[Expression, purification and immunogenicity of human papillomavirus type 11 virus-like particles from Escherichia coli].

Science.gov (United States)

Yan, Chunyan; Li, Shaowei; Wang, Jin; Wei, Minxi; Huang, Bo; Zhuang, Yudi; Li, Zhongyi; Pan, Huirong; Zhang, Jun; Xia, Ningshao

2009-11-01

To produce human papillomavirus type 11 virus-like particles (HPV11 VLPs) from Escherichia coli and to investigate its immunogenicity and type cross neutralization nature. We expressed the major capsid protein of HPV11 (HPV11-L1) in Escherichia coli ER2566 in non fusion fashion and purified by amino sulfate precipitation, ion-exchange chromatography and hydrophobic interaction chromatography, sequentially. Then we removed the reductant DTT to have the purified HPV11-L1 self-assemble into VLPs in vitro. We investigated the morphology of these VLPs with dynamic light scattering and transmission electron microscopy. We assayed the immunogenicity of the resultant HPV11 VLPs by vaccinations on mice and evaluated by HPV6/11/16/18 pseudovirion neutralization cell models. We expressed HPV11 L1 in Escherichia coli with two forms, soluble and inclusion body. The soluble HPV11 L1 with over 95% purity can self assemble to VLPs in high efficiency. Morphologically, these VLPs were globular, homogeneous and with a diameter of - 50 nm, which is quite similar with native HPV11 virions. The half effective dosage (ED50) of HPV11 VLPs is 0.031 microg, and the maximum titer of neutralizing antibody elicited is averaged to 10(6). The cross neutralization activity (against HPV6/16/18) of the anti-HPV11 serum was found to have exact correlation to the inter-type homology in amino acid alignment. We can provide HPV11 VLPs with highly immunogenicity from prokaryote expression system, which may pave a new way for research and development of prophylactic vaccine for HPV11.

In vitro screening of probiotic properties of Saccharomyces cerevisiae var. boulardii and food-borne Saccharomyces cerevisiae strains

DEFF Research Database (Denmark)

van der Aa Kuhle, Alis; Skovgaard, Kerstin; Jespersen, Lene

2005-01-01

.6-16.8%) recorded for two isolates from blue veined cheeses. Merely 25% of the S. cerevisiae var. boulardii strains displayed good adhesive properties (16.2-28.0%). The expression of the proinflammatory cytokine IL-1α decreased strikingly in IPEC-J2 cells exposed to a Shiga-like toxin 2e producing Escherichia coli...... strain when the cells were pre- and coincubated with S. cerevisiae var. boulardii even though this yeast strain was low adhesive (5.4%), suggesting that adhesion is not a mandatory prerequisite for such a probiotic effect. A strain of S. cerevisiae isolated from West African sorghum beer exerted similar......The probiotic potential of IS Saccharomyces cerevisiae strains used for production of foods or bevel-ages or isolated from such, and eight strains of Saccharomyces cerevisiae var. boulardii, was investigated. All strains included were able to withstand pH 2.5 and 0.3% Ox-all. Adhesion...

A plug-and-play pathway refactoring workflow for natural product research in Escherichia coli and Saccharomyces cerevisiae.

Science.gov (United States)

Ren, Hengqian; Hu, Pingfan; Zhao, Huimin

2017-08-01

Pathway refactoring serves as an invaluable synthetic biology tool for natural product discovery, characterization, and engineering. However, the complicated and laborious molecular biology techniques largely hinder its application in natural product research, especially in a high-throughput manner. Here we report a plug-and-play pathway refactoring workflow for high-throughput, flexible pathway construction, and expression in both Escherichia coli and Saccharomyces cerevisiae. Biosynthetic genes were firstly cloned into pre-assembled helper plasmids with promoters and terminators, resulting in a series of expression cassettes. These expression cassettes were further assembled using Golden Gate reaction to generate fully refactored pathways. The inclusion of spacer plasmids in this system would not only increase the flexibility for refactoring pathways with different number of genes, but also facilitate gene deletion and replacement. As proof of concept, a total of 96 pathways for combinatorial carotenoid biosynthesis were built successfully. This workflow should be generally applicable to different classes of natural products produced by various organisms. Biotechnol. Bioeng. 2017;114: 1847-1854. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Molecular genetic diversity of the Saccharomyces yeasts in Taiwan: Saccharomyces arboricola, Saccharomyces cerevisiae and Saccharomyces kudriavzevii.

Science.gov (United States)

Naumov, Gennadi I; Lee, Ching-Fu; Naumova, Elena S

2013-01-01

Genetic hybridization, sequence and karyotypic analyses of natural Saccharomyces yeasts isolated in different regions of Taiwan revealed three biological species: Saccharomyces arboricola, Saccharomyces cerevisiae and Saccharomyces kudriavzevii. Intraspecies variability of the D1/D2 and ITS1 rDNA sequences was detected among S. cerevisiae and S. kudriavzevii isolates. According to molecular and genetic analyses, the cosmopolitan species S. cerevisiae and S. kudriavzevii contain local divergent populations in Taiwan, Malaysia and Japan. Six of the seven known Saccharomyces species are documented in East Asia: S. arboricola, S. bayanus, S. cerevisiae, S. kudriavzevii, S. mikatae, and S. paradoxus.

L-histidine inhibits biofilm formation and FLO11-associated phenotypes in Saccharomyces cerevisiae flor yeasts.

Science.gov (United States)

Bou Zeidan, Marc; Zara, Giacomo; Viti, Carlo; Decorosi, Francesca; Mannazzu, Ilaria; Budroni, Marilena; Giovannetti, Luciana; Zara, Severino

2014-01-01

Flor yeasts of Saccharomyces cerevisiae have an innate diversity of Flo11p which codes for a highly hydrophobic and anionic cell-wall glycoprotein with a fundamental role in biofilm formation. In this study, 380 nitrogen compounds were administered to three S. cerevisiae flor strains handling Flo11p alleles with different expression levels. S. cerevisiae strain S288c was used as the reference strain as it cannot produce Flo11p. The flor strains generally metabolized amino acids and dipeptides as the sole nitrogen source, although with some exceptions regarding L-histidine and histidine containing dipeptides. L-histidine completely inhibited growth and its effect on viability was inversely related to Flo11p expression. Accordingly, L-histidine did not affect the viability of the Δflo11 and S288c strains. Also, L-histidine dramatically decreased air-liquid biofilm formation and adhesion to polystyrene of the flor yeasts with no effect on the transcription level of the Flo11p gene. Moreover, L-histidine modified the chitin and glycans content on the cell-wall of flor yeasts. These findings reveal a novel biological activity of L-histidine in controlling the multicellular behavior of yeasts [corrected].

Saccharomyces Boulardii

Science.gov (United States)

Saccharomyces boulardii is a yeast, which is a type of fungus. Saccharomyces boulardii was previously identified as a unique species of ... be a strain of Saccharomyces cerevisiae (baker's yeast). Saccharomyces boulardii is used as medicine. Saccharomyces boulardii is most ...

Fra halt ko til taberko

DEFF Research Database (Denmark)

Thomsen, Peter

2009-01-01

En halt ko kan meget nemt ende som en taberko, hvis der ikke gribes ind i tide. Ubehandlede haltheder vil ofte være  starten på en ond cirkel, som ender med, at koen bliver en taberko. Udgivelsesdato: 2009......En halt ko kan meget nemt ende som en taberko, hvis der ikke gribes ind i tide. Ubehandlede haltheder vil ofte være  starten på en ond cirkel, som ender med, at koen bliver en taberko. Udgivelsesdato: 2009...

KoFlux: Korean Regional Flux Network in AsiaFlux

Science.gov (United States)

Kim, J.

2002-12-01

AsiaFlux, the Asian arm of FLUXNET, held the Second International Workshop on Advanced Flux Network and Flux Evaluation in Jeju Island, Korea on 9-11 January 2002. In order to facilitate comprehensive Asia-wide studies of ecosystem fluxes, the meeting launched KoFlux, a new Korean regional network of long-term micrometeorological flux sites. For a successful assessment of carbon exchange between terrestrial ecosystems and the atmosphere, an accurate measurement of surface fluxes of energy and water is one of the prerequisites. During the 7th Global Energy and Water Cycle Experiment (GEWEX) Asian Monsoon Experiment (GAME) held in Nagoya, Japan on 1-2 October 2001, the Implementation Committee of the Coordinated Enhanced Observing Period (CEOP) was established. One of the immediate tasks of CEOP was and is to identify the reference sites to monitor energy and water fluxes over the Asian continent. Subsequently, to advance the regional and global network of these reference sites in the context of both FLUXNET and CEOP, the Korean flux community has re-organized the available resources to establish a new regional network, KoFlux. We have built up domestic network sites (equipped with wind profiler and radiosonde measurements) over deciduous and coniferous forests, urban and rural rice paddies and coastal farmland. As an outreach through collaborations with research groups in Japan, China and Thailand, we also proposed international flux sites at ecologically and climatologically important locations such as a prairie on the Tibetan plateau, tropical forest with mixed and rapid land use change in northern Thailand. Several sites in KoFlux already begun to accumulate interesting data and some highlights are presented at the meeting. The sciences generated by flux networks in other continents have proven the worthiness of a global array of micrometeorological flux towers. It is our intent that the launch of KoFlux would encourage other scientists to initiate and

Reconstruction of the biosynthetic pathway for the core fungal polyketide scaffold rubrofusarin in Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Rugbjerg, Peter; Naesby, Michael; Mortensen, Uffe Hasbro

2013-01-01

production in easily fermentable and genetically engineerable organisms, such as Saccharomyces cerevisiae and Escherichia coli are desirable. Rubrofusarin is an orange polyketide pigment that is a common intermediate in many different fungal biosynthetic pathways. RESULTS: In this study, we established...

Ethanol production from lignocellulosic materials. Fermentation and on-line analysis

Energy Technology Data Exchange (ETDEWEB)

Olsson, L.

1994-04-01

The fermentation performance of bacteria, yeast and fungi was investigated in lignocellulosic hydrolysates with the aim of finding microorganisms which both withstand the inhibitors and that have the ability to ferment pentoses. Firstly, the performance of Saccharomyces cidri, Saccharomyces cerevisiae, Lactobacillus brevis, Lactococcus lactis ssp lactis, Escherichia coli and Zymomonas mobilis was investigated in spent sulphite liquor and enzymatic hydrolysate of steam-pretreated willow. Secondly, the performance of natural and recombinant E. coli, Pichia stipitis, recombinant S. cerevisiae, S. cerevisiae in combination with xylose isomerase and Fusarium oxysporum was investigated in a xylose-rich acid hydrolysate of corn cob. Recombinant E. coli was the best alternative for fermentation of lignocellulosic hydrolysates, giving both high yields and productivities. The main drawback was that detoxification was necessary. The kinetics of the fermentation with recombinant E. coli KO11 was investigated in the condensate of steam-pretreated willow. A cost analysis of the ethanol production from willow was made, which predicted an ethanol production cost of 3.9 SEK/l for the pentose fermentation. The detoxification cost constituted 22% of this cost. The monitoring of three monosaccharides and ethanol in lignocellulosic hydro lysates is described. The monosaccharides were determined using immobilized pyranose oxidase in an on-line amperometric analyser. Immobilization and characterization of pyranose oxidase from Phanerochaete chrysosporium is also described. The ethanol was monitored on-line using a micro dialysis probe as an in situ sampling device. The dialysate components were then separated in a column liquid chromatographic system and the ethanol was selectively detected by an amperometric alcohol bio sensor. The determinations with on-line analysis methods agreed well with off-line methods. 248 refs, 4 figs, 12 tabs

Inactivation of Escherichia coli, Saccharomyces cerevisiae, and Lactobacillus brevis in Low-fat Milk by Pulsed Electric Field Treatment: A Pilot-scale Study.

Science.gov (United States)

Lee, Gun Joon; Han, Bok Kung; Choi, Hyuk Joon; Kang, Shin Ho; Baick, Seung Chun; Lee, Dong-Un

2015-01-01

We investigated the effects of a pulsed electric field (PEF) treatment on microbial inactivation and the physical properties of low-fat milk. Milk inoculated with Escherichia coli, Saccharomyces cerevisiae, or Lactobacillus brevis was supplied to a pilot-scale PEF treatment system at a flow rate of 30 L/h. Pulses with an electric field strength of 10 kV/cm and a pulse width of 30 μs were applied to the milk with total pulse energies of 50-250 kJ/L achieved by varying the pulse frequency. The inactivation curves of the test microorganisms were biphasic with an initial lag phase (or shoulder) followed by a phase of rapid inactivation. PEF treatments with a total pulse energy of 200 kJ/L resulted in a 4.5-log reduction in E. coli, a 4.4-log reduction in L. brevis, and a 6.0-log reduction in S. cerevisiae. Total pulse energies of 200 and 250 kJ/L resulted in greater than 5-log reductions in microbial counts in stored PEF-treated milk, and the growth of surviving microorganisms was slow during storage for 15 d at 4℃. PEF treatment did not change milk physical properties such as pH, color, or particle-size distribution (ppasteurize low-fat milk.

Dyrtid (II): Hvordan vasker man bedst en ko?

DEFF Research Database (Denmark)

Gjerløff, Anne Katrine

2008-01-01

bagsideartikel om danske landmænds problemer med effektive ko-vaskemidler i 1800-tallets sidste halvdel.......bagsideartikel om danske landmænds problemer med effektive ko-vaskemidler i 1800-tallets sidste halvdel....

Inactivation of Escherichia coli, Saccharomyces cerevisiae, and Lactobacillus brevis in Low-fat Milk by Pulsed Electric Field Treatment: A Pilot-scale Study

Science.gov (United States)

Han, Bok Kung; Choi, Hyuk Joon; Kang, Shin Ho; Baick, Seung Chun

2015-01-01

We investigated the effects of a pulsed electric field (PEF) treatment on microbial inactivation and the physical properties of low-fat milk. Milk inoculated with Escherichia coli, Saccharomyces cerevisiae, or Lactobacillus brevis was supplied to a pilot-scale PEF treatment system at a flow rate of 30 L/h. Pulses with an electric field strength of 10 kV/cm and a pulse width of 30 μs were applied to the milk with total pulse energies of 50-250 kJ/L achieved by varying the pulse frequency. The inactivation curves of the test microorganisms were biphasic with an initial lag phase (or shoulder) followed by a phase of rapid inactivation. PEF treatments with a total pulse energy of 200 kJ/L resulted in a 4.5-log reduction in E. coli, a 4.4-log reduction in L. brevis, and a 6.0-log reduction in S. cerevisiae. Total pulse energies of 200 and 250 kJ/L resulted in greater than 5-log reductions in microbial counts in stored PEF-treated milk, and the growth of surviving microorganisms was slow during storage for 15 d at 4℃. PEF treatment did not change milk physical properties such as pH, color, or particle-size distribution (pelectric-field strength of 10 kV/cm can be used to pasteurize low-fat milk. PMID:26877640

The first Koç Han: Pioneering modern architecture in Ankara

Directory of Open Access Journals (Sweden)

Oya Atalay Franck

2013-01-01

Full Text Available Koç Holding is the most established conglomerate in Turkey. Th e company’s beginnings date from the mid 1920s, when Ahmet Vehbi Koç, founder of Koç Holding, had his first own company registered with the Ankara Chamber of Commerce. In 1932, Koç moved his business from his father’s store on Anafartalar Street to a new building, the first Koç Han, erected the same year on Çankırı Street Nr.13 in Ulus. Th e building’s designer was Swiss-Austrian architect Ernst Arnold Egli, who had come to Turkey only five years earlier upon invitation by the government to work as chief architect of the Ministry of National Education. During his stay, which lasted from 1927 until 1940, Egli realized about 40 projects and worked on many more. Koç Han is the rare example of a commercial building by Egli. At a time when the urban aspect of the old town center of Ankara consisted mostly of one-or two-storeyed stone-and-wood houses, the first Koç Han represented an altogether new building type for the town, in being a multipurpose edifice with space for shops at street level, with large storage facilities below ground, and providing off ices and housing on the upper fl oors. Th e functional rigor of the plan and the sober modernism of the building’s facades contrasted strongly with the architecture of the time. In this respect, the first Koç Han was a strong statement regarding Ahmet Vehbi Koç’s belief in a modern Turkey and to the contribution of the building’s architect, Ernst Arnold Egli, to this project.

Infection with koala retrovirus subgroup B (KoRV-B), but not KoRV-A, is associated with chlamydial disease in free-ranging koalas (Phascolarctos cinereus).

Science.gov (United States)

Waugh, Courtney A; Hanger, Jonathan; Loader, Joanne; King, Andrew; Hobbs, Matthew; Johnson, Rebecca; Timms, Peter

2017-03-09

The virulence of chlamydial infection in wild koalas is highly variable between individuals. Some koalas can be infected (PCR positive) with Chlamydia for long periods but remain asymptomatic, whereas others develop clinical disease. Chlamydia in the koala has traditionally been studied without regard to coinfection with other pathogens, although koalas are usually subject to infection with koala retrovirus (KoRV). Retroviruses can be immunosuppressive, and there is evidence of an immunosuppressive effect of KoRV in vitro. Originally thought to be a single endogenous strain, a new, potentially more virulent exogenous variant (KoRV-B) was recently reported. We hypothesized that KoRV-B might significantly alter chlamydial disease outcomes in koalas, presumably via immunosuppression. By studying sub-groups of Chlamydia and KoRV infected koalas in the wild, we found that neither total KoRV load (either viraemia or proviral copies per genome), nor chlamydial infection level or strain type, was significantly associated with chlamydial disease risk. However, PCR positivity with KoRV-B was significantly associated with chlamydial disease in koalas (p = 0.02961). This represents an example of a recently evolved virus variant that may be predisposing its host (the koala) to overt clinical disease when co-infected with an otherwise asymptomatic bacterial pathogen (Chlamydia).

FLO11 gene length and transcriptional level affect biofilm-forming ability of wild flor strains of Saccharomyces cerevisiae.

Science.gov (United States)

Zara, Giacomo; Zara, Severino; Pinna, Claudia; Marceddu, Salvatore; Budroni, Marilena

2009-12-01

In Saccharomyces cerevisiae, FLO11 encodes an adhesin that is associated with different phenotypes, such as adherence to solid surfaces, hydrophobicity, mat and air-liquid biofilm formation. In the present study, we analysed FLO11 allelic polymorphisms and FLO11-associated phenotypes of 20 flor strains. We identified 13 alleles of different lengths, varying from 3.0 to 6.1 kb, thus demonstrating that FLO11 is highly polymorphic. Two alleles of 3.1 and 5.0 kb were cloned into strain BY4742 to compare the FLO11-associated phenotypes in the same genetic background. We show that there is a significant correlation between biofilm-forming ability and FLO11 length both in different and in the same genetic backgrounds. Moreover, we propose a multiple regression model that allows prediction of air-liquid biofilm-forming ability on the basis of transcription levels and lengths of FLO11 alleles in a population of S. cerevisiae flor strains. Considering that transcriptional differences are only partially explained by the differences in the promoter sequences, our results are consistent with the hypothesis that FLO11 transcription levels are strongly influenced by genetic background and affect biofilm-forming ability.

FLO11 expression and lipid biosynthesis are required for air-liquid biofilm formation in a Saccharomyces cerevisiae flor strain.

Science.gov (United States)

Zara, Giacomo; Goffrini, Paola; Lodi, Tiziana; Zara, Severino; Mannazzu, Ilaria; Budroni, Marilena

2012-11-01

Air-liquid biofilm formation is largely dependent on Flo11p and seems related to cell lipid content and composition. Here, it is shown that in the presence of cerulenin, a known inhibitor of the fatty acid synthase complex, biofilm formation is inhibited together with FLO11 transcription in a flor strain of Saccharomyces cerevisiae, while the administration of saturated fatty acids to cerulenin-containing medium restores biofilm formation and FLO11 transcription. It is also shown that, in biofilm cells, the FLO11 transcription is accompanied by the transcription of ACC1, ACS1 and INO1 key genes in lipid biosynthesis and that biofilm formation is affected by the lack of inositol in flor medium. These results are compatible with the hypothesis that the air-liquid biofilm formation depends on FLO11 transcription levels as well as on fatty acids biosynthesis. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

How Escherichia coli and Saccharomyces cerevisiae build Fe/S proteins.

Science.gov (United States)

Barras, Frédéric; Loiseau, Laurent; Py, Béatrice

2005-01-01

, plants and parasites. ISC and SUF systems share a common core function made of a cysteine desulfurase, which acts as a sulfur donor, and scaffold proteins, which act as sulfur and iron acceptors. The ISC and SUF systems also exhibit important differences. In particular, the ISC system includes an Hsp70/Hsp40-like pair of chaperones, while the SUF system involves an unorthodox ATP-binding cassette (ABC)-like component. The role of these two sets of ATP-hydrolyzing proteins in Fe/S cluster biogenesis remains unclear. Both systems are likely to target overlapping sets of apoproteins. However, regulation and phenotypic studies in E. coli, which synthesizes both types of systems, leads us to envisage ISC as the house-keeping one that functions under normal laboratory conditions, while the SUF system appears to be required in harsh environmental conditions such as oxidative stress and iron starvation. In Saccharomyces cerevisiae, the ISC system is located in the mitochondria and its function is necessary for maturation of both mitochondrial and cytosolic Fe/S proteins. Here, we attempt to provide the first comprehensive review of the ISC and SUF systems since their discovery in the mid and late 1990s. Most emphasis is put on E. coli and S. cerevisiae models with reference to other organisms when their analysis provided us with information of particular significance. We aim at covering information made available on each Isc and Suf component by the different experimental approaches, including physiology, gene regulation, genetics, enzymology, biophysics and structural biology. It is our hope that this parallel coverage will facilitate the identification of both similarities and specificities of ISC and SUF systems.

[Protective effect of Saccharomyces boulardii against intestinal mucosal barrier injury in rats with nonalcoholic fatty liver disease].

Science.gov (United States)

Liu, Y T; Li, Y Q; Wang, Y Z

2016-12-20

Objective: To investigate the protective effect of Saccharomyces boulardii against intestinal mucosal barrier injury in rats with nonalcoholic fatty liver disease (NAFLD). Methods: A total of 36 healthy male Sprague-Dawley rats with a mean body weight of 180±20 g were randomly divided into control group, model group, and treatment group, with 12 rats in each group, after adaptive feeding for 1 week. The rats in the control group were given basic feed, and those in the model group and treatment group were given high-fat feed. After 12 weeks of feeding, the treatment group was given Saccharomyces boulardii (75×10 8 CFU/kg/d) by gavage, and those in the control group and model group were given isotonic saline by gavage. At the 20th week, blood samples were taken from the abdominal aorta to measure the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), intestinal fatty acid binding protein (IFABP), tumor necrosis factor-α (TNF-α), and endotoxins. The liver pathological changes, intestinal histopathological changes, and expression of occludin in the intestinal mucosa were observed. Fecal samples were collected to measure the changes in Escherichia coli and Bacteroides. A one-way analysis of variance and the SNK test were used for comparison between multiple groups, and the rank sum test was used as the non-parametric test. Results: Compared with the control group, the model group had significantly higher body weight, liver mass, and liver index ( P 0.05). Compared with the control group, the model group had significant increases in the levels of endotoxin, TNF-α, and IFABP ( P Saccharomyces boulardii can reduce body weight and improve hepatocyte steatosis. Saccharomyces boulardii can reduce endotoxemia in NAFLD rats and thus alleviate inflammatory response. Saccharomyces boulardii can also adjust the proportion of Escherichia coli and Bacteroides in the intestine of NAFLD rats.

Population trends of Goričko agricultural landscape birds

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Denac Katarina

2017-12-01

Full Text Available Due to numerous bird surveys in the past 20 years, the avifauna of Goričko is relatively well known. For some species, the very first national ecological researches were conducted in this area. The article summarizes all bird surveys so far. It presents population trends of farmland species which is one of the most threatened bird groups in Europe. Most of the qualifying species of this habitat that are protected within the Natura 2000 network have suffered a decline at Goričko, specifically Quail Coturnix coturnix, Scops Owl Otus scops, Hoopoe Upupa epops, Woodlark Lullula arborea and White Stork Ciconia ciconia. The number of breeding pairs of the latter has not changed, but its fecundity has decreased. Furthermore, populations of other farmland bird species have decreased, for example Skylark Alauda arvensis, Stonechat Saxicola rubicola, Serin Serinus serinus and Common Linnet Linaria cannabina, as well as butterfly populations and tracts of grassland habitat types. National agricultural and nature conservation policies are evidently inefficient in protecting the biodiversity of Goričko. The most probable cause for bird population decline is agricultural intensification, which manifests itself at Goričko as disappearance and intensification of meadows, land consolidation, degradation of traditional orchards and use of pesticides. As a result of land consolidation hedges, uncultivated strips between fields, individual trees and bushes and minority habitat types are disappearing, whereas the surface of arable fields is increasing. Nature conservation measures performed by the Public Institute Goričko Nature Park with the support of DOPPS – BirdLife Slovenia volunteers seem to be efficient, but are spatially and temporally constrained. For this reason, they cannot serve as a substitute for insufficient systemic financing which could be improved by substantive and financial reform of the agri-environmental scheme. Currently, a negligible

Thermal resistance of Saccharomyces yeast ascospores in beers.

Science.gov (United States)

Milani, Elham A; Gardner, Richard C; Silva, Filipa V M

2015-08-03

The industrial production of beer ends with a process of thermal pasteurization. Saccharomyces cerevisiae and Saccharomyces pastorianus are yeasts used to produce top and bottom fermenting beers, respectively. In this research, first the sporulation rate of 12 Saccharomyces strains was studied. Then, the thermal resistance of ascospores of three S. cerevisiae strains (DSMZ 1848, DSMZ 70487, Ethanol Red(®)) and one strain of S. pastorianus (ATCC 9080) was determined in 4% (v/v) ethanol lager beer. D60 °C-values of 11.2, 7.5, 4.6, and 6.0 min and z-values of 11.7, 14.3, 12.4, and 12.7 °C were determined for DSMZ 1848, DSMZ 70487, ATCC 9080, and Ethanol Red(®), respectively. Lastly, experiments with 0 and 7% (v/v) beers were carried out to investigate the effect of ethanol content on the thermal resistance of S. cerevisiae (DSMZ 1848). D55 °C-values of 34.2 and 15.3 min were obtained for 0 and 7% beers, respectively, indicating lower thermal resistance in the more alcoholic beer. These results demonstrate similar spore thermal resistance for different Saccharomyces strains and will assist in the design of appropriate thermal pasteurization conditions for preserving beers with different alcohol contents. Copyright © 2015 Elsevier B.V. All rights reserved.

Sarcocystis neurona infection in gamma interferon gene knockout (KO) mice: comparative infectivity of sporocysts in two strains of KO mice, effect of trypsin digestion on merozoite viability, and infectivity of bradyzoites to KO mice and cell culture.

Science.gov (United States)

Dubey, J P; Sundar, N; Kwok, O C H; Saville, W J A

2013-09-01

The protozoan Sarcocystis neurona is the primary cause of Equine Protozoal Myeloencephalitis (EPM). EPM or EPM-like illness has been reported in horses, sea otters, and several other mammals. The gamma interferon gene knockout (KO) mouse is often used as a model to study biology and discovery of new therapies against S. neurona because it is difficult to induce clinical EPM in other hosts, including horses. In the present study, infectivity of three life cycle stages (merozoites, bradyzoites, sporozoites) to KO mice and cell culture was studied. Two strains of KO mice (C57-black, and BALB/c-derived, referred here as black or white) were inoculated orally graded doses of S. neurona sporocysts; 12 sporocysts were infective to both strains of mice and all infected mice died or became ill within 70 days post-inoculation. Although there was no difference in infectivity of sporocysts to the two strains of KO mice, the disease was more severe in black mice. S. neurona bradyzoites were not infectious to KO mice and cell culture. S. neurona merozoites survived 120 min incubation in 0.25% trypsin, indicating that trypsin digestion can be used to recover S. neurona from tissues of acutely infected animals. Published by Elsevier B.V.

Impaired memory of eyeblink conditioning in CaMKIV KO mice.

Science.gov (United States)

Lee, Ka Hung; Chatila, Talal A; Ram, Rana A; Thompson, Richard F

2009-04-01

The calcium/calmodulin-dependent protein kinase type IV (CaMKIV) is highly expressed in cerebellar cortical granule cells and deep nuclear neurons in the cerebellum. It mediates the phosphorylation and activation of the cAMP-dependent response element binding protein (CREB). In several paradigms CREB-dependent transcription is required for cellular events underlying long-term memory processes. Also, CaMKIV deficiency results in impaired long-term depression (LTD) induction in cerebellar cortex. To investigate the function of CaMKIV in the cerebellum, Wild-type (WT) and CaMKIV KO mice were tested with delay eyeblink conditioning. KO and WT mice did not differ in acquisition, but the KO mice showed a significantly lower conditioned response (CR) percentage than the WT mice in the retention testing and retraining period. The CR peak latencies for the two groups did not differ in acquisition but were shorter for the KO mice in the testing period. No significant differences were found between KO and WT mice in spontaneous eyeblink activity, auditory brainstem response (ABR) amplitudes, and tail-flick latency. The results suggest an important role for CaMKIV in long-term memory in the cerebellum. (c) 2009 APA, all rights reserved.

Growth inhibition of S. cerevisiae, B. subtilis, and E. coli by lignocellulosic and fermentation products

NARCIS (Netherlands)

Carvalho Pereira, Joana P.C.; Verheijen, P.J.T.; Straathof, Adrie J.J.

2016-01-01

This paper describes the effect of several inhibiting components on three potential hosts for the bio-based production of methyl propionate, namely, wild-type Escherichia coli and Bacillus subtilis, and evolved Saccharomyces cerevisiae IMS0351. The inhibition by the lignocellulose-derived

Characterization of the insulin sensitivity of ghrelin receptor KO mice using glycemic clamps

Directory of Open Access Journals (Sweden)

Morgan Kristen

2011-01-01

Full Text Available Abstract Background We and others have demonstrated previously that ghrelin receptor (GhrR knock out (KO mice fed a high fat diet (HFD have increased insulin sensitivity and metabolic flexibility relative to WT littermates. A striking feature of the HFD-fed GhrR KO mouse is the dramatic decrease in hepatic steatosis. To characterize further the underlying mechanisms of glucose homeostasis in GhrR KO mice, we conducted both hyperglycemic (HG and hyperinsulinemic-euglycemic (HI-E clamps. Additionally, we investigated tissue glucose uptake and specifically examined liver insulin sensitivity. Results Consistent with glucose tolerance-test data, in HG clamp experiments, GhrR KO mice showed a reduction in glucose-stimulated insulin release relative to WT littermates. Nevertheless, a robust 1st phase insulin secretion was still achieved, indicating that a healthy β-cell response is maintained. Additionally, GhrR KO mice demonstrated both a significantly increased glucose infusion rate and significantly reduced insulin requirement for maintenance of the HG clamp, consistent with their relative insulin sensitivity. In HI-E clamps, both LFD-fed and HFD-fed GhrR KO mice showed higher peripheral insulin sensitivity relative to WT littermates as indicated by a significant increase in insulin-stimulated glucose disposal (Rd, and decreased hepatic glucose production (HGP. HFD-fed GhrR KO mice showed a marked increase in peripheral tissue glucose uptake in a variety of tissues, including skeletal muscle, brown adipose tissue and white adipose tissue. GhrR KO mice fed a HFD also showed a modest, but significant decrease in conversion of pyruvate to glucose, as would be anticipated if these mice displayed increased liver insulin sensitivity. Additionally, the levels of UCP2 and UCP1 were reduced in the liver and BAT, respectively, in GhrR KO mice relative to WT mice. Conclusions These results indicate that improved glucose homeostasis of GhrR KO mice is

One-pot green synthesis of carbon dots by using Saccharum officinarum juice for fluorescent imaging of bacteria (Escherichia coli) and yeast (Saccharomyces cerevisiae) cells

Energy Technology Data Exchange (ETDEWEB)

Mehta, Vaibhavkumar N. [Applied Chemistry Department, S. V. National Institute of Technology, Surat, 395 007 (India); Jha, Sanjay [Gujarat Agricultural Biotechnology Institute, Navsari Agricultural University, Surat, 395007 (India); Kailasa, Suresh Kumar, E-mail: sureshkumarchem@gmail.com [Applied Chemistry Department, S. V. National Institute of Technology, Surat, 395 007 (India)

2014-05-01

We are reporting highly economical plant-based hydrothermal method for one-pot green synthesis of water-dispersible fluorescent carbon dots (CDs) by using Saccharum officinarum juice as precursor. The synthesized CDs were characterized by UV-visible, fluorescence, Fourier transform infrared (FT-IR), dynamic light scattering (DLS), high-resolution transmission electron microscopic (HR-TEM), and laser scanning confocal microscopic techniques. The CDs are well dispersed in water with an average size of ∼ 3 nm and showed bright blue fluorescence under UV-light (λ{sub ex} = 365 nm). These CDs acted as excellent fluorescent probes in cellular imaging of bacteria (Escherichia coli) and yeast (Saccharomyces cerevisiae). - Highlights: • One-pot green synthesis was used for fluorescent CDs. • FT-IR, DLS, and TEM were used for the characterization of CDs. • The CDs are well dispersed in water with an average size of ∼ 3 nm. • The CDs acted as fluorescent probes for imaging of bacteria and yeast cells.

Superoxide Stabilization and a Universal KO2 Growth Mechanism in Potassium-Oxygen Batteries.

Science.gov (United States)

Wang, Wanwan; Lai, Nien-Chu; Liang, Zhuojian; Wang, Yu; Lu, Yi-Chun

2018-04-23

Rechargeable potassium-oxygen (K-O 2 ) batteries promise to provide higher round-trip efficiency and cycle life than other alkali-oxygen batteries with satisfactory gravimetric energy density (935 Wh kg -1 ). Exploiting a strong electron-donating solvent, for example, dimethyl sulfoxide (DMSO) strongly stabilizes the discharge product (KO 2 ), resulting in significant improvement in electrode kinetics and chemical/electrochemical reversibility. The first DMSO-based K-O 2 battery demonstrates a much higher energy efficiency and stability than the glyme-based electrolyte. A universal KO 2 growth model is developed and it is demonstrated that the ideal solvent for K-O 2 batteries should strongly stabilize superoxide (strong donor ability) to obtain high electrode kinetics and reversibility while providing fast oxygen diffusion to achieve high discharge capacity. This work elucidates key electrolyte properties that control the efficiency and reversibility of K-O 2 batteries. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Prevajanje športne terminologije v priročniku Osnove košarke NBA

OpenAIRE

Kajič, Matic

2016-01-01

Pri prevajanju strokovnih besedil ne gre le za vestno opravljanje veščin, ampak za visoko intelektualno delovanje: reševanje vedno novih problemov. To velja tudi za košarkarsko terminologijo, saj obstaja veliko ameriških košarkarskih izrazov, za katere nimamo ustreznic v slovenskem jeziku. Posebnost priročnika Osnove košarke NBA avtorja Marka Vancila se kaže v velikem številu strokovnega košarkarskega izrazja, ki od prevajalca zahteva veliko mero strokovnega poznavanja področja. V diplomskem ...

Metabolic regulation analysis of an ethanologenic Escherichia coli strain based on RT-PCR and enzymatic activities

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Martinez Alfredo

2008-05-01

Full Text Available Abstract Background A metabolic regulation study was performed, based upon measurements of enzymatic activities, fermentation performance, and RT-PCR analysis of pathways related to central carbon metabolism, in an ethanologenic Escherichia coli strain (CCE14 derived from lineage C. In comparison with previous engineered strains, this E coli derivative has a higher ethanol production rate in mineral medium, as a result of the elevated heterologous expression of the chromosomally integrated genes encoding PDCZm and ADHZm (pyruvate decarboxylase and alcohol dehydrogenase from Zymomonas mobilis. It is suggested that this behavior might be due to lineage differences between E. coli W and C. Results This study demonstrated that the glycolytic flux is controlled, in this case, by reactions outside glycolysis, i.e., the fermentative pathways. Changes in ethanol production rate in this ethanologenic strain result in low organic acid production rates, and high glycolytic and ethanologenic fluxes, that correlate with enhanced transcription and enzymatic activity levels of PDCZm and ADHZm. Furthermore, a higher ethanol yield (90% of the theoretical in glucose-mineral media was obtained with CCE14 in comparison with previous engineered E. coli strains, such as KO11, that produces a 70% yield under the same conditions. Conclusion Results suggest that a higher ethanol formation rate, caused by ahigher PDCZm and ADHZm activities induces a metabolic state that cells compensate through enhanced glucose transport, ATP synthesis, and NAD-NADH+H turnover rates. These results show that glycolytic enzymatic activities, present in E. coli W and C under fermentative conditions, are sufficient to contend with increases in glucose consumption and product formation rates.

In vitro screening of probiotic properties of Saccharomyces cerevisiae var. boulardii and food-borne Saccharomyces cerevisiae strains.

Science.gov (United States)

van der Aa Kühle, Alis; Skovgaard, Kerstin; Jespersen, Lene

2005-05-01

The probiotic potential of 18 Saccharomyces cerevisiae strains used for production of foods or beverages or isolated from such, and eight strains of Saccharomyces cerevisiae var. boulardii, was investigated. All strains included were able to withstand pH 2.5 and 0.3% Oxgall. Adhesion to the nontumorigenic porcine jejunal epithelial cell line (IPEC-J2) was investigated by incorporation of 3H-methionine into the yeast cells and use of liquid scintillation counting. Only few of the food-borne S. cerevisiae strains exhibited noteworthy adhesiveness with the strongest levels of adhesion (13.6-16.8%) recorded for two isolates from blue veined cheeses. Merely 25% of the S. cerevisiae var. boulardii strains displayed good adhesive properties (16.2-28.0%). The expression of the proinflammatory cytokine IL-1alpha decreased strikingly in IPEC-J2 cells exposed to a Shiga-like toxin 2e producing Escherichia coli strain when the cells were pre- and coincubated with S. cerevisiae var. boulardii even though this yeast strain was low adhesive (5.4%), suggesting that adhesion is not a mandatory prerequisite for such a probiotic effect. A strain of S. cerevisiae isolated from West African sorghum beer exerted similar effects hence indicating that food-borne strains of S. cerevisiae may possess probiotic properties in spite of low adhesiveness.

Amino acid transporter genes are essential for FLO11-dependent and FLO11-independent biofilm formation and invasive growth in Saccharomyces cerevisiae.

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Rasmus Torbensen

Full Text Available Amino acids can induce yeast cell adhesion but how amino acids are sensed and signal the modulation of the FLO adhesion genes is not clear. We discovered that the budding yeast Saccharomyces cerevisiae CEN.PK evolved invasive growth ability under prolonged nitrogen limitation. Such invasive mutants were used to identify amino acid transporters as regulators of FLO11 and invasive growth. One invasive mutant had elevated levels of FLO11 mRNA and a Q320STOP mutation in the SFL1 gene that encodes a protein kinase A pathway regulated repressor of FLO11. Glutamine-transporter genes DIP5 and GNP1 were essential for FLO11 expression, invasive growth and biofilm formation in this mutant. Invasive growth relied on known regulators of FLO11 and the Ssy1-Ptr3-Ssy5 complex that controls DIP5 and GNP1, suggesting that Dip5 and Gnp1 operates downstream of the Ssy1-Ptr3-Ssy5 complex for regulation of FLO11 expression in a protein kinase A dependent manner. The role of Dip5 and Gnp1 appears to be conserved in the S. cerevisiae strain ∑1278b since the dip5 gnp1 ∑1278b mutant showed no invasive phenotype. Secondly, the amino acid transporter gene GAP1 was found to influence invasive growth through FLO11 as well as other FLO genes. Cells carrying a dominant loss-of-function PTR3(647::CWNKNPLSSIN allele had increased transcription of the adhesion genes FLO1, 5, 9, 10, 11 and the amino acid transporter gene GAP1. Deletion of GAP1 caused loss of FLO11 expression and invasive growth. However, deletions of FLO11 and genes encoding components of the mitogen-activated protein kinase pathway or the protein kinase A pathway were not sufficient to abolish invasive growth, suggesting involvement of other FLO genes and alternative pathways. Increased intracellular amino acid pools in the PTR3(647::CWNKNPLSSIN-containing strain opens the possibility that Gap1 regulates the FLO genes through alteration of the amino acid pool sizes.

The Mobilome; A Major Contributor to Escherichia coli stx2-Positive O26:H11 Strains Intra-Serotype Diversity

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Sabine Delannoy

2017-09-01

Full Text Available Shiga toxin-producing Escherichia coli of serotype O26:H11/H- constitute a diverse group of strains and several clones with distinct genetic characteristics have been identified and characterized. Whole genome sequencing was performed using Illumina and PacBio technologies on eight stx2-positive O26:H11 strains circulating in France. Comparative analyses of the whole genome of the stx2-positive O26:H11 strains indicate that several clones of EHEC O26:H11 are co-circulating in France. Phylogenetic analysis of the French strains together with stx2-positive and stx-negative E. coli O26:H11 genomes obtained from Genbank indicates the existence of four clonal complexes (SNP-CCs separated in two distinct lineages, one of which comprises the “new French clone” (SNP-CC1 that appears genetically closely related to stx-negative attaching and effacing E. coli (AEEC strains. Interestingly, the whole genome SNP (wgSNP phylogeny is summarized in the cas gene phylogeny, and a simple qPCR assay targeting the CRISPR array specific to SNP-CC1 (SP_O26-E can distinguish between the two main lineages. The PacBio sequencing allowed a detailed analysis of the mobile genetic elements (MGEs of the strains. Numerous MGEs were identified in each strain, including a large number of prophages and up to four large plasmids, representing overall 8.7–19.8% of the total genome size. Analysis of the prophage pool of the strains shows a considerable diversity with a complex history of recombination. Each clonal complex (SNP-CC is characterized by a unique set of plasmids and phages, including stx-prophages, suggesting evolution through separate acquisition events. Overall, the MGEs appear to play a major role in O26:H11 intra-serotype clonal diversification.

The Mobilome; A Major Contributor to Escherichia coli stx2-Positive O26:H11 Strains Intra-Serotype Diversity.

Science.gov (United States)

Delannoy, Sabine; Mariani-Kurkdjian, Patricia; Webb, Hattie E; Bonacorsi, Stephane; Fach, Patrick

2017-01-01

Shiga toxin-producing Escherichia coli of serotype O26:H11/H- constitute a diverse group of strains and several clones with distinct genetic characteristics have been identified and characterized. Whole genome sequencing was performed using Illumina and PacBio technologies on eight stx2 -positive O26:H11 strains circulating in France. Comparative analyses of the whole genome of the stx2 -positive O26:H11 strains indicate that several clones of EHEC O26:H11 are co-circulating in France. Phylogenetic analysis of the French strains together with stx2 -positive and stx -negative E. coli O26:H11 genomes obtained from Genbank indicates the existence of four clonal complexes (SNP-CCs) separated in two distinct lineages, one of which comprises the "new French clone" (SNP-CC1) that appears genetically closely related to stx -negative attaching and effacing E. coli (AEEC) strains. Interestingly, the whole genome SNP (wgSNP) phylogeny is summarized in the cas gene phylogeny, and a simple qPCR assay targeting the CRISPR array specific to SNP-CC1 (SP_O26-E) can distinguish between the two main lineages. The PacBio sequencing allowed a detailed analysis of the mobile genetic elements (MGEs) of the strains. Numerous MGEs were identified in each strain, including a large number of prophages and up to four large plasmids, representing overall 8.7-19.8% of the total genome size. Analysis of the prophage pool of the strains shows a considerable diversity with a complex history of recombination. Each clonal complex (SNP-CC) is characterized by a unique set of plasmids and phages, including stx -prophages, suggesting evolution through separate acquisition events. Overall, the MGEs appear to play a major role in O26:H11 intra-serotype clonal diversification.

Karyotypes of Saccharomyces sensu lato species

DEFF Research Database (Denmark)

Petersen, Randi Føns; Nilsson-Tilgren, Torsten; Piskur, Jure

1999-01-01

An improved pulsed-field electrophoresis program was developed to study differently sized chromosomes within the genus Saccharomyces. The number of chromosomes in the type strains was shown to be nine in Saccharomyces castellii and Saccharomyces dairenensis, 12 in Saccharomyces servazzii...... and Saccharomyces unisporus, 16 in Saccharomyces exiguus and seven in Saccharomyces kluyveri. The sizes of individual chromosomes were resolved and the approximate genome sizes were determined by the addition of individual chromosomes of the karyotypes. Apparently. the genome of S. exiguus, which is the only...... Saccharomyces sensu late yeast to contain small chromosomes, is larger than that of Saccharomyces cerevisiae. On the other hand, other species exhibited genome sizes that were 10-25% smaller than that of S. cerevisiae. Well-defined karyotypes represent the basis for future genome mapping and sequencing projects...

Ethanol production from Jerusalem artichoke by strains of Saccharomyces cheresiensis and Saccharomyces beticus

Energy Technology Data Exchange (ETDEWEB)

Pourrat, H.; Barthomeuf, C.; Regerat, F.; Carnat, A.P.; Carnat, A.

1983-03-01

Ethanol production from Jerusalem artichoke which is the most interesting autochtonous material has been studied. Two selected and acclimatised strains of Saccharomyces: Saccharomyces cheresiensis and Saccharomyces beticus were retained. The fermentation conditions, exactly definited, makes it possible to obtain in 4 days a theoric yield.

Effect of menadione and hydrogen peroxide on catalase activity in Saccharomyces yeast strains

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Nadejda EFREMOVA

2013-05-01

Full Text Available It has been studied the possibility of utilization of two important oxidant factors as regulators of catalase activity in Saccharomyces yeasts. In this paper results of the screening of some Saccharomyces yeast strains for potential producers of catalase are presented. Results of the screening for potential catalase producer have revealed that Saccharomyces cerevisiae CNMN-Y-11 strain possesses the highest catalase activity (2900 U/mg protein compared with other samples. Maximum increase of catalase activity with 50-60% compared to the reference sample was established in the case of hydrogen peroxide and menadione utilization in optimal concentrations of 15 and 10 mM. This research has been demonstrated the potential benefits of application of hydrogen peroxide and menadione as stimulatory factors of catalase activity in Saccharomyces yeasts.

Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol

Energy Technology Data Exchange (ETDEWEB)

Steen, EricJ.; Chan, Rossana; Prasad, Nilu; Myers, Samuel; Petzold, Christopher; Redding, Alyssa; Ouellet, Mario; Keasling, JayD.

2008-11-25

BackgroundIncreasing energy costs and environmental concerns have motivated engineering microbes for the production of ?second generation? biofuels that have better properties than ethanol.Results& ConclusionsSaccharomyces cerevisiae was engineered with an n-butanol biosynthetic pathway, in which isozymes from a number of different organisms (S. cerevisiae, Escherichia coli, Clostridium beijerinckii, and Ralstonia eutropha) were substituted for the Clostridial enzymes and their effect on n-butanol production was compared. By choosing the appropriate isozymes, we were able to improve production of n-butanol ten-fold to 2.5 mg/L. The most productive strains harbored the C. beijerinckii 3-hydroxybutyryl-CoA dehydrogenase, which uses NADH as a co-factor, rather than the R. eutropha isozyme, which uses NADPH, and the acetoacetyl-CoA transferase from S. cerevisiae or E. coli rather than that from R. eutropha. Surprisingly, expression of the genes encoding the butyryl-CoA dehydrogenase from C. beijerinckii (bcd and etfAB) did not improve butanol production significantly as previously reported in E. coli. Using metabolite analysis, we were able to determine which steps in the n-butanol biosynthetic pathway were the most problematic and ripe for future improvement.

Overview of the KoRIA Facility for Rare Isotope Beams

International Nuclear Information System (INIS)

Hong, S.W.; Bak, S.I.; Chai, J.S.; Ahn, J.K.; Blumenfeld, Y.; Cheon, B.-G.; Choi, C.I.; Cheoun, M.-K.; Cho, D.; Cho, Y.S.; Choi, B.H.; Choi, E.M.; And others

2013-01-01

The Korea Rare Isotope Accelerator, currently referred to as KoRIA, is briefly presented. The KoRIA facility is aimed to enable cutting-edge sciences in a wide range of fields. It consists of a 70 kW isotope separator on-line (ISOL) facility driven by a 70 MeV, 1 mA proton cyclotron and a 400 kW in-flight fragmentation (IFF) facility. The ISOL facility uses a superconducting (SC) linac for post-acceleration of rare isotopes up to about 18 MeV/u, while the SC linac of IFF facility is capable of accelerating uranium beams up to 200 MeV/u, 8 pμA and proton beams up to 600 MeV, 660 μA. Overall features of the KoRIA facility are presented with a focus on the accelerator design. (author)

Kinetics of phosphomevalonate kinase from Saccharomyces cerevisiae.

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David E Garcia

Full Text Available The mevalonate-based isoprenoid biosynthetic pathway is responsible for producing cholesterol in humans and is used commercially to produce drugs, chemicals, and fuels. Heterologous expression of this pathway in Escherichia coli has enabled high-level production of the antimalarial drug artemisinin and the proposed biofuel bisabolane. Understanding the kinetics of the enzymes in the biosynthetic pathway is critical to optimize the pathway for high flux. We have characterized the kinetic parameters of phosphomevalonate kinase (PMK, EC 2.7.4.2 from Saccharomyces cerevisiae, a previously unstudied enzyme. An E. coli codon-optimized version of the S. cerevisiae gene was cloned into pET-52b+, then the C-terminal 6X His-tagged protein was expressed in E. coli BL21(DE3 and purified on a Ni²⁺ column. The KM of the ATP binding site was determined to be 98.3 µM at 30°C, the optimal growth temperature for S. cerevisiae, and 74.3 µM at 37°C, the optimal growth temperature for E. coli. The K(M of the mevalonate-5-phosphate binding site was determined to be 885 µM at 30°C and 880 µM at 37°C. The V(max was determined to be 4.51 µmol/min/mg enzyme at 30°C and 5.33 µmol/min/mg enzyme at 37°C. PMK is Mg²⁺ dependent, with maximal activity achieved at concentrations of 10 mM or greater. Maximum activity was observed at pH = 7.2. PMK was not found to be substrate inhibited, nor feedback inhibited by FPP at concentrations up to 10 µM FPP.

Oestrogen-deficient female aromatase knockout (ArKO) mice exhibit depressive-like symptomatology.

Science.gov (United States)

Dalla, C; Antoniou, K; Papadopoulou-Daifoti, Z; Balthazart, J; Bakker, J

2004-07-01

We recently found that female aromatase knockout (ArKO) mice that are deficient in oestradiol due to a targeted mutation in the aromatase gene show deficits in sexual behaviour that cannot be corrected by adult treatment with oestrogens. We determined here whether these impairments are associated with changes in general levels of activity, anxiety or 'depressive-like' symptomatology due to chronic oestrogen deficiency. We also compared the neurochemical profile of ArKO and wild-type (WT) females, as oestrogens have been shown to modulate dopaminergic, serotonergic and noradrenergic brain activities. ArKO females did not differ from WT in spontaneous motor activity, exploration or anxiety. These findings are in line with the absence of major neurochemical alterations in hypothalamus, prefrontal cortex or striatum, which are involved in the expression of these behaviours. By contrast, ArKO females displayed decreased active behaviours, such as struggling and swimming, and increased passive behaviours, such as floating, in repeated sessions of the forced swim test, indicating that these females exhibit 'depressive-like' symptoms. Adult treatment with oestradiol did not reverse the behavioural deficits observed in the forced swim test, suggesting that they may be due to the absence of oestradiol during development. Accordingly, an increased serotonergic activity was observed in the hippocampus of ArKO females compared with WT, which was also not reversed by adult oestradiol treatment. The possible organizational role of oestradiol on the hippocampal serotonergic system and the 'depressive-like' profile of ArKO females provide new insights into the pathophysiology of depression and the increased vulnerability of women to depression.

Repair of UV-irradiated plasmid DNA in excision repair deficient mutants of Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Ikai, K.; Tano, K.; Ohnishi, T.; Nozu, K.

1985-01-01

The repair of UV-irradiated DNA of plasmid YEp13 was studied in the incision defective strains by measurement of cell transformation frequency. In Saccharomyces cerevisiae, rad1,2,3 and 4 mutants could repair UV-damaged plasmid DNA. In Escherichia coli, uvrA mutant was unable to repair UV-damaged plasmid DNA; however, pretreatment of the plasmid with Micrococcus luteus endonuclease increased repair. It was concluded that all the mutations of yeast were probably limited only to the nuclear DNA. (author)

Positron Emission Tomographic Imaging of the Cannabinoid Type 1 Receptor System with [11C]OMAR ([11C]JHU75528: Improvements in Image Quantification Using Wild-Type and Knockout Mice

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Raúl Herance

2011-11-01

Full Text Available In this study, we assessed the feasibility of using positron emission tomography (PET and the tracer [11C]OMAR ([11C]JHU75528, an analogue of rimonabant, to study the brain cannabinoid type 1 (CB1 receptor system. Wild-type (WT andCB1 knockout (KO animals were imaged at baseline and after pretreatment with blocking doses of rimonabant. Brain uptake in WT animals was higher (50% than in KO animals in baseline conditions. After pretreatment with rimonabant, WT uptake lowered to the level of KO animals. The results of this study support the feasibility of using PET with the radiotracer [11C]JHU75528 to image the brain CB1 receptor system in mice. In addition, this methodology can be used to assess the effect of new drugs in preclinical studies using genetically manipulated animals.

Construction of a novel kind of expression plasmid by homologous recombination in Saccharomyces cerevisiae

Institute of Scientific and Technical Information of China (English)

CHEN; Xiangling

2005-01-01

[1]Brunelli, J. P., Pall, M. L., A series of yeast vectors for expression of cDNAs and other DNA sequences, Yeast, 1993, 9: 1299―1308.[2]Sikorski, R. S., Hieter, P., A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae, Genetics, 1989, 122: 19―27.[3]Bonneaud, N., Ozier-Kalogerogoulos, O., Li, G. et al., A family of low and high copy replicative, integrative and single-stranded S. cerevisiae /E. coli shuttle vector, Yeast, 1991, 7: 609―615.[4]Huo, K. K., Yu, L. L., Chen, X. J., Li, Y. Y., A stable vector for high-level expression and secretion of human interferon alpha A in yeast, Science in China, Ser. B, 1993, 36(5): 557―567.[5]Zhou, Z. X., Yuan, H. Y., He, W. et al., Expression of the modified HBsAg gene SA-28 directed by a constitutive promoter, Journal of Fudan university (Natural Science), 2000, 39(3): 264―268.[6]Paques, F., Haber, J. E., Multiple pathways of recombination induces by double-strand breaks in Saccharomyces cerevisiae, Microbiology and Molecular Biology Reviews, 1999, 63(2): 349―404.[7]Martin, K., Damage-induced recombination in the yeast Saccharomyces cerevisiae, Mutation Research, 2000, 451: 91―105.[8]Alira, S., Tomoko, O., Homologous recombination and the roles of double-strand breaks, TIBS, 1995, 20: 387―391.[9]Patrick, S., Kelly, M. T., Stephen, V. K., Recombination factor of Saccharomyces cerevisiae, Mutation Research, 2000, 451: 257―275.[10]Manivasakam, P., Weber, S. C., McElver, J., Schiestl, R. H., Micro-homology mediated PCR targeting in Saccharomyces cerevisiae, Nucleic Acids Res., 1995, 23(14): 2799―2800.[11]Baudin, A., Lacroute, F., Cullin, C., A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae, Nucleic Acids Res., 1993, 21(14): 3329―3330.[12]Hua, S. B., Qiu, M., Chan, E., Zhu, L., Luo, Y., Minimum length of sequence homology required for in vivo cloning by homolo-gous recombination in yeast, Plasmid, 1997, 38

The Saccharomyces Genome Database Variant Viewer.

Science.gov (United States)

Sheppard, Travis K; Hitz, Benjamin C; Engel, Stacia R; Song, Giltae; Balakrishnan, Rama; Binkley, Gail; Costanzo, Maria C; Dalusag, Kyla S; Demeter, Janos; Hellerstedt, Sage T; Karra, Kalpana; Nash, Robert S; Paskov, Kelley M; Skrzypek, Marek S; Weng, Shuai; Wong, Edith D; Cherry, J Michael

2016-01-04

The Saccharomyces Genome Database (SGD; http://www.yeastgenome.org) is the authoritative community resource for the Saccharomyces cerevisiae reference genome sequence and its annotation. In recent years, we have moved toward increased representation of sequence variation and allelic differences within S. cerevisiae. The publication of numerous additional genomes has motivated the creation of new tools for their annotation and analysis. Here we present the Variant Viewer: a dynamic open-source web application for the visualization of genomic and proteomic differences. Multiple sequence alignments have been constructed across high quality genome sequences from 11 different S. cerevisiae strains and stored in the SGD. The alignments and summaries are encoded in JSON and used to create a two-tiered dynamic view of the budding yeast pan-genome, available at http://www.yeastgenome.org/variant-viewer. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Datengeleitetes Lernen im studienbegleitenden Deutschunterricht am Beispiel des KoGloss-Ansatzes

OpenAIRE

Dubova, Agnese; Proveja, Egita

2016-01-01

Der vorliegende Aufsatz stellt den sprachdidaktischen Ansatz KoGloss vor und beschreibt die Möglichkeiten seines Einsatzes im studienbegleitenden Deutschunterricht. Als eine der Formen des datengeleiteten Lernens ermöglicht der KoGloss-Ansatz eine forschungsorientierte und lernerzentrierte Herangehensweise, die insbesondere im akademischen Sprachunterricht gefragt ist. Eine korpusbasierte Erschließung von (Fach-)Wörtern und komplexen sprachlichen Mustern, das learning by doing, die Kooperatio...

Dose-Dependent Rescue of KO Amelogenin Enamel by Transgenes in Vivo

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Felicitas B. Bidlack

2017-11-01

Full Text Available Mice lacking amelogenin (KO have hypoplastic enamel. Overexpression of the most abundant amelogenin splice variant M180 and LRAP transgenes can substantially improve KO enamel, but only ~40% of the incisor thickness is recovered and the prisms are not as tightly woven as in WT enamel. This implies that the compositional complexity of the enamel matrix is required for different aspects of enamel formation, such as organizational structure and thickness. The question arises, therefore, how important the ratio of different matrix components, and in particular amelogenin splice products, is in enamel formation. Can optimal expression levels of amelogenin transgenes representing both the most abundant splice variants and cleavage product at protein levels similar to that of WT improve the enamel phenotype of KO mice? Addressing this question, our objective was here to understand dosage effects of amelogenin transgenes (Tg representing the major splice variants M180 and LRAP and cleavage product CTRNC on enamel properties. Amelogenin KO mice were mated with M180Tg, CTRNCTg and LRAPTg mice to generate M180Tg and CTRNCTg double transgene and M180Tg, CTRNCTg, LRAPTg triple transgene mice with transgene hemizygosity (on one allelle or homozygosity (on both alleles. Transgene homo- vs. hemizygosity was determined by qPCR and relative transgene expression confirmed by Western blot. Enamel volume and mineral density were analyzed by microCT, thickness and structure by SEM, and mechanical properties by Vickers microhardness testing. There were no differences in incisor enamel thickness between amelogenin KO mice with three or two different transgenes, but mice homozygous for a given transgene had significantly thinner enamel than mice hemizygous for the transgene (p < 0.05. The presence of the LRAPTg did not improve the phenotype of M180Tg/CTRNCTg/KO enamel. In the absence of endogenous amelogenin, the addition of amelogenin transgenes representing the most

Saccharomyces boulardii Preserves the Barrier Function and Modulates the Signal Transduction Pathway Induced in Enteropathogenic Escherichia coli-Infected T84 Cells

Science.gov (United States)

Czerucka, Dorota; Dahan, Stephanie; Mograbi, Baharia; Rossi, Bernard; Rampal, Patrick

2000-01-01

Use of the nonpathogenic yeast Saccharomyces boulardii in the treatment of infectious diarrhea has attracted growing interest. The present study designed to investigate the effect of this yeast on enteropathogenic Escherichia coli (EPEC)-associated disease demonstrates that S. boulardii abrogated the alterations induced by an EPEC strain on transepithelial resistance, [3H]inulin flux, and ZO-1 distribution in T84 cells. Moreover, EPEC-mediated apoptosis of epithelial cells was delayed in the presence of S. boulardii. The yeast did not modify the number of adherent bacteria but lowered by 50% the number of intracellular bacteria. Infection by EPEC induced tyrosine phosphorylation of several proteins in T84 cells, including p46 and p52 SHC isoforms, that was attenuated in the presence of S. boulardii. Similarly, EPEC-induced activation of the ERK1/2 mitogen-activated protein (MAP) kinase pathway was diminished in the presence of the yeast. Interestingly, inhibition of the ERK1/2 pathway with the specific inhibitor PD 98059 decreased EPEC internalization, suggesting that modulation of the ERK1/2 MAP pathway might account for the lowering of the number of intracellular bacteria observed in the presence of S. boulardii. Altogether, this study demonstrated that S. boulardii exerts a protective effect on epithelial cells after EPEC adhesion by modulating the signaling pathway induced by bacterial infection. PMID:10992512

Predicting Maps of Green Growth in Košice

Science.gov (United States)

Poorova, Zuzana; Vranayova, Zuzana

2017-10-01

The paper deals with the changing of the traditional roofs in the city of Košice into green roofs. Possible areas of city housing estates, after taking into account the conditions of each of them (types of buildings, statics of buildings), are listed in the paper. The research is picturing the prediction maps of Košice city from 2017 to 2042 in 5-years interval. The paper is a segment of a dissertation work focusing on changing traditional roofs into green roofs with the aim to retain water, calculate the amount of retained water and show possibilities how to use this water.

Diversity of Shiga Toxin-Producing Escherichia coli (STEC) O26:H11 Strains Examined via stx Subtypes and Insertion Sites of Stx and EspK Bacteriophages

Science.gov (United States)

Bonanno, Ludivine; Loukiadis, Estelle; Mariani-Kurkdjian, Patricia; Oswald, Eric; Garnier, Lucille; Michel, Valérie

2015-01-01

Shiga toxin-producing Escherichia coli (STEC) is a food-borne pathogen that may be responsible for severe human infections. Only a limited number of serotypes, including O26:H11, are involved in the majority of serious cases and outbreaks. The main virulence factors, Shiga toxins (Stx), are encoded by bacteriophages. Seventy-four STEC O26:H11 strains of various origins (including human, dairy, and cattle) were characterized for their stx subtypes and Stx phage chromosomal insertion sites. The majority of food and cattle strains possessed the stx1a subtype, while human strains carried mainly stx1a or stx2a. The wrbA and yehV genes were the main Stx phage insertion sites in STEC O26:H11, followed distantly by yecE and sbcB. Interestingly, the occurrence of Stx phages inserted in the yecE gene was low in dairy strains. In most of the 29 stx-negative E. coli O26:H11 strains also studied here, these bacterial insertion sites were vacant. Multilocus sequence typing of 20 stx-positive or stx-negative E. coli O26:H11 strains showed that they were distributed into two phylogenetic groups defined by sequence type 21 (ST21) and ST29. Finally, an EspK-carrying phage was found inserted in the ssrA gene in the majority of the STEC O26:H11 strains but in only a minority of the stx-negative E. coli O26:H11 strains. The differences in the stx subtypes and Stx phage insertion sites observed in STEC O26:H11 according to their origin might reflect that strains circulating in cattle and foods are clonally distinct from those isolated from human patients. PMID:25819955

An 11-year analysis of the prevalent uropathogens and the changing pattern of Escherichia coli antibiotic resistance in 38,530 community urinary tract infections, Dublin 1999-2009.

Science.gov (United States)

Cullen, I M; Manecksha, R P; McCullagh, E; Ahmad, S; O'Kelly, F; Flynn, R; McDermott, T E D; Murphy, P; Grainger, R; Fennell, J P; Thornhill, J A

2013-03-01

Knowledge of local antimicrobial resistance patterns is essential for evidence-based empirical antibiotic prescribing, and a cutoff point of 20% has been suggested as the level of resistance at which an agent should no longer be used empirically. We sought to identify the changing incidence of causative uropathogens over an 11-year period. We also examined the trends in antibiotic resistance encountered in both the pooled urine samples and those where the causative organism was Escherichia coli. A retrospective analysis of the antimicrobial resistance within the positive community urine isolates over the 11-year period, 1999 to 2009, in a single Dublin teaching hospital was performed. In total 38,530 positive urine samples processed at our laboratory originated in the community of which 23,838 (56.7%) had E. coli as the infecting organism. The prevalence of E. coli has been increasing in recent years in community UTIs with 70.4% of UTIs in the community caused by E.coli in 2009. Ampicillin and trimethoprim were the least-active agents against E. coli with mean 11-year resistance rates of 60.8 and 31.5%, respectively. Significant trends of increasing resistance over the 11-year period were identified for trimethoprim, co-amoxyclav, cefuroxime and gentamicin. Ciprofloxacin remains a reasonable empirical antibiotic choice in this community with an 11-year resistance rate of 10.6%. Higher antibiotic resistance rates were identified in the male population and in children. Resistance rates to commonly prescribed antibiotics are increasing significantly. This data will enable evidence-based empirical prescribing which will ensure more effective treatment and lessen the emergence of resistant uropathogens in the community.

Cellular chloride and bicarbonate retention alters intracellular pH regulation in Cftr KO crypt epithelium.

Science.gov (United States)

Walker, Nancy M; Liu, Jinghua; Stein, Sydney R; Stefanski, Casey D; Strubberg, Ashlee M; Clarke, Lane L

2016-01-15

Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR), an anion channel providing a major pathway for Cl(-) and HCO3 (-) efflux across the apical membrane of the epithelium. In the intestine, CF manifests as obstructive syndromes, dysbiosis, inflammation, and an increased risk for gastrointestinal cancer. Cftr knockout (KO) mice recapitulate CF intestinal disease, including intestinal hyperproliferation. Previous studies using Cftr KO intestinal organoids (enteroids) indicate that crypt epithelium maintains an alkaline intracellular pH (pHi). We hypothesized that Cftr has a cell-autonomous role in downregulating pHi that is incompletely compensated by acid-base regulation in its absence. Here, 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein microfluorimetry of enteroids showed that Cftr KO crypt epithelium sustains an alkaline pHi and resistance to cell acidification relative to wild-type. Quantitative real-time PCR revealed that Cftr KO enteroids exhibit downregulated transcription of base (HCO3 (-))-loading proteins and upregulation of the basolateral membrane HCO3 (-)-unloader anion exchanger 2 (Ae2). Although Cftr KO crypt epithelium had increased Ae2 expression and Ae2-mediated Cl(-)/HCO3 (-) exchange with maximized gradients, it also had increased intracellular Cl(-) concentration relative to wild-type. Pharmacological reduction of intracellular Cl(-) concentration in Cftr KO crypt epithelium normalized pHi, which was largely Ae2-dependent. We conclude that Cftr KO crypt epithelium maintains an alkaline pHi as a consequence of losing both Cl(-) and HCO3 (-) efflux, which impairs pHi regulation by Ae2. Retention of Cl(-) and an alkaline pHi in crypt epithelium may alter several cellular processes in the proliferative compartment of Cftr KO intestine. Copyright © 2016 the American Physiological Society.

Cellular chloride and bicarbonate retention alters intracellular pH regulation in Cftr KO crypt epithelium

Science.gov (United States)

Walker, Nancy M.; Liu, Jinghua; Stein, Sydney R.; Stefanski, Casey D.; Strubberg, Ashlee M.

2015-01-01

Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR), an anion channel providing a major pathway for Cl− and HCO3− efflux across the apical membrane of the epithelium. In the intestine, CF manifests as obstructive syndromes, dysbiosis, inflammation, and an increased risk for gastrointestinal cancer. Cftr knockout (KO) mice recapitulate CF intestinal disease, including intestinal hyperproliferation. Previous studies using Cftr KO intestinal organoids (enteroids) indicate that crypt epithelium maintains an alkaline intracellular pH (pHi). We hypothesized that Cftr has a cell-autonomous role in downregulating pHi that is incompletely compensated by acid-base regulation in its absence. Here, 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein microfluorimetry of enteroids showed that Cftr KO crypt epithelium sustains an alkaline pHi and resistance to cell acidification relative to wild-type. Quantitative real-time PCR revealed that Cftr KO enteroids exhibit downregulated transcription of base (HCO3−)-loading proteins and upregulation of the basolateral membrane HCO3−-unloader anion exchanger 2 (Ae2). Although Cftr KO crypt epithelium had increased Ae2 expression and Ae2-mediated Cl−/HCO3− exchange with maximized gradients, it also had increased intracellular Cl− concentration relative to wild-type. Pharmacological reduction of intracellular Cl− concentration in Cftr KO crypt epithelium normalized pHi, which was largely Ae2-dependent. We conclude that Cftr KO crypt epithelium maintains an alkaline pHi as a consequence of losing both Cl− and HCO3− efflux, which impairs pHi regulation by Ae2. Retention of Cl− and an alkaline pHi in crypt epithelium may alter several cellular processes in the proliferative compartment of Cftr KO intestine. PMID:26542396

Increased adiposity, dysregulated glucose metabolism and systemic inflammation in Galectin-3 KO mice.

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Jingbo Pang

Full Text Available Obesity and type 2 diabetes are associated with increased production of Galectin-3 (Gal-3, a protein that modulates inflammation and clearance of glucose adducts. We used Lean and Diet-induced Obese (DIO WT and Gal-3 KO mice to investigate the role of Gal-3 in modulation of adiposity, glucose metabolism and inflammation. Deficiency of Gal-3 lead to age-dependent development of excess adiposity and systemic inflammation, as indicated by elevated production of acute-phase proteins, number of circulating pro-inflammatory Ly6C(high monocytes and development of neutrophilia, microcytic anemia and thrombocytosis in 20-week-old Lean and DIO male Gal-3 KO mice. This was associated with impaired fasting glucose, heightened response to a glucose tolerance test and reduced adipose tissue expression of adiponectin, Gal-12, ATGL and PPARγ, in the presence of maintained insulin sensitivity and hepatic expression of gluconeogenic enzymes in 20-week-old Gal-3 KO mice compared to their diet-matched WT controls. Expression of PGC-1α and FGF-21 in the liver of Lean Gal-3 KO mice was comparable to that observed in DIO animals. Impaired fasting glucose and altered responsiveness to a glucose load preceded development of excess adiposity and systemic inflammation, as demonstrated in 12-week-old Gal-3 KO mice. Finally, a role for the microflora in mediating the fasting hyperglycemia, but not the excessive response to a glucose load, of 12-week-old Gal-3 KO mice was demonstrated by administration of antibiotics. In conclusion, Gal-3 is an important modulator of glucose metabolism, adiposity and inflammation.

Dose-Dependent Rescue of KO Amelogenin Enamel by Transgenes in Vivo.

Science.gov (United States)

Bidlack, Felicitas B; Xia, Yan; Pugach, Megan K

2017-01-01

Mice lacking amelogenin (KO) have hypoplastic enamel. Overexpression of the most abundant amelogenin splice variant M180 and LRAP transgenes can substantially improve KO enamel, but only ~40% of the incisor thickness is recovered and the prisms are not as tightly woven as in WT enamel. This implies that the compositional complexity of the enamel matrix is required for different aspects of enamel formation, such as organizational structure and thickness. The question arises, therefore, how important the ratio of different matrix components, and in particular amelogenin splice products, is in enamel formation. Can optimal expression levels of amelogenin transgenes representing both the most abundant splice variants and cleavage product at protein levels similar to that of WT improve the enamel phenotype of KO mice? Addressing this question, our objective was here to understand dosage effects of amelogenin transgenes ( Tg ) representing the major splice variants M180 and LRAP and cleavage product CTRNC on enamel properties. Amelogenin KO mice were mated with M180 Tg , CTRNC Tg and LRAP Tg mice to generate M180 Tg and CTRNC Tg double transgene and M180 Tg , CTRNC Tg , LRAP Tg triple transgene mice with transgene hemizygosity (on one allelle) or homozygosity (on both alleles). Transgene homo- vs. hemizygosity was determined by qPCR and relative transgene expression confirmed by Western blot. Enamel volume and mineral density were analyzed by microCT, thickness and structure by SEM, and mechanical properties by Vickers microhardness testing. There were no differences in incisor enamel thickness between amelogenin KO mice with three or two different transgenes, but mice homozygous for a given transgene had significantly thinner enamel than mice hemizygous for the transgene ( p structure, but only up to a maximum of ~80% that of molar and ~40% that of incisor wild-type enamel.

Saccharomyces kudriavzevii and Saccharomyces uvarum differ from Saccharomyces cerevisiae during the production of aroma-active higher alcohols and acetate esters using their amino acidic precursors.

Science.gov (United States)

Stribny, Jiri; Gamero, Amparo; Pérez-Torrado, Roberto; Querol, Amparo

2015-07-16

Higher alcohols and acetate esters are important flavour and aroma components in the food industry. In alcoholic beverages these compounds are produced by yeast during fermentation. Although Saccharomyces cerevisiae is one of the most extensively used species, other species of the Saccharomyces genus have become common in fermentation processes. This study analyses and compares the production of higher alcohols and acetate esters from their amino acidic precursors in three Saccharomyces species: Saccharomyces kudriavzevii, Saccharomyces uvarum and S. cerevisiae. The global volatile compound analysis revealed that S. kudriavzevii produced large amounts of higher alcohols, whereas S. uvarum excelled in the production of acetate esters. Particularly from phenylalanine, S. uvarum produced the largest amounts of 2-phenylethyl acetate, while S. kudriavzevii obtained the greatest 2-phenylethanol formation from this precursor. The present data indicate differences in the amino acid metabolism and subsequent production of flavour-active higher alcohols and acetate esters among the closely related Saccharomyces species. This knowledge will prove useful for developing new enhanced processes in fragrance, flavour, and food industries. Copyright © 2015. Published by Elsevier B.V.

Ekološko oblikovana embalaža za naravno kozmetiko

OpenAIRE

Šepec, Urša

2017-01-01

Namen diplomskega dela je bil raziskati področje ekološkega oblikovanja, zahteve in priporočila pri embaliranju na področju kozmetične industrije ter načrtovati in izdelati inovativno ekološko oblikovano funkcionalno embalažo za kolekcijo trdnih mil in šamponov, ki po odprtju služi kot podstavek za milo na umivalniku. Poleg embalaže smo izdelali tudi etikete za ostale kozmetične izdelke, kot so losjoni, kreme in mazila. V teoretičnem delu je opredeljeno ekološko oblikovanje embalaže ter s...

On cycles in the transcription network of Saccharomyces cerevisiae

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Berman Piotr

2008-01-01

Full Text Available Abstract Background We investigate the cycles in the transcription network of Saccharomyces cerevisiae. Unlike a similar network of Escherichia coli, it contains many cycles. We characterize properties of these cycles and their place in the regulatory mechanism of the cell. Results Almost all cycles in the transcription network of Saccharomyces cerevisiae are contained in a single strongly connected component, which we call LSCC (L for "largest", except for a single cycle of two transcription factors. The fact that LSCC includes almost all cycles is well explained by the properties of a random graph with the same in- and out-degrees of the nodes. Among different physiological conditions, cell cycle has the most significant relationship with LSCC, as the set of 64 transcription interactions that are active in all phases of the cell cycle has overlap of 27 with the interactions of LSCC (of which there are 49. Conversely, if we remove the interactions that are active in all phases of the cell cycle (25% of interactions to transcription factors, the LSCC would have only three nodes and 5 edges, many fewer than expected. This subgraph of the transcription network consists mostly of interactions that are active only in the stress response subnetwork. We also characterize the role of LSCC in the topology of the network. We show that LSCC can be used to define a natural hierarchy in the network and that in every physiological subnetwork LSCC plays a pivotal role. Conclusion Apart from those well-defined conditions, the transcription network of Saccharomyces cerevisiae is devoid of cycles. It was observed that two conditions that were studied and that have no cycles of their own are exogenous: diauxic shift and DNA repair, while cell cycle and sporulation are endogenous. We claim that in a certain sense (slow recovery stress response is endogenous as well.

The founder of the church of Saint George at Pološko

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Pavlović Dragana

2015-01-01

Full Text Available The paper seeks to establish who was the founder of the church of Saint George at Pološko and revise the conclusions of previous researchers regarding this issue. According to the opinion commonly adopted in the scholarly literature, Jovan Dragušin, a cousin of King Dušan, had merely undertaken the construction of church at Pološko, whereas his mother was responsible for the completion of construction and painting of frescoes in the church. Through an analysis of written and visual sources the conclusion is reached that the sole person responsible for the construction and painting of the church at Pološko was the nun Maria, the mother of Jovan Dragušin. [Projekat Ministarstva nauke Republike Srbije, br. 177036: Srpska srednjovekovna umetnost i njen evropski kontekst

Altered Immune Cytokine Expression Associated with KoRV B Infection and Season in Captive Koalas

Science.gov (United States)

Higgins, Damien P.

2016-01-01

Koala (Phascolarctos cinereus) populations are increasingly vulnerable and one of the main threats is chlamydial infection. Koala retrovirus (KoRV) has been proposed as an underlying cause of the koala’s susceptibility to infection with Chlamydia and high rates of lymphoid neoplasia; however, the regionally ubiquitous, endogenous nature of this virus suggests that KoRV A infection is not sufficient for immune suppression to occur. A recently discovered exogenous variant of KoRV, KoRV B, has several structural elements that cause increased pathogenicity in related retroviruses and was associated with lymphoid neoplasia in one study. The present study assesses whether KoRV B infection is associated with alterations in immune function. Cytokine gene expression by mitogen stimulated lymphocytes of KoRV B positive (n = 5–6) and negative (n = 6–7) captive koalas was evaluated by qPCR four times (April 2014-February 2015) to control for seasonal variation. Key immune genes in the Th1 pathway (IFNγ, TNFα), Th2 pathway (IL 10, IL4, IL6) and Th17 pathway (IL17A), along with CD4:CD8 ratio, were assessed. KoRV B positive koalas showed significantly increased up-regulation of IL17A and IL10 in three out of four sampling periods and IFNγ, IL6, IL4 and TNFα in two out of four. IL17A is an immune marker for chlamydial pathogenesis in the koala; increased expression of IL17A in KoRV B positive koalas, and concurrent immune dysregulation, may explain the differences in susceptibility to chlamydial infection and severity of disease seen between individuals and populations. There was also marked seasonal variation in up-regulation for most of the cytokines and the CD4:CD8 ratio. The up-regulation in both Th1 and Th2 cytokines mirrors changes associated with immune dysregulation in humans and felids as a result of retroviral infections. This is the first report of altered immune expression in koalas infected by an exogenous variant of KoRV and also the first report of

Region-Specific Defects of Respiratory Capacities in the Ndufs4(KO Mouse Brain.

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Ernst-Bernhard Kayser

Full Text Available Lack of NDUFS4, a subunit of mitochondrial complex I (NADH:ubiquinone oxidoreductase, causes Leigh syndrome (LS, a progressive encephalomyopathy. Knocking out Ndufs4, either systemically or in brain only, elicits LS in mice. In patients as well as in KO mice distinct regions of the brain degenerate while surrounding tissue survives despite systemic complex I dysfunction. For the understanding of disease etiology and ultimately for the development of rationale treatments for LS, it appears important to uncover the mechanisms that govern focal neurodegeneration.Here we used the Ndufs4(KO mouse to investigate whether regional and temporal differences in respiratory capacity of the brain could be correlated with neurodegeneration. In the KO the respiratory capacity of synaptosomes from the degeneration prone regions olfactory bulb, brainstem and cerebellum was significantly decreased. The difference was measurable even before the onset of neurological symptoms. Furthermore, neither compensating nor exacerbating changes in glycolytic capacity of the synaptosomes were found. By contrast, the KO retained near normal levels of synaptosomal respiration in the degeneration-resistant/resilient "rest" of the brain. We also investigated non-synaptic mitochondria. The KO expectedly had diminished capacity for oxidative phosphorylation (state 3 respiration with complex I dependent substrate combinations pyruvate/malate and glutamate/malate but surprisingly had normal activity with α-ketoglutarate/malate. No correlation between oxidative phosphorylation (pyruvate/malate driven state 3 respiration and neurodegeneration was found: Notably, state 3 remained constant in the KO while in controls it tended to increase with time leading to significant differences between the genotypes in older mice in both vulnerable and resilient brain regions. Neither regional ROS damage, measured as HNE-modified protein, nor regional complex I stability, assessed by blue native

Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5.

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Blythe, Amanda; Gunasekara, Sanjika; Walshe, James; Mackay, Joel P; Hartzog, Grant A; Vrielink, Alice

2014-08-01

Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. Copyright © 2014 Elsevier Inc. All rights reserved.

Liver-specific Aquaporin 11 knockout mice show rapid vacuolization of the rough endoplasmic reticulum in periportal hepatocytes after amino acid feeding

DEFF Research Database (Denmark)

Rojek, Aleksandra; Füchtbauer, Ernst-Martin; Füchtbauer, Annette C.

2013-01-01

-specific Aqp11 KO mice, allowing us to study the role of AQP11 protein in liver of mice with normal kidney function. The unchallenged liver-specific Aqp11 KO mice have normal longevity, their livers appeared normal, and the plasma biochemistries revealed only a minor defect in lipid handling. Fasting......Aquaporin 11 (AQP11) is a protein channel expressed intracellularly in multiple organs, yet its physiological function is unclear. Aqp11 knockout (KO) mice die early due to malfunction of the kidney, a result of hydropic degeneration of proximal tubule cells. Here we report the generation of liver...... protein or larger doses of various amino acids. The fasting/refeeding challenge is associated with increased expression of markers of ER stress Grp78 and GADD153 and decreased glutathione levels, suggesting that ER stress may play role in the development of vacuoles in the AQP11-deficient hepatocytes. NMR...

Characterization of diarrhoeagenic Escherichia coli isolates in Jordanian children.

Science.gov (United States)

Shehabi, Asem A; Bulos, Najawa-Kuri; Hajjaj, Kamal G

2003-01-01

In a prospective study carried out among Jordanian children in Amman, a total of 73/250 (29.2%) stool specimens were positive for 1 or more diarrhoeagenic Escherichia coli strains using a multiplex polymerase chain reaction method. This study indicated that diarrhoeagenic E. coli isolates were found frequently more in stools of children with diarrhoea (34%) than without diarrhoea (23.1%), but without any significant difference (p > 0.05). The predominant diarrhoeagenic E. coli strains associated with diarrhoea were enteropathogenic E. coli (11.3%), followed by enterotoxigenic E. coli (9.8%) and enteroaggrative E. coli (9%), whereas in the control group these were 4.3%, 11.1% and 6%, respectively. Enteroinvasive E. coli strains (2.9%) were found only in stools of children with diarrhoea. This study revealed the absence of enterohaemorrhagic E. coli in both diarrhoeal and control stools, and found that diarrhoeagenic E. coli isolates were highly resistance to tetracycline (55%), co-trimoxazole (60%) and ampicillin (89%), which are commonly used antibiotics in Jordan.

Heat Flux Tests of the ITER FWQMs at KoHLT-1

International Nuclear Information System (INIS)

Bae, Young Dug; Kim, Suk Kwon; Shin, Hee Yun; Lee, Dong Won; Hong, Bong Guen

2009-05-01

As a party of the ITER, especially as a procurement party of the ITER blanket, we have designed the First Wall Qualification Mockup (FWQM) and fabricated five FWQMs. Two of them have been tested up to 12,690/12,020 cycles at a heat flux higher than 0.625 MW/m 2 at the KoHLT-1 facility established in the Korea Atomic Energy Research Institute (KAERI). Two KO FWQMs successfully passed the normal heat flux tests, and there was no indication of defect in the Be-to-CuCrZr joints

From Velvary, Bohemia, to the court in Vienna. The life of the imperial Kapellmeister Leopold Koželuh and a new complete edition of his keyboard sonatas

Czech Academy of Sciences Publication Activity Database

Vytlačil, Lukáš

2016-01-01

Roč. 16, č. 2 (2016), s. 7-11 ISSN 1211-0264 Institutional support: RVO:68378076 Keywords : Leopold Koželuh * composers * classicism * history of music * piano sonatas * complete editions Subject RIV: AL - Art, Architecture, Cultural Heritage

Function of brain α2B-adrenergic receptor characterized with subtype-selective α2B antagonist and KO mice.

Science.gov (United States)

Luhrs, Lauren; Manlapaz, Cynthia; Kedzie, Karen; Rao, Sandhya; Cabrera-Ghayouri, Sara; Donello, John; Gil, Daniel

2016-12-17

Noradrenergic signaling, through the α 2A and α 2C adrenergic receptors modulates the cognitive and behavioral symptoms of disorders such as schizophrenia, attention deficit hyperactivity disorder (ADHD), and addiction. However, it is unknown whether the α 2B receptor has any significant role in CNS function. The present study elucidates the potential role of the α 2B receptor in CNS function via the discovery and use of the first subtype-selective α 2B antagonist (AGN-209419), and behavioral analyses of α-receptor knockout (KO) mice. Using AGN-209419 as radioligand, α 2B receptor binding sites were identified within the olfactory bulb, cortex, thalamus, cerebellum, and striatum. Based on the observed expression patterns of α 2 subtypes in the brain, we compared α 2B KO, α 2A KO and α 2C KO mice behavioral phenotypes with their respective wild-type lines in anxiety (plus maze), compulsive (marble burying), and sensorimotor (prepulse inhibition) tasks. α 2B KO mice exhibited increased marble burying and α 2C KO mice exhibited an increased startle response to a pulse stimulus, but otherwise intact prepulse inhibition. To further explore compulsive behavior, we evaluated novelty-induced locomotor hyperactivity and found that α 2B KO and α 2C KO mice exhibited increased locomotion in the open field. Interestingly, when challenged with amphetamine, α 2C KO mice increased activity at lower doses relative to either α 2A KO or WT mice. However, α 2B KO mice exhibited stereotypy at doses of amphetamine that were only locomotor stimulatory to all other genotypes. Following co-administration of AGN-209419 with low-dose amphetamine in WT mice, stereotypy was observed, mimicking the α 2B KO phenotype. These findings suggest that the α 2B receptor is involved in CNS behaviors associated with sensorimotor gating and compulsivity, and may be therapeutically relevant for disorders such as schizophrenia, ADHD, post-traumatic stress disorder, addiction, and

Estrogen receptor-independent catechol estrogen binding activity: protein binding studies in wild-type, Estrogen receptor-alpha KO, and aromatase KO mice tissues.

Science.gov (United States)

Philips, Brian J; Ansell, Pete J; Newton, Leslie G; Harada, Nobuhiro; Honda, Shin-Ichiro; Ganjam, Venkataseshu K; Rottinghaus, George E; Welshons, Wade V; Lubahn, Dennis B

2004-06-01

Primary evidence for novel estrogen signaling pathways is based upon well-documented estrogenic responses not inhibited by estrogen receptor antagonists. In addition to 17beta-E2, the catechol estrogen 4-hydroxyestradiol (4OHE2) has been shown to elicit biological responses independent of classical estrogen receptors in estrogen receptor-alpha knockout (ERalphaKO) mice. Consequently, our research was designed to biochemically characterize the protein(s) that could be mediating the biological effects of catechol estrogens using enzymatically synthesized, radiolabeled 4-hydroxyestrone (4OHE1) and 4OHE2. Scatchard analyses identified a single class of high-affinity (K(d) approximately 1.6 nM), saturable cytosolic binding sites in several ERalphaKO estrogen-responsive tissues. Specific catechol estrogen binding was competitively inhibited by unlabeled catechol estrogens, but not by 17beta-E2 or the estrogen receptor antagonist ICI 182,780. Tissue distribution studies indicated significant binding differences both within and among various tissues in wild-type, ERalphaKO, and aromatase knockout female mice. Ligand metabolism experiments revealed extensive metabolism of labeled catechol estrogen, suggesting that catechol estrogen metabolites were responsible for the specific binding. Collectively, our data provide compelling evidence for the interaction of catechol estrogen metabolites with a novel binding protein that exhibits high affinity, specificity, and selective tissue distribution. The extensive biochemical characterization of this binding protein indicates that this protein may be a receptor, and thus may mediate ERalpha/beta-independent effects of catechol estrogens and their metabolites.

The importance of Arabidopsis glutathione peroxidase 8 for protecting Arabidopsis plant and E. coli cells against oxidative stress.

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Gaber, Ahmed

2014-01-01

Glutathione peroxidases (GPXs) are major family of the reactive oxygen species (ROS) scavenging enzymes. Recently, database analysis of the Arabidopsis genome revealed a new open-reading frame, thus increasing the total number of AtGPX gene family to eight (AtGPX1-8). The effect of plant hormones like; i. e. salicylic acid (SA), jasmonic acid (JA), abscisic acid (ABA), indoleacetic acid (IAA), and mannitol on the expression of the genes confirm that the AtGPX genes family is regulated by multiple signaling pathways. The survival rate of AtGPX8 knockout plants (KO8) was significantly decreased under heat stress compared with the wild type. Moreover, the content of malondialdehyde (MDA) and protein oxidation was significantly increased in the KO8 plant cells under heat stress. Results indicating that the deficiency of AtGPX8 accelerates the progression of oxidative stress in KO8 plants. On the other hand, the overexpression of AtGPX8 in E. coli cells enhance the growth of the recombinant enzyme on media supplemented with 0.2 mM cumene hydroperoxide, 0.3 mM H 2O 2 or 600 mM NaCl.

Studies of Saccharomyces cerevisiae and Non-Saccharomyces Yeasts during Alcoholic Fermentation

DEFF Research Database (Denmark)

Kemsawasd, Varongsiri

The early death of non-Saccharomyces yeasts during mixed culture spontaneous wine fermentation has traditionally been attributed to the lower capacity of these yeast species to withstand high levels of ethanol, low pH, and other media properties that are a part of progressing fermentation. However......, other yeast-yeast interactions, such as cell-cell contact mediated growth arrest and/or toxininduced death may also be a significant factor in the relative fragility of these non-Saccharomyces yeasts in mixed culture fermentation. In the present work we evaluate the combined roles of cell-cell contact...... and/or antimicrobial peptides on the early death of Lachancea thermotolerans during mixed culture fermentations with Saccharomyces cerevisiae. Using a specially designed double compartment fermentation system, we established that both cell-to-cell contact and antimicrobial peptides contribute...

Mixed municipal solid waste (MSW) treatment in Waste centre Spodnji Stari Grad, Krško

OpenAIRE

Kortnik, Jože; Leskovar, Jože

2015-01-01

Review paper Received: October 25, 2013 Accepted: November 7, 2013 Mixed municipal solid waste (MSW) treatment in Waste centre Spodnji Stari Grad, Krško Ravnanje z mešanimi komunalnimi odpadki v Zbirnem centru Spodnji Stari Grad, Krško Jože Kortnik1'*, Jože Leskovar2 University of Ljubljana, Faculty of Natural Sciences and Engineering, Department of Mining and Geotechnology, Aškerčeva 12, 1000 Ljubljana, Slovenia 2Kostak, d. d., Leskovška cesta 2a, 8270 Krško, Slovenia Correspo...

Spin-orbital superexchange physics emerging from interacting oxygen molecules in KO2

International Nuclear Information System (INIS)

Solovyev, I V

2008-01-01

We propose that the spin-orbital-lattice coupled phenomena, widely known for the transition-metal oxides, can be realized in molecular solids, comprising of orbitally degenerate magnetic O 2 - ions. KO 2 is one such system. Using the first-principles electronic structure calculations, we set-up an effective spin-orbital superexchange model for the low-energy molecular bands and argue that many anomalous properties of KO 2 indeed replicate the status of its orbital system in different temperature regimes

Abnormal type I collagen post-translational modification and crosslinking in a cyclophilin B KO mouse model of recessive osteogenesis imperfecta.

Science.gov (United States)

Cabral, Wayne A; Perdivara, Irina; Weis, MaryAnn; Terajima, Masahiko; Blissett, Angela R; Chang, Weizhong; Perosky, Joseph E; Makareeva, Elena N; Mertz, Edward L; Leikin, Sergey; Tomer, Kenneth B; Kozloff, Kenneth M; Eyre, David R; Yamauchi, Mitsuo; Marini, Joan C

2014-06-01

Cyclophilin B (CyPB), encoded by PPIB, is an ER-resident peptidyl-prolyl cis-trans isomerase (PPIase) that functions independently and as a component of the collagen prolyl 3-hydroxylation complex. CyPB is proposed to be the major PPIase catalyzing the rate-limiting step in collagen folding. Mutations in PPIB cause recessively inherited osteogenesis imperfecta type IX, a moderately severe to lethal bone dysplasia. To investigate the role of CyPB in collagen folding and post-translational modifications, we generated Ppib-/- mice that recapitulate the OI phenotype. Knock-out (KO) mice are small, with reduced femoral areal bone mineral density (aBMD), bone volume per total volume (BV/TV) and mechanical properties, as well as increased femoral brittleness. Ppib transcripts are absent in skin, fibroblasts, femora and calvarial osteoblasts, and CyPB is absent from KO osteoblasts and fibroblasts on western blots. Only residual (2-11%) collagen prolyl 3-hydroxylation is detectable in KO cells and tissues. Collagen folds more slowly in the absence of CyPB, supporting its rate-limiting role in folding. However, treatment of KO cells with cyclosporine A causes further delay in folding, indicating the potential existence of another collagen PPIase. We confirmed and extended the reported role of CyPB in supporting collagen lysyl hydroxylase (LH1) activity. Ppib-/- fibroblast and osteoblast collagen has normal total lysyl hydroxylation, while increased collagen diglycosylation is observed. Liquid chromatography/mass spectrometry (LC/MS) analysis of bone and osteoblast type I collagen revealed site-specific alterations of helical lysine hydroxylation, in particular, significantly reduced hydroxylation of helical crosslinking residue K87. Consequently, underhydroxylated forms of di- and trivalent crosslinks are strikingly increased in KO bone, leading to increased total crosslinks and decreased helical hydroxylysine- to lysine-derived crosslink ratios. The altered crosslink

iTRAQ-based proteome profiling of Saccharomyces cerevisiae and cryotolerant species Saccharomyces uvarum and Saccharomyces kudriavzevii during low-temperature wine fermentation.

Science.gov (United States)

García-Ríos, Estéfani; Querol, Amparo; Guillamón, José Manuel

2016-09-02

Temperature is one of the most important parameters to affect the duration and rate of alcoholic fermentation and final wine quality. Some species of the Saccharomyces genus have shown better adaptation at low temperature than Saccharomyces cerevisiae, which was the case of cryotolerant yeasts Saccharomyces uvarum and Saccharomyces kudriavzevii. In an attempt to detect inter-specific metabolic differences, we characterized the proteomic landscape of these cryotolerant species grown at 12°C and 28°C, which we compared with the proteome of S. cerevisiae (poorly adapted at low temperature). Our results showed that the main differences among the proteomic profiling of the three Saccharomyces strains grown at 12°C and 28°C lay in translation, glycolysis and amino acid metabolism. Our data corroborate previous transcriptomic results, which suggest that S. kudriavzevii is better adapted to grow at low temperature as a result of enhanced more efficient translation. Fitter amino acid biosynthetic pathways can also be mechanisms that better explain biomass yield in cryotolerant strains. Yet even at low temperature, S. cerevisiae is the most fermentative competitive species. A higher concentration of glycolytic and alcoholic fermentation enzymes in the S. cerevisiae strain might explain such greater fermentation activity. Temperature is one of the main relevant environmental variables that microorganisms have to cope with and it is also a key factor in some industrial processes that involve microorganisms. However, we are still far from understanding the molecular and physiological mechanisms of adaptation at low temperatures. The results obtained in this study provided a global atlas of the proteome changes triggered by temperature in three different species of the genus Saccharomyces with different degree of cryotolerance. These results would facilitate a better understanding of mechanisms for how yeast could adapt at the low temperature of growth. Copyright © 2016

Enzymatic activities produced by mixed Saccharomyces and non-Saccharomyces cultures: relationship with wine volatile composition.

Science.gov (United States)

Maturano, Yolanda Paola; Assof, Mariela; Fabani, María Paula; Nally, María Cristina; Jofré, Viviana; Rodríguez Assaf, Leticia Anahí; Toro, María Eugenia; Castellanos de Figueroa, Lucía Inés; Vazquez, Fabio

2015-11-01

During certain wine fermentation processes, yeasts, and mainly non-Saccharomyces strains, produce and secrete enzymes such as β-glucosidases, proteases, pectinases, xylanases and amylases. The effects of enzyme activity on the aromatic quality of wines during grape juice fermentation, using different co-inoculation strategies of non-Saccharomyces and Saccharomyces cerevisiae yeasts, were assessed in the current study. Three strains with appropriate enological performance and high enzymatic activities, BSc562 (S. cerevisiae), BDv566 (Debaryomyces vanrijiae) and BCs403 (Candida sake), were assayed in pure and mixed Saccharomyces/non-Saccharomyces cultures. β-Glucosidase, pectinase, protease, xylanase and amylase activities were quantified during fermentations. The aromatic profile of pure and mixed cultures was determined at the end of each fermentation. In mixed cultures, non-Saccharomyces species were detected until day 4-5 of the fermentation process, and highest populations were observed in MSD2 (10% S. cerevisiae/90% D. vanrijiae) and MSC1 (1% S. cerevisiae/99% C. sake). According to correlation and multivariate analysis, MSD2 presented the highest concentrations of terpenes and higher alcohols which were associated with pectinase, amylase and xylanase activities. On the other hand, MSC1 high levels of β-glucosidase, proteolytic and xylanolytic activities were correlated to esters and fatty acids. Our study contributes to a better understanding of the effect of enzymatic activities by yeasts on compound transformations that occur during wine fermentation.

Involvement of the VDE homing endonuclease and rapamycin in regulation of the Saccharomyces cerevisiae GSH11 gene encoding the high affinity glutathione transporter.

Science.gov (United States)

Miyake, Tsuyoshi; Hiraishi, Hiroyuki; Sammoto, Hiroyuki; Ono, Bun-Ichiro

2003-10-10

The Saccharomyces cerevisiae gene HGT1/GSH11 encodes the high affinity glutathione transporter and is repressed by cysteine added to the culture medium. It has been found previously that a 5'-upstream cis-element, CCGCCACAC, is responsible for regulating GSH11 expression and that several proteins bind to this element (Miyake, T., Kanayama, M., Sammoto, H., and Ono, B. (2002) Mol. Genet. Genomics 266, 1004-1011). In this report we present evidence that the most prominent of these proteins is VDE, known previously as the homing endonuclease encoded by VMA1. We show also that GSH11 is not expressed in a VDE-deleted strain and that inability to express the GSH11 of this strain is overcome by introduction of the coding region of VDE or the entire VMA1 gene. It is also found that VDE does not cut DNA in the vicinity of the GSH11 cis-element. Rapamycin, an inhibitor of the target of rapamycin (TOR) signal-transduction system, is found to enhance expression of GSH11 in a VDE-dependent manner under conditions of sulfur starvation. These results indicate that GSH11 is regulated by a system sensitive to sulfur starvation (presumably via cysteine depletion) and a more general system involving the nutritional starvation signal mediated by the TOR system. Both systems need to be operational (inhibition of TOR and sulfur starvation) for full expression of GSH11.

Explosive hazards analysis of the eutectic solution NaK and KO2

International Nuclear Information System (INIS)

Commander, J.C.

1975-06-01

Planning, preparation, conductance, and evaluation of field tests are reported to determine the explosive hazards associated with the combining of the sodium-potassium eutectic alloy (NaK) with the superoxide of potassium (KO 2 ) under various conditions of state, contamination, and detonation initiation. The planning and preparation was conducted by Aerojet Nuclear Company (ANC) at the Idaho National Engineering Laboratory, and the explosive hazards testing was done by Cook Associates, Inc., at IRECO Chemicals Pelican Point Research and Development facility in Utah. The test results showed that binary combinations of pure NaK and KO 2 could not be made to detonate, although the mixtures will spontaneously ignite and burn. However, tertiary combinations of NaK, KO 2 plus a water or hydrocarbon contaminant produced explosive hazards under a variety of conditions. The work was performed as part of the Decontamination and Decommissioning (D and D) of the first Experimental Breeder Reactor (EBR-I) and was funded by 189c I-215. (U.S.)

Pielāgojama norēķinu sistēma mākoņskaitļošanas pakalpojumiem

OpenAIRE

Surženko, Dmitrijs

2013-01-01

Šajā darbā apskatīts mākoņskaitļošanas jēdziens un mākoņskaitļošanas mākoņu veidi un to funkcionālā klasifikācija. Apskatīti norēķinu sistēmas pamati balstoties uz telekomunikāciju norēķinu sistēmas pamatiem. Aprakstīta uzņēmuma N mākoņa pārvaldības infrastruktūra. Definētas mākoņskaitļošanas brokera prasības uz uzņēmuma N piemēra. Veikts vairāku mākoņskaitļošanas pakalpojumu norēķinu sistēmas apskats un aprakstīta to atbilstība pielāgojamai norēķinu sistēmai un mākoņskaitļošanas brokera pras...

Gul'ko, descriptive, and Gruenhage compact spaces

Czech Academy of Sciences Publication Activity Database

Fabian, Marián; Montesinos, V.; Zizler, Václav

2010-01-01

Roč. 104, č. 2 (2010), s. 201-220 ISSN 1578-7303 R&D Projects: GA AV ČR IAA100190901; GA AV ČR(CZ) IAA100190610 Institutional research plan: CEZ:AV0Z10190503 Keywords : Gul'ko * descriptive * fragmentable compact spaces * network Subject RIV: BA - General Mathematics Impact factor: 0.400, year: 2010 http://link.springer.com/article/10.5052%2FRACSAM.2010.14

Effects of fermentation by Saccharomyces cerevisiae and ...

African Journals Online (AJOL)

yassine

2013-02-13

Feb 13, 2013 ... Effect of Saccharomyces cerevisiae fermentation on the ... beetroot, fermentation, Saccharomyces cerevisiae, betalain compounds. ... by Saccharomyces cerevisiae strains (González et al., .... Both red and yellow pigments were influenced during S. .... in beverages such as white wine, grape fruit, and green.

Comportamento celular e resposta antioxidante diferenciados de Saccharomyces cerevisiae e de Saccharomyces chevalieri ao metavanadato de amónio Different cellular behaviour and antioxidant response of Saccharomyces cerevisiae and Saccharomyces chevalieri growing in presence of ammonium metavanadate

Directory of Open Access Journals (Sweden)

R. Ferreira

2007-01-01

Full Text Available A fermentação do vinho é um processo microbiológico complexo que requere a presença de leveduras adaptadas a condições de stresse. No ambiente celular de organismos aeróbios ocorrem naturalmente espécies reactivas de oxigénio (ROS como subprodutos da respiração mitocondrial. A elevada reactividade destas espécies químicas pode gerar danos moleculares que, em alguns casos, levam à morte celular. Em condições fisiológicas normais ou como resposta ao stresse oxidativo, a célula pode desencadear respostas adaptativas que envolvem mecanismos antioxidantes como os enzimas glutationo redutase (GR; EC 1.6.4.2 e catalases T (CAT T; EC 1.11.1.6 e A (CAT A; EC 1.11.1.6. O vanádio, um metal pesado presente em alguns fitofármacos, pode também com portar-se como um gerador de ROS, alterando o estado redox intracelular e exercendo efeitos nocivos em leveduras expostas a quantidade excessiva deste elemento. O principal objectivo deste trabalho foi comparar o efeito do metavanadato de amónio (NH4VO3, um sal pentavalente de vanádio, na viabilidade celular e nas actividades enzimáticas GR, CAT T e CAT A das leveduras vínicas Saccharomyces cerevisiae UE-ME3 e Saccharomyces chevalieri UE-ME1. Os resultados obtidos mostram que S. chevalieri UE-ME1 revelou menor tolerância ao NH4VO3 do que S. cerevisiae UE-ME3, uma vez que culturas de S. chevalieri não sobreviveram para valores de concentração do sal de vanádio superiores a 7,5 mM enquanto que células de S. cerevisiae mantiveram-se viáveis em presença de metavanadato de amónio 75 mM. As actividades enzimáticas estudadas apresentaram em S. chevalieri valores muito inferiores aos que foram determinados em S. cerevisiae embora em ambas as espécies de levedura o NH4VO3 pareça comportarse como um indutor de stresse oxidativo ao provocar um decréscimo significativo da actividade GR (PThe fermentation of wine is a complex microbiological process which requires yeast adaptation to stress

Perbandingan Ko-kultur 2D dan 3D dengan Metode Hanging Drop untuk Menghasilkan Micro-environment yang Lebih Relevan Secara Klinis

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Radiana Dhewayani Antarianto

2017-09-01

Full Text Available Ko-kultur sel 2 dimensi (2D kurang menyerupai micro-environment seperti in vivo sedangkan ko-kultur 3 dimensi (3D membentuk mikromassa yang lebih mirip micro-environment in vivo sehingga bermanfaat dalam penelitian biologi dasar. Penelitian ini membandingkan ko-kultur 2D dan 3D sel punca serta sel stelata hepatik dengan metode hanging drop untuk menilai morfologi sel dan pembentukan sferoid dari mikromassa yang terbentuk. Studi in vitro ini dilakukan di Pusat Virologi dan Kanker Patobiologi (PRVKP UI dan laboratorium histologi FKUI pada bulan September 2015 sampai Oktober 2016 menggunakan sel punca yang diisolasi dari darah tali pusat manusia dan sel lestari LX2 (stelata hepatik manusia. Darah tali pusat disortir dengan MACS CD34 dan dianalisis flowcytometry. Ko-kultur sel punca sumsum tulang atau darah tali pusat dan LX2 dilakukan dengan metode hanging drop untuk ko-kultur 2D dan ko-kultur 3D. Triplikasi eksperimen dilakukan untuk tiap set ko-kultur. Hasilnya menunjukkan terdapat perbedaan morfologi ko-kultur 2D dan 3D hanging drop dibandingkan monokultur. Di ko-kultur 2D terdapat mikromassa dan di monokultur 2D tidak terbentuk mikromassa. Di ko-kultur 3D hanging drop, terdapat sferoid yang lebih kecil dibandingkan monokultur 3D hanging drop. Morfologi sel ko-kultur 2D dan 3D dengan metode hanging drop dibandingkan monokultur menunjukkan perubahan fenotip sel-sel yang tergabung dalam mikromassa. Normal 0 false false false false IN X-NONE X-NONE Normal 0 false false false false IN X-NONE X-NONE Molecular Basis for Saccharomyces cerevisiae Biofilm Development

DEFF Research Database (Denmark)

Andersen, Kaj Scherz

In this study, I sought to identify genes regulating the global molecular program for development of sessile multicellular communities, also known as biofilm, of the eukaryotic microorganism, Saccharomyces cerevisiae (yeast). Yeast biofilm has a clinical interest, as biofilms can cause chronic...... infections in humans. Biofilm is also interesting from an evolutionary standpoint, as an example of primitive multicellularity. By using a genome-wide screen of yeast deletion mutants, I show that 71 genes are essential for biofilm formation. Two-thirds of these genes are required for transcription of FLO11......, but only a small subset is previously described as regulators of FLO11. These results reveal that the regulation of biofilm formation and FLO11 is even more complex than what has previously been described. I find that the molecular program for biofilm formation shares many essential components with two...

Heavy metals in the air between Krompachy and Košice

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Slanèo Pavel

2000-09-01

Full Text Available The metallurgic activity of Kovohuty Krompachy (Non-ferrous Metal Works in Krompachy in the past left its visible devastation sign on the nature. It was evident that the dispersion of some elements, especially Cu, Pb, Sb and As, in the air was caused by the activity of metal works and their point sources. The properties of these metals may cause changes in health of biological objects as a result of long-term effect on the natural or communal spheres. After the treatment activities in ŽB Rudòany enterprise ended, Kovohuty Krompachy bought a treatment concentrate from ŽB Rudòany, which was freed from mercury. This concentrate still contained a lot of Cu, Sb, As, Bi and it should have been processed in Kovohuty to the so called black copper. The Municipality of the city of Košice, its department of the environment, expressed its apprehension of the possibility that this processing could cause that heavy metals may be spread in a direction of prevailing winds, i.e. south-east direction, where the city of Košice is situated. This was a reason for a task to monitor movement and occurrence of heavy metals on the route between Krompachy and Košice, as well as to monitor the share of individual sources of pollution on the quality of air in Košice urban agglomeration.Two methods applied were chosen for research purposes. The dust deposition was monitored, i.e. dust that sediments from air, its value is stated in g.m-2. 30 days-1. Another method used was the so called aspiration dust content, when air is sucked through a suitable filter and the value is stated in mg.m-3. In the given locality, there were 15 newly built or renovated sedimentation stations, where samples of sedimented dust were taken by the end of 1999 in six stages. They are marked by letters A to F. The samples of dust were analysed by atomic absorption spectrophotometry. The presented paper shows the results of phases A and D, or B and D.Aspiration method was used for fine

Anti-Trichomonas vaginalis properties of the oil of Amomum tsao-ko and its major component, geraniol.

Science.gov (United States)

Dai, Min; Peng, Cheng; Peng, Fu; Xie, Chengbin; Wang, Pinjia; Sun, Fenghui

2016-01-01

Trichomonosis, caused by the flagellate protozoan Trichomonas vaginalis, is the most common non-viral sexually transmitted disease (STD) and 5-nitroimidazole drugs are used for the treatment. However, a growing number of T. vaginalis isolates are resistant to these drugs, which make it becomes an urgent issue. The current study was designed to evaluate the anti-T. vaginalis activity of the essential oil from A. tsao-ko used in traditional Chinese medicine and as a spice and its main component, geraniol. The anti-T. vaginalis activities of A. tsao-ko essential oil and geraniol were evaluated by the minimum lethal concentration (MLC) and 50% inhibitory concentration (IC50) in vitro. The morphological changes of T. vaginalis were observed by transmission electron microscopy (TEM). Additionally, sub-MLC concentration treatment with sub-MLC A. tsao-ko essential oil and geraniol was also performed. This study shows that MLC/IC50 of A. tsao-ko essential oil was 44.97 µg/ml/22.49 µg/ml for T. vaginalis isolate Tv1, and 89.93 µg/ml/44.97 µg/ml for T. vaginalis isolate Tv2. Those of geraniol were 342.96 µg/ml/171.48 µg/ml, respectively. After A. tsao-ko essential oil or geraniol treatment, obvious similar morphological changes of T. vaginalis were observed by TEM: the nuclear membrane was damaged, nuclei were dissolved, and the chromatin was accumulated; in the cytoplasm, numerous vacuoles appeared, rough endoplasmic reticulum dilated, the number of ribosomes were reduced, organelles disintegrated, the cell membrane was partially damaged, with cytoplasmic leakage, and cell disintegration was observed. The action time did not increase the effect of A. tsao-ko essential oil or geraniol against T. vaginalis, as no significant difference was observed after sub-MLC concentration treatment for 1, 3, and 5 h with A. tsao-ko essential oil and geraniol. The study describes the first report on the activity and morphological changes of A. tsao-ko essential oil and

Magnetic properties of the alkali metal ozonides KO3, RbO3, and CsO3

International Nuclear Information System (INIS)

Lueken, H.; Deussen, M.; Jansen, M.; Hesse, W.; Schnick, W.

1987-01-01

The magnetic susceptibilities of KO 3 , RbO 3 and CsO 3 have been determined between 3.6 and 250 K. Above 50 K Curie-Weiss behaviour is observed. Magnetic moments of 1.74 μ B (KO 3 , CsO 3 ) and 1.80 μ B (RbO 3 ) calculated from the Curie-Weiss straight lines correspond with spin-only moments expected for isolated O 3 - species with one unpaired electron. The Weiss constants Θ are -34 K (KO 3 ), -23 K (RbO 3 ) and -10 K (CsO 3 ). The low temperature behaviour of KO 3 and RbO 3 (broad maxima in susceptibility at 20 and 17 K, respectively, and minima at 6 K) is typical of systems which show with decreasing temperature low-dimensional antiferromagnetic and three-dimensional magnetic ordering. Inspecting the intermolecular distances between oxygen atoms the pathways of exchange interactions are discussed. (author)

Paluch koślawy w stopie reumatycznej – leczenie operacyjne i rehabilitacja

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Agnieszka Prusinowska

2011-04-01

Full Text Available Paluch koślawy jest deformacją stopy często występującą u chorychreumatycznych (ryc. 1. W znacznym stopniu zaburza funkcjępodporową w czasie chodu oraz czynności związane z większymdynamicznym obciążeniem stopy (bieganie, skakanie. Deformacjata nasila ból, który wynika również z niedopasowania obuwia.Usprawnianie chorych z rozpoznaniem reumatoidalnego zapaleniastawów (RZS zawsze jest uzależnione od aktualnego stanu funkcjonalnegopacjenta i przeprowadzonego zabiegu operacyjnego.W artykule opisano mechanizm powstawania typowych zniekształceńprzodostopia w przebiegu RZS oraz przegląd technikoperacyjnych stosowanych w leczeniu palucha koślawego (ryc. 2.Autorzy skoncentrowali się na przedstawieniu usprawniania pokorekcji palucha koślawego z wykorzystaniem zarówno metod stosowanychw celu zmniejszenia obrzęków pooperacyjnych, jak i stabilizacjiskorygowanego chirurgicznie stawu. Do metod tych zaliczasię zarówno zabiegi z zakresu fizykoterapii, jak i ćwiczenia czynneze wsparciem kinesiotapingu (ryc. 3.

Many Saccharomyces cerevisiae Cell Wall Protein Encoding Genes Are Coregulated by Mss11, but Cellular Adhesion Phenotypes Appear Only Flo Protein Dependent.

Science.gov (United States)

Bester, Michael C; Jacobson, Dan; Bauer, Florian F

2012-01-01

The outer cell wall of the yeast Saccharomyces cerevisiae serves as the interface with the surrounding environment and directly affects cell-cell and cell-surface interactions. Many of these interactions are facilitated by specific adhesins that belong to the Flo protein family. Flo mannoproteins have been implicated in phenotypes such as flocculation, substrate adhesion, biofilm formation, and pseudohyphal growth. Genetic data strongly suggest that individual Flo proteins are responsible for many specific cellular adhesion phenotypes. However, it remains unclear whether such phenotypes are determined solely by the nature of the expressed FLO genes or rather as the result of a combination of FLO gene expression and other cell wall properties and cell wall proteins. Mss11 has been shown to be a central element of FLO1 and FLO11 gene regulation and acts together with the cAMP-PKA-dependent transcription factor Flo8. Here we use genome-wide transcription analysis to identify genes that are directly or indirectly regulated by Mss11. Interestingly, many of these genes encode cell wall mannoproteins, in particular, members of the TIR and DAN families. To examine whether these genes play a role in the adhesion properties associated with Mss11 expression, we assessed deletion mutants of these genes in wild-type and flo11Δ genetic backgrounds. This analysis shows that only FLO genes, in particular FLO1/10/11, appear to significantly impact on such phenotypes. Thus adhesion-related phenotypes are primarily dependent on the balance of FLO gene expression.

Impact of oxygenation on the performance of three non-Saccharomyces yeasts in co-fermentation with Saccharomyces cerevisiae.

Science.gov (United States)

Shekhawat, Kirti; Bauer, Florian F; Setati, Mathabatha E

2017-03-01

The sequential or co-inoculation of grape must with non-Saccharomyces yeast species and Saccharomyces cerevisiae wine yeast strains has recently become a common practice in winemaking. The procedure intends to enhance unique aroma and flavor profiles of wine. The extent of the impact of non-Saccharomyces strains depends on their ability to produce biomass and to remain metabolically active for a sufficiently long period. However, mixed-culture wine fermentations tend to become rapidly dominated by S. cerevisiae, reducing or eliminating the non-Saccharomyces yeast contribution. For an efficient application of these yeasts, it is therefore essential to understand the environmental factors that modulate the population dynamics of such ecosystems. Several environmental parameters have been shown to influence population dynamics, but their specific effect remains largely uncharacterized. In this study, the population dynamics in co-fermentations of S. cerevisiae and three non-Saccharomyces yeast species: Torulaspora delbrueckii, Lachancea thermotolerans, and Metschnikowia pulcherrima, was investigated as a function of oxygen availability. In all cases, oxygen availability strongly influenced population dynamics, but clear species-dependent differences were observed. Our data show that L. thermotolerans required the least oxygen, followed by T. delbrueckii and M. pulcherrima. Distinct species-specific chemical volatile profiles correlated in all cases with increased persistence of non-Saccharomyces yeasts, in particular increases in some higher alcohols and medium chain fatty acids. The results highlight the role of oxygen in regulating the succession of yeasts during wine fermentations and suggests that more stringent aeration strategies would be necessary to support the persistence of non-Saccharomyces yeasts in real must fermentations.

Oxygen requirement for near-UV mediated cytotoxicity of α-terthienyl to Escherichia coli and Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Arnason, T.; Chan, G.F.Q.; Wat, C.K.; Downum, K.; Towers, G.H.N.

1981-01-01

The photosensitizing activity of α-terthienyl, a naturally occurring compound from Tagetes has been studied. Fluence-response curves for the effect of α-terthienyl and near-UV radiation on Escherichia coli have been prepared. Photosensitization of E. coli was enhanced in aerobic as compared to anaerobic conditions and exogenous superoxide dismutase had a slight protective effect. Sodium azide, a quencher of 1 O 2 , protected yeast cells from photosensitization with α-terthienyl. The available evidence suggests that α-terthienyl acts as a photodynamic sensitizer in vivo. (author)

Öko tähendab edu / Liis Kängsepp

Index Scriptorium Estoniae

Kängsepp, Liis, 1981-

2008-01-01

Kaubandusturul on populaarsust kogumas mahe- ja ausalt toodetud (fair trade) kaubad. Vt. samas: Mis on mahetoit; Fair Trade; Roheline liikumine: tarbi vähem!; Mätas: pane öko peale ja müüb!; Treener Kristel Tamm jõudis ökoni oma lapsele head pakkuda tahtes

Promjene na koži lica inducirane UV-zračenjem

OpenAIRE

Bojčić, Zrinka

2016-01-01

Promjene inducirane UV zračenjem obuhvaćaju različite kliničke entitete. Zbog svoje biološke aktivnosti i sposobnosti da induciraju oštećenje DNK, UV-B zrake najvažniji su etiološki čimbenik u nastanku opeklina i karcinoma kože. Iako je UV-A zračenje glavna komponenta ultraljubičastog zračenja, manje je biološki aktivna od UV-B. UV-A zračenje ima posebno važnu ulogu u fotostarenju kože. Klinička slika akutnih promjena induciranih UV zračenjem ovisi o količini energije apsorbirane u jednoj ...

Abnormal type I collagen post-translational modification and crosslinking in a cyclophilin B KO mouse model of recessive osteogenesis imperfecta.

Directory of Open Access Journals (Sweden)

Wayne A Cabral

2014-06-01

Full Text Available Cyclophilin B (CyPB, encoded by PPIB, is an ER-resident peptidyl-prolyl cis-trans isomerase (PPIase that functions independently and as a component of the collagen prolyl 3-hydroxylation complex. CyPB is proposed to be the major PPIase catalyzing the rate-limiting step in collagen folding. Mutations in PPIB cause recessively inherited osteogenesis imperfecta type IX, a moderately severe to lethal bone dysplasia. To investigate the role of CyPB in collagen folding and post-translational modifications, we generated Ppib-/- mice that recapitulate the OI phenotype. Knock-out (KO mice are small, with reduced femoral areal bone mineral density (aBMD, bone volume per total volume (BV/TV and mechanical properties, as well as increased femoral brittleness. Ppib transcripts are absent in skin, fibroblasts, femora and calvarial osteoblasts, and CyPB is absent from KO osteoblasts and fibroblasts on western blots. Only residual (2-11% collagen prolyl 3-hydroxylation is detectable in KO cells and tissues. Collagen folds more slowly in the absence of CyPB, supporting its rate-limiting role in folding. However, treatment of KO cells with cyclosporine A causes further delay in folding, indicating the potential existence of another collagen PPIase. We confirmed and extended the reported role of CyPB in supporting collagen lysyl hydroxylase (LH1 activity. Ppib-/- fibroblast and osteoblast collagen has normal total lysyl hydroxylation, while increased collagen diglycosylation is observed. Liquid chromatography/mass spectrometry (LC/MS analysis of bone and osteoblast type I collagen revealed site-specific alterations of helical lysine hydroxylation, in particular, significantly reduced hydroxylation of helical crosslinking residue K87. Consequently, underhydroxylated forms of di- and trivalent crosslinks are strikingly increased in KO bone, leading to increased total crosslinks and decreased helical hydroxylysine- to lysine-derived crosslink ratios. The altered

Abnormal Type I Collagen Post-translational Modification and Crosslinking in a Cyclophilin B KO Mouse Model of Recessive Osteogenesis Imperfecta

Science.gov (United States)

Cabral, Wayne A.; Perdivara, Irina; Weis, MaryAnn; Terajima, Masahiko; Blissett, Angela R.; Chang, Weizhong; Perosky, Joseph E.; Makareeva, Elena N.; Mertz, Edward L.; Leikin, Sergey; Tomer, Kenneth B.; Kozloff, Kenneth M.; Eyre, David R.; Yamauchi, Mitsuo; Marini, Joan C.

2014-01-01

Cyclophilin B (CyPB), encoded by PPIB, is an ER-resident peptidyl-prolyl cis-trans isomerase (PPIase) that functions independently and as a component of the collagen prolyl 3-hydroxylation complex. CyPB is proposed to be the major PPIase catalyzing the rate-limiting step in collagen folding. Mutations in PPIB cause recessively inherited osteogenesis imperfecta type IX, a moderately severe to lethal bone dysplasia. To investigate the role of CyPB in collagen folding and post-translational modifications, we generated Ppib−/− mice that recapitulate the OI phenotype. Knock-out (KO) mice are small, with reduced femoral areal bone mineral density (aBMD), bone volume per total volume (BV/TV) and mechanical properties, as well as increased femoral brittleness. Ppib transcripts are absent in skin, fibroblasts, femora and calvarial osteoblasts, and CyPB is absent from KO osteoblasts and fibroblasts on western blots. Only residual (2–11%) collagen prolyl 3-hydroxylation is detectable in KO cells and tissues. Collagen folds more slowly in the absence of CyPB, supporting its rate-limiting role in folding. However, treatment of KO cells with cyclosporine A causes further delay in folding, indicating the potential existence of another collagen PPIase. We confirmed and extended the reported role of CyPB in supporting collagen lysyl hydroxylase (LH1) activity. Ppib−/− fibroblast and osteoblast collagen has normal total lysyl hydroxylation, while increased collagen diglycosylation is observed. Liquid chromatography/mass spectrometry (LC/MS) analysis of bone and osteoblast type I collagen revealed site-specific alterations of helical lysine hydroxylation, in particular, significantly reduced hydroxylation of helical crosslinking residue K87. Consequently, underhydroxylated forms of di- and trivalent crosslinks are strikingly increased in KO bone, leading to increased total crosslinks and decreased helical hydroxylysine- to lysine-derived crosslink ratios. The altered

Evaluation of different co-inoculation time of non-Saccharomyces/Saccharomyces yeasts in order to obtain reduced ethanol wines

Directory of Open Access Journals (Sweden)

Mestre María Victoria

2016-01-01

Full Text Available Decreasing ethanol content in wines has become one of the main objectives of winemakers in different areas of the world. The use of selected wine yeasts can be considered one of the most effective and simple tools. The aim of this study was to evaluate the effect of co-inoculation times of selected non-Saccharomyces/Saccharomyces yeasts on the reduction of ethanol levels in wines. Hanseniaspora uvarum BHu9, Starmerella bacillaris BSb55 and Candida membranaefasciens BCm71 were co-inoculate with Saccharomyces cerevisiae under fermentative conditions. Treatments assayed were: pure fermentations of S. cerevisiae BSc203 and non-Saccharomyces yeasts BHu9, BSb55 and BCm71; -co-fermentations: A-BHu9/BSc203; B-BSb55/BSc203 and C-BCm71/BSc203. These co-inoculations were carried out under mixed (simultaneous inoculation, and sequential conditions (non-Saccharomyces yeasts inoculated at initial time and S. cerevisiae at 48, 96 and 144 h. Lower fermentative efficiencies were registered when BHu9 and BSb55 remained pure more time. Conversely, the conversion efficiency was reduced in co-inocula of BCm71/BSc203, when both yeasts interact more time. Metabolites produced during all vinification processes were within acceptable concentration ranges according to the current legislations. Conclusion Time interaction during fermentation processes of non-Saccharomyces and Saccharomyces yeasts showed influence on ethanol production, and this effect would be dependent on the co-inoculated species.

Participation of SRM5/CDC28, SRM8/NET1 and SRM12/HF11 genes in activation of checkpoints of Yeast Saccharomyces Cerevisiae

International Nuclear Information System (INIS)

Kadyshevskaya, E.Yu.; Koltovaya, N.A.

2007-01-01

It is known that there are about twenty checkpoint genes in yeast Saccharomyces cerevisiae. We study participation of SRM genes selected as genes affecting genetic stability and radiosensitivity. It has been shown that srm5/cdc28-srm, srm8/net1-srm, srm12/hfil-srm mutations prevent checkpoint activation by DNA damage, particularly G0/S-checkpoint (srm5, srm8), G1/S-checkpoint (srm5, srm8, srm12), S-checkpoint (srm5, srm12) and G2-checkpoint (srm5). These data indicate, at least in budding yeast, CDC28/SRM5, HF11/ADA1/SRM12 and NET1/SRM8 genes mediate cellular response induced by DNA damage including checkpoint control

Meiotic and Mitotic Phenotypes Conferred by the blm1-1 Mutation in Saccharomyces cerevisiae and MSH4 Suppression of the Bleomycin Hypersusceptibility

Directory of Open Access Journals (Sweden)

Carol Wood Moore

2003-01-01

Full Text Available Abstract: Oxidative damage can lead to a number of diseases, and can be fatal. The blm1-1 mutation of Saccharomyces cerevisiae confers hypersusceptibility to lethal effects of the oxidative, anticancer and antifungal agent, bleomycin. For the current report, additional defects conferred by the mutation in meiosis and mitosis were investigated. The viability of spores produced during meiosis by homozygous normal BLM1/BLM1, heterozygous BLM1/blm1-1, and homozygous mutant blm1-1/blm1-1 diploid strains was studied and compared. Approximately 88% of the tetrads derived from homozygous blm1-1/blm1-1 mutant diploid cells only produced one or two viable spores. In contrast, just one tetrad among all BLM1/BLM1 and BLM1/blm1-1 tetrads only produced one or two viable spores. Rather, 94% of BLM1/BLM1 tetrads and 100% of BLM1/blm1-1 tetrads produced asci with four or three viable spores. Thus, at least one copy of the BLM1 gene is essential for the production of four viable spores after meiosis. During mitotic growth, mutant blm1-1 strains grew at reduced rates and produced cells with high frequencies of unusual morphologies compared to wild-type strains. These results indicated BLM1 is also essential for normal mitotic growth. We also investigated the suppression by the MSH4 gene, a meiosis-specific MutS homolog, of the bleomycin hypersusceptibility of blm1-1 mutant cells, and the relationship of MSH4 to BLM1. We screened a genomic library, and isolated the MSH4 gene on the basis of its ability to suppress lethal effects of bleomycin in blm1-1 cells. However, genetic mapping studies indicated that BLM1 and MSH4 are not the same gene. The possibility that chromosomal nondisjunction could be the basis for the inability of blm1-1/blm1-1 mutant cells to produce four viable spores after meiosis is discussed.

Abnormal nuclear envelopes in the striatum and motor deficits in DYT11 myoclonus-dystonia mouse models.

Science.gov (United States)

Yokoi, Fumiaki; Dang, Mai T; Zhou, Tong; Li, Yuqing

2012-02-15

DYT11 myoclonus-dystonia (M-D) is a movement disorder characterized by myoclonic jerks with dystonic symptoms and caused by mutations in paternally expressed SGCE, which codes for ε-sarcoglycan. Paternally inherited Sgce heterozygous knock-out (KO) mice exhibit motor deficits and spontaneous myoclonus. Abnormal nuclear envelopes have been reported in cellular and mouse models of early-onset DYT1 generalized torsion dystonia; however, the relationship between the abnormal nuclear envelopes and motor symptoms are not clear. Furthermore, it is not known whether abnormal nuclear envelope exists in non-DYT1 dystonia. In the present study, abnormal nuclear envelopes in the striatal medium spiny neurons (MSNs) were found in Sgce KO mice. To analyze whether the loss of ε-sarcoglycan in the striatum alone causes abnormal nuclear envelopes, motor deficits or myoclonus, we produced paternally inherited striatum-specific Sgce conditional KO (Sgce sKO) mice and analyzed their phenotypes. Sgce sKO mice exhibited motor deficits in both beam-walking and accelerated rotarod tests, while they did not exhibit abnormal nuclear envelopes, alteration in locomotion, or myoclonus. The results suggest that the loss of ε-sarcoglycan in the striatum contributes to motor deficits, while it alone does not produce abnormal nuclear envelopes or myoclonus. Development of therapies targeting the striatum to compensate for the loss of ε-sarcoglycan function may rescue the motor deficits in DYT11 M-D patients.

Saccharomyces eubayanus and Saccharomyces arboricola reside in North Island native New Zealand forests.

Science.gov (United States)

Gayevskiy, Velimir; Goddard, Matthew R

2016-04-01

Saccharomyces is one of the best-studied microbial genera, but our understanding of the global distributions and evolutionary histories of its members is relatively poor. Recent studies have altered our view of Saccharomyces' origin, but a lack of sampling from the vast majority of the world precludes a holistic perspective. We evaluate alternate Gondwanan and Far East Asian hypotheses concerning the origin of these yeasts. Being part of Gondwana, and only colonized by humans in the last ∼1000 years, New Zealand represents a unique environment for testing these ideas. Genotyping and ribosomal sequencing of samples from North Island native forest parks identified a widespread population of Saccharomyces. Whole genome sequencing identified the presence of S. arboricola and S. eubayanus in New Zealand, which is the first report of S. arboricola outside Far East Asia, and also expands S. eubayanus' known distribution to include the Oceanic region. Phylogenomic approaches place the S. arboricola population as significantly diverged from the only other sequenced Chinese isolate but indicate that S. eubayanus might be a recent migrant from South America. These data tend to support the Far East Asian origin of the Saccharomyces, but the history of this group is still far from clear. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

Implementation of KoHLT-EB DAQ System using compact RIO with EPICS

Energy Technology Data Exchange (ETDEWEB)

Chang, Dae-Sik; Kim, Suk-Kwon; Lee, Dong Won [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of); Cho, Seungyon [National Fusion Research Institute, Daejeon (Korea, Republic of)

2016-10-15

EPICS (Experimental Physics and Industrial Control System) is a collection of software tools collaboratively developed which can be integrated to provide a comprehensive and scalable control system. Currently there is an increase in use of such systems in large Physics experiments like KSTAR, ITER and DAIC (Daejeon Accelerator Ion Complex). The Korean heat load test facility (KoHLT-EB) was installed at KAERI. This facility is utilized for a qualification test of the plasma facing component (PFC) for the ITER first wall and DEMO divertor, and the thermo-hydraulic experiments. The existing data acquisition device was Agilent 34980A multifunction switch and measurement unit and controlled by Agilent VEE. In the present paper, we report the EPICS based newly upgraded KoHLT-EB DAQ system which is the advanced data acquisition system using FPGA-based reconfigurable DAQ devices like compact RIO. The operator interface of KoHLT-EB DAQ system is composed of Control-System Studio (CSS) and another server is able to archive the related data using the standalone archive tool and the archiveviewer can retrieve that data at any time in the infra-network.

NCKX3 was compensated by calcium transporting genes and bone resorption in a NCKX3 KO mouse model.

Science.gov (United States)

Yang, Hyun; Ahn, Changhwan; Shin, Eun-Kyeong; Lee, Ji-Sun; An, Beum-Soo; Jeung, Eui-Bae

2017-10-15

Gene knockout is the most powerful tool for determination of gene function or permanent modification of the phenotypic characteristics of an animal. Existing methods for gene disruption are limited by their efficiency, time required for completion and potential for confounding off-target effects. In this study, a rapid single-step approach to knockout of a targeted gene in mice using zinc-finger nucleases (ZFNs) was demonstrated for generation of mutant (knockout; KO) alleles. Specifically, ZFNs to target the sodium/calcium/potassium exchanger3 (NCKX3) gene in C57bl/6j were designed using the concept of this approach. NCKX3 KO mice were generated and the phenotypic characterization and molecular regulation of active calcium transporting genes was assessed when mice were fed different calcium diets during growth. General phenotypes such as body weight and plasma ion level showed no distinct abnormalities. Thus, the potassium/sodium/calcium exchanger of NCKX3 KO mice proceeded normally in this study. As a result, the compensatory molecular regulation of this mechanism was elucidated. Renal TRPV5 mRNA of NCKX3 KO mice increased in both male and female mice. Expression of TRPV6 mRNA was only down-regulated in the duodenum of male KO mice. Renal- and duodenal expression of PTHR and VDR were not changed; however, GR mRNA expression was increased in the kidney of NCKX3 KO mice. Depletion of the NCKX3 gene in a KO mouse model showed loss of bone mineral contents and increased plasma parathyroid hormone, suggesting that NCKX3 may play a role in regulating calcium homeostasis. Copyright © 2017 Elsevier B.V. All rights reserved.

Advanced RF-KO slow-extraction method for the reduction of spill ripple

CERN Document Server

Noda, K; Shibuya, S; Uesugi, T; Muramatsu, M; Kanazawa, M; Takada, E; Yamada, S

2002-01-01

Two advanced RF-knockout (RF-KO) slow-extraction methods have been developed at HIMAC in order to reduce the spill ripple for accurate heavy-ion cancer therapy: the dual frequency modulation (FM) method and the separated function method. As a result of simulations and experiments, it was verified that the spill ripple could be considerably reduced using these advanced methods, compared with the ordinary RF-KO method. The dual FM method and the separated function method bring about a low spill ripple within standard deviations of around 25% and of 15% during beam extraction within around 2 s, respectively, which are in good agreement with the simulation results.

Apoptosis - Triggering Effects: UVB-irradiation and Saccharomyces cerevisiae.

Science.gov (United States)

Behzadi, Payam; Behzadi, Elham

2012-12-01

The pathogenic disturbance of Saccharomyces cerevisiae is known as a rare but invasive nosocomial fungal infection. This survey is focused on the evaluation of apoptosis-triggering effects of UVB-irradiation in Saccharomyces cerevisiae. The well-growth colonies of Saccharomyces cerevisiae on Sabouraud Dextrose Agar (SDA) were irradiated within an interval of 10 minutes by UVB-light (302 nm). Subsequently, the harvested DNA molecules of control and UV-exposed yeast colonies were run through the 1% agarose gel electrophoresis comprising the luminescent dye of ethidium bromide. No unusual patterns including DNA laddering bands or smears were detected. The applied procedure for UV exposure was not effective for inducing apoptosis in Saccharomyces cerevisiae. So, it needs another UV-radiation protocol for inducing apoptosis phenomenon in Saccharomyces cerevisiae.

Saccharomyces species in the Production of Beer

Directory of Open Access Journals (Sweden)

Graham G. Stewart

2016-12-01

Full Text Available The characteristic flavour and aroma of any beer is, in large part, determined by the yeast strain employed and the wort composition. In addition, properties such as flocculation, wort fermentation ability (including the uptake of wort sugars, amino acids, and peptides, ethanol and osmotic pressure tolerance together with oxygen requirements have a critical impact on fermentation performance. Yeast management between fermentations is also a critical brewing parameter. Brewer’s yeasts are mostly part of the genus Saccharomyces. Ale yeasts belong to the species Saccharomyces cerevisiae and lager yeasts to the species Saccharomyces pastorianus. The latter is an interspecies hybrid between S. cerevisiae and Saccharomyces eubayanus. Brewer’s yeast strains are facultative anaerobes—they are able to grow in the presence or absence of oxygen and this ability supports their property as an important industrial microorganism. This article covers important aspects of Saccharomyces molecular biology, physiology, and metabolism that is involved in wort fermentation and beer production.

The ecology and evolution of non-domesticated Saccharomyces species.

Science.gov (United States)

Boynton, Primrose J; Greig, Duncan

2014-12-01

Yeast researchers need model systems for ecology and evolution, but the model yeast Saccharomyces cerevisiae is not ideal because its evolution has been affected by domestication. Instead, ecologists and evolutionary biologists are focusing on close relatives of S. cerevisiae, the seven species in the genus Saccharomyces. The best-studied Saccharomyces yeast, after S. cerevisiae, is S. paradoxus, an oak tree resident throughout the northern hemisphere. In addition, several more members of the genus Saccharomyces have recently been discovered. Some Saccharomyces species are only found in nature, while others include both wild and domesticated strains. Comparisons between domesticated and wild yeasts have pinpointed hybridization, introgression and high phenotypic diversity as signatures of domestication. But studies of wild Saccharomyces natural history, biogeography and ecology are only beginning. Much remains to be understood about wild yeasts' ecological interactions and life cycles in nature. We encourage researchers to continue to investigate Saccharomyces yeasts in nature, both to place S. cerevisiae biology into its ecological context and to develop the genus Saccharomyces as a model clade for ecology and evolution. © 2014 The Authors. Yeast published by John Wiley & Sons, Ltd.

Mākoņdatošanas izmantošana informātikas priekšmetā vidusskolā.

OpenAIRE

Jēkabsone, Kristīne

2013-01-01

Izanalizējot informātikas standartu vidusskolā izpētīta mākoņdatošanas izmantošanas iespējas vidusskolā. Veikta aptauja vidusskolas klases skolēnu vidū. Aplūkots vidusskolas izglītojamo pieejamais tehnoloģiju klāsts. Meklēta atbilde uz pētniecisko jautājumu: kādas ir mākoņdatošanas izmantošanas iespējas informātikas priekšmeta apguvē vidusskolā? Sniegti ieteikumi skolotājiem strādājot ar mākoņdatošanu. Atslēgas vārdi: informātika, mākoņdatošana, Google Drive, Skydrive, Dropbox....

Depressed nNOS expression during spine transition in the developing hippocampus of FMR1 KO mice

International Nuclear Information System (INIS)

Xu, Qin; Zhu, Zhiwei; Xu, Jialu; Gu, Weizhong; Zhao, Zhengyan

2012-01-01

Nitric oxide (NO), synthesized as needed by NO synthase (NOS), is involved in spinogenesis and synaptogenesis. Immature spine morphology is characteristic of fragile X syndrome (FXS). The objective of this research was to investigate and compare changes of postnatal neuronal NOS (nNOS) expression in the hippocampus of male fragile X mental retardation 1 gene knockout mice (FMR1 KO mice, the animal model of FXS) and male wild-type mice (WT) at postnatal day 7 (P7), P14, P21, and P28. nNOS mRNA levels were analyzed by real-time quantitative PCR (N = 4-7) and nNOS protein was estimated by Western blot (N = 3) and immunohistochemistry (N = 1). In the PCR assessment, primers 5′-GTGGCCATCGTGTCCTACCATAC-3′ and 5′-GTTTCGAGGCAGGTGGAAGCTA-3′ were used for the detection of nNOS and primers 5′-CCGTTTCTCCTGGCTCAGTTTA-3′ and 5′-CCCCAATACCACATCATCCAT-3′ were used for the detection of β-actin. Compared to the WT group, nNOS mRNA expression was significantly decreased in FMR1 KO mice at P21 (KO: 0.2857 ± 0.0150, WT: 0.5646 ± 0.0657; P < 0.05). Consistently, nNOS immunoreactivity also revealed reduced staining intensity at P21 in the FMR1 KO group. Western blot analysis validated the immunostaining results by demonstrating a significant reduction in nNOS protein levels in the FMR1 KO group compared to the WT group at P21 (KO: 0.3015 ± 0.0897, WT: 1.7542 ± 0.5455; P < 0.05). These results suggest that nNOS was involved in the postnatal development of the hippocampus in FXS and impaired NO production may retard spine maturation in FXS

Depressed nNOS expression during spine transition in the developing hippocampus of FMR1 KO mice

Energy Technology Data Exchange (ETDEWEB)

Xu, Qin; Zhu, Zhiwei; Xu, Jialu [Department of Children' s Health Care, Children' s Hospital, Zhejiang University, Hangzhou Zhejiang (China); Gu, Weizhong [Department of Pathology, Children' s Hospital, Zhejiang University, Hangzhou Zhejiang (China); Zhao, Zhengyan [Department of Children' s Health Care, Children' s Hospital, Zhejiang University, Hangzhou Zhejiang (China)

2012-10-05

Nitric oxide (NO), synthesized as needed by NO synthase (NOS), is involved in spinogenesis and synaptogenesis. Immature spine morphology is characteristic of fragile X syndrome (FXS). The objective of this research was to investigate and compare changes of postnatal neuronal NOS (nNOS) expression in the hippocampus of male fragile X mental retardation 1 gene knockout mice (FMR1 KO mice, the animal model of FXS) and male wild-type mice (WT) at postnatal day 7 (P7), P14, P21, and P28. nNOS mRNA levels were analyzed by real-time quantitative PCR (N = 4-7) and nNOS protein was estimated by Western blot (N = 3) and immunohistochemistry (N = 1). In the PCR assessment, primers 5′-GTGGCCATCGTGTCCTACCATAC-3′ and 5′-GTTTCGAGGCAGGTGGAAGCTA-3′ were used for the detection of nNOS and primers 5′-CCGTTTCTCCTGGCTCAGTTTA-3′ and 5′-CCCCAATACCACATCATCCAT-3′ were used for the detection of β-actin. Compared to the WT group, nNOS mRNA expression was significantly decreased in FMR1 KO mice at P21 (KO: 0.2857 ± 0.0150, WT: 0.5646 ± 0.0657; P < 0.05). Consistently, nNOS immunoreactivity also revealed reduced staining intensity at P21 in the FMR1 KO group. Western blot analysis validated the immunostaining results by demonstrating a significant reduction in nNOS protein levels in the FMR1 KO group compared to the WT group at P21 (KO: 0.3015 ± 0.0897, WT: 1.7542 ± 0.5455; P < 0.05). These results suggest that nNOS was involved in the postnatal development of the hippocampus in FXS and impaired NO production may retard spine maturation in FXS.

Depressed nNOS expression during spine transition in the developing hippocampus of FMR1 KO mice

Directory of Open Access Journals (Sweden)

Qin Xu

2012-12-01

Full Text Available Nitric oxide (NO, synthesized as needed by NO synthase (NOS, is involved in spinogenesis and synaptogenesis. Immature spine morphology is characteristic of fragile X syndrome (FXS. The objective of this research was to investigate and compare changes of postnatal neuronal NOS (nNOS expression in the hippocampus of male fragile X mental retardation 1 gene knockout mice (FMR1 KO mice, the animal model of FXS and male wild-type mice (WT at postnatal day 7 (P7, P14, P21, and P28. nNOS mRNA levels were analyzed by real-time quantitative PCR (N = 4-7 and nNOS protein was estimated by Western blot (N = 3 and immunohistochemistry (N = 1. In the PCR assessment, primers 5’-GTGGCCATCGTGTCCTACCATAC-3’ and 5’-GTTTCGAGGCAGGTGGAAGCTA-3’ were used for the detection of nNOS and primers 5’-CCGTTTCTCCTGGCTCAGTTTA-3’ and 5’-CCCCAATACCACATCATCCAT-3’ were used for the detection of β-actin. Compared to the WT group, nNOS mRNA expression was significantly decreased in FMR1 KO mice at P21 (KO: 0.2857 ± 0.0150, WT: 0.5646 ± 0.0657; P < 0.05. Consistently, nNOS immunoreactivity also revealed reduced staining intensity at P21 in the FMR1 KO group. Western blot analysis validated the immunostaining results by demonstrating a significant reduction in nNOS protein levels in the FMR1 KO group compared to the WT group at P21 (KO: 0.3015 ± 0.0897, WT: 1.7542 ± 0.5455; P < 0.05. These results suggest that nNOS was involved in the postnatal development of the hippocampus in FXS and impaired NO production may retard spine maturation in FXS.

Martwica kości szczęk związana ze stosowaniem bifosfonianów. Opis przypadków

Directory of Open Access Journals (Sweden)

Wojciech Leśniak

2013-06-01

Full Text Available Bifosfoniany należą do silnych inhibitorów osteoklastów. Znajdują szerokie zastosowanie w leczeniu chorób przebiegających ze wzmożoną resorpcją kości: osteoporozy, choroby Pageta i osteogenesis imperfecta. Leki te zmniejszają adhezję komórek nowotworowych do kości, ograniczają osteolizę wywołaną nowotworem i hiper‑ kalcemię. Zmniejszają dolegliwości bólowe w szpiczaku mnogim i w przerzutach do kości. Powszechnie panu‑ jące przekonanie o bezpieczeństwie spowodowało rozszerzenie wskazań do ich stosowania. Pacjenci onkolo‑ giczni przyjmujący bifosfoniany dożylnie, w wysokich dawkach są bardziej narażeni na działania niepożądane terapii niż pacjenci przyjmujący je doustnie. Jednym z najpoważniejszych powikłań jest martwica kości szczęk. W 2003 roku opisano po raz pierwszy związek pomiędzy terapią bifosfonianami a martwicą kości szczęk. W piśmiennictwie znajdujemy coraz więcej przykładów tego działania niepożądanego, także po stosowaniu leków doustnie. Mogą pojawiać się ból, obrzęk, owrzodzenie, odsłonięcie kości, przetoki, złamania patologiczne, uciążliwe dla pacjenta i obniżające jakość życia. Leczenie martwicy kości szczęk jest trudne i rzadko przynosi spodziewany efekt. Główną rolę odgrywa leczenie zachowawcze i redukcja objawów. Odstawienie bifosfonia‑ nów nie ma bezpośredniego wpływu na przebieg martwicy kości szczęk, ze względu na kumulację leku w kości. Istotne jest przygotowanie pacjenta przed rozpoczęciem podawania bifosfonianów, wymagające stałej współ‑ pracy pomiędzy lekarzami onkologami, ortopedami i chirurgami szczękowo-twarzowymi. Higiena jamy ustnej i leczenie chorób współistniejących stanowi ważny element postępowania. Celem pracy jest przedstawienie trzech przypadków martwicy kości szczęk u pacjentów poddanych leczeniu bifosfonianami.

Hexose-6-phosphate dehydrogenase contributes to skeletal muscle homeostasis independent of 11β-hydroxysteroid dehydrogenase type 1.

LENUS (Irish Health Repository)

Semjonous, Nina M

2011-01-01

Glucose-6-phosphate (G6P) metabolism by the enzyme hexose-6-phosphate dehydrogenase (H6PDH) within the sarcoplasmic reticulum lumen generates nicotinamide adenine dinucleotide phosphate (reduced) to provide the redox potential for the enzyme 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) to activate glucocorticoid (GC). H6PDH knockout (KO) mice have a switch in 11β-HSD1 activity, resulting in GC inactivation and hypothalamic-pituitary-adrenal axis activation. Importantly, H6PDHKO mice develop a type II fiber myopathy with abnormalities in glucose metabolism and activation of the unfolded protein response (UPR). GCs play important roles in muscle physiology, and therefore, we have examined the importance of 11β-HSD1 and GC metabolism in mediating aspects of the H6PDHKO myopathy. To achieve this, we examined 11β-HSD1\\/H6PDH double-KO (DKO) mice, in which 11β-HSD1 mediated GC inactivation is negated. In contrast to H6PDHKO mice, DKO mice GC metabolism and hypothalamic-pituitary-adrenal axis set point is similar to that observed in 11β-HSD1KO mice. Critically, in contrast to 11β-HSD1KO mice, DKO mice phenocopy the salient features of the H6PDHKO, displaying reduced body mass, muscle atrophy, and vacuolation of type II fiber-rich muscle, fasting hypoglycemia, increased muscle glycogen deposition, and elevated expression of UPR genes. We propose that muscle G6P metabolism through H6PDH may be as important as changes in the redox environment when considering the mechanism underlying the activation of the UPR and the ensuing myopathy in H6PDHKO and DKO mice. These data are consistent with an 11β-HSD1-independent function for H6PDH in which sarcoplasmic reticulum G6P metabolism and nicotinamide adenine dinucleotide phosphate-(oxidized)\\/nicotinamide adenine dinucleotide phosphate (reduced) redox status are important for maintaining muscle homeostasis.

Prevalence of resistance to 11 antimicrobials among Campylobacter coli isolated from pigs on 80 grower-finisher farms in Ontario

OpenAIRE

Varela, Norma P.; Friendship, Robert; Dewey, Cate

2007-01-01

We carried out a cross-sectional study to investigate antimicrobial resistance patterns of Campylobacter coli isolated from Ontario grower-finisher pigs. From January to June 2004, 1200 samples were collected from 80 farms by obtaining a constant number (15) of fecal samples per farm. Susceptibility of the isolates to 11 antimicrobial drugs was determined by the agar-dilution technique. The overall prevalence of resistance to 1 or more antimicrobials among the isolates was 99.2%. High levels ...

Lactose/whey utilization and ethanol production by transformed Saccharomyces cerevisiae cells.

Science.gov (United States)

Porro, D; Martegani, E; Ranzi, B M; Alberghina, L

1992-04-05

Strains of Saccharomyces cerevisiae transformed with a multicopy expression vector bearing both the Escherichia coli beta-galactosidase gene under the control of the upstream activating sequence of the GAL1-10 genes and the GAL4 activator gene release part of beta-galactosidase in the growth medium. This release is due to cell lysis of the older mother cells; the enzyme maintains its activity in buffered growth media. Fermentation studies with transformed yeast strains showed that the release of beta-galactosidase allowed an efficient growth on buffered media containing lactose as carbon source as well as on whey-based media. The transformed strains utilized up to 95% of the lactose and a high growth yield was obtained in rich media. High productions of ethanol were also observed in stationary phase after growth in lactose minimal media.

Rate constant and thermochemistry for K + O2 + N2 = KO2 + N2

DEFF Research Database (Denmark)

Sorvajärvi, Tapio; Viljanen, Jan; Toivonen, Juha

2015-01-01

in the form of double exponential decays of [K], which yielded both kR1 and the equilibrium constant for KO2 formation. kR1 can be summarized as 1.07 × 10-30(T/1000 K)-0.733 cm6 molecule-2 s-1. Combination with literature values leads to a recommended kR1 of 5.5 × 10-26T-1.55 exp(-10/T) cm6 molecule-2 s-1...... over 250-1320 K, with an error limit of a factor of 1.5. A vant Hoff analysis constrained to fit the computed ΔS298 yields a K-O2 bond dissociation enthalpy of 184.2 ± 4.0 kJ mol-1 at 298 K and ΔfH298(KO2) = -95.2 ± 4.1 kJ mol-1. The corresponding D0 is 181.5 ± 4.0 kJ mol-1. This value compares well...

Abnormal nuclear envelopes in the striatum and motor deficits in DYT11 myoclonus-dystonia mouse models

Science.gov (United States)

Yokoi, Fumiaki; Dang, Mai T.; Zhou, Tong; Li, Yuqing

2012-01-01

DYT11 myoclonus-dystonia (M-D) is a movement disorder characterized by myoclonic jerks with dystonic symptoms and caused by mutations in paternally expressed SGCE, which codes for ɛ-sarcoglycan. Paternally inherited Sgce heterozygous knock-out (KO) mice exhibit motor deficits and spontaneous myoclonus. Abnormal nuclear envelopes have been reported in cellular and mouse models of early-onset DYT1 generalized torsion dystonia; however, the relationship between the abnormal nuclear envelopes and motor symptoms are not clear. Furthermore, it is not known whether abnormal nuclear envelope exists in non-DYT1 dystonia. In the present study, abnormal nuclear envelopes in the striatal medium spiny neurons (MSNs) were found in Sgce KO mice. To analyze whether the loss of ɛ-sarcoglycan in the striatum alone causes abnormal nuclear envelopes, motor deficits or myoclonus, we produced paternally inherited striatum-specific Sgce conditional KO (Sgce sKO) mice and analyzed their phenotypes. Sgce sKO mice exhibited motor deficits in both beam-walking and accelerated rotarod tests, while they did not exhibit abnormal nuclear envelopes, alteration in locomotion, or myoclonus. The results suggest that the loss of ɛ-sarcoglycan in the striatum contributes to motor deficits, while it alone does not produce abnormal nuclear envelopes or myoclonus. Development of therapies targeting the striatum to compensate for the loss of ɛ-sarcoglycan function may rescue the motor deficits in DYT11 M-D patients. PMID:22080833

V labyrintech Holanovy pozdní lyriky : Nad knihou Zdeňka Kožmína

OpenAIRE

Křivánek, Vladimír

2005-01-01

This essay on the late lyric verse of Vladimír Holan, inspired by Zdeněk Kožmín’s Existencionalita, also presents an overview of previous important interpretations of Holan’s work – apart from Kožmín, it mentions the work of Přemysl Blažíček and Jiří Opelík.

Gleaning evolutionary insights from the genome sequence of a probiotic yeast Saccharomyces boulardii.

Science.gov (United States)

Khatri, Indu; Akhtar, Akil; Kaur, Kamaldeep; Tomar, Rajul; Prasad, Gandham Satyanarayana; Ramya, Thirumalai Nallan Chakravarthy; Subramanian, Srikrishna

2013-10-22

The yeast Saccharomyces boulardii is used worldwide as a probiotic to alleviate the effects of several gastrointestinal diseases and control antibiotics-associated diarrhea. While many studies report the probiotic effects of S. boulardii, no genome information for this yeast is currently available in the public domain. We report the 11.4 Mbp draft genome of this probiotic yeast. The draft genome was obtained by assembling Roche 454 FLX + shotgun data into 194 contigs with an N50 of 251 Kbp. We compare our draft genome with all other Saccharomyces cerevisiae genomes. Our analysis confirms the close similarity of S. boulardii to S. cerevisiae strains and provides a framework to understand the probiotic effects of this yeast, which exhibits unique physiological and metabolic properties.

Optimal cofactor swapping can increase the theoretical yield for chemical production in Escherichia coli and Saccharomyces cerevisiae

DEFF Research Database (Denmark)

King, Zachary A.; Feist, Adam

2014-01-01

Maintaining cofactor balance is a critical function in microorganisms, but often the native cofactor balance does not match the needs of an engineered metabolic flux state. Here, an optimization procedure is utilized to identify optimal cofactor-specificity "swaps" for oxidoreductase enzymes...... specificity of central metabolic enzymes (especially GAPD and ALCD2x) is shown to increase NADPH production and increase theoretical yields for native products in E. coli and yeast-including l-aspartate, l-lysine, l-isoleucine, l-proline, l-serine, and putrescine-and non-native products in E. coli-including 1...

The Growth Hormone Receptor Gene-Disrupted (GHR-KO) Mouse Fails to Respond to an Intermittent Fasting (IF) Diet

Science.gov (United States)

Arum, Oge; Bonkowski, Michael S.; Rocha, Juliana S.; Bartke, Andrzej

2009-01-01

SUMMARY The interaction of longevity-conferring genes with longevity-conferring diets is poorly understood. The growth hormone receptor gene-disrupted (GHR-KO) mouse is long-lived; and this longevity is not responsive to 30% caloric restriction (CR), in contrast to wild-type animals from the same strain. To determine whether this may have been limited to a particular level of dietary restriction (DR), we subjected GHR-KO mice to a different dietary restriction regimen, an intermittent fasting (IF) diet. The IF diet increased the survivorship and improved insulin sensitivity of normal males, but failed to affect either parameter in GHR-KO mice. From the results of two paradigms of dietary restriction we postulate that GHR-KO mice would be resistant to any manner of DR; potentially due to their inability to further enhance insulin sensitivity. Insulin sensitivity may be a mechanism and/or a marker of the lifespan-extending potential of an intervention. PMID:19747233

Effect of Saccharomyces, Non-Saccharomyces Yeasts and Malolactic Fermentation Strategies on Fermentation Kinetics and Flavor of Shiraz Wines

Directory of Open Access Journals (Sweden)

Heinrich du Plessis

2017-12-01

Full Text Available The use of non-Saccharomyces yeasts to improve complexity and diversify wine style is increasing; however, the interactions between non-Saccharomyces yeasts and lactic acid bacteria (LAB have not received much attention. This study investigated the interactions of seven non-Saccharomyces yeast strains of the genera Candida, Hanseniaspora, Lachancea, Metschnikowia and Torulaspora in combination with S. cerevisiae and three malolactic fermentation (MLF strategies in a Shiraz winemaking trial. Standard oenological parameters, volatile composition and sensory profiles of wines were investigated. Wines produced with non-Saccharomyces yeasts had lower alcohol and glycerol levels than wines produced with S. cerevisiae only. Malolactic fermentation also completed faster in these wines. Wines produced with non-Saccharomyces yeasts differed chemically and sensorially from wines produced with S. cerevisiae only. The Candida zemplinina and the one L. thermotolerans isolate slightly inhibited LAB growth in wines that underwent simultaneous MLF. Malolactic fermentation strategy had a greater impact on sensory profiles than yeast treatment. Both yeast selection and MLF strategy had a significant effect on berry aroma, but MLF strategy also had a significant effect on acid balance and astringency of wines. Winemakers should apply the optimal yeast combination and MLF strategy to ensure fast completion of MLF and improve wine complexity.

Farklı stres koşullarında Rhodotorula glutinis’ten antioksidan üretimi için biyoproses koşullarının geliştirilmesi

OpenAIRE

ÜNLÜ BÜYÜKTOPÇU, Ayşe Ezgi

2012-01-01

Source of spill ripple in the RF-KO slow-extraction method with FM and AM

International Nuclear Information System (INIS)

Noda, K.; Furukawa, T.; Shibuya, S.; Muramatsu, M.; Uesugi, T.; Kanazawa, M.; Torikoshi, M.; Takada, E.; Yamada, S.

2002-01-01

The RF-knockout (RF-KO) slow-extraction method with frequency modulation (FM) and amplitude modulation (AM) has brought high-accuracy irradiation to the treatment of a cancer tumor moving with respiration, because of a quick response to beam start/stop. However, a beam spill extracted from a synchrotron ring through RF-KO slow-extraction has a huge ripple with a frequency of around 1 kHz related to the FM. The spill ripple will disturb the lateral dose distribution in the beam scanning methods. Thus, the source of the spill ripple has been investigated through experiments and simulations. There are two tune regions for the extraction process through the RF-KO method: the extraction region and the diffusion region. The particles in the extraction region can be extracted due to amplitude growth through the transverse RF field, only when its frequency matches with the tune in the extraction region. For a large chromaticity, however, the particles in the extraction region can be extracted through the synchrotron oscillation, even when the frequency does not match with the tune in the extraction region. Thus, the spill structure during one period of the FM strongly depends on the horizontal chromaticity. They are repeated with the repetition frequency of the FM, which is the very source of the spill ripple in the RF-KO method

Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Bojsen, Rasmus K; Andersen, Kaj Scherz; Regenberg, Birgitte

2012-01-01

Microbial biofilms can be defined as multi-cellular aggregates adhering to a surface and embedded in an extracellular matrix (ECM). The nonpathogenic yeast, Saccharomyces cerevisiae, follows the common traits of microbial biofilms with cell-cell and cell-surface adhesion. S. cerevisiae is shown t...

Jeast (Saccharomyces cerevisial) mutants with enhanced induced mutagenesis

International Nuclear Information System (INIS)

Ivanov, E.L.; Koval'tsova, S.V.; Korolev, V.G.

1987-01-01

The influence of him1-1, him2-1, him3-1 and himX mutations on induction frequency and specificity of UV-induced adenine-dependent mutations in the yeast Saccharomyces cerevisiae has been. Him mutations do not render haploid cells more sensitive to the lethal action of UV-light; however, in him strains adeine-dependent mutations (ade, ade2) were induced more frequently (1.5-2-fold), as compared to the HIM strain. An analysis of the molecular nature of ade2 mutants revealed than him1-1, him2-1, and himX mutations increase specifically the yield of transitions (AT-GC and GC→AT), whereas in the him3-1, strain the yield of transversions was enhanced as well. We suggest him mutations analysed to affect specific repair pathway for mismatch correction

Scyntygrafia kości w diagnostyce reumatoidalnego zapalenia stawów

Directory of Open Access Journals (Sweden)

Witold Tłustochowicz

2010-04-01

Full Text Available Celem pracy była ocena przydatności trójfazowej dynamicznejscyntygrafii kości w diagnostyce reumatoidalnego zapalenia stawów.Badaniem objęto 39 chorych hospitalizowanych i diagnozowanychz powodu dolegliwości stawowych. Przeprowadzono diagnostykęobejmującą: badanie podmiotowe, przedmiotowe, badania laboratoryjneoraz obrazowe, w tym trójfazową scyntygrafię kości z użyciemtechnetu 99m. Scyntygraficzne cechy zapalenia stawówstwierdzano, gdy obserwowano wzmożone gromadzenie znacznikawe wszystkich trzech fazach badania.Po ukończeniu diagnostyki rozpoznano u 13 chorych wczesne reumatoidalnezapalenie stawów, u 4 reumatoidalne zapalenie stawów(łącznie 17 pacjentów, a u 1 niezróżnicowane zapalenie stawów.Scyntygrafia kości wykazała cechy zapalenia stawów u 13spośród nich (72,2%, u 4 osób (22,2% wzmożone gromadzeniestwierdzono tylko w fazie statycznej, a u 1 pacjenta (5,6% uzyskanoprawidłowy wynik badania. U 21 chorych wykluczono chorobęzapalną stawów (u 19 rozpoznano fibromialgię, u 2 chorobęzwyrodnieniową. W grupie bez zapalenia stawów u 14 pacjentów(66,6% nie stwierdzono istotnych nieprawidłowości w obraziescyntygraficznym, u 6 (28,6% występowało wzmożone gromadzenieznacznika w badaniu statycznym, a u 1 chorego (4,8% opisanoscyntygraficzne cechy zapalenia stawów.Wbadanej grupie czułość scyntygrafii dynamicznej kości w wykrywaniuzapalenia stawów wynosiła 72,2% (95% CI: 57,5–76,8, a swoistość 95,2% (95% CI: 82,8–99,1. Dodatni wynik badaniadynamicznego wskazuje z dużym prawdopodobieństwem naobecność zapalenia stawów [PPV 92,9% (95% CI: 74,2–98,7].Mniej wiarygodny był ujemny wynik badania [NPV 80% (95% CI:69,5–83,3]. Wyniki niniejszej pracy wskazują, że trójfazowa scyntygrafia dynamicznakości może mieć zastosowanie w diagnostyce reumatoidalnegozapalenia stawów.

A Note on Ko-Type O-Ending Syllables in Old Japanese

Science.gov (United States)

Unger, J. Marshall

1975-01-01

Further evidence is offered to support the argument that ko-type o-ending syllables may have been secondary in nature in Old Japanese. Pre-Old Japanese indicates the */uwa/ sequence was manifested as /uwe/ and /uwu/ endings of o-ending syllable verbs. (SCC)

Source of spill ripple in the RF-KO slow-extraction method with FM and AM

CERN Document Server

Noda, K; Shibuya, S; Muramatsu, M; Uesugi, T; Kanazawa, M; Torikoshi, M; Takada, E; Yamada, S

2002-01-01

The RF-knockout (RF-KO) slow-extraction method with frequency modulation (FM) and amplitude modulation (AM) has brought high-accuracy irradiation to the treatment of a cancer tumor moving with respiration, because of a quick response to beam start/stop. However, a beam spill extracted from a synchrotron ring through RF-KO slow-extraction has a huge ripple with a frequency of around 1 kHz related to the FM. The spill ripple will disturb the lateral dose distribution in the beam scanning methods. Thus, the source of the spill ripple has been investigated through experiments and simulations. There are two tune regions for the extraction process through the RF-KO method: the extraction region and the diffusion region. The particles in the extraction region can be extracted due to amplitude growth through the transverse RF field, only when its frequency matches with the tune in the extraction region. For a large chromaticity, however, the particles in the extraction region can be extracted through the synchrotron ...

Key on demand (KoD) for software-defined optical networks secured by quantum key distribution (QKD).

Science.gov (United States)

Cao, Yuan; Zhao, Yongli; Colman-Meixner, Carlos; Yu, Xiaosong; Zhang, Jie

2017-10-30

Software-defined optical networking (SDON) will become the next generation optical network architecture. However, the optical layer and control layer of SDON are vulnerable to cyberattacks. While, data encryption is an effective method to minimize the negative effects of cyberattacks, secure key interchange is its major challenge which can be addressed by the quantum key distribution (QKD) technique. Hence, in this paper we discuss the integration of QKD with WDM optical networks to secure the SDON architecture by introducing a novel key on demand (KoD) scheme which is enabled by a novel routing, wavelength and key assignment (RWKA) algorithm. The QKD over SDON with KoD model follows two steps to provide security: i) quantum key pools (QKPs) construction for securing the control channels (CChs) and data channels (DChs); ii) the KoD scheme uses RWKA algorithm to allocate and update secret keys for different security requirements. To test our model, we define a security probability index which measures the security gain in CChs and DChs. Simulation results indicate that the security performance of CChs and DChs can be enhanced by provisioning sufficient secret keys in QKPs and performing key-updating considering potential cyberattacks. Also, KoD is beneficial to achieve a positive balance between security requirements and key resource usage.

Molecular epidemiology over an 11-year period (2000 to 2010) of extended-spectrum β-lactamase-producing Escherichia coli causing bacteremia in a centralized Canadian region

NARCIS (Netherlands)

G. Peirano (G.); A.K. van der Bij (Akke); S. Gregson (Simon); J.D.D. Pitout (J. D D)

2012-01-01

textabstractA study was designed to assess the importance of sequence types among extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolates causing bacteremia over an 11-year period (2000 to 2010) in a centralized Canadian region. A total of 197 patients with incident infections were

„Nekad su mi znali reći da nisam bolja od svog sina“ - stigma obitelji temeljem intelektualnih teškoća člana obitelji

OpenAIRE

Buljevac, Marko; Leutar, Zdravka

2017-01-01

Rad se bavi stigmom obitelji temeljem intelektualnih teškoća člana obitelji. Cilj ovog istraživanja bio je dobiti uvid u perspektive članova uže obitelji osoba s različitim stupnjem intelektualnih teškoća o stigmi obitelji temeljem intelektualnih teškoća. Istraživačko pitanje bilo je: „Kako članovi obitelji osoba s intelektualnim teškoćama doživljavaju stigmu obitelji?“. Sudionici istraživanja bile se 22 osobe koje su članovi uže obitelji osoba s različitim stupnjem intelektualnih teškoća (10...

Photodynamic DNA damage induced by phycocyanin and its repair in Saccharomyces cerevisiae

Directory of Open Access Journals (Sweden)

M. Pádula

1999-09-01

Full Text Available In the present study, we analyzed DNA damage induced by phycocyanin (PHY in the presence of visible light (VL using a set of repair endonucleases purified from Escherichia coli. We demonstrated that the profile of DNA damage induced by PHY is clearly different from that induced by molecules that exert deleterious effects on DNA involving solely singlet oxygen as reactive species. Most of PHY-induced lesions are single strand breaks and, to a lesser extent, base oxidized sites, which are recognized by Nth, Nfo and Fpg enzymes. High pressure liquid chromatography coupled to electrochemical detection revealed that PHY photosensitization did not induce 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo at detectable levels. DNA repair after PHY photosensitization was also investigated. Plasmid DNA damaged by PHY photosensitization was used to transform a series of Saccharomyces cerevisiae DNA repair mutants. The results revealed that plasmid survival was greatly reduced in rad14 mutants, while the ogg1 mutation did not modify the plasmid survival when compared to that in the wild type. Furthermore, plasmid survival in the ogg1 rad14 double mutant was not different from that in the rad14 single mutant. The results reported here indicate that lethal lesions induced by PHY plus VL are repaired differently by prokaryotic and eukaryotic cells. Morever, nucleotide excision repair seems to play a major role in the recognition and repair of these lesions in Saccharomyces cerevisiae.

Effects of fermentation by Saccharomyces cerevisiae and ...

African Journals Online (AJOL)

yassine

2013-02-13

Feb 13, 2013 ... Full Length Research Paper. Effect of Saccharomyces cerevisiae fermentation on the ... 2003). Besides, several alcoholic beverages such as wine or liqueurs are obtained from fruit juices fermented by Saccharomyces ..... (2003). Kinetics of pigment release from hairy root cultures of Beta vulgaris under the ...

Experience with Saccharomyces boulardii Probiotic in Oncohaematological Patients.

Science.gov (United States)

Sulik-Tyszka, Beata; Snarski, Emilian; Niedźwiedzka, Magda; Augustyniak, Małgorzata; Myhre, Thorvald Nilsen; Kacprzyk, Anna; Swoboda-Kopeć, Ewa; Roszkowska, Marta; Dwilewicz-Trojaczek, Jadwiga; Jędrzejczak, Wiesław Wiktor; Wróblewska, Marta

2018-06-01

Very few reports have been published to date on the bloodstream infections caused by Saccharomyces spp. in oncohaematological patients, and there are no guidelines on the use of this probiotic microorganism in this population. We describe the use of probiotic preparation containing Saccharomyces boulardii in a large group of oncohaematological patients. We retrospectively analysed the data from 32,000 patient hospitalisations at the haematological centre during 2011-2013 (including 196 haematopoietic stem cell transplant recipients) in a tertiary care university-affiliated hospital. During the study period, 2270 doses of Saccharomyces boulardii probiotic were administered to the oncohaematological patients. In total, 2816 mycological cultures were performed, out of which 772 (27.4%) were positive, with 52 indicating digestive tract colonisation by Saccharomyces spp., mainly in patients with acute myeloid leukaemia (AML), myelodysplastic syndrome (MDS) or multiple myeloma (MM). While colonised, they were hospitalised for 1683 days and 416 microbiological cultures of their clinical samples were performed. In the studied group of patients, there were six blood cultures positive for fungi; however, they comprised Candida species: two C. glabrata, one C. albicans, one C. krusei, one C. tropicalis and one C. parapsilosis. There was no blood culture positive for Saccharomyces spp. Our study indicates that despite colonisation of many oncohaematological patients with Saccharomyces spp., there were no cases of fungal sepsis caused by this species.

Saccharomyces eubayanus and Saccharomyces uvarum associated with the fermentation of Araucaria araucana seeds in Patagonia.

Science.gov (United States)

Rodríguez, M Eugenia; Pérez-Través, Laura; Sangorrín, Marcela P; Barrio, Eladio; Lopes, Christian A

2014-09-01

Mudai is a traditional fermented beverage, made from the seeds of the Araucaria araucana tree by Mapuche communities. The main goal of the present study was to identify and characterize the yeast microbiota responsible of Mudai fermentation as well as from A. araucana seeds and bark from different locations in Northern Patagonia. Only Hanseniaspora uvarum and a commercial bakery strain of Saccharomyces cerevisiae were isolated from Mudai and all Saccharomyces isolates recovered from A. araucana seed and bark samples belonged to the cryotolerant species Saccharomyces eubayanus and Saccharomyces uvarum. These two species were already reported in Nothofagus trees from Patagonia; however, this is the first time that they were isolated from A. araucana, which extends their ecological distribution. The presence of these species in A. araucana seeds and bark samples, led us to postulate a potential role for them as the original yeasts responsible for the elaboration of Mudai before the introduction of commercial S. cerevisiae cultures. The molecular and genetic characterization of the S. uvarum and S. eubayanus isolates and their comparison with European S. uvarum strains and S. eubayanus hybrids (S. bayanus and S. pastorianus), allowed their ecology and evolution us to be examined. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Comparative ultraviolet action spectra (254-320 nm) of five wild-type eukaryotic microorganisms and Escherichia coli

International Nuclear Information System (INIS)

Calkins, J.; Wheeler, J.S.; Keller, C.I.; Colley, E.; Hazle, J.D.

1988-01-01

The action spectra of five eukaryotic organisms and the prokaryote, Escherichia coli, were examined over the wavelength range, 254-320 nm. Both the repair competent and three repair defective strains (E. coli, Caenorhabditis elegans, Saccharomyces) were examined. Tetrahymena pyriformis action spectra were performed with and without the excision repair inhibitor caffeine present. Others have observed that lethality, mutation, and the production of pyrimidine dimers show much the same wavelength dependence as DNA absorption. The results presented here demonstrate several action spectra which deviate from the DNA absorption spectra. Ultraviolet sensitization ratios (repair competent/repair defective) were also examined and were shown to change over the wavelength range. These findings suggest that DNA may not be the only important chromophore leading to cell death in the uv wavelength range studied. Since uv-B is of major importance in solar uv damage, these findings may also yield important implications for solar uv studies

Outlining a future for non-Saccharomyces yeasts: selection of putative spoilage wine strains to be used in association with Saccharomyces cerevisiae for grape juice fermentation.

Science.gov (United States)

Domizio, Paola; Romani, Cristina; Lencioni, Livio; Comitini, Francesca; Gobbi, Mirko; Mannazzu, Ilaria; Ciani, Maurizio

2011-06-30

The use of non-Saccharomyces yeasts that are generally considered as spoilage yeasts, in association with Saccharomyces cerevisiae for grape must fermentation was here evaluated. Analysis of the main oenological characteristics of pure cultures of 55 yeasts belonging to the genera Hanseniaspora, Pichia, Saccharomycodes and Zygosaccharomyces revealed wide biodiversity within each genus. Moreover, many of these non-Saccharomyces strains had interesting oenological properties in terms of fermentation purity, and ethanol and secondary metabolite production. The use of four non-Saccharomyces yeasts (one per genus) in mixed cultures with a commercial S. cerevisiae strain at different S. cerevisiae/non-Saccharomyces inoculum ratios was investigated. This revealed that most of the compounds normally produced at high concentrations by pure cultures of non-Saccharomyces, and which are considered detrimental to wine quality, do not reach threshold taste levels in these mixed fermentations. On the other hand, the analytical profiles of the wines produced by these mixed cultures indicated that depending on the yeast species and the S. cerevisiae/non-Saccharomyces inoculum ratio, these non-Saccharomyces yeasts can be used to increase production of polysaccharides and to modulate the final concentrations of acetic acid and volatile compounds, such as ethyl acetate, phenyl-ethyl acetate, 2-phenyl ethanol, and 2-methyl 1-butanol. Copyright © 2011 Elsevier B.V. All rights reserved.

Cluster and principal component analysis based on SSR markers of Amomum tsao-ko in Jinping County of Yunnan Province

Science.gov (United States)

Ma, Mengli; Lei, En; Meng, Hengling; Wang, Tiantao; Xie, Linyan; Shen, Dong; Xianwang, Zhou; Lu, Bingyue

2017-08-01

Amomum tsao-ko is a commercial plant that used for various purposes in medicinal and food industries. For the present investigation, 44 germplasm samples were collected from Jinping County of Yunnan Province. Clusters analysis and 2-dimensional principal component analysis (PCA) was used to represent the genetic relations among Amomum tsao-ko by using simple sequence repeat (SSR) markers. Clustering analysis clearly distinguished the samples groups. Two major clusters were formed; first (Cluster I) consisted of 34 individuals, the second (Cluster II) consisted of 10 individuals, Cluster I as the main group contained multiple sub-clusters. PCA also showed 2 groups: PCA Group 1 included 29 individuals, PCA Group 2 included 12 individuals, consistent with the results of cluster analysis. The purpose of the present investigation was to provide information on genetic relationship of Amomum tsao-ko germplasm resources in main producing areas, also provide a theoretical basis for the protection and utilization of Amomum tsao-ko resources.

Lycopene overproduction in Saccharomyces cerevisiae through combining pathway engineering with host engineering.

Science.gov (United States)

Chen, Yan; Xiao, Wenhai; Wang, Ying; Liu, Hong; Li, Xia; Yuan, Yingjin

2016-06-21

Microbial production of lycopene, a commercially and medically important compound, has received increasing concern in recent years. Saccharomyces cerevisiae is regarded as a safer host for lycopene production than Escherichia coli. However, to date, the lycopene yield (mg/g DCW) in S. cerevisiae was lower than that in E. coli and did not facilitate downstream extraction process, which might be attributed to the incompatibility between host cell and heterologous pathway. Therefore, to achieve lycopene overproduction in S. cerevisiae, both host cell and heterologous pathway should be delicately engineered. In this study, lycopene biosynthesis pathway was constructed by integration of CrtE, CrtB and CrtI in S. cerevisiae CEN.PK2. When YPL062W, a distant genetic locus, was deleted, little acetate was accumulated and approximately 100 % increase in cytosolic acetyl-CoA pool was achieved relative to that in parental strain. Through screening CrtE, CrtB and CrtI from diverse species, an optimal carotenogenic enzyme combination was obtained, and CrtI from Blakeslea trispora (BtCrtI) was found to have excellent performance on lycopene production as well as lycopene proportion in carotenoid. Then, the expression level of BtCrtI was fine-tuned and the effect of cell mating types was also evaluated. Finally, potential distant genetic targets (YJL064W, ROX1, and DOS2) were deleted and a stress-responsive transcription factor INO2 was also up-regulated. Through the above modifications between host cell and carotenogenic pathway, lycopene yield was increased by approximately 22-fold (from 2.43 to 54.63 mg/g DCW). Eventually, in fed-batch fermentation, lycopene production reached 55.56 mg/g DCW, which is the highest reported yield in yeasts. Saccharomyces cerevisiae was engineered to produce lycopene in this study. Through combining host engineering (distant genetic loci and cell mating types) with pathway engineering (enzyme screening and gene fine-tuning), lycopene yield was

Genomic insights into the Saccharomyces sensu stricto complex.

Science.gov (United States)

Borneman, Anthony R; Pretorius, Isak S

2015-02-01

The Saccharomyces sensu stricto group encompasses species ranging from the industrially ubiquitous yeast Saccharomyces cerevisiae to those that are confined to geographically limited environmental niches. The wealth of genomic data that are now available for the Saccharomyces genus is providing unprecedented insights into the genomic processes that can drive speciation and evolution, both in the natural environment and in response to human-driven selective forces during the historical "domestication" of these yeasts for baking, brewing, and winemaking. Copyright © 2015 by the Genetics Society of America.

Saccharomyces cerevisiae var. boulardii fungemia following probiotic treatment

Directory of Open Access Journals (Sweden)

Marcelo C. Appel-da-Silva

2017-12-01

Full Text Available Probiotics are commonly prescribed as an adjuvant in the treatment of antibiotic-associated diarrhea caused by Clostridium difficile. We report the case of an immunocompromised 73-year-old patient on chemotherapy who developed Saccharomyces cerevisiae var. boulardii fungemia in a central venous catheter during treatment of antibiotic-associated pseudomembranous colitis with the probiotic Saccharomyces cerevisiae var. boulardii. Fungemia was resolved after interruption of probiotic administration without the need to replace the central venous line. Keywords: Saccharomyces, Probiotics, Fungemia, Critical illness, Clostridium difficile

The Saccharomyces cerevisiae RAD30 gene, a homologue of Escherichia coli dinB and umuC, is DNA damage inducible and functions in a novel error-free postreplication repair mechanism

Energy Technology Data Exchange (ETDEWEB)

McDonald, J. P. [NIH, Bethesda, MD. (United States); Levine, A. S.; Woodgate, R.

1997-12-15

Damage-inducible mutagenesis in prokaryotes is largely dependent upon the activity of the UmuD'C-like proteins. Since many DNA repair processes are structurally and/or functionally conserved between prokaryotes and eukaryotes, we investigated the role of RAD30, a previously uncharacterized Saccharomyces cerevisiae DNA repair gene related to the Escherichia coli dinB, umuC and S. cerevisiae REV1 genes, in UV resistance and UV-induced mutagenesis. Similar to its prokaryotic homologues, RAD30 was found to be damage inducible. Like many S. cerevisiae genes involved in error-prone DNA repair, epistasis analysis clearly places RAD30 in the RAD6 group and rad30 mutants display moderate UV sensitivity reminiscent of rev mutants. However, unlike rev mutants, no defect in UV-induced reversion was seen in rad30 strains. While rad6 and rad18 are both epistatic to rad30, no epistasis was observed with rev1, rev3, rev7 or rad5, all of which are members of the RAD6 epistasis group. These findings suggest that RD30 participates in a novel error-free repair pathway dependent on RAD6 and RAD18, but independent of REV1, REV3, REV7 and RAD5. (author)

Experimental transmission of AA amyloidosis by injecting the AA amyloid protein into interleukin-1 receptor antagonist knockout (IL-1raKO) mice.

Science.gov (United States)

Watanabe, K; Uchida, K; Chambers, J K; Tei, M; Shoji, A; Ushio, N; Nakayama, H

2015-05-01

The incidence of AA amyloidosis is high in humans with rheumatoid arthritis and several animal species, including cats and cattle with prolonged inflammation. AA amyloidosis can be experimentally induced in mice using severe inflammatory stimuli and a coinjection of AA amyloid; however, difficulties have been associated with transmitting AA amyloidosis to a different animal species, and this has been attributed to the "species barrier." The interleukin-1 receptor antagonist knockout (IL-1raKO) mouse, a rodent model of human rheumatoid arthritis, has been used in the transmission of AA amyloid. When IL-1raKO and BALB/c mice were intraperitoneally injected with mouse AA amyloid together with a subcutaneous pretreatment of 2% AgNO3, all mice from both strains that were injected with crude or purified murine AA amyloid developed AA amyloidosis. However, the amyloid index, which was determined by the intensity of AA amyloid deposition, was significantly higher in IL-1raKO mice than in BALB/c mice. When IL-1raKO and BALB/c mice were injected with crude or purified bovine AA amyloid together with the pretreatment, 83% (5/6 cases) and 38% (3/8 cases) of IL-1raKO mice and 17% (1/6 cases) and 0% (0/6 cases) of BALB/c mice, respectively, developed AA amyloidosis. Similarly, when IL-1raKO and BALB/c mice were injected with crude or purified feline AA amyloid, 33% (2/6 cases) and 88% (7/8 cases) of IL-1raKO mice and 0% (0/6 cases) and 29% (2/6 cases) of BALB/c mice, respectively, developed AA amyloidosis. These results indicated that IL-1raKO mice are a useful animal model for investigating AA amyloidogenesis. © The Author(s) 2014.

Glucose repression in Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Kayikci, Omur; Nielsen, Jens

2015-01-01

Glucose is the primary source of energy for the budding yeast Saccharomyces cerevisiae. Although yeast cells can utilize a wide range of carbon sources, presence of glucose suppresses molecular activities involved in the use of alternate carbon sources as well as it represses respiration and gluc......Glucose is the primary source of energy for the budding yeast Saccharomyces cerevisiae. Although yeast cells can utilize a wide range of carbon sources, presence of glucose suppresses molecular activities involved in the use of alternate carbon sources as well as it represses respiration...

Liver steatosis study_PFAA treated Wild type and PPAR KO mouse data

Data.gov (United States)

U.S. Environmental Protection Agency — Data set 1 consists of the experimental data for the Wild Type and PPAR KO animal study and includes data used to prepare Figures 1-4 and Table 1 of the Das et al,...

Partial amino acid sequence of the branched chain amino acid aminotransferase (TmB) of E. coli JA199 pDU11

International Nuclear Information System (INIS)

Feild, M.J.; Armstrong, F.B.

1987-01-01

E. coli JA199 pDU11 harbors a multicopy plasmid containing the ilv GEDAY gene cluster of S. typhimurium. TmB, gene product of ilv E, was purified, crystallized, and subjected to Edman degradation using a gas phase sequencer. The intact protein yielded an amino terminal 31 residue sequence. Both carboxymethylated apoenzyme and [ 3 H]-NaBH-reduced holoenzyme were then subjected to digestion by trypsin. The digests were fractionated using reversed phase HPLC, and the peptides isolated were sequenced. The borohydride-treated holoenzyme was used to isolate the cofactor-binding peptide. The peptide is 27 residues long and a comparison with known sequences of other aminotransferases revealed limited homology. Peptides accounting for 211 of 288 predicted residues have been sequenced, including 9 residues of the carboxyl terminus. Comparison of peptides with the inferred amino acid sequence of the E. coli K-12 enzyme has helped determine the sequence of the amino terminal 59 residues; only two differences between the sequences are noted in this region

Abnormal nuclear envelope in the cerebellar Purkinje cells and impaired motor learning in DYT11 myoclonus-dystonia mouse models.

Science.gov (United States)

Yokoi, Fumiaki; Dang, Mai T; Yang, Guang; Li, Jindong; Doroodchi, Atbin; Zhou, Tong; Li, Yuqing

2012-02-01

Myoclonus-dystonia (M-D) is a movement disorder characterized by myoclonic jerks with dystonia. DYT11 M-D is caused by mutations in SGCE which codes for ɛ-sarcoglycan. SGCE is maternally imprinted and paternally expressed. Abnormal nuclear envelope has been reported in mouse models of DYT1 generalized torsion dystonia. However, it is not known whether similar alterations occur in DYT11 M-D. We developed a mouse model of DYT11 M-D using paternally inherited Sgce heterozygous knockout (Sgce KO) mice and reported that they had myoclonus and motor coordination and learning deficits in the beam-walking test. However, the specific brain regions that contribute to these phenotypes have not been identified. Since ɛ-sarcoglycan is highly expressed in the cerebellar Purkinje cells, here we examined the nuclear envelope in these cells using a transmission electron microscope and found that they are abnormal in Sgce KO mice. Our results put DYT11 M-D in a growing family of nuclear envelopathies. To analyze the effect of loss of ɛ-sarcoglycan function in the cerebellar Purkinje cells, we produced paternally inherited cerebellar Purkinje cell-specific Sgce conditional knockout (Sgce pKO) mice. Sgce pKO mice showed motor learning deficits, while they did not show abnormal nuclear envelope in the cerebellar Purkinje cells, robust motor deficits, or myoclonus. The results suggest that ɛ-sarcoglycan in the cerebellar Purkinje cells contributes to the motor learning, while loss of ɛ-sarcoglycan in other brain regions may contribute to nuclear envelope abnormality, myoclonus and motor coordination deficits. Copyright © 2011 Elsevier B.V. All rights reserved.

Molecular and Phenotypic Characterization of Escherichia coli O26:H8 among Diarrheagenic E. coli O26 Strains Isolated in Brazil

Science.gov (United States)

Piazza, Roxane M. F.; Delannoy, Sabine; Fach, Patrick; Saridakis, Halha O.; Pedroso, Margareth Z.; Rocha, Letícia B.; Gomes, Tânia A. T.; Vieira, Mônica A. M.; Beutin, Lothar

2013-01-01

Escherichia coli strains of serogroup O26 comprise two distinct groups of pathogens, characterized as enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC). Among the several genes related to type III secretion system-secreted effector proteins, espK was found to be highly specific for EHEC O26:H11 and its stx-negative derivative strains isolated in European countries. E. coli O26 strains isolated in Brazil from infant diarrhea, foods, and the environment have consistently been shown to lack stx genes and are thus considered atypical EPEC. However, no further information related to their genetic background is known. Therefore, in this study, we aimed to discriminate and characterize these Brazilian O26 stx-negative strains by phenotypic, genetic, and biochemical approaches. Among 44 isolates confirmed to be O26 isolates, most displayed flagellar antigen H11 or H32. Out of the 13 nonmotile isolates, 2 tested positive for fliCH11, and 11 were fliCH8 positive. The identification of genetic markers showed that several O26:H11 and all O26:H8 strains tested positive for espK and could therefore be discriminated as EHEC derivatives. The presence of H8 among EHEC O26 and its stx-negative derivative isolates is described for the first time. The interaction of three isolates with polarized Caco-2 cells and with intestinal biopsy specimen fragments ex vivo confirmed the ability of the O26 strains analyzed to cause attaching-and-effacing (A/E) lesions. The O26:H32 strains, isolated mostly from meat, were considered nonvirulent. Knowledge of the virulence content of stx-negative O26 isolates within the same serotype helped to avoid misclassification of isolates, which certainly has important implications for public health surveillance. PMID:23974139

Genomic Comparison of Escherichia coli K1 Strains Isolated from the Cerebrospinal Fluid of Patients with Meningitis †

OpenAIRE

Yao, Yufeng; Xie, Yi; Kim, Kwang Sik

2006-01-01

Escherichia coli is a major cause of enteric/diarrheal diseases, urinary tract infections, and sepsis. E. coli K1 is the leading gram-negative organism causing neonatal meningitis, but the microbial basis of E. coli K1 meningitis is incompletely understood. Here we employed comparative genomic hybridization to investigate 11 strains of E. coli K1 isolated from the cerebrospinal fluid (CSF) of patients with meningitis. These 11 strains cover the majority of common O serotypes in E. coli K1 iso...

Staranje cevi uparjalnikov v Jedrski elektrarni Krško: Aging of tubes in the Krško nuclear power plant's steam generators:

OpenAIRE

Androjna, Ferdo; Cizelj, Leon

2000-01-01

The paper reviews the domestic efforts devoted to the safe and reliable operation of the Krško nuclear power plant (NPP) at full power, close to the design limit of the steam generators (18% of plugged tubes) for a full decade. This includes an overview of the recent status and history of the degradation processes, discussion of repair criteria, defining the acceptable size of defects and selected results from safety analyses supporting the operation of degraded steam generator (SG) tubes. It...

Saccharomyces genome database informs human biology

OpenAIRE

Skrzypek, Marek S; Nash, Robert S; Wong, Edith D; MacPherson, Kevin A; Hellerstedt, Sage T; Engel, Stacia R; Karra, Kalpana; Weng, Shuai; Sheppard, Travis K; Binkley, Gail; Simison, Matt; Miyasato, Stuart R; Cherry, J Michael

2017-01-01

Abstract The Saccharomyces Genome Database (SGD; http://www.yeastgenome.org) is an expertly curated database of literature-derived functional information for the model organism budding yeast, Saccharomyces cerevisiae. SGD constantly strives to synergize new types of experimental data and bioinformatics predictions with existing data, and to organize them into a comprehensive and up-to-date information resource. The primary mission of SGD is to facilitate research into the biology of yeast and...

Antimicrobial Resistant Pattern of Escherichia Coli Strains Isolated from Pediatric Patients in Jordan

Directory of Open Access Journals (Sweden)

Mohammad Alshara

2011-05-01

Full Text Available The present study was conducted to investigate antimicrobial resistant pattern of Escherichia coli (E. coli strains isolated from clinical specimens of Jordanian pediatric patients during the period from January to December 2008. A total of 444 E. coli strains were isolated from clinical specimens and tested for their susceptibility to different antimicrobial drugs. Overall, high resistance rate was observed for ampicillin (84%, followed by amoxicillin-clavulanic acid (74.3%, cotrimoxazole (71%, nalidixic acid (47.3%, cephalothin (41%. Lower resistance rates were observed for amikacin (0% followed by Cefotaxime (11%, Ceftriaxone (11.7%, ciprofloxacin (14.5%, Norfloxacin (16.5%, gentamicin (17.3% cephalexin (20.9%, Ceftazidime (22.5%, cefixime (29.6%, and cefaclor (32.8%. Ampicillin, amoxicillin-clavulanic acid and cotrimoxazole were found to be ineffective at in vitro inhibition of the E. coli of pediatric origin. Amikacin was highly effective for E. coli with susceptibility rate of 100%. The majority of E. coli strains were susceptible to third generation cephalosporins and fluoroquinolones.

Effects of Infrared Optical Trapping on Saccharomyces cerevisiae in a Microfluidic System

Czech Academy of Sciences Publication Activity Database

Pilát, Zdeněk; Jonáš, A.; Ježek, Jan; Zemánek, Pavel

2017-01-01

Roč. 17, NOV (2017), s. 1-12, č. článku 2640. ISSN 1424-8220 R&D Projects: GA MŠk(CZ) LO1212; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 Keywords : optical trapping * microfluidics * phototoxicity * laser * Saccharomyces cerevisiae Subject RIV: BH - Optics, Masers, Lasers OBOR OECD: Optics (including laser optics and quantum optics) Impact factor: 2.677, year: 2016 http://www.mdpi.com/1424-8220/17/11/2640

Biotechnology of non-Saccharomyces yeasts--the ascomycetes.

Science.gov (United States)

Johnson, Eric A

2013-01-01

Saccharomyces cerevisiae and several other yeast species are among the most important groups of biotechnological organisms. S. cerevisiae and closely related ascomycetous yeasts are the major producer of biotechnology products worldwide, exceeding other groups of industrial microorganisms in productivity and economic revenues. Traditional industrial attributes of the S. cerevisiae group include their primary roles in food fermentations such as beers, cider, wines, sake, distilled spirits, bakery products, cheese, sausages, and other fermented foods. Other long-standing industrial processes involving S. cerevisae yeasts are production of fuel ethanol, single-cell protein (SCP), feeds and fodder, industrial enzymes, and small molecular weight metabolites. More recently, non-Saccharomyces yeasts (non-conventional yeasts) have been utilized as industrial organisms for a variety of biotechnological roles. Non-Saccharomyces yeasts are increasingly being used as hosts for expression of proteins, biocatalysts and multi-enzyme pathways for the synthesis of fine chemicals and small molecular weight compounds of medicinal and nutritional importance. Non-Saccharomyces yeasts also have important roles in agriculture as agents of biocontrol, bioremediation, and as indicators of environmental quality. Several of these products and processes have reached commercial utility, while others are in advanced development. The objective of this mini-review is to describe processes currently used by industry and those in developmental stages and close to commercialization primarily from non-Saccharomyces yeasts with an emphasis on new opportunities. The utility of S. cerevisiae in heterologous production of selected products is also described.

Zdeněk Kožmín a strukturalismus

Czech Academy of Sciences Publication Activity Database

Sládek, Ondřej

2016-01-01

Roč. 19, č. 2 (2016), s. 32-48 ISSN 1213-2144 Institutional support: RVO:68378068 Keywords : Kožmín, Zdeněk * Mukařovský, Jan * Structural ism * Structural poetics and aesthetics * TheoryofInterpretation * Philosophy and Literature * Mukařovský, Jan * Structural ism * Structural poetics and aesthetics * Theory of Interpretation * Philosophy and Literature Subject RIV: AJ - Letters, Mass-media, Audiovision

The experiences to improve plant performance and reliability of Ko-Ri nuclear power plants

International Nuclear Information System (INIS)

Kang, Ho Weon

1998-01-01

This paper provides a discussion of the lessons learned from operational experience and the future plans to improve performance of the Ko-Ri plant. To operate nuclear power plants safely with good performance is the only way to mitigate the negative image of nuclear power generation to the public and to enhance the economical benefit compared to other electrical generation method. Therefore, in a continuous effort to overcome a negative challenge from outside, we have driven an aggressive 'OCTF' campaign as part of safety. As a result of our efforts, the following remarkable achievements have been accomplished. (1) 3 times of OCTF during recent three years (2) Selected twice as a top notch power plant on the list of NEI magazine in terms of plant capacity factor (3) No scram recorded in 1997 for all 4 units at Ko-Ri site. Ko-Ri is now undergoing the large scale plant betterment projects for retaking-off our operating performance to the level of new challenge target. Such improvement of critical components in the reactor coolant system and turbine system greatly contribute to increase the safety and reliability of the plant and to shortening of the planned outage period as well as to reduction of radiation exposure and radwaste. (Cho, G. S.). 5 tabs., 10 figs

Saccharomyces cerevisiae and non-Saccharomyces yeasts in grape varieties of the São Francisco Valley

Directory of Open Access Journals (Sweden)

Camila M.P.B.S. de Ponzzes-Gomes

2014-06-01

Full Text Available The aims of this work was to characterise indigenous Saccharomyces cerevisiae strains in the naturally fermented juice of grape varieties Cabernet Sauvignon, Grenache, Tempranillo, Sauvignon Blanc and Verdejo used in the São Francisco River Valley, northeastern Brazil. In this study, 155 S. cerevisiae and 60 non-Saccharomyces yeasts were isolated and identified using physiological tests and sequencing of the D1/D2 domains of the large subunit of the rRNA gene. Among the non-Saccharomyces species, Rhodotorula mucilaginosa was the most common species, followed by Pichia kudriavzevii, Candida parapsilosis, Meyerozyma guilliermondii, Wickerhamomyces anomalus, Kloeckera apis, P. manshurica, C. orthopsilosis and C. zemplinina. The population counts of these yeasts ranged among 1.0 to 19 x 10(5 cfu/mL. A total of 155 isolates of S. cerevisiae were compared by mitochondrial DNA restriction analysis, and five molecular mitochondrial DNA restriction profiles were detected. Indigenous strains of S. cerevisiae isolated from grapes of the São Francisco Valley can be further tested as potential starters for wine production.

Gains and Losses of Transcription Factor Binding Sites in Saccharomyces cerevisiae and Saccharomyces paradoxus

Science.gov (United States)

Schaefke, Bernhard; Wang, Tzi-Yuan; Wang, Chuen-Yi; Li, Wen-Hsiung

2015-01-01

Gene expression evolution occurs through changes in cis- or trans-regulatory elements or both. Interactions between transcription factors (TFs) and their binding sites (TFBSs) constitute one of the most important points where these two regulatory components intersect. In this study, we investigated the evolution of TFBSs in the promoter regions of different Saccharomyces strains and species. We divided the promoter of a gene into the proximal region and the distal region, which are defined, respectively, as the 200-bp region upstream of the transcription starting site and as the 200-bp region upstream of the proximal region. We found that the predicted TFBSs in the proximal promoter regions tend to be evolutionarily more conserved than those in the distal promoter regions. Additionally, Saccharomyces cerevisiae strains used in the fermentation of alcoholic drinks have experienced more TFBS losses than gains compared with strains from other environments (wild strains, laboratory strains, and clinical strains). We also showed that differences in TFBSs correlate with the cis component of gene expression evolution between species (comparing S. cerevisiae and its sister species Saccharomyces paradoxus) and within species (comparing two closely related S. cerevisiae strains). PMID:26220934

Metabolic Engineering of Probiotic Saccharomyces boulardii

OpenAIRE

Liu, Jing-Jing; Kong, In Iok; Zhang, Guo-Chang; Jayakody, Lahiru N.; Kim, Heejin; Xia, Peng-Fei; Kwak, Suryang; Sung, Bong Hyun; Sohn, Jung-Hoon; Walukiewicz, Hanna E.; Rao, Christopher V.; Jin, Yong-Su

2016-01-01

Saccharomyces boulardii is a probiotic yeast that has been used for promoting gut health as well as preventing diarrheal diseases. This yeast not only exhibits beneficial phenotypes for gut health but also can stay longer in the gut than Saccharomyces cerevisiae. Therefore, S. boulardii is an attractive host for metabolic engineering to produce biomolecules of interest in the gut. However, the lack of auxotrophic strains with defined genetic backgrounds has hampered the use of this strain for...

[Rehabilitation in the View of Small and Midsized Enterprises: Knowledge, Appreciation and Possibilities to Cooperate - Results of the KoRB-Project

DEFF Research Database (Denmark)

Hesse, B.; Heuer, J.; Gebauer, E.

2008-01-01

The experiences of small and midsized enterprises (SME) with medical rehabilitation were investigated in the KoRB-Study (KoRB= KOoperation Rehabilitation und Betrieb) for the first time. Opportunities for a targeted cooperation between the statutory pension insurance and these enterprises were...

Saccharomyces cerevisiae GTPase complex: Gtr1p-Gtr2p regulates cell-proliferation through Saccharomyces cerevisiae Ran-binding protein, Yrb2p

International Nuclear Information System (INIS)

Wang Yonggang; Nakashima, Nobutaka; Sekiguchi, Takeshi; Nishimoto, Takeharu

2005-01-01

A Gtr1p GTPase, the GDP mutant of which suppresses both temperature-sensitive mutants of Saccharomyces cerevisiae RanGEF/Prp20p and RanGAP/Rna1p, was presently found to interact with Yrb2p, the S. cerevisiae homologue of mammalian Ran-binding protein 3. Gtr1p bound the Ran-binding domain of Yrb2p. In contrast, Gtr2p, a partner of Gtr1p, did not bind Yrb2p, although it bound Gtr1p. A triple mutant: yrb2Δ gtr1Δ gtr2Δ was lethal, while a double mutant: gtr1Δ gtr2Δ survived well, indicating that Yrb2p protected cells from the killing effect of gtr1Δ gtr2Δ. Recombinant Gtr1p and Gtr2p were purified as a complex from Escherichia coli. The resulting Gtr1p-Gtr2p complex was comprised of an equal amount of Gtr1p and Gtr2p, which inhibited the Rna1p/Yrb2 dependent RanGAP activity. Thus, the Gtr1p-Gtr2p cycle was suggested to regulate the Ran cycle through Yrb2p

Inner boundary reform in Košice: The most fragmented city in Slovakia

Directory of Open Access Journals (Sweden)

Csachová Stela

2016-01-01

Full Text Available In the first part of the paper the aim is to provide the reader with a theoretical insight into models of government structure at the local level and provide evidence from European countries that underwent reforms during the second half of the 20th century. In the Slovak Republic (Slovakia the renewal of local self-government in 1990 brought changes to the administrative boundary map with more than 200 municipalities reestablished bringing the current number to 2,929 local administrative units. This has resulted in some discussion regarding a reduction in the number of local government units in this country. The second part of the paper is focused on Košice; the second largest city in Slovakia with 234,000 in­habitants. The city is organised at two administrative levels; one covering the whole city, the other covering 22 wards. When compared to cities of a similar size, Košice has been criticised for its high number of city wards and councillors, with the concomitant financial inefficiency it can bring. There has been an ongoing debate on reducing their number since 2010. The paper reports on the search for an admin­istrative structure that would best tailor to the citizens' needs and provide effective management in Košice. It uses traditional methods of qualitative research (historical method, content analysis and syn­thesis accompanied by statistical data organised in tables and figures.

[Andreeva, E. and Belobrovtseva, I. "Tol'ko Vy poimete sleduiushchii tekst..."] / Avril Pyman

Index Scriptorium Estoniae

Pyman, Avril

2014-01-01

Arvustus: Andreeva, E. and Belobrovtseva, I. "Tol'ko Vy poimete sleduiushchii tekst...". Perepiska N. E. Andreeva i L. F. Zurova. Baltiiskii arkhiv: Russkaia kult'tura v Pribaltike, 13. Tallinna Ülikooli Slaavi Keelte ja Kultuuride Instituut, Tallinn, 2013

Functional consequences of brain glycogen deficiency on the sleep-wake cycle regulation in PTG-KO mice

KAUST Repository

Burlet-Godinot, S.

2017-12-31

Introduction: In the CNS, glycogen is mainly localized in astrocytes where its levels are linked to neuronal activity. Astrocytic glycogen synthesis is regulated by glycogen synthase (GS) activity that is positively controlled by protein targeting to glycogen (PTG) expression levels. Although the role of glycogen in sleep/wake regulation is still poorly understood, we have previously demonstrated that, following a 6 hour gentle sleep deprivation (GSD), PTG mRNA expression and GS activity increased in the brain in mice while glycogen levels were paradoxically maintained and not affected. In order to gain further insight on the role of PTG in this process, we studied the sleep/wake cycle parameters in PTG knockout (PTG-KO) mice under baseline conditions and after a 6 hour GSD. Glycogen levels as well as mRNAs expression of genes related to energy metabolism were also determined in several brain areas. Materials and methods: Adult male C57BL/6J (WT) and PTG-KO mice were sleep-recorded under baseline conditions (24 h recordings, 12 h light/dark cycle) and following 6 hours GSD from ZT00 to ZT06. Vigilance states were visually scored (4 s temporal window). Spectral analysis of the EEG signal was performed using a discrete Fourier transformation. Glycogen measurements and gene expression analysis were assessed using a biochemical assay and quantitative RT-PCR respectively, on separate cohorts in WT vs PTG-KO mice at the end of the 6 hours GSD or in control animals (CTL) in different brain structures. Results: Quantitative analysis of the sleep/wake cycle under baseline conditions did not reveal major differences between the WT and the PTG-KO mice. However, during the dark period, the PTG-KO mice showed a significant increase in the number of wake and slow wave sleep episodes (respectively +26.5±8% and +26.1±8%; p< 0.05) together with a significant shortening in their duration (-21.6±7.2% and -14.3±2.8%; p< 0.01). No such quantitative changes were observed during

Infection with koala retrovirus subgroup B (KoRV-B), but not KoRV-A, is associated with chlamydial disease in free-ranging koalas (Phascolarctos cinereus)

OpenAIRE

Waugh, Courtney A.; Hanger, Jonathan; Loader, Joanne; King, Andrew; Hobbs, Matthew; Johnson, Rebecca; Timms, Peter

2017-01-01

The virulence of chlamydial infection in wild koalas is highly variable between individuals. Some koalas can be infected (PCR positive) with Chlamydia for long periods but remain asymptomatic, whereas others develop clinical disease. Chlamydia in the koala has traditionally been studied without regard to coinfection with other pathogens, although koalas are usually subject to infection with koala retrovirus (KoRV). Retroviruses can be immunosuppressive, and there is evidence of an immunosuppr...

Review of Saccharomyces boulardii as a treatment option in IBD

DEFF Research Database (Denmark)

Sivananthan, Kavitha; Petersen, Andreas Munk

2018-01-01

CONTEXT: Review of the yeast Saccharomyces boulardii as a treatment option for the inflammatory bowel diseases (IBD) ulcerative colitis and Crohn's disease. OBJECTIVE: IBD is caused by an inappropriate immune response to gut microbiota. Treatment options could therefore be prebiotics, probiotics......, antibiotics and/or fecal transplant. In this review, we have looked at the evidence for the yeast S. boulardii as a treatment option. MATERIAL AND METHODS: Searches in PubMed and the Cochrane Library with the MeSH words 'Saccharomyces boulardii AND IBD', 'Saccharomyces boulardii AND Inflammatory Bowel Disease....... Saccharomyces boulardii is, however, a plausible treatment option in the future, but more placebo-controlled clinical studies on both patients with ulcerative colitis and Crohn's disease are needed....

CULTIVO EXPERIMENTAL DEL CLADOCERO Moina sp ALIMENTADO CON Ankistrodesmus sp y Saccharomyces cereviseae

Directory of Open Access Journals (Sweden)

Martha Prieto

2006-05-01

Full Text Available Saccharomyces cereviseae. Materiales y métodos. Fueron realizados cultivos experimentales de la cepade cladóceros Moina sp en el laboratorio de Alimento Vivo de la Universidad de Córdoba, bajo condicionescontroladas de temperatura (22oC, pH (7.6, intensidad lumínica (2000 lux y aireación. Se emplearondos dietas de alimento como tratamiento (Ankistrodesmus y Ankistrodesmus mas Saccharomyces cereviseaeen concentración de 40 x 105 cel.ml-1 cada uno, para determinar su efecto sobre el desempeño de lapoblación. Diariamente se registró la densidad poblacional y celular. Resultados. Se obtuvieron diferenciassignificativas para el efecto de las dietas sobre el crecimiento poblacional de los cladóceros, los individuosalimentados con Ankistrodesmus sp + Saccharomyces cereviseae alcanzaron una densidad de 12.3a±0.30 org/ml-1 presentando mayor tasa instantánea de crecimiento (K 0.36 ± 0.002, el menor tiempode duplicación (td 1.94 ± 0.012 y el mayor rendimiento (r 1,1 ± 0.07 clad.ml-1.día-1. Conclusión. Loscladóceros por sus características de crecimiento en cultivo, presentan adaptación favorable a lascondiciones de manejo para la producción de biomasas potencialmente útiles como partícula nutritivacon fines acuícolas.

Gains and Losses of Transcription Factor Binding Sites in Saccharomyces cerevisiae and Saccharomyces paradoxus.

Science.gov (United States)

Schaefke, Bernhard; Wang, Tzi-Yuan; Wang, Chuen-Yi; Li, Wen-Hsiung

2015-07-27

Gene expression evolution occurs through changes in cis- or trans-regulatory elements or both. Interactions between transcription factors (TFs) and their binding sites (TFBSs) constitute one of the most important points where these two regulatory components intersect. In this study, we investigated the evolution of TFBSs in the promoter regions of different Saccharomyces strains and species. We divided the promoter of a gene into the proximal region and the distal region, which are defined, respectively, as the 200-bp region upstream of the transcription starting site and as the 200-bp region upstream of the proximal region. We found that the predicted TFBSs in the proximal promoter regions tend to be evolutionarily more conserved than those in the distal promoter regions. Additionally, Saccharomyces cerevisiae strains used in the fermentation of alcoholic drinks have experienced more TFBS losses than gains compared with strains from other environments (wild strains, laboratory strains, and clinical strains). We also showed that differences in TFBSs correlate with the cis component of gene expression evolution between species (comparing S. cerevisiae and its sister species Saccharomyces paradoxus) and within species (comparing two closely related S. cerevisiae strains). © The Author(s) 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

Tolerance to winemaking stress conditions of Patagonian strains of Saccharomyces eubayanus and Saccharomyces uvarum.

Science.gov (United States)

Origone, A C; Del Mónaco, S M; Ávila, J R; González Flores, M; Rodríguez, M E; Lopes, C A

2017-08-01

Evaluating the winemaking stress tolerance of a set of both Saccharomyces eubayanus and Saccharomyces uvarum strains from diverse Patagonian habitats. Yeast strains growth was analysed under increasing ethanol concentrations; all of them were able to grow until 8% v/v ethanol. The effect of different temperature and pH conditions as well as at SO 2 and hexose concentrations was evaluated by means of a central composite experimental design. Only two S. uvarum strains (NPCC 1289 and 1321) were able to grow in most stress conditions. Kinetic parameters analysed (μ max and λ) were statistically affected by temperature, pH and SO 2 , but not influenced by sugar concentration. The obtained growth model was used for predicting optimal growth conditions for both strains: 20°C, 0% w/v SO 2 and pH 4·5. Strains from human-associated environments (chichas) presented the highest diversity in the response to different stress factors. Two S. uvarum strains from chichas demonstrated to be the most tolerant to winemaking conditions. This work evidenced the potential use of two S. uvarum yeast strains as starter cultures in wines fermented at low temperatures. Saccharomyces eubayanus was significantly affected by winemaking stress conditions, limiting its use in this industry. © 2017 The Society for Applied Microbiology.

The experiences to improve plant performance and reliability of Ko-Ri nuclear power plants

Energy Technology Data Exchange (ETDEWEB)

Kang, Ho Weon [Korea Electric Power Corp. Ko-Ri nuclear power division, Ko-Ri (Korea, Republic of)

1998-07-01

This paper provides a discussion of the lessons learned from operational experience and the future plans to improve performance of the Ko-Ri plant. To operate nuclear power plants safely with good performance is the only way to mitigate the negative image of nuclear power generation to the public and to enhance the economical benefit compared to other electrical generation method. Therefore, in a continuous effort to overcome a negative challenge from outside, we have driven an aggressive 'OCTF' campaign as part of safety. As a result of our efforts, the following remarkable achievements have been accomplished. (1) 3 times of OCTF during recent three years (2) Selected twice as a top notch power plant on the list of NEI magazine in terms of plant capacity factor (3) No scram recorded in 1997 for all 4 units at Ko-Ri site. Ko-Ri is now undergoing the large scale plant betterment projects for retaking-off our operating performance to the level of new challenge target. Such improvement of critical components in the reactor coolant system and turbine system greatly contribute to increase the safety and reliability of the plant and to shortening of the planned outage period as well as to reduction of radiation exposure and radwaste. (Cho, G. S.). 5 tabs., 10 figs.

Improvement of ethanol-tolerance of haploid Saccharomyces diastaticus

International Nuclear Information System (INIS)

Song, S.H.; Kim, K.; Lee, M.W.

1994-01-01

Several mutation procedures have been compared to obtain an ethanol-tolerant Saccharomyces diastaticus strain secreting glucoamylase. These procedures include spontaneous mutation, EMS treatment, UV irradiation, and combination of EMS treatment and UV irradiation. All these methods were followed by adaptation of the yeast cells to gradually higher ethanol concentration. Among these procedures, the combined method of EMS treatment and UV irradiation gave the promising result, i.e. the ethanol tolerance of the yeast increased from 11.5%(v/v) to 14.0%(v/v). Respiratory deficient petite mutants of industrial and ethanol-tolerant yeast strains have been isolated and hybridized with haploid S. diastaticus strains. The resulting hybrids showed increased ethanol tolerance and starch-fermentability

Mechanisms of uv mutagenesis in yeast and E. coli

International Nuclear Information System (INIS)

Lawrence, C.; Christensen, R.; Christensen, J.R.; O'Brien, T.

1983-01-01

Experiments investigating ultraviolet light mutagenesis in either bakers' yeast, Saccharomyces cerevisiae, or E. coli have led to the following conclusions. First, cyclobutane pyrimidine dimers cause most mutations in both organisms; pyrimidine adducts, such as PyC, can account at best for only a small proportion. 86 percent of forward mutations induced at the E. coli lacI locus can be abolished by photoreactivation under conditions which do not alter the level of recA induction. About 75 percent of the forward mutations induced at the CAN1 locus of yeast could be removed by photoreactivation, a value that lies within the range observed previously for the reversion of CYC1 alleles (60 percent - 97 percent). Second, about 10 percent of the lacI forward mutations are untargeted, a smaller fraction than found previously for cycl-91 reversion in yeast. It is not yet clear whether the two species are really different in this respect, of whether the cycl-91 reversion site is a typical of the yeast genome at large. Third, analysis of reversion frequencies of 20 mutant alleles suggests that about 10 to 25 percent of all replication errors produced by mutagenic mechanisms in uv-irradiated yeast involve additions or deletions of base-pairs, indicating that error-prone repair does not just produce substitutions. Last, the REV1 locus in yeast is concerned with the induction of frameshift mutations at some, but not all, genetic sites, just as found previously for substitution mutations. The function of the REV3 gene is more widely, though not universally, required while the function of the RAD6 gene, like that of the recA locus in E. coli, appears to be necessary for all kinds of uv mutagenesis. E coli genes comparable to REV1 and REV3 have not yet been described; conversely, there does not yet appear to be a yeast equivalent of umuC

Mechanisms of uv mutagenesis in yeast and E. coli

International Nuclear Information System (INIS)

Lawrence, C.; Christensen, R.; Christensen, J.R.; O'Brien, T.

1983-01-01

Experiments investigating ultraviolet light mutagenesis in either bakers' yeast, Saccharomyces cerevisiae, or E. coli have led to the following conclusions. First, cyclobutane pyrimidine dimers cause most mutations in both organisms; pyrimidine adducts, such as PyC, can account at best for only a small proportion. Eighty-six percent of forward mutations induced at the E. coli lacI locus can be abolished by photoreactivation under conditions which do not alter the level of recA induction. About 75 percent of the forward mutations induced at the CAN1 locus of yeast could be removed by photoreactivation, a value that lies within the range observed previously for the reversion of CYC1 alleles (60 percent - 97 percent). Second, about 10 percent of the lacI forward mutations are untargeted, a smaller fraction than found previously for cycl1-91 reversion in yeast. It is not yet clear whether the two species are really different in this respect, or whether the cyc1-91 reversion site is atypical of the yeast genome at large. Third, analysis of reversion frequencies of 20 mutant alleles suggests that about 10 - 25 percent of all replication errors produced by mutagenic mechanisms in UV-irradiated yeast involve additions or deletions of base-pairs, indicating that error-prone repair does not just produce substitutions. Last, the REV1 locus in yeast is concerned with the induction of frameshift mutations at some, but not all, genetic sites, just as found previously for substitution mutations. The function of the REV3 gene is more widely, though not universally, required while the function of the RAD6 gene, like that of the recA locus in E. coli, appears to be necessary for all kinds of UV mutagenesis. E. coli genes comparable to REV1 and REV3 have not yet been described, conversely, there does not yet appear to be a yeast equivalent of umuC. 13 references, 4 tables

Maria Avdjuško väntab Priimäest filmi / Verni Leivak

Index Scriptorium Estoniae

Leivak, Verni, 1966-

2009-01-01

Näitlejanna debüteerib režissöörina, vändates lühimängufilmi, kus Linnar Priimägi peategelasena kehastab iseennast. Operaatoriks on Istvan Borbas Rootsist, näitlejateks veel M. Avdjuško ja Taavi Eelmaa

Professor Krystyna Kotełko and her contribution to the study of Proteus endotoxin.

Science.gov (United States)

Różalski, Antoni W

2018-04-01

Professor Krystyna Kotełko was working as a microbiologist at the University of Łódź (Poland). Her main object of study was the LPS (endotoxin) of opportunistic urinary pathogens from the genus Proteus. She demonstrated, for the first time, the presence of uronic acids and amino acids, as well as two heptoses (L- glycero-D- manno-heptose and D- glycero-D- manno-heptose) and hexosamines in Proteus LPS, and developed a classification scheme of the Proteus LPS into chemotypes. Prof Kotełko also initiated studies on the chemical structure of Proteus O-specific polysaccharide and investigations on the serological specificity of this part of LPS, as well its core region. She also analysed the virulence factors of these bacteria, such as haemolysin and invasiveness.

Zymogram profiling of superoxide dismutase and catalase activities allows Saccharomyces and non-Saccharomyces species differentiation and correlates to their fermentation performance.

Science.gov (United States)

Gamero-Sandemetrio, Esther; Gómez-Pastor, Rocío; Matallana, Emilia

2013-05-01

Aerobic organisms have devised several enzymatic and non-enzymatic antioxidant defenses to deal with reactive oxygen species (ROS) produced by cellular metabolism. To combat such stress, cells induce ROS scavenging enzymes such as catalase, peroxidase, superoxide dismutase (SOD) and glutathione reductase. In the present research, we have used a double staining technique of SOD and catalase enzymes in the same polyacrylamide gel to analyze the different antioxidant enzymatic activities and protein isoforms present in Saccharomyces and non-Saccharomyces yeast species. Moreover, we used a technique to differentially detect Sod1p and Sod2p on gel by immersion in NaCN, which specifically inhibits the Sod1p isoform. We observed unique SOD and catalase zymogram profiles for all the analyzed yeasts and we propose this technique as a new approach for Saccharomyces and non-Saccharomyces yeast strains differentiation. In addition, we observed functional correlations between SOD and catalase enzyme activities, accumulation of essential metabolites, such as glutathione and trehalose, and the fermentative performance of different yeasts strains with industrial relevance.

The essential oil of Amomum tsao-ko Crevost et Lemarie from Vietnam

NARCIS (Netherlands)

Dung, N.X.; Bien, Le Kiem; Leclercq, P.A.

1992-01-01

The essential oil of the seeds of A. tsao-ko of Vietnamese origin was analyzed by a combination of gas chromatog. and gas chromatog./mass spectrometry. Twenty-one components were identified, of which the major ones were 1,8-cineole (30.6%), 2-decenal (17.3%), geranial (10.6%), and neural (7.0%)

MicroRNA and Transcriptomic Profiling Showed miRNA-Dependent Impairment of Systemic Regulation and Synthesis of Biomolecules in Rag2 KO Mice.

Science.gov (United States)

Reza, Abu Musa Md Talimur; Choi, Yun-Jung; Kim, Jin-Hoi

2018-02-27

The Rag2 knockout (KO) mouse is a well-established immune-compromised animal model for biomedical research. A comparative study identified the deregulated expression of microRNAs (miRNAs) and messenger RNAs (mRNAs) in Rag2 KO mice. However, the interaction between deregulated genes and miRNAs in the alteration of systemic (cardiac, renal, hepatic, nervous, and hematopoietic) regulations and the synthesis of biomolecules (such as l-tryptophan, serotonin, melatonin, dopamine, alcohol, noradrenaline, putrescine, and acetate) are unclear. In this study, we analyzed both miRNA and mRNA expression microarray data from Rag2 KO and wild type mice to investigate the possible role of miRNAs in systemic regulation and biomolecule synthesis. A notable finding obtained from this analysis is that the upregulation of several genes which are target molecules of the downregulated miRNAs in Rag2 KO mice, can potentially trigger the degradation of l-tryptophan, thereby leading to the systemic impairment and alteration of biomolecules synthesis as well as changes in behavioral patterns (such as stress and fear responses, and social recognition memory) in Rag2 gene-depleted mice. These findings were either not observed or not explicitly described in other published Rag2 KO transcriptome analyses. In conclusion, we have provided an indication of miRNA-dependent regulations of clinical and pathological conditions in cardiac, renal, hepatic, nervous, and hematopoietic systems in Rag2 KO mice. These results may significantly contribute to the prediction of clinical disease caused by Rag2 deficiency.

MicroRNA and Transcriptomic Profiling Showed miRNA-Dependent Impairment of Systemic Regulation and Synthesis of Biomolecules in Rag2 KO Mice

Directory of Open Access Journals (Sweden)

Abu Musa Md Talimur Reza

2018-02-01

Full Text Available The Rag2 knockout (KO mouse is a well-established immune-compromised animal model for biomedical research. A comparative study identified the deregulated expression of microRNAs (miRNAs and messenger RNAs (mRNAs in Rag2 KO mice. However, the interaction between deregulated genes and miRNAs in the alteration of systemic (cardiac, renal, hepatic, nervous, and hematopoietic regulations and the synthesis of biomolecules (such as l-tryptophan, serotonin, melatonin, dopamine, alcohol, noradrenaline, putrescine, and acetate are unclear. In this study, we analyzed both miRNA and mRNA expression microarray data from Rag2 KO and wild type mice to investigate the possible role of miRNAs in systemic regulation and biomolecule synthesis. A notable finding obtained from this analysis is that the upregulation of several genes which are target molecules of the downregulated miRNAs in Rag2 KO mice, can potentially trigger the degradation of l-tryptophan, thereby leading to the systemic impairment and alteration of biomolecules synthesis as well as changes in behavioral patterns (such as stress and fear responses, and social recognition memory in Rag2 gene-depleted mice. These findings were either not observed or not explicitly described in other published Rag2 KO transcriptome analyses. In conclusion, we have provided an indication of miRNA-dependent regulations of clinical and pathological conditions in cardiac, renal, hepatic, nervous, and hematopoietic systems in Rag2 KO mice. These results may significantly contribute to the prediction of clinical disease caused by Rag2 deficiency.

Effect of Saccharomyces cerevisiae var. Boulardii and β-galactomannan oligosaccharide on porcine intestinal epithelial and dendritic cells challenged in vitro with Escherichia coli F4 (K88

Directory of Open Access Journals (Sweden)

Badia Roger

2012-01-01

Full Text Available Abstract Probiotic and prebiotics, often called "immune-enhancing" feed additives, are believed to deal with pathogens, preventing the need of an immune response and reducing tissue damage. In this study, we investigated if a recently developed β-galactomannan (βGM had a similar protective role compared to Saccharomyces cerevisiae var. Boulardii (Scb, a proven probiotic, in the context of enterotoxigenic Escherichia coli (ETEC infection. ETEC causes inflammation, diarrhea and intestinal damage in piglets, resulting in large economic loses worldwide. We observed that Scb and βGM products inhibited in vitro adhesion of ETEC on cell surface of porcine intestinal IPI-2I cells. Our data showed that Scb and βGM decreased the mRNA ETEC-induced gene expression of pro-inflammatory cytokines TNF-α, IL-6, GM-CSF and chemokines CCL2, CCL20 and CXCL8 on intestinal IPI-2I. Furthermore, we investigated the putative immunomodulatory role of Scb and βGM on porcine monocyte-derived dendritic cells (DCs per se and under infection conditions. We observed a slight up-regulation of mRNA for TNF-α and CCR7 receptor after co-incubation of DC with Scb and βGM. However, no differences were found in DC activation upon ETEC infection and Scb or βGM co-culture. Therefore, our results indicate that, similar to probiotic Scb, prebiotic βGM may protect intestinal epithelial cells against intestinal pathogens. Finally, although these products may modulate DC activation, their effect under ETEC challenge conditions remains to be elucidated.

Molecular characterization of Escherichia coli recovered from traditional milk products in Kashan, Iran

Directory of Open Access Journals (Sweden)

Farhad Sharafati Chaleshtori

2017-10-01

Full Text Available Aim: Shiga toxigenic Escherichia coli (STEC strains as emerging groups of foodborne pathogens are responsible for most foodborne illnesses. The aim of this study was to determine the antibiotic resistance pattern in STEC isolated from traditional milk products and their molecular characterization. Materials and Methods: A total of 116 samples were randomly purchased from local markets in Kashan, Iran, and evaluated for the occurrence of STEC by culturing and molecular methods. The antibiotic resistance of obtained isolates was determined by Kirby Bauer method. Furthermore, isolates were assayed for the presence of Shiga toxins (stx1 and stx2 and intimin gene (eae. Results: The incidence of E. coli in 60 ice cream, 30 yoghurt, and 26 cheese samples was 8.33%, 10%, and 11.54%, respectively. The findings showed that 11 out of 11 (100% E. coli had both stx1 and stx2 while eae gene was not found in E. coli isolated of traditional milk products. For E. coli strains carrying stx1 and stx2, highest antibiotic sensitive levels were related to trimethoprim/sulfamethoxazole, norfloxacin, chloramphenicol, and ciprofloxacin, respectively. Conclusion: The results showed relationship between the presence of virulence factors and antimicrobial resistance. These results can be used for further studies on STEC as an emerging foodborne pathogen.

TopBP1/Dpb11 binds DNA anaphase bridges to prevent genome instability

DEFF Research Database (Denmark)

Germann, Susanne Manuela; Schramke, Vera; Pedersen, Rune Troelsgaard

2014-01-01

yeast Saccharomyces cerevisiae and the avian DT40 cell line as model systems for studying DNA anaphase bridges and show that TopBP1/Dpb11 plays an evolutionarily conserved role in their metabolism. Together with the single-stranded DNA binding protein RPA, TopBP1/Dpb11 binds to UFBs, and depletion...

Probiotic Activity of Saccharomyces cerevisiae var. boulardii Against Human Pathogens

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Katarzyna Rajkowska

2012-01-01

Full Text Available Infectious diarrhoea is associated with a modification of the intestinal microflora and colonization of pathogenic bacteria. Tests were performed for seven probiotic yeast strains of Saccharomyces cerevisiae var. boulardii, designated for the prevention and treatment of diarrhoea. To check their possible effectiveness against diarrhoea of different etiologies, the activity against a variety of human pathogenic or opportunistic bacteria was investigated in vitro. In mixed cultures with S. cerevisiae var. boulardii, a statistically significant reduction was observed in the number of cells of Listeria monocytogenes, Pseudomonas aeruginosa and Staphylococcus aureus, by even 55.9 % in the case of L. monocytogenes compared with bacterial monocultures. The influence of yeasts was mostly associated with the shortening of the bacterial lag phase duration, more rapid achievement of the maximum growth rates, and a decrease by 4.4–57.1 % (L. monocytogenes, P. aeruginosa, or an increase by 1.4–70.6 % (Escherichia coli, Enterococcus faecalis, Salmonella Typhimurium in the exponential growth rates. Another issue included in the research was the ability of S. cerevisiae var. boulardii to bind pathogenic bacteria to its cell surface. Yeasts have shown binding capacity of E. coli, S. Typhimurium and additionally of S. aureus, Campylobacter jejuni and E. faecalis. However, no adhesion of L. monocytogenes and P. aeruginosa to the yeast cell wall was noted. The probiotic activity of S. cerevisiae var. boulardii against human pathogens is related to a decrease in the number of viable and active cells of bacteria and the binding capacity of yeasts. These processes may limit bacterial invasiveness and prevent bacterial adherence and translocation in the human intestines.

Kampen om den røde ko - dominerende temaer i dansk kvægavl i 1800-tallets slutning

DEFF Research Database (Denmark)

Gjerløff, Anne Katrine

2009-01-01

En analyse af kvægavlslitteratur fra 1800-tallets anden halvdel viser spændingerne mellem avlernes ønske om profit og hensynene til racerenhed og national og regional identitet. Ideen om den perfekte ko var omdiskuteret og modsætningerne mellem koens udseende og dens ydelse er tydelige i periodens...... bedømmelser af "den gode ko". Artiklen komkluserer at et fokus på opfattelser og brug af dyr kan give nye vinkler på traditionelle historiske emnefelter. Udgivelsesdato: april...

The Eschericia coli Growth Inhibition Activity of Some Fermented Medicinal Plant Leaf Extract from the Karo Highland, North Sumatra

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NOVIK NURHIDAYAT

2009-10-01

Full Text Available A lot of traditional medicinal plant has antibacterial acitivities. Most of these plants are freshly chewed or grounded and used directly to treat infectious bacterial deseases. However, some practices employ a traditionally spontaneous fermentation on boiled extracted leaf, root or other parts of the plant. This work reports a laboratory stimulated spontaneous fermentation of leaf extracts from selected medicinal plants collected from the Karo Higland. The spontaenous fermentation was stimulated to be carried out by the Acetobacter xylinum and Saccharomyces cerevisiae. The anti-infectious agent activity was assayed on the Eschericia coli growth inhibition. A complementary non fermented leaf extract was also made and assayed as a comparative measure. Indeed, the fermented leaf extract of bitter bush (Eupatorium pallescens, cacao (Theobroma cacao, avocado (Persia gratissima, passion fruit (Passiflora edulis, cassava (Cassava utillissima, diamond flower (Hedyotis corymbosa, periwinkle (Catharanthus roseus, and gandarusa (Justicia gendarussa have relatively higher anti-E.coli acitivity than those of non fermented ones. However, there were no anti-E.coli activity was detected in both fermented and non fermented leaf extract of the guava (Psidium guajava and common betel (Piper nigrum.

Transport systems of Ventricaria ventricosa: I/V analysis of both membranes in series as a function of [K(+)](o).

Science.gov (United States)

Beilby, M J; Bisson, M A

1999-09-01

The current-voltage (I/V) profiles of Ventricaria (formerly Valonia) membranes were measured at a range of external potassium concentrations, [K(+)](o), from 0.1 to 100 mm. The conductance-voltage (G/V) characteristics were computed to facilitate better resolution of the profile change with time after exposure to different [K(+)](o). The resistance-voltage (R/V) characteristics were computed to attempt resolution of plasmalemma and tonoplast. Four basic electrophysiological stages emerged: (1) Uniform low resistance between -60 and +60 mV after the cell impalement. (2) High resistance between +50 and +150 for [K(+)](o) from 0.1 to 1.0 mm and hypotonic media. (3) High resistance between -150 and -20 mV for [K(+)](o) of 10 mm (close to natural seawater) and hypertonic media. (4) High resistance between -150 and +170 mV at [K(+)](o) of 100 mm. The changes between these states were slow, requiring minutes to hours and sometimes exhibiting spontaneous oscillations of the membrane p.d. (potential difference). Our analysis of the I/V data supports a previous hypothesis, that Ventricaria tonoplast is the more resistive membrane containing a pump, which transports K(+) into the vacuole to regulate turgor. We associate state (1) with the plasmalemma conductance being dominant and the K(+) pump at the tonoplast short-circuited probably by a K(+) channel, state (2) with the K(+) pump "off" or short-circuited at p.d.s more negative than +50 mV, state (3) with the K(+) pump "on, " and state (4) with the pump dominant, but affected by high K(+). A model for the Ventricaria membrane system is proposed.

Genetic characterization of Shiga toxin-producing Escherichia coli O26:H11 strains isolated from animal, food, and clinical samples

Science.gov (United States)

Krüger, Alejandra; Lucchesi, Paula M. A.; Sanso, A. Mariel; Etcheverría, Analía I.; Bustamante, Ana V.; Burgán, Julia; Fernández, Luciana; Fernández, Daniel; Leotta, Gerardo; Friedrich, Alexander W.; Padola, Nora L.; Rossen, John W. A.

2015-01-01

The Shiga-toxin producing Escherichia coli (STEC) may cause serious illness in human. Here we analyze O26:H11 strains known to be among the most reported STEC strains causing human infections. Genetic characterization of strains isolated from animal, food, and clinical specimens in Argentina showed that most carried either stx1a or stx2a subtypes. Interestingly, stx2a-positive O26:H11 rarely isolated from cattle in other countries showed to be an important proportion of O26:H11 strains circulating in cattle and food in our region. Seventeen percent of the isolates harbored more than one gene associated with antimicrobial resistance. In addition to stx, all strains contained the virulence genes eae-β, tir, efa, iha, espB, cif, espA, espF, espJ, nleA, nleB, nleC, and iss; and all except one contained ehxA, espP, and cba genes. On the other hand, toxB and espI genes were exclusively observed in stx2-positive isolates, whereas katP was only found in stx1a-positive isolates. Our results show that O26:H11 STEC strains circulating in Argentina, including those isolated from humans, cattle, and meat products, present a high pathogenic potential, and evidence that cattle can be a reservoir of O26:H11 strains harboring stx2a. PMID:26539413

Citation analysis of The Korean Journal of Internal Medicine from KoMCI, Web of Science, and Scopus.

Science.gov (United States)

Huh, Sun

2011-03-01

The Korean Journal of Internal Medicine (KJIM) is the international journal published in English by the Korean Association of Internal Medicine. To understand the position of the journal in three different databases, the citation indicators were elucidated. From databases such as Korean Medical Citation Index (KoMCI), Web of Science, and Scopus, citation indicators such as the impact factor, SCImago journal rank (SJR), or Hirsch Index were calculated according to the year and the results were drawn. The KJIM 2010 impact factor increased to 0.623 in Web of Science. That of year 2009 in KoMCI was a 0.149. The 2009 SJR in Scopus was 0.073, with a ranking of 27/72 (37.5%) in the category of internal medicine and 414/1,618 (25.6%) in the category of medicine, miscellaneous. The Hirsch Index from KoMCI, Web of Science and Scopus were 5, 14, and 16, respectively. The KJIM is now cited more by international researchers than Korean researchers, indicating that the content of the journal is now valued at the international level.

ANALISIS CEMARAN BAKTERI Escherichia coli ANALISIS CEMARAN BAKTERI Escherichia coli ANALISIS CEMARAN BAKTERI Escherichia coli

OpenAIRE

ANGGREINI, RAHAYU

2015-01-01

2015 RAHAYU ANGGREINI coli Penelitian ini bertujuan untuk melakukan identifikasi cemaran bakteri E. coli O157:H7 pada daging sapi di kota Makassar. Sampel pada penelitian ini sebanyak 72 sampel Kata Kunci : Daging sapi, pasar tradisional, E. coli, E. coli O157:H7, kontaminasi bakteri, identifikasi E. coli O157:H7.

PROTEKSI VAKSIN MONOVALEN DAN KOKTAIL SEL UTUH TERHADAP KO-INFEKSI Mycobacterium fortuitum DAN Aeromonas hydrophila PADA IKAN GURAME, Osphronemus gouramy Uni

Directory of Open Access Journals (Sweden)

Uni Purwaningsih

2014-08-01

serangan agen patogen. Pemberian vaksin diharapkan dapat merangsang respons imun spesifik dan non spesifik pada ikan. Keberhasilan vaksinasi dipengaruhi oleh konsentrasi antigen, reaksi silang dan kompetisi di antara antigen yang berbeda. Penelitian ini bertujuan untuk menganalisis level proteksi vaksin monovalen dan koktail sel utuh terhadap ko-infeksi M. fortuitum dan A. hydrophila pada ikan gurame. Respons imun perlakuan vaksin monovalen maupun koktail pada gurame pasca ko-infeksi menunjukkan nilai parameter-parameter yaitu: hematokrit, hemoglobin, indeks fagositik, persentase fagositik, titer antibodi, uji respiratory burst (NBT, aktivitas komplemen, dan diferensial leukosit yang berbeda nyata (P<0,05 dibanding kontrol. Vaksin monovalen A. hydrophila menunjukkan nilai RPS sebesar 92,3% dan monovalen M. fortuitum sebesar 78,6% setelah ditantang dengan infeksi tunggal bakteri homolog namun menunjukkan proteksi yang rendah terhadap ko-infeksi. Vaksin koktail 50Mf :50Ah memberikan proteksi yang lebih baik dari vaksin monovalen pasca uji tantang dengan ko-infeksi bakteri M. fortuitum dan A. hydrophila.

Investigation of autonomous cell cycle oscillation in Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Hansen, Morten Skov

2007-01-01

Autonome Oscillationer i kontinuert kultivering af Saccharomyces cerevisiae Udgangspunktet for dette Ph.d. projekt var at søge at forstå, hvad der gør det muligt at opnå multiple statiske tilstande ved kontinuert kultivering af Saccharomyces cerevisiae med glukose som begrænsende substrat...

The CRO-1 gene of Saccharomyces cerevisiae controls mitotic crossing over, chromosomal stability and sporulation

International Nuclear Information System (INIS)

Esposito, M.S.; Maleas, D.T.; Bjornstad, K.A.; Holbrook, L.L.

1987-01-01

The properties of a novel temperature-sensitive recombination-defective mutant of Saccharomyces cerevisiae, cro1-1 is described. The cro1-1 mutant is the first instance of a rec mutation that reduces drastically the rates of spontaneous mitotic crossing-over events but not those of gene conversional events. The cro1-1 mutation thus provides evidence that mitotic crossing-over is dependent upon gene products that are not essential for gene conversional events. The cro1-1 mutation also results in enhanced mitotic-chromosomal instability and MATa/MATα cro1-1/cro1-1 mutants are sporulation deficient. These phenotypes indicate that the CRO1 gene modulates mitotic chromosomal integrity and is essential for normal meiosis. The cro1-1 mutant possesses Holliday junction resolvase activity, hence its recombinational defect does not involve failure to execute this putative final recombinational step. 7 refs., 1 fig., 5 tabs

Saccharomyces boulardii CNCM I-745 in different clinical conditions.

Science.gov (United States)

Dinleyici, Ener Cagri; Kara, Ates; Ozen, Metehan; Vandenplas, Yvan

2014-11-01

Saccharomyces boulardii is a well-known probiotic worldwide, and there are numerous studies including experimental and clinical trials in children and adults by the use of S. boulardii. The objective of the present report is to provide an update on the evidence for the efficacy of S. boulardii CNCM I-745 in different clinical conditions. Saccharomyces boulardii is one of the best-studied probiotics in acute gastroenteritis (AGE) and is shown to be safe and to reduce the duration of diarrhea and hospitalization by about 1 day. Saccharomyces boulardii is one of the recommended probiotics for AGE in children by European Society of Paediatric Infectious Diseases and European Society for Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN). Saccharomyces boulardii is also a recommended probiotic for the prevention of antibiotic-associated diarrhea (AAD), and a recent study showed promising results for the treatment of AAD in children. There is insufficient evidence to recommend the long-term use of S. boulardii in patients with irritable bowel syndrome. Although some clinical studies showed positive effects of S. boulardii on inflammation, there is no clinical evidence that S. boulardii is useful in inflammatory bowel disease. Saccharomyces boulardii could be used in patients needing Helicobacter pylori eradication because the S. boulardii improves compliance, decreases the side effects and moderately increases the eradication rate. There are new promising results (improving feeding tolerance, shorten the course of hyperbilirubinemia), but we do still not recommend the routine use of S. boulardii in newborns. Saccharomyces boulardii CNCM I-745 is a good example for the statement that each probiotic needs to be taxonomically characterized and its efficacy and safety should be documented individually in different clinical settings.

[A prospective control study of Saccharomyces boulardii in prevention of antibiotic-associated diarrhea in the older inpatients].

Science.gov (United States)

Zhang, D M; Xu, B B; Yu, L; Zheng, L F; Chen, L P; Wang, W

2017-06-01

Objective: To study the value of Saccharomyces boulardii for the prevention of antibiotic-associated diarrhea in older inpatients. Methods: A total of 163 older patients who were treated with wide-spectrum antibiotics at least three days during January 2014 to December 2015 were randomly divided into control and study group. In study group, 81 patients were administrated with oral Saccharomyces boulardii 500 mg twice a day for 21 days. The control group was of no intervention. Morbidity rate of antibiotic-associated diarrhea and Clostridium difficile -associated diarrhea, frequency and duration of diarrhea were recorded. Results: The incidence of antibiotic-associated diarrhea in study group was significantly lower than that in control group [14.8%(12/81) vs 28.0%(23/82), P 0.05] in two groups. The frequency and duration of diarrhea in the study group were significantly lower and shorter than those in control group[(4.3±1.7) times/day vs (6.9±2.0) times/day; (3.0±1.1) days vs (5.7±1.8) days, both P Saccharomyces boulardii may reduce the incidence of antibiotic-associated diarrhea therefore improving the symptom of diarrhea in older inpatients.

Inheritance and organisation of the mitochondrial genome differ between two Saccharomyces yeasts

DEFF Research Database (Denmark)

Petersen, Randi Føns; Langkjær, Rikke Breinhold; Hvidtfeldt, J.

2002-01-01

Petite-positive Saccharomyces yeasts can be roughly divided into the sensu stricto, including Saccharomyces cerevisiae, and sensu lato group, including Saccharomyces castellii; the latter was recently studied for transmission and the organisation of its mitochondrial genome. S. castellii mitochon......Petite-positive Saccharomyces yeasts can be roughly divided into the sensu stricto, including Saccharomyces cerevisiae, and sensu lato group, including Saccharomyces castellii; the latter was recently studied for transmission and the organisation of its mitochondrial genome. S. castellii...... mitochondrial molecules (mtDNA) carrying point mutations, which confer antibiotic resistance, behaved in genetic crosses as the corresponding point mutants of S. cerevisiae. While S. castellii generated spontaneous petite mutants in a similar way as S. cerevisiae, the petites exhibited a different inheritance...... pattern. In crosses with the wild type strains a majority of S. castellii petites was neutral, and the suppressivity in suppressive petites was never over 50%. The two yeasts also differ in organisation of their mtDNA molecules. The 25,753 bp sequence of S. castellii mtDNA was determined and the coding...

Saccharomyces cerevisiae var. boulardii fungemia following probiotic treatment

OpenAIRE

Appel-da-Silva, Marcelo C.; Narvaez, Gabriel A.; Perez, Leandro R.R.; Drehmer, Laura; Lewgoy, Jairo

2017-01-01

Probiotics are commonly prescribed as an adjuvant in the treatment of antibiotic-associated diarrhea caused by Clostridium difficile. We report the case of an immunocompromised 73-year-old patient on chemotherapy who developed Saccharomyces cerevisiae var. boulardii fungemia in a central venous catheter during treatment of antibiotic-associated pseudomembranous colitis with the probiotic Saccharomyces cerevisiae var. boulardii. Fungemia was resolved after interruption of probiotic administrat...

An apoptotic cell cycle mutant in Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Villadsen, Ingrid

1996-01-01

The simple eukaryote Saccharomyces cerevisiae has proved to be a useful organism for elucidating the mechanisms that govern cell cycle progression in eukaryotic cells. The excellent in vivo system permits a cell cycle study using temperature sensitive mutants. In addition, it is possible to study...... many genes and gene products from higher eukaryotes in Saccharomyces cerevisiae because many genes and biological processes are homologous or similar in lower and in higher eukaryotes. The highly developed methods of genetics and molecular biology greatly facilitates studies of higher eukaryotic...... processes.Programmmed cell death with apoptosis plays a major role in development and homeostatis in most, if not all, animal cells. Apoptosis is a morphologically distinct form of death, that requires the activation of a highly regulated suicide program. Saccharomyces cerevisiae provides a new system...

[Effects of non-saccharomyces albicans metabolic products on the proliferation of human umbilical vein endothelial cell ECV304].

Science.gov (United States)

Chen, Bin; Che, Tuanjie; Bai, Decheng; He, Xiangyi

2013-04-01

To evaluate the effects of non-Saccharomyces albicans metabolic products on the cell cycle distribution and proliferation of human umbilical vein endothelial cell ECV304 cells in vitro. The parallel dilution supernatant of Saccharomyces tropicalis, Saccharomyces krusei and Saccharomyces glabrata were prepared, and 1, 4, 16-fold(s) diluted concentration and control group were set up. The line of human umbilical vein endothelial cell ECV304 was cultured in vitro and treated by non-Saccharomyces albicans supernatant. The proliferous effect of ECV304 induced by non-Saccharomyces albicans supernatant after 24, 48, 72 h was detected by the methods of MTT, and the changes of cell density and cycle after 48 h were investigated by inverted microscope and flow cytometry. At the 24th hour, all of the higher concentration (1-fold) of non-Saccharomyces albicans supernatant and the 4-folds diluted Saccharomyces krusei could promote ECV304 proliferation(P Saccharomyces albicans supernatant at 48h and 72th hour, Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant significantly increased proliferation rate of ECV304, while Saccharomyces tropicalis supernatant group showed no significant change no matter which concentration was tested. At 48th hour after adding the non-Saccharomyces albicans supernatant, the ECV304 cells density treated by Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant were significantly higher under the inverted microscope. The G0/G1 population of ECV304 cells decreased while cell proliferation index (PI) increased after incubated with Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant for 48 hours (P Saccharomyces tropicalis group showed no significant change (P > 0.05). The metabolic products of Sacharoymces krusei and Saccharomyces glabrata could induce proliferation of ECV304 cell, which suggests non-Saccharomyces albicans should be undergone more attention clinically in detection and treatment.

Non-Saccharomyces in Wine: Effect Upon Oenococcus oeni and Malolactic Fermentation

Directory of Open Access Journals (Sweden)

Aitor Balmaseda

2018-03-01

Full Text Available This work is a short review of the interactions between oenological yeasts and lactic acid bacteria (LAB, especially Oenococcus oeni, the main species carrying out the malolactic fermentation (MLF. The emphasis has been placed on non-Saccharomyces effects due to their recent increased interest in winemaking. Those interactions are variable, ranging from inhibitory, to neutral and stimulatory and are mediated by some known compounds, which will be discussed. One phenomena responsible of inhibitory interactions is the media exhaustion by yeasts, and particularly a decrease in L-malic acid by some non-Saccharomyces. Clearly ethanol is the main inhibitory compound of LAB produced by S. cerevisiae, but non-Saccharomyces can be used to decrease it. Sulfur dioxide and medium chain fatty acids (MCFAs produced by yeasts can exhibit inhibitory effect upon LAB or even result lethal. Interestingly mixed fermentations with non-Saccharomyces present less MCFA concentration. Among organic acids derived as result of yeast metabolism, succinic acid seems to be the most related with MLF inhibition. Several protein factors produced by S. cerevisiae inhibiting O. oeni have been described, but they have not been studied in non-Saccharomyces. According to the stimulatory effects, the use of non-Saccharomyces can increase the concentration of favorable mediators such as citric acid, pyruvic acid, or other compounds derived of yeast autolysis such as peptides, glucans, or mannoproteins. The emergence of non-Saccharomyces in winemaking present a new scenario in which MLF has to take place. For this reason, new tools and approaches should be explored to better understand this new winemaking context.

Non-Saccharomyces in Wine: Effect Upon Oenococcus oeni and Malolactic Fermentation.

Science.gov (United States)

Balmaseda, Aitor; Bordons, Albert; Reguant, Cristina; Bautista-Gallego, Joaquín

2018-01-01

This work is a short review of the interactions between oenological yeasts and lactic acid bacteria (LAB), especially Oenococcus oeni , the main species carrying out the malolactic fermentation (MLF). The emphasis has been placed on non- Saccharomyces effects due to their recent increased interest in winemaking. Those interactions are variable, ranging from inhibitory, to neutral and stimulatory and are mediated by some known compounds, which will be discussed. One phenomena responsible of inhibitory interactions is the media exhaustion by yeasts, and particularly a decrease in L-malic acid by some non- Saccharomyces . Clearly ethanol is the main inhibitory compound of LAB produced by S. cerevisiae , but non- Saccharomyces can be used to decrease it. Sulfur dioxide and medium chain fatty acids (MCFAs) produced by yeasts can exhibit inhibitory effect upon LAB or even result lethal. Interestingly mixed fermentations with non- Saccharomyces present less MCFA concentration. Among organic acids derived as result of yeast metabolism, succinic acid seems to be the most related with MLF inhibition. Several protein factors produced by S. cerevisiae inhibiting O. oeni have been described, but they have not been studied in non- Saccharomyces . According to the stimulatory effects, the use of non- Saccharomyces can increase the concentration of favorable mediators such as citric acid, pyruvic acid, or other compounds derived of yeast autolysis such as peptides, glucans, or mannoproteins. The emergence of non- Saccharomyces in winemaking present a new scenario in which MLF has to take place. For this reason, new tools and approaches should be explored to better understand this new winemaking context.

Functional Diversity of Haloacid Dehalogenase Superfamily Phosphatases from Saccharomyces cerevisiae: BIOCHEMICAL, STRUCTURAL, AND EVOLUTIONARY INSIGHTS.

Science.gov (United States)

Kuznetsova, Ekaterina; Nocek, Boguslaw; Brown, Greg; Makarova, Kira S; Flick, Robert; Wolf, Yuri I; Khusnutdinova, Anna; Evdokimova, Elena; Jin, Ke; Tan, Kemin; Hanson, Andrew D; Hasnain, Ghulam; Zallot, Rémi; de Crécy-Lagard, Valérie; Babu, Mohan; Savchenko, Alexei; Joachimiak, Andrzej; Edwards, Aled M; Koonin, Eugene V; Yakunin, Alexander F

2015-07-24

The haloacid dehalogenase (HAD)-like enzymes comprise a large superfamily of phosphohydrolases present in all organisms. The Saccharomyces cerevisiae genome encodes at least 19 soluble HADs, including 10 uncharacterized proteins. Here, we biochemically characterized 13 yeast phosphatases from the HAD superfamily, which includes both specific and promiscuous enzymes active against various phosphorylated metabolites and peptides with several HADs implicated in detoxification of phosphorylated compounds and pseudouridine. The crystal structures of four yeast HADs provided insight into their active sites, whereas the structure of the YKR070W dimer in complex with substrate revealed a composite substrate-binding site. Although the S. cerevisiae and Escherichia coli HADs share low sequence similarities, the comparison of their substrate profiles revealed seven phosphatases with common preferred substrates. The cluster of secondary substrates supporting significant activity of both S. cerevisiae and E. coli HADs includes 28 common metabolites that appear to represent the pool of potential activities for the evolution of novel HAD phosphatases. Evolution of novel substrate specificities of HAD phosphatases shows no strict correlation with sequence divergence. Thus, evolution of the HAD superfamily combines the conservation of the overall substrate pool and the substrate profiles of some enzymes with remarkable biochemical and structural flexibility of other superfamily members. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

KoVariome: Korean National Standard Reference Variome database of whole genomes with comprehensive SNV, indel, CNV, and SV analyses.

Science.gov (United States)

Kim, Jungeun; Weber, Jessica A; Jho, Sungwoong; Jang, Jinho; Jun, JeHoon; Cho, Yun Sung; Kim, Hak-Min; Kim, Hyunho; Kim, Yumi; Chung, OkSung; Kim, Chang Geun; Lee, HyeJin; Kim, Byung Chul; Han, Kyudong; Koh, InSong; Chae, Kyun Shik; Lee, Semin; Edwards, Jeremy S; Bhak, Jong

2018-04-04

High-coverage whole-genome sequencing data of a single ethnicity can provide a useful catalogue of population-specific genetic variations, and provides a critical resource that can be used to more accurately identify pathogenic genetic variants. We report a comprehensive analysis of the Korean population, and present the Korean National Standard Reference Variome (KoVariome). As a part of the Korean Personal Genome Project (KPGP), we constructed the KoVariome database using 5.5 terabases of whole genome sequence data from 50 healthy Korean individuals in order to characterize the benign ethnicity-relevant genetic variation present in the Korean population. In total, KoVariome includes 12.7M single-nucleotide variants (SNVs), 1.7M short insertions and deletions (indels), 4K structural variations (SVs), and 3.6K copy number variations (CNVs). Among them, 2.4M (19%) SNVs and 0.4M (24%) indels were identified as novel. We also discovered selective enrichment of 3.8M SNVs and 0.5M indels in Korean individuals, which were used to filter out 1,271 coding-SNVs not originally removed from the 1,000 Genomes Project when prioritizing disease-causing variants. KoVariome health records were used to identify novel disease-causing variants in the Korean population, demonstrating the value of high-quality ethnic variation databases for the accurate interpretation of individual genomes and the precise characterization of genetic variations.

The propagation of Escherichia Coli and of conservative tracers. A comparison

International Nuclear Information System (INIS)

Alexander, I.; Seiler, K.P.

1982-01-01

The propagation of Escherichia Coli (ATCC 11229, Gelsenkirchen) is compared with that of conservative tracers in groundwater. The experiments were performed with injection quantities of 10 7 , 10 8 , 10 10 and 10 11 of Escherichia Coli. Both, bacteria and conservative tracers pass their maximum at the same instant in the observation gauges. With injection quantities of more than 10 8 , the propagation of the Escherichia Coli sets in at the same time as it begins with the dyes. When the quantities range below 10 8 , the propagation begins after that of conservative tracers, because Coli bacteria were measured with a lower degree of detecting sensitivity than the tracers. With Coli injection quantities ranging above 10 10 , an increased filtering of these bacteria can be observed. Coli bacteria propagate more laterally than conservative tracers, however it could not be proved that this lateral propagation depends on the bacteria concentration. (orig.) [de

Molecular characterization and phylogeny of Shiga toxin-producing E. coli (STEC) from imported beef meat in Malaysia.

Science.gov (United States)

Abuelhassan, Nawal Nouridaim; Mutalib, Sahilah Abdul; Gimba, Fufa Ido; Yusoff, Wan Mohtar

2016-09-01

This study aimed at determining the presence and characterization of Escherichia coli and Shiga toxin-producing E. coli (STEC) from imported frozen beef meats. Seventy-four (74) frozen imported beef meat samples from two countries, India (42 samples) and Australia (32 samples), were collected and tested for E. coli. These samples were purchased from the frozen meat sections of five different supermarkets in different locations in Selangor, Malaysia, from April 2012 to October 2014. A total of 222 E. coli strains were isolated from the meat samples; 126 strains were isolated from country A (India), and 96 E. coli strains were from country of origin B (Australia), respectively. A total of 70 E. coli strains were identified and characterized. All E. coli strains were isolated into Fluorocult medium and identified using API 20E kit. All selected E. coli strains were characterized for Shiga toxin genes (stx1 and stx2). All biochemically identified E. coli in this study were further subjected to molecular detection through polymerase chain reaction (PCR) amplification and characterization using 16S ribosomal RNA (rRNA) gene of Shiga toxin-producing E. coli. Of the 70 E. coli strains, 11 strains were positive for both Shiga toxin genes (stx1 and stx2) and 11 (11/70) strains were positive for stx1 gene, while 25 (25/70) strains were positive for stx2 gene. The analysis of 16S rRNA gene of all the E. coli isolates in this study was successfully sequenced and analyzed, and based on sequence data obtained, a phylogenetic tree of the 16S rRNA gene was performed using Clustal W programme in MEGA 6.06 software. Phylogenetic tree showed that the E. coli isolates in our study cluster with the strain of E. coli isolated in other countries, which further confirm that the isolates of E. coli in this study are similar to those obtained in other studies. As a result, all the strains obtained in this study proved to be a strain of pathogenic E. coli, which may cause a serious outbreak

Functional expression of rat VPAC1 receptor in Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Hansen, M.K.; Tams, J.W.; Fahrenkrug, Jan

1999-01-01

G protein-coupled receptor; heterologous expression; membrane protein; Saccharomyces cerevisiae, vasoactive intestinal polypeptide; yeast mating factor-pre-pro *Ga-leader peptide......G protein-coupled receptor; heterologous expression; membrane protein; Saccharomyces cerevisiae, vasoactive intestinal polypeptide; yeast mating factor-pre-pro *Ga-leader peptide...

Both chronic treatments by epothilone D and fluoxetine increase the short-term memory and differentially alter the mood status of STOP/MAP6 KO mice.

Science.gov (United States)

Fournet, Vincent; de Lavilléon, Gaetan; Schweitzer, Annie; Giros, Bruno; Andrieux, Annie; Martres, Marie-Pascale

2012-12-01

Recent evidence underlines the crucial role of neuronal cytoskeleton in the pathophysiology of psychiatric diseases. In this line, the deletion of STOP/MAP6 (Stable Tubule Only Polypeptide), a microtubule-stabilizing protein, triggers various neurotransmission and behavioral defects, suggesting that STOP knockout (KO) mice could be a relevant experimental model for schizoaffective symptoms. To establish the predictive validity of such a mouse line, in which the brain serotonergic tone is dramatically imbalanced, the effects of a chronic fluoxetine treatment on the mood status of STOP KO mice were characterized. Moreover, we determined the impact, on mood, of a chronic treatment by epothilone D, a taxol-like microtubule-stabilizing compound that has previously been shown to improve the synaptic plasticity deficits of STOP KO mice. We demonstrated that chronic fluoxetine was either antidepressive and anxiolytic, or pro-depressive and anxiogenic, depending on the paradigm used to test treated mutant mice. Furthermore, control-treated STOP KO mice exhibited paradoxical behaviors, compared with their clear-cut basal mood status. Paradoxical fluoxetine effects and control-treated STOP KO behaviors could be because of their hyper-reactivity to acute and chronic stress. Interestingly, both epothilone D and fluoxetine chronic treatments improved the short-term memory of STOP KO mice. Such treatments did not affect the serotonin and norepinephrine transporter densities in cerebral areas of mice. Altogether, these data demonstrated that STOP KO mice could represent a useful model to study the relationship between cytoskeleton, mood, and stress, and to test innovative mood treatments, such as microtubule-stabilizing compounds. © 2012 The Authors Journal of Neurochemistry © 2012 International Society for Neurochemistry.

Provenience studies of the Ko-Kutani ware by neutron activation analysis

International Nuclear Information System (INIS)

Kawashima, Tatsuro.

1990-01-01

The instrumental neutron activation analysis has been applied to the multielemental analysis of eight samples of the Ko-Kutani ware including the Aote's that is one of semi-porcelain body. Each sample was compared with concentration of the useful index element, such as Ta, Sc, Ba, Hf, Th and rare earth, to discriminate the Arita group from the Kutani group. Eight samples were judged as products of the Arita, just as in previous paper. (author)

Identification and functional analysis of the Saccharomyces cerevisiae nicotinamidase gene, PNC1.

Science.gov (United States)

Ghislain, Michel; Talla, Emmanuel; François, Jean M

2002-02-01

Nicotinamidase (NAMase) from the budding yeast, Saccharomyces cerevisiae, was purified by Ni(2+) affinity chromatography and gel filtration. N-terminal microsequencing revealed sequence identity with a hypothetical polypeptide encoded by the yeast YGL037C open reading frame sharing 30% sequence identity with Escherichia coli pyrazinamidase/nicotinamidase. A yeast strain in which the NAMase gene, hereafter named PNC1, was deleted shows a decreased intracellular NAD(+) concentration, consistent with the loss of NAMase activity in the null mutant. In wild-type strains, NAMase activity is stimulated during the stationary phase of growth, by various hyperosmotic shocks or by ethanol treatment. Using a P(PNC1)::lacZ gene fusion, we have shown that this stimulation of NAMase activity results from increased levels of the protein and requires stress response elements in the 5'non-coding region of PNC1. These results suggest that NAMase helps yeast cells to adapt to various stress conditions and nutrient depletion, most likely via the activation of NAD-dependent biological processes. Copyright 2002 John Wiley & Sons, Ltd.

Review of Saccharomyces boulardii as a treatment option in IBD.

Science.gov (United States)

Sivananthan, Kavitha; Petersen, Andreas Munk

2018-05-17

Review of the yeast Saccharomyces boulardii as a treatment option for the inflammatory bowel diseases (IBD) ulcerative colitis and Crohn's disease. IBD is caused by an inappropriate immune response to gut microbiota. Treatment options could therefore be prebiotics, probiotics, antibiotics and/or fecal transplant. In this review, we have looked at the evidence for the yeast S. boulardii as a treatment option. Searches in PubMed and the Cochrane Library with the MeSH words 'Saccharomyces boulardii AND IBD', 'Saccharomyces boulardii AND Inflammatory Bowel Disease', 'Saccharomyces boulardii AND ulcerative colitis' and 'Saccharomyces boulardii AND Crohn's disease' gave total a total of 80 articles. After exclusions because of irrelevance, articles in other languages and some articles that were not available, 16 articles were included in this review. Three of the clinical trials showed a positive effect of S. boulardii in IBD patients (two Crohn's disease, one ulcerative colitis), while there was one trial that didn't prove any effect (Crohn's disease). Included Animal trials and cell assays describes different anti-inflammatory mechanisms of S. boulardii supporting a possible effect when treating IBD patients. The number of studies of S. boulardii as treatment for IBD is limited. Furthermore, the existing trials have small populations and short duration. We do not have enough evidence to prove the effect of S. boulardii in IBD. Saccharomyces boulardii is, however, a plausible treatment option in the future, but more placebo-controlled clinical studies on both patients with ulcerative colitis and Crohn's disease are needed.

Phosphorylation of Dgk1 Diacylglycerol Kinase by Casein Kinase II Regulates Phosphatidic Acid Production in Saccharomyces cerevisiae*

Science.gov (United States)

Qiu, Yixuan; Hassaninasab, Azam; Han, Gil-Soo; Carman, George M.

2016-01-01

In the yeast Saccharomyces cerevisiae, Dgk1 diacylglycerol (DAG) kinase catalyzes the CTP-dependent phosphorylation of DAG to form phosphatidic acid (PA). The enzyme in conjunction with Pah1 PA phosphatase controls the levels of PA and DAG for the synthesis of triacylglycerol and membrane phospholipids, the growth of the nuclear/endoplasmic reticulum membrane, and the formation of lipid droplets. Little is known about how DAG kinase activity is regulated by posttranslational modification. In this work, we examined the phosphorylation of Dgk1 DAG kinase by casein kinase II (CKII). When phosphate groups were globally reduced using nonspecific alkaline phosphatase, Triton X-100-solubilized membranes from DGK1-overexpressing cells showed a 7.7-fold reduction in DAG kinase activity; the reduced enzyme activity could be increased 5.5-fold by treatment with CKII. Dgk1(1–77) expressed heterologously in Escherichia coli was phosphorylated by CKII on a serine residue, and its phosphorylation was dependent on time as well as on the concentrations of CKII, ATP, and Dgk1(1–77). We used site-specific mutagenesis, coupled with phosphorylation analysis and phosphopeptide mapping, to identify Ser-45 and Ser-46 of Dgk1 as the CKII target sites, with Ser-46 being the major phosphorylation site. In vivo, the S46A and S45A/S46A mutations of Dgk1 abolished the stationary phase-dependent stimulation of DAG kinase activity. In addition, the phosphorylation-deficient mutations decreased Dgk1 function in PA production and in eliciting pah1Δ phenotypes, such as the expansion of the nuclear/endoplasmic reticulum membrane, reduced lipid droplet formation, and temperature sensitivity. This work demonstrates that the CKII-mediated phosphorylation of Dgk1 regulates its function in the production of PA. PMID:27834677

Distribution patterns of Saccharomyces species in cultural landscapes of Germany.

Science.gov (United States)

Brysch-Herzberg, Michael; Seidel, Martin

2017-08-01

The distribution patterns of the three Saccharomyces species, Saccharomyces paradoxus, S. uvarum and S. cerevisiae, were investigated by a culture-dependent approach in order to understand better how these species propagate in the cultural landscape of Germany. Saccharomyces paradoxus, the closest relative of S. cerevisiae, is shown to be a true woodland species. It was frequently found in the soil under conifers indicating that S. paradoxus is an autochthonous member of the microbial community in this habitat. Physiological characteristics of the species like the Crabtree effect and high tolerance against ethanol suggest that the species is adapted to regular supply with considerable amounts of sugars. Additionally, a high proportion of the S. paradoxus strains isolated in this study are shown to have the rare ability to ferment melezitose. For these reasons, it is hypothesized that S. paradoxus may be closely associated with the honeydew system in forests. Saccharomyces cerevisiae was rare in most habitats and only exceeded the frequency of S. paradoxus in habitats characterized by modern agricultural mass production of fruit. Both the landscape structure and the agricultural system heavily influence the frequencies of Saccharomyces species. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Introducing a new breed of wine yeast: interspecific hybridisation between a commercial Saccharomyces cerevisiae wine yeast and Saccharomyces mikatae.

Science.gov (United States)

Bellon, Jennifer R; Schmid, Frank; Capone, Dimitra L; Dunn, Barbara L; Chambers, Paul J

2013-01-01

Interspecific hybrids are commonplace in agriculture and horticulture; bread wheat and grapefruit are but two examples. The benefits derived from interspecific hybridisation include the potential of generating advantageous transgressive phenotypes. This paper describes the generation of a new breed of wine yeast by interspecific hybridisation between a commercial Saccharomyces cerevisiae wine yeast strain and Saccharomyces mikatae, a species hitherto not associated with industrial fermentation environs. While commercially available wine yeast strains provide consistent and reliable fermentations, wines produced using single inocula are thought to lack the sensory complexity and rounded palate structure obtained from spontaneous fermentations. In contrast, interspecific yeast hybrids have the potential to deliver increased complexity to wine sensory properties and alternative wine styles through the formation of novel, and wider ranging, yeast volatile fermentation metabolite profiles, whilst maintaining the robustness of the wine yeast parent. Screening of newly generated hybrids from a cross between a S. cerevisiae wine yeast and S. mikatae (closely-related but ecologically distant members of the Saccharomyces sensu stricto clade), has identified progeny with robust fermentation properties and winemaking potential. Chemical analysis showed that, relative to the S. cerevisiae wine yeast parent, hybrids produced wines with different concentrations of volatile metabolites that are known to contribute to wine flavour and aroma, including flavour compounds associated with non-Saccharomyces species. The new S. cerevisiae x S. mikatae hybrids have the potential to produce complex wines akin to products of spontaneous fermentation while giving winemakers the safeguard of an inoculated ferment.

Optimization of carbon and nitrogen medium components for biomass production using non-Saccharomyces wine yeasts.

Science.gov (United States)

Schnierda, T; Bauer, F F; Divol, B; van Rensburg, E; Görgens, J F

2014-05-01

The impact of different nitrogen and carbon sources on biomass production of the non-Saccharomyces wine yeast species Lachancea thermotolerans, Metschnikowia pulcherrima and Issatchenkia orientalis was assessed. Using a molasses-based medium, yeast extract and corn steep liquor as well as ammonium sulphate and di-ammonium phosphate (DAP) as nitrogen sources were compared in shake-flask cultures. A medium with 20 g l⁻¹ sugar (diluted molasses) and 500 mg l⁻¹ total yeast assimilable nitrogen, from yeast extract, gave the highest biomass concentrations and yields. Invertase pretreatment was required for cultures of M. pulcherrima and I. orientalis, and respective biomass yields of 0.7 and 0.8 g g⁻¹ were achieved in aerobic bioreactor cultures. The absence of ethanol production suggested Crabtree-negative behaviour by these yeasts, whereas Crabtree-positive behaviour by L. thermotolerans resulted in ethanol and biomass concentrations of 5.5 and 11.1 g l⁻¹, respectively. Recent studies demonstrate that non-Saccharomyces yeasts confer positive attributes to the final composition of wine. However, optimal process conditions for their biomass production have not been described, thereby limiting commercial application. In this study, industrial media and methods of yeast cultivation were investigated to develop protocols for biomass production of non-Saccharomyces yeast starter cultures for the wine industry. © 2014 The Society for Applied Microbiology.

Characterization of ent-kaurene synthase and kaurene oxidase involved in gibberellin biosynthesis from Scoparia dulcis.

Science.gov (United States)

Yamamura, Yoshimi; Taguchi, Yukari; Ichitani, Kei; Umebara, Io; Ohshita, Ayako; Kurosaki, Fumiya; Lee, Jung-Bum

2018-03-01

Gibberellins (GAs) are ubiquitous diterpenoids in higher plants, whereas some higher plants produce unique species-specific diterpenoids. In GA biosynthesis, ent-kaurene synthase (KS) and ent-kaurene oxidase (KO) are key players which catalyze early step(s) of the cyclization and oxidation reactions. We have studied the functional characterization of gene products of a KS (SdKS) and two KOs (SdKO1 and SdKO2) involved in GA biosynthesis in Scoparia dulcis. Using an in vivo heterologous expression system of Escherichia coli, we found that SdKS catalyzed a cyclization reaction from ent-CPP to ent-kaurene and that the SdKOs oxidized ent-kaurene to ent-kaurenoic acid after modification of the N-terminal region for adaptation to the E. coli expression system. The real-time PCR results showed that the SdKS, SdKO1 and SdKO2 genes were mainly expressed in the root and lateral root systems, which are elongating tissues. Based on these results, we suggest that these three genes may be responsible for the metabolism of GAs in S. dulcis.

Genetic diversity analysis of Amomum tsao-ko in Jinping County of Yunnan Province using SSR markers

Science.gov (United States)

Ma, Mengli; Wang, Tiantao; Lei, En; Meng, Hengling; Xie, Linyan; Zhu, Kunlong; Duan, Shaoze; Li, Wenqiang; Lu, Bingyue

2017-08-01

Genetic diversity analysis is very important for germplasm resources conservation and utilization. The objective of this study was to assess the genetic diversity among 44 individuals of Amomum tsao-ko from Jinping County of Yunnan Province using 5 selected SSR (simple sequence repeat) markers. A total of 23 polymorphic loci were detected among these germplasms, with an average of 4.6 polymorphic loci per SSR primer combination. The percentage of polymorphic loci was 100%, whereas the mean effective number of alleles (Ne), observed heterozygosity(Ho), expected heterozygosity (He), Shannon's information index (I), and the mean polymorphism information content (PIC) were 3. 410, 0. 491, 0. 679, 1.266 and 0. 672, respectively, indicating that the Amomum tsao-ko germplasms from Jinping County had high genetic diversity.

Beta3 adrenoceptors substitute the role of M(2) muscarinic receptor in coping with cold stress in the heart: evidence from M(2)KO mice.

Science.gov (United States)

Benes, Jan; Novakova, Martina; Rotkova, Jana; Farar, Vladimir; Kvetnansky, Richard; Riljak, Vladimir; Myslivecek, Jaromir

2012-07-01

We investigated the role of beta3-adrenoceptors (AR) in cold stress (1 or 7 days in cold) in animals lacking main cardioinhibitive receptors-M2 muscarinic receptors (M(2)KO). There was no change in receptor number in the right ventricles. In the left ventricles, there was decrease in binding to all cardiostimulative receptors (beta1-, and beta2-AR) and increase in cardiodepressive receptors (beta3-AR) in unstressed KO in comparison to WT. The cold stress in WT animals resulted in decrease in binding to beta1- and beta2-AR (to 37%/35% after 1 day in cold and to 27%/28% after 7 days in cold) while beta3-AR were increased (to 216% of control) when 7 days cold was applied. MR were reduced to 46% and 58%, respectively. Gene expression of M2 MR in WT was not changed due to stress, while M3 was changed. The reaction of beta1- and beta2-AR (binding) to cold was similar in KO and WT animals, and beta3-AR in stressed KO animals did not change. Adenylyl cyclase activity was affected by beta3-agonist CL316243 in cold stressed WT animals but CL316243 had almost no effects on adenylyl cyclase activity in stressed KO. Nitric oxide activity (NOS) was not affected by BRL37344 (beta3-agonist) both in WT and KO animals. Similarly, the stress had no effects on NOS activity in WT animals and in KO animals. We conclude that the function of M2 MR is substituted by beta3-AR and that these effects are mediated via adenylyl cyclase rather than NOS.

Yeast redoxyendonuclease, a DNA repair enzyme similar to Escherichia coli endonuclease III

International Nuclear Information System (INIS)

Gossett, J.; Lee, K.; Cunningham, R.P.; Doetsch, P.W.

1988-01-01

A DNA repair endonuclease (redoxyendonuclease) was isolated from bakers' yeast (Saccharomyces cerevisiae). The enzyme has been purified by a series of column chromatography steps and cleaves OsO 4 -damaged, double-stranded DNA at sites of thymine glycol and heavily UV-irradiated DNA at sites of cytosine, thymine, and guanine photoproducts. The base specificity and mechanism of phosphodiester bond cleavage for the yeast redoxyendonuclease appear to be identical with those of Escherichia coli endonuclease III when thymine glycol containing, end-labeled DNA fragments of defined sequence are employed as substrates. Yeast redoxyendonuclease has an apparent molecular size of 38,000-42,000 daltons and is active in the absence of divalent metal cations. The identification of such an enzyme in yeast may be of value in the elucidation of the biochemical basis for radiation sensitivity in certain yeast mutants

Heterogeneity in Induction Level, Infection Ability, and Morphology of Shiga Toxin-Encoding Phages (Stx Phages) from Dairy and Human Shiga Toxin-Producing Escherichia coli O26:H11 Isolates

Science.gov (United States)

Bonanno, Ludivine; Petit, Marie-Agnès; Loukiadis, Estelle; Michel, Valérie

2016-01-01

Shiga toxin (Stx)-producing Escherichia coli (STEC) bacteria are foodborne pathogens responsible for diarrhea and hemolytic-uremic syndrome (HUS). Shiga toxin, the main STEC virulence factor, is encoded by the stx gene located in the genome of a bacteriophage inserted into the bacterial chromosome. The O26:H11 serotype is considered to be the second-most-significant HUS-causing serotype worldwide after O157:H7. STEC O26:H11 bacteria and their stx-negative counterparts have been detected in dairy products. They may convert from the one form to the other by loss or acquisition of Stx phages, potentially confounding food microbiological diagnostic methods based on stx gene detection. Here we investigated the diversity and mobility of Stx phages from human and dairy STEC O26:H11 strains. Evaluation of their rate of in vitro induction, occurring either spontaneously or in the presence of mitomycin C, showed that the Stx2 phages were more inducible overall than Stx1 phages. However, no correlation was found between the Stx phage levels produced and the origin of the strains tested or the phage insertion sites. Morphological analysis by electron microscopy showed that Stx phages from STEC O26:H11 displayed various shapes that were unrelated to Stx1 or Stx2 types. Finally, the levels of sensitivity of stx-negative E. coli O26:H11 to six Stx phages differed among the 17 strains tested and our attempts to convert them into STEC were unsuccessful, indicating that their lysogenization was a rare event. PMID:26826235

Influence of organic acids and organochlorinated insecticides on metabolism of Saccharomyces cerevisiae

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Pejin Dušanka J.

2005-01-01

Full Text Available Saccharomyces cerevisiae is exposed to different stress factors during the production: osmotic, temperature, oxidative. The response to these stresses is the adaptive mechanism of cells. The raw materials Saccharomyces cerevisiae is produced from, contain metabolism products of present microorganisms and protective agents used during the growth of sugar beet for example the influence of acetic and butyric acid and organochlorinated insecticides, lindan and heptachlor, on the metabolism of Saccharomyces cerevisiae was investigated and presented in this work. The mentioned compounds affect negatively the specific growth rate, yield, content of proteins, phosphorus, total ribonucleic acids. These compounds influence the increase of trechalose and glycogen content in the Saccharomyces cerevisiae cells.

Evidence for Divergent Evolution of Growth Temperature Preference in Sympatric Saccharomyces Species

Science.gov (United States)

Gonçalves, Paula; Valério, Elisabete; Correia, Cláudia; de Almeida, João M. G. C. F.; Sampaio, José Paulo

2011-01-01

The genus Saccharomyces currently includes eight species in addition to the model yeast Saccharomyces cerevisiae, most of which can be consistently isolated from tree bark and soil. We recently found sympatric pairs of Saccharomyces species, composed of one cryotolerant and one thermotolerant species in oak bark samples of various geographic origins. In order to contribute to explain the occurrence in sympatry of Saccharomyces species, we screened Saccharomyces genomic data for protein divergence that might be correlated to distinct growth temperature preferences of the species, using the dN/dS ratio as a measure of protein evolution rates and pair-wise species comparisons. In addition to proteins previously implicated in growth at suboptimal temperatures, we found that glycolytic enzymes were among the proteins exhibiting higher than expected divergence when one cryotolerant and one thermotolerant species are compared. By measuring glycolytic fluxes and glycolytic enzymatic activities in different species and at different temperatures, we subsequently show that the unusual divergence of glycolytic genes may be related to divergent evolution of the glycolytic pathway aligning its performance to the growth temperature profiles of the different species. In general, our results support the view that growth temperature preference is a trait that may have undergone divergent selection in the course of ecological speciation in Saccharomyces. PMID:21674061

Construction of killer industrial yeast Saccharomyces cerevisiae HAU-1 and its fermentation performance

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Bijender K. Bajaj

2010-06-01

Full Text Available Saccharomyces cerevisiae HAU-1, a time tested industrial yeast possesses most of the desirable fermentation characteristics like fast growth and fermentation rate, osmotolerance, high ethanol tolerance, ability to ferment molasses, and to ferment at elevated temperatures etc. However, this yeast was found to be sensitive against the killer strains of Saccharomyces cerevisiae. In the present study, killer trait was introduced into Saccharomyces cerevisiae HAU-1 by protoplast fusion with Saccharomyces cerevisiae MTCC 475, a killer strain. The resultant fusants were characterized for desirable fermentation characteristics. All the technologically important characteristics of distillery yeast Saccharomyces cerevisiae HAU-1 were retained in the fusants, and in addition the killer trait was also introduced into them. Further, the killer activity was found to be stably maintained during hostile conditions of ethanol fermentations in dextrose or molasses, and even during biomass recycling.

The effect of essential oil of basil (Ocimum basilicum L.) on UV-induced mutagenesis in Escherichia coli and Saccharomyces cerevisiae

Energy Technology Data Exchange (ETDEWEB)

Stanojević, Jasna; Berić, Tanja; Opačić, Biljana; Vuković-Gačić, Branka; Simić, Draga; Knežević-Vukčević, Jelena [Institute of Botany, Faculty of Biology, University of Belgrade, 11000 Belgrade (Serbia)

2008-07-01

The antimutagenic potential of essential oil (EO) of basil (Ocimum basilicum L.) and its major constituent linalool were studied with the E. coli K12 and S. cerevisiae D7 assays. In the E. coli assay, EO and linalool inhibited UV-induced mutagenesis in a repair-proficient strain, but had no effect on spontaneous mutagenesis in repair-proficient, nucleotide excision repair-deficient, and mismatch-deficient strains. By testing participation of different mechanisms involved in antimutagenesis, it was concluded that the antimutagenic effect against UV-induced mutagenesis involved decrease of protein synthesis and cell proliferation which led to increased efficiency of nucleotide excision repair. An antimutagenic effect of basil derivatives in S. cerevisiae was not detected. (author)

The effect of essential oil of basil (Ocimum basilicum L.) on UV-induced mutagenesis in Escherichia coli and Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Stanojević, Jasna; Berić, Tanja; Opačić, Biljana; Vuković-Gačić, Branka; Simić, Draga; Knežević-Vukčević, Jelena

2008-01-01

The antimutagenic potential of essential oil (EO) of basil (Ocimum basilicum L.) and its major constituent linalool were studied with the E. coli K12 and S. cerevisiae D7 assays. In the E. coli assay, EO and linalool inhibited UV-induced mutagenesis in a repair-proficient strain, but had no effect on spontaneous mutagenesis in repair-proficient, nucleotide excision repair-deficient, and mismatch-deficient strains. By testing participation of different mechanisms involved in antimutagenesis, it was concluded that the antimutagenic effect against UV-induced mutagenesis involved decrease of protein synthesis and cell proliferation which led to increased efficiency of nucleotide excision repair. An antimutagenic effect of basil derivatives in S. cerevisiae was not detected. (author)

Fungal genomics beyond Saccharomyces cerevisiae?

DEFF Research Database (Denmark)

Hofmann, Gerald; Mcintyre, Mhairi; Nielsen, Jens

2003-01-01

Fungi are used extensively in both fundamental research and industrial applications. Saccharomyces cerevisiae has been the model organism for fungal research for many years, particularly in functional genomics. However, considering the diversity within the fungal kingdom, it is obvious...

Contemporary population and development challenges in the Municipality of Dubrovačko Primorje, Croatia

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Helena Karlić Mujo

2010-06-01

Full Text Available The purpose of this paper is to define the basic issues and opportunities challenging the modern development of the Municipality of Dubrovačko Primorje, their causes and effects. The processes of depopulation affected this poorly developed rural region since 1960s, which had been influenced by various factors: development polarisation caused by intensive development of tourism, the poor traffic infrastructure in the highland-area of the Municipality and the unsolved water supply issue. In 1990s a series of events intensified negative demographic trends as follows: aggression on Croatia, occupation during which residential, economic and infrastructural buildings were destroyed and burnt, as well as an earthquake which happened after the return of the refugees. The long-lasting rural emigration from this region towards Dubrovnik caused not only an instantaneous decrease of population but also a contraction of the renewing basis and an accelerated decrease of natural growth in the Municipality of Dubrovačko Primorje. In that way the emigrational depopulation as a consequence of a specific socioeconomic development, became the cause of the current biological depopulation. The intensity of the depopulation processes has been established by an analysis of a total population dynamics and the changes in biological and economic population structure. The conclusion of the study indicates activities which have promising development in the Municipality of Dubrovačko Primorje and which might partly stop the negative demographic trends. Those are mainly traditional activities in this region which are not commercially used enough. They should prevent any further emigration and perhaps even attract those who immigrated, back to their place of birth.

Contemporary population and development challenges in the Municipality of Dubrovačko Primorje, Croatia

Directory of Open Access Journals (Sweden)

Helena Karlić Mujo

2010-01-01

Full Text Available The purpose of this paper is to define the basic issues and opportunities challenging the modern development of the Municipality of Dubrovačko Primorje, their causes and effects. The processes of depopulation affected this poorly developed rural region since 1960s, which had been influenced by various factors: development polarisation caused by intensive development of tourism, the poor traffic infrastructure in the highland-area of the Municipality and the unsolved water supply issue. In 1990s a series of events intensified negative demographic trends as follows: aggression on Croatia, occupation during which residential, economic and infrastructural buildings were destroyed and burnt, as well as an earthquake which happened after the return of the refugees. The long-lasting rural emigration from this region towards Dubrovnik caused not only an instantaneous decrease of population but also a contraction of the renewing basis and an accelerated decrease of natural growth in the Municipality of Dubrovačko Primorje. In that way the emigrational depopulation as a consequence of a specific socioeconomic development, became the cause of the current biological depopulation. The intensity of the depopulation processes has been established by an analysis of a total population dynamics and the changes in biological and economic population structure. The conclusion of the study indicates activities which have promising development in the Municipality of Dubrovačko Primorje and which might partly stop the negative demographic trends. Those are mainly traditional activities in this region which are not commercially used enough. They should prevent any further emigration and perhaps even attract those who immigrated, back to their place of birth.

Introducing a new breed of wine yeast: interspecific hybridisation between a commercial Saccharomyces cerevisiae wine yeast and Saccharomyces mikatae.

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Jennifer R Bellon

Full Text Available Interspecific hybrids are commonplace in agriculture and horticulture; bread wheat and grapefruit are but two examples. The benefits derived from interspecific hybridisation include the potential of generating advantageous transgressive phenotypes. This paper describes the generation of a new breed of wine yeast by interspecific hybridisation between a commercial Saccharomyces cerevisiae wine yeast strain and Saccharomyces mikatae, a species hitherto not associated with industrial fermentation environs. While commercially available wine yeast strains provide consistent and reliable fermentations, wines produced using single inocula are thought to lack the sensory complexity and rounded palate structure obtained from spontaneous fermentations. In contrast, interspecific yeast hybrids have the potential to deliver increased complexity to wine sensory properties and alternative wine styles through the formation of novel, and wider ranging, yeast volatile fermentation metabolite profiles, whilst maintaining the robustness of the wine yeast parent. Screening of newly generated hybrids from a cross between a S. cerevisiae wine yeast and S. mikatae (closely-related but ecologically distant members of the Saccharomyces sensu stricto clade, has identified progeny with robust fermentation properties and winemaking potential. Chemical analysis showed that, relative to the S. cerevisiae wine yeast parent, hybrids produced wines with different concentrations of volatile metabolites that are known to contribute to wine flavour and aroma, including flavour compounds associated with non-Saccharomyces species. The new S. cerevisiae x S. mikatae hybrids have the potential to produce complex wines akin to products of spontaneous fermentation while giving winemakers the safeguard of an inoculated ferment.

Introducing a New Breed of Wine Yeast: Interspecific Hybridisation between a Commercial Saccharomyces cerevisiae Wine Yeast and Saccharomyces mikatae

Science.gov (United States)

Bellon, Jennifer R.; Schmid, Frank; Capone, Dimitra L.; Dunn, Barbara L.; Chambers, Paul J.

2013-01-01

Interspecific hybrids are commonplace in agriculture and horticulture; bread wheat and grapefruit are but two examples. The benefits derived from interspecific hybridisation include the potential of generating advantageous transgressive phenotypes. This paper describes the generation of a new breed of wine yeast by interspecific hybridisation between a commercial Saccharomyces cerevisiae wine yeast strain and Saccharomyces mikatae, a species hitherto not associated with industrial fermentation environs. While commercially available wine yeast strains provide consistent and reliable fermentations, wines produced using single inocula are thought to lack the sensory complexity and rounded palate structure obtained from spontaneous fermentations. In contrast, interspecific yeast hybrids have the potential to deliver increased complexity to wine sensory properties and alternative wine styles through the formation of novel, and wider ranging, yeast volatile fermentation metabolite profiles, whilst maintaining the robustness of the wine yeast parent. Screening of newly generated hybrids from a cross between a S. cerevisiae wine yeast and S. mikatae (closely-related but ecologically distant members of the Saccharomyces sensu stricto clade), has identified progeny with robust fermentation properties and winemaking potential. Chemical analysis showed that, relative to the S. cerevisiae wine yeast parent, hybrids produced wines with different concentrations of volatile metabolites that are known to contribute to wine flavour and aroma, including flavour compounds associated with non-Saccharomyces species. The new S. cerevisiae x S. mikatae hybrids have the potential to produce complex wines akin to products of spontaneous fermentation while giving winemakers the safeguard of an inoculated ferment. PMID:23614011

Murine Colitis is Mediated by Vimentin

OpenAIRE

Mor-Vaknin, Nirit; Legendre, Maureen; Yu, Yue; Serezani, Carlos H. C.; Garg, Sanjay K.; Jatzek, Anna; Swanson, Michael D.; Gonzalez-Hernandez, Marta J.; Teitz-Tennenbaum, Seagal; Punturieri, Antonello; Engleberg, N. Cary; Banerjee, Ruma; Peters-Golden, Marc; Kao, John Y.; Markovitz, David M.

2013-01-01

Vimentin, an abundant intermediate filament protein, presumably has an important role in stabilizing intracellular architecture, but its function is otherwise poorly understood. In a vimentin knockout (Vim KO) mouse model, we note that Vim KO mice challenged with intraperitoneal Escherichia coli control bacterial infection better than do wild-type (WT) mice. In vitro, Vim KO phagocytes show significantly increased capacity to mediate bacterial killing by abundant production of reactive oxygen...

Pathogenic Escherichia coli and food handlers in luxury hotels in Nairobi, Kenya.

Science.gov (United States)

Onyango, Abel O; Kenya, Eucharia U; Mbithi, John J N; Ng'ayo, Musa O

2009-11-01

The epidemiology and virulence properties of pathogenic Escherichia coli among food handlers in tourist destination hotels in Kenya are largely uncharacterized. This cross-sectional study among consenting 885 food handlers working in nine luxurious tourist hotels in Nairobi, Kenya determined the epidemiology, virulence properties, antibiotics susceptibility profiles and conjugation abilities of pathogenic Escherichia coli. Pathogenic Escherichia coli was detected among 39 (4.4%) subjects, including 1.8% enteroaggregative Escherichia coli (EAEC) harboring aggR genes, 1.2% enterotoxigenic Escherichia coli (ETEC) expressing both LT and STp toxins, 1.1% enteropathogenic Escherichia coli (EPEC) and 0.2% Shiga-like Escherichia coli (EHEC) both harboring eaeA and stx2 genes respectively. All the pathotypes had increased surface hydrophobicity. Using multivariate analyses, food handlers with loose stools were more likely to be infected with pathogenic Escherichia coli. Majority 53.8% of the pathotypes were resistant to tetracycline with 40.2% being multi-drug resistant. About 85.7% pathotypes trans-conjugated with Escherichia coli K12 F(-) NA(r) LA. The carriage of multi-drug resistant, toxin expressing pathogenic Escherichia coli by this population is of public health concern because exposure to low doses can result in infection. Screening food handlers and implementing public awareness programs is recommended as an intervention to control transmission of enteric pathogens.

Functional characterization of a rice de novo DNA methyltransferase, OsDRM2, expressed in Escherichia coli and yeast

Energy Technology Data Exchange (ETDEWEB)

Pang, Jinsong, E-mail: pangjs542@nenu.edu.cn [Key Laboratory of Molecular Epigenetics of the Ministry of Education, Northeast Normal University, Changchun, Jilin 130024 (China); Dong, Mingyue; Li, Ning; Zhao, Yanli [Key Laboratory of Molecular Epigenetics of the Ministry of Education, Northeast Normal University, Changchun, Jilin 130024 (China); Liu, Bao, E-mail: baoliu@nenu.edu.cn [Key Laboratory of Molecular Epigenetics of the Ministry of Education, Northeast Normal University, Changchun, Jilin 130024 (China)

2013-03-01

Highlights: ► A rice de novo DNA methyltransferase OsDRM2 was cloned. ► In vitro methylation activity of OsDRM2 was characterized with Escherichia coli. ► Assays of OsDRM2 in vivo methylation were done with Saccharomyces cerevisiae. ► OsDRM2 methylation activity is not preferential to any type of cytosine context. ► The activity of OsDRM2 is independent of RdDM pathway. - Abstract: DNA methylation of cytosine nucleotides is an important epigenetic modification that occurs in most eukaryotic organisms and is established and maintained by various DNA methyltransferases together with their co-factors. There are two major categories of DNA methyltransferases: de novo and maintenance. Here, we report the isolation and functional characterization of a de novo methyltransferase, named OsDRM2, from rice (Oryza sativa L.). The full-length coding region of OsDRM2 was cloned and transformed into Escherichia coli and Saccharomyces cerevisiae. Both of these organisms expressed the OsDRM2 protein, which exhibited stochastic de novo methylation activity in vitro at CG, CHG, and CHH di- and tri-nucleotide patterns. Two lines of evidence demonstrated the de novo activity of OsDRM2: (1) a 5′-CCGG-3′ containing DNA fragment that had been pre-treated with OsDRM2 protein expressed in E. coli was protected from digestion by the CG-methylation-sensitive isoschizomer HpaII; (2) methylation-sensitive amplified polymorphism (MSAP) analysis of S. cerevisiae genomic DNA from transformants that had been introduced with OsDRM2 revealed CG and CHG methylation levels of 3.92–9.12%, and 2.88–6.93%, respectively, whereas the mock control S. cerevisiae DNA did not exhibit cytosine methylation. These results were further supported by bisulfite sequencing of the 18S rRNA and EAF5 genes of the transformed S. cerevisiae, which exhibited different DNA methylation patterns, which were observed in the genomic DNA. Our findings establish that OsDRM2 is an active de novo DNA

Functional characterization of a rice de novo DNA methyltransferase, OsDRM2, expressed in Escherichia coli and yeast

International Nuclear Information System (INIS)

Pang, Jinsong; Dong, Mingyue; Li, Ning; Zhao, Yanli; Liu, Bao

2013-01-01

Highlights: ► A rice de novo DNA methyltransferase OsDRM2 was cloned. ► In vitro methylation activity of OsDRM2 was characterized with Escherichia coli. ► Assays of OsDRM2 in vivo methylation were done with Saccharomyces cerevisiae. ► OsDRM2 methylation activity is not preferential to any type of cytosine context. ► The activity of OsDRM2 is independent of RdDM pathway. - Abstract: DNA methylation of cytosine nucleotides is an important epigenetic modification that occurs in most eukaryotic organisms and is established and maintained by various DNA methyltransferases together with their co-factors. There are two major categories of DNA methyltransferases: de novo and maintenance. Here, we report the isolation and functional characterization of a de novo methyltransferase, named OsDRM2, from rice (Oryza sativa L.). The full-length coding region of OsDRM2 was cloned and transformed into Escherichia coli and Saccharomyces cerevisiae. Both of these organisms expressed the OsDRM2 protein, which exhibited stochastic de novo methylation activity in vitro at CG, CHG, and CHH di- and tri-nucleotide patterns. Two lines of evidence demonstrated the de novo activity of OsDRM2: (1) a 5′-CCGG-3′ containing DNA fragment that had been pre-treated with OsDRM2 protein expressed in E. coli was protected from digestion by the CG-methylation-sensitive isoschizomer HpaII; (2) methylation-sensitive amplified polymorphism (MSAP) analysis of S. cerevisiae genomic DNA from transformants that had been introduced with OsDRM2 revealed CG and CHG methylation levels of 3.92–9.12%, and 2.88–6.93%, respectively, whereas the mock control S. cerevisiae DNA did not exhibit cytosine methylation. These results were further supported by bisulfite sequencing of the 18S rRNA and EAF5 genes of the transformed S. cerevisiae, which exhibited different DNA methylation patterns, which were observed in the genomic DNA. Our findings establish that OsDRM2 is an active de novo DNA

Anaerobic organic acid metabolism of Candida zemplinina in comparison with Saccharomyces wine yeasts.

Science.gov (United States)

Magyar, Ildikó; Nyitrai-Sárdy, Diána; Leskó, Annamária; Pomázi, Andrea; Kállay, Miklós

2014-05-16

Organic acid production under oxygen-limited conditions has been thoroughly studied in the Saccharomyces species, but practically never investigated in Candida zemplinina, which seems to be an acidogenic species under oxidative laboratory conditions. In this study, several strains of C. zemplinina were tested for organic acid metabolism, in comparison with Saccharomyces cerevisiae, Saccharomyces uvarum and Candida stellata, under fermentative conditions. Only C. stellata produced significantly higher acidity in simple minimal media (SM) with low sugar content and two different nitrogen sources (ammonia or glutamic acid) at low level. However, the acid profile differed largely between the Saccharomyces and Candida species and showed inverse types of N-dependence in some cases. Succinic acid production was strongly enhanced on glutamic acid in Saccharomyces species, but not in Candida species. 2-oxoglutarate production was strongly supported on ammonium nitrogen in Candida species, but remained low in Saccharomyces. Candida species, C. stellata in particular, produced more pyruvic acid regardless of N-sources. From the results, we concluded that the anaerobic organic acid metabolisms of C. zemplinina and C. stellata are different from each other and also from that of the Saccharomyces species. In the formation of succinic acid, the oxidative pathway from glutamic acid seems to play little or no role in C. zemplinina. The reductive branch of the TCA cycle, however, produces acidic intermediates (malic, fumaric, and succinic acid) in a level comparable with the production of the Saccharomyces species. An unidentified organic acid, which was produced on glutamic acid only by the Candida species, needs further investigation. Copyright © 2014 Elsevier B.V. All rights reserved.

Experience of Ko-Ri Unit 1 water chemistry

International Nuclear Information System (INIS)

Tae Il Lee

1983-01-01

The main focus is placed on operational experience in secondary system water chemistry (especially the steam generator) of the Ko-Ri nuclear power plant Unit 1, Republic of Korea, but primary side chemistry is also discussed. The major concern of secondary water chemistry in a PWR is that the condition of the steam generator be well maintained. Full flow deep bed condensate polishers have recently been installed and operation started in July 1982. Boric acid treatment of the steam generator was stopped and only the all volatile treatment method was used thereafter. A review of steam generator integrity, the chemistry control programme, secondary water quality, etc. is considered to be of great value regarding the operation of Unit 1 and future units now under startup testing or construction in the Republic of Korea. (author)

Identification of new genes required for meiotic recombination in Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Ajimura, M.; Lee, S.H.; Ogawa, H.

1993-01-01

Mutants defective in meiotic recombination were isolated from a disomic haploid strain of Saccharomyces cerevisiae by examining recombination within the leu2 and his4 heteroalleles located on chromosome III. The mutants were classified into two new complementation groups (MRE2 and MRE11) and eight previously identified groups, which include SPO11, HOP1, REC114, MRE4/MEK1 and genes in the RAD52 epistasis group. All of the mutants, in which the mutations in the new complementation groups are homozygous and diploid, can undergo premeiotic DNA synthesis and produce spores. The spores are, however, not viable. The mre2 and mre11 mutants produce viable spores in a spo13 background, in which meiosis I is bypassed, suggesting that these mutants are blocked at an early step in meiotic recombination. The mre2 mutant does not exhibit any unusual phenotype during mitosis and it is, thus, considered to have a mutation in a meiosis-specific gene. By contrast, the mre11 mutant is sensitive to damage to DNA by methyl methanesulfonate and exhibits a hyperrecombination phenotype in mitosis. Among six alleles of HOP1 that were isolated, an unusual pattern of intragenic complementation was observed

Melanosis coli in patients with colon cancer

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Dorota Biernacka-Wawrzonek

2016-12-01

Full Text Available Intoduction: Melanosis coli is a benign lesion affecting the mucosa of the large intestine. There is a relationship between the presence of melanosis and anthraquinone laxative use. Melanosis coli is also observed in patients with colon cancer, but there is doubt whether these two conditions are related. Aim : To analyze the correlation between melanosis and colon cancer. Material and methods: We analyzed retrospectively 436 patients undergoing colon cancer surgery. There were 246 women and 190 men. Patients were divided into three age groups: under 50 years, between 51 and 65 years, and over 66 years. We analyzed sections of the cancer and intestinal mucosa from the tumor’s proximal (2–5 cm and distal (8–10 cm zone. Results : Melanosis coli was present in 52 patients, which represents 11.9% of patients with colon cancer. More often it was present in women. The most common location of melanosis and colon cancer was the terminal part of the large intestine. In patients below 50 years of age in both sexes melanosis coli did not occur. In men, melanosis was more common in the age group over 66 years. Intensity of pigmentation was higher in the tumor’s distal zone. Conclusions : The incidence of melanosis coli increases with age, similar to that of colon cancer. Melanosis was not present inside tumors, in almost half of the cases it was not present in the proximal zone, and the degree of pigmentation increased in distal zone. The cause-effect relationship between melanosis coli and colon cancer remains uncertain.

Phase analysis of Košice meteorite: Preliminary results

Science.gov (United States)

Sitek, J.; Dekan, J.; Degmová, J.; Sedlačková, K.

2012-10-01

Meteorite fall was observed by the Košice town in Slovakia in February 2010 and it was classified as an ordinary chondrite H5. The samples were prepared in powder form scratched from the surface. Mossbauer spectra were measured at room temperature and liquid nitrogen temperature. Spectra consist of components related to iron-bearing phases with different content. Non-magnetic part was fitted with three quadrupole doublets. According to its parameters, we identified olivine, pyroxene, and traces of Fe3+ phases. Magnetic part consists of an iron-rich Fe-Ni alloy with hyperfine magnetic field similar to kamacite α-Fe(Ni,Co) and troilite. Main elements were also determined by X-ray fluorescence spectroscopy.

Air-liquid biofilm formation is dependent on ammonium depletion in a Saccharomyces cerevisiae flor strain.

Science.gov (United States)

Zara, Giacomo; Budroni, Marilena; Mannazzu, Ilaria; Zara, Severino

2011-12-01

Air-liquid biofilm formation appears to be an adaptive mechanism that promotes foraging of Saccharomyces cerevisiae flor strains in response to nutrient starvation. The FLO11 gene plays a central role in this phenotype as its expression allows yeast cells to rise to the liquid surface. Here, we investigated the role of ammonium depletion in air-liquid biofilm formation and FLO11 expression in a S. cerevisiae flor strain. The data obtained show that increasing ammonium concentrations from 0 to 450 m m reduce air-liquid biofilm in terms of biomass and velum formation and correlate with a reduction of FLO11 expression. Rapamycin inhibition of the TOR pathway and deletion of RAS2 gene significantly reduced biofilm formation and FLO11 expression. Taken together, these data suggest that ammonium depletion is a key factor in the induction of air-liquid biofilm formation and FLO11 expression in S. cerevisiae flor strains. Copyright © 2011 John Wiley & Sons, Ltd.

Effects of the rad52 gene on recombination in Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Prakash, S.; Prakash, L.; Burke, W.; Montelone, B.A.

1980-01-01

Effects of the rad 52 mutation in Saccharomyces cerevisiae on meiotic, γ-ray-induced, uv-induced and spontaneous mitotic recombination were studied. The rad52/rad52 diploids undergo premeiotic DNA synthesis; sporulation occurs but inviable spores are produced. Both intra and intergenic recombination during meiosis were examined in cells transferred from sporulation medium to vegetative medium at different time intervals. No intragenic recombination was observed at the his1-1/his1-315 and trp-5-2/trp5-48 heteroalleles. Gene-centromere recombination also was not observed in rad/52/rad52 diploids. No γ-ray- or uv-induced intragenic mitotic recombination is seen in rad52/rad52 diploids. The rate of spontaneous mitotic recombination is lowered five-fold at the his1-1/his1-315 and leu1-c/leu1-12 heteroalleles. Spontaneous reversion rates of both his1-1 and his1-315 were elevated 10 to 20 fold in rad52/rad52 diploids. The RAD52 gene function is required for spontaneous mitotic recombination, uv- and γ-ray-induced mitotic recombination and mitotic recombination

The antimicrobial peptide cathelicidin protects mice from Escherichia coli O157:H7-mediated disease.

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Milan Chromek

Full Text Available This study investigated the role of the antimicrobial peptide cathelicidin in Escherichia coli O157:H7 infection and subsequent renal damage. Mouse and human cathelicidin, CRAMP and LL-37, respectively, killed E. coli O157:H7 in vitro. Intestines from healthy wild-type (129/SvJ and cathelicidin-knock-out (Camp(-/- mice were investigated, showing that cathelicidin-deficient mice had a thinner colonic mucus layer compared with wild-type mice. Wild-type (n = 11 and cathelicidin-knock-out (n = 11 mice were inoculated with E. coli O157:H7. Cathelicidin-deficient animals exhibited higher fecal counts of E. coli O157:H7 and bacteria penetrated the mucus forming attaching-and-effacing lesions to a much higher extent than in wild-type animals. Cathelicidin knock-out mice developed symptoms (9/11 as well as anemia, thrombocytopenia and extensive renal tubular damage while all cathelicidin-producing mice remained asymptomatic with normal laboratory findings. When injected with Shiga toxin intraperitoneally, both murine strains developed the same degree of renal tubular damage and clinical disease indicating that differences in sensitivity to infection between the murine strains were related to the initial intestinal response. In conclusion, cathelicidin substantially influenced the antimicrobial barrier in the mouse colon mucosa. Cathelicidin deficiency lead to increased susceptibility to E. coli O157:H7 infection and subsequent renal damage. Administration of cathelicidin or stimulation of endogenous production may prove to be novel treatments for E. coli O157:H7-induced hemolytic uremic syndrome.

Adenovirus-mediated sphingomyelin synthase 2 increases atherosclerotic lesions in ApoE KO mice

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Zhao Yarui

2011-01-01

Full Text Available Abstract Background Sphingomyelin synthase 2 (SMS2 contributes to de novo sphingomyelin (SM biosynthesis. Its activity is related to SM levels in the plasma and the cell membrane. In this study, we investigated the possibility of a direct relationship between SMS and atherosclerosis. Methods The Adenovirus containing SMS2 gene was given into 10-week ApoE KO C57BL/6J mice by femoral intravenous injection. In the control group, the Adenovirus containing GFP was given. To confirm this model, we took both mRNA level examination (RT-PCR and protein level examination (SMS activity assay. Result We generated recombinant adenovirus vectors containing either human SMS2 cDNA (AdV-SMS2 or GFP cDNA (AdV-GFP. On day six after intravenous infusion of 2 × 1011 particle numbers into ten-week-old apoE KO mice, AdV-SMS2 treatment significantly increased liver SMS2 mRNA levels and SMS activity (by 2.7-fold, 2.3-fold, p Conclusions Our results present direct morphological evidence for the pro-atherogenic capabilities of SMS2. SMS2 could be a potential target for treating atherosclerosis.

De Novo Biosynthesis of Glutarate via α-Keto Acid Carbon Chain Extension and Decarboxylation Pathway in Escherichia coli.

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Wang, Jian; Wu, Yifei; Sun, Xinxiao; Yuan, Qipeng; Yan, Yajun

2017-10-20

Microbial based bioplastics are promising alternatives to petroleum based synthetic plastics due to their renewability and economic feasibility. Glutarate is one of the most potential building blocks for bioplastics. The recent biosynthetic routes for glutarate were mostly based on the l-lysine degradation pathway from Pseudomonas putida that required lysine either by feeding or lysine overproduction via genetic manipulations. Herein, we established a novel glutarate biosynthetic pathway by incorporation of a "+1" carbon chain extension pathway from α-ketoglutarate (α-KG) in combination with α-keto acid decarboxylation pathway in Escherichia coli. Introduction of homocitrate synthase (HCS), homoaconitase (HA) and homoisocitrate dehydrogenase (HICDH) from Saccharomyces cerevisiae into E. coli enabled "+1" carbon extension from α-KG to α-ketoadipate (α-KA), which was subsequently converted into glutarate by a promiscuous α-keto acid decarboxylase (KivD) and a succinate semialdehyde dehydrogenase (GabD). The recombinant E. coli coexpressing all five genes produced 0.3 g/L glutarate from glucose. To further improve the titers, α-KG was rechanneled into carbon chain extension pathway via the clustered regularly interspersed palindromic repeats system mediated interference (CRISPRi) of essential genes sucA and sucB in tricarboxylic acid (TCA) cycle. The final strain could produce 0.42 g/L glutarate, which was increased by 40% compared with the parental strain.

Mutational analysis of the RecJ exonuclease of Escherichia coli: identification of phosphoesterase motifs.

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Sutera, V A; Han, E S; Rajman, L A; Lovett, S T

1999-10-01

The recJ gene, identified in Escherichia coli, encodes a Mg(+2)-dependent 5'-to-3' exonuclease with high specificity for single-strand DNA. Genetic and biochemical experiments implicate RecJ exonuclease in homologous recombination, base excision, and methyl-directed mismatch repair. Genes encoding proteins with strong similarities to RecJ have been found in every eubacterial genome sequenced to date, with the exception of Mycoplasma and Mycobacterium tuberculosis. Multiple genes encoding proteins similar to RecJ are found in some eubacteria, including Bacillus and Helicobacter, and in the archaea. Among this divergent set of sequences, seven conserved motifs emerge. We demonstrate here that amino acids within six of these motifs are essential for both the biochemical and genetic functions of E. coli RecJ. These motifs may define interactions with Mg(2+) ions or substrate DNA. A large family of proteins more distantly related to RecJ is present in archaea, eubacteria, and eukaryotes, including a hypothetical protein in the MgPa adhesin operon of Mycoplasma, a domain of putative polyA polymerases in Synechocystis and Aquifex, PRUNE of Drosophila, and an exopolyphosphatase (PPX1) of Saccharomyces cereviseae. Because these six RecJ motifs are shared between exonucleases and exopolyphosphatases, they may constitute an ancient phosphoesterase domain now found in all kingdoms of life.

Screening of β-Glucosidase and β-Xylosidase Activities in Four Non-Saccharomyces Yeast Isolates.

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López, María Consuelo; Mateo, José Juan; Maicas, Sergi

2015-08-01

The finding of new isolates of non-Saccharomyces yeasts, showing beneficial enzymes (such as β-glucosidase and β-xylosidase), can contribute to the production of quality wines. In a selection and characterization program, we have studied 114 isolates of non-Saccharomyces yeasts. Four isolates were selected because of their both high β-glucosidase and β-xylosidase activities. The ribosomal D1/D2 regions were sequenced to identify them as Pichia membranifaciens Pm7, Hanseniaspora vineae Hv3, H. uvarum Hu8, and Wickerhamomyces anomalus Wa1. The induction process was optimized to be carried on YNB-medium supplemented with 4% xylan, inoculated with 106 cfu/mL and incubated 48 h at 28 °C without agitation. Most of the strains had a pH optimum of 5.0 to 6.0 for both the β-glucosidase and β-xylosidase activities. The effect of sugars was different for each isolate and activity. Each isolate showed a characteristic set of inhibition, enhancement or null effect for β-glucosidase and β-xylosidase. The volatile compounds liberated from wine incubated with each of the 4 yeasts were also studied, showing an overall terpene increase (1.1 to 1.3-folds) when wines were treated with non-Saccharomyces isolates. In detail, terpineol, 4-vinyl-phenol and 2-methoxy-4-vinylphenol increased after the addition of Hanseniaspora isolates. Wines treated with Hanseniaspora, Wickerhamomyces, or Pichia produced more 2-phenyl ethanol than those inoculated with other yeasts. © 2015 Institute of Food Technologists®

A recombinant CYP11B1 dependent Escherichia coli biocatalyst for selective cortisol production and optimization towards a preparative scale.

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Schiffer, Lina; Anderko, Simone; Hobler, Anna; Hannemann, Frank; Kagawa, Norio; Bernhardt, Rita

2015-02-25

Human mitochondrial CYP11B1 catalyzes a one-step regio- and stereoselective 11β-hydroxylation of 11-deoxycortisol yielding cortisol which constitutes not only the major human stress hormone but also represents a commercially relevant therapeutic drug due to its anti-inflammatory and immunosuppressive properties. Moreover, it is an important intermediate in the industrial production of synthetic pharmaceutical glucocorticoids. CYP11B1 thus offers a great potential for biotechnological application in large-scale synthesis of cortisol. Because of its nature as external monooxygenase, CYP11B1-dependent steroid hydroxylation requires reducing equivalents which are provided from NADPH via a redox chain, consisting of adrenodoxin reductase (AdR) and adrenodoxin (Adx). We established an Escherichia coli based whole-cell system for selective cortisol production from 11-deoxycortisol by recombinant co-expression of the demanded 3 proteins. For the subsequent optimization of the whole-cell activity 3 different approaches were pursued: Firstly, CYP11B1 expression was enhanced 3.3-fold to 257 nmol∗L(-1) by site-directed mutagenesis of position 23 from glycine to arginine, which was accompanied by a 2.6-fold increase in cortisol yield. Secondly, the electron transfer chain was engineered in a quantitative manner by introducing additional copies of the Adx cDNA in order to enhance Adx expression on transcriptional level. In the presence of 2 and 3 copies the initial linear conversion rate was greatly accelerated and the final product concentration was improved 1.4-fold. Thirdly, we developed a screening system for directed evolution of CYP11B1 towards higher hydroxylation activity. A culture down-scale to microtiter plates was performed and a robot-assisted, fluorescence-based conversion assay was applied for the selection of more efficient mutants from a random library. Under optimized conditions a maximum productivity of 0.84 g cortisol∗L(-1)∗d(-1) was achieved, which

Clinical Efficacy Comparison of Saccharomyces Boulardii and Lactic Acid as Probiotics in Acute Pediatric Diarrhea.

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Asmat, Shakila; Shaukat, Fouzia; Asmat, Raheela; Bakhat, Hafiz Faiq Siddique Gul; Asmat, Tauseef M

2018-03-01

To compare the efficacy of Saccharomyces boulardii and lactic acid producing probiotics in addition to usual treatment regimen to cure diarrhea among children (6 months to 5 years of age). Randomized controlled trial. Department of Pediatrics, Sheikh Zayed Hospital, Lahore, from February to July 2015. Children suffering from acute diarrhea were orally administered Saccharomyces boulardii and lactic acid producing probiotics for 5 days. The efficacy of administered probiotics was monitored. Patients were given Saccharomyces boulardii and lactic acid producing probiotics randomly to remove the bias. Two hundred patients randomly selected for trials; out of which, 100 were treated with Saccharomyces boulardii while the other 100 were supplemented with lactic acid concomitantly along with conventional diarrhea treatment. Results indicated that Saccharomyces boulardii treatment group has significantly higher efficacy rate (45%) compared to lactic acid producing probiotics (26%). This study concluded that Saccharomyces boulardii has a better efficacy compared to lactic acid and may be adopted as a probiotic of choice.

Amino acid sequences of ribosomal proteins S11 from Bacillus stearothermophilus and S19 from Halobacterium marismortui. Comparison of the ribosomal protein S11 family.

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Kimura, M; Kimura, J; Hatakeyama, T

1988-11-21

The complete amino acid sequences of ribosomal proteins S11 from the Gram-positive eubacterium Bacillus stearothermophilus and of S19 from the archaebacterium Halobacterium marismortui have been determined. A search for homologous sequences of these proteins revealed that they belong to the ribosomal protein S11 family. Homologous proteins have previously been sequenced from Escherichia coli as well as from chloroplast, yeast and mammalian ribosomes. A pairwise comparison of the amino acid sequences showed that Bacillus protein S11 shares 68% identical residues with S11 from Escherichia coli and a slightly lower homology (52%) with the homologous chloroplast protein. The halophilic protein S19 is more related to the eukaryotic (45-49%) than to the eubacterial counterparts (35%).

Escherichia coli pathotypes

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Escherichia coli strains are important commensals of the intestinal tract of humans and animals; however, pathogenic strains, including diarrhea-inducing E. coli and extraintestinal pathogenic E. coli. Intestinal E. coli pathotypes may cause a dehydrating watery diarrhea, or more severe diseases su...

Enological characterization of Spanish Saccharomyces kudriavzevii strains, one of the closest relatives to parental strains of winemaking and brewing Saccharomyces cerevisiae × S. kudriavzevii hybrids.

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Peris, D; Pérez-Través, L; Belloch, C; Querol, A

2016-02-01

Wine fermentation and innovation have focused mostly on Saccharomyces cerevisiae strains. However, recent studies have shown that other Saccharomyces species can also be involved in wine fermentation or are useful for wine bouquet, such as Saccharomyces uvarum and Saccharomyces paradoxus. Many interspecies hybrids have also been isolated from wine fermentation, such as S. cerevisiae × Saccharomyces kudriavzevii hybrids. In this study, we explored the genetic diversity and fermentation performance of Spanish S. kudriavzevii strains, which we compared to other S. kudriavzevii strains. Fermentations of red and white grape musts were performed, and the phenotypic differences between Spanish S. kudriavzevii strains under different temperature conditions were examined. An ANOVA analysis suggested striking similarity between strains for glycerol and ethanol production, although a high diversity of aromatic profiles among fermentations was found. The sources of these phenotypic differences are not well understood and require further investigation. Although the Spanish S. kudriavzevii strains showed desirable properties, particularly must fermentations, the quality of their wines was no better than those produced with a commercial S. cerevisiae. We suggest hybridization or directed evolution as methods to improve and innovate wine. Copyright © 2015 Elsevier Ltd. All rights reserved.

Antibiotic-Resistant Pathogenic Escherichia Coli Isolated from Rooftop Rainwater-Harvesting Tanks in the Eastern Cape, South Africa

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Mokaba Shirley Malema

2018-05-01

Full Text Available Although many developing countries use harvested rainwater (HRW for drinking and other household purposes, its quality is seldom monitored. Continuous assessment of the microbial quality of HRW would ensure the safety of users of such water. The current study investigated the prevalence of pathogenic Escherichia coli strains and their antimicrobial resistance patterns in HRW tanks in the Eastern Cape, South Africa. Rainwater samples were collected weekly between June and September 2016 from 11 tanks in various areas of the province. Enumeration of E. coli was performed using the Colilert®18/Quanti-Tray® 2000 method. E. coli isolates were obtained and screened for their virulence potentials using polymerase chain reaction (PCR, and subsequently tested for antibiotic resistance using the disc-diffusion method against 11 antibiotics. The pathotype most detected was the neonatal meningitis E. coli (NMEC (ibeA 28% while pathotype enteroaggregative E. coli (EAEC was not detected. The highest resistance of the E. coli isolates was observed against Cephalothin (76%. All tested pathotypes were susceptible to Gentamicin, and 52% demonstrated multiple-antibiotic resistance (MAR. The results of the current study are of public health concern since the use of untreated harvested rainwater for potable purposes may pose a risk of transmission of pathogenic and antimicrobial-resistant E. coli.

Antibiotic-Resistant Pathogenic Escherichia Coli Isolated from Rooftop Rainwater-Harvesting Tanks in the Eastern Cape, South Africa.

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Malema, Mokaba Shirley; Abia, Akebe Luther King; Tandlich, Roman; Zuma, Bonga; Mwenge Kahinda, Jean-Marc; Ubomba-Jaswa, Eunice

2018-05-01

Although many developing countries use harvested rainwater (HRW) for drinking and other household purposes, its quality is seldom monitored. Continuous assessment of the microbial quality of HRW would ensure the safety of users of such water. The current study investigated the prevalence of pathogenic Escherichia coli strains and their antimicrobial resistance patterns in HRW tanks in the Eastern Cape, South Africa. Rainwater samples were collected weekly between June and September 2016 from 11 tanks in various areas of the province. Enumeration of E. coli was performed using the Colilert ® 18/Quanti-Tray ® 2000 method. E. coli isolates were obtained and screened for their virulence potentials using polymerase chain reaction (PCR), and subsequently tested for antibiotic resistance using the disc-diffusion method against 11 antibiotics. The pathotype most detected was the neonatal meningitis E. coli (NMEC) ( ibeA 28%) while pathotype enteroaggregative E. coli (EAEC) was not detected. The highest resistance of the E. coli isolates was observed against Cephalothin (76%). All tested pathotypes were susceptible to Gentamicin, and 52% demonstrated multiple-antibiotic resistance (MAR). The results of the current study are of public health concern since the use of untreated harvested rainwater for potable purposes may pose a risk of transmission of pathogenic and antimicrobial-resistant E. coli.

Purification and Characterization of Recombinant Vaccinia L1R Protein from Escherichia coli

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2016-08-01

RECOMBINANT VACCINIA L1R PROTEIN FROM ESCHERICHIA COLI 1. INTRODUCTION 1.1 Background Vaccinia virus (VACV) is the active component of the...the preparation of the recombinant VACV L1R protein fragment by denaturing , refolding, and purifying material expressed into inclusion bodies in...PURIFICATION AND CHARACTERIZATION OF RECOMBINANT VACCINIA L1R PROTEIN FROM ESCHERICHIA COLI ECBC-TR-1370

Humanized HLA-DR4.RagKO.IL2RγcKO.NOD (DRAG) mice sustain the complex vertebrate life cycle of Plasmodium falciparum malaria.

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Wijayalath, Wathsala; Majji, Sai; Villasante, Eileen F; Brumeanu, Teodor D; Richie, Thomas L; Casares, Sofia

2014-09-30

Malaria is a deadly infectious disease affecting millions of people in tropical and sub-tropical countries. Among the five species of Plasmodium parasites that infect humans, Plasmodium falciparum accounts for the highest morbidity and mortality associated with malaria. Since humans are the only natural hosts for P. falciparum, the lack of convenient animal models has hindered the understanding of disease pathogenesis and prompted the need of testing anti-malarial drugs and vaccines directly in human trials. Humanized mice hosting human cells represent new pre-clinical models for infectious diseases that affect only humans. In this study, the ability of human-immune-system humanized HLA-DR4.RagKO.IL2RγcKO.NOD (DRAG) mice to sustain infection with P. falciparum was explored. Four week-old DRAG mice were infused with HLA-matched human haematopoietic stem cells (HSC) and examined for reconstitution of human liver cells and erythrocytes. Upon challenge with infectious P. falciparum sporozoites (NF54 strain) humanized DRAG mice were examined for liver stage infection, blood stage infection, and transmission to Anopheles stephensi mosquitoes. Humanized DRAG mice reconstituted human hepatocytes, Kupffer cells, liver endothelial cells, and erythrocytes. Upon intravenous challenge with P. falciparum sporozoites, DRAG mice sustained liver to blood stage infection (average 3-5 parasites/microlitre blood) and allowed transmission to An. stephensi mosquitoes. Infected DRAG mice elicited antibody and cellular responses to the blood stage parasites and self-cured the infection by day 45 post-challenge. DRAG mice represent the first human-immune-system humanized mouse model that sustains the complex vertebrate life cycle of P. falciparum without the need of exogenous injection of human hepatocytes/erythrocytes or P. falciparum parasite adaptation. The ability of DRAG mice to elicit specific human immune responses to P. falciparum parasites may help deciphering immune correlates

Identification of mitochondrial carriers in Saccharomyces cerevisiae by transport assay of reconstituted recombinant proteins.

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Palmieri, Ferdinando; Agrimi, Gennaro; Blanco, Emanuela; Castegna, Alessandra; Di Noia, Maria A; Iacobazzi, Vito; Lasorsa, Francesco M; Marobbio, Carlo M T; Palmieri, Luigi; Scarcia, Pasquale; Todisco, Simona; Vozza, Angelo; Walker, John

2006-01-01

The inner membranes of mitochondria contain a family of carrier proteins that are responsible for the transport in and out of the mitochondrial matrix of substrates, products, co-factors and biosynthetic precursors that are essential for the function and activities of the organelle. This family of proteins is characterized by containing three tandem homologous sequence repeats of approximately 100 amino acids, each folded into two transmembrane alpha-helices linked by an extensive polar loop. Each repeat contains a characteristic conserved sequence. These features have been used to determine the extent of the family in genome sequences. The genome of Saccharomyces cerevisiae contains 34 members of the family. The identity of five of them was known before the determination of the genome sequence, but the functions of the remaining family members were not. This review describes how the functions of 15 of these previously unknown transport proteins have been determined by a strategy that consists of expressing the genes in Escherichia coli or Saccharomyces cerevisiae, reconstituting the gene products into liposomes and establishing their functions by transport assay. Genetic and biochemical evidence as well as phylogenetic considerations have guided the choice of substrates that were tested in the transport assays. The physiological roles of these carriers have been verified by genetic experiments. Various pieces of evidence point to the functions of six additional members of the family, but these proposals await confirmation by transport assay. The sequences of many of the newly identified yeast carriers have been used to characterize orthologs in other species, and in man five diseases are presently known to be caused by defects in specific mitochondrial carrier genes. The roles of eight yeast mitochondrial carriers remain to be established.

Application of analytic hierarchy process in landscape management: Case study area Košutnjak park-forest

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Lakićević Milena

2011-01-01

Full Text Available Proper management is one of key elements of the natural landscape protection strategy. Park-forests with protected and conserved natural elements represent attractive eco-tourism urban zones. Košutnjak is the most visited park-forest in Serbian capitol Belgrade, unfortunately with increasing number of degraded and devastated areas as a consequence of negative human impacts in the past. In order to conserve natural values in this popular forested city area, and to improve its tourism potential, we found that in achieving that goal, it is meaningful to asses possible management practices and identify the most desired one by applying the analytic hierarchy process (AHP, scientifically sound multi-criteria decision making tool. Based on the obtained results, a recommended strategy is to renovate natural vegetation and to promote recreational and tourism offer in Košutnjak with respect of the sustainability principle.

Accumulation of gold using Baker's yeast, Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Roy, Kamalika; Lahiri, Susanta; Sinha, P.

2006-01-01

Authors have reported preconcentration of 152 Eu, a long-lived fission product, by yeast cells, Saccharomyces cerevisiae. Gold being a precious metal is used in electroplating, hydrogenation catalyst, etc. Heterogeneous composition of samples and low concentration offers renewed interest in its selective extraction of gold using various extractants. Gold can be recovered from different solutions using various chemical reagents like amines, organophosphorus compounds, and extractants containing sulphur as donor atom, etc. In the present work, two different strains of baker's yeast, Saccharomyces cerevisiae have been used to study the preconcentration of gold at various experimental conditions

Use of non-saccharomyces Torulaspora delbrueckii yeast strains in winemaking and brewing

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Tataridis Panagiotis

2013-01-01

Full Text Available Selected Saccharomyces yeast strains have been used for more than 150 years in brewing and for several decades in winemaking. They are necessary in brewing because of the boiling of the wort, which results in the death of all yeast cells, with the exception of some Belgian style beers (ex. Lambic, where the wort is left to be colonized by indigenous yeast and bacteria from the environment and ferment naturally. In winemaking their use is also pertinent because they provide regular and timely fermentations, inhibit the growth of indigenous spoilage microorganisms and contribute to the desired sensory characters. Even though the use of selected Saccharomyces strains provides better quality assurance in winemaking in comparison to the unknown microbial consortia in the must, it has been debated for a long time now whether the use of selected industrial Saccharomyces strains results in wines with less sensory complexity and “terroir” character. In previous decades, non-Saccharomyces yeasts were mainly considered as spoilage/problematic yeast, since they exhibited low fermentation ability and other negative traits. In the last decades experiments have shown that there are some non-Saccharomyces strains (Candida, Pichia, Kluyveromyces, Torulaspora, etc which, even though they are not able to complete the fermentation they can still be used in sequential inoculation-fermentation with Saccharomyces to increase sensory complexity of the wines. Through fermentation in a laboratory scale, we have observed that the overall effects of selected Torulaspora delbrueckii yeast strains, is highly positive, leading to products with pronounced sensory complexity and floral/fruity aroma in winemaking and brewing.

Non-Saccharomyces Yeasts Nitrogen Source Preferences: Impact on Sequential Fermentation and Wine Volatile Compounds Profile

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Antoine Gobert

2017-11-01

Full Text Available Nitrogen sources in the must are important for yeast metabolism, growth, and performance, and wine volatile compounds profile. Yeast assimilable nitrogen (YAN deficiencies in grape must are one of the main causes of stuck and sluggish fermentation. The nitrogen requirement of Saccharomyces cerevisiae metabolism has been described in detail. However, the YAN preferences of non-Saccharomyces yeasts remain unknown despite their increasingly widespread use in winemaking. Furthermore, the impact of nitrogen consumption by non-Saccharomyces yeasts on YAN availability, alcoholic performance and volatile compounds production by S. cerevisiae in sequential fermentation has been little studied. With a view to improving the use of non-Saccharomyces yeasts in winemaking, we studied the use of amino acids and ammonium by three strains of non-Saccharomyces yeasts (Starmerella bacillaris, Metschnikowia pulcherrima, and Pichia membranifaciens in grape juice. We first determined which nitrogen sources were preferentially used by these yeasts in pure cultures at 28 and 20°C (because few data are available. We then carried out sequential fermentations at 20°C with S. cerevisiae, to assess the impact of the non-Saccharomyces yeasts on the availability of assimilable nitrogen for S. cerevisiae. Finally, 22 volatile compounds were quantified in sequential fermentation and their levels compared with those in pure cultures of S. cerevisiae. We report here, for the first time, that non-Saccharomyces yeasts have specific amino-acid consumption profiles. Histidine, methionine, threonine, and tyrosine were not consumed by S. bacillaris, aspartic acid was assimilated very slowly by M. pulcherrima, and glutamine was not assimilated by P. membranifaciens. By contrast, cysteine appeared to be a preferred nitrogen source for all non-Saccharomyces yeasts. In sequential fermentation, these specific profiles of amino-acid consumption by non-Saccharomyces yeasts may account for

Non-Saccharomyces Yeasts Nitrogen Source Preferences: Impact on Sequential Fermentation and Wine Volatile Compounds Profile

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Gobert, Antoine; Tourdot-Maréchal, Raphaëlle; Morge, Christophe; Sparrow, Céline; Liu, Youzhong; Quintanilla-Casas, Beatriz; Vichi, Stefania; Alexandre, Hervé

2017-01-01

Nitrogen sources in the must are important for yeast metabolism, growth, and performance, and wine volatile compounds profile. Yeast assimilable nitrogen (YAN) deficiencies in grape must are one of the main causes of stuck and sluggish fermentation. The nitrogen requirement of Saccharomyces cerevisiae metabolism has been described in detail. However, the YAN preferences of non-Saccharomyces yeasts remain unknown despite their increasingly widespread use in winemaking. Furthermore, the impact of nitrogen consumption by non-Saccharomyces yeasts on YAN availability, alcoholic performance and volatile compounds production by S. cerevisiae in sequential fermentation has been little studied. With a view to improving the use of non-Saccharomyces yeasts in winemaking, we studied the use of amino acids and ammonium by three strains of non-Saccharomyces yeasts (Starmerella bacillaris, Metschnikowia pulcherrima, and Pichia membranifaciens) in grape juice. We first determined which nitrogen sources were preferentially used by these yeasts in pure cultures at 28 and 20°C (because few data are available). We then carried out sequential fermentations at 20°C with S. cerevisiae, to assess the impact of the non-Saccharomyces yeasts on the availability of assimilable nitrogen for S. cerevisiae. Finally, 22 volatile compounds were quantified in sequential fermentation and their levels compared with those in pure cultures of S. cerevisiae. We report here, for the first time, that non-Saccharomyces yeasts have specific amino-acid consumption profiles. Histidine, methionine, threonine, and tyrosine were not consumed by S. bacillaris, aspartic acid was assimilated very slowly by M. pulcherrima, and glutamine was not assimilated by P. membranifaciens. By contrast, cysteine appeared to be a preferred nitrogen source for all non-Saccharomyces yeasts. In sequential fermentation, these specific profiles of amino-acid consumption by non-Saccharomyces yeasts may account for some of the

PF-KO system for single bunch mode operation of a storage ring

International Nuclear Information System (INIS)

Ohgaki, H.; Sugiyama, S.; Mikado, T.; Chiwaki, M.; Yamada, K.; Suzuki, R.; Sei, N.; Noguchi, T.; Yamazaki, T.

1994-01-01

A new RF-KO (RF knockout) system for the single bunch mode operation of a storage ring has been developed. The knockout signal is modulated by the sum signal of the RF acceleration frequency of the storage ring and a bunch selection signal. We do not need any special device or a timing unit with this method. We obtain a high purity of bunch structure in a short knock out time. The single bunch impurity of 0.2% has been achieved. (author)

Studies of N ~ 40 Ni isotopes via neutron-knockout (nKO) and deep-inelastic (DI) reactions

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Chiara, C. J.; Recchia, F.; Gade, A.; Janssens, R. V. F.; Walters, W. B.

2013-10-01

V. BADER, T. BAUGHER, D. BAZIN, J.S. BERRYMAN, B.A. BROWN, C. LANGER, N. LARSON, S.N. LIDDICK, E. LUNDERBERG, S. NOJI, C. PROKOP, S.R. STROBERG, S. SUCHYTA, D. WEISSHAAR, S. WILLIAMS, NSCL/MSU, M. ALBERS, M. ALCORTA, P.F. BERTONE, M.P. CARPENTER, J. CHEN, C.R. HOFFMAN, F.G. KONDEV, T. LAURITSEN, A.M. ROGERS, D. SEWERYNIAK, S. ZHU, ANL, C.M. CAMPBELL, LBNL, H.M. DAVID, D.T. DOHERTY, U. of Edinburgh/ANL, A. KORICHI, CSNSM-IN2P3/ANL, C.J. LISTER, U. of Mass.-Lowell, K. WIMMER, Central Mich. U. -- Excited states in 68Ni were populated in 2nKO reactions at NSCL. Prompt γ rays were detected with the GRETINA array located in front of the S800 separator. A hodoscope at the S800 focal plane captured the 68Ni ions, where isomeric decays could be correlated with prompt γ rays. Decay of the first excited state, a 0+ isomer, was observed, confirming that its energy substantially differs from the literature value. Comparing the decay patterns of excited states with shell-model calculations provides insight into their underlying structure. Data from 70Zn + 208Pb DI reactions studied with Gammasphere provide results consistent with the 2nKO. Single-particle strengths are also under investigation in the odd- A Ni isotopes via 1nKO reactions. Supported in part by the DoE (DE-FG02-94ER40834, DE-AC02-06CH11357), NSF (PHY-1102511), and NNSA (DE-NA0000979).

Mitochondrial genome evolution in the Saccharomyces sensu stricto complex.

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Ruan, Jiangxing; Cheng, Jian; Zhang, Tongcun; Jiang, Huifeng

2017-01-01

Exploring the evolutionary patterns of mitochondrial genomes is important for our understanding of the Saccharomyces sensu stricto (SSS) group, which is a model system for genomic evolution and ecological analysis. In this study, we first obtained the complete mitochondrial sequences of two important species, Saccharomyces mikatae and Saccharomyces kudriavzevii. We then compared the mitochondrial genomes in the SSS group with those of close relatives, and found that the non-coding regions evolved rapidly, including dramatic expansion of intergenic regions, fast evolution of introns and almost 20-fold higher rearrangement rates than those of the nuclear genomes. However, the coding regions, and especially the protein-coding genes, are more conserved than those in the nuclear genomes of the SSS group. The different evolutionary patterns of coding and non-coding regions in the mitochondrial and nuclear genomes may be related to the origin of the aerobic fermentation lifestyle in this group. Our analysis thus provides novel insights into the evolution of mitochondrial genomes.

PRODUKSI ETANOL DARI TETES TEBU OLEH Saccharomyces cerevisiae PEMBENTUK FLOK (NRRL – Y 265 (Ethanol Production from Cane Molasses by Flocculant Saccharomyces cerevisiae (NRRL – Y 265

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Agustin Krisna Wardani

2013-08-01

Full Text Available The potential use of sugar cane molasses by flocculant Saccharomyces cerevisiae in ethanol production was investigated. In order to minimize the negative effect of calcium on yeast growth, pretreated sugar cane molasses with dilute acid was performed. The influence of process parameters such as sugar concentration and inoculum concentration were evaluated for enhancing bioethanol production. Result showed that maximum ethanol concentration of 8,792% (b/v was obtained at the best condition of inoculum concentration 10% (v/v and sugar concentration 15% (b/v. Based on the experimental data, maximum yield of ethanol production of 65% was obtained. This result demonstrated the potential of molasses as promising biomass resources for ethanol production. Keywords: Ethanol, preteated cane molasses, flocculant Saccharomyces cerevisiae, fermentation   ABSTRAK Efisiensi produksi bioetanol diperoleh melalui ketepatan pemilihan jenis mikroorganisme, bahan baku, dan kontrol proses fermentasi. Alternatif proses untuk meminimalisasi biaya produksi etanol adalah dengan mengeliminasi tahap pemisahan sentrifugasi sel dari produk karena memerlukan biaya instalasi dan biaya perawatan yang tinggi. Proses sentrifugasi merupakan tahapan penting untuk memisahkan sel mikroba dari medium fermentasi pada produksi bioetanol. Untuk meminimalisir biaya produksi akibat proses tersebut digunakan inokulum Saccharomyces cerevisiae pembentuk flok dan tetes tebu sebagai sumber gula. Penelitian ini bertujuan untuk mendapatkan konsentrasi penambahan inokulum Saccharomyces cerevisiae pembentuk flok dan konsentrasi sumber gula dalam tetes tebu yang tepat dalam produksi etanol yang maksimum. Saccharomyces cerevisiae sebanyak 5%, 10%, dan 15% (v/v diinokulasikan pada medium tetes tebu hasil pretreatment dengan kandungan gula 15%, 20%, dan 25% (b/v pada pH 5. Fermentasi dilakukan pada suhu 30°C dan agitasi 100 rpm selama 72 jam. Etanol tertinggi didapat pada kondisi konsentrasi inokulum

Genetic and phenotypic characterization of Saccharomyces spp. strains isolated in distillery plants.

Science.gov (United States)

Úbeda, Juan F; Chacón-Ocaña, Maria; Díaz-Hellín, Patricia; Ramírez-Pérez, Hector; Briones, Ana

2016-06-01

In this study, the biodiversity and some interesting phenotypic properties of Saccharomyces wild yeasts isolated in distilleries, at least 100 years old, located in La Mancha (Spain), were determined. Strains were genetically characterized by RFLP-mtDNA, which confirmed a great genetic biodiversity with 73% of strains with different mtDNA profiles, highlighting the large variability found in sweet and fermented piquette substrata. The predominant species identified was S. cerevisiae, followed by S. paradoxus and S. bayanus Due to the residual sugar-alcohol extraction process using warm water, a great number of thermophilic Saccharomyces strains with a great cell vitality were found to have potential use as starters in distillery plants. Interesting technological properties such as cell vitality and growth rate at different temperatures were studied. The thermal washing process for the extraction of alcohol and reducing sugars of some raw materials contributes to the presence of Saccharomyces strains with technologically interesting properties, especially in terms of vitality and resistance to high temperatures. Due to the fact that fermentation is spontaneous, the yeast biota of these environments, Saccharomyces and non-Saccharomyces, is very varied so these ecological niches are microbial reserves of undoubted biotechnological interest. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Functional consequences of brain glycogen deficiency on the sleep-wake cycle regulation in PTG-KO mice

KAUST Repository

Burlet-Godinot, S.; Allaman, I.; Grenningloh, G.; Roach, P.J.; Depaoli-Roach, A.A.; Magistretti, Pierre J.; Petit, J.-M.

2017-01-01

in the brain in mice while glycogen levels were paradoxically maintained and not affected. In order to gain further insight on the role of PTG in this process, we studied the sleep/wake cycle parameters in PTG knockout (PTG-KO) mice under baseline conditions

REKAYASA BIOPROSES PRODUKSI BIOETANOL DARI UBI KAYU DENGAN TEKNIK KO-KULTUR RAGI TAPE DAN SACCHAROMYCES CEREVISIAE

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IWayan Arnata

2016-11-01

Full Text Available The purpose of this study is to get an alternative bioprocess technology of bioethanolproduction from cassava using the technique of co-cultured “tape” yeast and Saccharomycescerevisiae. The study was designed with a completely randomized design experiment using 7level of co-culture techniques. All treatment was repeated 2 times. The parameters measuredwere the final pH, percentage of substrate consumption, ethanol concentration and fermentationefficiency. Using coculture technique in the process of fermentation to producebioethanol gives better results than the use of a single culture or pure culture of “tape”yeast and S. cerevisiae. Coculture technique that gives the best results is the treatmentof fermentation process by giving the “tape” yeast to the first day and followed bygiving culture S. cerevisiae for the next two days. This treatment has a final pH 4:05,the percentage substrate or glucose consumption of 82.92%, the concentration ethanolof 11.0% (w/v and the fermentation efficiency of 52.94%.

Abnormal nuclear envelope in the cerebellar Purkinje cells and impaired motor learning in DYT11 myoclonus-dystonia mouse models

OpenAIRE

Yokoi, Fumiaki; Dang, Mai T.; Yang, Guang; Li, JinDong; Doroodchi, Atbin; Zhou, Tong; Li, Yuqing

2011-01-01

Myoclonus-dystonia (M-D) is a movement disorder characterized by myoclonic jerks with dystonia. DYT11 M-D is caused by mutations in SGCE which codes for ε-sarcoglycan. SGCE is maternally imprinted and paternally expressed. Abnormal nuclear envelope has been reported in mouse models of DYT1 generalized torsion dystonia. However, it is not known whether similar alterations occur in DYT11 M-D. We developed a mouse model of DYT11 M-D using paternally-inherited Sgce heterozygous knockout (Sgce KO)...

Influence of a Saccharomyces cerevisia fermentation product on the pathophysiological response to a combined intranasal bovine herpesvirus-1 and intratracheal Mannheimia haemolytica challenge in Holstein steers

Science.gov (United States)

The objective of this study was to determine the effects of supplementing a Saccharomyces cerevisiae fermentation product prototype (Prototype) on the pathophysiological response during a combined viral-bacterial respiratory challenge. Holstein steer calves (126.5±6.11kg; N=16) were completely rando...

Multiple antimicrobial resistance of Escherichia coli and Salmonella ...

African Journals Online (AJOL)

Presumptive isolates were subjected to antimicrobial susceptibility testing using 13 panels of antibiotics for both E. coli and Salmonella spp. Results showed that the overall isolation rate of Salmonella spp. was 12 (11.4%), broiler chickens had higher isolation rate 9 (12.0%) of Salmonella than local chickens. However, the ...

Hybridization and adaptive evolution of diverse Saccharomyces species for cellulosic biofuel production.

Science.gov (United States)

Peris, David; Moriarty, Ryan V; Alexander, William G; Baker, EmilyClare; Sylvester, Kayla; Sardi, Maria; Langdon, Quinn K; Libkind, Diego; Wang, Qi-Ming; Bai, Feng-Yan; Leducq, Jean-Baptiste; Charron, Guillaume; Landry, Christian R; Sampaio, José Paulo; Gonçalves, Paula; Hyma, Katie E; Fay, Justin C; Sato, Trey K; Hittinger, Chris Todd

2017-01-01

Lignocellulosic biomass is a common resource across the globe, and its fermentation offers a promising option for generating renewable liquid transportation fuels. The deconstruction of lignocellulosic biomass releases sugars that can be fermented by microbes, but these processes also produce fermentation inhibitors, such as aromatic acids and aldehydes. Several research projects have investigated lignocellulosic biomass fermentation by the baker's yeast Saccharomyces cerevisiae . Most projects have taken synthetic biological approaches or have explored naturally occurring diversity in S. cerevisiae to enhance stress tolerance, xylose consumption, or ethanol production. Despite these efforts, improved strains with new properties are needed. In other industrial processes, such as wine and beer fermentation, interspecies hybrids have combined important traits from multiple species, suggesting that interspecies hybridization may also offer potential for biofuel research. To investigate the efficacy of this approach for traits relevant to lignocellulosic biofuel production, we generated synthetic hybrids by crossing engineered xylose-fermenting strains of S. cerevisiae with wild strains from various Saccharomyces species. These interspecies hybrids retained important parental traits, such as xylose consumption and stress tolerance, while displaying intermediate kinetic parameters and, in some cases, heterosis (hybrid vigor). Next, we exposed them to adaptive evolution in ammonia fiber expansion-pretreated corn stover hydrolysate and recovered strains with improved fermentative traits. Genome sequencing showed that the genomes of these evolved synthetic hybrids underwent rearrangements, duplications, and deletions. To determine whether the genus Saccharomyces contains additional untapped potential, we screened a genetically diverse collection of more than 500 wild, non-engineered Saccharomyces isolates and uncovered a wide range of capabilities for traits relevant to

ISOTERMAS DE ADSORÇÃO DE CÁDMIO POR Saccharomyces cerevisiae ISOTHERMS OF CADMIUM ADSORPTION BY Saccharomyces cerevisae

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Silvana ALBERTINI

2001-08-01

Full Text Available Com o objetivo de determinar as isotermas de adsorção de cádmio por Saccharomyces cerevisiae, foram utilizados os sais cloreto e nitrato de cádmio nas concentrações de 5, 10, 20, 40, 60, 80 e 100mg L-1. A biomassa foi produzida a partir de uma cultura "starter"de Saccharomyces cerevisiae IZ 1904. Após o contato de 16h entre o microrganismo e as soluções em estudo, a biomassa foi separada por centrifugação e o teor de cádmio residual foi determinado no sobrenadante por espectrofotometria de absorção atômica. Para os dois sais empregados foi observado um acúmulo crescente de cádmio nas concentrações de 5, 10, 20 e 40mg L-1. Nas concentrações de 60, 80 e 100mg L-1 foi observado que a levedura acumulou teores menores do metal, evidenciando danos na parede celular, nem sempre acompanhados de iguais danos da membrana citoplasmática, tais alterações da parede visualizadas por microscopia eletrônica de varredura.With the objective of determining the isotherms of cadmium the adsorption by Saccharomyces cerevisiae, the chloride and nitrate salts were used in the concentrations of 5, 10, 20, 40, 60, 80, and 100mg L-1. The biomass was produced from a starter culture of Saccharomyces cerevisiae IZ 1904. After a 16h contact between the microrganism and solutions of study the biomass was separated by a centrifuge and the cadmium residue content was determined at the supernatant by atomic adsorption spectrophotometry. For the two salts used a growing accumulation of cadmium was observed at concentrations of 5, 10, 20, and 40mg L-1. In the concentrations of 60, 80 and 100mg L-1 a decreasing of the accumulation of the metal was observed, evidencing damages of the cellular wall, which they're not accompanied always by damages of the citoplasmatic membrane, visualized by scanning electron microscopy.

Antioxidant properties and global metabolite screening of the probiotic yeast Saccharomyces cerevisiae var. boulardii.

Science.gov (United States)

Datta, Suprama; Timson, David J; Annapure, Uday S

2017-07-01

Saccharomyces cerevisiae var. boulardii is the only yeast species with probiotic properties. It is considered to have therapeutic significance in gastrointestinal disorders. In the present study, a comparative physiological study between this yeast and Saccharomyces cerevisiae (BY4742) was performed by evaluating two prominent traits of probiotic species, responses to different stress conditions and antioxidant capacity. A global metabolite profile was also developed aiming to identify which therapeutically important secondary metabolites are produced. Saccharomyces cerevisiae var. boulardii showed no significant difference in growth patterns but greater stress tolerance compared to S. cerevisiae. It also demonstrated a six- to 10-fold greater antioxidant potential (judged by the 1,1-diphenyl-2-picrylhydrazyl assay), with a 70-fold higher total phenolic content and a 20-fold higher total flavonoid content in the extracellular fraction. These features were clearly differentiated by principal component analysis and further indicated by metabolite profiling. The extracellular fraction of the S. cerevisiae var. boulardii cultures was found to be rich in polyphenolic metabolites: vanillic acid, cinnamic acid, phenyl ethyl alcohol (rose oil), erythromycin, amphetamine and vitamin B 6 , which results in the antioxidant capacity of this strain. The present study presents a new perspective for differentiating the two genetically related strains of yeast, S. cerevisiae and S. cerevisiae var. boulardii by assessing their metabolome fingerprints. In addition to the correlation of the phenotypic properties with the secretory metabolites of these two yeasts, the present study also emphasizes the potential to exploit S. cerevisiae var. boulardii in the industrial production of these metabolites. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Saccharomyces jurei sp. nov., isolation and genetic identification of a novel yeast species from Quercus robur.

Science.gov (United States)

Naseeb, Samina; James, Stephen A; Alsammar, Haya; Michaels, Christopher J; Gini, Beatrice; Nueno-Palop, Carmen; Bond, Christopher J; McGhie, Henry; Roberts, Ian N; Delneri, Daniela

2017-06-01

Two strains, D5088T and D5095, representing a novel yeast species belonging to the genus Saccharomyces were isolated from oak tree bark and surrounding soil located at an altitude of 1000 m above sea level in Saint Auban, France. Sequence analyses of the internal transcribed spacer (ITS) region and 26S rRNA D1/D2 domains indicated that the two strains were most closely related to Saccharomyces mikatae and Saccharomyces paradoxus. Genetic hybridization analyses showed that both strains are reproductively isolated from all other Saccharomyces species and, therefore, represent a distinct biological species. The species name Saccharomyces jurei sp. nov. is proposed to accommodate these two strains, with D5088T (=CBS 14759T=NCYC 3947T) designated as the type strain.

The Role of Enteropathogenic E.coli in the Development and Progression of Chronic Non-Ulcerative Colitis

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L.I. Sydorchuk

2016-04-01

Full Text Available Objective. The analysis of the literature has shown that in patients with chronic non-ulcerative colitis, there were isolated E.coli that cause dysentery-like, cholera-like diseases and escherichioses. Thus, the aim of our study was to establish the level of persistence of enteropathogenic Escherichia in the colon of patients with chronic non-ulcerative colitis and their importance in the ecological system «macroorganism — microbiota». Material and methods. During 2000–2015, there has been conducted a bacteriological examination of the colon content in patients with chronic non-ulcerative colitis aged 27–41 years (average age 37.74 ± 3.62 years. The resulting cultures were examined in the indicative agglutination tests with OKA polyvalent serum. Positive cultures were tested with polyvalent serum of narrow spectrum ОКВ, ОКС, ОКD and ОКЕ. Results. In patients, in colon cavity of which E.coli O18ac:K77; O26:K60; O55:K59; O128ab:K67 were detected, the disease occurs by nosology of colienteritis. In those patients, in whom O25:K11; O144:K; O124:K72 serotypes have been identified, disease occurred as bacterial dysentery, and O25:K11 and O128:K67 — in chole­ra-like form. Discussion. In 94.50 % of patients, common E.coli were detected, but in 55 (35.03 % of them enteropathogenic Escherichia persist, in 41 (26.11 % — E.coli Hly+, in 37 (23.57 % — E.coli Lac–, in 18 (11.46 % — enterotoxigenic E.coli, in 14 (8.42 % — enteroinvasive, and in 11 (7.01 % — enterohemorragic Escherichia. Conclusions. It was determined that the main serotypes of opportunistic Escherichia, which colonize and persist in the colon cavity of patients with chronic non-ulcerative colitis, were O114:K90; O25:K11; O124:K72; O128:K67; O18ac:K77, the persistence of which affects clinical manifestation from colitis to dysentery-like or cholera-like disease.

Functional co-operation between the nuclei of Saccharomyces cerevisiae and mitochondria from other yeast species

DEFF Research Database (Denmark)

Spirek, M.; Horvath, A.; Piskur, Jure

2000-01-01

We elaborated a simple method that allows the transfer of mitochondria from collection yeasts to Saccharomyces cerevisiae. Protoplasts prepared from different yeasts were fused to the protoplasts of the ade2-1, ura3-52, kar1-1, rho (0) strain of S. cerevisiae and were selected for respiring cybrids....... italicus, S, oviformis, S. capensis and S. chevalieri) exhibited complete compatibility with S. cerevisiae nuclei. The closely related S. douglasii mitochondrial genome could also partially restore respiration-deficiency in rho (0) S. cerevisiae, whereas mitochondrial genomes from phylogenetically less...

Study on biosorption of uranium by alginate immobilized saccharomyces cerevisiae

International Nuclear Information System (INIS)

Wang Baoe; Xu Weichang; Xie Shuibo; Guo Yangbin

2005-01-01

Saccharomyces cerevisiae has great capability of biosorption of uranium. The maxium uptake is 172.4 mg/g according to this study. To adapt to the application of the biomass in the field, the biosorption of uranium by cross-linked and alginate calcium immobilized Saccharomyces cerevisiae is studied. Results indicate the maxium uptake is 185.2 mg/g by formaldehyde cross-linked biomass, and it is 769.2 mg/g by alginate calcium immobilized biomass. (authors)

E coli enteritis

Science.gov (United States)

... coli; Food poisoning - E. coli; E. coli diarrhea; Hamburger disease ... coleslaw or potato salad) that have been out of the refrigerator too ... reheated Fish or oysters Raw fruits or vegetables that have ...

The adsorption of Sr(II) and Cs(I) ions by irradiated Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Yiming Tan; Jundong Feng; Liang Qiu; Zhentian Zhao; Xiaohong Zhang; Haiqian Zhang

2017-01-01

Adsorption behavior and mechanism of Sr(II) and Cs(I) in single and binary solutions using irradiated Saccharomyces cerevisiae was investigated. The effects of several environmental factors on Sr(II) and Cs(I) adsorption to irradiated Saccharomyces cerevisiae was determined. The equilibrium experimental data were simulated by different kinetic models and isotherm models. The combined effect of Sr(II) and Cs(I) on Saccharomyces cerevisiae is generally antagonistic. SEM and EDS analyses indicate that crystals formed on the cell surface are precipitate of Sr(II) and Cs(I), respectively. (author)

Enteroaggregative Escherichia coli is the predominant diarrheagenic E. coli pathotype among irrigation water and food sources in South Africa.

Science.gov (United States)

Aijuka, Matthew; Santiago, Araceli E; Girón, Jorge A; Nataro, James P; Buys, Elna M

2018-08-02

Diarrheagenic E. coli (DEC) has been implicated in foodborne outbreaks worldwide and have been associated with childhood stunting in the absence of diarrhoea. Infection is extraordinarily common, but the routes of transmission have not been determined. Therefore, determining the most prevalent pathotypes in food and environmental sources may help provide better guidance to various stakeholders in ensuring food safety and public health and advancing understanding of the epidemiology of enteric disease. We characterized 205 E. coli strains previously isolated from producer distributor bulk milk (PDBM)(118), irrigation water (48), irrigated lettuce (29) and street vendor coleslaw (10) in South Africa. Enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroaggregative E. coli (EAEC) and diffusely adherent E. coli (DAEC) were sought. We used PCR and partial gene sequencing for all 205 strains while 46 out of 205 that showed poor resolution were subsequently characterized using cell adherence (HeLa cells). PCR and partial gene sequencing of aatA and/or aaiC genes confirmed EAEC (2%, 5 out of 205) as the only pathotype. Phylogenetic analysis of sequenced EAEC strains with E. coli strains in GenBank showing ≥80% nucleotide sequence similarity based on possession of aaiC and aatA generated distinct clusters of strains separated predominantly based on their source of isolation (food source or human stool) suggesting a potential role of virulence genes in source tracking. EAEC 24%, 11 out of 46 strains (PDBM = 15%, irrigation water = 7%, irrigated lettuce = 2%) was similarly the predominant pathotype followed by strains showing invasiveness to HeLa cells, 4%, 2 out of 46 (PDBM = 2%, irrigated lettuce = 2%), among stains characterized using cell adherence. Therefore, EAEC may be the leading cause of DEC associated food and water-borne enteric infection in South Africa. Additionally, solely using molecular based methods targeting virulence

Production of a heterologous proteinase A by Saccharomyces kluyveri

DEFF Research Database (Denmark)

Møller, Kasper; Tidemand, L.D.; Winther, J.R.

2001-01-01

In order to evaluate the potential of Saccharomyces kluyveri for heterologous protein production, S. kluyveri Y159 was transformed with a S. cerevisiae-based multi-copy plasmid containing the S. cerevisiae PEP4 gene, which encodes proteinase A, under the control of its native promoter. As a refer......In order to evaluate the potential of Saccharomyces kluyveri for heterologous protein production, S. kluyveri Y159 was transformed with a S. cerevisiae-based multi-copy plasmid containing the S. cerevisiae PEP4 gene, which encodes proteinase A, under the control of its native promoter...

Isolation of the thymidylate synthetase gene (TMP1) by complementation in Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Taylor, G.R.; Barclay, B.J.; Storms, R.K.; Friesen, J.D.; Haynes, R.H.

1982-01-01

The structural gene (TMP1) for yeast thymidylate synthetase (thymidylate synthase; EC 2.1.1.45) was isolated from a chimeric plasmid bank by genetic complementation in Saccharomyces cerevisiae. Retransformation of the dTMP auxotroph GY712 and a temperature-sensitive mutant (cdc21) with purified plasmid (pTL1) yielded Tmp/sup +/ transformants at high frequency. In addition, the plasmid was tested for the ability to complement a bacterial thyA mutant that lacks functional thymidylate synthetase. Although it was not possible to select Thy/sup +/ transformants directly, it was found that all pTL1 transformants were phenotypically Thy/sup +/ after several generations of growth in nonselective conditions. Thus, yeast thymidylate synthetase is biologically active in Escherichia coli. Thymidylate synthetase was assayed in yeast cell lysates by high-pressure liquid chromatography to monitor the conversion of [6-/sup 3/H]dUMP to [6-/sup 3/H]dTMP. In protein extracts from the thymidylate auxotroph (tmpl-6) enzymatic conversion of dUMP to dTMP was barely detectable. Lysates of pTL1 transformants of this strain, however, had thymidylate synthetase activity that was comparable to that of the wild-type strain

Ontology-based literature mining of E. coli vaccine-associated gene interaction networks.

Science.gov (United States)

Hur, Junguk; Özgür, Arzucan; He, Yongqun

2017-03-14

Pathogenic Escherichia coli infections cause various diseases in humans and many animal species. However, with extensive E. coli vaccine research, we are still unable to fully protect ourselves against E. coli infections. To more rational development of effective and safe E. coli vaccine, it is important to better understand E. coli vaccine-associated gene interaction networks. In this study, we first extended the Vaccine Ontology (VO) to semantically represent various E. coli vaccines and genes used in the vaccine development. We also normalized E. coli gene names compiled from the annotations of various E. coli strains using a pan-genome-based annotation strategy. The Interaction Network Ontology (INO) includes a hierarchy of various interaction-related keywords useful for literature mining. Using VO, INO, and normalized E. coli gene names, we applied an ontology-based SciMiner literature mining strategy to mine all PubMed abstracts and retrieve E. coli vaccine-associated E. coli gene interactions. Four centrality metrics (i.e., degree, eigenvector, closeness, and betweenness) were calculated for identifying highly ranked genes and interaction types. Using vaccine-related PubMed abstracts, our study identified 11,350 sentences that contain 88 unique INO interactions types and 1,781 unique E. coli genes. Each sentence contained at least one interaction type and two unique E. coli genes. An E. coli gene interaction network of genes and INO interaction types was created. From this big network, a sub-network consisting of 5 E. coli vaccine genes, including carA, carB, fimH, fepA, and vat, and 62 other E. coli genes, and 25 INO interaction types was identified. While many interaction types represent direct interactions between two indicated genes, our study has also shown that many of these retrieved interaction types are indirect in that the two genes participated in the specified interaction process in a required but indirect process. Our centrality analysis of

Isolation and characterization of MMS-sensitive mutants of Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Prakash, L.; Prakash, S.

1977-01-01

We have isolated mutants sensitive to methyl methanesulfonate (MMS) in Saccharomyces cerevisiae. Alleles of rad1, rad4, rad6, rad52, rad55 and rad57 were found among these mms mutants. Twenty-nine of the mms mutants which complement the existing radiation-sensitive (rad and rev) mutants belong to 22 new complementation groups. Mutants from five complementation groups are sensitive only to MMS. Mutants of 11 complementation groups are sensitive to uv or x rays in addition to MMS, mutants of six complementation groups are sensitive to all three agents. The cross-sensitivities of these mms mutants to uv and x rays are discussed in terms of their possible involvement in DNA repair. Sporulation is reduced or absent in homozygous diploids of mms mutants from nine complementation groups

Digital technical map of Košice city

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Štefan Kuzevič

2007-06-01

Full Text Available The digital map of a city is comprised complex of the map. Process of creation of the digital map of a city is time consuming and financially demanding. The digital map is created as co-operation of the local authority, technology network administrator, companies, and local government. The exact and current digital map of the city utilizable for multilateral applications is the result of this co-operation. The digital map of city catch all important phenomenon and objects which are needed for making decisions and planning to all grades controlling the local authority. The Geographic information systems tools make possible updating digital map, analyses and syntheses spatial phenomena and theirs relationships. The digital map of city is created step by step. The digital technical map of city is one of very important part of the digital map. Illustration of the part digital technical map of Košice is showed in the Fig. 1.

Repair of UV-damaged incoming plasmid DNA in Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Keszenman-Pereyra, David

1990-01-01

A whole-cell transformation assay was used for the repair of UV-damaged plasma DNA in highly-transformable haploid strains of Saccharomyces cerevisiae having different repair capabilities. The experiments described demonstrate that three epistasis groups (Friedberg 1988) are involved in the repair of UV-incoming DNA and that the repair processes act less efficiently on incoming DNA than they do on chromosomal DNA. The implications of these findings for UV repair in Saccharomyces cerevisiae are discussed. (author)

Fatty acid metabolism in Saccharomyces cerevisiae

NARCIS (Netherlands)

van Roermund, C. W. T.; Waterham, H. R.; IJlst, L.; Wanders, R. J. A.

2003-01-01

Peroxisomes are essential subcellular organelles involved in a variety of metabolic processes. Their importance is underlined by the identification of a large group of inherited diseases in humans in which one or more of the peroxisomal functions are impaired. The yeast Saccharomyces cerevisiae has

Purification and biochemical characterisation of the yeast ABC transporter Pdr11p

DEFF Research Database (Denmark)

Laub, Katrine Rude

Sterols constitute an essential lipid class in eukaryotic membranes where intracellular distributions are highly regulated. In the yeast Saccharomyces cerevisiae sterol uptake has been attributed to the two plasma membrane-localised ATP-binding cassette (ABC) transporters, Aus1p and Pdr11p...... of the yeast ABC transporter Pdr11p. This includes optimising its overexpression utilising the galactose induction system in S. cerevisiae, screening for the best detergent to extract the protein from the membrane, and establishing purification and reconstitution protocols. By providing a purification...

Intracellular Signal Triggered by Cholera Toxin in Saccharomyces boulardii and Saccharomyces cerevisiae

Science.gov (United States)

Brandão, Rogelio L.; Castro, Ieso M.; Bambirra, Eduardo A.; Amaral, Sheila C.; Fietto, Luciano G.; Tropia, Maria José M.; Neves, Maria José; Dos Santos, Raquel G.; Gomes, Newton C. M.; Nicoli, Jacques R.

1998-01-01

As is the case for Saccharomyces boulardii, Saccharomyces cerevisiae W303 protects Fisher rats against cholera toxin (CT). The addition of glucose or dinitrophenol to cells of S. boulardii grown on a nonfermentable carbon source activated trehalase in a manner similar to that observed for S. cerevisiae. The addition of CT to the same cells also resulted in trehalase activation. Experiments performed separately on the A and B subunits of CT showed that both are necessary for activation. Similarly, the addition of CT but not of its separate subunits led to a cyclic AMP (cAMP) signal in both S. boulardii and S. cerevisiae. These data suggest that trehalase stimulation by CT probably occurred through the cAMP-mediated protein phosphorylation cascade. The requirement of CT subunit B for both the cAMP signal and trehalase activation indicates the presence of a specific receptor on the yeasts able to bind to the toxin, a situation similar to that observed for mammalian cells. This hypothesis was reinforced by experiments with 125I-labeled CT showing specific binding of the toxin to yeast cells. The adhesion of CT to a receptor on the yeast surface through the B subunit and internalization of the A subunit (necessary for the cAMP signal and trehalase activation) could be one more mechanism explaining protection against the toxin observed for rats treated with yeasts. PMID:9464394

Co-cultivation of non-conventional yeast with Saccharomyces cerevisiae to increase the aroma complexity of fermented beverages

NARCIS (Netherlands)

Rijswijck, van Irma M.H.

2017-01-01

Yeast are used as workhorses to convert hopped wort into beer. Conventionally, such yeasts belong to the genus Saccharomyces and most research on fermentation of wort for the production of beer has focussed on the species Saccharomyces cerevisiae and Saccharomyces