maintaining the coexistence of the two languages, and we define such policies. We also study the resilience of the languages and identify configurations from where the system can return to the viability kernel (finite resilience), or where one of the languages is lead to disappear (zero resilience). Within our current framework, the maintenance of a bilingual society is shown to be possible by introducing the prestige of a language as a control variable. PMID:20126655

  17. Probiotic viability – does it matter?

    Directory of Open Access Journals (Sweden)

    Sampo J. Lahtinen

    2012-06-01

    Full Text Available Probiotics are viable by definition, and viability of probiotics is often considered to be a prerequisite for the health benefits. Indeed, the overwhelming majority of clinical studies in the field have been performed with viable probiotics. However, it has also been speculated that some of the mechanisms behind the probiotic health effects may not be dependent on the viability of the cells and, therefore, is also possible that also non-viable probiotics could have some health benefits. The efficacy of non-viable probiotics has been assessed in a limited number of studies, with varying success. While it is clear that viable probiotics are more effective than non-viable probiotics and that, in many cases, viability is indeed a prerequisite for the health benefit, there are also some cases where it appears that non-viable probiotics could also have beneficial effects on human health.

  18. ELECTRON-MICROSCOPIC ANALYSIS AND BIOCHEMICAL-CHARACTERIZATION OF A NOVEL METHANOL DEHYDROGENASE FROM THE THERMOTOLERANT BACILLUS-SP C1

    NARCIS (Netherlands)

    Vonck, Janet; Arfman, Nico; de Vries, Gert E.; Beeumen, Jozef van; Bruggen, Ernst F.J. van; Dijkhuizen, Lubbert

    1991-01-01

    Methanol dehydrogenase from the thermotolerant Bacillus sp. C1 was studied by electron microscopy and image processing. Two main projections can be distinguished: one exhibits 5-fold symmetry and has a diameter of 15 nm, the other is rectangular with sides of 15 and 9 nm. Subsequent image processing

  19. Toward an international standard for PCR-based detection of food-borne thermotolerant campylobacters: Validation in a multicenter collaborative trial

    DEFF Research Database (Denmark)

    Lübeck, Peter Stephensen; Cook, N.; Wagner, M.

    2003-01-01

    As part of a European research project, the performance of a PCR assay to detect food-borne thermotolerant campylobacters (Campylobacter jejuni, C. coli, and C. lari) was evaluated through an international collaborative trial involving 12 participating laboratories. DNA from 10 target and 8...

  20. Evaluation of data from the literature on the transport and survival of Escherichia coli and thermotolerant coliforms in aquifers under saturated conditions.

    NARCIS (Netherlands)

    Foppen, J W A; Schijven, J F

    2006-01-01

    Escherichia coli and thermotolerant coliforms are of major importance as indicators of fecal contamination of water. Due to its negative surface charge and relatively low die-off or inactivation rate coefficient, E. coli is able to travel long distances underground and is therefore also a useful

  1. Fungal Spores Viability on the International Space Station

    Science.gov (United States)

    Gomoiu, I.; Chatzitheodoridis, E.; Vadrucci, S.; Walther, I.; Cojoc, R.

    2016-11-01

    In this study we investigated the security of a spaceflight experiment from two points of view: spreading of dried fungal spores placed on the different wafers and their viability during short and long term missions on the International Space Station (ISS). Microscopic characteristics of spores from dried spores samples were investigated, as well as the morphology of the colonies obtained from spores that survived during mission. The selected fungal species were: Aspergillus niger, Cladosporium herbarum, Ulocladium chartarum, and Basipetospora halophila. They have been chosen mainly based on their involvement in the biodeterioration of different substrate in the ISS as well as their presence as possible contaminants of the ISS. From biological point of view, three of the selected species are black fungi, with high melanin content and therefore highly resistant to space radiation. The visual inspection and analysis of the images taken before and after the short and the long term experiments have shown that all biocontainers were returned to Earth without damages. Microscope images of the lids of the culture plates revealed that the spores of all species were actually not detached from the surface of the wafers and did not contaminate the lids. From the adhesion point of view all types of wafers can be used in space experiments, with a special comment on the viability in the particular case of iron wafers when used for spores that belong to B. halophila (halophilic strain). This is encouraging in performing experiments with fungi without risking contamination. The spore viability was lower in the experiment for long time to ISS conditions than that of the short experiment. From the observations, it is suggested that the environment of the enclosed biocontainer, as well as the species'specific behaviour have an important effect, reducing the viability in time. Even the spores were not detached from the surface of the wafers, it was observed that spores used in the

  2. On the Breeding of Bivoltine Breeds of the Silkworm, Bombyx mori L. (Lepidoptera: Bombycidae, Tolerant to High Temperature and High Humidity Conditions of the Tropics

    Directory of Open Access Journals (Sweden)

    Harjeet Singh

    2010-01-01

    Full Text Available The hot climatic conditions of tropics prevailing particularly in summer are contributing to the poor performance of the bivoltine breeds and the most important aspect is that many quantitative characters such as viability and cocoon traits decline sharply when temperature is high. Hence, in a tropical country like India, it is very essential to develop bivoltine breeds/hybrids which can withstand the high temperature stress conditions. This has resulted in the development of CSR18 × CSR19, compatible hybrid for rearing throughout the year by utilizing Japanese thermotolerant hybrids as breeding resource material. Though, the introduction of CSR18 × CSR19 in the field during summer months had considerable impact, the productivity level and returns realized do not match that of other productive CSR hybrids. Therefore, the acceptance level of this hybrid with the farmers was not up to the expected level. This has necessitated the development of a temperature tolerant hybrid with better productivity traits than CSR18 × CSR19. Though, it was a difficult task to break the negative correlation associated with survival and productivity traits, attempts on this line had resulted in the development of CSR46 × CSR47, a temperature tolerant bivoltine hybrid with better productivity traits than CSR18 × CSR19. However, though, these hybrids are tolerant to high temperature environments, they are not tolerant to many of the silkworm diseases. Keeping this in view, an attempt is made to develop silkworm hybrids tolerant to high temperature environments.

  3. Characterization of a Thermotolerant Phytase Produced by Rhizopus microsporus var. microsporus Biofilm on an Inert Support Using Sugarcane Bagasse as Carbon Source.

    Science.gov (United States)

    Sato, Vanessa Sayuri; Jorge, João Atílio; Guimarães, Luis Henrique Souza

    2016-06-01

    The Rhizopus microsporus var. microsporus biofilm was able to produce increased levels of an extracellular thermotolerant phytase using polyethylene and viscose as an inert support in both modified NBRIP medium and modified Khanna medium containing sugarcane bagasse as the carbon source. The enzyme production was strictly regulated by the phosphorus content with optimal production at 0.5 mM of sodium phytate and KH2PO4. The extracellular phytase, RMPhy1, was purified 4.18-fold with 4.78 % recovery using DEAE-cellulose and CM-cellulose. A single protein band with a molecular mass of 35.4 kDa was obtained when the samples were subjected to 10 % SDS-PAGE. The optimum temperature for activity was 55 °C and the optimum pH was 4.5. R. microsporus var. microsporus phytase exhibited high stability at 30 and 40 °C with a half-life of 115 min at 60 °C. The enzyme activity increased in the presence of Ca (2+) and was inhibited by Zn(2+), arsenate, and sodium phosphate. Phytase demonstrated high substrate specificity for sodium phytate with K m = 0.72 mM and V max = 94.55 U/mg of protein and for p-NPP with K m = 0.04 mM and V max = 106.38 U/mg of protein. The enzyme also hydrolyzed ATP, AMPc, glucose 6-phosphate, glucose 1-phosphate, and UDPG. This is the first report on phytase characterization delivered with biofilm technology. The properties of the enzyme account for its high potential for use in biotechnology and the possibility of application in different industrial sectors as feed in the future.

  4. [Hemodialysis improves the subendocardial viability ratio].

    Science.gov (United States)

    De Blasio, Antonella; Sirico, Maria; Di Micco, Lucia; Di Iorio, Biagio

    2013-01-01

    The subendocardial viability ratio (SEVR), a parameter introduced by Buckberg, represents a non-invasive measure of myocardial perfusion related to left ventricular work. AIM. The aim of this study was to verify if dialysis may determine modifications of SEVR and how these modifications are modulated in the 2-day interdialytic period. METHODS.We studied 54 subjects of mean age 6314 years and receiving dialysis for 3215 months. Exclusion criteria were diabetes, resistant hypertension and peripheral vascular diseases and intradialytic hypotension evidenced during the study dialysis session. Pulse wave velocity and SEVR assessments were performed during the third dialysis session of the week, before (pre-HD) and after (post-HD) dialysis, in 2-day interdialytic period after and at the beginning of the following dialysis session. RESULTS.Dialysis reduces PWV, in particular the tertile with the lowest PWV presents the highest percentage reduction (-26%) compared with the second and the third tertiles. In the same way, dialysis leads to an increase of SEVR and patients in the tertile with the highest SEVR values maintain high SEVR values during dialysis and in the interdialytic period. Patients with severe vascular calcifications present higher PWV value and lower SEVR value. CONCLUSIONS.The results of present study demonstrate that ultrafiltration improves PWV (with a mean reduction of 16%) and SEVR (increase of 13%) and that the severity of vascular calcifications influences the effect of ultrafiltration on these two parameters. More studies are certainly necessary to verify our findings. Considered the higher mortality of patients with higher SEVR, it would be important to understand if new dialytic strategies are needed in patients with higher PVW and lower SEVR values.

  5. Nuclear Power Options Viability Study. Volume 4. Bibliography

    Energy Technology Data Exchange (ETDEWEB)

    Trauger, D B; White, J D; Sims, J W [eds.

    1986-09-01

    Documents in the Nuclear Power Options Viability Study (NPOVS) bibliography are classified under one of four headings or categories as follows: nuclear options; light water reactors; liquid metal reactors; and high temperature reactors. The collection and selection of these documents, beginning early in 1984 and continuing through March of 1986, was carried out in support of the study's objective: to explore the viabilities of several nuclear electric power generation options for commercial deployment in the United States between 2000 and 2010. There are approximately 550 articles, papers, reports, and books in the bibliography that have been selected from some 2000 surveyed. The citations have been made computer accessible to facilitate rapid on-line retrieval by keyword, author, corporate author, title, journal name, or document number.

  6. Different effects of sonoporation on cell morphology and viability

    Directory of Open Access Journals (Sweden)

    Ji-Zhen Zhang

    2012-05-01

    Full Text Available The objective of our study was to investigate changes in cell morphology and viability after sonoporation. Sonoportion was achieved by ultrasound (21 kHz exposure on adherent human prostate cancer DU145 cells in the cell culture dishes with the presence of microbubble contrast agents and calcein (a cell impermeant dye. We investigated changes in cell morphology immediately after sonoporation under scanning electron microscope (SEM and changes in cell viability immediately and 6 h after sonoporation under fluorescence microscope. It was shown that various levels of intracellular calcein uptake and changes in cell morphology can be caused immediately after sonoporation: smooth cell surface, pores in the membrane and irregular cell surface. Immediately after sonoporation, both groups of cells with high levels of calcein uptake and low levels of calcein uptake were viable; 6 h after sonoporation, group of cells with low levels of calcein uptake still remained viable, while group of cells with high levels of calcein uptake died. Sonoporation induces different effects on cell morphology, intracellular calcein uptake and cell viability.

  7. The Effectiveness of Sanitary Inspections as a Risk Assessment Tool for Thermotolerant Coliform Bacteria Contamination of Rural Drinking Water: A Review of Data from West Bengal, India.

    Science.gov (United States)

    Snoad, Christian; Nagel, Corey; Bhattacharya, Animesh; Thomas, Evan

    2017-04-01

    AbstractThe use of sanitary inspections combined with periodic water quality testing has been recommended in some cases as screening tools for fecal contamination. We conducted sanitary inspections and tested for thermotolerant coliforms (TTCs), a fecal indicator bacteria, among 7,317 unique water sources in West Bengal, India. Our results indicate that the sanitary inspection score has poor ability to identify TTC-contaminated sources. Among deep and shallow hand pumps, the area under curve (AUC) for prediction of TTC > 0 was 0.58 (95% confidence interval [CI] = 0.53-0.61) and 0.58 (95% CI = 0.54-0.62), respectively, indicating that the sanitary inspection score was only marginally better than chance in discriminating between contaminated and uncontaminated sources of this type. A slightly higher AUC value of 0.64 (95% CI=0.57-0.71) was observed when the sanitary inspection score was used for prediction of TTC > 0 among the gravity-fed piped sources. Among unprotected springs (AUC = 0.48, 95% CI = 0.38-0.55) and unprotected dug wells (AUC = 0.41, 95% CI = 0.20-0.66), the sanitary inspection score performed more poorly than chance in discriminating between sites with TTC 0. Aggregating over all source types, the sensitivity (true positive rate) of a high/very high sanitary inspection score for TTC contamination (TTC > 1 CFU/100 mL) was 29.4% and the specificity (true negative rate) was 77.9%, resulting in substantial misclassification of the sites when using the established risk categories. These findings suggest that sanitary surveys are inappropriate screening tools for identifying TTC contamination at water points.

  8. Engineering of xylose reductase and overexpression of xylitol dehydrogenase and xylulokinase improves xylose alcoholic fermentation in the thermotolerant yeast Hansenula polymorpha

    Directory of Open Access Journals (Sweden)

    Voronovsky Andriy Y

    2008-07-01

    Full Text Available Abstract Background The thermotolerant methylotrophic yeast Hansenula polymorpha is capable of alcoholic fermentation of xylose at elevated temperatures (45 – 48°C. Such property of this yeast defines it as a good candidate for the development of an efficient process for simultaneous saccharification and fermentation. However, to be economically viable, the main characteristics of xylose fermentation of H. polymorpha have to be improved. Results Site-specific mutagenesis of H. polymorpha XYL1 gene encoding xylose reductase was carried out to decrease affinity of this enzyme toward NADPH. The modified version of XYL1 gene under control of the strong constitutive HpGAP promoter was overexpressed on a Δxyl1 background. This resulted in significant increase in the KM for NADPH in the mutated xylose reductase (K341 → R N343 → D, while KM for NADH remained nearly unchanged. The recombinant H. polymorpha strain overexpressing the mutated enzyme together with native xylitol dehydrogenase and xylulokinase on Δxyl1 background was constructed. Xylose consumption, ethanol and xylitol production by the constructed strain were determined for high-temperature xylose fermentation at 48°C. A significant increase in ethanol productivity (up to 7.3 times was shown in this recombinant strain as compared with the wild type strain. Moreover, the xylitol production by the recombinant strain was reduced considerably to 0.9 mg × (L × h-1 as compared to 4.2 mg × (L × h-1 for the wild type strain. Conclusion Recombinant strains of H. polymorpha engineered for improved xylose utilization are described in the present work. These strains show a significant increase in ethanol productivity with simultaneous reduction in the production of xylitol during high-temperature xylose fermentation.

  9. Economic Viability and Marketing Strategies of Periwinkle ...

    African Journals Online (AJOL)

    The economic viability and marketing strategies of periwinkle species in twelve major markets across Rivers State Nigeria were investigated using structured questionnaires. The results indicated that marketing strategies are enroute, through harvesters (collectors), to wholesalers (those who purchase in small quantities ...

  10. Extending the viability of sea urchin gametes.

    Science.gov (United States)

    Spiegler, M A; Oppenheimer, S B

    1995-04-01

    The sea urchin is the material of choice for studying many early developmental events. Methods to extend the viability of sea urchin gametes have not received much attention, but it is well known that the eggs are easily damaged by freezing. This study was designed to extend the viability of Lytechinus pictus eggs and sperm without freezing. Gamete viability measurements were based on relative numbers of fertilized vs unfertilized eggs, percentage fertilization, and on observations of embryonic development. Results indicate that gametes can be stored longer and at lower temperatures than previously described. Sperm were consistently kept viable for at least 12 days with little decrease in viability when stored in glass test tubes or plastic petri dishes and submerged in ice inside a refrigerator at 0 +/- 1 degree C. In one experiment, sperm stored in glass test tubes on ice remained viable up to 20 days after extraction. Eggs were maintained from 1 to 7 days, rather than the 1 day or so previously reported, when stored in glass test tubes submerged in ice in a refrigerator at 0 +/- 1 degree C. Results of egg and sperm experiments varied at different times in the season. Such variations may be caused by seasonal cytoplasmic changes, population differences, or the time mature individuals were maintained unfed in aquaria prior to use. Results from this study should be useful for a variety of research, mariculture, and teaching applications in which sea urchin supplies are limited or when the same gamete population is required for subsequent experiments.

  11. Incorporating evolutionary processes into population viability models

    NARCIS (Netherlands)

    Pierson, J.C.; Beissinger, S.R.; Bragg, J.G.; Coates, D.J.; Oostermeijer, J.G.B.; Sunnucks, P.; Schumaker, N.H.; Trotter, M.V.; Young, A.G.

    2015-01-01

    We examined how ecological and evolutionary (eco-evo) processes in population dynamics could be better integrated into population viability analysis (PVA). Complementary advances in computation and population genomics can be combined into an eco-evo PVA to offer powerful new approaches to understand

  12. Assessment of myocardial viability using PET

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Seok Nam [College of Medicine, Ajou University, Suwon (Korea, Republic of)

    2005-02-15

    The potential for recovery of left ventricular dysfunction after myocardial revascularization represents a practical clinical definition for myocardial viability. The evaluation of viable myocardium in patients with severe global left ventricular dysfunction due to coronary artery disease and with regional dysfunction after acute myocardial infarction is an important issue whether left ventricular dysfunction may be reversible or irreversible after therapy. If the dysfunction is due to stunning or hibernation, functional improvement is observed. But stunned myocardium may recover of dysfunction with no revascularization. Hibernation is chronic process due to chronic reduction in the resting myocardial blood flow. There are two types of myocardial hibernation; 'functional hibernation' with preserved contractile reserve and 'structural hibernation' without contractile reserve in segments with preserved glucose metabolism. This review focus on the application of F-18 FDG and other radionuclides to evaluate myocardial viability. In addition the factors influencing predictive value of FDG imaging for evaluating viability and the different criteria for viability are also reviewed.

  13. Pollen viability in Quercus robur L.

    Directory of Open Access Journals (Sweden)

    Batos Branislava

    2017-01-01

    Full Text Available The variability of viability (germination rate and the length of pollen tubes of fresh pedunculate oak (Quercus robur L. pollen grains was studied in vitro on a medium containing 15% sucrose. Spatial variability was studied by sampling fresh pollen grains from a total of thirteen trees at four different sites in the area of Belgrade (Košutnjak, Banovo Brdo, Ada Ciganlija and Bojčin Forest in a single year (2011. In order to assess temporal variability and determine the effects of climate change on a small time scale, we studied the viability of the pollen grains collected from one tree at the Banovo Brdo site in six different years (2004, 2005, 2006, 2007, 2011 and 2012. Interindividual variability was tested on the pollen grains sampled from eight trees at Ada Ciganlija in 2004. The percentage values of the pollen grain germination rate and the pollen tube length showed no statistically significant differences between the sites. However, the studied characteristics of the pollen grain viability (germination rate and pollen tube length showed statistically significant differences in both temporal (between the pollen collection years and interindividual variability. This type of research makes a valuable contribution to pedunculate oak breeding programs through the identification of trees with stable production and a good quality of pollen. Furthermore, it can be important in defining the patterns of spatial, temporal and individual variability of pollen grain viability under the influence of climate factors, which are showing compelling changing trends from year to year.

  14. Viability of lactobacillus acidophilus in various vaginal tablet formulations

    Directory of Open Access Journals (Sweden)

    Fazeli M.R.

    2006-07-01

    Full Text Available The lactobacilli which are present in vaginal fluids play an important role in prevention of vaginosis and there are considerable interests in formulation of these friendly bacteria into suitable pharmaceutical dosage forms. Formulating these microorganisms for vaginal application is a critical issue as the products should retain viability of lactobacilli during formulation and also storage. The aim of this study was to examine the viability and release of Lactobacillus acidophilus from slow-release vaginal tablets prepared by using six different retarding polymers and from two effervescent tablets prepared by using citric or adipic acid. The Carbomer–based formulations showed high initial viablility compared to those based on HPMC-LV, HPMC-HV, Polycarbophil and SCMC polymers which showed one log decrease in viable cells. All retarding polymers in slow release formulations presented a strong bacterial release at about 2 h except Carbomer polymers which showed to be poor bacterial releasers. Although effervescent formulations produced a quick bacterial release in comparison with polymer based slow-release tablets, they were less stable in cold storage. Due to the strong chelating characteristic of citric acid, the viability was quickly lost for aqueous medium of citric acid in comparison with adipic acid based effervescent tablets.

  15. PET/SPECT imaging: From carotid vulnerability to brain viability

    Energy Technology Data Exchange (ETDEWEB)

    Meerwaldt, Robbert [Department of Surgery, Isala Clinics, Zwolle (Netherlands); Slart, Riemer H.J.A. [Department of Nuclear Medicine and Molecular Imaging, University Medical Center Groningen, Groningen (Netherlands); Dam, Gooitzen M. van [Department of Surgery, University Medical Center Groningen, Groningen (Netherlands); Luijckx, Gert-Jan [Department of Neurology, University Medical Center Groningen, Groningen (Netherlands); Tio, Rene A. [Department of Cardiology, University Medical Center Groningen, Groningen (Netherlands); Zeebregts, Clark J. [Department of Surgery, University Medical Center Groningen, Groningen (Netherlands)], E-mail: czeebregts@hotmail.com

    2010-04-15

    Background: Current key issues in ischemic stroke are related to carotid plaque vulnerability, brain viability, and timing of intervention. The treatment of ischemic stroke has evolved into urgent active interventions, as 'time is brain'. Functional imaging such as positron emission tomography (PET)/single photon emission computed tomography (SPECT) could improve selection of patients with a vulnerable plaque and evaluation of brain viability in ischemic stroke. Objective: To describe the current applications of PET and SPECT as a diagnostic tool in relation to ischemic stroke. Methods: A literature search using PubMed identified articles. Manual cross-referencing was also performed. Results: Several papers, all observational studies, identified PET/SPECT to be used as a tool to monitor systemic atheroma modifying treatment and to select high-risk patients for surgery regardless of the degree of luminal stenosis in carotid lesions. Furthermore, PET/SPECT is able to quantify the penumbra region during ischemic stroke and in this way may identify those patients who may benefit from timely intervention. Discussion: Functional imaging modalities such as PET/SPECT may become important tools for risk-assessment and evaluation of treatment strategies in carotid plaque vulnerability and brain viability. Prospective clinical studies are needed to evaluate the diagnostic accuracy of PET/SPECT.

  16. Genome sequence of thermotolerant Bacillus methanolicus: features and regulation related to methylotrophy and production of L-lysine and L-glutamate from methanol.

    Science.gov (United States)

    Heggeset, Tonje M B; Krog, Anne; Balzer, Simone; Wentzel, Alexander; Ellingsen, Trond E; Brautaset, Trygve

    2012-08-01

    Bacillus methanolicus can utilize methanol as its sole carbon and energy source, and the scientific interest in this thermotolerant bacterium has focused largely on exploring its potential as a biocatalyst for the conversion of methanol into L-lysine and L-glutamate. We present here the genome sequences of the important B. methanolicus model strain MGA3 (ATCC 53907) and the alternative wild-type strain PB1 (NCIMB13113). The physiological diversity of these two strains was demonstrated by a comparative fed-batch methanol cultivation displaying highly different methanol consumption and respiration profiles, as well as major differences in their L-glutamate production levels (406 mmol liter(-1) and 11 mmol liter(-1), respectively). Both genomes are small (ca 3.4 Mbp) compared to those of other related bacilli, and MGA3 has two plasmids (pBM19 and pBM69), while PB1 has only one (pBM20). In particular, we focus here on genes representing biochemical pathways for methanol oxidation and concomitant formaldehyde assimilation and dissimilation, the important phosphoenol pyruvate/pyruvate anaplerotic node, the tricarboxylic acid cycle including the glyoxylate pathway, and the biosynthetic pathways for L-lysine and L-glutamate. Several unique findings were made, including the discovery of three different methanol dehydrogenase genes in each of the two B. methanolicus strains, and the genomic analyses were accompanied by gene expression studies. Our results provide new insight into a number of peculiar physiological and metabolic traits of B. methanolicus and open up possibilities for system-level metabolic engineering of this bacterium for the production of amino acids and other useful compounds from methanol.

  17. Optimization of thermostable organic solvent-tolerant lipase production by thermotolerant Rhizopus sp. using solid-state fermentation of palm kernel cake.

    Science.gov (United States)

    Riyadi, Fatimah Azizah; Alam, Md Zahangir; Salleh, Md Noor; Salleh, Hamzah Mohd

    2017-10-01

    This study enhanced the production of thermostable organic solvent-tolerant (TS-OST) lipase by locally isolated thermotolerant Rhizopus sp. strain using solid-state fermentation (SSF) of palm kernel cake (PKC). The optimum conditions were achieved using a series of statistical approaches. The cultivation parameters, which include fermentation time, moisture content, temperature, pH, inoculum size, various carbon and nitrogen sources, as well as other supplements, were initially screened by the definitive screening design, and one-factor-at-a-time using PKC as the basal medium. Three significant factors (olive oil concentration, pH, and inoculum size) were further optimized using face-centred central composite design. The results indicated a successful and significant improvement of lipase activity by almost two-fold compared to the initial screening production. The findings showed that the optimal conditions were 2% (v/w) inoculum size, 2% (v/w) olive oil, 0.6% (w/w) peptone, 2% (v/w) ethanol, 70% moisture content at initial pH 10.0 and 45 °C within 72 h of fermentation. Process optimization resulted in maximum lipase activity of 58.63 U/gram dry solids (gds). The analysis of variance showed that the statistical model was significant (p value <0.0001) and reliable with a high value of R2 (0.98) and adjusted R2 (0.96). This indicates a better correlation between the actual and predicted responses of lipase production. By considering this study, the low-cost PKC through SSF appears to be promising in the utilization of agro-industrial waste for TS-OST lipase production. This is because satisfactory enzyme activity could be attained that promises industrial applications.

  18. Production of poly-γ-glutamic acid by a thermotolerant glutamate-independent strain and comparative analysis of the glutamate dependent difference.

    Science.gov (United States)

    Zeng, Wei; Chen, Guiguang; Guo, Ye; Zhang, Bin; Dong, Mengna; Wu, Yange; Wang, Jun; Che, Zhiqun; Liang, Zhiqun

    2017-11-25

    Poly-γ-glutamic acid (γ-PGA) is a promising microbial polymer with wide applications in industry, agriculture and medicine. In this study, a novel glutamate-independent γ-PGA producing strain with thermotolerant characteristics was isolated and identified as Bacillus subtilis GXG-5, then its product was also characterized. The fermentation process was optimized by single-factor tests, and results showed that high temperature (50 °C) was especially suitable for the γ-PGA production by GXG-5. The γ-PGA yield reached 19.50 ± 0.75 g/L with substrate conversion efficiency of 78% at 50 °C in 10 L fermentor. Comparison of GXG-5 and GXA-28 (glutamate-dependent strain) under respective optimal fermentation conditions, the γ-PGA yield of GXG-5 was 19.0% higher than that of GXA-28, and GXG-5 was also superior to GXA-28 in the availability of carbon sources and substrates. Furthermore, the glutamate dependent difference between GXA-28 and GXG-5 was analyzed by genomic sequencing, results indicated that genes related to the glutamate dependent difference mainly involved in carbohydrate transport and metabolism and amino acid metabolism, and 13 genes related to γ-PGA synthesis were mutated in GXG-5. This study provided a potential glutamate-independent strain to replace glutamate-dependent strain for γ-PGA production, and shared novel information for understanding the glutamate dependent difference at the genomic level.

  19. Data for rapid ethanol production at elevated temperatures by engineered thermotolerant Kluyveromyces marxianus via the NADP(H-preferring xylose reductase–xylitol dehydrogenase pathway

    Directory of Open Access Journals (Sweden)

    Biao Zhang

    2015-12-01

    Full Text Available A thermo-tolerant NADP(H-preferring xylose pathway was constructed in Kluyveromyces marxianus for ethanol production with xylose at elevated temperatures (Zhang et al., 2015 [25]. Ethanol production yield and efficiency was enhanced by pathway engineering in the engineered strains. The constructed strain, YZJ088, has the ability to co-ferment glucose and xylose for ethanol and xylitol production, which is a critical step toward enabling economic biofuel production from lignocellulosic biomass. This study contains the fermentation results of strains using the metabolic pathway engineering procedure. The ethanol-producing abilities of various yeast strains under various conditions were compared, and strain YZJ088 showed the highest production and fastest productivity at elevated temperatures. The YZJ088 xylose fermentation results indicate that it fermented well with xylose at either low or high inoculum size. When fermented with an initial cell concentration of OD600=15 at 37 °C, YZJ088 consumed 200 g/L xylose and produced 60.07 g/L ethanol; when the initial cell concentration was OD600=1 at 37 °C, YZJ088 consumed 98.96 g/L xylose and produced 33.55 g/L ethanol with a productivity of 0.47 g/L/h. When fermented with 100 g/L xylose at 42 °C, YZJ088 produced 30.99 g/L ethanol with a productivity of 0.65 g/L/h, which was higher than that produced at 37 °C.

  20. Ethanol Production Kinetics by a Thermo-Tolerant Mutant of Saccharomyces Cerevisiae from Starch Industry Waste (Hydrol

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    Farman Ali Shah

    2010-06-01

    Full Text Available A thermo-tolerant and deoxyglucose-resistant mutant of Saccharomyces cerevisiae was developed and employed to convert them to fuel ethanol in a 150 litre fermenter. Maximum ethanol production was achieved when fermentation of dextrozyme- treated hydrol was carried out for about 36 hours under optimized fermenting conditions. The maximum specific ethanol production rate (qP, and overall ethanol yield (YP/S were found to be 2.82 g L-1 h-1 and 0.49 g/g respectively. Determination of activation energy for cell growth (Ea= 20.8 kJ/mol and death (Ed = 19.1 kJ/mol and product formation and inactivation (EP=35.8 kJ/mol and Edp = 33.5 kJ/mol revealed the thermo-stability of the organism for up to 47°C.

  1. Adenovirus, enterovirus and thermotolerant coliforms in recreational waters from Lake Guaíba beaches, Porto Alegre, Brazil.

    Science.gov (United States)

    Maurer, C P; Simonetti, A B; Staggemeier, R; Rigotto, C; Heinzelmann, L S; Spilki, F R

    2015-12-01

    In the present study, molecular detection of human adenoviruses (HAdV) and enteroviruses (EV) was performed in surface water samples collected from beaches Ipanema and Lami, located on the shores of Lake Guaíba, city of Porto Alegre, RS, southern Brazil. Furthermore, water safety was evaluated by counting thermotolerant coliforms (TC), following local government regulations. A total of 36 samples were collected monthly from six different sites along the beaches. Viral genomes were found in 30 (83.3%) samples. The higher detection rate was observed for HAdV (77.8%), followed by EV (22.2%). Although low concentrations of TC have been found, the occurrence of viral genomes in water samples was frequent and may pose a potential risk of infection for people bathing in these beaches.

  2. Monitoring cell growth, viability, and apoptosis.

    Science.gov (United States)

    Butler, Michael; Spearman, Maureen; Braasch, Katrin

    2014-01-01

    The accurate determination of cell growth and viability is pivotal to monitoring a bioprocess. Direct methods to determine the cell growth and/or viability in a bioprocess include microscopic counting, electronic particle counting, image analysis, in situ biomass monitoring, and dieletrophoretic cytometry. These methods work most simply when a fixed volume sample can be taken from a suspension culture. Manual microscopic counting is laborious but affords the advantage of allowing cell viability to be determined if a suitable dye is included. Electronic particle counting is a rapid total cell count method for replicate samples, but some data distortion may occur if the sample has significant cell debris or cell aggregates. Image analysis based on the use of digital camera images acquired through a microscope has advanced rapidly with the availability of several commercially available software packages replacing manual microscopic counting and viability determination. Biomass probes detect cells by their dielectric properties or their internal concentration of NADH and can be used as a continuous monitor of the progress of a culture. While the monitoring of cell growth and viability is an integral part of a bioprocess, the monitoring of apoptosis induction is also becoming more and more important in bioprocess control to increase volumetric productivity by extending bioprocess duration. Different fluorescent assays allow for the detection of apoptotic characteristics in a cell sample.Indirect methods of cell determination involve the chemical analysis of a culture component or a measure of metabolic activity. These methods are most useful when it is difficult to obtain intact cell samples. However, the relationship between these parameters and the cell number may not be linear through the phases of a cell culture. The determination of nucleic acid (DNA) or total protein can be used as an estimate of biomass, while the depletion of glucose from the media can be used

  3. No. 347-Obstetric Management at Borderline Viability.

    Science.gov (United States)

    Ladhani, Noor Niyar N; Chari, Radha S; Dunn, Michael S; Jones, Griffith; Shah, Prakesh; Barrett, Jon F R

    2017-09-01

    The primary objective of this guideline was to develop consensus statements to guide clinical practice and recommendations for obstetric management of a pregnancy at borderline viability, currently defined as prior to 25+6 weeks. Clinicians involved in the obstetric management of women whose fetus is at the borderline of viability. Women presenting for possible birth at borderline viability. This document presents a summary of the literature and a general consensus on the management of pregnancies at borderline viability, including maternal transfer and consultation, administration of antenatal corticosteroids and magnesium sulfate, fetal heart rate monitoring, and considerations in mode of delivery. Medline, EMBASE, and Cochrane databases were searched using the following keywords: extreme prematurity, borderline viability, preterm, pregnancy, antenatal corticosteroids, mode of delivery. The results were then studied, and relevant articles were reviewed. The references of the reviewed studies were also searched, as were documents citing pertinent studies. The evidence was then presented at a consensus meeting, and statements were developed. The content and recommendations were developed by the consensus group from the fields of Maternal-Fetal Medicine, Neonatology, Perinatal Nursing, Patient Advocacy, and Ethics. The quality of evidence was rated using criteria described in the Grading of Recommendations Assessment, Development and Evaluation methodology framework (reference 1). The Board of the Society of Obstetricians and Gynaecologists of Canada approved the final draft for publication. The quality of evidence was rated using the criteria described in the Grading of Recommendations, Assessment, Development, and Evaluation methodology framework. The interpretation of strong and weak recommendations is described later. The Summary of Findings is available upon request. A multidisciplinary approach should be used in counselling women and families at borderline

  4. On the evolution of genetic incompatibility systems. V. Origin of sporophytic self-incompatibility in response to overdominance in viability.

    Science.gov (United States)

    Uyenoyama, M K

    1989-12-01

    Conditions for the origin of partial sporophytic self-incompatibility (SSI) are obtained from two quantitative models, which differ with respect to the determination of offspring viability. Offspring viability depends solely on the source (self or nonself) of the fertilizing pollen in the first model, which describes changes only at a primitive S-locus itself. Two loci evolve in the second model: overdominant viability selection maintains an arbitrary number of alleles at one locus, with SSI under the control of a separate locus. In both cases, the origin of SSI requires that the relative change in the numbers of offspring derived by the two reproductive modes compensate for the twofold cost of outcrossing. In the first model studied, the viability of inbred offspring fully determines the relative change in the numbers of inbred and outbred offspring produced. In the second model, the relative change in offspring numbers depends in addition on associations between the S-locus and the viability locus. Because these two-locus associations are comparable in magnitude to the differences between the viabilities of inbred and outbred offspring, SSI can arise under less restrictive conditions than expected from the one-locus model. Greater allelic multiplicity at the viability locus facilitates the origin of SSI by reducing the relative viability of inbred offspring. Tight linkage between the S-locus and the viability locus and high rates of receipt of self-pollen promote the generation and maintenance of associations between the S-locus and the viability locus. In populations in which more than two viability alleles are maintained, the active S-allele can invade even in the absence of linkage with the viability locus. The present study establishes that incompatibility systems can arise in response to identity disequilibrium between a modifier of incompatibility and a locus subject to overdominant viability selection; in particular, compensation for the twofold cost of

  5. The Viability of Small Banks in the United States

    OpenAIRE

    R. Alton Gilbert

    2007-01-01

    Small banks have an important role in financing economic activity through their financial services for small businesses. There has been a sharp decline in the number of small banking organizations in the U.S. since the early 1980s. A continuation of this trend would raise important issues about access to financial services for small businesses. Data on the number of banks, their profits, and the distribution of consistent high and low earning banks tend to tell the same story about the viabil...

  6. A simple colony-formation assay in liquid medium, termed 'tadpoling', provides a sensitive measure of Saccharomyces cerevisiae culture viability.

    Science.gov (United States)

    Welch, Aaron Z; Koshland, Douglas E

    2013-12-01

    Here we describe the first high-throughput amenable method of quantifying Saccharomyces cerevisiae culture viability. Current high-throughput methods of assessing yeast cell viability, such as flow cytometry and SGA analysis, do not measure the percentage viability of a culture but instead measure cell vitality or colony fitness, respectively. We developed a method, called tadpoling, to quantify the percentage viability of a yeast culture, with the ability to detect as few as one viable cell amongst ~10(8) dead cells. The most important feature of this assay is the exploitation of yeast colony formation in liquid medium. Utilizing a microtiter dish, we are able to observe a range of viability of 100% to 0.0001%. Comparison of tadpoling to the traditional plating method to measure yeast culture viability reveals that, for the majority of Saccharomyces species analyzed there is no significant difference between the two methods. In comparison to flow cytometry using propidium iodide, the high-throughput method of measuring yeast culture viability, tadpoling is much more accurate at culture viabilities < 1%. Thus, we show that tadpoling provides an easy, inexpensive, space-saving method, amenable to high-throughput screens, for accurately measuring yeast cell viability. Copyright © 2013 John Wiley & Sons, Ltd.

  7. Soy milk as a storage medium to preserve human fibroblast cell viability: an in vitro study.

    Science.gov (United States)

    Moura, Camilla Christian Gomes; Soares, Priscilla Barbosa Ferreira; Reis, Manuella Verdinelli de Paula; Fernandes Neto, Alfredo Júlio; Soares, Carlos José

    2012-01-01

    Soy milk (SM) is widely consumed worldwide as a substitute for cow milk. It is a source of vitamins, carbohydrates and sugars, but its capacity to preserve cell viability has not been evaluated. The purpose of the present study was to investigate the efficacy of SM to maintain the viability of human fibroblasts at short periods compared with different cow milks. Human mouth fibroblasts were cultured and stored in the following media at room temperature: 10% Dulbecco's Modified Eagle Medium (DMEM) (positive control group); long shelf-life ultra-high temperature whole cow milk (WM); long shelf-life ultra-high temperature skim cow milk (SKM); powdered cow milk (PM); and soy milk (SM). After 5, 15, 30 and 45 min, cell viability was analyzed using the MTT assay. Data were analyzed statistically by the Kruskal-Wallis test with post-analysis using the Dunn's method (α=0.05). SKM showed the lowest capacity to maintain cell viability in all analyzed times (p<0.05). At 30 and 45 min, the absorbance levels in control group (DMEM) and SM were significantly higher than in SKM (p<0.05). Cell viability decreased along the time (5-45 min). The results indicate that SM can be used as a more adequate storage medium for avulsed teeth. SKM was not as effective in preserving cell viability as the cell culture medium and SM.

  8. Rapid onsite assessment of spore viability.

    Energy Technology Data Exchange (ETDEWEB)

    Branda, Steven; Lane, Todd W.; VanderNoot, Victoria A.; Gaucher, Sara P.; Jokerst, Amanda S.

    2005-12-01

    This one year LDRD addresses problems of threat assessment and restoration of facilities following a bioterror incident like the incident that closed down mail facilities in late 2001. Facilities that are contaminated with pathogenic spores such as B. anthracis spores must be shut down while they are treated with a sporicidal agent and the effectiveness of the treatment is ascertained. This process involves measuring the viability of spore test strips, laid out in a grid throughout the facility; the CDC accepted methodologies require transporting the samples to a laboratory and carrying out a 48 hr outgrowth experiment. We proposed developing a technique that will ultimately lead to a fieldable microfluidic device that can rapidly assess (ideally less than 30 min) spore viability and effectiveness of sporicidal treatment, returning facilities to use in hours not days. The proposed method will determine viability of spores by detecting early protein synthesis after chemical germination. During this year, we established the feasibility of this approach and gathered preliminary results that should fuel a future more comprehensive effort. Such a proposal is currently under review with the NIH. Proteomic signatures of Bacillus spores and vegetative cells were assessed by both slab gel electrophoresis as well as microchip based gel electrophoresis employing sensitive laser-induced fluorescence detection. The conditions for germination using a number of chemical germinants were evaluated and optimized and the time course of protein synthesis was ascertained. Microseparations were carried out using both viable spores and spores inactivated by two different methods. A select number of the early synthesis proteins were digested into peptides for analysis by mass spectrometry.

  9. Thermotolerance adaptation to human-modified habitats occurs in the native range of the invasive ant Wasmannia auropunctata before long-distance dispersal.

    Science.gov (United States)

    Foucaud, Julien; Rey, Olivier; Robert, Stéphanie; Crespin, Laurent; Orivel, Jérôme; Facon, Benoit; Loiseau, Anne; Jourdan, Hervé; Kenne, Martin; Masse, Paul Serge Mbenoun; Tindo, Maurice; Vonshak, Merav; Estoup, Arnaud

    2013-06-01

    Key evolutionary events associated with invasion success are traditionally thought to occur in the introduced, rather than the native range of species. In the invasive ant Wasmannia auropunctata, however, a shift in reproductive system has been demonstrated within the native range, from the sexual non-dominant populations of natural habitats to the clonal dominant populations of human-modified habitats. Because abiotic conditions of human- modified habitats are hotter and dryer, we performed lab experiments on workers from a set of native and introduced populations, to investigate whether these ecological and genetic transitions were accompanied by a change in thermotolerance and whether such changes occurred before establishment in the introduced range. Thermotolerance levels were higher in native populations from human-modified habitats than in native populations from natural habitats, but were similar in native and introduced populations from human-modified habitats. Differences in thermotolerance could not be accounted for by differences in body size. A scenario based on local adaptation in the native range before introduction in remote areas represents the most parsimonious hypothesis to account for the observed phenotypic pattern. These findings highlight the importance of human land use in explaining major contemporary evolutionary changes.

  10. Heat shock-triggered Ca2+ mobilization accompanied by pectin methylesterase activity and cytosolic Ca2+ oscillation are crucial for plant thermotolerance.

    Science.gov (United States)

    Wu, Hui-Chen; Jinn, Tsung-Luo

    2010-10-01

    Apoplastic Ca(2+) concentration controls membrane permeability, cell wall stabilization, and cell integrity; however, little is known about its role in thermotolerance in plants. Here, we report that the acquired thermotolerance of etiolated rice seedlings (Oryza sativa) was abolished by an exogenously supplied Ca(2+) chelator, EGTA, related to increased cellular content leakage during heat shock (HS) treatment. Thermotolerance was restored by the addition of Ca(2+) during EGTA incubation. Pectin methylesterase (EC 3.1.1.11), a cell-wall remodeling enzyme, was activated in response to HS, and its elevated activity was related to the recovery of the HS-released Ca(2+) concentration. EGTA interfered with the capability of HS to increase oscillation of [Ca(2+)]cyt content. We assume that heat-activated PME activity is involved in cell-wall-localized Ca(2+). The removal of apoplastic Ca(2+) might participate in HS signaling to induce HS protein expression and cell-wall remodeling to retain plasma membrane integrity, prevent cellular content leakage and confer thermoprotection. © 2010 Landes Bioscience

  11. Non-disruptive measurement system of cell viability in bioreactors

    Science.gov (United States)

    Rudek, F.; Nelsen, B. L.; Baselt, T.; Berger, T.; Wiele, M.; Prade, I.; Hartmann, P.

    2016-04-01

    Nutrient and oxygen transport, as well as the removal of metabolic waste are essential processes to support and maintain viable tissue. Current bioreactor technology used to grow tissue cultures in vitro has a fundamental limit to the thickness of tissues. Based on the low diffusion limit of oxygen a maximum tissue thickness of 200 μm is possible. The efficiency of those systems is currently under investigation. During the cultivation process of the artificial tissue in bioreactors, which lasts 28 days or longer, there are no possibilities to investigate the viability of cells. This work is designed to determine the influence of a non-disruptive cell viability measuring system on cellular activity. The measuring system uses a natural cellular marker produced during normal metabolic activity. Nicotinamide adenine dinucleotide (NADH) is a coenzyme naturally consumed and produced during cellular metabolic processes and has thoroughly been studied to determine the metabolic state of a cell. Measuring the fluorescence of NADH within the cell represents a non-disruptive marker for cell viability. Since the measurement process is optical in nature, NADH fluorescence also provides a pathway for sampling at different measurement depths within a given tissue sample. The measurement system we are using utilizes a special UV light source, to excite the NADH fluorescence state. However, the high energy potentially alters or harms the cells. To investigate the influence of the excitation signal, the cells were irradiated with a laser operating at a wavelength of 355 nm and examined for cytotoxic effects. The aim of this study was to develop a non-cytotoxic system that is applicable for large-scale operations during drug-tissue interaction testing.

  12. Cost Assessment Methodology and Economic Viability of Tidal Energy Projects

    Directory of Open Access Journals (Sweden)

    Eva Segura

    2017-11-01

    Full Text Available The exploitation of technologies with which to harness the energy from ocean currents will have considerable possibilities in the future thanks to their enormous potential for electricity production and their high predictability. In this respect, the development of methodologies for the economic viability of these technologies is fundamental to the attainment of a consistent quantification of their costs and the discovery of their economic viability, while simultaneously attracting investment in these technologies. This paper presents a methodology with which to determine the economic viability of tidal energy projects, which includes a technical study of the life-cycle costs into which the development of a tidal farm can be decomposed: concept and definition, design and development, manufacturing, installation, operation and maintenance and dismantling. These cost structures are additionally subdivided by considering their sub-costs and bearing in mind the main components of the tidal farm: the nacelle, the supporting tidal energy converter structure and the export power system. Furthermore, a technical study is developed in order to obtain an estimation of the annual energy produced (and, consequently, the incomes generated if the electric tariff is known by considering its principal attributes: the characteristics of the current, the ability of the device to capture energy and its ability to convert and export the energy. The methodology has been applied (together with a sensibility analysis to the particular case of a farm composed of first generation tidal energy converters in one of the Channel Island Races, the Alderney Race, in the U.K., and the results have been attained by means of the computation of engineering indexes, such as the net present value, the internal rate of return, the discounted payback period and the levelized cost of energy, which indicate that the proposed project is economically viable for all the case studies.

  13. Non-invasive imaging in detecting myocardial viability: Myocardial function versus perfusion

    Directory of Open Access Journals (Sweden)

    Iqbal A. Elfigih

    2014-12-01

    Full Text Available Coronary artery disease (CAD is the most prevalent and single most common cause of morbidity and mortality [1] with the resulting left ventricular (LV dysfunction an important complication. The distinction between viable and non-viable myocardium in patients with LV dysfunction is a clinically important issue among possible candidates for myocardial revascularization. Several available non-invasive techniques are used to detect and assess ischemia and myocardial viability. These techniques include echocardiography, radionuclide images, cardiac magnetic resonance imaging and recently myocardial computed tomography perfusion imaging. This review aims to distinguish between the available non-invasive imaging techniques in detecting signs of functional and perfusion viability and identify those which have the most clinical relevance in detecting myocardial viability in patients with CAD and chronic ischemic LV dysfunction. The most current available studies showed that both myocardial perfusion and function based on non-invasive imaging have high sensitivity with however wide range of specificity for detecting myocardial viability. Both perfusion and function imaging modalities provide complementary information about myocardial viability and no optimum single imaging technique exists that can provide very accurate diagnostic and prognostic viability assessment. The weight of the body of evidence suggested that non-invasive imaging can help in guiding therapeutic decision making in patients with LV dysfunction.

  14. Population Viability Analysis of Riverine Fishes

    Energy Technology Data Exchange (ETDEWEB)

    Bates, P.; Chandler, J.; Jager, H.I.; Lepla, K.; Van Winkle, W.

    1999-04-12

    Many utilities face conflkts between two goals: cost-efficient hydropower generation and protecting riverine fishes. Research to develop ecological simulation tools that can evaluate alternative mitigation strategies in terms of their benefits to fish populations is vital to informed decision-making. In this paper, we describe our approach to population viability analysis of riverine fishes in general and Snake River white sturgeon in particular. We are finding that the individual-based modeling approach used in previous in-stream flow applications is well suited to addressing questions about the viability of species of concern for several reasons. Chief among these are: (1) the abiIity to represent the effects of individual variation in life history characteristics on predicted population viabili~, (2) the flexibili~ needed to quanti~ the ecological benefits of alternative flow management options by representing spatial and temporal variation in flow and temperaturty and (3) the flexibility needed to quantifi the ecological benefits of non-flow related manipulations (i.e., passage, screening and hatchery supplementation).

  15. A New Methodology for Evaluation of Nematode Viability

    Directory of Open Access Journals (Sweden)

    Sebastião Rodrigo Ferreira

    2015-01-01

    Full Text Available Nematodes infections are responsible for debilitating conditions and economic losses in domestic animals as well as livestock and are considered an important public health problem due to the high prevalence in humans. The nematode resistance for drugs has been reported for livestock, highlighting the importance for development of new anthelmintic compounds. The aim of the current study was to apply and compare fluorimetric techniques using Sytox and propidium iodide for evaluating the viability of C. elegans larvae after treatment with anthelmintic drugs. These fluorescent markers were efficient to stain larvae treated with ivermectin and albendazole sulfoxide. We observed that densitometric values were proportional to the concentration of dead larvae stained with both markers. Furthermore, data on motility test presented an inverse correlation with fluorimetric data when ivermectin was used. Our results showed that lower concentrations of drugs were effective to interfere in the processes of cellular transport while higher drugs concentrations were necessary in order to result in any damage to cell integrity. The methodology described in this work might be useful for studies that aim to evaluate the viability of nematodes, particularly for testing of new anthelminthic compounds using an easy, economic, reproducible, and no time-consuming technique.

  16. Effect of various commercial buffers on sperm viability and capacitation.

    Science.gov (United States)

    Andrisani, Alessandra; Donà, Gabriella; Ambrosini, Guido; Bonanni, Guglielmo; Bragadin, Marcantonio; Cosmi, Erich; Clari, Giulio; Armanini, Decio; Bordin, Luciana

    2014-08-01

    A wide variety of sperm preparation protocols are currently available for assisted conception. They include density gradient separation and washing methods. Both aim at isolating and capacitating as much motile sperm as possible for subsequent oocyte fertilization. The aim of this study was to examine the effects of four commercial sperm washing buffers on sperm viability and capacitation. Semen samples from 48 healthy donors (normal values of sperm count, motility, morphology, and volume) were analyzed. After separation (density gradient 40/80%), sperm were incubated in various buffers then analysed for reactive oxygen species (ROS) production, viability, tyrosine phosphorylation (Tyr-P), cholera toxin B subunit (CTB) labeling, and the acrosome reaction (AR). The buffers affected ROS generation in various ways resulting either in rapid cell degeneration (when the amount of ROS was too high for cell survival) or the inability of the cells to maintain correct functioning (when ROS were too few). Only when the correct ROS generation curve was maintained, suitable membrane reorganization, evidenced by CTB labeling was achieved, leading to the highest percentages of both Tyr-P- and acrosome-reacted-cells. Distinguishing each particular pathological state of the sperm sample would be helpful to select the preferred buffer treatment since both ROS production and membrane reorganization can be significantly altered by commercial buffers.

  17. A Review: The Probiotic Bacteria Viability under Different Conditions

    Directory of Open Access Journals (Sweden)

    Lavinia Florina CALINOIU

    2016-11-01

    Full Text Available This review summarized the current knowledge on probiotics and on the effects that different conditions have under this type of bacteria. The purpose of this review was to evaluate the survival rate/resistance or viability of different probiotic bacteria under several conditions, such as: processing, food composition, storage, freezing, thawing, refrigeration, temperature, oxygen, pH, gastrointestinal environment and package. Nowadays, the demand on probiotic functional foods is increasing rapidly, as the consumers became more aware about the potential health benefits, due to the fact that probiotics help in maintaining the balance and composition of intestinal flora and protect it from pathogens. A daily ingestion of 108–109 CFU ml−1 probiotic microorganisms is crucial in order to be able to demonstrate an effect in our organism, considering the dose and the effect of storage/gastrointestinal environments on the probiotic viability. Microencapsulation of probiotics in different polysaccharides was proven to be an ideal way to preserve and protect the cells from detrimental factors during processing, storage or resistance in the gastrointestinal transit, as many studies demonstrate it. There is a general interest in the improvement of the physical and mechanical stability of the polymers used in probiotics encapsulation, to ensure high population of probiotics not only in food during storage, but also after gastrointestinal digestion. Also, the carrier plays a very important role and should be carefully examined.

  18. Evaluation of fermentation, drying, and high pressure processing on viability of Listeria monocytogenes, Escherichia coli O157:H7, Salmonella spp., and Trichinella spiralis in raw pork and/or Genoa salami

    Science.gov (United States)

    We evaluated the effectiveness of fermentation, drying, and high pressure processing (HPP) to inactivate Listeria monocytogenes, Escherichia coli O157:H7, Salmonella spp., and Trichinella spiralis in Genoa salami produced with trichinae infected pork. In addition, we evaluated the effectiveness of u...

  19. Pollen viability and its effect on fruit set of oil palm (Elaeis guineensis Jacq.

    Directory of Open Access Journals (Sweden)

    ALFIN WIDIASTUTI

    2008-01-01

    Full Text Available The research was aimed at studying (1 the decline of pollen viability during storage, and (2 the effect of pollen amount on fruit set of oil palm (Elaeis guineensis Jacq.. The experiment was conducted at PT. Dami Mas Sejahtera and PT. Sinar Mas Agro Resource and Technology (SMART Tbk, Riau from February to August 2004. The first experiment was investigated up to six months storage period in the refrigerator, whereas in the second experiment a randomized complete block design with two factors was used: length of storage, i.e. 0, 1 and 2 months and amount of pollen, i.e. 0.022, 0.044, 0.066, 0.088, and 0.11 g mixed with powder to 10g to pollinate an inflorescence. The result showed that the viability of pollen started to decline three months after storage from about 92% to 83%, and declined to about 75% after six months of storage. Result of the second experiment showed that storage of pollen up to two months did not affect percentage of normal fruit, although the percentage of parthenocarpic fruits was decreased. This could be due to the high viability of pollen as the viability was remained high (about 90% after being stored for two months in the refrigerator. Pollen with high viability could be used in a smaller amount to pollinate a female inflorescence without affecting fruit set of about 70-76%.SD037 had a higher reproductive success than SD038 and SD39.

  20. Exercise regulates breast cancer cell viability

    DEFF Research Database (Denmark)

    Dethlefsen, Christine; Lillelund, Christian; Midtgaard, Julie

    2016-01-01

    Purpose: Exercise decreases breast cancer risk and disease recurrence, but the underlying mechanisms are unknown. Training adaptations in systemic factors have been suggested as mediating causes. We aimed to examine if systemic adaptations to training over time, or acute exercise responses......, in breast cancer survivors could regulate breast cancer cell viability in vitro. Methods: Blood samples were collected from breast cancer survivors, partaking in either a 6-month training intervention or across a 2 h acute exercise session. Changes in training parameters and systemic factors were evaluated...... and pre/post exercise-conditioned sera from both studies were used to stimulate breast cancer cell lines (MCF-7, MDA-MB-231) in vitro. Results: Six months of training increased VO2peak (16.4 %, p

  1. Enhanced ethanol production from sugarcane juice by galactose adaptation of a newly isolated thermotolerant strain of Pichia kudriavzevii.

    Science.gov (United States)

    Dhaliwal, Sandeep Singh; Oberoi, Harinder Singh; Sandhu, Simranjeet Kaur; Nanda, Dhiraj; Kumar, Dinesh; Uppal, Satinder Kaur

    2011-05-01

    The thermotolerant yeast strain isolated from sugarcane juice through enrichment technique was identified as a strain of Pichiakudriavzevii (Issatchenkiaorientalis) through molecular characterization. The P. kudriavzevii cells adapted to galactose medium produced about 30% more ethanol from sugarcane juice than the non-adapted cells. The recycled cells could be used for four successive cycles without a significant drop in ethanol production. Fermentation in a laboratory fermenter with galactose adapted P. kudriavzevii cells at 40°C resulted in an ethanol concentration and productivity of 71.9 g L(-1) and 4.0 g L(-1)h(-1), respectively from sugarcane juice composed of about 14% (w/v) sucrose, 2% (w/v) glucose and 1% (w/v) fructose. In addition to ethanol, 3.30 g L(-1) arabitol and 4.19 g L(-1) glycerol were also produced, whereas sorbitol and xylitol were not formed during fermentation. Use of galactose adapted P. kudriavzevii cells for ethanol production from sugarcane juice holds potential for scale-up studies. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. Number of residual thermotolerant coliforms on plants and in soil when using reclaimed domestic wastewater for irrigation.

    Science.gov (United States)

    Khamkure, Sasirot; Cervantes, Edmundo Peña; Zermeño González, Alejandro; Cervantes, Rubén López; Melo, Prócoro Gamero; Ramírez, Homero

    2013-01-01

    The reclamation of domestic wastewater for irrigation is one alternative approach to solve the water scarcity crisis, but it is essential to control the microbiological quality of wastewater used for irrigation. The removal of thermotolerant coliforms, also known as faecal coliforms (FC), from treated domestic wastewater by intermittent media infiltration (IMI) in column was studied. The columns were filled with natural filter media (soil, soil/charcoal and zinc-modified zeolite, Zeo-Zn), and wastewater, IMI-treated wastewater and disinfected wastewater were compared. The numbers of residual FC on Swiss chard (Beta vulgaris) and in agricultural soil were determined over a 4-month period. The column using Zeo-Zn had a higher FC removal efficiency (2.98 log) than columns with other filter media and disinfection (1.87-2.57 log) due to the bactericidal properties of Zn(2+). The treatment of wastewater using Zeo-Zn and disinfection both decreased the accumulation of FC on plants and in soil to approximately 1-20 MPN/g dry matter. IMI-treated wastewater using the column with Zeo-Zn was suitable for unrestricted agricultural use, complied with Mexican regulations (as did disinfected wastewater) and had a low risk of FC contamination of plants and soil.

  3. Isolation and characterization of a thermotolerant ammonia-oxidizing bacterium Nitrosomonas sp. JPCCT2 from a thermal power station.

    Science.gov (United States)

    Itoh, Yoshikane; Sakagami, Keiko; Uchino, Yoshihito; Boonmak, Chanita; Oriyama, Tetsuro; Tojo, Fuyumi; Matsumoto, Mitsufumi; Morikawa, Masaaki

    2013-01-01

    A thermotolerant ammonia-oxidizing bacterium strain JPCCT2 was isolated from activated sludge in a thermal power station. Cells of JPCCT2 are short non-motile rods or ellipsoidal. Molecular phylogenetic analysis of 16S rRNA gene sequences demonstrated that JPCCT2 belongs to the genus Nitrosomonas with the highest similarity to Nitrosomonas nitrosa Nm90 (100%), Nitrosomonas sp. Nm148 (99.7%), and Nitrosomonas communis Nm2 (97.7%). However, G+C content of JPCCT2 DNA was 49.1 mol% and clearly different from N. nitrosa Nm90, 47.9%. JPCCT2 was capable of growing at temperatures up to 48 °C, while N. nitrosa Nm90 and N. communis Nm2 could not grow at 42°C. Moreover, JPCCT2 grew similarly at concentrations of carbonate 0 and 5 gL(-1). This is the first report that Nitrosomonas bacterium is capable of growing at temperatures higher than 37°C.

  4. Thermotolerant bacteriocin-producing lactic acid bacteria isolated from thai local fermented foods and their bacteriocin productivity.

    Science.gov (United States)

    Leelavatcharamas, Vichai; Arbsuwan, Nida; Apiraksakorn, Jirawan; Laopaiboon, Pattana; Kishida, Masao

    2011-03-01

    Twenty-one samples of Thai local fermented foods were screened for thermotolerant bacteriocin-producing lactic acid bacteria. From 529 isolates of lactic acid bacteria, 121 isolates were able to inhibit the growth of certain bacterial strains. Of these 121 isolates, only 11 produced antibacterial agents that were capable of inhibiting the growth of multiple bacterial strains in a liquid medium. One strain (KKU 170) of these 11 isolates produced an antibacterial agent that could strongly inhibit the growth of selected strains of gram-positive bacteria including Listeria sp. The antibacterial agent produced by the strain KKU 170 was identified as a bacteriocin since it was inactivated by proteinase K treatment. The strain KKU 170 was identified as Pediococcus acidilactici by both biochemical tests and molecular biological techniques. Optimal production of bacteriocin by the strain KKU 170 was found in culture medium containing 0.2% glucose, at an initial culture pH of 6.5, and temperature of 45 ºC. The maximum bacteriocin activity (1600 AU ml(-1)) was reached at the late exponential phase of growth and displayed primary metabolite production. The partially purified bacteriocin of the strain KKU 170 was tolerant to heat treatment at 121 ºC for 30 min.

  5. Identification of novel genes responsible for ethanol and/or thermotolerance by transposon mutagenesis in Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hyun-Soo [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Sciences; Kim, Na-Rae [Ewha Womans Univ., Seoul (Korea, Republic of). Div. of Life and Pharmaceutical Sciences; Yang, Jungwoo [Ewha Womans Univ., Seoul (Korea, Republic of). Microbial Resources Research Center; Choi, Wonja [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Sciences; Ewha Womans Univ., Seoul (Korea, Republic of). Div. of Life and Pharmaceutical Sciences; Ewha Womans Univ., Seoul (Korea, Republic of). Microbial Resources Research Center

    2011-08-15

    Saccharomyces cerevisiae strains tolerant to ethanol and heat stresses are important for industrial ethanol production. In this study, five strains (Tn 1-5) tolerant to up to 15% ethanol were isolated by screening a transposon-mediated mutant library. Two of them displayed tolerance to heat (42 C). The determination of transposon insertion sites and Northern blot analysis identified seven putative genes (CMP2, IMD4, SSK2, PPG1, DLD3, PAM1, and MSN2) and revealed simultaneous down-regulations of CMP2 and IMD4, and SSK2 and PPG1, down-regulation of DLD3, and disruptions of the open reading frame of PAM1 and MSN2, indicating that ethanol and/or heat tolerance can be conferred. Knockout mutants of these seven individual genes were ethanol tolerant and three of them (SSK2, PPG1, and PAM1) were tolerant to heat. Such tolerant phenotypes reverted to sensitive phenotypes by the autologous or overexpression of each gene. Five transposon mutants showed higher ethanol production and grew faster than the control strain when cultured in rich media containing 30% glucose and initial 6% ethanol at 30 C. Of those, two thermotolerant transposon mutants (Tn 2 and Tn 3) exhibited significantly enhanced growth and ethanol production compared to the control at 42 C. The genes identified in this study may provide a basis for the application in developing industrial yeast strains. (orig.)

  6. An evaluation of potential interferences in a fluorimetric assay for the rapid detection of thermotolerant coliforms in sewage.

    Science.gov (United States)

    Davies, C M; Apte, S C

    2000-02-01

    A 1-h fluorimetric assay of beta-D-galactosidase activity was evaluated for determining thermotolerant coliforms (TTC) in sewage samples. Above TTC concentrations of 2.3 x 103 colony-forming units (cfu) 100 ml-1, the assay response was related to TTC concentration. However, below this concentration, a large background signal was observed which was independent of TTC concentration. A separation scheme involving various filtration treatments and additions of a beta-D-galactosidase inhibitor was devised and used to quantify the sources of this anomalous assay response. The interferences encountered were largely due to the presence in sewage of non-specific cell-free enzymes or other cell-free substances that were capable of hydrolysing the fluorogenic substrate. Despite this apparent limitation, the fluorimetric enzyme assay has potential as an 'early warning' indicator of treatment process failure and gross sewage contamination and leakage in situations where TTC concentrations exceed 2.3 x 103 cfu 100 ml-1

  7. Artificial evolution by viability rather than competition.

    Directory of Open Access Journals (Sweden)

    Andrea Maesani

    Full Text Available Evolutionary algorithms are widespread heuristic methods inspired by natural evolution to solve difficult problems for which analytical approaches are not suitable. In many domains experimenters are not only interested in discovering optimal solutions, but also in finding the largest number of different solutions satisfying minimal requirements. However, the formulation of an effective performance measure describing these requirements, also known as fitness function, represents a major challenge. The difficulty of combining and weighting multiple problem objectives and constraints of possibly varying nature and scale into a single fitness function often leads to unsatisfactory solutions. Furthermore, selective reproduction of the fittest solutions, which is inspired by competition-based selection in nature, leads to loss of diversity within the evolving population and premature convergence of the algorithm, hindering the discovery of many different solutions. Here we present an alternative abstraction of artificial evolution, which does not require the formulation of a composite fitness function. Inspired from viability theory in dynamical systems, natural evolution and ethology, the proposed method puts emphasis on the elimination of individuals that do not meet a set of changing criteria, which are defined on the problem objectives and constraints. Experimental results show that the proposed method maintains higher diversity in the evolving population and generates more unique solutions when compared to classical competition-based evolutionary algorithms. Our findings suggest that incorporating viability principles into evolutionary algorithms can significantly improve the applicability and effectiveness of evolutionary methods to numerous complex problems of science and engineering, ranging from protein structure prediction to aircraft wing design.

  8. Cellular thermotolerance is independent of HSF 1 expression in zebu and crossbred non-lactating cattle

    Science.gov (United States)

    Gill, Jaspreet Kaur; Arora, J. S.; Sunil Kumar, B. V.; Mukhopadhyay, C. S.; Kaur, Simarjeet; Kashyap, Neeraj

    2017-09-01

    Heat stress is an important domain of research in livestock due to its negative impact on production and disease resistance. The augmentation of stress in the body stimulates the antioxidative activity comprising various enzymes (viz., catalase, superoxide dismutase), metabolites (reduced glutathione, etc.), vitamins, minerals, etc. to combat the situation. The major key players involved in regulation of heat shock response in eukaryotes are the transcription factors, called as heat shock factors (HSF). They activate the heat shock protein (HSP) genes by binding to their promoters. Lymphocytes are considered to be the best model to evaluate the immunity in any living body as it contains plethora of white blood cells (WBCs).In this study, the peripheral blood mononuclear cells (PBMC) obtained from non-lactating Sahiwal vis-à-vis crossbred (Holstein Friesian × Sahiwal) cattle with 75% or more exotic inheritance were subjected to heat shock at 39, 41, and 43 °C in three different incubators, in vitro. The cell count and viability test of pre and post heat stress of concerned PBMCs indicated that the crossbreeds are more prone to heat stress as compared to Sahiwal. The reverse transcription PCR (qRT-PCR) expression data revealed an increment in HSF1 expression at 41 °C which subsequently declined (non-significantly) at 43 °C in both breeds post 1 h heat shock. However, the association between the HSF 1 expression and antioxidative activity through correlation analysis was found to be non-significant ( P expression level on the activity of enzymes involved in oxidative stress in vitro in zebu and crossbred cattle.

  9. In vitro pollen germination and pollen viability in passion fruit (Passiflora spp.

    Directory of Open Access Journals (Sweden)

    Taliane Leila Soares

    2013-12-01

    Full Text Available The use of Passiflora species for ornamental purposes has been recently developed, but little is known about pollen viability and the potential for crossing different species. The objective of this study was to evaluate the pollen viability of six Passiflora species collected from different physiological stages of development through in vitro germination and histochemical analysis using dyes. The pollen was collected in three stages (pre-anthesis, anthesis and post-anthesis. Three compositions of culture medium were used to evaluate the in vitro germination, and two dyes (2,3,5-triphenyltetrazolium chloride, or TTC, and Lugol's solution were used for the histochemical analysis. The culture medium containing 0.03% Ca(NO3 4H2O, 0.02% of Mg(SO4 .7H2O, 0.01% of KNO3, 0,01% of H3BO3, 15% sucrose, and 0.8% agar, pH 7.0, showed a higher percentage of pollen grains germinated. Anthesis is the best time to collect pollen because it promotes high viability and germination. The Lugol's solution and TTC dye overestimated the viability of pollen, as all accessions showed high viability indices when compared with the results obtained in vitro.

  10. Thermodynamic study of residual heat from a high temperature nuclear reactor to analyze its viability in cogeneration processes; Estudio termodinamico del calor residual de un reactor nuclear de alta temperatura para analizar su viabilidad en procesos de cogeneracion

    Energy Technology Data Exchange (ETDEWEB)

    Santillan R, A.; Valle H, J.; Escalante, J. A., E-mail: santillanaura@gmail.com [Universidad Politecnica Metropolitana de Hidalgo, Boulevard acceso a Tolcayuca 1009, Ex-Hacienda San Javier, 43860 Tolcayuca, Hidalgo (Mexico)

    2015-09-15

    In this paper the thermodynamic study of a nuclear power plant of high temperature at gas turbine (GTHTR300) is presented for estimating the exploitable waste heat in a process of desalination of seawater. One of the most studied and viable sustainable energy for the production of electricity, without the emission of greenhouse gases, is the nuclear energy. The fourth generation nuclear power plants have greater advantages than those currently installed plants; these advantages have to do with security, increased efficiencies and feasibility to be coupled to electrical cogeneration processes. In this paper the thermodynamic study of a nuclear power plant type GTHTR300 is realized, which is selected by greater efficiencies and have optimal conditions for use in electrical cogeneration processes due to high operating temperatures, which are between 700 and 950 degrees Celsius. The aim of the study is to determine the heat losses and the work done at each stage of the system, determining where they are the greatest losses and analyzing in that processes can be taken advantage. Based on the study was appointed that most of the energy losses are in form of heat in the coolers and usually this is emitted into the atmosphere without being used. From the results a process of desalination of seawater as electrical cogeneration process is proposed. This paper contains a brief description of the operation of the nuclear power plant, focusing on operation conditions and thermodynamic characteristics for the implementation of electrical cogeneration process, a thermodynamic analysis based on mass and energy balance was developed. The results allow quantifying the losses of thermal energy and determining the optimal section for coupling of the reactor with the desalination process, seeking to have a great overall efficiency. (Author)

  11. Sperm viability staining in ecology and evolution: potential pitfalls

    DEFF Research Database (Denmark)

    Holman, Luke

    2009-01-01

    a number of interesting results, it has some potential pitfalls that have rarely been discussed. In the present paper, I review the major findings of ecology and evolution studies employing sperm viability staining and outline the method's principle limitations. The key problem is that the viability assay......The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining. Although sperm viability staining has produced...

  12. Assessing the Viability of Tiger Subpopulations in a Fragmented Landscape

    National Research Council Canada - National Science Library

    Matthew Linkie; Guillaume Chapron; Deborah J. Martyr; Jeremy Holden; Nigel Leader-Williams

    2006-01-01

    .... This study aimed to provide such information for tigers in the Kerinci Seblat (KS) region, Sumatra, by identifying and assessing subpopulation viability under different management strategies. 2...

  13. Fault Detection and Isolation using Viability Theory and Interval Observers

    Science.gov (United States)

    Ghaniee Zarch, Majid; Puig, Vicenç; Poshtan, Javad

    2017-01-01

    This paper proposes the use of interval observers and viability theory in fault detection and isolation (FDI). Viability theory develops mathematical and algorithmic methods for investigating the adaptation to viability constraints of evolutions governed by complex systems under uncertainty. These methods can be used for checking the consistency between observed and predicted behavior by using simple sets that approximate the exact set of possible behavior (in the parameter or state space). In this paper, fault detection is based on checking for an inconsistency between the measured and predicted behaviors using viability theory concepts and sets. Finally, an example is provided in order to show the usefulness of the proposed approach.

  14. The effects of storage conditions on the viability of ...

    African Journals Online (AJOL)

    The effects of storage conditions on the viability of enteropathogenics bacteria in biobanking of human stools: Cases of Yersinia enterocolitica, Salmonella enterica Typhimurium and Vibrio cholerae O: 1.

  15. Storage Viability and Optimization Web Service

    Energy Technology Data Exchange (ETDEWEB)

    Stadler, Michael; Marnay, Christ; Lai, Judy; Siddiqui, Afzal; Limpaitoon, Tanachai; Phan, Trucy; Megel, Olivier; Chang, Jessica; DeForest, Nicholas

    2010-10-11

    Non-residential sectors offer many promising applications for electrical storage (batteries) and photovoltaics (PVs). However, choosing and operating storage under complex tariff structures poses a daunting technical and economic problem that may discourage potential customers and result in lost carbon and economic savings. Equipment vendors are unlikely to provide adequate environmental analysis or unbiased economic results to potential clients, and are even less likely to completely describe the robustness of choices in the face of changing fuel prices and tariffs. Given these considerations, researchers at Lawrence Berkeley National Laboratory (LBNL) have designed the Storage Viability and Optimization Web Service (SVOW): a tool that helps building owners, operators and managers to decide if storage technologies and PVs merit deeper analysis. SVOW is an open access, web-based energy storage and PV analysis calculator, accessible by secure remote login. Upon first login, the user sees an overview of the parameters: load profile, tariff, technologies, and solar radiation location. Each parameter has a pull-down list of possible predefined inputs and users may upload their own as necessary. Since the non-residential sectors encompass a broad range of facilities with fundamentally different characteristics, the tool starts by asking the users to select a load profile from a limited cohort group of example facilities. The example facilities are categorized according to their North American Industry Classification System (NAICS) code. After the load profile selection, users select a predefined tariff or use the widget to create their own. The technologies and solar radiation menus operate in a similar fashion. After these four parameters have been inputted, the users have to select an optimization setting as well as an optimization objective. The analytic engine of SVOW is LBNL?s Distributed Energy Resources Customer Adoption Model (DER-CAM), which is a mixed

  16. Ponatinib reduces viability, migration, and functionality of human endothelial cells.

    Science.gov (United States)

    Gover-Proaktor, Ayala; Granot, Galit; Shapira, Saar; Raz, Oshrat; Pasvolsky, Oren; Nagler, Arnon; Lev, Dorit L; Inbal, Aida; Lubin, Ido; Raanani, Pia; Leader, Avi

    2017-06-01

    Tyrosine kinase inhibitors (TKIs) have revolutionized the prognosis of chronic myeloid leukemia. With the advent of highly efficacious therapy, the focus has shifted toward managing TKI adverse effects, such as vascular adverse events (VAEs). We used an in vitro angiogenesis model to investigate the TKI-associated VAEs. Our data show that imatinib, nilotinib, and ponatinib reduce human umbilical vein endothelial cells (HUVECs) viability. Pharmacological concentrations of ponatinib induced apoptosis, reduced migration, inhibited tube formation of HUVECs, and had a negative effect on endothelial progenitor cell (EPC) function. Furthermore, in HUVECs transfected with VEGF receptor 2 (VEGFR2), the effect of ponatinib on tube formation and on all parameters representing normal endothelial cell function was less prominent than in control cells. This is the first report regarding the pathogenesis of ponatinib-associated VAEs. The antiangiogenic effect of ponatinib, possibly mediated by VEGFR2 inhibition, as shown in our study, is another piece in the intricate puzzle of TKI-associated VAEs.

  17. Incorporating evolutionary processes into population viability models.

    Science.gov (United States)

    Pierson, Jennifer C; Beissinger, Steven R; Bragg, Jason G; Coates, David J; Oostermeijer, J Gerard B; Sunnucks, Paul; Schumaker, Nathan H; Trotter, Meredith V; Young, Andrew G

    2015-06-01

    We examined how ecological and evolutionary (eco-evo) processes in population dynamics could be better integrated into population viability analysis (PVA). Complementary advances in computation and population genomics can be combined into an eco-evo PVA to offer powerful new approaches to understand the influence of evolutionary processes on population persistence. We developed the mechanistic basis of an eco-evo PVA using individual-based models with individual-level genotype tracking and dynamic genotype-phenotype mapping to model emergent population-level effects, such as local adaptation and genetic rescue. We then outline how genomics can allow or improve parameter estimation for PVA models by providing genotypic information at large numbers of loci for neutral and functional genome regions. As climate change and other threatening processes increase in rate and scale, eco-evo PVAs will become essential research tools to evaluate the effects of adaptive potential, evolutionary rescue, and locally adapted traits on persistence. © 2014 Society for Conservation Biology.

  18. FT-IR Characterization of Pollen Biochemistry, Viability, and Germination Capacity in Saintpaulia H. Wendl. Genotypes

    Directory of Open Access Journals (Sweden)

    Erzsebet Buta

    2015-01-01

    Full Text Available FT-IR characterization of pollen biochemistry was analyzed to detect possible connection with the viability (by staining with potassium iodide, 25% and the germination capacity (on solid nutrient medium, in 15 Saintpaulia genotypes. Vibrational spectroscopy indicates that the pollen of S. ionantha genotype “Red Velvet” is rich in proteins, lipids, triglycerides, and esters and has a viability of 88.4% and a low germination capacity (27.16%. For S. ionantha “Jolly Red” and “Lucky Ladybug” genotypes, pollen showed high viability (88.81–91.49% and low germination capacity (23.02–9.17%, even though the pollen is rich in carbohydrates. S. ionantha “Aloha Orchid” genotype has the highest percentage of viability (94.32% and germination capacity (45.73% and a rich content of carbohydrates and polygalacturonic acids. In S. rupicola and S. ionantha genotypes, the rich content of polygalacturonic acids, lipids, and carbohydrates favourably influenced the germination capacity. Spectroscopic result indicates, through different absorbance band intensity, a possible link between biochemical composition, viability, and germination capacity of Saintpaulia pollen. To determine exactly the relation between biochemistry and biological processes, it is necessary to initiate quantitative researches.

  19. Effect of electrical charges and fields on injury and viability of airborne bacteria.

    Science.gov (United States)

    Mainelis, Gediminas; Górny, Rafał L; Reponen, Tiina; Trunov, Mikhaylo; Grinshpun, Sergey A; Baron, Paul; Yadav, Jagjit; Willeke, Klaus

    2002-07-20

    In this study, the effects of the electric charges and fields on the viability of airborne microorganisms were investigated. The electric charges of different magnitude and polarity were imparted on airborne microbial cells by a means of induction charging. The airborne microorganisms carrying different electric charge levels were then extracted by an electric mobility analyzer and collected using a microbial sampler. It was found that the viability of Pseudomonas fluorescens bacteria, used as a model for sensitive bacteria, carrying a net charge from 4100 negative to 30 positive elementary charges ranged between 40% and 60%; the viability of the cells carrying >2700 positive charges was below 1.5%. In contrast, the viability of the stress-resistant spores of Bacillus subtilis var. niger (used as simulant of anthrax-causing Bacillus anthracis spores when testing bioaerosol sensors in various studies), was not affected by the amount of electric charges on the spores. Because bacterial cells depend on their membrane potential for basic metabolic activities, drastic changes occurring in the membrane potential during aerosolization and the local electric fields induced by the imposed charges appeared to affect the sensitive cells' viability. These findings facilitate applications of electric charging for environmental control purposes involving sterilization of bacterial cells by imposing high electric charges on them. The findings from this study can also be used in the development of new bioaerosol sampling methods based on electrostatic principles. Copyright 2002 Wiley Periodicals, Inc.

  20. Advances and Challenges in Viability Detection of Foodborne Pathogens

    Directory of Open Access Journals (Sweden)

    Dexin Zeng

    2016-11-01

    Full Text Available Foodborne outbreaks are a serious public health and food safety concern worldwide. There is a great demand for rapid, sensitive, specific, and accurate methods to detect microbial pathogens in foods. Conventional methods based on cultivation of pathogens have been the gold standard protocols; however, they take up to a week to complete. Molecular assays such as polymerase chain reaction (PCR, sequencing, microarray technologies have been widely used in detection of foodborne pathogens. Among molecular assays, PCR technology conventional and real-time PCR (qPCR is most commonly used in the foodborne pathogen detection because of its high sensitivity and specificity. However, a major drawback of PCR is its inability to differentiate the DNA from dead and viable cells, and this is a critical factor for the food industry, regulatory agencies and the consumer. To remedy this shortcoming, researchers have used biological dyes such as ethidium monoazide (EMA and propidium monoazide (PMA to pretreat samples before DNA extraction to intercalate the DNA of dead cells in food samples, and then proceed with regular DNA preparation and qPCR. By combining PMA treatment with qPCR (PMA-qPCR, scientists have applied this technology to detect viable cells of various bacterial pathogens in foods. The incorporation of PMA into PCR-based assays for viability detection of pathogens in foods has increased significantly in the last decade. On the other hand, some downsides with this approach have been noted, particularly to achieve complete suppression of signal of DNA from the dead cells present in some particular food matrix. Nowadays, there is a tendency of more and more researchers adapting this approach for viability detection; and a few commercial kits based on PMA are available in the market. As time goes on, more scientists apply this approach to a broader range of pathogen detections, this viability approach (PMA or other chemicals such as platinum compound

  1. Purification, characterization, and overexpression of an endo-1,4-β-mannanase from thermotolerant Bacillus sp. SWU60.

    Science.gov (United States)

    Seesom, Weeranuch; Thongket, Polphet; Yamamoto, Tomohiro; Takenaka, Shigeo; Sakamoto, Tatsuji; Sukhumsirichart, Wasana

    2017-03-01

    Endo-β-1,4-mannanases are important catalytic agents in several industries. The enzymes randomly cleave the β-1,4-linkage in the mannan backbone and release short β-1,4-mannooligosaccharides and mannose. In the present study, mannanase (ManS2) from thermotolerant Bacillus sp. SWU60 was purified, characterized, and its gene was cloned and overexpressed in Escherichia coli. ManS2 was purified from culture filtrate (300 ml) by using hydrophobic, ion-exchange, and size-exclusive liquid chromatography. The apparent molecular mass was 38 kDa. Optimal pH and temperature for enzyme activity were 6.0 and 60 °C, respectively. The enzyme was stable up to 60 °C for 1 h and at pH 5-9 at 4 °C for 16 h. Its enzyme activity was inhibited by Hg(2+). The full-length mans2 gene was 1,008 bp, encoding a protein of 336 amino acids. Amino acid sequence analysis revealed that it belonged to glycoside hydrolase family 26. Konjac glucomannan was a favorable substrate for recombinant ManS2 (rManS2). rManS2 also degraded galactomannan from locust bean gum, indicating its potential for production of glucomanno- and galactomanno-oligosaccharides. Both native and recombinant ManS2 from Bacillus sp. SWU60 can be applied in several industries especially food and feed.

  2. Effect of Diluent and Relative Humidity on Apparent Viability of Airborne Pasteurella pestis

    Science.gov (United States)

    Won, William D.; Ross, Harold

    1966-01-01

    Airborne Pasteurella pestis (A-1122) at low humidities [20 to 50% relative humidity (RH)] exhibited exponential decay when either 1% peptone or Heart Infusion Broth (HIB) was used as the diluent in the viable assay system. At higher RH values (65 and 87%), however, the 1% peptone diluent adversely affected the viability assay. In contrast, HIB as diluent was remarkably effective in demonstrating a higher number of viable cells in aerosols held at high RH values. Similarly, with HIB as diluent, aerosols were shown to contain viable cells during 90 min of observation; with 1% peptone, viability was not detectable after 20 min in the airborne state. PMID:5970462

  3. Effect of diluent and relative humidity on apparent viability of airborne Pasteurella pestis.

    Science.gov (United States)

    Won, W D; Ross, H

    1966-09-01

    Airborne Pasteurella pestis (A-1122) at low humidities [20 to 50% relative humidity (RH)] exhibited exponential decay when either 1% peptone or Heart Infusion Broth (HIB) was used as the diluent in the viable assay system. At higher RH values (65 and 87%), however, the 1% peptone diluent adversely affected the viability assay. In contrast, HIB as diluent was remarkably effective in demonstrating a higher number of viable cells in aerosols held at high RH values. Similarly, with HIB as diluent, aerosols were shown to contain viable cells during 90 min of observation; with 1% peptone, viability was not detectable after 20 min in the airborne state.

  4. Efficacy of propidium iodide and FUN-1 stains for assessing viability in basidiospores of Rhizopogon roseolus.

    Science.gov (United States)

    Fernández-Miranda, Elena; Majada, Juan; Casares, Abelardo

    2017-01-01

    The use of spores in applications of ectomycorrhizal fungi requires information regarding spore viability and germination, especially in genera such as Rhizopogon with high rates of spore dormancy. The authors developed a protocol to assess spore viability of Rhizopogon roseolus using four vital stains to quantify spore viability and germination and to optimize storage procedures. They showed that propidium iodide is an excellent stain for quantifying nonviable spores. Observing red fluorescent intravacuolar structures following staining with 2-chloro-4-(2,3-dihydro-3-methyl-(benzo-1,3-thiazol-2-yl)-methylidene)-1-phenylquinolinium iodide (FUN-1) can help identify viable spores that are activated. At 6 mo and 1 y, the spores kept in a water suspension survived better than those left within intact, dry gasterocarps. Our work highlights the importance of temperature, nutrients, and vitamins for maturation and germination of spores of R. roseolus during 1 y of storage.

  5. Influence of electrospun scaffolds prepared from distinct polymers on proliferation and viability of endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Matveeva, V. G., E-mail: matveeva-vg@mail.ru; Antonova, L. V., E-mail: antonova.la@mail.ru; Velikanova, E. A.; Sergeeva, E. A.; Krivkina, E. O.; Glushkova, T. V.; Kudryavtseva, Yu. A.; Barbarash, O. L.; Barbarash, L. S. [Research Institute for Complex Issues of Cardiovascular Diseases, Kemerovo, 650002 (Russian Federation)

    2015-10-27

    We compared electrospun nonwoven scaffolds from polylactic acid (PLA), polycaprolactone (PCL), and polyhydroxybutyrate/valerate (PHBV)/polycaprolactone (PHBV/PCL). The surface of PHBV/PCL and PCL scaffolds was highly porous and consisted of randomly distributed fibers, whilst the surface of PLA scaffolds consisted of thin straight fibers, which located more sparsely, forming large pores. Culture of EA.hy 926 endothelial cells on these scaffolds during 7 days and further fluorescent microscopy demonstrated that the surface of PHBV/PCL scaffolds was most favorable for efficient adhesion, proliferation, and viability of endothelial cells. The lowest proliferation rate and cell viability were detected on PLA scaffolds. Therefore, PHBV/PCL electrospun nonwoven scaffolds demonstrated the best results regarding endothelial cell proliferation and viability as compared to PCL and PLA scaffolds.

  6. Colony Failure Linked to Low Sperm Viability in Honey Bee (Apis mellifera Queens and an Exploration of Potential Causative Factors.

    Directory of Open Access Journals (Sweden)

    Jeffery S Pettis

    Full Text Available Queen health is closely linked to colony performance in honey bees as a single queen is normally responsible for all egg laying and brood production within the colony. In the U. S. in recent years, queens have been failing at a high rate; with 50% or greater of queens replaced in colonies within 6 months when historically a queen might live one to two years. This high rate of queen failure coincides with the high mortality rates of colonies in the US, some years with >50% of colonies dying. In the current study, surveys of sperm viability in US queens were made to determine if sperm viability plays a role in queen or colony failure. Wide variation was observed in sperm viability from four sets of queens removed from colonies that beekeepers rated as in good health (n = 12; average viability = 92%, were replacing as part of normal management (n = 28; 57%, or where rated as failing (n = 18 and 19; 54% and 55%. Two additional paired set of queens showed a statistically significant difference in viability between colonies rated by the beekeeper as failing or in good health from the same apiaries. Queens removed from colonies rated in good health averaged high viability (ca. 85% while those rated as failing or in poor health had significantly lower viability (ca. 50%. Thus low sperm viability was indicative of, or linked to, colony performance. To explore the source of low sperm viability, six commercial queen breeders were surveyed and wide variation in viability (range 60-90% was documented between breeders. This variability could originate from the drones the queens mate with or temperature extremes that queens are exposed to during shipment. The role of shipping temperature as a possible explanation for low sperm viability was explored. We documented that during shipment queens are exposed to temperature spikes ( 40°C and these spikes can kill 50% or more of the sperm stored in queen spermathecae in live queens. Clearly low sperm viability is

  7. Colony Failure Linked to Low Sperm Viability in Honey Bee (Apis mellifera) Queens and an Exploration of Potential Causative Factors.

    Science.gov (United States)

    Pettis, Jeffery S; Rice, Nathan; Joselow, Katie; vanEngelsdorp, Dennis; Chaimanee, Veeranan

    2016-01-01

    Queen health is closely linked to colony performance in honey bees as a single queen is normally responsible for all egg laying and brood production within the colony. In the U. S. in recent years, queens have been failing at a high rate; with 50% or greater of queens replaced in colonies within 6 months when historically a queen might live one to two years. This high rate of queen failure coincides with the high mortality rates of colonies in the US, some years with >50% of colonies dying. In the current study, surveys of sperm viability in US queens were made to determine if sperm viability plays a role in queen or colony failure. Wide variation was observed in sperm viability from four sets of queens removed from colonies that beekeepers rated as in good health (n = 12; average viability = 92%), were replacing as part of normal management (n = 28; 57%), or where rated as failing (n = 18 and 19; 54% and 55%). Two additional paired set of queens showed a statistically significant difference in viability between colonies rated by the beekeeper as failing or in good health from the same apiaries. Queens removed from colonies rated in good health averaged high viability (ca. 85%) while those rated as failing or in poor health had significantly lower viability (ca. 50%). Thus low sperm viability was indicative of, or linked to, colony performance. To explore the source of low sperm viability, six commercial queen breeders were surveyed and wide variation in viability (range 60-90%) was documented between breeders. This variability could originate from the drones the queens mate with or temperature extremes that queens are exposed to during shipment. The role of shipping temperature as a possible explanation for low sperm viability was explored. We documented that during shipment queens are exposed to temperature spikes ( 40°C) and these spikes can kill 50% or more of the sperm stored in queen spermathecae in live queens. Clearly low sperm viability is linked to

  8. Establishing guidelines to retain viability of probiotics during spray drying

    NARCIS (Netherlands)

    Perdana, J.A.; Fox, M.B.; Boom, R.M.; Schutyser, M.A.I.

    2014-01-01

    We present a model-based approach to map processing conditions suitable to spray dry probiotics with minimal viability loss. The approach combines the drying history and bacterial inactivation kinetics to predict the retention of viability after drying. The approach was used to systematically assess

  9. Evaluation of pollen viability, stigma receptivity and fertilization ...

    African Journals Online (AJOL)

    To provide theoretical basis for artificial pollination in Lagerstroemia indica L., pollen viability and stigma receptivity were tested and the morphological change of stigma was observed. Pollen viability tested by in vitro culture, stigma receptivity examined by benzidine-H2O2 testing and fruit set estimated by field artificial ...

  10. Viability of dielectrophoretically trapped neuronal cortical cells in culture

    NARCIS (Netherlands)

    Heida, Tjitske; Vulto, P; Rutten, Wim; Marani, Enrico

    2001-01-01

    Negative dielectrophoretic trapping of neural cells is an efficient way to position neural cells on the electrode sites of planar micro-electrode arrays. The preservation of viability of the neural cells is essential for this approach. This study investigates the viability of postnatal cortical rat

  11. 37 CFR 1.807 - Viability of deposit.

    Science.gov (United States)

    2010-07-01

    ... 37 Patents, Trademarks, and Copyrights 1 2010-07-01 2010-07-01 false Viability of deposit. 1.807... COMMERCE GENERAL RULES OF PRACTICE IN PATENT CASES Biotechnology Invention Disclosures Deposit of Biological Material § 1.807 Viability of deposit. (a) A deposit of biological material that is capable of...

  12. Evaluation of pollen viability, stigma receptivity and fertilization ...

    African Journals Online (AJOL)

    AJL

    2013-11-13

    Nov 13, 2013 ... To provide theoretical basis for artificial pollination in Lagerstroemia indica L., pollen viability and stigma receptivity were tested and the morphological change of stigma was observed. Pollen viability tested by in vitro culture, stigma receptivity examined by benzidine-H2O2 testing and fruit set estimated.

  13. The Economy and Democracy: Viability and Challenges for ...

    African Journals Online (AJOL)

    The Economy and Democracy: Viability and Challenges for Sustainable Democratisation in Nigeria. ... Economic and Policy Review ... the viability for developing sustainable democracy in Nigeria against the background of the country's enormous economic potentials and the economic reforms introduced following the ...

  14. Pollen viability and germination in Jatropha ribifolia and Jatropha ...

    African Journals Online (AJOL)

    The aim of this work is to assess pollen viability using the staining technique and in vitro germination with different concentrations of sucrose in Jatropha ribifolia and Jatropha mollissima, contributing to the knowledge of the reproductive biology and subsidizing their conservation, management and utilization. Pollen viability ...

  15. Viability, Advantages and Design Methodologies of M-Learning Delivery

    Science.gov (United States)

    Zabel, Todd W.

    2010-01-01

    The purpose of this study was to examine the viability and principle design methodologies of Mobile Learning models in developing regions. Demographic and market studies were utilized to determine the viability of M-Learning delivery as well as best uses for such technologies and methods given socioeconomic and political conditions within the…

  16. Economic viability of transmission capacity expansion at high wind penetrations

    DEFF Research Database (Denmark)

    Østergaard, Poul Alberg

    2005-01-01

    With growing wind power penetrations in many countries, grid and system integration becomes more and more important issues. This is particularly the case in countries or regions with good wind resources as well as substantial installed wind power capacity as found in e.g. Northern Europe. At 20......% penetration in Western Denmark, the issue is pertinent here in relation to future plans of further expansion which is planned in accordance with the Danish Government’s climate change mitigation initiatives. This paper analyses the potential economic benefit of selling excess electricity production...... investments and analyses of the Nord Pool price variations. The analyses are done for varying degrees of wind power penetrations ranging from 20% of the West Danish electricity demand up to 100% of the demand. The analyses demonstrate, that while there is an economic potential for some expansion in some years...

  17. Assessment of cryopreserved donor skin viability: the experience of the regional tissue bank of Siena.

    Science.gov (United States)

    Pianigiani, E; Tognetti, L; Ierardi, F; Mariotti, G; Rubegni, P; Cevenini, G; Perotti, R; Fimiani, M

    2016-06-01

    Skin allografts from cadaver donors are an important resource for treating extensive burns, slow-healing wounds and chronic ulcers. A high level of cell viability of cryopreserved allografts is often required, especially in burn surgery, in Italy. Thus, we aimed to determine which conditions enable procurement of highly viable skin in our Regional Skin Bank of Siena. For this purpose, we assessed cell viability of cryopreserved skin allografts procured between 2011 and 2013 from 127 consecutive skin donors, before and after freezing (at day 15, 180, and 365). For each skin donor, we collected data concerning clinical history (age, sex, smoking, phototype, dyslipidemia, diabetes, cause of death), donation process (multi-tissue or multi-organ) and timing of skin procurement (assessment of intervals such as death-harvesting, harvesting-banking, death-banking). All these variables were analysed in the whole case study (127 donors) and in different groups (e.g. multi-organ donors, non refrigerated multi-tissue donors, refrigerated multi-tissue donors) for correlations with cell viability. Our results indicated that cryopreserved skin allografts with higher cell viability were obtained from female, non smoker, heartbeating donors died of cerebral haemorrhage, and were harvested within 2 h of aortic clamping and banked within 12 h of harvesting (13-14 h from clamping). Age, cause of death and dyslipidaemia or diabetes did not appear to influence cell viability. To maintain acceptable cell viability, our skin bank needs to reduce the time interval between harvesting and banking, especially for refrigerated donors.

  18. Seed viability of five wild Saudi Arabian species by germination and X-ray tests

    Directory of Open Access Journals (Sweden)

    B.A. Al-Hammad

    2017-09-01

    Full Text Available Our objective was to evaluate the usefulness of the germination vs. the X-ray test in determining the initial viability of seeds of five wild species (Moringa peregrina, Abrus precatorius, Arthrocnemum macrostachyum, Acacia ehrenbergiana and Acacia tortilis from Saudi Arabia. Usually several days were required to determine the viability of all five species via germination tests. However, X-ray test will give immediate results on filled/viable seeds. Seeds of all species, except Acacia ehrenbergiana and Acacia tortilis showed high viability in both germination (96–72% at 25/15 °C, 94–70% at 35/25 °C and X-ray (100–80% test. Furthermore, there was a general agreement between the germination (19%, 14% at 25/15 °C and 17% and 12% at 35/25 °C and X-ray (8%, 4% tests in which seed viability of Acacia ehrenbergiana and Acacia tortilis was very low due to insect damaged embryo as shown in X-ray analysis. Seeds of Abruspreca torius have physical dormancy, which was broken by scarification in concentrated sulfuric acid (10 min, and they exhibited high viability in both the germination (83% at 25/15 °C and 81% at 35/25 °C and X-ray (96% tests. Most of the nongerminated seeds of the five species except those of Acacia ehrenbergiana and Acacia tortilis, were alive as judged by the tetrazolium test (TZ. Thus, for the five species examined, the X-ray test was proved to be a good and rapid predictor of seed viability.

  19. Development and viability of a translocated beaver Castor fiber population in the Netherlands

    NARCIS (Netherlands)

    Nolet, B.A.; Baveco, J.M.

    1996-01-01

    We monitored survival, reproduction and emigration of a translocated beaver Castor fiber population in the Netherlands for five years and used a stochastic model to assess its viability. Between 1988 and 1991, 42 beavers were released in the Biesbosch National Park. The mortality was initially high

  20. Viability and functional integrity of washed platelets

    Energy Technology Data Exchange (ETDEWEB)

    Pineda, A.A.; Zylstra, V.W.; Clare, D.E.; Dewanjee, M.K.; Forstrom, L.A.

    1989-07-01

    The viability and functional integrity of saline- and ACD-saline-washed platelets were compared with those of unwashed platelets. After template bleeding time (TBT) was measured, 15 healthy volunteers underwent plateletpheresis and ingested 600 mg of aspirin. Autologous /sup 111/In-labeled platelets were transfused: unwashed (n = 5), washed with 0.9 percent saline solution (SS) (n = 5), and washed with a buffered 12.6 percent solution of ACD-A in 0.9 percent saline solution (n = 5). After transfusion, we measured TBT at 1, 4, and 24 hours; platelet survival at 10 minutes and 1, 4, and 24 hours and daily for 6 days; and the percentage of uptake in liver and spleen by quantitative whole-body radionuclide scintigraphy at 24 and 190 hours. We found that saline washing affected platelet recovery, 23.47 +/- 12 percent (p less than 0.001) as compared to 52.43 +/- 17 percent (p less than 0.002) for ACD-saline and 73.17 +/- 8 percent for control; that saline washing resulted in a greater liver uptake than control and ACD-saline-washed platelets (31.9 +/- 8% (p less than 0.001) vs 17.7 +/- 4.1 and 19.3 +/- 2.1% (p greater than 0.1), respectively); that, unlike control and ACD-saline-washed platelets, saline-washed platelets did not shorten bleeding time; and that neither type of washing affected survival. Although ACD-saline washing affects recovery, it also results in intact function, normal survival, higher recovery than SS platelets, and no significant liver uptake.

  1. High Level Ethanol from Sugar Cane Molasses by a New Thermotolerant Saccharomyces cerevisiae Strain in Industrial Scale

    OpenAIRE

    Fadel, M.; Keera, Abeer A.; Mouafi, Foukia E.; Kahil, Tarek

    2013-01-01

    A new local strain of S. cerevisiae F-514, for ethanol production during hot summer season, using Egyptian sugar cane molasses was applied in Egyptian distillery factory. The inouluum was propagated through 300?L, 3?m3, and 12?m3 fermenters charged with diluted sugar cane molasses containing 4%-5% sugars. The yeast was applied in fermentation vessels 65?m3 working volume to study the varying concentrations of urea, DAP, orthophosphoric acid (OPA), and its combinations as well as magnesium sul...

  2. High Level Ethanol from Sugar Cane Molasses by a New Thermotolerant Saccharomyces cerevisiae Strain in Industrial Scale.

    Science.gov (United States)

    Fadel, M; Keera, Abeer A; Mouafi, Foukia E; Kahil, Tarek

    2013-01-01

    A new local strain of S. cerevisiae F-514, for ethanol production during hot summer season, using Egyptian sugar cane molasses was applied in Egyptian distillery factory. The inouluum was propagated through 300 L, 3 m(3), and 12 m(3) fermenters charged with diluted sugar cane molasses containing 4%-5% sugars. The yeast was applied in fermentation vessels 65 m(3) working volume to study the varying concentrations of urea, DAP, orthophosphoric acid (OPA), and its combinations as well as magnesium sulfate and inoculum size. The fermenter was allowed to stay for a period of 20 hours to give time for maximum conversion of sugars into ethanol. S. cerevisiae F-514 at molasses sugar level of 18% (w/v), inoculum size of 20% (v/v) cell concentration of 3.0 × 10(8)/mL, and combinations of urea, diammonium phosphate (DAP), orthophosphoric acid (OPA), and magnesium sulfate at amounts of 20, 10, 5, and 10 kg/65 m(3) working volume fermenters, respectively, supported maximum ethanol production (9.8%, v/v), fermentation efficiency (FE) 88.1%, and remaining sugars (RS) 1.22%. The fermentation resulted 13.4 g dry yeast/L contained 34.6% crude protein and 8.2% ash. By selecting higher ethanol yielding yeast strain and optimizing, the fermentation parameters both yield and economics of the fermentation process can be improved.

  3. Quantifying spore viability of the honey bee pathogen Nosema apis using flow cytometry.

    Science.gov (United States)

    Peng, Yan; Lee-Pullen, Tracey F; Heel, Kathy; Millar, A Harvey; Baer, Boris

    2014-05-01

    Honey bees are hosts to more than 80 different parasites, some of them being highly virulent and responsible for substantial losses in managed honey bee populations. The study of honey bee pathogens and their interactions with the bees' immune system has therefore become a research area of major interest. Here we developed a fast, accurate and reliable method to quantify the viability of spores of the honey bee gut parasite Nosema apis. To verify this method, a dilution series with 0, 25, 50, 75, and 100% live N. apis was made and SYTO 16 and Propidium Iodide (n = 35) were used to distinguish dead from live spores. The viability of spores in each sample was determined by flow cytometry and compared with the current method based on fluorescence microscopy. Results show that N. apis viability counts using flow cytometry produced very similar results when compared with fluorescence microscopy. However, we found that fluorescence microscopy underestimates N. apis viability in samples with higher percentages of viable spores, the latter typically being what is found in biological samples. A series of experiments were conducted to confirm that flow cytometry allows the use of additional fluorescent dyes such as SYBR 14 and SYTOX Red (used in combination with SYTO 16 or Propidium Iodide) to distinguish dead from live spores. We also show that spore viability quantification with flow cytometry can be undertaken using substantially lower dye concentrations than fluorescence microscopy. In conclusion, our data show flow cytometry to be a fast, reliable method to quantify N. apis spore viabilities, which has a number of advantages compared with existing methods. © 2013 International Society for Advancement of Cytometry.

  4. Shape evolution with temperature of a thermotolerant protein (PeaT1) in solution detected by small angle X-ray scattering.

    Science.gov (United States)

    Xing, Xueqing; Liu, Quan; Wang, Wei; Zhang, Kunhao; Li, Tang; Cai, Quan; Mo, Guang; Cheng, Weidong; Wang, Dehong; Gong, Yu; Chen, Zhongjun; Qiu, Dewen; Wu, Zhonghua

    2013-01-01

    The protein elicitor from Alternaria tenuissima (PeaT1) presented excellent thermotolerance and potential application in agriculture as a pesticide. Previous synchrotron radiation circular dichroism study demonstrated that the secondary structures in PeaT1 protein are reversible with temperature change. To further clarify the mechanism of its thermotolerance, synchrotron radiation small angle x-ray scattering (SAXS) technique was used to study the shape change of PeaT1 protein with temperature in this article. Ab initio structure restorations based on the SAXS data revealed that PeaT1 protein has a prolate shape with a P₂ symmetry axis along the prolate anisometric direction. With temperature increase, a gooseneck vase-like (25°C), to jug-like (55°C), then to oval (85°C) shape change can be found, and these shape changes are also approximately reversible with temperature decrease. PeaT1 protein contains two homogenous molecules, and each of them consists of F, NAC, T, and UBA domains. The structures of the four domains were predicted. Simulated annealing algorithm was used to superimpose the domain structures onto the SAXS shapes. It was found that all the structural domains have position rotation and translation with temperature change, but the NAC domains are relatively stable, playing a role of frame. This shape change information provides clues for further exploring its biological function and application. Copyright © 2012 Wiley Periodicals, Inc.

  5. The methionine-rich low-molecular-weight chloroplast heat-shock protein: evolutionary conservation and accumulation in relation to thermotolerance.

    Science.gov (United States)

    Downs, C; Heckathorn, S; Bryan, J; Coleman, J

    1998-02-01

    The evolutionary conservation of the low-molecular-weight chloroplast-localized heat-shock protein (LMW chlpHsp) in vascular plants was examined using immunological methods. An antibody (Abmet) specific to the LMW chlpHsp was produced using a synthetic 28-residue peptide containing the most conserved elements of its unique "methionine-rich domain" as an antigen. This antibody detected a heat-inducible low-molecular-weight chloroplast protein in plants of six divergent Anthophyta species, including C3, C4, CAM, monocot, and dicot species. Abmet also detected a LMW chlpHsp in species from the Divisions Psilotophyta, Equisetophyta, Polypodiophyta, and Ginkgophyta. A preliminary examination of the relationship between accumulation of the LMW chlpHsp and habitat was also conducted. Seven Anthophyta species originating from both warm- and cool-temperature habitats were grown at 28C and then heat stressed at 40C. A positive qualitative relationship between the accumulation of the LMW chlpHsp and organismal thermotolerance in these species was observed; similar results were obtained separately with four nonAnthophyta species. The strong evolutionary conservation of this LMW Hsp and its localization to the chloroplast, and the correlation between production of this protein and plant thermotolerance, suggest that the LMW chlpHsp plays an important role in adaptation to heat stress.

  6. Comparative genome analysis of a thermotolerant Escherichia coli obtained by Genome Replication Engineering Assisted Continuous Evolution (GREACE) and its parent strain provides new understanding of microbial heat tolerance.

    Science.gov (United States)

    Luan, Guodong; Bao, Guanhui; Lin, Zhao; Li, Yang; Chen, Zugen; Li, Yin; Cai, Zhen

    2015-12-25

    Heat tolerance of microbes is of great importance for efficient biorefinery and bioconversion. However, engineering and understanding of microbial heat tolerance are difficult and insufficient because it is a complex physiological trait which probably correlates with all gene functions, genetic regulations, and cellular metabolisms and activities. In this work, a novel strain engineering approach named Genome Replication Engineering Assisted Continuous Evolution (GREACE) was employed to improve the heat tolerance of Escherichia coli. When the E. coli strain carrying a mutator was cultivated under gradually increasing temperature, genome-wide mutations were continuously generated during genome replication and the mutated strains with improved thermotolerance were autonomously selected. A thermotolerant strain HR50 capable of growing at 50°C on LB agar plate was obtained within two months, demonstrating the efficiency of GREACE in improving such a complex physiological trait. To understand the improved heat tolerance, genomes of HR50 and its wildtype strain DH5α were sequenced. Evenly distributed 361 mutations covering all mutation types were found in HR50. Closed material transportations, loose genome conformation, and possibly altered cell wall structure and transcription pattern were the main differences of HR50 compared with DH5α, which were speculated to be responsible for the improved heat tolerance. This work not only expanding our understanding of microbial heat tolerance, but also emphasizing that the in vivo continuous genome mutagenesis method, GREACE, is efficient in improving microbial complex physiological trait. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Comparison and Recovery of Escherichia coli and Thermotolerant Coliforms in Water with a Chromogenic Medium Incubated at 41 and 44.5°C

    Science.gov (United States)

    Alonso, Jose L.; Soriano, Adela; Carbajo, Oscar; Amoros, Inmaculada; Garelick, Hemda

    1999-01-01

    This study compared the performance of a commercial chromogenic medium, CHROMagarECC (CECC), and CECC supplemented with sodium pyruvate (CECCP) with the membrane filtration lauryl sulfate-based medium (mLSA) for enumeration of Escherichia coli and non-E. coli thermotolerant coliforms (KEC). To establish that we could recover the maximum KEC and E. coli population, we compared two incubation temperature regimens, 41 and 44.5°C. Statistical analysis by the Fisher test of data did not demonstrate any statistically significant differences (P = 0.05) in the enumeration of E. coli for the different media (CECC and CECCP) and incubation temperatures. Variance analysis of data performed on KEC counts showed significant differences (P = 0.01) between KEC counts at 41 and 44.5°C on both CECC and CECCP. Analysis of variance demonstrated statistically significant differences (P = 0.05) in the enumeration of total thermotolerant coliforms (TTCs) on CECC and CECCP compared with mLSA. Target colonies were confirmed to be E. coli at a rate of 91.5% and KEC of likely fecal origin at a rate of 77.4% when using CECCP incubated at 41°C. The results of this study showed that CECCP agar incubated at 41°C is efficient for the simultaneous enumeration of E. coli and KEC from river and marine waters. PMID:10427079

  8. Comparative evaluation of chromogenic agar CM1046 and mFC agar for detection of E. coli and thermotolerant coliform bacteria from water samples.

    Science.gov (United States)

    Wohlsen, T D

    2011-08-01

    The equivalence of Oxoid (CM 1046) Brilliance((TM)) E. coli/coliform selective agar to mFC agar, as used in the Australian/New Zealand Standard Method to detect thermotolerant coliforms and Escherichia coli in water samples, was assessed. A total of 244 water samples were analysed in parallel over a 5-month period. Sewage effluent samples (n = 131, sites = 43), freshwater (n = 62, sites = 18) and marine/brackish water samples (n = 51, sites = 23) were analysed. The Wilcoxon matched-pairs signed-ranks test showed a varying degree of statistical difference between the two methods. All matrices had a higher recovery in the trial method. Enterococci faecalis, Aeromonas spp. and Vibrio spp. did not grow on the CM1046 agar, and Pseudomonas aeruginosa and Enterobacter aerogenes were inhibited. The use of CM 1046 for the detection and enumeration of E. coli and thermotolerant coliforms in water samples is a suitable alternative to the AS/NZS Standard Method. The use of CM1046 agar was less labour intensive and time consuming, as no secondary confirmation steps were required. Confirmed results could be reported within 24 h of sample analysis, as compared to 48 h with the reference method. Public health concerns can be addressed in a more efficient manner. © 2011 Unitywater. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

  9. Prevalence of Thermotolerant Campylobacter spp. in Chicken Meat in Croatia and Multilocus Sequence Typing of a Small Subset of Campylobacter jejuni and Campylobacter coli Isolates

    Directory of Open Access Journals (Sweden)

    Andrea Humski

    2016-01-01

    Full Text Available In order to detect thermotolerant Campylobacter spp., 241 samples of fresh chicken meat, at retail in Croatia, were analysed according to a standard method, followed by biochemical test and molecular polymerase chain reaction/restriction enzyme analysis for exact species determination. Campylobacter spp. prevalence was 73.86 %. Campylobacter jejuni and Campylobacter coli were isolated from 53.53 and 15.35 % of the samples, respectively. In 4.98 % of isolates thermotolerant Campylobacter spp. were not determined. The multi locus sequence typing method was used to evaluate genetic diversity of eight Campylobacter jejuni and four Campylobacter coli isolates. To our knowledge, these results of genotyping provided the first data on the presence of sequence types (STs and clonal complexes (CCs of Campylobacter jejuni and C. coli isolates in Croatia. By applying the multilocus sequence typing, a new allele of tkt gene locus was discovered and marked tkt508. The C. jejuni ST 6182 and C. coli ST 6183 genotypes were described for the fi rst time, and all other identified genotypes were clustered in the previously described sequence types and clonal complexes. These findings provide useful information on the prevalence and epidemiology of Campylobacter jejuni and C. coli in Croatia.

  10. Effect of thermo-tolerant yeast on intake and nutrient digestibility's in Murrah buffalo steers (Bubalus bubalis fed straw based complete diet

    Directory of Open Access Journals (Sweden)

    B. Bhima

    2014-07-01

    Full Text Available Aim: In the present study, we determined the effect and optimum level of inclusion of a thermo-tolerant probiotic Saccharomyces cerevisiae OBV9 on nutrient intake and digestibilities in Murrah buffaloes (Bubalus bubalis. Materials and Methods: A straw based complete diet (crude protein (CP 12% with 0.1, 0.2 and 0.3% of lyophilized yeast was evaluated in a 4X4 Latin Square metabolism trial in fistulated Murrah buffalo steers (223.81 kg ± 2.91. Diet with no yeast was taken as control. Results: Dry matter intake (DMI was not significantly (P>0.05 influenced by any level (% of yeast in complete diet. The digestibility's (% of dry matter (DM and crude fibre (CF were significantly (P0.05 different from each other. Intake of DCP (g/d or g/kg w0.75, TDN (kg/d or g/kg w0.75, ME (MJ/d or MJ/kg w0.75 and N balance (g/d were not (P>0.05 affected by the level of yeast in the diet. Conclusion: It can be concluded that, 0.1% thermo-tolerant yeast had greater influence on majority of the nutrients in the straw based complete diets of Murrah buffaloes.

  11. Viability of Sea Fan Populations Impacted by Disease: Recruitment versus Incidence

    Directory of Open Access Journals (Sweden)

    Alberto M. Sabat

    2015-01-01

    Full Text Available Diseases are a threat to the viability of an increasing number of coral populations. In this study we analyze the effect of infection and recruitment rate on the viability and structure of sea fan (Gorgonia ventalina populations using a size-based matrix model parameterized with data from field studies. The model predicts that the viability of sea fan populations is strongly influenced by disease incidence and recruitment. Under high recruitment rate, the disease incidence threshold for population viability is 0.12/yr. However, populations with no or low incidence may also go locally extinct given persistent low recruitment. The model also predicts an effect of recruitment on disease prevalence. Everything else being equal, sites with low recruitment will exhibit higher disease prevalence than ones with high recruitment, particularly in medium and large colonies. Elasticity analysis reveals that changes in colony survivorship are likely to have the strongest effect on population growth rate, particularly given low recruitment. We conclude that under current levels of incidence sea fan populations in the Caribbean are not at risk. However, future epizootics are likely to produce local extinctions particularly if coinciding spatially or temporally with low recruitment.

  12. Yes-associated protein homolog, YAP-1, is involved in the thermotolerance and aging in the nematode Caenorhabditis elegans

    Energy Technology Data Exchange (ETDEWEB)

    Iwasa, Hiroaki [Department of Medical Biochemistry, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo 113-8519 (Japan); Maimaiti, Sainawaer [Department of Medical Biochemistry, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo 113-8519 (Japan); Department of Psychotherapy, The Fourth People' s Hospital of Urumqi, Urumqi 830000 (China); Kuroyanagi, Hidehito [Laboratory of Gene Expression, Graduate School of Biomedical Science, Tokyo Medical and Dental University, Tokyo 113-8519 (Japan); Kawano, Shodai; Inami, Kazutoshi; Timalsina, Shikshya; Ikeda, Mitsunobu; Nakagawa, Kentaro [Department of Medical Biochemistry, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo 113-8519 (Japan); Hata, Yutaka, E-mail: yuhammch@tmd.ac.jp [Department of Medical Biochemistry, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo 113-8519 (Japan)

    2013-04-15

    The mammalian Hippo pathway comprises mammalian Ste20-like kinases (MST1/2) and large tumor suppressor kinases (LATS1/2). LATS1/2, which are activated by MST1/2, phosphorylate a transcriptional co-activator, yes-associated protein (YAP), and induce the recruitment of YAP by 14-3-3 to cytoplasm, so that the TEAD-dependent gene transcriptions are turned off. Although the core components of the Hippo pathway are well conserved in metazoans, it has been discussed that Caenorhabditis elegans lacks YAP ortholog, we found that F13E6.4 gene encodes a protein that shows sequence similarities to YAP in the N-terminal TEAD-binding domain and in the WW domain. We designated this gene as yap-1. YAP-1 is widely expressed in various cells such as epithelial cells, muscles, hypodermal cells, gonadal sheath cells, spermatheca, and hypodermal cells. YAP-1 is distributed in cytoplasm and nuclei. wts-1 (LATS ortholog) and ftt-2 (14-3-3 ortholog) knockdowns cause nuclear accumulation of YAP-1, supporting that the subcellular localization of YAP-1 is regulated in a similar way as that of YAP. Heat shock also causes the nuclear accumulation of YAP-1 but after heat shock, YAP-1 translocates to cytoplasm. Knockdowns of DAF-21 (HSP90 ortholog) and HSF-1block the nuclear export of YAP-1 during this recovery. YAP-1 overexpression is beneficial for thermotolerance, whereas YAP-1 hyperactivity induced by wts-1 and ftt-2 knockdowns is deleterious on thermal response and yap-1 deficiency promotes health aging. In short, YAP-1 partially shares basal characters with mammalian YAP and plays a role in thermal stress response and healthy aging. - Highlights: ► We named Caenorhabditis elegans F13E6.4 gene yap-1 as a putative YAP homolog. ► The localization of YAP-1 is regulated by WTS-1 and FTT-2. ► YAP-1 is involved in healthy aging and thermosensitivity.

  13. Transcriptional activator Cat8 is involved in regulation of xylose alcoholic fermentation in the thermotolerant yeast Ogataea (Hansenula) polymorpha.

    Science.gov (United States)

    Ruchala, Justyna; Kurylenko, Olena O; Soontorngun, Nitnipa; Dmytruk, Kostyantyn V; Sibirny, Andriy A

    2017-02-28

    Efficient xylose alcoholic fermentation is one of the key to a successful lignocellulosic ethanol production. However, regulation of this process in the native xylose-fermenting yeasts is poorly understood. In this work, we paid attention to the transcriptional factor Cat8 and its possible role in xylose alcoholic fermentation in Ogataea (Hansenula) polymorpha. In Saccharomyces cerevisiae, organism, which does not metabolize xylose, gene CAT8 encodes a Zn-cluster transcriptional activator necessary for expression of genes involved in gluconeogenesis, respiration, glyoxylic cycle and ethanol utilization. Xylose is a carbon source that could be fermented to ethanol and simultaneously could be used in gluconeogenesis for hexose synthesis. This potentially suggests involvement of CAT8 in xylose metabolism. Here, the role of CAT8 homolog in the natural xylose-fermenting thermotolerant yeast O. polymorpha was characterized. The CAT8 ortholog was identified in O. polymorpha genome and deleted both in the wild-type strain and in advanced ethanol producer from xylose. Constructed cat8Δ strain isolated from wild strain showed diminished growth on glycerol, ethanol and xylose as well as diminished respiration on the last substrate. At the same time, cat8Δ mutant isolated from the best available O. polymorpha ethanol producer showed only visible defect in growth on ethanol. CAT8 deletant was characterized by activated transcription of genes XYL3, DAS1 and RPE1 and slight increase in the activity of several enzymes involved in xylose metabolism and alcoholic fermentation. Ethanol production from xylose in cat8Δ mutants in the background of wild-type strain and the best available ethanol producer from xylose increased for 50 and 30%, respectively. The maximal titer of ethanol during xylose fermentation was 12.5 g ethanol/L at 45 °C. Deletion of CAT8 did not change ethanol production from glucose. Gene CAT8 was also overexpressed under control of the strong constitutive

  14. The market viability of nuclear hydrogen technologies.

    Energy Technology Data Exchange (ETDEWEB)

    Botterud, A.; Conzelmann, G.; Petri, M. C.; Yildiz, B.

    2007-04-06

    The Department of Energy Office of Nuclear Energy is supporting system studies to gain a better understanding of nuclear power's potential role in a hydrogen economy and what hydrogen production technologies show the most promise. This assessment includes identifying commercial hydrogen applications and their requirements, comparing the characteristics of nuclear hydrogen systems to those market requirements, evaluating nuclear hydrogen configuration options within a given market, and identifying the key drivers and thresholds for market viability of nuclear hydrogen options. One of the objectives of the current analysis phase is to determine how nuclear hydrogen technologies could evolve under a number of different futures. The outputs of our work will eventually be used in a larger hydrogen infrastructure and market analysis conducted for DOE-EE using a system-level market simulation tool now underway. This report expands on our previous work by moving beyond simple levelized cost calculations and looking at profitability, risk, and uncertainty from an investor's perspective. We analyze a number of technologies and quantify the value of certain technology and operating characteristics. Our model to assess the profitability of the above technologies is based on Real Options Theory and calculates the discounted profits from investing in each of the production facilities. We use Monte-Carlo simulations to represent the uncertainty in hydrogen and electricity prices. The model computes both the expected value and the distribution of discounted profits from a production plant. We also quantify the value of the option to switch between hydrogen and electricity production in order to maximize investor profits. Uncertainty in electricity and hydrogen prices can be represented with two different stochastic processes: Geometric Brownian Motion (GBM) and Mean Reversion (MR). Our analysis finds that the flexibility to switch between hydrogen and electricity leads

  15. Puget Sound steelhead life cycle model analyses - Population Viability Analysis

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This research was initiated by the Puget Sound Steelhead Technical Recovery Team to develop viability criteria for threatened Puget Sound steelhead and to support...

  16. Cadmium affects the mitochondrial viability and the acid soluble ...

    African Journals Online (AJOL)

    Cadmium affects the mitochondrial viability and the acid soluble thiols concentration in liver, kidney, heart and gills of Ancistrus brevifilis (Eigenmann, 1920). P Velasquez-Vottelerd, Y Anton, R Salazar-Lugo ...

  17. Maintaining yeast viability in continuous primary beer fermentation

    National Research Council Canada - National Science Library

    Pires, Eduardo J; Teixeira, José A; Brányik, Tomás; Côrte‐Real, Manuela; Vicente, António A

    2014-01-01

    .... This work was aimed at solving one of the most relevant obstacles to implementing ICT on a large scale in beer fermentations, namely the control of biomass and the maintenance of cell viability in a gas‐lift bioreactor...

  18. Desiccation-induced changes in viability, lipid peroxidation and ...

    African Journals Online (AJOL)

    user

    2012-05-31

    Hendry et al., 1992) and A. saccharinum (Pukacka and Ratajczak, 2006) and intermediate seeds like Azadirachta indica (Varghese and. Naithani, 2002), Coffea Arabica (Dussert et al., 2006), indicating that loss of seed viability ...

  19. Femtosecond optical transfection of cells:viability and efficiency

    National Research Council Canada - National Science Library

    D. Stevenson; B. Agate; X. Tsampoula; P. Fischer; C. T. A. Brown; W. Sibbett; A. Riches; F. Gunn-Moore; K. Dholakia

    2006-01-01

    .... However, there remains no study into the true efficiency of this procedure. Here, we present a detailed analysis of transfection efficiency and cell viability for femtosecond optical transfection using a titanium sapphire laser at 800 nm...

  20. Equine ovarian tissue viability after cryopreservation and in vitro culture

    Science.gov (United States)

    The efficiency of several cryoprotective agents were compared using both slow-freezing and vitrification methods. Results indicate that the viability of ovarian tissue cells increases when DMSO (slow-freezing) and ethylene glycol (vitrification) are used....

  1. Approximate viability for nonlinear evolution inclusions with application to controllability

    Directory of Open Access Journals (Sweden)

    Omar Benniche

    2016-12-01

    Full Text Available We investigate approximate viability for a graph with respect to fully nonlinear quasi-autonomous evolution inclusions. As application, an approximate null controllability result is given.

  2. Characterization of Thermotolerant Chitinases Encoded by a Brevibacillus laterosporus Strain Isolated from a Suburban Wetland

    Directory of Open Access Journals (Sweden)

    Pulin Liu

    2015-12-01

    Full Text Available To isolate and characterize chitinases that can be applied with practical advantages, 57 isolates of chitin-degrading bacteria were isolated from the soil of a suburban wetland. 16S rRNA gene analysis revealed that the majority of these strains belonged to two genera, Paenibacillus and Brevibacillus. Taking thermostability into account, the chitinases (ChiA and ChiC of a B. laterosporus strain were studied further. Ni-NTA affinity-purified ChiA and ChiC were optimally active at pH 7.0 and 6.0, respectively, and showed high temperature stability up to 55 °C. Kinetic analysis revealed that ChiC has a lower affinity and stronger catalytic activity toward colloidal chitin than ChiA. With their stability in a broad temperature range, ChiA and ChiC can be utilized for the industrial bioconversion of chitin wastes into biologically active products.

  3. Novel Thermotolerant Siderophilic Filamentous Cyanobacterium that Produces Intracellular Iron-Rich Phases

    Science.gov (United States)

    Broun, Igor I.; Bryant, Donald A.; Casamatta, Dale; Thomas-Keprta, Kathie L.; Sarkisova, Svetlana A.; Shen, Gaozhang; Graham, Joel E.; Boyd, Eric S.; Peters, John W.; Garrison, Daniel H.; hide

    2010-01-01

    Cyanobacteria are the main producers of organic compounds in iron-depositing hot springs despite photosynthetically generated-oxygen and the abundance of reduced iron (Fe2+) that likely leads to enormous oxidative stress within cyanobacterial cells. Therefore, the study of cyanobacterial diversity, phylogeny, and biogeochemical activity in iron-depositing hot springs will not only provide insights into the contribution of CB to iron redox cycling in these environments, but it could also provide insights into CB evolution. This study characterizes the phylogeny, morphology, and physiology of isolate JSC-1, a novel filamentous CB isolated from an iron-depositing hot spring. While isolate JSC-1 is morphologically similar to the CB genus Leptolyngbya, 16S rDNA sequence data indicated that it shares 95 percent sequence similarity to the type strain L. boryanum. Strain JSC-1 fixes N2 and exhibited an unusually high ratio between photosystem (PS) I and PS II and was capable of complementary chromatic adaptation. Further, it synthesized only chlorophyll a and a unique set of carotenoids. Strain JSC-1 not only required high levels of Fe for growth (greater than or equal to 40 microM), but it also accumulated large amounts of extracellular ferrihydrite and generated intracellular ferric phosphates. Strain JSC-1 was found to secrete 2-oxoglutaric acid and possesses one ortholog and one paralog of bacterioferritin. Surprisingly, the latter has 70.13 % identity with a bacterioferritin in marine-proteobacterium HTCC 2080 and has joint node with bacterioferritins found in enterobacteria. Collectively, these observations provide insights into the physiological strategies that might have allowed CB to develop and proliferate in Fe-rich environments. Based on its genotypic and phenotypic characterization of strain, JSC-1 represents a new operational taxonomical unit (OTU) JSC-1.

  4. Myocardial viability estimation during the recovery phase of stress echocardiography after acute beta-blocker administration.

    Science.gov (United States)

    Karagiannis, Stefanos E; Feringa, Harm H H; Bax, Jeroen J; Elhendy, Abdu; Dunkelgrun, Martin; Vidakovic, Radosav; Hoeks, S E; van Domburg, Ron; Valhema, Roelf; Cokkinos, Dennis V; Poldermans, Don

    2007-04-01

    Myocardial viability assessment in severely dysfunctional segments by dobutamine stress echocardiography (DSE) is less sensitive than nuclear scanning. To assess the additional value of using the recovery phase of DSE after acute beta-blocker administration for identifying viable myocardium. The study included 49 consecutive patients with ejection fraction (LVEF)DISA-SPECT) evaluation for viability of severely dysfunctional segments. Patients with >or=4 viable segments were considered viable. Coronary revascularization followed within 3 months in all patients. Radionuclide evaluation of LVEF was performed before and 12 months after revascularization. Viability with DISA-SPECT was detected in 463 (59%) segments, while 154 (19.7%) segments presented as scar. The number of viable segments increased from 415 (53%) at DSE to 463 (59%) at DSE and recovery, and the number of viable patients increased from 43 to 49 respectively. LVEF improved by >or=5% in 27 patients. Multivariate regression analysis showed that, DSE with recovery phase was the only independent predictor of >or=5% LVEF improvement after revascularization (OR 14.6, CI 1.4-133.7). In this study, we demonstrate that the recovery phase of DSE has an increased sensitivity for viability estimation compared to low-high dose DSE.

  5. Ovarian fluid mediates the temporal decline in sperm viability in a fish with sperm storage.

    Directory of Open Access Journals (Sweden)

    Clelia Gasparini

    Full Text Available A loss of sperm viability and functionality during sperm transfer and storage within the female reproductive tract can have important fitness implications by disrupting fertilization and impairing offspring development and survival. Consequently, mechanisms that mitigate the temporal decline in sperm function are likely to be important targets of selection. In many species, ovarian fluid is known to regulate and maintain sperm quality. In this paper, we use the guppy Poecilia reticulata, a highly polyandrous freshwater fish exhibiting internal fertilization and sperm storage, to determine whether ovarian fluid (OF influences the decline in sperm viability (the proportion of live sperm in the ejaculate over time and whether any observed effects depend on male sexual ornamentation. To address these questions we used a paired experimental design in which ejaculates from individual males were tested in vitro both in presence and absence of OF. Our results revealed that the temporal decline in sperm viability was significantly reduced in the presence of OF compared to a saline control. This finding raises the intriguing possibility that OF may play a role in mediating the decline in sperm quality due to the deleterious effects of sperm ageing, although other possible explanations for this observation are discussed. Interestingly, we also show that the age-related decline in sperm viability was contingent on male sexual ornamentation; males with relatively high levels of iridescence (indicating higher sexual attractiveness exhibited a more pronounced decline in sperm viability over time than their less ornamented counterparts. This latter finding offers possible insights into the functional basis for the previously observed trade-off between these key components of pre- and postcopulatory sexual selection.

  6. Thermotolerant and mesophylic fungi from sugarcane bagasse and their prospection for biomass-degrading enzyme production

    Directory of Open Access Journals (Sweden)

    Bruna Silveira Lamanes dos Santos

    2015-09-01

    Full Text Available Nineteen fungi and seven yeast strains were isolated from sugarcane bagasse piles from an alcohol plant located at Brazilian Cerrado and identified up to species level on the basis of the gene sequencing of 5.8S-ITS and 26S ribosomal DNA regions. Four species were identified: Kluyveromyces marxianus, Aspergillus niger, Aspergillus sydowii and Aspergillus fumigatus, and the isolates were screened for the production of key enzymes in the saccharification of lignocellulosic material. Among them, three strains were selected as good producers of hemicellulolitic enzymes: A. niger (SBCM3, A. sydowii (SBCM7 and A. fumigatus (SBC4. The best β-xylosidase producer was A. niger SBCM3 strain. This crude enzyme presented optimal activity at pH 3.5 and 55 °C (141 U/g. For β-glucosidase and xylanase the best producer was A. fumigatus SBC4 strain, whose enzymes presented maximum activity at 60 °C and pH 3.5 (54 U/g and 4.0 (573 U/g, respectively. All these crude enzymes presented stability around pH 3.0–8.0 and up to 60 °C, which can be very useful in industrial processes that work at high temperatures and low pHs. These enzymes also exhibited moderate tolerance to ethanol and the sugars glucose and xylose. These similar characteristics among these fungal crude enzymes suggest that they can be used synergistically in cocktails in future studies of biomass conversion with potential application in several biotechnological sectors.

  7. The viability of native microbial communities in martian environment (model)

    Science.gov (United States)

    Vorobyova, Elena; Cheptcov, Vladimir; Pavlov, Anatolyi; Vdovina, Mariya; Lomasov, Vladimir

    For today the important direction in astrobiology is the experimental simulation of extraterrestrial habitats with the assessment of survivability of microorganisms in such conditions. A new task is to investigate the resistance of native microbial ecosystems which are well adapted to the environment and develop unique protection mechanisms that enable to ensure biosphere formation. The purpose of this research was to study the viability of microorganisms as well as viability of native microbial communities of arid soils and permafrost under stress conditions simulating space environment and martian regolith environment, estimation of duration of Earth like life in the Martian soil. The experimental data obtained give the proof of the preservation of high population density, biodiversity, and reproduction activity under favorable conditions in the Earth analogues of Martian soil - arid soils (Deserts of Israel and Morocco) and permafrost (East Siberia, Antarctica), after the treatment of samples by ionizing radiation dose up to 100 kGy at the pressure of 1 torr, temperature (- 50oC) and in the presence of perchlorate (5%). It was shown that in simulated conditions close to the parameters of the Martian regolith, the diversity of natural bacterial communities was not decreased, and in some cases the activation of some bacterial populations occurred in situ. Our results allow suggesting that microbial communities like those that inhabit arid and permafrost ecosystems on the Earth, can survive at least 500 thousand years under conditions of near surface layer of the Martian regolith. Extrapolation of the data according to the intensity of ionizing radiation to the open space conditions allows evaluating the potential lifespan of cells inside meteorites as 20-50 thousand years at least. In this work new experimental data have been obtained confirming the occurrence of liquid water and the formation of wet soil layer due to sublimation of ice when the temperature of the

  8. Viability of microencapsulated Lactobacillus casei in synbiotic mayonnaise

    Directory of Open Access Journals (Sweden)

    Lieu, M.D.

    2017-07-01

    Full Text Available In this study, whey protein, maltodextrin and GOS (Galacto-oligosaccharides used as microencapsulating agents to protect Lactobacillus casei during spray-drying and mayonnaise storage. The morphology of microcapsules, pH charges, the survival rate during mayonnaise storage as well as survival in simulated gastric fluid (SGF and intestinal fluid (SIF was tested in this study. The results indicated that whey protein showed a protective effect better than maltodextrin during spray-drying. The particles showed spherical shape and typical concavity of all samples and encapsulating agents were not affected by the size and surface structure of particles. The pH charges were not significantly different in all mayonnaise samples in this test. The viability of free cell L. casei after 6 weeks storage was significant decrease about 4 log CFU/g compared to 1.55 to 3.27 log CFU/g in the mayonnaise samples containing microcapsules in which maltodextrin showed the lowest of L. casei survival rate. In SGF and SIF conditions, maltodextrin act as prebiotic sufficiently which do not need adding GOS. The combination of whey protein and maltodextrin in which maltodextrin plays a role as supporting agents for the spray-drying process as well as prebiotic potential, while whey protein with high buffer property which enhancing the survival rate of L. casie in low pH.

  9. Wastewater treatment to enhance the economic viability of microalgae culture.

    Science.gov (United States)

    Pires, J C M; Alvim-Ferraz, M C M; Martins, F G; Simões, M

    2013-08-01

    Microalgae culture is still not economically viable and it presents some negative environmental impacts, concerning water, nutrient and energy requirements. In this context, this study aims to review the recent advances on microalgal cultures in wastewaters to enhance their economic viability. We focused on three different culture concepts: (1) suspended cell systems, (2) cell immobilization, and (3) microalgae consortia. Cultures with suspended cells are the most studied. The nutrient removal efficiencies are usually high for wastewaters of different sources. However, biomass harvesting is difficult and a costly process due to the small cell size and lower culture density. On the other hand, the cell immobilization systems showed to be the solution for this problem, having as main limitation the nutrient diffusion from bulk to cells, which results in a reduced nutrient removal efficiency. The consortium between microalgae and bacteria enhances the growth of both microorganisms. This culture concept showed to be a promising technology to improve wastewater treatment, regarding not only nutrient removal but also biomass harvesting by bioflocculation. The aggregation mechanism must be studied in depth to find the process parameters that would lead to an effective and cheap harvesting process.

  10. Comparison of reintroduction and enhancement effects on metapopulation viability

    Science.gov (United States)

    Halsey, Samniqueka J; Bell, Timothy J.; McEachern, Kathryn; Pavlovic, Noel B.

    2015-01-01

    Metapopulation viability depends upon a balance of extinction and colonization of local habitats by a species. Mechanisms that can affect this balance include physical characteristics related to natural processes (e.g. succession) as well as anthropogenic actions. Plant restorations can help to produce favorable metapopulation dynamics and consequently increase viability; however, to date no studies confirm this is true. Population viability analysis (PVA) allows for the use of empirical data to generate theoretical future projections in the form of median time to extinction and probability of extinction. In turn, PVAs can inform and aid the development of conservation, recovery, and management plans. Pitcher's thistle (Cirsium pitcheri) is a dune endemic that exhibited metapopulation dynamics. We projected viability of three natural and two restored populations with demographic data spanning 15–23 years to determine the degree the addition of reintroduced population affects metapopulation viability. The models were validated by comparing observed and projected abundances and adjusting parameters associated with demographic and environmental stochasticity to improve model performance. Our chosen model correctly predicted yearly population abundance for 60% of the population-years. Using that model, 50-year projections showed that the addition of reintroductions increases metapopulation viability. The reintroduction that simulated population performance in early-successional habitats had the maximum benefit. In situ enhancements of existing populations proved to be equally effective. This study shows that restorations can facilitate and improve metapopulation viability of species dependent on metapopulation dynamics for survival with long-term persistence of C. pitcheri in Indiana likely to depend on continued active management.

  11. Diammonium phosphate stimulates transcription of L-lactate dehydrogenase leading to increased L-lactate production in the thermotolerant Bacillus coagulans strain.

    Science.gov (United States)

    Sun, Lifan; Li, Yanfeng; Wang, Limin; Wang, Yanping; Yu, Bo

    2016-08-01

    Exploration of cost-effective fermentation substrates for efficient lactate production is an important economic objective. Although some organic nitrogen sources are also cheaper, inorganic nitrogen salts for lactate fermentation have additional advantages in facilitating downstream procedures and significantly improving the commercial competitiveness of lactate production. In this study, we first established an application of diammonium phosphate to replace yeast extract with a reduced 90 % nitrogen cost for a thermotolerant Bacillus coagulans strain. In vivo enzymatic and transcriptional analyses demonstrated that diammonium phosphate stimulates the gene expression of L-lactate dehydrogenase, thus providing higher specific enzyme activity in vivo and increasing L-lactic acid production. This new information provides a foundation for establishing a cost-effective process for polymer-grade L-lactic acid production in an industrial setting.

  12. Towards an international standard for PCR-based detection of foodborne thermotolerant campylobacters: interaction of enrichment media and pre-PCR treatment on carcass rinse samples

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Lübeck, Peter Stephensen; Hansen, F.

    2004-01-01

    As part of a large EU project for standardisation of polymerase chain reaction (PCR), a systematic evaluation of the interaction of enrichment media, type of DNA polymerase and pre-PCR sample treatment for a PCR detecting thermotolerant campylobacters was carried out. The growth-supporting capacity...... and PCR compatibility of enrichment in Preston, Mueller-Hinton and Bolton broth (blood-containing and blood-free) were evaluated. The effect of resin-based DNA extraction and DNA extraction by boiling on the final PCR assay was investigated. The time-course studies indicated that a 20-h sample enrichment...... in blood-containing Bolton broth, followed by a simple resin-based extraction of DNA and a PCR amplification using Tth polymerase, resulted in strong and clear PCR amplicons for target (287 bp) and internal amplification control (IAC, 124 bp). The enrichment PCR-based method, tested on 68 presumably...

  13. Adaptation to thermotolerance in Rhizopus coincides with virulence as revealed by avian and invertebrate infection models, phylogeny, physiological and metabolic flexibility.

    Science.gov (United States)

    Kaerger, Kerstin; Schwartze, Volker U; Dolatabadi, Somayeh; Nyilasi, Ildikó; Kovács, Stella A; Binder, Ulrike; Papp, Tamás; Hoog, Sybren de; Jacobsen, Ilse D; Voigt, Kerstin

    2015-01-01

    Mucormycoses are fungal infections caused by the ancient Mucorales. They are rare, but increasingly reported. Predisposing conditions supporting and favoring mucormycoses in humans and animals include diabetic ketoacidosis, immunosuppression and haematological malignancies. However, comprehensive surveys to elucidate fungal virulence in ancient fungi are limited and so far focused on Lichtheimia and Mucor. The presented study focused on one of the most important causative agent of mucormycoses, the genus Rhizopus (Rhizopodaceae). All known clinically-relevant species are thermotolerant and are monophyletic. They are more virulent compared to non-clinically, mesophilic species. Although adaptation to elevated temperatures correlated with the virulence of the species, mesophilic strains showed also lower virulence in Galleria mellonella incubated at permissive temperatures indicating the existence of additional factors involved in the pathogenesis of clinical Rhizopus species. However, neither specific adaptation to nutritional requirements nor stress resistance correlated with virulence, supporting the idea that Mucorales are predominantly saprotrophs without a specific adaptation to warm blooded hosts.

  14. Bioconversion of Saccharum spontaneum (wild sugarcane) hemicellulosic hydrolysate into ethanol by mono and co-cultures of Pichia stipitis NCIM3498 and thermotolerant Saccharomyces cerevisiae-VS₃.

    Science.gov (United States)

    Chandel, Anuj K; Singh, Om V; Narasu, M Lakshmi; Rao, L Venkateswar

    2011-10-01

    The lignocellulosic biomass is a low-cost renewable resource for eco-benign liquid fuel 'ethanol'. To resolve the hydrolysis of mixed sugars in lignocellulosic substrate Saccharum spontaneum, the microbial co-cultures of Pichia stipitis NCIM 3498 and thermotolerant Saccharomyces cerevisiae-VS(3) were analyzed for efficient bioconversion of mixed sugars into ethanol. Among the hydrolysis conditions, the acid hydrolysis released maximum sugars along with furans, phenolics and acetic acid. The acidic hydrolysate was detoxified and fermented by monocultures of P. stipitis NCIM3498 (P.S.) and thermotolerant S. cerevisiae VS(3) (S.C.), and co-culture of P.S. (7.5 mL) and S.C. (2.5 mL). Before the fermentation of hemicellulose acid hydrolysate, both the monocultures (P.S. and S.C.), and varying ratios of P.S. and S.C. microorganisms in co-cultures #1, #2 and #3 were grown on simulated synthetic medium. The ethanol yield from monocultures of P.S. (0.44 ± 0.021 g/g), S.C. (0.22 ± 0.01 g/g) and co-culture #3 (0.49 ± 0.02 g/g) revealed unique characteristics of each mono and co-culture technology. The fermentation of hemicellulose acid hydrolysate with monocultures of P.S., S.C. and co-culture #3 produced 12.08 ± 0.72 g/L, 1.40 ± 0.07 g/L, and 15.0 ± -0.92 g/L ethanol, respectively. Copyright © 2011. Published by Elsevier B.V.

  15. Importance of Donor Chondrocyte Viability for Osteochondral Allografts.

    Science.gov (United States)

    Cook, James L; Stannard, James P; Stoker, Aaron M; Bozynski, Chantelle C; Kuroki, Keiichi; Cook, Cristi R; Pfeiffer, Ferris M

    2016-05-01

    Osteochondral allograft (OCA) transplantation provides a biological treatment option for functional restoration of large articular cartilage defects in multiple joints. While successful outcomes after OCA transplantation have been linked to viable donor chondrocytes, the importance of donor cell viability has not been comprehensively validated. To use a canine model to determine the importance of donor chondrocyte viability at the time of implantation with respect to functional success of femoral condylar OCAs based on radiographic, gross, cell viability, histologic, biochemical, and biomechanical outcome measures. Controlled laboratory study. After approval was obtained from the institutional animal care and use committee, adult female dogs (N = 16) were implanted with 8-mm cylindrical OCAs from male dogs in the lateral and medial femoral condyles of 1 knee. OCAs were preserved for 28 or 60 days after procurement, and chondrocyte viability was quantified before implantation. Two different storage media, temperatures, and time points were used to obtain a spectrum of percentage chondrocyte viability at the time of implantation. A successful outcome was defined as an OCA that was associated with graft integration, maintenance of hyaline cartilage, lack of associated cartilage disorder, and lack of fibrillation, fissuring, or fibrous tissue infiltration of the allograft based on subjective radiographic, gross, and histologic assessments at 6 months after implantation. Chondrocyte viability ranged from 23% to 99% at the time of implantation. All successful grafts had >70% chondrocyte viability at the time of implantation, and no graft with chondrocyte viability <70% was associated with a successful outcome. Live-dead stained sections and histologic findings with respect to cell morphological features suggested that successful grafts were consistently composed of viable chondrocytes in lacunae, while grafts that were not successful were composed of nonviable

  16. On the Viability of Conspiratorial Beliefs

    OpenAIRE

    David Robert Grimes

    2016-01-01

    Conspiratorial ideation is the tendency of individuals to believe that events and power relations are secretly manipulated by certain clandestine groups and organisations. Many of these ostensibly explanatory conjectures are non-falsifiable, lacking in evidence or demonstrably false, yet public acceptance remains high. Efforts to convince the general public of the validity of medical and scientific findings can be hampered by such narratives, which can create the impression of doubt or ...

  17. A Classification Method for Seed Viability Assessment with Infrared Thermography

    Directory of Open Access Journals (Sweden)

    Sen Men

    2017-04-01

    Full Text Available This paper presents a viability assessment method for Pisum sativum L. seeds based on the infrared thermography technique. In this work, different artificial treatments were conducted to prepare seeds samples with different viability. Thermal images and visible images were recorded every five minutes during the standard five day germination test. After the test, the root length of each sample was measured, which can be used as the viability index of that seed. Each individual seed area in the visible images was segmented with an edge detection method, and the average temperature of the corresponding area in the infrared images was calculated as the representative temperature for this seed at that time. The temperature curve of each seed during germination was plotted. Thirteen characteristic parameters extracted from the temperature curve were analyzed to show the difference of the temperature fluctuations between the seeds samples with different viability. With above parameters, support vector machine (SVM was used to classify the seed samples into three categories: viable, aged and dead according to the root length, the classification accuracy rate was 95%. On this basis, with the temperature data of only the first three hours during the germination, another SVM model was proposed to classify the seed samples, and the accuracy rate was about 91.67%. From these experimental results, it can be seen that infrared thermography can be applied for the prediction of seed viability, based on the SVM algorithm.

  18. Viability of Cryptosporidium parvum during ensilage of perennial ryegrass.

    Science.gov (United States)

    Merry, R J; Mawdsley, J L; Brooks, A E; Davies, D R

    1997-01-01

    The survival of Cryptosporidium parvum during ensilage of perennial ryegrass was examined in laboratory silos with herbage prepared in one of three different ways; either untreated, inoculated with a strain of Lactobacillus plantarum or by direct acidification with formic acid. The pH values of all silages initially fell below 4.5, but only formic acid-treated silage remained stable at less than pH 4 after 106 d, with the pH of the untreated and inoculant-treated silages rising to above 6. The formic acid-treated silage had a high lactic acid concentration (109 g kg-1 dry matter (DM)) and low concentrations of propionic and butyric acids after 106 d. However, the untreated and inoculant-treated silages showed an inverse relationship, with low lactic acid concentrations and high concentrations of acetic, propionic and butyric acids. These silages also contained ammonia-N concentrations in excess of 9 g kg-1 DM. In terms of the viability of Cryptosporidium parvum oocysts very few differences were seen after 14 d of ensilage with ca 50% remaining viable, irrespective of treatment and total numbers had declined from the initial level of 5.9 x 10(4) to 1 x 10(4) g(-1) fresh matter. Total oocyst numbers remained approximately the same until the end of the ensiling period, with the percentage of viable oocysts declining to 46, 41 and 32% respectively for formic acid, inoculant and untreated silages. The results are discussed in terms of changes occurring during the silage fermentation, in particular the products which may influence the survival of Cryptosporidium and implications for agricultural practice and the health of silage fed livestock.

  19. Regional population viability of grassland songbirds: Effects of agricultural management

    Science.gov (United States)

    Perlut, N.G.; Strong, A.M.; Donovan, T.M.; Buckley, N.J.

    2008-01-01

    Although population declines of grassland songbirds in North America and Europe are well-documented, the effect of local processes on regional population persistence is unclear. To assess population viability of grassland songbirds at a regional scale (???150,000 ha), we quantified Savannah Sparrow Passerculus sandwichensis and Bobolink Dolichonyx oryzivorus annual productivity, adult apparent survival, habitat selection, and density in the four most (regionally) common grassland treatments. We applied these data to a female-based, stochastic, pre-breeding population model to examine whether current grassland management practices can sustain viable populations of breeding songbirds. Additionally, we evaluated six conservation strategies to determine which would most effectively increase population trends. Given baseline conditions, over 10 years, simulations showed a slightly declining or stable Savannah Sparrow population (mean bootstrap ?? = 0.99; 95% CI = 1.00-0.989) and severely declining Bobolink population (mean bootstrap ?? = 0.75; 95% CI = 0.753-0.747). Savannah Sparrow populations were sensitive to increases in all demographic parameters, particularly adult survival. However for Bobolinks, increasing adult apparent survival, juvenile apparent survival, or preference by changing habitat selection cues for late-hayed fields (highest quality) only slightly decreased the rate of decline. For both species, increasing the amount of high-quality habitat (late- and middle-hayed) marginally slowed population declines; increasing the amount of low-quality habitat (early-hayed and grazed) marginally increased population declines. Both species were most sensitive to low productivity and survival on early-hayed fields, despite the fact that this habitat comprised only 18% of the landscape. Management plans for all agricultural regions should increase quality on both low- and high-quality fields by balancing habitat needs, nesting phenology, and species' response to

  20. Tissue viability 2010 -2015:from good to great.

    Science.gov (United States)

    Milne, Jeanette; Ousey, Karen

    2010-09-01

    This paper explores the challenges of the changing face of the NHS with specific relation to the challenges for community-based tissue viability services following the publication of government documents that identify the need to provide a quality service for all patients in health-care settings. Patients receiving care in the community is paramount to the success of the NHS going forward; service redesign, improvements in quality, outcome tracking, seamless discharge and patient satisfaction/responsibilities has been heralded as the core prerequisites of successful services. Tissue viability is a relatively young specialism, with most services being nurse led and established less than 15 years. It is argued that in order to continue to be successful as a specialism, tissue viability has to challenge traditional patient and nursing beliefs and values.

  1. Viability of Lucilia sericata maggots after exposure to wound antiseptics.

    Science.gov (United States)

    Daeschlein, Georg; Napp, Matthias; Assadian, Ojan; von Podewils, Sebastian; Reese, Kevin; Hinz, Peter; Matiasek, Johannes; Spitzmueller, Romy; Humphreys, Paul; Jünger, Michael; Kramer, Axel

    2017-06-01

    After debridement and before dressing a wound with maggots of calliphorid flies, one frequently performed step is the application of antiseptics to the prepared wound bed. However, the concomitant application of antiseptic agents during maggot therapy is regarded controversial as antiseptics may interfere with maggots' viability. In this experimental in vitro study, the viability of fly maggots was investigated after exposure to various antiseptics frequently used in wound care. Here, we show that Lucilia sericata fly maggots can survive up to an hour's exposure to wound antiseptics such as octenidine, povidone-iodine or polihexanide. Concomitant short-term application of wound antiseptics together with maggots on wound beds is tolerated by larvae and does not impair their viability. © 2016 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  2. Economic Viability of Brewery Spent Grain as a Biofuel

    Energy Technology Data Exchange (ETDEWEB)

    Morrow, Charles [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2016-01-01

    This report summarizes an investigation into the technical feasibility and economic viability of use grain wastes from the beer brewing process as fuel to generate the heat needed in subsequent brewing process. The study finds that while use of spent grain as a biofuel is technically feasible, the economics are not attractive. Economic viability is limited by the underuse of capital equipment. The investment in heating equipment requires a higher utilization that the client brewer currently anticipates. It may be possible in the future that changing factors may swing the decision to a more positive one.

  3. Economic viability of new launched school lunch programmes

    DEFF Research Database (Denmark)

    Jensen, Jørgen Dejgård; Smed, Sinne; Mørkbak, Morten Raun

    2013-01-01

    The objective of this paper is to investigate determinants for the viability of school lunch programmes with a zero-price start-up period. The study is based on a Danish pilot experiment, in which 38 schools were subsidized to provide free school lunch for all pupils during a two-month start...... activities related to the schools’ support and the users’ feeling of ownership, as well as internal professionalism and leadership in the implementation of the school lunch programme are important for the viability of the programme. Strong performance on the latter factors might to some extent compensate...... for the gap between cost and users’ willingness to pay for school lunches....

  4. Challenge testing of gametes to enhance their viability

    DEFF Research Database (Denmark)

    Callesen, Henrik

    2010-01-01

    of survival mechanism that enables them to come through the process. The details of the mechanism remain unknown but, if identified, it could have immense potential as a new way to improve the viability of embryos produced by ART. However, few publications describe systematic ways to challenge test gametes...... and then to use the results as a basis for improving gamete viability. Furthermore, new methods to monitor the reactions of gametes to such challenge tests are needed. In the present review, these two issues are discussed, as are some of the conditions necessary before a challenge test protocol can be part...

  5. Effect of culture age, protectants, and initial cell concentration on viability of freeze-dried cells of Metschnikowia pulcherrima.

    Science.gov (United States)

    Spadaro, Davide; Ciavorella, Annalisa Alessandra; Lopez-Reyes, Jorge Giovanny; Garibaldi, Angelo; Gullino, Maria Lodovica

    2010-10-01

    The effect of freeze-drying using different lyoprotectants at different concentrations on the viability and biocontrol efficacy of Metschnikowia pulcherrima was evaluated. The effects of initial yeast cell concentration and culture age on viability were also considered. Yeast cells grown for 36 h were more resistant to freeze-drying than were 48 h cells. An initial concentration of 10⁸ cells·mL⁻¹ favoured the highest survival after freeze-drying. When maltose (25%, m/v) was used as protectant, a high cell viability was obtained (64.2%). Cells maintained a high viability after 6 months of storage at 4 °C. The biocontrol efficacy of freeze-dried cells was similar to the activity of fresh cells on 'Gala' apples and was slightly lower on 'Golden Delicious' apples. After optimizing freeze-drying conditions, the viability of M. pulcherrima cells was similar to that obtained in other studies. The results constitute a first step towards the commercial development of M. pulcherrima as a biocontrol agent.

  6. Viable cell yield from active dry yeast products and effects of storage temperature and diluent on yeast cell viability.

    Science.gov (United States)

    Sullivan, M L; Bradford, B J

    2011-01-01

    Active dry yeast (ADY) products are commonly fed in the dairy industry, but research regarding quality control for such products is limited. The objectives of this study were to determine yeast viability in field samples relative to manufacturers' guarantees (experiment 1), measure the effects of high-temperature storage on yeast viability (experiment 1), and determine the effect of vitamin-trace mineral (VTM) premix on yeast viability (experiment 2). Commercially available ADY products were acquired in triplicate through normal distribution channels and stored at 4°C upon receipt. Initial samples were evaluated for colony-forming units and compared with product label guarantees. Only 1 of the 6 products sampled in experiment 1 met product guarantees for all 3 samples. To determine effects of storage temperature and duration on viability, ADY samples were stored in an incubator at 40°C with ambient humidity for 1, 2, and 3 mo. High-temperature storage significantly decreased viability over the 3-mo period; approximately 90% of viable cells were lost each month. Three of the 5 products sampled in experiment 2 met product guarantees. Fresh samples of 4 of these 5 ADY products were mixed in duplicate with ground corn (GC) or a VTM premix to achieve a target concentration of 2.2×10(8) cfu/g. For each product, GC and VTM samples were stored at ambient temperature (22°C) and at an elevated temperature (40°C) for 2 wk. No differences in viable yeast count were observed between GC and VTM samples immediately after mixing or after storage at ambient temperature. Yeast viability in GC and VTM samples decreased during storage at an elevated temperature. There also was a significant interaction of diluent and storage temperature; VTM samples had higher cell viability than GC samples when subjected to high-temperature storage. Results suggest that (1) ADY products failed to consistently meet product guarantees; (2) viability of ADY products was greatly diminished during

  7. On the Viability of Conspiratorial Beliefs

    Science.gov (United States)

    Grimes, David Robert

    2016-01-01

    Conspiratorial ideation is the tendency of individuals to believe that events and power relations are secretly manipulated by certain clandestine groups and organisations. Many of these ostensibly explanatory conjectures are non-falsifiable, lacking in evidence or demonstrably false, yet public acceptance remains high. Efforts to convince the general public of the validity of medical and scientific findings can be hampered by such narratives, which can create the impression of doubt or disagreement in areas where the science is well established. Conversely, historical examples of exposed conspiracies do exist and it may be difficult for people to differentiate between reasonable and dubious assertions. In this work, we establish a simple mathematical model for conspiracies involving multiple actors with time, which yields failure probability for any given conspiracy. Parameters for the model are estimated from literature examples of known scandals, and the factors influencing conspiracy success and failure are explored. The model is also used to estimate the likelihood of claims from some commonly-held conspiratorial beliefs; these are namely that the moon-landings were faked, climate-change is a hoax, vaccination is dangerous and that a cure for cancer is being suppressed by vested interests. Simulations of these claims predict that intrinsic failure would be imminent even with the most generous estimates for the secret-keeping ability of active participants—the results of this model suggest that large conspiracies (≥1000 agents) quickly become untenable and prone to failure. The theory presented here might be useful in counteracting the potentially deleterious consequences of bogus and anti-science narratives, and examining the hypothetical conditions under which sustainable conspiracy might be possible. PMID:26812482

  8. On the Viability of Conspiratorial Beliefs.

    Directory of Open Access Journals (Sweden)

    David Robert Grimes

    Full Text Available Conspiratorial ideation is the tendency of individuals to believe that events and power relations are secretly manipulated by certain clandestine groups and organisations. Many of these ostensibly explanatory conjectures are non-falsifiable, lacking in evidence or demonstrably false, yet public acceptance remains high. Efforts to convince the general public of the validity of medical and scientific findings can be hampered by such narratives, which can create the impression of doubt or disagreement in areas where the science is well established. Conversely, historical examples of exposed conspiracies do exist and it may be difficult for people to differentiate between reasonable and dubious assertions. In this work, we establish a simple mathematical model for conspiracies involving multiple actors with time, which yields failure probability for any given conspiracy. Parameters for the model are estimated from literature examples of known scandals, and the factors influencing conspiracy success and failure are explored. The model is also used to estimate the likelihood of claims from some commonly-held conspiratorial beliefs; these are namely that the moon-landings were faked, climate-change is a hoax, vaccination is dangerous and that a cure for cancer is being suppressed by vested interests. Simulations of these claims predict that intrinsic failure would be imminent even with the most generous estimates for the secret-keeping ability of active participants-the results of this model suggest that large conspiracies (≥1000 agents quickly become untenable and prone to failure. The theory presented here might be useful in counteracting the potentially deleterious consequences of bogus and anti-science narratives, and examining the hypothetical conditions under which sustainable conspiracy might be possible.

  9. On the Viability of Conspiratorial Beliefs.

    Science.gov (United States)

    Grimes, David Robert

    2016-01-01

    Conspiratorial ideation is the tendency of individuals to believe that events and power relations are secretly manipulated by certain clandestine groups and organisations. Many of these ostensibly explanatory conjectures are non-falsifiable, lacking in evidence or demonstrably false, yet public acceptance remains high. Efforts to convince the general public of the validity of medical and scientific findings can be hampered by such narratives, which can create the impression of doubt or disagreement in areas where the science is well established. Conversely, historical examples of exposed conspiracies do exist and it may be difficult for people to differentiate between reasonable and dubious assertions. In this work, we establish a simple mathematical model for conspiracies involving multiple actors with time, which yields failure probability for any given conspiracy. Parameters for the model are estimated from literature examples of known scandals, and the factors influencing conspiracy success and failure are explored. The model is also used to estimate the likelihood of claims from some commonly-held conspiratorial beliefs; these are namely that the moon-landings were faked, climate-change is a hoax, vaccination is dangerous and that a cure for cancer is being suppressed by vested interests. Simulations of these claims predict that intrinsic failure would be imminent even with the most generous estimates for the secret-keeping ability of active participants-the results of this model suggest that large conspiracies (≥1000 agents) quickly become untenable and prone to failure. The theory presented here might be useful in counteracting the potentially deleterious consequences of bogus and anti-science narratives, and examining the hypothetical conditions under which sustainable conspiracy might be possible.

  10. Assessment of donor heart viability during ex vivo heart perfusion.

    Science.gov (United States)

    White, Christopher W; Ambrose, Emma; Müller, Alison; Li, Yun; Le, Hoa; Hiebert, Brett; Arora, Rakesh; Lee, Trevor W; Dixon, Ian; Tian, Ganghong; Nagendran, Jayan; Hryshko, Larry; Freed, Darren

    2015-10-01

    Ex vivo heart perfusion (EVHP) may facilitate resuscitation of discarded donor hearts and expand the donor pool; however, a reliable means of demonstrating organ viability prior to transplantation is required. Therefore, we sought to identify metabolic and functional parameters that predict myocardial performance during EVHP. To evaluate the parameters over a broad spectrum of organ function, we obtained hearts from 9 normal pigs and 37 donation after circulatory death pigs and perfused them ex vivo. Functional parameters obtained from a left ventricular conductance catheter, oxygen consumption, coronary vascular resistance, and lactate concentration were measured, and linear regression analyses were performed to identify which parameters best correlated with myocardial performance (cardiac index: mL·min(-1)·g(-1)). Functional parameters exhibited excellent correlation with myocardial performance and demonstrated high sensitivity and specificity for identifying hearts at risk of poor post-transplant function (ejection fraction: R(2) = 0.80, sensitivity = 1.00, specificity = 0.85; stroke work: R(2) = 0.76, sensitivity = 1.00, specificity = 0.77; minimum dP/dt: R(2) = 0.74, sensitivity = 1.00, specificity = 0.54; tau: R(2) = 0.51, sensitivity = 1.00, specificity = 0.92), whereas metabolic parameters were limited in their ability to predict myocardial performance (oxygen consumption: R(2) = 0.28; coronary vascular resistance: R(2) = 0.20; lactate concentration: R(2) = 0.02). We concluded that evaluation of functional parameters provides the best assessment of myocardial performance during EVHP, which highlights the need for an EVHP device capable of assessing the donor heart in a physiologic working mode.

  11. Polyphenolic Extracts of Edible Flowers Incorporated onto Atelocollagen Matrices and Their Effect on Cell Viability

    Directory of Open Access Journals (Sweden)

    Jorge López-García

    2013-10-01

    Full Text Available The phenolic extract of chives flowers (Allium schoenoprasum, Liliaceae, introduced Sage (Salvia pratensis, Lamiaceae, European elderberry (Sambucus nigra, Caprifoliaceae and common dandelion (Taraxacum officinale, Asteraceae were characterised by High Performance Liquid Chromatography and incorporated in different concentrations onto atelocollagen thin films. In order to assess the biological impact of these phenolic compounds on cell viability, human immortalised non-tumorigenic keratinocyte cell line was seeded on the thin films and cell proliferation was determined by using an MTT assay. In addition, their antimicrobial activity was estimated by using an agar diffusion test. Data indicated the concomitance between cell viability and concentration of polyphenols. These findings suggest that these phenolic-endowed atelocollagen films might be suitable for tissue engineering applications, on account of the combined activity of polyphenols and collagen.

  12. Rhabdias pseudosphaerocephala infection in Bufo marinus: lung nematodes reduce viability of metamorph cane toads.

    Science.gov (United States)

    Kelehear, C; Webb, J K; Shine, R

    2009-07-01

    Cane toads (Bufo marinus) were introduced to Australia in 1935 and have since spread widely over the continent, generating concern regarding ecological impacts on native predators. Most Australian cane toad populations are infected with lung nematodes Rhabdias pseudosphaerocephala, a parasite endemic to New World (native-range) cane toad populations; presumably introduced to Australia with its toad host. Considering the high intensities and prevalence reached by this parasite in Australian toad populations, and public ardour for developing a control plan for the invasive host species, the lack of experimental studies on this host-parasite system is surprising. To investigate the extent to which this lungworm influences cane toad viability, we experimentally infected metamorph toads (the smallest and presumably most vulnerable terrestrial phase of the anuran life cycle) with the helminth. Infected toads exhibited reduced survival and growth rates, impaired locomotor performance (both speed and endurance), and reduced prey intake. In summary, R. pseudosphaerocephala can substantially reduce the viability of metamorph cane toads.

  13. Reduced pollen viability and achene development in Solidago ×niederederi Khek from Poland

    Directory of Open Access Journals (Sweden)

    Grzegorz Migdałek

    2014-09-01

    Full Text Available Pollen and achene characters of natural interspecific hybrid Solidago ×niederederi Khek were analyzed and compared with putative parental species S. virgaurea L. and S. canadensis L. to estimate the level of disturbances in generative reproduction resulting from its hybrid nature. Pollen viability (stainability of Solidago ×niederederi from one newly discovered locality in NE Poland was evidently reduced to ~65% in both viability tests (acetocarmine and Alexander. The diameter of viable pollen (median 21.11 µm fell between S. canadensis (median 19.52 µm and S. virgaurea (median 23.48 µm. Both parental species produced normally developed achenes with high frequency (~90% whereas in the hybrid, the seed set was dramatically low (6%. The results clearly indicated that sexual reproduction of hybridogenous taxon S. ×niederederi is disturbed, and its potential impact as an invasive species depends mainly on vegetative propagation.

  14. Effect of alcohols on filamentation, growth, viability and biofilm development in Candida albicans.

    Science.gov (United States)

    Chauhan, Nitin M; Shinde, Ravikumar B; Karuppayil, S Mohan

    2013-12-01

    In this study we report the potential of alcohols as morphogenetic regulators in Candida albicans. All the alcohols tested influenced various modes of growth like planktonic as well as biofilm forms. Viability was affected at high concentrations. Among the alcohols, the response of C. albicans to amyl alcohol (pentanol) was noteworthy. Amyl alcohol at a concentration 0.5% which was not inhibitory to growth and viability specifically inhibited morphogenetic switching from yeast to hyphal forms. It also inhibited normal biofilm development favoring yeast dominated biofilms. Based on this study we hypothesize that alcohols produced under anaerobic conditions may not favor biofilm development and support dissemination of yeast cells. Since anaerobic conditions are not found to favor production of quorum sensing molecules like farnesol, the alcohols may play a role in morphogenetic regulation.

  15. Use of fluorescent redox indicators to evaluate cell proliferation and viability

    DEFF Research Database (Denmark)

    Rasmussen, E.S.

    1999-01-01

    The performance of two cell viability test kits based on the use of redox indicators yielding fluorescent products, the AlamarBlue assay and a resazurin-based in vitro toxicology assay kit from Sigma, was compared in the present study. Cultures of human neonatal foreskin fibroblasts were exposed...... to equal concentrations of the two dye solutions in the cell culture media. The fluorescence intensities of the cell culture media obtained in response to cell proliferation with the two dyes showed a pronounced similarity. Both dyes were noncytotoxic to cell cultures with high initial cell densities...... components were tentatively identified as resazurin and resorufin. The AlamarBlue assay has gained wide application as a cell viability indicator that allows continuous monitoring of cell proliferation or cytotoxicity in human and animal cells, bacteria, and fungi, but no studies with the deliberate use...

  16. Conidial vigor vs. viability as predictors of virulence of entomopathogenic fungi.

    Science.gov (United States)

    Faria, Marcos; Lopes, Rogério Biaggioni; Souza, Daniela Aguiar; Wraight, Stephen P

    2015-02-01

    We tested the hypothesis that debilitated conidia exhibiting slow-germination (requiring>16h to germinate) are less virulent than vigorous conidia exhibiting fast germination (requiring⩽16h to germinate). Preparations of Beauveria bassiana s.l. strain CG 1027 with variable ratios of vigorous to debilitated conidia were assayed against third-instar larvae of Spodoptera frugiperda. As the proportion of debilitated conidia in test preparations increased, LC50 expressed in terms of total viable conidia increased, while LC50 expressed solely in terms of vigorous conidia remained constant, indicating that vigorous conidia were responsible for nearly all mortality observed in the assays. Larvae treated with conidia from low-quality batches (with high proportions of debilitated conidia) survived consistently longer than those treated with comparable doses of conidia from high-quality batches. These results confirm our previous hypotheses that inclusion of debilitated conidia in viability assessments can lead to overestimation of the quality (potency) of mycoinsecticide preparations and support our recommendation for use of short incubation periods for assessing viability whenever viability is relied upon as an indicator of product quality. Copyright © 2015. Published by Elsevier Inc.

  17. Testing a dual-fluorescence assay to monitor the viability of filamentous cyanobacteria.

    Science.gov (United States)

    Johnson, Tylor J; Hildreth, Michael B; Gu, Liping; Zhou, Ruanbao; Gibbons, William R

    2015-06-01

    Filamentous cyanobacteria are currently being engineered to produce long-chain organic compounds, including 3rd generation biofuels. Because of their filamentous morphology, standard methods to quantify viability (e.g., plate counts) are not possible. This study investigated a dual-fluorescence assay based upon the LIVE/DEAD® BacLight™ Bacterial Viability Kit to quantify the percent viability of filamentous cyanobacteria using a microplate reader in a high throughput 96-well plate format. The manufacturer's protocol calls for an optical density normalization step to equalize the numbers of viable and non-viable cells used to generate calibration curves. Unfortunately, the isopropanol treatment used to generate non-viable cells released a blue pigment that altered absorbance readings of the non-viable cell solution, resulting in an inaccurate calibration curve. Thus we omitted this optical density normalization step, and carefully divided cell cultures into two equal fractions before the isopropanol treatment. While the resulting calibration curves had relatively high correlation coefficients, their use in various experiments resulted in viability estimates ranging from below 0% to far above 100%. We traced this to the apparent inaccuracy of the propidium iodide (PI) dye that was to stain only non-viable cells. Through further analysis via microplate reader, as well as confocal and wide-field epi-fluorescence microscopy, we observed non-specific binding of PI in viable filamentous cyanobacteria. While PI will not work for filamentous cyanobacteria, it is possible that other fluorochrome dyes could be used to selectively stain non-viable cells. This will be essential in future studies for screening mutants and optimizing photobioreactor system performance for filamentous cyanobacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Lake eutrophication and environmental change: A viability framework for resilience, vulnerability and adaptive capacity

    Science.gov (United States)

    Mathias, Jean-Denis; Rougé, Charles; Deffuant, Guillaume

    2013-04-01

    We present a simple stochastic model of lake eutrophication to demonstrate how the mathematical framework of viability theory fosters operational definitions of resilience, vulnerability and adaptive capacity, and then helps understand which response one should bring to environmental changes. The model represents the phosphorus dynamics, given that high concentrations trigger a regime change from oligotrophic to eutrophic, and causes ecological but also economic losses, for instance from tourism. Phosphorus comes from agricultural inputs upstream of the lake, and we will consider a stochastic input. We consider the system made of both the lake and its upstream region, and explore how to maintain the desirable ecological and economic properties of this system. In the viability framework, we translate these desirable properties into state constraints, then examine how, given the dynamics of the model and the available policy options, the properties can be kept. The set of states for which there exists a policy to keep the properties is called the viability kernel. We extend this framework to both major perturbations and long-term environmental changes. In our model, since the phosphorus inputs and outputs from the lake depend on rainfall, we will focus on extreme rainfall events and long-term changes in the rainfall regime. They can be described as changes in the state of the system, and may displace it outside the viability kernel. Its response can then be described using the concepts of resilience, vulnerability and adaptive capacity. Resilience is the capacity to recover by getting back to the viability kernel where the dynamics keep the system safe, and in this work we assume it to be the first objective of management. Computed for a given trajectory, vulnerability is a measure of the consequence of violating a property. We propose a family of functions from which cost functions and other vulnerability indicators can be derived for any trajectory. There can be

  19. The practice of investment viability appraisal in Akure, Nigeria ...

    African Journals Online (AJOL)

    This paper examines the role played by valuers in choosing the right viability appraisal technique for an investment appraisal. Structured questionnaire was administered on Twenty one (21) registered and practicing Estate Surveying and Valuation firms in Akure out of which fourteen (14) were retrieved and found good for ...

  20. Proof of Economic Viability of Blended Learning Business Models

    Science.gov (United States)

    Druhmann, Carsten; Hohenberg, Gregor

    2014-01-01

    The discussion on economically sustainable business models with respect to information technology is lacking in many aspects of proven approaches. In the following contribution the economic viability is valued based on a procedural model for design and evaluation of e-learning business models in the form of a case study. As a case study object a…

  1. Evaluating the Viability of Mobile Learning to Enhance Management Training

    Science.gov (United States)

    Macdonald, Iain; Chiu, Jason

    2011-01-01

    A qualitative research project was conducted to test the viability of augmenting an e-learning program for workplace learners using mobile content delivered through smart phones. Ten learners taking a six week web-based e-learning course were given smart phones which enabled them to access approximately 70% of the course content, in addition to…

  2. Assessment of viability of microorganisms employing fluorescence techniques

    NARCIS (Netherlands)

    Breeuwer, P.

    1996-01-01


    Viability assessment of microorganisms is relevant for a wide variety of applications in industry, including evaluation of inactivation treatments and quality assessment of starter cultures for beer, wine, and yoghurt production.

    Usually, the ability of microbial cells to

  3. Desiccation-induced changes in viability, lipid peroxidation and ...

    African Journals Online (AJOL)

    Intermediate seeds of Mimusopsis elengi showed obvious membrane lipid peroxidation during desiccation. When the moisture content (MC) decreased from initial 41.8 to 6.1%, seed viability significantly decreased from 100 to 23%, consorted with activity changes of a few anti-oxidative enzymes. The activities of superoxide ...

  4. The effects of storage conditions on the viability of ...

    African Journals Online (AJOL)

    Long-terms recoverability of enteropathogens is necessary for future epidemiological studies to screen stool samples when conditions do not permit immediate processing. The aim of this study was to determine the viability and the recoverability of three enteropathogens bacteria (Yersinia enterocolitica, Vibrio cholerae O: 1 ...

  5. The viability of business data mining in the sports environment ...

    African Journals Online (AJOL)

    Data mining can be viewed as the process of extracting previously unknown information from large databases and utilising this information to make crucial business decisions (Simoudis, 1996: 26). This paper considers the viability of using data mining tools and techniques in sports, particularly with regard to mining the ...

  6. Influence of gamma irradiation on pollen viability, germination ability ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-12-15

    Dec 15, 2009 ... Effects of gamma radiation on vitality and competitive ability of Cucumis pollen. Euphytica, 32: 677-684. Yanmaz R, Ellialtıoglu S, Taner KY (1999). The effects of gamma irradiation on pollen viability and haploid plant formation in snake cucumber (Cucumis melo L. var. flexuosus Naud.). Acta Hort. 492:.

  7. Low-level waste vitrification contact maintenance viability study

    Energy Technology Data Exchange (ETDEWEB)

    Leach, C.E., Westinghouse Hanford

    1996-07-12

    This study investigates the economic viability of contact maintenance in the Low-Level Waste Vitrification Facility, which is part of the Hanford Site Tank Waste Remediation System. This document was prepared by Flour Daniel, Inc., and transmitted to Westinghouse Hanford Company in September 1995.

  8. Pollen diversity, viability and floral structure of some Musa genotypes

    African Journals Online (AJOL)

    This experiment was designed to study the floral structure, pollen morphology and the potential pollen viability of five Musa genotypes obtained from the Musa field ... Three different types of pollen were encountered viz, big, moderate and small pollens with corresponding big, moderate and small apertures and pores.

  9. Viability of bull semen extended with commercial semen extender ...

    African Journals Online (AJOL)

    Andrea Raseona

    Abstract. The aim of this study was to evaluate the viability of bull spermatozoa diluted with commercial semen extender and two culture media stored at controlled room temperature (24 °C) for 72 hours. Two Nguni bulls were used for semen collection with the aid of an electro-ejaculator. After macroscopic evaluation ...

  10. The economic and social viability of Tanzanian Wildlife Management Areas

    DEFF Research Database (Denmark)

    Homewood, Katherine; Bluwstein, Jevgeniy; Lund, Jens Friis

    This policy brief contributes to assessing the economic and social viability of Tanzania’s Wildlife Management Areas (WMAs) through preliminary findings by the ‘Poverty and ecosystem Impacts of Tanzania’s Wildlife Management Areas’ (PIMA) project, focusing on benefits, costs, and their distribution...

  11. Effect of pretreatments on seed viability during fruit development of ...

    African Journals Online (AJOL)

    Studies to identify the stage at which developing fruits of Irvingia gabonensis (var. excelsa and var. gabonensis), picked from standing trees and/or forest floors, attain maximum viability and germinability were conducted in two harvesting seasons in 2000 and 2001. Some pretreatment methods were used as a means of ...

  12. Viability of bull semen extended with commercial semen extender ...

    African Journals Online (AJOL)

    After macroscopic evaluation, semen was pooled and aliquoted randomly into Triladyl, modified Ham's F10, and TCM-199 culture media, and then stored at 24 °C. Sperm motility parameters, morphology, and viability were analysed with computer aided sperm analysis (CASA) after 0, 24, 48 and 72 hours. The study was ...

  13. Potential carbon credit and community expectations towards viability ...

    African Journals Online (AJOL)

    The paper presents results of the potential carbon credit and community expectations towards viability of REDD+ projects in Ugalla- Masito ecosystem using a case of Ilagala and Karago villages whereby REDD+ is being piloted. Various data collection methods were employed and these included focused group discussion, ...

  14. Research Note on viability of herbicide and Hormone - treated ...

    African Journals Online (AJOL)

    The responses and viability of acid scarified seeds of four tropical weeds to gibberellic acid and seven herbicides including Galex, Gramoxone, 2 - 4 D, Atrazine, Simazine, Roundup and Primextra in the Laboratory were investigated. The weeds used are Cassia occidentalis, Cassia obtusifolia Cassia hirtusa and ...

  15. Economic Viability of Deficit Irrigation in the Western US

    Science.gov (United States)

    In many arid regions of the world, population growth, groundwater depletion, and uncertain supplies have caused agricultural water to become increasingly scarce. Deficit irrigation (DI) provides a potential response to water scarcity, but no consensus exists on its economic viability. In this pape...

  16. Port viability for choice making among shipping companies in West ...

    African Journals Online (AJOL)

    In this paper, the study of port viability for choice making among shipping companies in West Africa sub-region trade route was conducted. Discriminant analysis was used to ascertain the consistency of the attributes of ports that establish their overall attractiveness to the carriers. The critical valued port attributes deduced ...

  17. The effects of storage conditions on the viability of ...

    African Journals Online (AJOL)

    SARAKA DANIEL

    2015-01-07

    Jan 7, 2015 ... Long-terms recoverability of enteropathogens is necessary for future epidemiological studies to screen stool samples when conditions do not permit immediate processing. The aim of this study was to determine the viability and the recoverability of three enteropathogens bacteria (Yersinia enterocolitica,.

  18. Relationships between cock semen viability and the fertility of ...

    African Journals Online (AJOL)

    CUT User

    Semen was collected from each cock following 5ASM, evaluated for semen viability and 0.05 mL diluted semen used to inseminate five hens per breed, in each experimental group. Significant differences in ejaculation rates and semen quality and quantity were recorded in the four breeds of cocks - with the HP cocks of the ...

  19. Optimizing cell viability in droplet-based cell deposition

    NARCIS (Netherlands)

    Hendriks, Jan; Willem Visser, Claas; Henke, Sieger; Leijten, Jeroen; Saris, Daniël B F|info:eu-repo/dai/nl/241604443; Sun, Chao; Lohse, Detlef; Karperien, Marcel

    2015-01-01

    Biofabrication commonly involves the use of liquid droplets to transport cells to the printed structure. However, the viability of the cells after impact is poorly controlled and understood, hampering applications including cell spraying, inkjet bioprinting, and laser-assisted cell transfer. Here,

  20. Interactions between Plant Extracts and Cell Viability Indicators ...

    African Journals Online (AJOL)

    Interactions between Plant Extracts and Cell Viability. Indicators during Cytotoxicity Testing: Implications for. Ethnopharmacological Studies. Sze Mun Chan1, Kong Soo Khoo2 and Nam Weng Sit1*. 1Department of Biomedical Science, 2Department of Chemical Science, Faculty of Science, Universiti Tunku Abdul Rahman,.

  1. Dormancy, activation and viability of Rhizopus oligosporus sporangiospores

    NARCIS (Netherlands)

    Thanh, N.V.; Nout, M.J.R.

    2004-01-01

    Interruption of dormancy to improve viability of Rhizopus oligosporus sporangiospores is crucial for the application of stored starter cultures for fungal (tempe) production. We aimed to assess the extent of dormancy and factors that could result in activation. Whereas heat treatments were

  2. Influence of gamma irradiation on pollen viability, germination ability ...

    African Journals Online (AJOL)

    July 9th, 11th, 15th, 21st and 28th) and pollen age (0th and 1st days) on the pollen viability, germination ability and fruit and seed-set were investigated in pumpkin (Cucurbita moschata Duchesne ex Poir.) and winter squash (Cucurbita maxima ...

  3. N-terminal arm of orchardgrass Hsp17.2 (DgHsp17.2) is essential for both in vitro chaperone activity and in vivo thermotolerance in yeast.

    Science.gov (United States)

    Cha, Joon-Yung; Lee, Sang-Hoon; Seo, Kyung Hye; Choi, Young Jin; Cheong, Mi Sun; Son, Daeyoung

    2016-02-01

    Small heat shock proteins are well-known to function as chaperone in the protection of proteins and subcellular structures against stress-induced denaturation in many cell compartments. Irrespective of such general functional assignment, a proof of function in a living organism is missing. Here, we used heat-induced orchardgrass small Hsp17.2 (DgHsp17.2). Its function in in vitro chaperone properties has shown in protecting the model substrate, malate dehydrogenase (MDH) and citrate synthase (CS). Overexpression of DgHsp17.2 triggering strong chaperone activity enhanced in vivo thermotolerance of yeast cells. To identify the functional domain on DgHsp17.2 and correlationship between in vitro chaperone property and in vivo thermotolerance, we generated truncation mutants of DgHsp17.2 and showed essentiality of the N-terminal arm of DgHsp17.2 for the chaperone function. In addition, beyond for acquisition of thermotolerance irrespective of sequences are diverse among the small Hsps. However, any truncation mutants of DgHsp17.2 did not exhibit strong interaction with orchardgrass heat shock protein 70 (DgHsp70) different from mature DgHsp17.2, indicating that full-length DgHsp17.2 is necessary for cooperating with Hsp70 protein. Our study indicates that the N-terminal arm of DgHsp17.2 is an important region for chaperone activity and thermotolerance. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. The effect of ketorolac tromethamine, methylprednisolone, and platelet-rich plasma on human chondrocyte and tenocyte viability.

    Science.gov (United States)

    Beitzel, Knut; McCarthy, Mary Beth; Cote, Mark P; Apostolakos, John; Russell, Ryan P; Bradley, James; ElAttrache, Neal S; Romeo, Antony A; Arciero, Robert A; Mazzocca, Augustus D

    2013-07-01

    The purpose of this study was to evaluate the effect on cell viability of the isolated and combined use of allogeneic platelet-rich plasma (PRP) and ketorolac tromethamine on human chondrocytes and tenocytes in a highly controlled in vitro environment. PRP was produced from 8 subjects. Human chondrocytes (Lonza, Hopkinton, MA) and tenocytes isolated from samples of the long head of the biceps tendons were treated in culture with PRP, ketorolac tromethamine, and methylprednisolone, both alone and in combination. Control samples were treated in media containing 2% or 10% fetal bovine serum (FBS). Cells were exposed for 1 hour. Luminescence assays were obtained to examine cell viability after 24 hours and long-term effects on cell viability after 120 hours. Radioactive thymidine assay was used to measure proliferation after 120 hours. For chondrocytes, cell viability (120 hours) increased significantly with the treatment of PRP alone (43,949 ± 28,104 cells; P investigation into alternative treatment options such as combinations of PRP and ketorolac tromethamine. In vitro evaluation of their effect on cell viability might build a basis for further translational research and clinical application. Copyright © 2013 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.

  5. Constant, cycling, hot and cold thermal environments: strong effects on mean viability but not on genetic estimates.

    Science.gov (United States)

    Ketola, T; Kellermann, V; Kristensen, T N; Loeschcke, V

    2012-06-01

    It has frequently been suggested that trait heritabilities are environmentally sensitive, and there are genetic trade-offs between tolerating different environments such as hot and cold or constant and fluctuating temperatures. Future climate predictions suggest an increase in both temperatures and their fluctuations. How species will respond to these changes is uncertain, particularly as there is a lack of studies which compare genetic performances in constant vs. fluctuating environments. In this study, we used a nested full-sib/half-sib breeding design to examine how the genetic variances and heritabilities of egg-to-adult viability differ at high and low temperatures with and without daily fluctuations in temperatures using Drosophila melanogaster as a model organism. Although egg-to-adult viability was clearly sensitive to developmental temperatures, heritabilities were not particularly sensitive to developmental temperatures. Moreover, we found that egg-to-adult viabilities at different developmental temperatures were positively correlated, suggesting a common genetic background for egg-to-adult viability at different temperatures. Finding both a uniform genetic background coupled with rather low heritabilities insensitive to temperatures, our results suggest evolutionary responses are unlikely to be limited by temperature effects on genetic parameters or negative genetic correlations, but by the direct effects of stressful temperatures on egg-to-adult viability accompanied with low heritabilities. © 2012 The Authors. Journal of Evolutionary Biology © 2012 European Society For Evolutionary Biology.

  6. Caffeine inhibits the viability and osteogenic differentiation of rat bone marrow-derived mesenchymal stromal cells.

    Science.gov (United States)

    Zhou, Y; Guan, X X; Zhu, Z L; Guo, J; Huang, Y C; Hou, W W; Yu, H Y

    2010-12-01

    Caffeine is consumed extensively in Europe and North America. As a risk factor for osteoporosis, epidemiological studies have observed that caffeine can decrease bone mineral density, adversely affect calcium absorption and increase the risk of bone fracture. However, the exact mechanisms have not been fully investigated. Here, we examined the effects of caffeine on the viability and osteogenesis of rat bone marrow-derived mesenchymal stromal cells (rBMSCs). Cell viability, apoptosis and necrosis were quantified using thymidine incorporation and flow cytometry. Sequential gene expressions in osteogenic process were measured by real-time PCR. cAMP, alkaline phosphatase and osteocalcin were assessed by immunoassay, spectrophotometry and radioimmunoassay, respectively. Mineralization was determined by calcium deposition. After treating BMSCs with high caffeine concentrations (0.1-1mM), their viability decreased in a concentration-dependent manner. This cell death was primarily due to necrosis and, to a small extent, apoptosis. Genes and protein sequentially expressed in osteogenesis, including Cbfa1/Runx2, collagen I, alkaline phosphatase and its protein, were significantly downregulated except for osteocalcin and its protein. Moreover, caffeine inhibited calcium deposition in a concentration- and time-dependent manner, but increased intracellular cAMP in a concentration-dependent manner. By suppressing the commitment of BMSCs to the osteogenic lineage and selectively inhibiting gene expression, caffeine downregulated some important events in osteogenesis and ultimately affected bone mass.

  7. Enhancing the viability of Lactobacillus rhamnosus GG after spray drying and during storage.

    Science.gov (United States)

    Broeckx, Géraldine; Vandenheuvel, Dieter; Henkens, Tim; Kiekens, Shari; van den Broek, Marianne F L; Lebeer, Sarah; Kiekens, Filip

    2017-10-03

    Increasing knowledge about the human microbiome has led to a growing awareness of the potential of applying probiotics to improve our health. The pharmaceutical industry shows an emerging interest in pharmaceutical formulations containing these beneficial microbes, the so-called pharmabiotics. An important manufacturing step is the drying of the probiotics, as this can increase the stability and shelf life of the finished pharmabiotic product. Unfortunately, drying also puts stress on microbial cells, thus causing a decrease in viability. We aimed to examine the effect of different drying media and protective excipients on the viability of the prototype probiotic strain Lactobacillus rhamnosus GG after spray drying and during subsequent storage for 28 weeks. The presence of phosphates in the drying medium showed to have a superior protective effect, especially during long-term storage at room temperature. Addition of lactose or trehalose resulted in significantly improved survival rates after drying as well as during long-term storage for the tested excipients. Both disaccharides are characterized by a high glass transition temperature. Maltodextrin showed less protective capacities compared to lactose and trehalose in all tested conditions. The usage of mannitol or dextran resulted in sticky powders and low yields, so further testing was not possible. In addition to optimizing the viability, future research will also explore the functionality of cellular probiotic components after spray drying in order to safeguard the probiotic activity of the formulated pharmabiotics. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Lowering the concentration affects the migration and viability of intracerebroventricular-delivered human mesenchymal stem cells.

    Science.gov (United States)

    Kim, Hyeong Seop; Lee, Na Kyung; Yoo, Dongkyeom; Lee, Jeongmin; Choi, Soo Jin; Oh, Wonil; Chang, Jong Wook; Na, Duk L

    2017-11-04

    Due to their widely known therapeutic benefits, mesenchymal stem cells have been proposed as a novel treatment option for a wide range of diseases including Alzheimer's disease. To maximize these benefits, critical factors such as delivery route, cell viability, and cell migration must be accounted for. Out of the various delivery routes to the brain, the intracerebroventricular (ICV) route stands out due to the widespread distribution that can occur via cerebrospinal fluid flow. The major objective of this present study was to observe how altering cell concentration influences the migration and viability of human umbilical cord blood derived-mesenchymal stem cells (hUCB-MSCs), delivered via ICV injection, in the brains of wild-type (WT) mice. C3H/C57 WT mice were divided into three groups and were injected with 1 × 10(5) hUCB-MSCs suspended in varying volumes: high (3 μl), middle (5 μl), and low (7 μl) concentrations, respectively. Lowering the concentration increased the migratory capabilities and elevated the viability of hUCB-MSCs. These results suggest that cell concentration can affect the physiological state of hUCB-MSCs, and thus the extent of therapeutic efficacy that can be achieved. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  9. Resveratrol Inhibits Alpha-Melanocyte-Stimulating Hormone Signaling, Viability, and Invasiveness in Melanoma Cells

    Directory of Open Access Journals (Sweden)

    Yu-Jen Chen

    2013-01-01

    Full Text Available Melanoma is a malignancy with high potential to invasion and treatment resistance. The α-melanocyte-stimulating hormone (α-MSH signal transduction involving Wnt/β-catenin, c-Kit, and microphthalmia-associated transcription factor (MITF, a known pathway to produce melanin, has been demonstrated as one of cancer stem cell characteristics. This study was aimed to examine the effect of resveratrol, an abundant ingredient of grape and medicinal plants, on α-MSH signaling, viability, and invasiveness in melanoma cells. By α-MSH treatment, the melanin production in B16 melanoma cells was augmented as a validation for activation of α-MSH signaling. The upregulated expression of α-MSH signaling-related molecules β-catenin, c-Kit, and MITF was suppressed by resveratrol and/or STI571 treatment. Nuclear translocation of MITF, a hallmark of α-MSH signaling activation, was inhibited by combined treatment of resveratrol and STI571. At effective concentration, resveratrol and/or STI571 inhibited cell viability and α-MSH-activated matrix metalloproteinase- (MMP-9 expression and invasion capacity of B16 melanoma cells. In conclusion, resveratrol enhances STI571 effect on suppressing the α-MSH signaling, viability, and invasiveness in melanoma cells. It implicates that resveratrol may have potential to modulate the cancer stem cell characteristics of melanoma.

  10. Postmortem morphology and viability of human Peyer's patches in distal ileum: a technical note.

    Science.gov (United States)

    Solarino, Biagio; Di Vella, Giancarlo; Magrone, Thea; Jirillo, Felicita; Tafaro, Angela; Piscitelli, Domenico; Casiello, Michela; Amati, Luigi; Jirillo, Emilio; Tattoli, Lucia

    2009-01-01

    The intestinal mucosa contains a highly specialized immune system which plays a central role in the induction of immune reactions. In the small bowel, Gut-Associated Lymphoid Tissue (GALT) is organized in lymphoid aggregates which are known as Peyer's Patches (PP). Even though human PP involvement in systemic immunity has been described, little is known about their anatomy and morphology and viability. The aim of this study was to examine PP according to their macroscopic anatomy, distribution and cell viability after death. Specimens from the distal ileum were obtained from 72 serial autopsy cases: PP were identified and, parts of them were analyzed for histological examination. Moreover, viability of recovered PP cells was assessed by the trypan blue exclusion test. Most of the PP (90%) were situated on the antimesenteric border of ileum, and the greatest density of PP occurred in the most distal segment. The number of PP varied with age, with the maximum number observed in 21- to 30-years old cadavers. Histological examination showed their remarkable architectural preservation at different post-mortem intervals (PMI), while the mucosal surface underwent autolysis. In 56% of cases PP cells were still viable, especially at PMI < 24 hours after death. These data confirm that human PP are still well preserved in a remarkable percentage of cadavers also several hours after death, and their availability may be helpful in various fields of research.

  11. Changes in the Number, Viability, and Amino-acid-incorporating Activity of Rhizobium Bacteroids during Lupin Nodule Development

    Science.gov (United States)

    Sutton, William D.; Jepsen, Neil M.; Shaw, Brian D.

    1977-01-01

    Between days 10 and 21 after inoculation of Lupinus angustifolius seedlings with Rhizobium NZP 2257, the average nodule fresh weight increased 3-fold and the number of bacteroids per nodule increased more than 10-fold. The viability of Rhizobium bacteroids, as judged by their ability to form colonies on yeast-extract agar, declined from about 10% on days 10 and 11 after inoculation to about 0.3% on days 14 to 25. Bacteroid viability was highly sensitive to osmolarity. At optimal pH and K and Mg ion concentrations, the incorporation of 14C-glycine into isolated bacteroids was also very sensitive to osmolarity, and fell in parallel with bacteroid viability during nodule development. We suggest that at least two processes contribute to bacteroid nonviability: a reversible change in the cell wall structure occurring between days 10 and 14 after inoculation, and a subsequent irreversible change. PMID:16659929

  12. MAINTENANCE OF VIABILITY AND TRANSPORT FUNCTION AFTER PRESERVATION OF ISOLATED RAT HEPATOCYTES IN VARIOUS SIMPLIFIED UNIVERSITY-OF-WISCONSIN SOLUTIONS

    NARCIS (Netherlands)

    SANDKER, GW; WEERT, B; KUIPERS, W; SLOOFF, MJH; MEIJER, DKF; GROOTHUIS, GMM; Merema, M.T.

    1993-01-01

    Rat hepatocytes were preserved for 24hr with high recovery and good maintenance of viability and transport function both in University of Wisconsin (UW) solution and in various simplified UW solutions. Cell quality is somewhat affected after 48 hr of preservation in both the original UW solution and

  13. Fetal viability as a threshold to personhood. A legal analysis.

    Science.gov (United States)

    Peterfy, A

    1995-12-01

    This essay opens with an examination of US laws concerning fetal viability as they apply to induced abortion, to a mother's right to refuse medical treatment necessary to save the life of a fetus, and to the rights to file suit for the wrongful death of unborn children. The history of abortion policies in the US is traced from the common law period of the early 19th century to the restrictive post-Civil War laws and the decisions of the Supreme Court in Roe vs. Wade, which upheld the constitutionality of previability abortions; Webster vs. Reproductive Health Services, in which the Court assigned viability to the 20th week of pregnancy and acknowledged that States could have a compelling previability interest in the fetus; and the Casey decision, which provided tolerance for limits on the availability of abortion before viability as long as the limits did not create an "undue burden" on the woman seeking the abortion. Courts dealing with the issue of compelling a mother to undergo medical treatment to save her fetus have been inconsistent as they balanced the state's interest in the fetus against the mother's rights to privacy. Judges have tended to err on the side of forcing the medical interventions, but the most recent trend is against this sort of judgement. In these cases, fetal viability has also served as a dividing line. The inconsistency of the legal system is illustrated by the fact that, whereas the fetus now has a legal existence, wrongful death actions entered on behalf of a nonviable fetus have often been denied although courts have been more willing to extend protection to fetuses in wrongful death tort cases than in abortion or medical intervention cases. Criminal law has a unique set of rules for dealing with fetuses as some states have broadened their definitions of "homicide" to include fetuses, even nonviable fetuses. Courts, however, are reluctant to enlarge criminal statutes on their own. While the central position given to the role of

  14. VIABILITY OF THE PROBIOTIC BACTERIA L. ACIDOPHILUS IN DAIRY PRODUCTS

    Directory of Open Access Journals (Sweden)

    Janka Koreňová

    2011-12-01

    Full Text Available A number of health benefits have been claimed for probiotic bacteria such as Lactobacillus acidophilus. Because of the potential health benefits, these organisms are increasingly incorporated into dairy foods. Viability of probiotic bacteria is important in order to provide health benefits. However, many studies have shown low viability of probiotics in market preparations. This study cover selective enumeration and survival of probiotic bacteria L. acidophilus in some dairy drinks. L. acidophilus was found in the range from 106 to 107 CFU.g-1 in five types of fermented milk products containing probiotic cultures. Two investigated products were up to standard according to Regulation of Ministry of Agriculture and Ministry of Health of Slovak Republic.doi: 10.5219/147

  15. Myocardial Viability: From Proof of Concept to Clinical Practice

    Directory of Open Access Journals (Sweden)

    Aditya Bhat

    2016-01-01

    Full Text Available Ischaemic left ventricular (LV dysfunction can arise from myocardial stunning, hibernation, or necrosis. Imaging modalities have become front-line methods in the assessment of viable myocardial tissue, with the aim to stratify patients into optimal treatment pathways. Initial studies, although favorable, lacked sufficient power and sample size to provide conclusive outcomes of viability assessment. Recent trials, including the STICH and HEART studies, have failed to confer prognostic benefits of revascularisation therapy over standard medical management in ischaemic cardiomyopathy. In lieu of these recent findings, assessment of myocardial viability therefore should not be the sole factor for therapy choice. Optimization of medical therapy is paramount, and physicians should feel comfortable in deferring coronary revascularisation in patients with coronary artery disease with reduced LV systolic function. Newer trials are currently underway and will hopefully provide a more complete understanding of the pathos and management of ischaemic cardiomyopathy.

  16. Traffic networks as information systems a viability approach

    CERN Document Server

    Aubin, Jean-Pierre

    2017-01-01

    This authored monograph covers a viability to approach to traffic management by advising to vehicles circulated on the network the velocity they should follow for satisfying global traffic conditions;. It presents an investigation of three structural innovations: The objective is to broadcast at each instant and at each position the advised celerity to vehicles, which could be read by auxiliary speedometers or used by cruise control devices. Namely, 1. Construct regulation feedback providing at each time and position advised velocities (celerities) for minimizing congestion or other requirements. 2. Taking into account traffic constraints of different type, the first one being to remain on the roads, to stop at junctions, etc. 3. Use information provided by the probe vehicles equipped with GPS to the traffic regulator; 4. Use other global traffic measures of vehicles provided by different types of sensors; These results are based on convex analysis, intertemporal optimization and viability theory as mathemati...

  17. Care at the edge of viability: medical and ethical issues.

    Science.gov (United States)

    Haward, Marlyse F; Kirshenbaum, Nancy W; Campbell, Deborah E

    2011-09-01

    Decision-making for extremely immature preterm infants at the margins of viability is ethically, professionally, and emotionally complicated. A standard for prenatal consultation should be developed incorporating assessment of parental decision-making preferences and styles, a communication process involving a reciprocal exchange of information, and effective strategies for decisional deliberation, guided by and consistent with parental moral framework. Professional caregivers providing perinatal consultations or end-of-life counseling for extremely preterm infants should be sensitive to these issues and be taught flexibility in counseling techniques adhering to consistent guidelines. Emphasis must shift away from physician beliefs and behaviors about the boundaries of viability. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. Characteristics associated with regional health information organization viability.

    Science.gov (United States)

    Adler-Milstein, Julia; Landefeld, John; Jha, Ashish K

    2010-01-01

    Regional Health Information Organizations (RHIOs) will likely play a key role in our nation's effort to catalyze health information exchange. Yet we know little about why some efforts succeed while others fail. We sought to identify factors associated with RHIO viability. Using data from a national survey of RHIOs that we conducted in mid-2008, we examined factors associated with becoming operational and factors associated with financial viability. We used multivariate logistic regression models to identify unique predictors. We classified RHIOs actively facilitating data exchange as operational and measured financial viability as the percent of operating costs covered by revenue from participants in data exchange (0-24%, 25-74%, 75-100%). Predictors included breadth of participants, breadth of data exchanged, whether the RHIO focused on a specific population, whether RHIO participants had a history of collaborating, and sources of revenue during the planning phase. Exchanging a narrow set of data and involving a broad group of stakeholders were independently associated with a higher likelihood of being operational. Involving hospitals and ambulatory physicians, and securing early funding from participants were associated with a higher likelihood of financial viability, while early grant funding seemed to diminish the likelihood. Finding ways to help RHIOs become operational and self-sustaining will bolster the current approach to nationwide health information exchange. Our work suggests that convening a broad coalition of stakeholders to focus on a narrow set of data is an important step in helping RHIOs become operational. Convincing stakeholders to financially commit early in the process may help RHIOs become self-sustaining.

  19. Effect of Allium sativum (garlic) methanol extract on viability and ...

    African Journals Online (AJOL)

    Effect of Allium sativum (garlic) methanol extract on viability and apoptosis of human leukemic cell lines. ... bromide (MTT) assay at concentrations of 3.125, 6.25, 12.5, 25, 50, 100, 200, 400 and 800 ug/mL of Allium sativum extract following 48-h treatment on U-937, Jurkat Clone E6-1 and K-562 cell lines. The mode of cell ...

  20. Viability and Indication of Pathogenic Microbes in the Environment,

    Science.gov (United States)

    viability of parasitic microbes can be clarified corrently only with consideration of the interaction of the organism with the environment and...adaptation to it. According to this school of thought, the stability of a causative agent in the environment is determined by the specific mechanism through...mechanism of transfer of the contaminating principle, the shorter the period during which the parasitic microbe is in the environment - i.e., the

  1. Effect of Isolation Techniques on Viability of Bovine Blood Neutrophils

    Directory of Open Access Journals (Sweden)

    P. Sláma

    2006-01-01

    Full Text Available The effect of selected isolation methods on the viability of neutrophil granulocytes (neutrophils from the blood of healthy Holstein x Bohemian Red Pied crossbred heifers was evaluated. Two methods of neutrophil isolation were used: a neutrophil isolation on the basis of hypotonic erythrocyte lysis (in two variants: after the erythrocyte lysis proper, the cells were centrifuged at either 200 g or 1000 g, and b neutrophil isolation with FACS Lysing Solution as the lysing agent. The viability of the isolated neutrophils was evaluated on the basis of apoptosis and necrosis. The results obtained with flow cytometry (FCM suggest that, from the isolation techniques used, the method based on FACS Lysing Solution impaired the neutrophil viability least. After the application of this method, 5.36 ± 2.15% of neutrophils were apoptotic and 0.51 ± 0.12% were necrotic. In contrast, when the hypotonic erythrocyte lysis was used, the proportion of apoptotic neutrophils amounted to 42.14 ± 7.12% and 49.00 ± 14.70%, respectively, and 41.12 ± 5.55% and 36.91 ± 24.38% respectively of necrotic neutrophils (P < 0.01. This was also confirmed by the light microscopy. After the isolation with FASC Lysing Solution, 1.92 ± 1.74% of neutrophils were apoptotic and 1.05 ± 0.76% were necrotic, as distinct from after the hypotonic erythrocyte lysis where 9.43 ± 3.69% of neutrophils were apoptotic and 12.67 ± 4.74% of necrotic after centrifugation at 200 g, while 12.60 ± 4.35 were apoptotic and 14.96 ± 12.64% were necrotic after centrifugation at 1000 g. It follows from the above-mentioned data that hypotonic lysis is not a suitable method for the isolation of neutrophils, as the method itself markedly affects cell viability.

  2. Effects of Fluid Shear Stress on Cancer Stem Cell Viability

    Science.gov (United States)

    Sunday, Brittney; Triantafillu, Ursula; Domier, Ria; Kim, Yonghyun

    2014-11-01

    Cancer stem cells (CSCs), which are believed to be the source of tumor formation, are exposed to fluid shear stress as a result of blood flow within the blood vessels. It was theorized that CSCs would be less susceptible to cell death than non-CSCs after both types of cell were exposed to a fluid shear stress, and that higher levels of fluid shear stress would result in lower levels of cell viability for both cell types. To test this hypothesis, U87 glioblastoma cells were cultured adherently (containing smaller populations of CSCs) and spherically (containing larger populations of CSCs). They were exposed to fluid shear stress in a simulated blood flow through a 125-micrometer diameter polyetheretherketone (PEEK) tubing using a syringe pump. After exposure, cell viability data was collected using a BioRad TC20 Automated Cell Counter. Each cell type was tested at three physiological shear stress values: 5, 20, and 60 dynes per centimeter squared. In general, it was found that the CSC-enriched U87 sphere cells had higher cell viability than the CSC-depleted U87 adherent cancer cells. Interestingly, it was also observed that the cell viability was not negatively affected by the higher fluid shear stress values in the tested range. In future follow-up studies, higher shear stresses will be tested. Furthermore, CSCs from different tumor origins (e.g. breast tumor, prostate tumor) will be tested to determine cell-specific shear sensitivity. National Science Foundation Grant #1358991 supported the first author as an REU student.

  3. MODERN TECHNIQUES OF CERVICAL INSTRUMENTATION IN IMMATURE SKELETON: VIABILITY ASSESSMENT

    OpenAIRE

    Aires, Ayrana Soares; Silva, Luís Eduardo Carelli Teixeira da; Barros, Alderico Girão Campos de; Azevedo, Gustavo Borges Laurindo de; Naves, Cleiton Dias

    2017-01-01

    ABSTRACT Objective: This study describes the use of materials for modern cervical instrumentation, evaluating its viability in children and adolescents, and the techniques used in different cases. The efficacy of the techniques was analyzed through improvement of pain, maintenance of cervical range of motion, recovery of craniocervical stability, bone consolidation, and spinal stenosis in the postoperative follow-up. Method: Retrospective study of the clinical and radiological parameters of 2...

  4. Comparison of tissue viability imaging and colorimetry: skin blanching.

    Science.gov (United States)

    Zhai, Hongbo; Chan, Heidi P; Farahmand, Sara; Nilsson, Gert E; Maibach, Howard I

    2009-02-01

    Operator-independent assessment of skin blanching is important in the development and evaluation of topically applied steroids. Spectroscopic instruments based on hand-held probes, however, include elements of operator dependence such as difference in applied pressure and probe misalignment, while laser Doppler-based methods are better suited for demonstration of skin vasodilatation than for vasoconstriction. To demonstrate the potential of the emerging technology of Tissue Viability Imaging (TiVi) in the objective and operator-independent assessment of skin blanching. The WheelsBridge TiVi600 Tissue Viability Imager was used for quantification of human skin blanching with the Minolta chromameter CR 200 as an independent colorimeter reference method. Desoximetasone gel 0.05% was applied topically on the volar side of the forearm under occlusion for 6 h in four healthy adults. In a separate study, the induction of blanching in the occlusion phase was mapped using a transparent occlusion cover. The relative uncertainty in the blanching estimate produced by the Tissue Viability Imager was about 5% and similar to that of the chromameter operated by a single user and taking the a(*) parameter as a measure of blanching. Estimation of skin blanching could also be performed in the presence of a transient paradoxical erythema, using the integrated TiVi software. The successive induction of skin blanching during the occlusion phase could readily be mapped by the Tissue Viability Imager. TiVi seems to be suitable for operator-independent and remote mapping of human skin blanching, eliminating the main disadvantages of methods based on hand-held probes.

  5. Criteria for viability assessment of discarded human donor livers during ex vivo normothermic machine perfusion.

    Directory of Open Access Journals (Sweden)

    Michael E Sutton

    Full Text Available Although normothermic machine perfusion of donor livers may allow assessment of graft viability prior to transplantation, there are currently no data on what would be a good parameter of graft viability. To determine whether bile production is a suitable biomarker that can be used to discriminate viable from non-viable livers we have studied functional performance as well as biochemical and histological evidence of hepatobiliary injury during ex vivo normothermic machine perfusion of human donor livers. After a median duration of cold storage of 6.5 h, twelve extended criteria human donor livers that were declined for transplantation were ex vivo perfused for 6 h at 37 °C with an oxygenated solution based on red blood cells and plasma, using pressure controlled pulsatile perfusion of the hepatic artery and continuous portal perfusion. During perfusion, two patterns of bile flow were identified: (1 steadily increasing bile production, resulting in a cumulative output of ≥ 30 g after 6 h (high bile output group, and (2 a cumulative bile production <20 g in 6 h (low bile output group. Concentrations of transaminases and potassium in the perfusion fluid were significantly higher in the low bile output group, compared to the high bile output group. Biliary concentrations of bilirubin and bicarbonate were respectively 4 times and 2 times higher in the high bile output group. Livers in the low bile output group displayed more signs of hepatic necrosis and venous congestion, compared to the high bile output group. In conclusion, bile production could be an easily assessable biomarker of hepatic viability during ex vivo machine perfusion of human donor livers. It could potentially be used to identify extended criteria livers that are suitable for transplantation. These ex vivo findings need to be confirmed in a transplant experiment or a clinical trial.

  6. Ca-Lignosulphonate and sclerotial viability of Sclerotinia sclerotiorum

    Directory of Open Access Journals (Sweden)

    MATTEO MONTANARI

    2012-01-01

    Full Text Available Lignosulphonates, low cost by-products of the pulping process, have shown suppressive effects against some diseases caused by soil-borne pathogens. In this study, the effect of 1.5% v/v calcium lignosulphonate (Ca-Ls amendment to two commercial potting mixes (peat + coconut fibres; PC; and municipal compost + peat + pumice; MCPP on the viability of Sclerotinia sclerotiorum sclerotia was investigated. Sclerotia were buried in the Ca-Ls amended substrates for 30 days. Non-amended PC and MCPP, sterile sand and sterile PC with and without Ca-Ls were used as controls. The viability of sclerotia recovered from PC and MCPP amended with Ca-Ls was reduced by 50 and 42% respectively compared to control treatments. Ca-Ls amendment decreased sclerotial viability by enhancing the activity of the indigenous mycoparasitic fungi, Fusarium oxysporum, Mucor spp. and Trichoderma spp. The biocontrol ability of Ca-Ls against sclerotia was due to the stimulation of microbial activity and is, therefore, strictly dependent on the microbial composition of the substrate.

  7. Viability studies of optically trapped T-cells

    Science.gov (United States)

    McAlinden, Niall; Glass, David G.; Millington, Owain; Wright, Amanda J.

    2011-10-01

    We present a viability study of optically trapped live T cell hybridomas. T cells form an important part of the adaptive immune response system which is responsible for fighting particular pathogens or diseases. The cells of interest were directly trapped by a laser operating at a wavelength of 1064 nm and their viability measured as a function of time. Cell death was monitored using an inverted fluorescent microscope to observe the uptake by the cell of the fluorescent dye propidium iodide. Studies were undertaken at various laser powers and beam profiles. There is a growing interest in optically trapping immune cells and this is the first study that investigates the viability of a T cell when trapped using a conventional optical trapping system. In such experiments it is crucial that the T cell remains viable and trapping the cell directly means that any artefacts due to a cell-bead interface are removed. Our motivation behind this experiment is to use optical tweezers to gain a greater understanding of the interaction forces between T cells and antigen presenting cells. Measuring these interactions has become important due to recent theories which indicate that the strength of this interaction may underlie the activation of the T-cell and subsequent immune response.

  8. Nuclear cardiac imaging for the assessment of myocardial viability.

    Science.gov (United States)

    Slart, R H J A; Bax, J J; van der Wall, E E; van Veldhuisen, D J; Jager, P L; Dierckx, R A

    2005-11-01

    An important aspect of the diagnostic and prognostic work-up of patients with ischaemic cardiomyopathy is the assessment of myocardial viability. Patients with left ventricular dysfunction who have viable myocardium are the patients at highest risk because of the potential for ischaemia but at the same time benefit most from revascularisation. It is important to identify viable myocardium in these patients, and radionuclide myocardial scintigraphy is an excellent tool for this. Single-photon emission computed tomography perfusion scintigraphy (SPECT), whether using 201thallium, 99mTc-sestamibi, or 99mTc- tetrofosmin, in stress and/or rest protocols, has consistently been shown to be an effective modality for identifying myocardial viability and guiding appropriate management. Metabolic and perfusion imaging with positron emission tomography radiotracers frequently adds additional information and is a powerful tool for predicting which patients will have an improved outcome from revascularisation. New techniques in the nuclear cardiology field, such as attenuation corrected SPECT, dual isotope simultaneous acquisition (DISA) SPECT and gated FDG PET are promising and will further improve the detection of myocardial viability. Also the combination of multislice computed tomography scanners with PET opens possibilities of adding coronary calcium scoring and noninvasive coronary angiography to myocardial perfusion imaging and quantification.

  9. Viability of various weed seeds in anaerobic conditions (biogas plant)

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, S.; Hansen, J.

    1983-04-01

    Seeds from different weeds, Urtica urens L. (nettle), Solanum nigrum L. (nightshade), Avena fatua L. (wild oat-grass), Brassica napus L. (rape), Chenopodium album L. (goose-foot), were put into small polyester net bags, which were placed in biogas reactors containing cattle manure. These ''biogas reactors'' were placed at different temperatures . Net bags were taken out after 4.5, 10.5, 21.5, 38 and 53 days, and the seeds were tested for their viability by germination tests and the tetrazolium method. Concerning all seeds it was manifested that the viability decreased very steeply at 35degC. Most of the seeds had a T/sub 50/ at 2-5 days; Chenopodium album L seeds had a T/sub 50/ at 16 days. After 4.5 days it was not possible to find living Avena fatua L seeds. The decrease in viability was less steep at 20degC and even less steep at 2degC.

  10. Dormancy, activation and viability of Rhizopus oligosporus sporangiospores.

    Science.gov (United States)

    Thanh, N V; Nout, M J R

    2004-04-15

    Interruption of dormancy to improve viability of Rhizopus oligosporus sporangiospores is crucial for the application of stored starter cultures for fungal (tempe) production. We aimed to assess the extent of dormancy and factors that could result in activation. Whereas heat treatments were unsuccessful, Malt Extract Broth (MEB) showed to be a good activation medium, with 80% of dormant spores being activated as measured by fluorescence microscopy using a fluorescent marker, compared with 11% with the control. Peptone and yeast extract but not glucose played an important role in activating dormant spores. Metabolically active (fluorescent) and swollen spores, followed by germ tubes were obtained after activation in MEB for 25 min., 2 and 4 h, respectively, at 37 degrees C. Simultaneously, some interesting transitions took place. Dormant spores represent 85-90% of the total spores at harvest and after drying. Their number decreased to 21-32% after activation with MEB with a concomitant increase of metabolically active spores. As a result of storage, some dormancy was lost, yielding an increase of active spores from 11.2% at harvest to 28.8% after 3 months storage. Levels of active spores were well correlated with their viability. By activation of dormant spores, their viability increased; levels of viable and active spores were maximum in 1 month old starter (61.7% and 75.9% of total spores, respectively) but gradually decreased with concomitant increase of the number of dead spores.

  11. Viability And Conidial Production Of Entomopathogenic Fungi Penicillium SP.

    Directory of Open Access Journals (Sweden)

    Nurariaty Agus

    2015-01-01

    Full Text Available Abstract Penicillium sp. order Eurotiales class Eurotiomycetes family Trichocomaceae is one of the entomopathogenic fungi that have the potential to be developed as biological control agent of pests.The study aims to determine the viability and spora production of Entomopathogenic fungi Penicillium sp. Experiments was conducted in Pests Identification and Biological Control laboratory Department of Plant Pest and Disease Faculty of Agriculture Hasanuddin University. The fungus Penicillium sp. cultured in a liquid medium and then added chitin as treatment and others without chitin. The spora viability of fungi was observed on 12th and 24th hours while spora production on 3nd 6th 9th and 12th days after application.The results showed that conidial viability of the fungus Penicillium sp. at 24 hours after application was higher if the medium given chitin than without chitin. The conidial production was higher if given chitin than without chitin. It was highest on 12th day reached 143.4 x 106 conidiaml if media given chitin and on 6th day if without chitin 0.50 x 106 conidiaml.

  12. Cell Viability in Arthroscopic Versus Open Autologous Chondrocyte Implantation.

    Science.gov (United States)

    Biant, Leela C; Simons, Michiel; Gillespie, Trudi; McNicholas, Michael J

    2017-01-01

    Autologous chondrocyte implantation (ACI) is an effective method of repair of articular cartilage defects. It is a 2-stage operation, with the second stage most commonly performed via mini-arthrotomy. Arthroscopic ACI is gaining popularity, as it is less invasive and may accelerate early rehabilitation. However, handling and manipulation of the implant have been shown to cause chondrocyte cell death. To assess the number and viability of cells delivered via an open versus arthroscopic approach in ACI surgery. Controlled laboratory study. Sixteen ACI surgeries were performed on young cadaveric knees by 2 experienced surgeons: 8 via mini-arthrotomy and 8 arthroscopically. Live and dead cells were stained and counted on implants after surgery. The cell number and viability were assessed using confocal laser scanning microscopy. Surgery was timed from knife to skin until the end of cycling the knee 10 times after implantation of the cell-membrane construct. On receipt of cell membranes after transportation from the laboratory, ≥92% of the cells were viable. There were significantly more remaining cells (8.47E+07 arthroscopic vs 1.41E+08 open; P arthroscopic vs 37.34% open; P arthroscopic technique. Open surgery was of a significantly shorter duration (6 vs 32 minutes; P arthroscopic technique. The viability of cells delivered for ACI via an arthroscopic approach was 16 times less than via an open approach. The mini-arthrotomy approach is recommended until long-term clinical comparative data are available.

  13. Vaginal Lactobacilli Reduce Neisseria gonorrhoeae Viability through Multiple Strategies: An in Vitro Study

    Directory of Open Access Journals (Sweden)

    Claudio Foschi

    2017-12-01

    Full Text Available The emergence and spread of antimicrobial resistance in Neisseria gonorrhoeae (GC underline the need of “antibiotic-free” strategies for the control of gonorrhea. The aim of this study was to assess the anti-gonococcal activity of 14 vaginal Lactobacillus strains, belonging to different species (L. crispatus, L. gasseri, L. vaginalis, isolated from healthy pre-menopausal women. In particular, we performed “inhibition” experiments, evaluating the ability of both lactobacilli cells and culture supernatants in reducing GC viability, at two different contact times (7 and 60 min. First, we found that the acidic environment, associated to lactobacilli metabolism, is extremely effective in counteracting GC growth, in a pH- and time-dependent manner. Indeed, a complete abolishment of GC viability by lactobacilli supernatants was observed only for pH values < 4.0, even at short contact times. On the contrary, for higher pH values, no 100%-reduction of GC growth was reached at any contact time. Experiments with organic/inorganic acid solutions confirmed the strict correlation between the pH levels and the anti-gonococcal effect. In this context, the presence of lactate seemed to be crucial for the anti-gonococcal activity, especially for pH values in the range 4.4–5.3, indicating that the presence of H+ ions is necessary but not sufficient to kill gonococci. Moreover, experiments with buffered supernatants led to exclude a direct role in the GC killing by other bioactive molecules produced by lactobacilli. Second, we noticed that lactobacilli cells are able to reduce GC viability and to co-aggregate with gonococci. In this context, we demonstrated that released-surface components with biosurfactant properties, isolated from “highly-aggregating” lactobacilli, could affect GC viability. The antimicrobial potential of biosurfactants isolated from lactobacilli against pathogens has been largely investigated, but this is the first report about a

  14. High-temperature fermentation. How can processes for ethanol production at high temperatures become superior to the traditional process using mesophilic yeast?

    Energy Technology Data Exchange (ETDEWEB)

    Abdel-Banat, Babiker M.A.; Hoshida, Hisashi; Nonklang, Sanom; Akada, Rinji [Yamaguchi Univ. Graduate School of Medicine, Ube (Japan). Dept. of Applied Molecular Bioscience; Ano, Akihiko [Iwata Chemical Co. Ltd. (Japan)

    2010-01-15

    The process of ethanol fermentation has a long history in the production of alcoholic drinks, but much larger scale production of ethanol is now required to enable its use as a substituent of gasoline fuels at 3%, 10%, or 85% (referred to as E3, E10, and E85, respectively). Compared with fossil fuels, the production costs are a major issue for the production of fuel ethanol. There are a number of possible approaches to delivering cost-effective fuel ethanol production from different biomass sources, but we focus in our current report on high-temperature fermentation using a newly isolated thermotolerant strain of the yeast Kluyveromyces marxianus. We demonstrate that a 5 C increase only in the fermentation temperature can greatly affect the fuel ethanol production costs. We contend that this approach may also be applicable to the other microbial fermentations systems and propose that thermotolerant mesophilic microorganisms have considerable potential for the development of future fermentation technologies. (orig.)

  15. Thermotolerant coliform loadings to coastal areas of Santa Catarina (Brazil) evidence the effect of growing urbanisation and insufficient provision of sewerage infrastructure.

    Science.gov (United States)

    Garbossa, Luis H P; Souza, Robson V; Campos, Carlos J A; Vanz, Argeu; Vianna, Luiz F N; Rupp, Guilherme S

    2017-01-01

    Thermotolerant coliform (TC) loadings were quantified for 49 catchments draining into the North and South Bays of Santa Catarina (SC, southeastern Brazil), an area known for its tourism and aquaculture. TC loadings were calculated based on flow measurements taken in 26 rivers. TC concentrations ere quantified based on surface water samples collected at 49 catchment outlets in 2012 and 2013. Median TC loads ranged from 3.7 × 10(3) to 6.8 × 10(8) MPN s(-1). TC loadings in the catchments increased in proportion to increases in resident human population, population density and percentage of urbanised area. Catchments with more than 60% of area covered by wastewater collection and treatment systems had higher TC loads per person than catchments with less than 25%. Based on the study catchments, these results indicate that current sewerage infrastructure is ineffective in reducing contamination of faecal origin to surface waters. These findings have important implications for the management of microbiological health hazards in bathing, recreational and shellfish aquaculture waters in the North and South Bays of Santa Catarina Island.

  16. Study of optimization of wastewater contaminant removal along with extracellular polymeric substances (EPS) production by a thermotolerant Bacillus sp. ISTVK1 isolated from heat shocked sewage sludge.

    Science.gov (United States)

    Gupta, Asmita; Thakur, Indu Shekhar

    2016-08-01

    The present work involved study of wastewater contaminant removal along with EPS production by a thermotolerant bacterium Bacillus sp. ISTVK1, isolated from heat shocked sewage sludge. EPS production in basal and mineral medium containing 50% filter sterilized wastewater and 0.5% sucrose was found to be 0.83±0.12gL(-1) and 0.31±0.10gL(-1) culture, respectively. GC-MS analysis of EPS revealed the presence of β-d-glucose, α-d-galactose and β-d-arabinose. FT-IR spectrum confirmed the presence carbohydrates. Box-Behnken design was used to optimize process parameters for enhanced EPS production along with % COD reduction of wastewater. The optimised conditions when used in a 1.5L bioreactor showed EPS production of 1.67±0.06gL(-1) culture and 93.0±0.21 % COD removal. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Effects of temperature, pH-values and sodium chloride concentrations on the glucose-6-phosphate dehydrogenase activity by thermotolerant Bacillus strains

    Directory of Open Access Journals (Sweden)

    HAZEM AQEL

    2012-01-01

    Full Text Available Thirteen new isolated thermotolerant Bacillus strains and four known Bacillus species were used to evaluate the effect of growth temperature, pH-values and NaCl concentrations on the intracellular glucose-6-phosphate dehydrogenase (G6PDH activity. Results had shown a significant difference in G6PDH production among all species at all used temperatures (p<0.05. The response of individual new isolates and controls for production of G6PDH under growth conditions was variable. The optimal growth conditions did not correspond to the optimal cultivation conditions for maximum G6PDH production. The growth temperature showed the most significant effect on G6PDH activity. The combined effect of temperature and NaCl on the G6PDH activity was strongly pronounced in comparison with the combined effect of temperature and pH or pH and NaCl. Thermal stability at 53ºC and electrophoretic mobility were also investigated. G6PDH from HUTB41 was the most thermostable G6PDH enzyme with T50% of more than 360 minutes. Electrophoretic study demonstrated that G6PDH was composed of two isoenzymes for all strains except B. marinus and B. schlegelii that had three isoenzymes.

  18. Analysis of 16S rRNA and methane monooxygenase gene sequences reveals a novel group of thermotolerant and thermophilic methanotrophs, Methylocaldum gen. nov.

    Science.gov (United States)

    Bodrossy, L; Holmes, E M; Holmes, A J; Kovács, K L; Murrell, J C

    1997-12-01

    Two methanotrophic bacteria with optimum growth temperatures above 40 degrees C were isolated. Thermotolerant strain LK6 was isolated from agricultural soil, and the moderately thermophilic strain OR2 was isolated from the effluent of an underground hot spring. When compared to the described thermophilic methanotrophs Methylococcus capsulatus and Methylococcus thermophilus, these strains are phenotypically similar to Methylococcus thermophilus. However, their 16S rRNA gene sequences are markedly different from the sequence of Methylococcus thermophilus ( approximately 8% divergence) and, together with Methylomonas gracilis, they form a distinct, new genus within the gamma-subgroup of the Proteobacteria related to extant Type I methanotrophs. Further phenotypic characterisation showed that the isolates possess particulate methane monooxygenase (pMMO) but do not contain soluble methane monooxygenase. The nucleotide sequence of a gene encoding pMMO (pmoA) was determined for both isolates and for Methylomonas gracilis. PmoA sequence comparisons confirmed the monophyletic nature of this newly recognised group of thermophilic methanotrophs and their relationship to previously described Type I methanotrophs. We propose that strains OR2 and LK6, together with the misclassified thermophilic strains Methylomonas gracilis VKM-14LT and Methylococcus thermophilus IMV-B3122, comprise a new genus of thermophilic methanotrophs, Methylocaldum gen. nov., containing three new species: Methylocaldum szegediense, Methylocaldum tepidum and Methylocaldum gracile.

  19. Cultivation of two thermotolerant microalgae under tropical conditions: Influences of carbon sources and light duration on biomass and lutein productivity in four seasons.

    Science.gov (United States)

    Chiu, Pai-Ho; Soong, Keryea; Chen, Ching-Nen Nathan

    2016-07-01

    Biomass and lutein productivities of two thermotolerant microalgae were assessed in tropical outdoor conditions in all four seasons. Generally, addition of bicarbonate at 0.2g/L every two days or 2% CO2 did not enhance the productivities compared to the controls, and the productivities in the spring were higher than in the fall. Durations of effective irradiance positively correlated to the productivity of Coelastrella sp. F50 well, but not for Desmodesmus sp. F2. The ineffective light intensity was below 5000 lux (about 106μmol/m(2)s). The productivities produced in the 17cm diameter bottles were 1.5- to 1.9-fold higher than that in the 27cm ones. Lutein content, about 0.5% in biomass on average, did not change significantly grown in different carbon sources or seasons. The annual productivities of the microalgal biomass and lutein in one hectare were estimated to be 33tons and 180kg, respectively, using the non-optimized photobioreactor cultivation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Assessing the Association between Thermotolerant Coliforms in Drinking Water and Diarrhea: An Analysis of Individual–Level Data from Multiple Studies

    Science.gov (United States)

    Hodge, James; Chang, Howard H.; Boisson, Sophie; Collin, Simon M.; Peletz, Rachel; Clasen, Thomas

    2016-01-01

    Background: Fecally contaminated drinking water is believed to be a major contributor to the global burden of diarrheal disease and a leading cause of mortality among young children. However, recent systematic reviews and results from blinded studies of water quality interventions have raised questions about the risk associated with fecally contaminated water, particularly as measured by thermotolerant coliform (TTC) bacteria, a WHO-approved indicator of drinking water quality. Objectives: We investigated the association between TTC in drinking water and diarrhea using data from seven previous studies. Methods: We obtained individual-level data from available field studies that measured TTC levels in household-drinking water and reported prevalence of diarrhea among household members during the days prior to the visit. Results: The combined data set included diarrhea prevalence for 26,518 individuals and 8,000 water samples from 4,017 households, yielding 45,052 observations. The odds of diarrhea increased for each log10 increase in TTC/100 mL by 18% (95% CI: 11, 26%) for children 1,000 TTC/100 mL, respectively compared to coliforms in drinking water and diarrhea: an analysis of individual level data from multiple studies. Environ Health Perspect 124:1560–1567; http://dx.doi.org/10.1289/EHP156 PMID:27164618

  1. The profile of antibiotics resistance and integrons of extended-spectrum beta-lactamase producing thermotolerant coliforms isolated from the Yangtze River basin in Chongqing

    Energy Technology Data Exchange (ETDEWEB)

    Chen Hao [Department of Environmental Hygiene, School of Military Preventive Medicine, Third Military Medical University, 30 Gaotanyan Street, Chongqing 400038 (China); Shu Weiqun, E-mail: west2003@sohu.co [Department of Environmental Hygiene, School of Military Preventive Medicine, Third Military Medical University, 30 Gaotanyan Street, Chongqing 400038 (China); Chang Xiaosong; Chen Jian; Guo Yebin; Tan Yao [Department of Environmental Hygiene, School of Military Preventive Medicine, Third Military Medical University, 30 Gaotanyan Street, Chongqing 400038 (China)

    2010-07-15

    The spreading of extended-spectrum {beta}-lactamases (ESBL)-producing thermotolerant coliforms (TC) in the water environment is a threat to human health but little is known about ESBL-producing TCs in the Yangtze River. We received 319 ESBL-producing stains obtained from the Chongqing basin and we investigated antibiotic susceptibility, bla gene types and the presence of integrons and gene cassettes. 16.8% of TC isolates were ESBL-producing bacteria and bla{sub TEM+CTx-M} was the predominant ESBL type. 65.2% of isolates contained class 1 integrons, but only 3 carried intI 2. Gene cassettes were amplified and sequenced. aadA, drfA, cmlA, sat1, aar3 and two ORF cassettes were found. In conclusion, Yangtze River is heavily polluted by ESBL-producing TC bacteria and the combined bla gene type could enhance antibiotic resistance. Class 1 integrons were widespread in ESBL-producing isolates and play an important role in multi-drug resistance. Characterization of gene cassettes could reveal the dissemination of antibiotic resistance genes. - Yangtze River is heavily polluted by ESBL-producing TC bacteria and Class 1 integrons play an important role in multi-drug resistance.

  2. Assessing the Association between Thermotolerant Coliforms in Drinking Water and Diarrhea: An Analysis of Individual-Level Data from Multiple Studies.

    Science.gov (United States)

    Hodge, James; Chang, Howard H; Boisson, Sophie; Collin, Simon M; Peletz, Rachel; Clasen, Thomas

    2016-10-01

    Fecally contaminated drinking water is believed to be a major contributor to the global burden of diarrheal disease and a leading cause of mortality among young children. However, recent systematic reviews and results from blinded studies of water quality interventions have raised questions about the risk associated with fecally contaminated water, particularly as measured by thermotolerant coliform (TTC) bacteria, a WHO-approved indicator of drinking water quality. We investigated the association between TTC in drinking water and diarrhea using data from seven previous studies. We obtained individual-level data from available field studies that measured TTC levels in household-drinking water and reported prevalence of diarrhea among household members during the days prior to the visit. The combined data set included diarrhea prevalence for 26,518 individuals and 8,000 water samples from 4,017 households, yielding 45,052 observations. The odds of diarrhea increased for each log10 increase in TTC/100 mL by 18% (95% CI: 11, 26%) for children 1,000 TTC/100 mL, respectively compared to coliforms in drinking water and diarrhea: an analysis of individual level data from multiple studies. Environ Health Perspect 124:1560-1567; http://dx.doi.org/10.1289/EHP156.

  3. The profile of antibiotics resistance and integrons of extended-spectrum beta-lactamase producing thermotolerant coliforms isolated from the Yangtze River basin in Chongqing.

    Science.gov (United States)

    Chen, Hao; Shu, Weiqun; Chang, Xiaosong; Chen, Ji-an; Guo, Yebin; Tan, Yao

    2010-07-01

    The spreading of extended-spectrum beta-lactamases (ESBL)-producing thermotolerant coliforms (TC) in the water environment is a threat to human health but little is known about ESBL-producing TCs in the Yangtze River. We received 319 ESBL-producing stains obtained from the Chongqing basin and we investigated antibiotic susceptibility, bla gene types and the presence of integrons and gene cassettes. 16.8% of TC isolates were ESBL-producing bacteria and bla(TEM+CTx-M) was the predominant ESBL type. 65.2% of isolates contained class 1 integrons, but only 3 carried intI 2. Gene cassettes were amplified and sequenced. aadA, drfA, cmlA, sat1, aar3 and two ORF cassettes were found. In conclusion, Yangtze River is heavily polluted by ESBL-producing TC bacteria and the combined bla gene type could enhance antibiotic resistance. Class 1 integrons were widespread in ESBL-producing isolates and play an important role in multi-drug resistance. Characterization of gene cassettes could reveal the dissemination of antibiotic resistance genes. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  4. ClpL is essential for induction of thermotolerance and is potentially part of the HrcA regulon in Lactobacillus gasseri.

    Science.gov (United States)

    Suokko, Aki; Poutanen, Marjo; Savijoki, Kirsi; Kalkkinen, Nisse; Varmanen, Pekka

    2008-03-01

    Stress-inducible proteins are likely to contribute to the survival and activity of probiotic bacteria during industrial processes and in the gastrointestinal tract. The recently published genome sequence of probiotic Lactobacillus gasseri ATCC 33323 suggests the presence of ClpC, ClpE, ClpL, and ClpX from the Clp ATPase family of stress proteins. The heat-shock response of L. gasseri was studied using 2-D DIGE. A total of 20 protein spots showing significant (pgasseri exhibited drastically reduced survival at a lethal temperature and was unable to induce thermotolerance. Genome sequences indicate that the expression of clp genes in several Lactobacillus species is regulated by HrcA, instead of CtsR, the conserved clp gene regulator of low G+C Gram-positive bacteria. Electrophoretic mobility shift assays using L. gasseri HrcA protein and clpL upstream fragments revealed, for the first time, a direct interaction between HrcA and the promoter of a clp gene from a Lactobacillus.

  5. Expression and characterization of thermotolerant lipase with broad pH profiles isolated from an Antarctic Pseudomonas sp strain AMS3

    Directory of Open Access Journals (Sweden)

    Wahhida Latip

    2016-10-01

    Full Text Available A gene encoding a thermotolerant lipase with broad pH was isolated from an Antarctic Pseudomonas strain AMS3. The recombinant lipase AMS3 was purified by single-step purification using affinity chromatography, yielding a purification fold of approximately 1.52 and a recovery of 50%. The molecular weight was approximately ∼60 kDa including the strep and affinity tags. Interestingly, the purified Antarctic AMS3 lipase exhibited broad temperature profile from 10–70 °C and stable over a broad pH range from 5.0 to pH 10.0. Various mono and divalent metal ions increased the activity of the AMS3 lipase, but Ni2+ decreased its activity. The purified lipase exhibited the highest activity in the presence of sunflower oil. In addition, the enzyme activity in 25% v/v solvents at 50 °C particularly to n-hexane, DMSO and methanol could be useful for catalysis reaction in organic solvent and at broad temperature.

  6. Population viability impacts of habitat additions and subtractions: A simulation experiment with endangered kangaroo rats

    Science.gov (United States)

    Species viability is influenced by the quality, quantity and configuration of habitat. For species at risk, a principal challenge is to identify landscape configurations that, if realized, would improve a population’s viability or restoration potential. Critical habitat patche...

  7. Viability of Event Management Business in Batangas City, Philippine: Basis for Business Operation Initiatives

    National Research Council Canada - National Science Library

    Jeninah Christia D. Borbon

    2016-01-01

    The research study on Viability of Event Management Business in Batangas City: Basis for Business Operation Initiatives aimed to assess the viability of this type of business using Thompson’s (2005...

  8. The potential role of polyphenols in the modulation of skin cell viability by Aspalathus linearis and Cyclopia spp. herbal tea extracts in vitro.

    Science.gov (United States)

    Magcwebeba, Tandeka Unathi; Riedel, Sylvia; Swanevelder, Sonja; Swart, Pieter; De Beer, Dalene; Joubert, Elizabeth; Andreas Gelderblom, Wentzel Christoffel

    2016-11-01

    The relationship between polyphenol constituents, antioxidant properties of aqueous and methanol extracts of green tea (Camellia sinensis), the herbal teas, rooibos (Aspalathus linearis) and honeybush (Cyclopia spp.), against skin cell viability was investigated in vitro. The effect of extracts, characterised in terms of polyphenol content and antioxidant properties, on cell viability of premalignant, normal and malignant skin cells was determined. Phenolic composition, particularly high levels of potent antioxidants, of rooibos and green tea methanol extracts was associated with a strong reduction in cell viability specifically targeting premalignant cells. In contrast, the aqueous extracts of Cyclopia spp. were more effective in reducing cell viability. This correlated with a relatively high flavanol/proanthocyanidin content and ABTS radical cation scavenging capacity. The major green tea flavanol (epigallocatechin gallate) and rooibos dihydrochalcone (aspalathin) exhibited differential effects against cell viability, while the major honeybush xanthone (mangiferin) and flavanone (hesperidin) lacked any effect presumably due to a cytoprotective effect. The underlying mechanisms against skin cell viability are likely to involve mitochondrial dysfunction resulting from polyphenol-iron interactions. The polyphenol constituents and antioxidant parameters of herbal tea extracts are useful tools to predict their activity against skin cell survival in vitro and potential chemopreventive effects in vivo. © 2016 Royal Pharmaceutical Society.

  9. Monitoring viability of seeds in gene banks: developing software tools to increase efficiency

    Science.gov (United States)

    Monitoring the decline of seed viability is essential for effective long term seed storage in ex situ collections. Recent FAO Genebank Standards recommend monitoring intervals at one-third the time predicted for viability to fall to 85% of initial viability. This poster outlines the development of ...

  10. Activin Receptor Signaling Regulates Prostatic Epithelial Cell Adhesion and Viability

    Directory of Open Access Journals (Sweden)

    Derek P. Simon

    2009-04-01

    Full Text Available Mutational changes coupled with endocrine, paracrine, and/or autocrine signals regulate cell division during carcinogenesis. The hormone signals remain undefined, although the absolute requirement in vitro for fetal serum indicates the necessity for a fetal serum factor(s in cell proliferation. Using prostatic cancer cell (PCC lines as a model of cancer cell proliferation, we have identified the fetal serum component activin A and its signaling through the activin receptor type II (ActRII, as necessary, although not sufficient, for PCC proliferation. Activin A induced Smad2 phosphorylation and PCC proliferation, but only in the presence of fetal bovine serum (FBS. Conversely, activin A antibodies and inhibin A suppressed FBS-induced PCC proliferation confirming activin A as one of multiple serum components required for PCC proliferation. Basic fibroblast growth factor was subsequently shown to synergize activin A-induced PCC proliferation. Inhibition of ActRII signaling using a blocking antibody or antisense-P decreased mature ActRII expression, Smad2 phosphorylation, and the apparent viability of PCCs and neuroblastoma cells grown in FBS. Suppression of ActRII signaling in PCC and neuroblastoma cells did not induce apoptosis as indicated by the ratio of active/inactive caspase 3 but did correlate with increased cell detachment and ADAM-15 expression, a disintegrin whose expression is strongly correlated with prostatic metastasis. These findings indicate that ActRII signaling is required for PCC and neuroblastoma cell viability, with ActRII mediating cell fate via the regulation of cell adhesion. That ActRII signaling governs both cell viability and cell adhesion has important implications for developing therapeutic strategies to regulate cancer growth and metastasis.

  11. Cell structure and percent viability by a slide centrifuge technique.

    Science.gov (United States)

    Fitzgerald, M G; Hosking, C S

    1982-01-01

    It was found that a slide centrifuge (Cytospin) preparation of a cell suspension allowed a reliable assessment of not only cell structure but also the percentage of non-viable cells. The non-viable cells appeared as "smear" cells and paralleled in number the cells taking up trypan blue. Direct experiment showed the unstained viable cells in a trypan blue cell suspension remained intact in a Cytospin preparation while the cells taking up trypan blue were the "smear" cells. The non-viability of the "smear" cells was confirmed by their inability to survive in culture. Images PMID:7040483

  12. Law and ethics at the border of viability.

    Science.gov (United States)

    Krug, E F

    2006-06-01

    The Supreme Court of Texas in the case of Miller v. HCA announced a rule in 2003 (118 s.w. 3d 758) that a physician attending the delivery of a severely premature infant may provide life-sustaining treatment for that infant under 'emergent circumstances' as a matter of law without first obtaining parental consent. This paper examines issues of law and ethics relevant to decisions about infant resuscitation at the border of viability. It is argued that there is typically no emergency when infants are delivered at 23 weeks gestation, and parents should be asked for informed consent before resuscitation in the delivery room.

  13. Methods for isolation and viability assessment of biological organisms

    Science.gov (United States)

    Letant, Sonia Edith; Baker, Sarah Elyse; Bond, Tiziana; Chang, Allan Shih-Ping

    2015-02-03

    Isolation of biological or chemical organisms can be accomplished using a surface enhanced Raman scattering (SERS) system. The SERS system can be a single or a stacked plurality of photonic crystal membranes with noble-metal lined through pores for flowing analyte potentially containing the biological or chemical organisms. The through pores can be adapted to trap individual biological or chemical organisms and emit SERS spectra, which can then be detected by a detector and further analyzed for viability of the biological or chemical organism.

  14. Viability of human corneal keratocytes during organ culture

    DEFF Research Database (Denmark)

    Møller-Pedersen, T; Møller, H J

    1996-01-01

    The viability of human corneal keratocytes was assessed during four weeks of 'closed system' organ culture at 31 degrees C. After 28 days of culturing, the entire keratocyte population was still alive and viable because all cells incorporated uridine; a parameter for RNA-synthesis. During the fir...... of keratan sulphate proteoglycan suggested that approximately 1% of the total content was lost during the period. In conclusion, our current organ culture technique can maintain a viable keratocyte population for four weeks; a viable stroma can be grafted within this period....

  15. Study of Viability of Phosphate Solubilizing Bacteria in Phosphate granules

    Directory of Open Access Journals (Sweden)

    hajar rajabi

    2017-06-01

    Full Text Available Introduction: sustainable development and the environment are interconnected. Sustainable agriculture is continuous utilization of a farm with respect to various aspects of environmental conditions by using fewer inputs (other than Bio-fertilizers. Phosphorus is one of the essential elements for the plants. Management of soil is possible by using biological fertilizers pillar of sustainable agriculture and providing some of the phosphorus needed by plants via bio-fertilizers. Phosphorus deficiency is extremely effective on the plant growth and productivity. The application of phosphorus fertilizers is expensive and dangerous. In addition, phosphorus in the soilmay become insoluble and will be unavailable to the plants. Studies showed that phosphate solubilizing bacteria in the soil rhizosphere are active and by root exudates solve insoluble phosphates such as tricalcium phosphate, and form absorbable P for plant. Consequently, the use of microbial fertilizers could reduce excessive use of chemical fertilizers and lead to decrease their harmful effects and protect the environment and conservation of available resources. The biological phosphate fertilizer industry uses sugar beet molasses as a binder and drying granules at high temperatures. Therefore, it is important to evaluate the durability of the bacteria in molasses at high temperature. Materials and Methods: This study was designed as completely randomized design in a factorial arrangement.10 isolates were selected and the ratios of 50%, 25%, 15% and 10% of the apatite, organic matter, sulfur and soluble granule (ratio 1: 1 and 2: 1 bacteria and molasses, respectively, for each isolate was prepared. The final product was dried at 28 and 40 °C and remained for 4 months and population counted at first day and 10, 20, 30, 60, 90 and 120 days after the preparing. The population was counted by the serial dilution technique and cultured at Sperber media. Results and Discussion:Comparing the

  16. Simulated Microgravity and 3D Culture Enhance Induction, Viability, Proliferation and Differentiation of Cardiac Progenitors from Human Pluripotent Stem Cells

    Science.gov (United States)

    Jha, Rajneesh; Wu, Qingling; Singh, Monalisa; Preininger, Marcela K.; Han, Pengcheng; Ding, Gouliang; Cho, Hee Cheol; Jo, Hanjoong; Maher, Kevin O.; Wagner, Mary B.; Xu, Chunhui

    2016-01-01

    Efficient generation of cardiomyocytes from human pluripotent stem cells is critical for their regenerative applications. Microgravity and 3D culture can profoundly modulate cell proliferation and survival. Here, we engineered microscale progenitor cardiac spheres from human pluripotent stem cells and exposed the spheres to simulated microgravity using a random positioning machine for 3 days during their differentiation to cardiomyocytes. This process resulted in the production of highly enriched cardiomyocytes (99% purity) with high viability (90%) and expected functional properties, with a 1.5 to 4-fold higher yield of cardiomyocytes from each undifferentiated stem cell as compared with 3D-standard gravity culture. Increased induction, proliferation and viability of cardiac progenitors as well as up-regulation of genes associated with proliferation and survival at the early stage of differentiation were observed in the 3D culture under simulated microgravity. Therefore, a combination of 3D culture and simulated microgravity can be used to efficiently generate highly enriched cardiomyocytes. PMID:27492371

  17. Viability of Ascaris suum eggs in stored raw and separated liquid slurry.

    Science.gov (United States)

    Katakam, Kiran Kumar; Roepstorff, Allan; Popovic, Olga; Kyvsgaard, Niels C; Thamsborg, Stig Milan; Dalsgaard, Anders

    2013-03-01

    Separation of pig slurry into solid and liquid fractions is gaining importance as a way to manage increasing volumes of slurry. In contrast to solid manure and slurry, little is known about pathogen survival in separated liquid slurry. The viability of Ascaris suum eggs, a conservative indicator of fecal pollution, and its association with ammonia was investigated in separated liquid slurry in comparison with raw slurry. For this purpose nylon bags with 6000 eggs each were placed in 1 litre bottles containing one of the two fractions for 308 days at 5 °C or 25 °C. Initial analysis of helminth eggs in the separated liquid slurry revealed 47 Ascaris eggs per gramme. At 25 °C, egg viability declined to zero with a similar trend in both raw slurry and the separated liquid slurry by day 308, a time when at 5 °C 88% and 42% of the eggs were still viable in separated liquid slurry and raw slurry, respectively. The poorer survival at 25 °C was correlated with high ammonia contents in the range of 7.9-22.4 mM in raw slurry and 7.3-23.2 mM in liquid slurry compared to 3.2-9.5 mM in raw slurry and 2.6-9.5 mM in liquid slurry stored at 5 °C. The study demonstrates that at 5 °C, A. suum eggs have a higher viability in separated liquid slurry as compared to raw slurry. The hygiene aspect of this needs to be further investigated when separated liquid slurry is used to fertilize pastures or crops.

  18. A novel concentration and viability detection method for Brettanomyces using the Cellometer image cytometry.

    Science.gov (United States)

    Martyniak, Brian; Bolton, Jason; Kuksin, Dmitry; Shahin, Suzanne M; Chan, Leo Li-Ying

    2017-01-01

    Brettanomyces spp. can present unique cell morphologies comprised of excessive pseudohyphae and budding, leading to difficulties in enumerating cells. The current cell counting methods include manual counting of methylene blue-stained yeasts or measuring optical densities using a spectrophotometer. However, manual counting can be time-consuming and has high operator-dependent variations due to subjectivity. Optical density measurement can also introduce uncertainties where instead of individual cells counted, an average of a cell population is measured. In contrast, by utilizing the fluorescence capability of an image cytometer to detect acridine orange and propidium iodide viability dyes, individual cell nuclei can be counted directly in the pseudohyphae chains, which can improve the accuracy and efficiency of cell counting, as well as eliminating the subjectivity from manual counting. In this work, two experiments were performed to demonstrate the capability of Cellometer image cytometer to monitor Brettanomyces concentrations, viabilities, and budding/pseudohyphae percentages. First, a yeast propagation experiment was conducted to optimize software counting parameters for monitoring the growth of Brettanomyces clausenii, Brettanomyces bruxellensis, and Brettanomyces lambicus, which showed increasing cell concentrations, and varying pseudohyphae percentages. The pseudohyphae formed during propagation were counted either as multiple nuclei or a single multi-nuclei organism, where the results of counting the yeast as a single multi-nuclei organism were directly compared to manual counting. Second, a yeast fermentation experiment was conducted to demonstrate that the proposed image cytometric analysis method can monitor the growth pattern of B. lambicus and B. clausenii during beer fermentation. The results from both experiments displayed different growth patterns, viability, and budding/pseudohyphae percentages for each Brettanomyces species. The proposed

  19. Viability of calcifying bacterial formulations in fly ash for applications in building materials.

    Science.gov (United States)

    Dhami, Navdeep Kaur; Mukherjee, Abhijit; Reddy, M Sudhakara

    2013-12-01

    Evidence of bacterial involvement in precipitation of calcium carbonates has brought a revolution in the field of applied microbiology, geotechnical sciences, environmental and civil engineering with its marked success in restoration of various building materials. For applications of these calcite binder-producing bacterial cultures, different expensive carrier materials have been used but their high costs have come in the way of their successful commercialization. In the present study, we have explored the potential of cheap industrial by-product fly ash as a carrier material for bacterial cells and investigated the viability of calcifying bacterial isolates: Bacillus megaterium, Bacillus cereus, and Lysinibacillus fusiformis in fly ash carrier at varying temperatures and moisture conditions along with biomineralization efficacy of these formulations. We used laser scanning confocal microscopy to analyze the viability of bacteria by florescent dye 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) along with the plate count method. Results revealed that fly ash successfully served as an effective carrier material and bacterial formulations stored at 4 °C provided longer shelf life than those stored at higher temperatures. Up to 10(6) cfu/g was found to sustain in all formulations at 4 °C compared to 10(4)-10(5) cfu/g in case of higher temperatures up to 1 year. For 4 °C, higher moistures (50 %) were found to provide better survivability while for higher temperatures, lower moistures (30 %) favored higher viability. The biomineralization capability of fresh and formulated bacterial cells was compared on the basis of precipitation of carbonates and it was found that carbonate precipitation efficacy of formulated bacterial cells was comparable to fresh bacterial cells.

  20. Viability of Booby Offspring is Maximized by Having One Young Parent and One Old Parent.

    Directory of Open Access Journals (Sweden)

    Hugh Drummond

    Full Text Available It is widely expected that the quality of offspring will vary with the age of their parents and that this variation should influence animals' choice of mates. However, theoretical predictions for age effects are contradictory and, to our knowledge, we do not know for any wild animal how the quality of offspring is affected by both parents' ages across their lifespans, or whether mothers' and fathers' ages interact. We tackled this question using long-term data on a highly philopatric, insular population of the blue-footed booby (Sula nebouxii. In this species extra-pair paternity is most common in pairs of two young parents or two old parents, implying that these age combinations might prejudice offspring quality. Analysis of the viability of 3,361 offspring of parents up to 21 years old revealed that fledglings with two young parents or two old parents were least likely to become breeders, whereas fledglings with one young parent and one old parent were most likely to do so. For young parents of either sex, offspring viability increased with age of the other parent; for very old parents, it decreased. These effects could be mediated by parents flexibly modifying their investment in offspring in response to their own and their partners´ ages, but evidence for this was lacking. In 5,343 breeding attempts, although mothers' and fathers' ages independently affected four heavily care-dependent breeding traits at the clutch and nestling stages, their interaction did not affect any trait. The effects of parental age combinations on viability could also be mediated by genes: fledglings with one young parent and one old parent could benefit from greater heterozygosity or better genes.

  1. Detection of myocardial viability by low-dose dobutamine Cine MR imaging.

    Science.gov (United States)

    Sandstede, J J; Bertsch, G; Beer, M; Kenn, W; Werner, E; Pabst, T; Lipke, C; Kretschmer, S; Neubauer, S; Hahn, D

    1999-12-01

    The purpose of this work was to test the diagnostic value of dobutamine stress magnetic resonance imaging (MRI) for predicting recovery of regional myocardial contractility after revascularization. Cardiac wall motion abnormalities are due to either non-viable and/or scarred, or viable, but hibernating, myocardial tissue. Dobutamine stress leads to increased systolic wall thickening only in viable myocardium. Twenty-five patients with akinetic or dyskinetic myocardial regions were examined with a Cine FLASH-2D sequence at rest and during dobutamine stress (10 microg/kg/min). Patients were re-examined at rest 3, and in case of persisting wall motion defects, 6 months after revascularization. Criterion of viability was increasing end-systolic wall thickening during stress and/or at follow-up. Akinetic regions related either to the LAD (n = 19) or to the RCA (n = 6) were judged viable if > or = 50% of the affected segments improved. MR studies were completed in all subjects without arrhythmia or need for early terminations due to symptoms. Sensitivity, specificity, and positive predictive value for the prediction of myocardial viability were 61%, 90%, and 87% for the segment-related analysis, and 76%, 100%, and 100% for the patient-related analysis based on coronary artery distribution, respectively. Dobutamine stress MRI allows to predict global functional recovery of akinetic myocardial regions after revascularization with a high positive predictive value and high specificity.

  2. Effect of chitosan and SO2 on viability of Acetobacter strains in wine.

    Science.gov (United States)

    Valera, Maria José; Sainz, Florencia; Mas, Albert; Torija, María Jesús

    2017-04-04

    Wine spoilage is an important concern for winemakers to preserve the quality of their final product and avoid contamination throughout the production process. The use of sulphur dioxide (SO2) is highly recommended to prevent wine spoilage due to its antimicrobial activity. However, SO2 has a limited effect on the viability of acetic acid bacteria (AAB). Currently, the use of SO2 alternatives is favoured in order to reduce the use of chemicals and improve stabilization in winemaking. Chitosan is a biopolymer that is approved by the European authorities and the International Organization of Vine and Wine to be used as a fining agent and antimicrobial in wines. However, its effectiveness in AAB prevention has not been studied. Two strains of Acetobacter, adapted to high ethanol environments, were analysed in this study. Both chitosan and SO2 effects were compared in artificially contaminated wines. Both molecules reduced the metabolic activity of both AAB strains. Although AAB populations were detected by culture independent techniques, their numbers were reduced with time, and their viability decreased following the application of both products, especially with chitosan. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Resuscitation at the limits of viability--an Irish perspective.

    LENUS (Irish Health Repository)

    Khan, R A

    2012-02-01

    BACKGROUND: Advances in neonatal care continue to lower the limit of viability. Decision making in this grey zone remains a challenging process. OBJECTIVE: To explore the opinions of healthcare providers on resuscitation and outcome in the less than 28-week preterm newborn. DESIGN\\/METHODS: An anonymous postal questionnaire was sent to health care providers working in maternity units in the Republic of Ireland. Questions related to neonatal management of the extreme preterm infant, and estimated survival and long-term outcome. RESULTS: The response rate was 55% (74% obstetricians and 70% neonatologists). Less than 1% would advocate resuscitation at 22 weeks, 10% of health care providers advocate resuscitation at 23 weeks gestation, 80% of all health care providers would resuscitate at 24 weeks gestation. 20% of all health care providers would advocate cessation of resuscitation efforts on 22-25 weeks gestation at 5 min of age. 65% of Neonatologists and 54% trainees in Paediatrics would cease resuscitation at 10 min of age. Obstetricians were more pessimistic about survival and long term outcome in newborns delivered between 23 and 27 weeks when compared with neonatologists. This difference was also observed in trainees in paediatrics and obstetrics. CONCLUSION: Neonatologists, trainees in paediatrics and neonatal nurses are generally more optimistic about outcome than their counterparts in obstetrical care and this is reflected in a greater willingness to provide resuscitation efforts at the limits of viability.

  4. Viability of Hanseniaspora uvarum yeast preserved by lyophilization and cryopreservation.

    Science.gov (United States)

    de Arruda Moura Pietrowski, Giovana; Grochoski, Mayara; Sartori, Gabriela Felkl; Gomes, Tatiane Aparecida; Wosiacki, Gilvan; Nogueira, Alessandro

    2015-08-01

    Hanseniaspora yeasts are known to produce volatile compounds that give fruity aromas in wine and fermented fruit. This study aimed to verify the feasibility of the Hanseniaspora uvarum strain that had been isolated and identified during a previous study and preserved by lyophilization and freezing at -80 °C (cryopreservation). This strain was assessed in relation to its macroscopic and microscopic morphology and for its ability to ferment apple must. After having been subjected to lyophilization and cryopreservation, viability was assessed in relation to these characteristics during 12 months of storage. The strain showed stable colonial features and its microscopic appearance was unchanged during all recoveries. The plate count results showed consistency in both processes. Regarding the fermentative capacity, the kinetic results showed 100% viability for the strain subjected to lyophilization, as well as for those preserved at -80 °C. These results demonstrate that the preservation methods used are compatible with the maintenance of the relevant characteristics of the strain for the period of evaluation of this study (12 months). Copyright © 2015 John Wiley & Sons, Ltd.

  5. Improvement on The Ellis and Roberts Viability Model

    Directory of Open Access Journals (Sweden)

    Guoyan Zhou

    2016-05-01

    Full Text Available With data sets of germination percent and storage time of seed lot of wheat and sorghum stored at three different storage temperature(t, °C with three different water content (m, % of seeds, together with data set of buckwheat and lettuce reported in literatures, the possibility that seed survival curve were transformed into line by survival proportion and the relationship that logarithm of average viability period (logp50 and standard deviation of seed death distribution in time (δwith t, m and interaction between t and m were analysed. Result indicated that survival proportion transformed seed survival curve to line were much easier than the probability adopted by Ellis and Roberts, and the most important factor affecting logp50 and δ of seed lot was interaction between t and m. Thus, Ellis and Roberts viability model were suggested to be improved as Ki=Vi-p/10K-CWT (t×m to predict longevity of seed lot with initial germination percent unknown, a new model of Gi/G0=A-P/10K-CWT(t×m was constructed to predict longevity of seed lot with initial germination percent already known.

  6. Economic viability of cerrado vegetation management under conditions of risk

    Directory of Open Access Journals (Sweden)

    Simão Corrêa da Silva

    2011-06-01

    Full Text Available Cerrado vegetation is Brazil’s second largest biome, comprising about 388 municipalities in Minas Gerais state alone and serving as an important source of natural resources. A large share of the wood charcoal produced in Minas Gerais is sourced from Cerrado vegetation. The objective of this work is to assess the economic viability of Cerrado vegetation management for wood charcoal production, under conditions of risk. The study site is a fragment of Cerrado subjected to five levels of intervention as to basal area removal. For risk analysis, the Monte Carlo method was applied, using charcoal price, interest rate and land value as input variables, and using Net Present Value as output variable over an infinite planning horizon. It was concluded that introducing risk in the economic analysis of the various Cerrado management regimes helped provide additional information to that obtained by deterministic analysis, improving understanding and ensuring safety in decision-making about the economic viability of such regimes. For all treatments, the probability of VPL being negative increases with increasing cutting cycle lengths. For all treatments, the optimal cutting cycle is ten years. Treatments where a larger volume of wood was removed proved less prone to risks of economic inviability since they secure more revenue than treatments where less wood was removed.

  7. Cell viability and angiogenic potential of a bioartificial adipose substitute.

    Science.gov (United States)

    Panneerselvan, Anitha; Nguyen, Luong T H; Su, Yan; Teo, Wee Eong; Liao, Susan; Ramakrishna, Seeram; Chan, Ching Wan

    2015-06-01

    An implantable scaffold pre-seeded with cells needs to remain viable and encourage rapid angiogenesis in order to replace injured tissues, especially for tissue defect repairs. We created a bioartificial adipose graft composed of an electrospun 3D nanofibrous scaffold and fat tissue excised from New Zealand white rabbits. Cell viability and angiogenesis potential of the bioartificial substitute were examined during four weeks of culture in Dulbecco's Modified Eagle Medium by immunohistochemical staining with LIVE/DEAD® cell kit and PECAM-1 antibody, respectively. In addition, a Matrigel® assay was performed to examine the possibility of blood vessels sprouting from the bioartificial graft. Our results showed that cells within the graft were viable and vascular tubes were present at week 4, while cells in a fat tissue block were dead in vitro. In addition, capillaries were observed sprouting from the graft into the Matrigel, demonstrating its angiogenic potential. We expect that improved cell viability and angiogenesis in the bioartificial substitute, compared to intact autologous graft, could potentially contribute to its survival following implantation. Copyright © 2012 John Wiley & Sons, Ltd.

  8. The postmitotic Saccharomyces cerevisiae after spaceflight showed higher viability

    Science.gov (United States)

    Yi, Zong-Chun; Li, Xiao-Fei; Wang, Yan; Wang, Jie; Sun, Yan; Zhuang, Feng-Yuan

    2011-06-01

    The budding yeast Saccharomyces cerevisiae has been proposed as an ideal model organism for clarifying the biological effects caused by spaceflight conditions. The postmitotic S. cerevisiae cells onboard Practice eight recoverable satellite were subjected to spaceflight for 15 days. After recovery, the viability, the glycogen content, the activities of carbohydrate metabolism enzymes, the DNA content and the lipid peroxidation level in yeast cells were analyzed. The viability of the postmitotic yeast cells after spaceflight showed a three-fold increase as compared with that of the ground control cells. Compared to the ground control cells, the lipid peroxidation level in the spaceflight yeast cells markedly decreased. The spaceflight yeast cells also showed an increase in G2/M cell population and a decrease in Sub-G1 cell population. The glycogen content and the activities of hexokinase and succinate dehydrogenase significantly decreased in the yeast cells after spaceflight. In contrast, the activity of malate dehydrogenase showed an obvious increase after spaceflight. These results suggested that microgravity or spaceflight could promote the survival of postmitotic S. cerevisiae cells through regulating carbohydrate metabolism, ROS level and cell cycle progression.

  9. Production and economic viability of feedlot beef cattle categories

    Directory of Open Access Journals (Sweden)

    Petrônio Batista dos Santos

    2017-04-01

    Full Text Available This study was realized to evaluate the production and economic viability of finished beef cattle in feedlot in the Cerrado biome of Piauí State, Brazil. One hundred and fifty cattle -50 bulls (B with an body weight of 283.0 ± 20.82 kg, at 30 months of age; and 100 cull cows (CC with an body weight of 296.1 ± 17.80 kg, at 100 months of age- were used in a completely randomized design. The animal performance indicators for the evaluation of economic viability were initial age (months, slaughter age (months, was initial body weight (kg, final body weight (kg, daily feed intake (kg animal-1; kg kg-1 BW, diet cost (R$ kg-1 of diet, cost of kg produced (R$ kg-1 produced, and average price of the kg of meat for finishing (R$. The CC consumed more sorghum silage and concentrate than B (p < 0.05.There was a difference between the categories (p < 0.05 for gross revenue, balance, opportunity cost, and net revenue per animal, with higher values found for the steer category. The animal category did not affect the profitability of the system, although the bulls provided lower revenues than cull cows.

  10. Noninvasive diagnosis of seed viability using infrared thermography.

    Science.gov (United States)

    Kranner, Ilse; Kastberger, Gerald; Hartbauer, Manfred; Pritchard, Hugh W

    2010-02-23

    Recent advances in the noninvasive analyses of plant metabolism include stress imaging techniques, mainly developed for vegetative tissues. We explored if infrared thermography can be used to predict whether a quiescent seed will germinate or die upon water uptake. Thermal profiles of viable, aged, and dead Pisum sativum seeds were recorded, and image analysis of 22,000 images per individual seed showed that infrared thermography can detect imbibition- and germination-associated biophysical and biochemical changes. These "thermal fingerprints" vary with viability in this species and in Triticum aestivum and Brassica napus seeds. Thermogenesis of the small individual B. napus seeds was at the limit of the technology. We developed a computer model of "virtual pea seeds," that uses Monte Carlo simulation, based on the heat production of major seed storage compounds to unravel physico-chemical processes of thermogenesis. The simulation suggests that the cooling that dominates the early thermal profiles results from the dissolution of low molecular-weight carbohydrates. Moreover, the kinetics of the production of such "cooling" compounds over the following 100 h is dependent on seed viability. We also developed a deterministic tool that predicts in the first 3 hours of water uptake, when seeds can be redried and stored again, whether or not a pea seed will germinate. We believe that the early separation of individual, ungerminated seeds (live, aged, or dead) before destructive germination assessment creates unique opportunities for integrative studies on cell death, differentiation, and development.

  11. Effects of drinking desalinated seawater on cell viability and proliferation.

    Science.gov (United States)

    Macarrão, Camila Longhi; Bachi, André Luis Lacerda; Mariano, Mario; Abel, Lucia Jamli

    2017-06-01

    Desalination of seawater is becoming an important means to address the increasing scarcity of freshwater resources in the world. Seawater has been used as drinking water in the health, food, and medical fields and various beneficial effects have been suggested, although not confirmed. Given the presence of 63 minerals and trace elements in drinking desalinated seawater (63 DSW), we evaluated their effects on the behavior of tumorigenic and nontumorigenic cells through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and annexin-V-fluorescein isothiocyanate/propidium iodide staining. Our results showed that cell viability and proliferation in the presence of 63 DSW were significantly greater than in mineral water and in the presence of fetal bovine serum in a dose-dependent manner. Furthermore, 63 DSW showed no toxic effect on murine embryonic fibroblast (NIH-3T3) and murine melanoma (B16-F10) cells. In another assay, we also showed that pre-treatment of non-adherent THP-1 cells with 63 DSW reduces apoptosis incidence, suggesting a protective effect against cell death. We conclude that cell viability and proliferation were improved by the mineral components of 63 DSW and this effect can guide further studies on health effects associated with DSW consumption.

  12. Noninvasive diagnosis of seed viability using infrared thermography

    Science.gov (United States)

    Kranner, Ilse; Kastberger, Gerald; Hartbauer, Manfred; Pritchard, Hugh W.

    2010-01-01

    Recent advances in the noninvasive analyses of plant metabolism include stress imaging techniques, mainly developed for vegetative tissues. We explored if infrared thermography can be used to predict whether a quiescent seed will germinate or die upon water uptake. Thermal profiles of viable, aged, and dead Pisum sativum seeds were recorded, and image analysis of 22,000 images per individual seed showed that infrared thermography can detect imbibition- and germination-associated biophysical and biochemical changes. These “thermal fingerprints” vary with viability in this species and in Triticum aestivum and Brassica napus seeds. Thermogenesis of the small individual B. napus seeds was at the limit of the technology. We developed a computer model of “virtual pea seeds,” that uses Monte Carlo simulation, based on the heat production of major seed storage compounds to unravel physico-chemical processes of thermogenesis. The simulation suggests that the cooling that dominates the early thermal profiles results from the dissolution of low molecular-weight carbohydrates. Moreover, the kinetics of the production of such “cooling” compounds over the following 100 h is dependent on seed viability. We also developed a deterministic tool that predicts in the first 3 hours of water uptake, when seeds can be redried and stored again, whether or not a pea seed will germinate. We believe that the early separation of individual, ungerminated seeds (live, aged, or dead) before destructive germination assessment creates unique opportunities for integrative studies on cell death, differentiation, and development. PMID:20133712

  13. Effect of Antarctic solar radiation on sewage bacteria viability.

    Science.gov (United States)

    Hughes, Kevin A

    2005-06-01

    The majority of coastal Antarctic research stations discard untreated sewage waste into the near-shore marine environment. However, Antarctic solar conditions are unique, with ozone depletion increasing the proportion of potentially damaging ultraviolet-B (UV-B) radiation reaching the marine environment. This study assessed the influence of Antarctic solar radiation on the viability of Escherichia coli and sewage microorganisms at Rothera Research Station, Adelaide Island, Antarctic Peninsula. Cell viability decreased with increased exposure time and with exposure to shorter wavelengths of solar radiation. Cell survival also declined with decreasing cloud cover, solar zenith angle and ozone column depth. However, particulates in sewage increased the persistence of viable bacteria. Ultraviolet radiation doses over Rothera Point were highest during the austral summer. During this time, solar radiation may act to partially reduce the number of viable sewage-derived microorganisms in the surface seawater around Antarctic outfalls. Nevertheless, this effect is not reliable and every effort should be made to fully treat sewage before release into the Antarctic marine environment.

  14. Sporosalibacterium tautonense sp. nov., a thermotolerant, halophilic, hydrolytic bacterium isolated from a gold mine, and emended description of the genus Sporosalibacterium.

    Science.gov (United States)

    Podosokorskaya, Olga A; Merkel, Alexander Y; Heerden, Esta van; Cason, Errol D; Kopitsyn, Dmitry S; Vasilieva, Maria; Bonch-Osmolovskaya, Elizaveta A; Kublanov, Ilya V

    2017-05-01

    A novel strictly anaerobic, thermotolerant, moderately halophilic, organotrophic bacterium, strain MRo-4T, was isolated from a sample of a microbial mat, developed under the flow of subsurface water in TauTona gold mine, South Africa. Cells of the novel isolate stained Gram-positive and were motile, spore-forming rods, 0.2-0.3 µm in width and 5-20 µm in length. Strain MRo-4T grew at 25-50 °C, at pH 7.0-8.8 and at an NaCl concentration of 5-100 g l-1. The isolate was able to ferment yeast extract, peptone and mono-, oligo- and polysaccharides, including cellulose and chitin. Elemental sulfur, thiosulfate, sulfate, sulfite, nitrate, nitrite, fumarate and arsenate were not reduced. The major fatty acids were iso-C15 : 0, iso-C15 : 0 dimethyl acetyl and anteiso-C15 : 0. The G+C content of the DNA was 32.9 mol%. Phylogenetic analysis of 16S rRNA gene sequences of strain MRo-4T and its nearest relatives showed its affiliation to the genus Sporosalibacterium. Sporosalibacteriumfaouarense SOL3f37T, the only valid published representative of the genus, appeared to be its closest relative (96.8 % 16S rRNA gene sequence similarity). However, strains MRo-4T and S. faouarense SOL3f37T differed in temperature, pH and salinity ranges for growth, requirement for yeast extract and substrate profiles. Based on the phylogenetic analysis and physiological properties of the novel isolate, we propose a novel species, Sporosalibacterium tautonense sp. nov. The type strain is MRo-4T (=DSM 28179T=VKM B-2948T).

  15. Plantation forestry under global warming: hybrid poplars with improved thermotolerance provide new insights on the in vivo function of small heat shock protein chaperones.

    Science.gov (United States)

    Merino, Irene; Contreras, Angela; Jing, Zhong-Ping; Gallardo, Fernando; Cánovas, Francisco M; Gómez, Luis

    2014-02-01

    Climate-driven heat stress is a key factor affecting forest plantation yields. While its effects are expected to worsen during this century, breeding more tolerant genotypes has proven elusive. We report here a substantial and durable increase in the thermotolerance of hybrid poplar (Populus tremula×Populus alba) through overexpression of a major small heat shock protein (sHSP) with convenient features. Experimental evidence was obtained linking protective effects in the transgenic events with the unique chaperone activity of sHSPs. In addition, significant positive correlations were observed between phenotype strength and heterologous sHSP accumulation. The remarkable baseline levels of transgene product (up to 1.8% of total leaf protein) have not been reported in analogous studies with herbaceous species. As judged by protein analyses, such an accumulation is not matched either by endogenous sHSPs in both heat-stressed poplar plants and field-grown adult trees. Quantitative real time-polymerase chain reaction analyses supported these observations and allowed us to identify the poplar members most responsive to heat stress. Interestingly, sHSP overaccumulation was not associated with pleiotropic effects that might decrease yields. The poplar lines developed here also outperformed controls under in vitro and ex vitro culture conditions (callus biomass, shoot production, and ex vitro survival), even in the absence of thermal stress. These results reinforce the feasibility of improving valuable genotypes for plantation forestry, a field where in vitro recalcitrance, long breeding cycles, and other practical factors constrain conventional genetic approaches. They also provide new insights into the biological functions of the least understood family of heat shock protein chaperones.

  16. Plantation Forestry under Global Warming: Hybrid Poplars with Improved Thermotolerance Provide New Insights on the in Vivo Function of Small Heat Shock Protein Chaperones1[C][W

    Science.gov (United States)

    Merino, Irene; Contreras, Angela; Jing, Zhong-Ping; Gallardo, Fernando; Cánovas, Francisco M.; Gómez, Luis

    2014-01-01

    Climate-driven heat stress is a key factor affecting forest plantation yields. While its effects are expected to worsen during this century, breeding more tolerant genotypes has proven elusive. We report here a substantial and durable increase in the thermotolerance of hybrid poplar (Populus tremula × Populus alba) through overexpression of a major small heat shock protein (sHSP) with convenient features. Experimental evidence was obtained linking protective effects in the transgenic events with the unique chaperone activity of sHSPs. In addition, significant positive correlations were observed between phenotype strength and heterologous sHSP accumulation. The remarkable baseline levels of transgene product (up to 1.8% of total leaf protein) have not been reported in analogous studies with herbaceous species. As judged by protein analyses, such an accumulation is not matched either by endogenous sHSPs in both heat-stressed poplar plants and field-grown adult trees. Quantitative real time-polymerase chain reaction analyses supported these observations and allowed us to identify the poplar members most responsive to heat stress. Interestingly, sHSP overaccumulation was not associated with pleiotropic effects that might decrease yields. The poplar lines developed here also outperformed controls under in vitro and ex vitro culture conditions (callus biomass, shoot production, and ex vitro survival), even in the absence of thermal stress. These results reinforce the feasibility of improving valuable genotypes for plantation forestry, a field where in vitro recalcitrance, long breeding cycles, and other practical factors constrain conventional genetic approaches. They also provide new insights into the biological functions of the least understood family of heat shock protein chaperones. PMID:24306533

  17. BPM-CUL3 E3 ligase modulates thermotolerance by facilitating negative regulatory domain-mediated degradation of DREB2A in Arabidopsis.

    Science.gov (United States)

    Morimoto, Kyoko; Ohama, Naohiko; Kidokoro, Satoshi; Mizoi, Junya; Takahashi, Fuminori; Todaka, Daisuke; Mogami, Junro; Sato, Hikaru; Qin, Feng; Kim, June-Sik; Fukao, Yoichiro; Fujiwara, Masayuki; Shinozaki, Kazuo; Yamaguchi-Shinozaki, Kazuko

    2017-10-03

    DEHYDRATION-RESPONSIVE ELEMENT BINDING PROTEIN 2A (DREB2A) acts as a key transcription factor in both drought and heat stress tolerance in Arabidopsis and induces the expression of many drought- and heat stress-inducible genes. Although DREB2A expression itself is induced by stress, the posttranslational regulation of DREB2A, including protein stabilization, is required for its transcriptional activity. The deletion of a 30-aa central region of DREB2A known as the negative regulatory domain (NRD) transforms DREB2A into a stable and constitutively active form referred to as DREB2A CA. However, the molecular basis of this stabilization and activation has remained unknown for a decade. Here we identified BTB/POZ AND MATH DOMAIN proteins (BPMs), substrate adaptors of the Cullin3 (CUL3)-based E3 ligase, as DREB2A-interacting proteins. We observed that DREB2A and BPMs interact in the nuclei, and that the NRD of DREB2A is sufficient for its interaction with BPMs. BPM-knockdown plants exhibited increased DREB2A accumulation and induction of DREB2A target genes under heat and drought stress conditions. Genetic analysis indicated that the depletion of BPM expression conferred enhanced thermotolerance via DREB2A stabilization. Thus, the BPM-CUL3 E3 ligase is likely the long-sought factor responsible for NRD-dependent DREB2A degradation. Through the negative regulation of DREB2A stability, BPMs modulate the heat stress response and prevent an adverse effect of excess DREB2A on plant growth. Furthermore, we found the BPM recognition motif in various transcription factors, implying a general contribution of BPM-mediated proteolysis to divergent cellular responses via an accelerated turnover of transcription factors.

  18. Ethanol production from alkali-treated rice straw via simultaneous saccharification and fermentation using newly isolated thermotolerant Pichia kudriavzevii HOP-1.

    Science.gov (United States)

    Oberoi, Harinder Singh; Babbar, Neha; Sandhu, Simranjeet Kaur; Dhaliwal, Sandeep Singh; Kaur, Ujjal; Chadha, B S; Bhargav, Vinod Kumar

    2012-04-01

    In this study, simultaneous saccharification and fermentation (SSF) was employed to produce ethanol from 1% sodium hydroxide-treated rice straw in a thermostatically controlled glass reactor using 20 FPU gds⁻¹ cellulase, 50 IU gds⁻¹ β-glucosidase, 15 IU gds⁻¹ pectinase and a newly isolated thermotolerant Pichia kudriavzevii HOP-1 strain. Scanning electron micrograph images showed that the size of the P. kudriavzevii cells ranged from 2.48 to 6.93 μm in diameter while the shape of the cells varied from oval, ellipsoidal to elongate. Pichia kudriavzevii cells showed extensive pseudohyphae formation after 5 days of growth and could assimilate sugars like glucose, sucrose, galactose, fructose, and mannose but the cells could not assimilate xylose, arabinose, cellobiose, raffinose, or trehalose. In addition, the yeast cells could tolerate up to 40% glucose and 5% NaCl concentrations but their growth was inhibited at 1% acetic acid and 0.01% cyclohexamide concentrations. Pichia kudriavzevii produced about 35 and 200% more ethanol than the conventional Saccharomyces cerevisiae cells at 40 and 45°C, respectively. About 94% glucan in alkali-treated rice straw was converted to glucose through enzymatic hydrolysis within 36 h. Ethanol concentration of 24.25 g l⁻¹ corresponding to 82% theoretical yield on glucan basis and ethanol productivity of 1.10 g l⁻¹ h⁻¹ achieved using P. kudriavzevii during SSF hold promise for scale-up studies. An insignificant amount of glycerol and no xylitol was produced during SSF. To the best of our knowledge, this is the first study reporting ethanol production from any lignocellulosic biomass using P. kudriavzevii.

  19. Direct In Situ Viability Assessment of Bacteria in Probiotic Dairy Products Using Viability Staining in Conjunction with Confocal Scanning Laser Microscopy

    Science.gov (United States)

    Auty, M. A. E.; Gardiner, G. E.; McBrearty, S. J.; O'Sullivan, E. O.; Mulvihill, D. M.; Collins, J. K.; Fitzgerald, G. F.; Stanton, C.; Ross, R. P.

    2001-01-01

    The viability of the human probiotic strains Lactobacillus paracasei NFBC 338 and Bifidobacterium sp. strain UCC 35612 in reconstituted skim milk was assessed by confocal scanning laser microscopy using the LIVE/DEAD BacLight viability stain. The technique was rapid (diluent. PMID:11133474

  20. Evaluation of goat milk as storage media to preserve viability of human periodontal ligament cells in vitro.

    Science.gov (United States)

    Ulusoy, Ayça Tuba; Kalyoncuoglu, Elif; Kaya, Senay; Cehreli, Zafer Cavit

    2016-08-01

    The purpose of this study was to evaluate the effectiveness of goat milk as a storage media for maintenance of periodontal ligament (PDL) cell viability of avulsed teeth and compare it with commonly used and/or investigated storage media. PDL cells were obtained from the root surface of healthy premolars and were cultured in Eagle's maintenance medium (EMM). Cell cultures were treated with the following storage media: tap water (negative control); EMM (positive control); Hank's balanced salt solution; ultra high temperature (UHT) long-shelf-life lactose-free cow milk; UHT long-shelf-life whole cow milk; UHT long-shelf-life skimmed cow milk; UHT long-shelf-life soy milk; UHT long-shelf-life goat milk, UHT long-shelf-life follow on milk with probiotic, 20% propolis, and egg white. Culture plates were incubated with experimental media at 20°C for 1, 3, 6, 12, and 24 h. PDL cell viability was assessed by tetrazolium salt-based colorimetric (MTT) assay at each test period. One-way anova was used to evaluate the effects of storage solutions at each time point, followed by post hoc Duncan's multiple comparison test (P = 0.05). A dendrogram was constructed to show the arrangement of hierarchical clustering. Goat milk displayed the highest capacity to maintain cell viability at all test intervals (P milk with the probiotic showed the lowest time-dependent PDL cell viability among all test media (P milks, HBSS performed significantly less effectively in maintaining PDL cell viability during the entire test period (P milk can be recommended as a suitable storage medium for avulsed teeth. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.