Sample records for strain background modifies

  1. Locating a modifier gene of Ovum mutant through crosses between DDK and C57BL/6J inbred strains in mice.

    PubMed

    Tan, Jing; Song, Gen Di; Song, Jia Sheng; Ren, Shi Hao; Li, Chun Li; Zheng, Zhen Yu; Zhao, Wei Dong

    2016-06-01

    A striking infertile phenotype has been discovered in the DDK strain of mouse. The DDK females are usually infertile when crossed with males of other inbred strains, whereas DDK males exhibit normal fertility in reciprocal crosses. This phenomenon is caused by mutation in the ovum (Om) locus on chromosome 11 and known as the DDK syndrome. Previously, some research groups reported that the embryonic mortality deviated from the semilethal rate in backcrosses between heterozygous (Om/+) females and males of other strains. This embryonic mortality exhibited an aggravated trend with increasing background genes of other strains. These results indicated that some modifier genes of Om were present in other strains. In the present study, a population of N₂2 (Om/+) females from the backcrosses between C57BL/6J (B6) and F₁ (B6♀ × DDK♂) was used to map potential modifier genes of Om. Quantitative trait locus showed that a major locus, namely Amom1 (aggravate modifier gene of Om 1), was located at the middle part of chromosome 9 in mice. The Amom1 could increase the expressivity of Om gene, thereby aggravating embryonic lethality when heterozygous (Om/+) females mated with males of B6 strain. Further, the 1.5 LOD-drop analysis indicated that the confidence interval was between 37.54 and 44.46 cM, ~6.92 cM. Amom1 is the first modifier gene of Om in the B6 background.

  2. Effect of genetic background on the dystrophic phenotype in mdx mice

    PubMed Central

    Coley, William D.; Bogdanik, Laurent; Vila, Maria Candida; Yu, Qing; Van Der Meulen, Jack H.; Rayavarapu, Sree; Novak, James S.; Nearing, Marie; Quinn, James L.; Saunders, Allison; Dolan, Connor; Andrews, Whitney; Lammert, Catherine; Austin, Andrew; Partridge, Terence A.; Cox, Gregory A.; Lutz, Cathleen; Nagaraju, Kanneboyina

    2016-01-01

    Genetic background significantly affects phenotype in multiple mouse models of human diseases, including muscular dystrophy. This phenotypic variability is partly attributed to genetic modifiers that regulate the disease process. Studies have demonstrated that introduction of the γ-sarcoglycan-null allele onto the DBA/2J background confers a more severe muscular dystrophy phenotype than the original strain, demonstrating the presence of genetic modifier loci in the DBA/2J background. To characterize the phenotype of dystrophin deficiency on the DBA/2J background, we created and phenotyped DBA/2J-congenic Dmdmdx mice (D2-mdx) and compared them with the original, C57BL/10ScSn-Dmdmdx (B10-mdx) model. These strains were compared with their respective control strains at multiple time points between 6 and 52 weeks of age. Skeletal and cardiac muscle function, inflammation, regeneration, histology and biochemistry were characterized. We found that D2-mdx mice showed significantly reduced skeletal muscle function as early as 7 weeks and reduced cardiac function by 28 weeks, suggesting that the disease phenotype is more severe than in B10-mdx mice. In addition, D2-mdx mice showed fewer central myonuclei and increased calcifications in the skeletal muscle, heart and diaphragm at 7 weeks, suggesting that their pathology is different from the B10-mdx mice. The new D2-mdx model with an earlier onset and more pronounced dystrophy phenotype may be useful for evaluating therapies that target cardiac and skeletal muscle function in dystrophin-deficient mice. Our data align the D2-mdx with Duchenne muscular dystrophy patients with the LTBP4 genetic modifier, making it one of the few instances of cross-species genetic modifiers of monogenic traits. PMID:26566673

  3. Absence of strong strain effects in behavioral analyses of Shank3-deficient mice

    PubMed Central

    Drapeau, Elodie; Dorr, Nate P.; Elder, Gregory A.; Buxbaum, Joseph D.

    2014-01-01

    Haploinsufficiency of SHANK3, caused by chromosomal abnormalities or mutations that disrupt one copy of the gene, leads to a neurodevelopmental syndrome called Phelan-McDermid syndrome, symptoms of which can include absent or delayed speech, intellectual disability, neurological changes and autism spectrum disorders. The SHANK3 protein forms a key structural part of the post-synaptic density. We previously generated and characterized mice with a targeted disruption of Shank3 in which exons coding for the ankyrin-repeat domain were deleted and expression of full-length Shank3 was disrupted. We documented specific deficits in synaptic function and plasticity, along with reduced reciprocal social interactions, in Shank3 heterozygous mice. Changes in phenotype owing to a mutation at a single locus are quite frequently modulated by other loci, most dramatically when the entire genetic background is changed. In mice, each strain of laboratory mouse represents a distinct genetic background and alterations in phenotype owing to gene knockout or transgenesis are frequently different across strains, which can lead to the identification of important modifier loci. We have investigated the effect of genetic background on phenotypes of Shank3 heterozygous, knockout and wild-type mice, using C57BL/6, 129SVE and FVB/Ntac strain backgrounds. We focused on observable behaviors with the goal of carrying out subsequent analyses to identify modifier loci. Surprisingly, there were very modest strain effects over a large battery of analyses. These results indicate that behavioral phenotypes associated with Shank3 haploinsufficiency are largely strain-independent. PMID:24652766

  4. Phenotypic detection of broad-spectrum beta-lactamases in microbiological practice

    PubMed Central

    Sedlakova, Miroslava Htoutou; Hanulik, Vojtech; Chroma, Magdalena; Hricova, Kristyna; Kolar, Milan; Latal, Tomas; Schaumann, Reiner; Rodloff, Arne C.

    2011-01-01

    Summary Background Enterobacteriaceae producing ESBL and AmpC enzymes can be associated with failure of antibiotic therapy and related morbidity and mortality. Their routine detection in microbiology laboratories is still a problem. The aim of this study was to compare the sensitivity of selected phenotypic methods. Material/Methods A total of 106 strains of the Enterobacteriaceae family were tested, in which molecular biology methods confirmed the presence of genes encoding ESBL or AmpC. In ESBL-positive strains, the sensitivity of the ESBL Etest (AB Biodisk) and a modified double-disk synergy test (DDST) were evaluated. AmpC strains were tested by a modified AmpC disk method using 3-aminophenylboronic acid. For simultaneous detection of ESBL and AmpC, the microdilution method with a modified set of antimicrobial agents was used. Results The sensitivity of the ESBL Etest was 95%; the modified DDST yielded 100% sensitivity for ESBL producers and the AmpC test correctly detected 95% of AmpC-positive strains. The sensitivity of the modified microdilution method was 87% and 95% for ESBL and AmpC beta lactamases, respectively. Conclusions The detection of ESBL and AmpC beta lactamases should be based on specific phenotypic methods such as the modified DDST, ESBL Etest, AmpC disk test and the modified microdilution method. PMID:21525803

  5. Effect of wine-based marinades on the behavior of Salmonella Typhimurium and background flora in beef fillets.

    PubMed

    Nisiotou, A; Chorianopoulos, N G; Gounadaki, A; Panagou, E Z; Nychas, G-J E

    2013-06-17

    The aim of this study was to evaluate the wine-based marinades to control the survival of acid-adapted and non-adapted Salmonella Typhimurium and background flora of fresh beef stored aerobically or under modified atmosphere. Beef slices were inoculated with a 3-strain cocktail of acid-adapted or non-adapted Salmonella Typhimurium strains DT 193, 4/74 and DSM 554 and marinated by immersion in wine (W) or wine supplemented with 0.3% thyme essential oil (WEO), for 12h at 4°C. Marinated slices were then stored under air or modified atmosphere conditions at 5°C. S. Typhimurium and background flora were followed for a 19-day period of storage. S. Typhimurium individual strains were monitored by pulsed field gel electrophoresis. Marination with wine significantly (P<0.05) reduced the background flora compared to the control (non-marinated). Furthermore, immersion of fillets in W or WEO marinades for 12h significantly (P<0.05) reduced the levels of S. Typhimurium compared to the non-marinated (control) samples by 1.1 and 1.4logCFU/g or 2.0 and 1.9logCFU/g for acid-adapted and non-adapted cells, respectively. Acid-adapted cells were more susceptible (P<0.05) to the addition of thyme essential oil in the wine marinade. The epidemic multi-drug resistant DT 193, the 4/74 and DSM 554 strains survived marination (for both W and WEO) and were detected at about similar proportions as revealed by PFGE results. Present results indicate that wine-based marinades are efficient, from a safety and shelf life stand point, in reducing pathogen's levels as well as the background beef flora. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Genetic Diversity Influences the Response of the Brain to Developmental Lead Exposure

    PubMed Central

    Schneider, Jay S.; Talsania, Keyur; Mettil, William; Anderson, David W.

    2014-01-01

    Although extrinsic factors, such as nutritional status, and some intrinsic genetic factors may modify susceptibility to developmental lead (Pb) poisoning, no studies have specifically examined the influence of genetic background on outcomes from Pb exposure. In this study, we used gene microarray profiling to identify Pb-responsive genes in rats of different genetic backgrounds, including inbred (Fischer 344 (F344)) and outbred (Long Evans (LE), Sprague Dawley (SD)) strains, to investigate the role that genetic variation may play in influencing outcomes from developmental Pb exposure. Male and female animals received either perinatal (gestation through lactation) or postnatal (birth through weaning) exposure to Pb in food (0, 250, or 750 ppm). RNA was extracted from the hippocampus at day 55 and hybridized to Affymetrix Rat Gene 1.0 ST Arrays. There were significant strain-specific effects of Pb on the hippocampal transcriptome with 978 transcripts differentially expressed in LE rats across all experimental groups, 269 transcripts differentially expressed in F344 rats, and only 179 transcripts differentially expressed in SD rats. These results were not due to strain-related differences in brain accumulation of Pb. Further, no genes were consistently differentially regulated in all experimental conditions. There was no set of “Pb toxicity” genes that are a molecular signature for Pb neurotoxicity that transcended sex, exposure condition, and strain. These results demonstrate the influence that strain and genetic background play in modifying the brain's response to developmental Pb exposure and may have relevance for better understanding the molecular underpinnings of the lack of a neurobehavioral signature in childhood Pb poisoning. PMID:24913800

  7. Engineered coryneform bacteria as a bio-tool for arsenic remediation.

    PubMed

    Villadangos, Almudena F; Ordóñez, Efrén; Pedre, Brandán; Messens, Joris; Gil, Jose A; Mateos, Luis M

    2014-12-01

    Despite current remediation efforts, arsenic contamination in water sources is still a major health problem, highlighting the need for new approaches. In this work, strains of the nonpathogenic and highly arsenic-resistant bacterium Corynebacterium glutamicum were used as inexpensive tools to accumulate inorganic arsenic, either as arsenate (As(V)) or arsenite (As(III)) species. The assays made use of "resting cells" from these strains, which were assessed under well-established conditions and compared with C. glutamicum background controls. The two mutant As(V)-accumulating strains were those used in a previously published study: (i) ArsC1/C2, in which the gene/s encoding the mycothiol-dependent arsenate reductases is/are disrupted, and (ii) MshA/C mutants unable to produce mycothiol, the low molecular weight thiol essential for arsenate reduction. The As(III)-accumulating strains were either those lacking the arsenite permease activities (Acr3-1 and Acr3-2) needed in As(III) release or recombinant strains overexpressing the aquaglyceroporin genes (glpF) from Corynebacterium diphtheriae or Streptomyces coelicolor, to improve As(III) uptake. Both genetically modified strains accumulated 30-fold more As(V) and 15-fold more As(III) than the controls. The arsenic resistance of the modified strains was inversely proportional to their metal accumulation ability. Our results provide the basis for investigations into the use of these modified C. glutamicum strains as a new bio-tool in arsenic remediation efforts.

  8. Strain Background Modifies Phenotypes in the ATP8B1-Deficient Mouse

    PubMed Central

    Vargas, Julie C.; Xu, Hongmei; Groen, Annamiek; Paulusma, Coen C.; Grenert, James P.; Pawlikowska, Ludmila; Sen, Saunak; Elferink, Ronald P. J. Oude; Bull, Laura N.

    2010-01-01

    Background Mutations in ATP8B1 (FIC1) underlie cases of cholestatic disease, ranging from chronic and progressive (progressive familial intrahepatic cholestasis) to intermittent (benign recurrent intrahepatic cholestasis). The ATP8B1-deficient mouse serves as an animal model of human ATP8B1 deficiency. Methodology/Principal Findings We investigated the effect of genetic background on phenotypes of ATP8B1-deficient and wild-type mice, using C57Bl/6 (B6), 129, and (B6-129) F1 strain backgrounds. B6 background resulted in greater abnormalities in ATP8B1-deficient mice than did 129 and/or F1 background. ATP8B1-deficient pups of B6 background gained less weight. In adult ATP8B1-deficient mice at baseline, those of B6 background had lower serum cholesterol levels, higher serum alkaline phosphatase levels, and larger livers. After challenge with cholate-supplemented diet, these mice exhibited higher serum alkaline phosphatase and bilirubin levels, greater weight loss and larger livers. ATP8B1-deficient phenotypes in mice of F1 and 129 backgrounds are usually similar, suggesting that susceptibility to manifestations of ATP8B1 deficiency may be recessive. We also detected differences in hepatobiliary phenotypes between wild-type mice of differing strains. Conclusions/Significance Our results indicate that the ATP8B1-deficient mouse in a B6 background may be a better model of human ATP8B1 deficiency and highlight the importance of informed background strain selection for mouse models of liver disease. PMID:20126555

  9. Genetic exchange of fimbrial alleles exemplifies the adaptive virulence strategy of Porphyromonas gingivalis.

    PubMed

    Kerr, Jennifer E; Abramian, Jared R; Dao, Doan-Hieu V; Rigney, Todd W; Fritz, Jamie; Pham, Tan; Gay, Isabel; Parthasarathy, Kavitha; Wang, Bing-yan; Zhang, Wenjian; Tribble, Gena D

    2014-01-01

    Porphyromonas gingivalis is a gram-negative anaerobic bacterium, a member of the human oral microbiome, and a proposed "keystone" pathogen in the development of chronic periodontitis, an inflammatory disease of the gingiva. P. gingivalis is a genetically diverse species, and is able to exchange chromosomal DNA between strains by natural competence and conjugation. In this study, we investigate the role of horizontal DNA transfer as an adaptive process to modify behavior, using the major fimbriae as our model system, due to their critical role in mediating interactions with the host environment. We show that P. gingivalis is able to exchange fimbrial allele types I and IV into four distinct strain backgrounds via natural competence. In all recombinants, we detected a complete exchange of the entire fimA allele, and the rate of exchange varies between the different strain backgrounds. In addition, gene exchange within other regions of the fimbrial genetic locus was identified. To measure the biological implications of these allele swaps we compared three genotypes of fimA in an isogenic background, strain ATCC 33277. We demonstrate that exchange of fimbrial allele type results in profound phenotypic changes, including the quantity of fimbriae elaborated, membrane blebbing, auto-aggregation and other virulence-associated phenotypes. Replacement of the type I allele with either the type III or IV allele resulted in increased invasion of gingival fibroblast cells relative to the isogenic parent strain. While genetic variability is known to impact host-microbiome interactions, this is the first study to quantitatively assess the adaptive effect of exchanging genes within the pan genome cloud. This is significant as it presents a potential mechanism by which opportunistic pathogens may acquire the traits necessary to modify host-microbial interactions.

  10. Core neuropathological abnormalities in progranulin-deficient mice are penetrant on multiple genetic backgrounds.

    PubMed

    Petkau, T L; Hill, A; Leavitt, B R

    2016-02-19

    Loss-of-function mutations in the progranulin gene (GRN) are a common cause of familial frontotemporal lobar degeneration (FTLD). A high degree of heterogeneity in the age-of-onset, duration of disease, and clinical presentation of FTLD, even among families carrying the same GRN mutation, suggests that additional modifying genes may be important to pathogenesis. Progranulin-knockout mice display subtle behavioral abnormalities and progressive neuropathological changes, as well as altered dendritic morphology and synaptic deficits in the hippocampus. In this study we evaluated multiple neuropathological endpoints in aged progranulin knockout mice and their wild-type littermates on two different genetic backgrounds: C57Bl/6 and 129/SvImJ. We find that in most brain regions, both strains are susceptible to progranulin-mediated neuropathological phenotypes, including astrogliosis, microgliosis, and highly accelerated deposition of the aging pigment lipofuscin. Neuroinflammation due to progranulin deficiency is exaggerated in the B6 strain and present, but less pronounced, in the 129 strain. Differences between the strains in hippocampal neuron counts and neuronal morphology suggest a complex role for progranulin in the hippocampus. We conclude that core progranulin-mediated neurodegenerative phenotypes are penetrant on multiple inbred mouse strains, but that genetic background modulates progranulin's role in neuroinflammation and hippocampal biology. Copyright © 2015 IBRO. Published by Elsevier Ltd. All rights reserved.

  11. Effects of strain and age on hepatic gene expression profiles in murine models of HFE-associated hereditary hemochromatosis.

    PubMed

    Lee, Seung-Min; Loguinov, Alexandre; Fleming, Robert E; Vulpe, Christopher D

    2015-01-01

    Hereditary hemochromatosis is an iron overload disorder most commonly caused by a defect in the HFE gene. While the genetic defect is highly prevalent, the majority of individuals do not develop clinically significant iron overload, suggesting the importance of genetic modifiers. Murine hfe knockout models have demonstrated that strain background has a strong effect on the severity of iron loading. We noted that hepatic iron loading in hfe-/- mice occurs primarily over the first postnatal weeks (loading phase) followed by a timeframe of relatively static iron concentrations (plateau phase). We thus evaluated the effects of background strain and of age on hepatic gene expression in Hfe knockout mice (hfe-/-). Hepatic gene expression profiles were examined using cDNA microarrays in 4- and 8-week-old hfe-/- and wild-type mice on two different genetic backgrounds, C57BL/6J (C57) and AKR/J (AKR). Genes differentially regulated in all hfe-/- mice groups, compared with wild-type mice, including those involved in cell survival, stress and damage responses and lipid metabolism. AKR strain-specific changes in lipid metabolism genes and C57 strain-specific changes in cell adhesion and extracellular matrix protein genes were detected in hfe-/- mice. Mouse strain and age are each significantly associated with hepatic gene expression profiles in hfe-/- mice. These affects may underlie or reflect differences in iron loading in these mice.

  12. Mismatch repair genes Mlh1 and Mlh3 modify CAG instability in Huntington's disease mice: genome-wide and candidate approaches.

    PubMed

    Pinto, Ricardo Mouro; Dragileva, Ella; Kirby, Andrew; Lloret, Alejandro; Lopez, Edith; St Claire, Jason; Panigrahi, Gagan B; Hou, Caixia; Holloway, Kim; Gillis, Tammy; Guide, Jolene R; Cohen, Paula E; Li, Guo-Min; Pearson, Christopher E; Daly, Mark J; Wheeler, Vanessa C

    2013-10-01

    The Huntington's disease gene (HTT) CAG repeat mutation undergoes somatic expansion that correlates with pathogenesis. Modifiers of somatic expansion may therefore provide routes for therapies targeting the underlying mutation, an approach that is likely applicable to other trinucleotide repeat diseases. Huntington's disease Hdh(Q111) mice exhibit higher levels of somatic HTT CAG expansion on a C57BL/6 genetic background (B6.Hdh(Q111) ) than on a 129 background (129.Hdh(Q111) ). Linkage mapping in (B6x129).Hdh(Q111) F2 intercross animals identified a single quantitative trait locus underlying the strain-specific difference in expansion in the striatum, implicating mismatch repair (MMR) gene Mlh1 as the most likely candidate modifier. Crossing B6.Hdh(Q111) mice onto an Mlh1 null background demonstrated that Mlh1 is essential for somatic CAG expansions and that it is an enhancer of nuclear huntingtin accumulation in striatal neurons. Hdh(Q111) somatic expansion was also abolished in mice deficient in the Mlh3 gene, implicating MutLγ (MLH1-MLH3) complex as a key driver of somatic expansion. Strikingly, Mlh1 and Mlh3 genes encoding MMR effector proteins were as critical to somatic expansion as Msh2 and Msh3 genes encoding DNA mismatch recognition complex MutSβ (MSH2-MSH3). The Mlh1 locus is highly polymorphic between B6 and 129 strains. While we were unable to detect any difference in base-base mismatch or short slipped-repeat repair activity between B6 and 129 MLH1 variants, repair efficiency was MLH1 dose-dependent. MLH1 mRNA and protein levels were significantly decreased in 129 mice compared to B6 mice, consistent with a dose-sensitive MLH1-dependent DNA repair mechanism underlying the somatic expansion difference between these strains. Together, these data identify Mlh1 and Mlh3 as novel critical genetic modifiers of HTT CAG instability, point to Mlh1 genetic variation as the likely source of the instability difference in B6 and 129 strains and suggest that MLH1 protein levels play an important role in driving of the efficiency of somatic expansions.

  13. Mismatch Repair Genes Mlh1 and Mlh3 Modify CAG Instability in Huntington's Disease Mice: Genome-Wide and Candidate Approaches

    PubMed Central

    Pinto, Ricardo Mouro; Dragileva, Ella; Kirby, Andrew; Lloret, Alejandro; Lopez, Edith; St. Claire, Jason; Panigrahi, Gagan B.; Hou, Caixia; Holloway, Kim; Gillis, Tammy; Guide, Jolene R.; Cohen, Paula E.; Li, Guo-Min; Pearson, Christopher E.; Daly, Mark J.; Wheeler, Vanessa C.

    2013-01-01

    The Huntington's disease gene (HTT) CAG repeat mutation undergoes somatic expansion that correlates with pathogenesis. Modifiers of somatic expansion may therefore provide routes for therapies targeting the underlying mutation, an approach that is likely applicable to other trinucleotide repeat diseases. Huntington's disease HdhQ111 mice exhibit higher levels of somatic HTT CAG expansion on a C57BL/6 genetic background (B6.HdhQ111) than on a 129 background (129.HdhQ111). Linkage mapping in (B6x129).HdhQ111 F2 intercross animals identified a single quantitative trait locus underlying the strain-specific difference in expansion in the striatum, implicating mismatch repair (MMR) gene Mlh1 as the most likely candidate modifier. Crossing B6.HdhQ111 mice onto an Mlh1 null background demonstrated that Mlh1 is essential for somatic CAG expansions and that it is an enhancer of nuclear huntingtin accumulation in striatal neurons. HdhQ111 somatic expansion was also abolished in mice deficient in the Mlh3 gene, implicating MutLγ (MLH1–MLH3) complex as a key driver of somatic expansion. Strikingly, Mlh1 and Mlh3 genes encoding MMR effector proteins were as critical to somatic expansion as Msh2 and Msh3 genes encoding DNA mismatch recognition complex MutSβ (MSH2–MSH3). The Mlh1 locus is highly polymorphic between B6 and 129 strains. While we were unable to detect any difference in base-base mismatch or short slipped-repeat repair activity between B6 and 129 MLH1 variants, repair efficiency was MLH1 dose-dependent. MLH1 mRNA and protein levels were significantly decreased in 129 mice compared to B6 mice, consistent with a dose-sensitive MLH1-dependent DNA repair mechanism underlying the somatic expansion difference between these strains. Together, these data identify Mlh1 and Mlh3 as novel critical genetic modifiers of HTT CAG instability, point to Mlh1 genetic variation as the likely source of the instability difference in B6 and 129 strains and suggest that MLH1 protein levels play an important role in driving of the efficiency of somatic expansions. PMID:24204323

  14. Identification of five novel modifier loci of ApcMin harbored in the BXH14 recombinant inbred strain

    PubMed Central

    Siracusa, Linda D.

    2012-01-01

    Every year thousands of people in the USA are diagnosed with small intestine and colorectal cancers (CRC). Although environmental factors affect disease etiology, uncovering underlying genetic factors is imperative for risk assessment and developing preventative therapies. Familial adenomatous polyposis is a heritable genetic disorder in which individuals carry germ-line mutations in the adenomatous polyposis coli (APC) gene that predisposes them to CRC. The Apc Min mouse model carries a point mutation in the Apc gene and develops polyps along the intestinal tract. Inbred strain background influences polyp phenotypes in Apc Min mice. Several Modifier of Min (Mom) loci that alter tumor phenotypes associated with the Apc Min mutation have been identified to date. We screened BXH recombinant inbred (RI) strains by crossing BXH RI females with C57BL/6J (B6) Apc Min males and quantitating tumor phenotypes in backcross progeny. We found that the BXH14 RI strain harbors five modifier loci that decrease polyp multiplicity. Furthermore, we show that resistance is determined by varying combinations of these modifier loci. Gene interaction network analysis shows that there are multiple networks with proven gene–gene interactions, which contain genes from all five modifier loci. We discuss the implications of this result for studies that define susceptibility loci, namely that multiple networks may be acting concurrently to alter tumor phenotypes. Thus, the significance of this work resides not only with the modifier loci we identified but also with the combinations of loci needed to get maximal protection against polyposis and the impact of this finding on human disease studies. Abbreviations:APCadenomatous polyposis coliGWASgenome-wide association studiesQTLquantitative trait lociSNPsingle-nucleotide polymorphism. PMID:22637734

  15. Background Nutrients Affect the Biotransformation of Tetracycline by Stenotrophomonas maltophilia as Revealed by Genomics and Proteomics.

    PubMed

    Leng, Yifei; Bao, Jianguo; Song, Dandan; Li, Jing; Ye, Mao; Li, Xu

    2017-09-19

    Certain bacteria are resistant to antibiotics and can even transform antibiotics in the environment. It is unclear how the molecular mechanisms underlying the resistance and biotransformation processes vary under different environmental conditions. The objective of this study is to investigate the molecular mechanisms of tetracycline resistance and biotransformation by Stenotrophomonas maltophilia strain DT1 under various background nutrient conditions. Strain DT1 was exposed to tetracycline for 7 days with four background nutrient conditions: no background (NB), peptone (P), peptone plus citrate (PC), and peptone plus glucose (PG). The biotransformation rate follows the order of PC > P > PG > NB ≈ 0. Genomic analysis showed that strain DT1 contained tet(X1), a gene encoding an FAD-binding monooxygenase, and eight peroxidase genes that could be relevant to tetracycline biotransformation. Quantitative proteomic analyses revealed that nodulation protein transported tetracycline outside of cells; hypoxanthine-guanine phosphoribosyltransferase facilitated the activation of the ribosomal protection proteins to prevent the binding of tetracycline to the ribosome and superoxide dismutase and peroxiredoxin-modified tetracycline molecules. Comparing different nutrient conditions showed that the biotransformation rates of tetracycline were positively correlated with the expression levels of superoxide dismutase.

  16. High Strain Rate Deformation Modeling of a Polymer Matrix Composite. Part 1; Matrix Constitutive Equations

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.; Stouffer, Donald C.

    1998-01-01

    Recently applications have exposed polymer matrix composite materials to very high strain rate loading conditions, requiring an ability to understand and predict the material behavior under these extreme conditions. In this first paper of a two part report, background information is presented, along with the constitutive equations which will be used to model the rate dependent nonlinear deformation response of the polymer matrix. Strain rate dependent inelastic constitutive models which were originally developed to model the viscoplastic deformation of metals have been adapted to model the nonlinear viscoelastic deformation of polymers. The modified equations were correlated by analyzing the tensile/ compressive response of both 977-2 toughened epoxy matrix and PEEK thermoplastic matrix over a variety of strain rates. For the cases examined, the modified constitutive equations appear to do an adequate job of modeling the polymer deformation response. A second follow-up paper will describe the implementation of the polymer deformation model into a composite micromechanical model, to allow for the modeling of the nonlinear, rate dependent deformation response of polymer matrix composites.

  17. Characterization of fatty acid modifying enzyme activity in staphylococcal mastitis isolates and other bacteria

    PubMed Central

    2012-01-01

    Background Fatty acid modifying enzyme (FAME) has been shown to modify free fatty acids to alleviate their bactericidal effect by esterifying fatty acids to cholesterol or alcohols. Although it has been shown in previous studies that FAME is required for Staphylococcus aureus survival in skin abscesses, FAME is poorly studied compared to other virulence factors. FAME activity had also been detected in coagulase-negative staphylococci (CNS). However, FAME activity was only surveyed after a bacterial culture was grown for 24 h. Therefore if FAME activity was earlier in the growth phase, it would not have been detected by the assay and those strains would have been labeled as FAME negative. Results Fifty CNS bovine mastitis isolates and several S. aureus, Escherichia coli, and Streptococcus uberis strains were assayed for FAME activity over 24 h. FAME activity was detected in 54% of CNS and 80% S. aureus strains surveyed but none in E. coli or S. uberis. While some CNS strains produced FAME activity comparable to the lab strain of S. aureus, the pattern of FAME activity varied among strains and across species of staphylococci. All CNS that produced FAME activity also exhibited lipase activity. Lipase activity relative to colony forming units of these CNS decreased over the 24 h growth period. No relationship was observed between somatic cell count in the milk and FAME activity in CNS. Conclusions Some staphylococcal species surveyed produced FAME activity, but E. coli and S. uberis strains did not. All FAME producing CNS exhibited lipase activity which may indicate that both these enzymes work in concert to alter fatty acids in the bacterial environment. PMID:22726316

  18. Genetic background influences age-related decline in visual and nonvisual retinal responses, circadian rhythms, and sleep☆

    PubMed Central

    Banks, Gareth; Heise, Ines; Starbuck, Becky; Osborne, Tamzin; Wisby, Laura; Potter, Paul; Jackson, Ian J.; Foster, Russell G.; Peirson, Stuart N.; Nolan, Patrick M.

    2015-01-01

    The circadian system is entrained to the environmental light/dark cycle via retinal photoreceptors and regulates numerous aspects of physiology and behavior, including sleep. These processes are all key factors in healthy aging showing a gradual decline with age. Despite their importance, the exact mechanisms underlying this decline are yet to be fully understood. One of the most effective tools we have to understand the genetic factors underlying these processes are genetically inbred mouse strains. The most commonly used reference mouse strain is C57BL/6J, but recently, resources such as the International Knockout Mouse Consortium have started producing large numbers of mouse mutant lines on a pure genetic background, C57BL/6N. Considering the substantial genetic diversity between mouse strains we expect there to be phenotypic differences, including differential effects of aging, in these and other strains. Such differences need to be characterized not only to establish how different mouse strains may model the aging process but also to understand how genetic background might modify age-related phenotypes. To ascertain the effects of aging on sleep/wake behavior, circadian rhythms, and light input and whether these effects are mouse strain-dependent, we have screened C57BL/6J, C57BL/6N, C3H-HeH, and C3H-Pde6b+ mouse strains at 5 ages throughout their life span. Our data show that sleep, circadian, and light input parameters are all disrupted by the aging process. Moreover, we have cataloged a number of strain-specific aging effects, including the rate of cataract development, decline in the pupillary light response, and changes in sleep fragmentation and the proportion of time spent asleep. PMID:25179226

  19. Genetic background influences age-related decline in visual and nonvisual retinal responses, circadian rhythms, and sleep.

    PubMed

    Banks, Gareth; Heise, Ines; Starbuck, Becky; Osborne, Tamzin; Wisby, Laura; Potter, Paul; Jackson, Ian J; Foster, Russell G; Peirson, Stuart N; Nolan, Patrick M

    2015-01-01

    The circadian system is entrained to the environmental light/dark cycle via retinal photoreceptors and regulates numerous aspects of physiology and behavior, including sleep. These processes are all key factors in healthy aging showing a gradual decline with age. Despite their importance, the exact mechanisms underlying this decline are yet to be fully understood. One of the most effective tools we have to understand the genetic factors underlying these processes are genetically inbred mouse strains. The most commonly used reference mouse strain is C57BL/6J, but recently, resources such as the International Knockout Mouse Consortium have started producing large numbers of mouse mutant lines on a pure genetic background, C57BL/6N. Considering the substantial genetic diversity between mouse strains we expect there to be phenotypic differences, including differential effects of aging, in these and other strains. Such differences need to be characterized not only to establish how different mouse strains may model the aging process but also to understand how genetic background might modify age-related phenotypes. To ascertain the effects of aging on sleep/wake behavior, circadian rhythms, and light input and whether these effects are mouse strain-dependent, we have screened C57BL/6J, C57BL/6N, C3H-HeH, and C3H-Pde6b+ mouse strains at 5 ages throughout their life span. Our data show that sleep, circadian, and light input parameters are all disrupted by the aging process. Moreover, we have cataloged a number of strain-specific aging effects, including the rate of cataract development, decline in the pupillary light response, and changes in sleep fragmentation and the proportion of time spent asleep. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  20. Genetic Background Has a Major Impact on Differences in Sleep Resulting from Environmental Influences in Drosophila

    PubMed Central

    Zimmerman, John E.; Chan, May T.; Jackson, Nicholas; Maislin, Greg; Pack, Allan I.

    2012-01-01

    Study Objectives: To determine the effect of different genetic backgrounds on demographic and environmental interventions that affect sleep and evaluate variance of these measures; and to evaluate sleep and variance of sleep behaviors in 6 divergent laboratory strains of common origin. Design: Assessment of the effects of age, sex, mating status, food sources, and social experience using video analysis of sleep behavior in 2 different strains of Drosophila, white1118ex (w1118ex) and white Canton-S (wCS10). Sleep was also determined for 6 laboratory strains of Canton-S and 3 inbred lines. The variance of total sleep was determined for all groups and conditions. Measurements and Results: The circadian periods and the effects of age upon sleep were the same between w1118ex and wCS10 strains. However, the w1118ex and wCS10 strains demonstrated genotype-dependent differences in the effects upon sleep of sex, mating status, social experience, and being on different foods. Variance of total sleep was found to differ in a genotype dependent manner for interventions between the w1118ex and wCS10 strains. Six different laboratory Canton-S strains were found to have significantly different circadian periods (P < 0.001) and sleep phenotypes (P < 0.001). Three inbred lines showed reduced variance for sleep measurements. Conclusions: One must control environmental conditions in a rigorously consistent manner to ensure that sleep data may be compared between experiments. Genetic background has a significant impact upon changes in sleep behavior and variance of behavior due to demographic factors and environmental interventions. This represents an opportunity to discover new genes that modify sleep/wake behavior. Citation: Zimmerman JE; Chan MT; Jackson N; Maislin G; Pack AI. Genetic background has a major impact on differences in sleep resulting from environmental influences in Drosophila. SLEEP 2012;35(4):545-557. PMID:22467993

  1. A novel modified-indirect ELISA based on spherical body protein 4 for detecting antibody during acute and long-term infections with diverse babesia bovis strains

    USDA-ARS?s Scientific Manuscript database

    Background: Cattle persistently infected with Babesia bovis are reservoirs for intra- and inter-herd transmission. Since B. bovis is considered a persistent infection, developing a reliable, high-throughput assay that detects antibody during all stages of the infection could be pivotal for establish...

  2. MSH3 polymorphisms and protein levels affect CAG repeat instability in Huntington's disease mice.

    PubMed

    Tomé, Stéphanie; Manley, Kevin; Simard, Jodie P; Clark, Greg W; Slean, Meghan M; Swami, Meera; Shelbourne, Peggy F; Tillier, Elisabeth R M; Monckton, Darren G; Messer, Anne; Pearson, Christopher E

    2013-01-01

    Expansions of trinucleotide CAG/CTG repeats in somatic tissues are thought to contribute to ongoing disease progression through an affected individual's life with Huntington's disease or myotonic dystrophy. Broad ranges of repeat instability arise between individuals with expanded repeats, suggesting the existence of modifiers of repeat instability. Mice with expanded CAG/CTG repeats show variable levels of instability depending upon mouse strain. However, to date the genetic modifiers underlying these differences have not been identified. We show that in liver and striatum the R6/1 Huntington's disease (HD) (CAG)∼100 transgene, when present in a congenic C57BL/6J (B6) background, incurred expansion-biased repeat mutations, whereas the repeat was stable in a congenic BALB/cByJ (CBy) background. Reciprocal congenic mice revealed the Msh3 gene as the determinant for the differences in repeat instability. Expansion bias was observed in congenic mice homozygous for the B6 Msh3 gene on a CBy background, while the CAG tract was stabilized in congenics homozygous for the CBy Msh3 gene on a B6 background. The CAG stabilization was as dramatic as genetic deficiency of Msh2. The B6 and CBy Msh3 genes had identical promoters but differed in coding regions and showed strikingly different protein levels. B6 MSH3 variant protein is highly expressed and associated with CAG expansions, while the CBy MSH3 variant protein is expressed at barely detectable levels, associating with CAG stability. The DHFR protein, which is divergently transcribed from a promoter shared by the Msh3 gene, did not show varied levels between mouse strains. Thus, naturally occurring MSH3 protein polymorphisms are modifiers of CAG repeat instability, likely through variable MSH3 protein stability. Since evidence supports that somatic CAG instability is a modifier and predictor of disease, our data are consistent with the hypothesis that variable levels of CAG instability associated with polymorphisms of DNA repair genes may have prognostic implications for various repeat-associated diseases.

  3. MSH3 Polymorphisms and Protein Levels Affect CAG Repeat Instability in Huntington's Disease Mice

    PubMed Central

    Simard, Jodie P.; Clark, Greg W.; Slean, Meghan M.; Swami, Meera; Shelbourne, Peggy F.; Tillier, Elisabeth R. M.; Monckton, Darren G.; Messer, Anne; Pearson, Christopher E.

    2013-01-01

    Expansions of trinucleotide CAG/CTG repeats in somatic tissues are thought to contribute to ongoing disease progression through an affected individual's life with Huntington's disease or myotonic dystrophy. Broad ranges of repeat instability arise between individuals with expanded repeats, suggesting the existence of modifiers of repeat instability. Mice with expanded CAG/CTG repeats show variable levels of instability depending upon mouse strain. However, to date the genetic modifiers underlying these differences have not been identified. We show that in liver and striatum the R6/1 Huntington's disease (HD) (CAG)∼100 transgene, when present in a congenic C57BL/6J (B6) background, incurred expansion-biased repeat mutations, whereas the repeat was stable in a congenic BALB/cByJ (CBy) background. Reciprocal congenic mice revealed the Msh3 gene as the determinant for the differences in repeat instability. Expansion bias was observed in congenic mice homozygous for the B6 Msh3 gene on a CBy background, while the CAG tract was stabilized in congenics homozygous for the CBy Msh3 gene on a B6 background. The CAG stabilization was as dramatic as genetic deficiency of Msh2. The B6 and CBy Msh3 genes had identical promoters but differed in coding regions and showed strikingly different protein levels. B6 MSH3 variant protein is highly expressed and associated with CAG expansions, while the CBy MSH3 variant protein is expressed at barely detectable levels, associating with CAG stability. The DHFR protein, which is divergently transcribed from a promoter shared by the Msh3 gene, did not show varied levels between mouse strains. Thus, naturally occurring MSH3 protein polymorphisms are modifiers of CAG repeat instability, likely through variable MSH3 protein stability. Since evidence supports that somatic CAG instability is a modifier and predictor of disease, our data are consistent with the hypothesis that variable levels of CAG instability associated with polymorphisms of DNA repair genes may have prognostic implications for various repeat-associated diseases. PMID:23468640

  4. Is the association between high strain work and depressive symptoms modified by private life social support: a cohort study of 1,074 Danish employees?

    PubMed Central

    2014-01-01

    Background Previous studies have shown that psychosocial working conditions characterized by high psychological demands and low decision latitude (i.e., high strain work) are associated with increased risk of depressive symptoms. Little is known, however, concerning how this association may be modified by factors outside the working environment. This article examines the modifying role of private life social support in the relation between high strain work and the development of severe depressive symptoms. Methods Data were questionnaire-based, collected from a cross-occupational sample of 1,074 Danish employees. At baseline, all participants were free of severe depressive symptoms, measured by the Mental Health Inventory. High strain work was defined by the combination of high psychological demands at work and low control, measured with multi-dimensional scales. Private life social support was operationalized as the number of life domains with confidants and dichotomized as low (0–1 domains) or high (2 or more domains). Using logistic regression we examined the risk of onset of severe depressive symptoms, adjusting for sex, age, occupational position, and prior depressive symptoms. Results Separately, neither high strain work nor low private life social support statistically significantly predicted depressive symptoms. However, participants with joint exposure to high strain work and low private life social support had an Odds ratio (OR) for severe depressive symptoms of 3.41 (95% CI: 1.36-8.58), compared to participants with no work strain and high private life social support. There was no increased risk for participants with high strain work and high private life social support (OR = 1.32, 95% CI: 0.65-2.68). The interaction term for departure from additivity was, however, not statistically significant (p = 0.18). Conclusions Our findings suggest that high strain work may increase risk of depressive symptoms in individuals with low private life social support, although the effect-modification was statistically non-significant. Larger studies are needed to further establish the role of private life social support in the relation between high strain work and depression. PMID:25005843

  5. Open Field Release of Genetically Engineered Sterile Male Aedes aegypti in Malaysia

    PubMed Central

    Raduan, Norzahira; Kwee Wee, Lim; Hong Ming, Wong; Guat Ney, Teoh; Rahidah A.A., Siti; Salman, Sawaluddin; Subramaniam, Selvi; Nordin, Oreenaiza; Hanum A.T., Norhaida; Angamuthu, Chandru; Marlina Mansor, Suria; Lees, Rosemary S.; Naish, Neil; Scaife, Sarah; Gray, Pam; Labbé, Geneviève; Beech, Camilla; Nimmo, Derric; Alphey, Luke; Vasan, Seshadri S.; Han Lim, Lee; Wasi A., Nazni; Murad, Shahnaz

    2012-01-01

    Background Dengue is the most important mosquito-borne viral disease. In the absence of specific drugs or vaccines, control focuses on suppressing the principal mosquito vector, Aedes aegypti, yet current methods have not proven adequate to control the disease. New methods are therefore urgently needed, for example genetics-based sterile-male-release methods. However, this requires that lab-reared, modified mosquitoes be able to survive and disperse adequately in the field. Methodology/Principal Findings Adult male mosquitoes were released into an uninhabited forested area of Pahang, Malaysia. Their survival and dispersal was assessed by use of a network of traps. Two strains were used, an engineered ‘genetically sterile’ (OX513A) and a wild-type laboratory strain, to give both absolute and relative data about the performance of the modified mosquitoes. The two strains had similar maximum dispersal distances (220 m), but mean distance travelled of the OX513A strain was lower (52 vs. 100 m). Life expectancy was similar (2.0 vs. 2.2 days). Recapture rates were high for both strains, possibly because of the uninhabited nature of the site. Conclusions/Significance After extensive contained studies and regulatory scrutiny, a field release of engineered mosquitoes was safely and successfully conducted in Malaysia. The engineered strain showed similar field longevity to an unmodified counterpart, though in this setting dispersal was reduced relative to the unmodified strain. These data are encouraging for the future testing and implementation of genetic control strategies and will help guide future field use of this and other engineered strains. PMID:22970102

  6. Development of an optically-based tension-indicating implanted orthopedic screw with a luminescent spectral ruler

    NASA Astrophysics Data System (ADS)

    Ravikumar, Nakul; Rogalski, Melissa M.; Benza, Donny; Lake, Joshua; Urban, Matthew; Pelham, Hunter; Anker, Jeffrey N.; DesJardins, John D.

    2017-03-01

    An orthopaedic screw was designed with an optical tension-indicator to non-invasively quantify screw tension and monitor the load sharing between the bone and the implant. The screw both applies load to the bone, and measures this load by reporting the strain on the screw. The screw contains a colorimetric optical encoder that converts axial strain into colorimetric changes visible through the head of the screw, or luminescent spectral changes that are detected through tissue. Screws were tested under cyclic mechanical loading to mimic in-vivo conditions to verify the sensitivity, repeatability, and reproducibility of the sensor. In the absence to tissue, color was measured using a digital camera as a function of axial load on a stainless steel cannulated (hollow) orthopedic screw, modified by adding a passive colorimetric strain gauge through the central hole. The sensor was able to quantify clinically-relevant bone healing strains. The sensor exhibited good repeatability and reproducibility but also displayed hysteresis due to the internal mechanics of the screw. The strain indicator was also modified for measurement through tissue by replacing the reflective colorimetric sensor with a low-background X-ray excited optical luminescence signal. Luminescent spectra were acquired through 6 mm of chicken breast tissue. Overall, this research shows feasibility for a unique device which quantifies the strain on an orthopedic screw. Future research will involve reducing hysteresis by changing the mechanism of strain transduction in the screw, miniaturizing the luminescent strain gauge, monitoring bending as well as tension, using alternative luminescent spectral rulers based upon near infrared fluorescence or upconversion luminescence, and application to monitoring changes in pretension and load sharing during bone healing.

  7. Parallel universes of Black Six biology.

    PubMed

    Kraev, Alexander

    2014-07-19

    Creation of lethal and synthetic lethal mutations in an experimental organism is a cornerstone of genetic dissection of gene function, and is related to the concept of an essential gene. Common inbred mouse strains carry background mutations, which can act as genetic modifiers, interfering with the assignment of gene essentiality. The inbred strain C57BL/6J, commonly known as "Black Six", stands out, as it carries a spontaneous homozygous deletion in the nicotinamide nucleotide transhydrogenase (Nnt) gene [GenBank: AH009385.2], resulting in impairment of steroidogenic mitochondria of the adrenal gland, and a multitude of indirect modifier effects, coming from alteration of glucocorticoid-regulated processes. Over time, the popular strain has been used, by means of gene targeting technology, to assign "essential" and "redundant" qualifiers to numerous genes, thus creating an internally consistent "parallel universe" of knowledge. It is unrealistic to suggest phasing-out of this strain, given the scope of shared resources built around it, however, continuing on the road of "strain-unawareness" will result in profound waste of effort, particularly where translational research is concerned. The review analyzes the historical roots of this phenomenon and proposes that building of "parallel universes" should be urgently made visible to a critical reader by obligatory use of unambiguous and persistent tags in publications and databases, such as hypertext links, pointing to a vendor's strain description web page, or to a digital object identifier (d.o.i.) of the original publication, so that any research done exclusively in C57BL/6J, could be easily identified. This article was reviewed by Dr. Neil Smalheiser and Dr. Miguel Andrade-Navarro.

  8. Application of synthetic biology in cyanobacteria and algae

    PubMed Central

    Wang, Bo; Wang, Jiangxin; Zhang, Weiwen; Meldrum, Deirdre R.

    2012-01-01

    Cyanobacteria and algae are becoming increasingly attractive cell factories for producing renewable biofuels and chemicals due to their ability to capture solar energy and CO2 and their relatively simple genetic background for genetic manipulation. Increasing research efforts from the synthetic biology approach have been made in recent years to modify cyanobacteria and algae for various biotechnological applications. In this article, we critically review recent progresses in developing genetic tools for characterizing or manipulating cyanobacteria and algae, the applications of genetically modified strains for synthesizing renewable products such as biofuels and chemicals. In addition, the emergent challenges in the development and application of synthetic biology for cyanobacteria and algae are also discussed. PMID:23049529

  9. Chemical genomic guided engineering of gamma-valerolactone tolerant yeast.

    PubMed

    Bottoms, Scott; Dickinson, Quinn; McGee, Mick; Hinchman, Li; Higbee, Alan; Hebert, Alex; Serate, Jose; Xie, Dan; Zhang, Yaoping; Coon, Joshua J; Myers, Chad L; Landick, Robert; Piotrowski, Jeff S

    2018-01-12

    Gamma valerolactone (GVL) treatment of lignocellulosic bomass is a promising technology for degradation of biomass for biofuel production; however, GVL is toxic to fermentative microbes. Using a combination of chemical genomics with the yeast (Saccharomyces cerevisiae) deletion collection to identify sensitive and resistant mutants, and chemical proteomics to monitor protein abundance in the presence of GVL, we sought to understand the mechanism toxicity and resistance to GVL with the goal of engineering a GVL-tolerant, xylose-fermenting yeast. Chemical genomic profiling of GVL predicted that this chemical affects membranes and membrane-bound processes. We show that GVL causes rapid, dose-dependent cell permeability, and is synergistic with ethanol. Chemical genomic profiling of GVL revealed that deletion of the functionally related enzymes Pad1p and Fdc1p, which act together to decarboxylate cinnamic acid and its derivatives to vinyl forms, increases yeast tolerance to GVL. Further, overexpression of Pad1p sensitizes cells to GVL toxicity. To improve GVL tolerance, we deleted PAD1 and FDC1 in a xylose-fermenting yeast strain. The modified strain exhibited increased anaerobic growth, sugar utilization, and ethanol production in synthetic hydrolysate with 1.5% GVL, and under other conditions. Chemical proteomic profiling of the engineered strain revealed that enzymes involved in ergosterol biosynthesis were more abundant in the presence of GVL compared to the background strain. The engineered GVL strain contained greater amounts of ergosterol than the background strain. We found that GVL exerts toxicity to yeast by compromising cellular membranes, and that this toxicity is synergistic with ethanol. Deletion of PAD1 and FDC1 conferred GVL resistance to a xylose-fermenting yeast strain by increasing ergosterol accumulation in aerobically grown cells. The GVL-tolerant strain fermented sugars in the presence of GVL levels that were inhibitory to the unmodified strain. This strain represents a xylose fermenting yeast specifically tailored to GVL produced hydrolysates.

  10. Quantitative trait loci that control body weight and obesity in an F2 intercross between C57BL/6J and DDD.Cg-Ay mice.

    PubMed

    Suto, Jun-ichi

    2011-07-01

    I have developed a congenic mouse strain for the A(y) allele at the agouti locus in an inbred DDD/Sgn strain, DDD.Cg-A(y). DDD.Cg-A(y) females are extremely obese and significantly heavier than B6.Cg-A(y) females. The objectives of this study were to determine the genetic basis of obesity in DDD.Cg-A(y) mice, and to determine whether or not their high body weight was due to the presence of DDD background-specific modifiers. I performed quantitative trait locus (QTL) analyses for body weight and body mass index in two types of F(2) mice [F2 A(y) (F(2) mice carrying the A(y) allele) and F(2) non-A(y) (F2 mice without the A(y) allele)] produced by crossing C57BL/6J females and DDD.Cg-A(y) males. The results of the QTL analysis of F(2) A(y) mice were very similar to those obtained for F(2) non-A(y) mice. It was unlikely that the high body weight of DDD.Cg-A(y) mice was due to the presence of specific modifiers. When both F(2) datasets were merged and analyzed, four significant body weight QTLs were identified on chromosomes 6, 9, and 17 (2 loci) and four significant obesity QTLs were identified on chromosomes 1, 6, 9, and 17. Although the presence of DDD background-specific modifiers was not confirmed, a multifactorial basis of obesity in DDD.Cg-A(y) females was thus revealed.

  11. Geobacteraceae strains and methods

    DOEpatents

    Lovley, Derek R.; Nevin, Kelly P.; Yi, Hana

    2015-07-07

    Embodiments of the present invention provide a method of producing genetically modified strains of electricigenic microbes that are specifically adapted for the production of electrical current in microbial fuel cells, as well as strains produced by such methods and fuel cells using such strains. In preferred embodiments, the present invention provides genetically modified strains of Geobacter sulfurreducens and methods of using such strains.

  12. Antibiotic-modifying activity of riachin, a non-cyanogenic cyanoglycoside extracted from Bauhinia pentandra

    PubMed Central

    de Farias, Pablo Antonio Maia; Figueredo, Fernando Gomes; Lucas, Aline Maria Brito; de Moura, Rafael Barbosa; Coutinho, Henrique Douglas Melo; da Silva, Tania Maria Sarmento; Martin, Ana Luiza de Aguiar Rocha; Fonteles, Marta Maria de França

    2015-01-01

    Background The search for new active compounds from the Brazilian flora has intensified in recent years, especially for new drugs with antibiotic potential. Accordingly, the aim of this study was to determine whether riachin has antibiotic activity in itself or is able to modulate the activity of conventional antibiotics. Methods A non-cyanogenic cyanoglycoside known as riachin was isolated from Bauhinia pentandra, and was tested alone and in combination with three antibiotics (clindamycin, amikacin, and gentamicin) against multiresistant bacterial strains (Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus). Results Riachin did not show significant antibiotic activity when tested alone against any strain (P>0.05). However, when combined with conventional antibiotics, it showed drug-modifying activity against strains of S. aureus exposed to clindamycin (P<0.001) as well as against P. aeruginosa exposed to amikacin (P<0.001). Although riachin did not show direct antibiotic activity, it had synergistic activity when combined with amikacin or clindamycin. The mechanism of action of this synergism is under investigation. Conclusion The results of this work demonstrate that some substances of natural origin can enhance the effectiveness of certain antibiotics, which means a substantial reduction in the drug dose required and possibly in consequent adverse events for patients. PMID:26109849

  13. Modification of Salmonella Typhimurium Motility by the Probiotic Yeast Strain Saccharomyces boulardii

    PubMed Central

    Pontier-Bres, Rodolphe; Prodon, François; Munro, Patrick; Rampal, Patrick; Lemichez, Emmanuel; Peyron, Jean François; Czerucka, Dorota

    2012-01-01

    Background Motility is an important component of Salmonella enterica serovar Typhimurium (ST) pathogenesis allowing the bacteria to move into appropriate niches, across the mucus layer and invade the intestinal epithelium. In vitro, flagellum-associated motility is closely related to the invasive properties of ST. The probiotic yeast Saccharomyces boulardii BIOCODEX (S.b-B) is widely prescribed for the prophylaxis and treatment of diarrheal diseases caused by bacteria or antibiotics. In case of Salmonella infection, S.b-B has been shown to decrease ST invasion of T84 colon cell line. The present study was designed to investigate the impact of S.b-B on ST motility. Methodology/Principal Findings Experiments were performed on human colonic T84 cells infected by the Salmonella strain 1344 alone or in the presence of S.b-B. The motility of Salmonella was recorded by time-lapse video microscopy. Next, a manual tracking was performed to analyze bacteria dynamics (MTrackJ plugin, NIH image J software). This revealed that the speed of bacterial movement was modified in the presence of S.b-B. The median curvilinear velocity (CLV) of Salmonella incubated alone with T84 decreased from 43.3 µm/sec to 31.2 µm/sec in the presence of S.b-B. Measurement of track linearity (TL) showed similar trends: S.b-B decreased by 15% the number of bacteria with linear tract (LT) and increased by 22% the number of bacteria with rotator tract (RT). Correlation between ST motility and invasion was further established by studying a non-motile flagella-deficient ST strain. Indeed this strain that moved with a CLV of 0.5 µm/sec, presented a majority of RT and a significant decrease in invasion properties. Importantly, we show that S.b-B modified the motility of the pathogenic strain SL1344 and significantly decreased invasion of T84 cells by this strain. Conclusions This study reveals that S.b-B modifies Salmonella's motility and trajectory which may account for the modification of Salmonella's invasion. PMID:22442723

  14. Mouse Sperm Cryopreservation and Recovery of Genetically Modified Mice.

    PubMed

    Low, Benjamin E; Taft, Rob A; Wiles, Michael V

    2016-01-01

    Highly definable genetically, the humble mouse is the "reagent" mammal of choice with which to probe and begin to understand the human condition in all its complexities. With the recent advance in direct genome editing via targeted nucleases, e.g., TALEN and CRISPR/Cas9, the possibilities in using these sophisticated tools have increased substantially leading to a massive increase in the variety of strain numbers of genetically modified lines. With this increase comes a greater need to economically and creatively manage their numbers. Further, once characterized, lines may be of limited use but still need to be archived in a format allowing their rapid resurrection. Further, maintaining colonies on "the shelf" is financially draining and carries potential risks including natural disaster loss, disease, and strain contamination. Here we outline a simple and economic protocol to cryopreserve mouse sperm from many different genetic backgrounds, and outline its recovery via in vitro fertilization (IVF). The combined use of sperm cryopreservation and IVF now allows a freedom and versatility in mouse management facilitating rapid line close down with the option to later recover and rapidly expand as needed.

  15. Is there a role for modified probiotics as beneficial microbes: a systematic review of the literature.

    PubMed

    Zorzela, L; Ardestani, S K; McFarland, L V; Vohra, S

    2017-10-13

    Our objective was to conduct a systematic review and meta-analysis for the use of modified (heat-killed or sonicated) probiotics for the efficacy and safety to prevent and treat various diseases. Recent clinical research has focused on living strains of probiotics, but use in high-risk patients and potential adverse reactions including bacteremia has focused interest on alternatives to the use of live probiotics. We searched MEDLINE/PubMed, Embase, Cochrane Central Register of Controlled Trials, CINAHL, Alt Health Watch, Web of Science, Scopus, PubMed, from inception to February 14, 2017 for randomised controlled trials involving modified probiotic strains. The primary outcome was efficacy to prevent or treat disease and the secondary outcome was incidence of adverse events. A total of 40 trials were included (n=3,913): 14 trials (15 arms with modified probiotics and 20 control arms) for the prevention of diseases and 26 trials (29 arms with modified probiotics and 32 control arms) for treatment of various diseases. Modified microbes were compared to either placebo (44%), or the same living probiotic strain (39%) or to only standard therapies (17%). Modified microbes were not significantly more or less effective than the living probiotic in 86% of the preventive trials and 69% of the treatment trials. Modified probiotic strains were significantly more effective in 15% of the treatment trials. Incidence rates of adverse events were similar for modified and living probiotics and other control groups, but many trials did not collect adequate safety data. Although several types of modified probiotics showed significant efficacy over living strains of probiotics, firm conclusions could not be reached due to the limited number of trials using the same type of modified microbe (strain, daily dose and duration) for a specific disease indication. Further research may illuminate other strains of modified probiotics that may have potential as clinical biotherapeutics.

  16. Dynamic strain aging behavior of modified 9Cr-1Mo and reduced activation ferritic martensitic steels under low cycle fatigue

    NASA Astrophysics Data System (ADS)

    Mariappan, K.; Shankar, Vani; Sandhya, R.; Prasad Reddy, G. V.; Mathew, M. D.

    2013-04-01

    Influence of temperature and strain rate on low cycle fatigue (LCF) behavior of modified 9Cr-1Mo ferritic martensitic steel and 1.4W-0.06Ta reduced activation ferritic martensitic (RAFM) steel in normalized and tempered conditions was studied. Total strain controlled LCF tests between 300 and 873 K on modified 9Cr-1Mo steel and RAFM steel and at various strain rates on modified 9Cr-1Mo steel were performed at total strain amplitude of ±0.6%. Both the steels showed continuous cyclic softening at all temperatures. Whereas manifestations of dynamic strain aging (DSA) were observed in both the steels which decreased fatigue life at intermediate temperatures, at higher temperatures, oxidation played a crucial role in decreasing fatigue life.

  17. Instability of the insertional mutation in CftrTgH(neoim)Hgu cystic fibrosis mouse model

    PubMed Central

    Charizopoulou, Nikoletta; Jansen, Silke; Dorsch, Martina; Stanke, Frauke; Dorin, Julia R; Hedrich, Hans-Jürgen; Tümmler, Burkhard

    2004-01-01

    Background A major boost to the cystic fibrosis disease research was given by the generation of various mouse models using gene targeting in embryonal stem cells. Moreover, the introduction of the same mutation on different inbred strains generating congenic strains facilitated the search for modifier genes. From the original CftrTgH(neoim)Hgu CF mouse model we have generated using strict brother × sister mating two inbred CftrTgH(neoim)Hgu mouse lines (CF/1 and CF/3). Thereafter, the insertional mutation was introgressed from CF/3 into three inbred backgrounds (C57BL/6, BALB/c, DBA/2J) generating congenic animals. In every backcross cycle germline transmission of the insertional mutation was monitored by direct probing the insertion via Southern RFLP. In order to bypass this time consuming procedure we devised an alternative PCR based protocol whereby mouse strains are differentiated at the Cftr locus by Cftr intragenic microsatellite genotypes that are tightly linked to the disrupted locus. Results Using this method we were able to identify animals carrying the insertional mutation based upon the differential haplotypic backgrounds of the three inbred strains and the mutant CftrTgH(neoim)Hgu at the Cftr locus. Moreover, this method facilitated the identification of the precise vector excision from the disrupted Cftr locus in two out of 57 typed animals. This reversion to wild type status took place without any loss of sequence revealing the instability of insertional mutations during the production of congenic animals. Conclusions We present intragenic microsatellite markers as a tool for fast and efficient identification of the introgressed locus of interest in the recipient strain during congenic animal breeding. Moreover, the same genotyping method allowed the identification of a vector excision event, posing questions on the stability of insertional mutations in mice. PMID:15102331

  18. Modified Newcastle disease virus vectors expressing the H5 hemagglutinin induce enhanced protection against highly pathogenic H5N1 avian influenza virus in chickens

    PubMed Central

    Kim, Shin-Hee; Paldurai, Anandan; Xiao, Sa; Collins, Peter L.; Samal, Siba K.

    2016-01-01

    Naturally-occurring attenuated strains of Newcastle disease virus (NDV) are being developed as vaccine vectors for use in poultry and humans. However, some NDV strains, such as Beaudette C (BC), may retain too much virulence in poultry for safe use, and more highly attenuated strains may be suboptimally immunogenic. We therefore modified the BC strain by changing the multibasic cleavage site sequence of the F protein to the dibasic sequence of avirulent strain LaSota. Additionally, the BC, F, and HN proteins were modified in several ways to enhance virus replication. These modified BC-derived vectors and the LaSota strain were engineered to express the hemagglutin (HA) protein of H5N1 highly pathogenic influenza virus (HPAIV). In general, the modified BC-based vectors expressing HA replicated better than LaSota/HA, and expressed higher levels of HA protein. Pathogenicity tests indicated that all the modified viruses were highly attenuated in chickens. Based on in vitro characterization, two of the modified BC vectors were chosen for evaluation in chickens as vaccine vectors against H5N1 HPAIV A/Vietnam/1203/04. Immunization of chickens with rNDV vector vaccines followed by challenge with HPAIV demonstrated high levels of protection against clinical disease and mortality. However, only those chickens immunized with modified BC/HA in which residues 271–330 from the F protein had been replaced with the corresponding sequence from the NDV AKO strain conferred complete protection against challenge virus shedding. Our findings suggest that this modified rNDV can be used safely as a vaccine vector with enhanced replication, expression, and protective efficacy in avian species, and potentially in humans. PMID:24968158

  19. A molecular tool for detection and tracking of a potential indigenous Beauveria bassiana strain for managing emerald ash borer populations in Canada.

    PubMed

    Johny, Shajahan; Kyei-Poku, George

    2014-10-01

    Emerald ash borer is an invasive species from Asia. Beauveria bassiana strain L49-1AA is being tested for the control of emerald ash borer in Canada, using an autocontamination trapping system. We have developed a simplified allele discrimination polymerase chain reaction (PCR) assay to screen B. bassiana strain, L49-1AA from other Beauveria species by targeting the inter-strain genetic differences in 5' end of EF1-α gene of the genus Beauveria. A single nucleotide polymorphism (SNP) site, T→C was identified only in L49-1AA and was used to develop a simplified allele discrimination polymerase chain reaction (PCR) assay based on a modified allelic inhibition of displacement activity (AIDA) approach for distinguishing B. bassiana L49-1AA from all background Beauveria isolates. The SNP site was employed to design inner primers but with a deliberate mismatch introduced at the 3' antepenultimate from the mutation site in order to maximize specificity and detection efficiency. Amplification was specific to L49-1AA without cross-reaction with DNA from other Beauveria strains. In addition, the designed primers were also tested against environmental samples in L49-1AA released plots and observed to be highly efficient in detecting and discriminating the target strain, L49-1AA from both pure and crude DNA samples. This new method can potentially allow for more discriminatory tracking and monitoring of released L49-1AA in our autocontamination and dissemination projects for managing EAB populations. Additionally, the modified-AIDA format has potential as a tool for simultaneously identifying and differentiating closely related Beauveria species, strains/isolates as well as general classification of other pathogens or organisms. Crown Copyright © 2014. Published by Elsevier Inc. All rights reserved.

  20. An improved Armstrong-Frederick-Type Plasticity Model for Stable Cyclic Stress-Strain Responses Considering Nonproportional Hardening

    NASA Astrophysics Data System (ADS)

    Li, Jing; Zhang, Zhong-ping; Li, Chun-wang

    2018-03-01

    This paper modified an Armstrong-Frederick-type plasticity model for investigating the stable cyclic deformation behavior of metallic materials with different sensitivity to nonproportional loadings. In the modified model, the nonproportionality factor and nonproportional cyclic hardening coefficient coupled with the Jiang-Sehitoglu incremental plasticity model were used to estimate the stable stress-strain responses of the two materials (1045HR steel and 304 stainless steel) under various tension-torsion strain paths. A new equation was proposed to calculate the nonproportionality factor on the basis of the minimum normal strain range. Procedures to determine the minimum normal strain range were presented for general multiaxial loadings. Then, the modified model requires only the cyclic strain hardening exponent and cyclic strength coefficient to determine the material constants. It is convenient for predicting the stable stress-strain responses of materials in engineering application. Comparisons showed that the modified model can reflect the effect of nonproportional cyclic hardening well.

  1. The ability to entrain to long photoperiods differs between 3 Drosophila melanogaster wild-type strains and is modified by twilight simulation.

    PubMed

    Rieger, Dirk; Peschel, Nicolai; Dusik, Verena; Glotz, Silvia; Helfrich-Förster, Charlotte

    2012-02-01

    The ability to adapt to different environmental conditions including seasonal changes is a key feature of the circadian clock. Here, we compared the ability of 3 Drosophila melanogaster wild-type strains to adapt rhythmic activity to long photoperiods simulated in the laboratory. Fruit flies are predominantly crepuscular with activity bouts in the morning (M) and evening (E). The M peak follows dawn and the E peak follows dusk when the photoperiod is extended. We show that this ability is restricted to a certain extension of the phase angle between M and E peaks, such that the E peak does not delay beyond a certain phase under long days. We demonstrate that this ability is significantly improved by simulated twilight and that it depends additionally on the genetic background and the ambient temperature. At 20 °C, the laboratory strain CantonS had the most flexible phase angle between M and E peaks, a Northern wild-type strain had an intermediate one, and a Southern wild-type strain had the lowest flexibility. Furthermore, we found that the 3 strains differed in clock light sensitivity, with the CantonS and the Northern strains more light sensitive than the Southern strain. These results are generally in accord with the recently discovered polymorphisms in the timeless gene (tim) that affect clock light sensitivity.

  2. Microminiature high-resolution linear displacement sensor for peak strain detection in smart structures

    NASA Astrophysics Data System (ADS)

    Arms, Steven W.; Guzik, David C.; Townsend, Christopher P.

    1998-07-01

    Critical civil and military structures require 'smart' sensors in order to report their strain histories; this can help to insure safe operation after exposure to potentially damaging loads. A passive resetable peak strain detector was developed by modifying the mechanics of a differential variable reluctance transducer. The peak strain detector was attached to an aluminum test beam along with a bonded resistance strain gauge and a standard DVRT. Strain measurements were recorded during cyclic beam deflections. DVRT output was compared to the bonded resistance strain gauge output, yielding correlation coefficients ranging from 0.9989 to 0.9998 for al teste, including re-attachment of the DVRT to the specimen. Peak bending strains were obtained by the modified peak detect DVRT to the specimen. Peak bending strains were obtained by the modified peak detect DVRT and this was compared to the peak bending strains as measured by the bonded strain gauge. The peak detect DVRT demonstrated an accuracy of approximately +/- 5 percent over a peak range of 2000 to 2800 microstrain.

  3. BALB/c alleles at modifier loci increase the severity of the maternal effect of the "DDK syndrome".

    PubMed Central

    Le Bras, S; Cohen-Tannoudji, M; Kress, C; Vandormael-Pournin, S; Babinet, C; Baldacci, P

    2000-01-01

    The Om locus was first described in the DDK inbred mouse strain: DDK mice carry a mutation at Om resulting in a parental effect lethality of F(1) embryos. When DDK females are mated with males of other (non-DDK) inbred strains, e.g., BALB/c, they exhibit a low fertility, whereas the reciprocal cross, non-DDK females x DDK males, is fertile (as is the DDK intrastrain cross). The low fertility is due to the death of (DDK x non-DDK)F(1) embryos at the late-morula to blastocyst stage, which is referred to as the "DDK syndrome." The death of these F(1) embryos is caused by an incompatibility between a DDK maternal factor and the non-DDK paternal pronucleus. Previous genetic studies showed that F(1) mice have an intermediate phenotype compared to parental strains: crosses between F(1) females and non-DDK males are semisterile, as are crosses between DDK females and F(1) males. In the present studies, we have examined the properties of mice heterozygous for BALB/c and DDK Om alleles on an essentially BALB/c genetic background. Surprisingly, we found that the females are quasi-sterile when mated with BALB/c males and, thus, present a phenotype similar to DDK females. These results indicate that BALB/c alleles at modifier loci increase the severity of the DDK syndrome. PMID:10655231

  4. A Strategy To Isolate Modifiers of Caenorhabditis elegans Lethal Mutations: Investigating the Endoderm Specifying Ability of the Intestinal Differentiation GATA Factor ELT-2.

    PubMed

    Wiesenfahrt, Tobias; Duanmu, Jingjie; Snider, Frances; Moerman, Don; Au, Vinci; Li-Leger, Erica; Flibotte, Stephane; Parker, Dylan M; Marshall, Craig J; Nishimura, Erin Osborne; Mains, Paul E; McGhee, James D

    2018-05-04

    The ELT-2 GATA factor normally functions in differentiation of the C. elegans endoderm, downstream of endoderm specification. We have previously shown that, if ELT-2 is expressed sufficiently early, it is also able to specify the endoderm and to replace all other members of the core GATA-factor transcriptional cascade (END-1, END-3, ELT-7). However, such rescue requires multiple copies (and presumably overexpression) of the end-1p :: elt-2 cDNA transgene; a single copy of the transgene does not rescue. We have made this observation the basis of a genetic screen to search for genetic modifiers that allow a single copy of the end-1p :: elt-2 cDNA transgene to rescue the lethality of the end-1 end-3 double mutant. We performed this screen on a strain that has a single copy insertion of the transgene in an end-1 end-3 background. These animals are kept alive by virtue of an extrachromosomal array containing multiple copies of the rescuing transgene; the extrachromosomal array also contains a toxin under heat shock control to counterselect for mutagenized survivors that have been able to lose the rescuing array. A screen of ∼14,000 mutagenized haploid genomes produced 17 independent surviving strains. Whole genome sequencing was performed to identify genes that incurred independent mutations in more than one surviving strain. The C. elegans gene tasp-1 was mutated in four independent strains. tasp-1 encodes the C. elegans homolog of Taspase, a threonine-aspartic acid protease that has been found, in both mammals and insects, to cleave several proteins involved in transcription, in particular MLL1/trithorax and TFIIA. A second gene, pqn-82 , was mutated in two independent strains and encodes a glutamine-asparagine rich protein. tasp-1 and pqn-82 were verified as loss-of-function modifiers of the end-1p :: elt-2 transgene by RNAi and by CRISPR/Cas9-induced mutations. In both cases, gene loss leads to modest increases in the level of ELT-2 protein in the early endoderm although ELT-2 levels do not strictly correlate with rescue. We suggest that tasp-1 and pqn-82 represent a class of genes acting in the early embryo to modulate levels of critical transcription factors or to modulate the responsiveness of critical target genes. The screen's design, rescuing lethality with an extrachromosomal transgene followed by counterselection, has a background survival rate of <10 -4 without mutagenesis and should be readily adapted to the general problem of identifying suppressors of C. elegans lethal mutations. Copyright © 2018 Wiesenfahrt et al.

  5. Genome dynamics and evolution in yeasts: A long-term yeast-bacteria competition experiment

    PubMed Central

    Katz, Michael; Knecht, Wolfgang; Compagno, Concetta; Piškur, Jure

    2018-01-01

    There is an enormous genetic diversity evident in modern yeasts, but our understanding of the ecological basis of such diversifications in nature remains at best fragmented so far. Here we report a long-term experiment mimicking a primordial competitive environment, in which yeast and bacteria co-exist and compete against each other. Eighteen yeasts covering a wide phylogenetic background spanning approximately 250 million years of evolutionary history were used to establish independent evolution lines for at most 130 passages. Our collection of hundreds of modified strains generated through such a rare two-species cross-kingdom competition experiment re-created the appearance of large-scale genomic rearrangements and altered phenotypes important in the diversification history of yeasts. At the same time, the methodology employed in this evolutionary study would also be a non-gene-technological method of reprogramming yeast genomes and then selecting yeast strains with desired traits. Cross-kingdom competition may therefore be a method of significant value to generate industrially useful yeast strains with new metabolic traits. PMID:29624585

  6. Rapid genetic typing of diarrheagenic Escherichia coli using a two-tube modified molecular beacon based multiplex real-time PCR assay and its clinical application

    PubMed Central

    2014-01-01

    Background Diarrheagenic Escherichia coli (DEC), including Enterotoxigenic E.coli (ETEC), Enteroaggregative E.coli (EAEC), Enteropathogenic E.coli (EPEC), Enterohemolysin E.coli (EHEC) and Enteroinvasive E.coli (EIEC) causes diarrhea or hemolytic uremic syndromes among infants and travelers around the world. A rapid, reliable and repeatable method is urgent for identifying DEC so as to provide the reference for responding to diarrheal disease outbreak and the treatment of the diarrheal patients associated with DEC. Methods In this study, specific primers and modified molecular beacon probes of nine specific virulence genes, whose 5′end were added with homo tail sequence, were designed; and a two-tube modified molecular beacon based multiplex real–time PCR (rtPCR) assay for the identification of five Escherichia coli pathotypes, including ETEC, EAEC, EPEC, EHEC and EIEC was developed and optimized. Totally 102 bacterial strains, including 52 reference bacterial strains and 50 clinical strains were detected to confirm whether the target genes selected were specific. Then detection limits of the assay were tested. Lastly, the assay was applied to the detection of 11860 clinical samples to evaluate the specificity and sensitivity of the developed assay compared with the conventional PCR. Results The target genes were 100% specific as assessed on 102 bacterial strains since no cross-reactions were observed. The detection limits ranged from 88 CFU/mL (EHEC) to 880 CFU/mL (EPEC). Compared with the conventional PCR, the specificity and sensitivity of the multiplex rtPCR was 100% and over 99%, respectively. The coefficient of variation (CV) for each target gene ranged from 0.45% to 1.53%. 171 positive clinical samples were mostly identified as ETEC (n = 111, 64.9%) and EPEC (n = 38, 22.2%), which were the dominating pathotypes of DEC strains. Conclusion The developed multiplex rtPCR assay for the identification of DEC was high sensitive and specific and could be applied to the rapid identification of DEC in clinical and public health laboratories. PMID:25023669

  7. Genetic engineering of industrial Saccharomyces cerevisiae strains using a selection/counter-selection approach.

    PubMed

    Kutyna, Dariusz R; Cordente, Antonio G; Varela, Cristian

    2014-01-01

    Gene modification of laboratory yeast strains is currently a very straightforward task thanks to the availability of the entire yeast genome sequence and the high frequency with which yeast can incorporate exogenous DNA into its genome. Unfortunately, laboratory strains do not perform well in industrial settings, indicating the need for strategies to modify industrial strains to enable strain development for industrial applications. Here we describe approaches we have used to genetically modify industrial strains used in winemaking.

  8. Investigation of a Coupled Arrhenius-Type/Rossard Equation of AH36 Material.

    PubMed

    Qin, Qin; Tian, Ming-Liang; Zhang, Peng

    2017-04-13

    High-temperature tensile testing of AH36 material in a wide range of temperatures (1173-1573 K) and strain rates (10 -4 -10 -2 s -1 ) has been obtained by using a Gleeble system. These experimental stress-strain data have been adopted to develop the constitutive equation. The constitutive equation of AH36 material was suggested based on the modified Arrhenius-type equation and the modified Rossard equation respectively. The results indicate that the constitutive equation is strongly influenced by temperature and strain, especially strain. Moreover, there is a good agreement between the predicted data of the modified Arrhenius-type equation and the experimental results when the strain is greater than 0.02. There is also good agreement between the predicted data of the Rossard equation and the experimental results when the strain is less than 0.02. Therefore, a coupled equation where the modified Arrhenius-type equation and Rossard equation are combined has been proposed to describe the constitutive equation of AH36 material according to the different strain values in order to improve the accuracy. The correlation coefficient between the computed and experimental flow stress data was 0.998. The minimum value of the average absolute relative error shows the high accuracy of the coupled equation compared with the two modified equations.

  9. Characterization of unknown genetic modifications using high throughput sequencing and computational subtraction

    PubMed Central

    Tengs, Torstein; Zhang, Haibo; Holst-Jensen, Arne; Bohlin, Jon; Butenko, Melinka A; Kristoffersen, Anja Bråthen; Sorteberg, Hilde-Gunn Opsahl; Berdal, Knut G

    2009-01-01

    Background When generating a genetically modified organism (GMO), the primary goal is to give a target organism one or several novel traits by using biotechnology techniques. A GMO will differ from its parental strain in that its pool of transcripts will be altered. Currently, there are no methods that are reliably able to determine if an organism has been genetically altered if the nature of the modification is unknown. Results We show that the concept of computational subtraction can be used to identify transgenic cDNA sequences from genetically modified plants. Our datasets include 454-type sequences from a transgenic line of Arabidopsis thaliana and published EST datasets from commercially relevant species (rice and papaya). Conclusion We believe that computational subtraction represents a powerful new strategy for determining if an organism has been genetically modified as well as to define the nature of the modification. Fewer assumptions have to be made compared to methods currently in use and this is an advantage particularly when working with unknown GMOs. PMID:19814792

  10. Investigation of a Coupled Arrhenius-Type/Rossard Equation of AH36 Material

    PubMed Central

    Qin, Qin; Tian, Ming-Liang; Zhang, Peng

    2017-01-01

    High-temperature tensile testing of AH36 material in a wide range of temperatures (1173–1573 K) and strain rates (10−4–10−2 s−1) has been obtained by using a Gleeble system. These experimental stress-strain data have been adopted to develop the constitutive equation. The constitutive equation of AH36 material was suggested based on the modified Arrhenius-type equation and the modified Rossard equation respectively. The results indicate that the constitutive equation is strongly influenced by temperature and strain, especially strain. Moreover, there is a good agreement between the predicted data of the modified Arrhenius-type equation and the experimental results when the strain is greater than 0.02. There is also good agreement between the predicted data of the Rossard equation and the experimental results when the strain is less than 0.02. Therefore, a coupled equation where the modified Arrhenius-type equation and Rossard equation are combined has been proposed to describe the constitutive equation of AH36 material according to the different strain values in order to improve the accuracy. The correlation coefficient between the computed and experimental flow stress data was 0.998. The minimum value of the average absolute relative error shows the high accuracy of the coupled equation compared with the two modified equations. PMID:28772767

  11. Genetically determined heterogeneity of lung disease in a mouse model of airway mucus obstruction

    PubMed Central

    Grubb, Barbara R.; Kelly, Elizabeth J.; Wilkinson, Kristen J.; Yang, Huifang; Geiser, Marianne; Randell, Scott H.; Boucher, Richard C.; O'Neal, Wanda K.

    2012-01-01

    Mucus clearance is an important airway innate defense mechanism. Airway-targeted overexpression of the epithelial Na+ channel β-subunit [encoded by sodium channel nonvoltage gated 1, beta subunit (Scnn1b)] in mice [Scnn1b-transgenic (Tg) mice] increases transepithelial Na+ absorption and dehydrates the airway surface, which produces key features of human obstructive lung diseases, including mucus obstruction, inflammation, and air-space enlargement. Because the first Scnn1b-Tg mice were generated on a mixed background, the impact of genetic background on disease phenotype in Scnn1b-Tg mice is unknown. To explore this issue, congenic Scnn1b-Tg mice strains were generated on C57BL/6N, C3H/HeN, BALB/cJ, and FVB/NJ backgrounds. All strains exhibited a two- to threefold increase in tracheal epithelial Na+ absorption, and all developed airway mucus obstruction, inflammation, and air-space enlargement. However, there were striking differences in neonatal survival, ranging from 5 to 80% (FVB/NJ

  12. A Modified Constitutive Model for Tensile Flow Behaviors of BR1500HS Ultra-High-Strength Steel at Medium and Low Temperature Regions

    NASA Astrophysics Data System (ADS)

    Zhao, Jun; Quan, Guo-Zheng; Pan, Jia; Wang, Xuan; Wu, Dong-Sen; Xia, Yu-Feng

    2018-01-01

    Constitutive model of materials is one of the most requisite mathematical model in the finite element analysis, which describes the relationships of flow behaviors with strain, strain rate and temperature. In order to construct such constitutive relationships of ultra-high-strength BR1500HS steel at medium and low temperature regions, the true stress-strain data over a wide temperature range of 293-873 K and strain rate range of 0.01-10 s-1 were collected from a series of isothermal uniaxial tensile tests. The experimental results show that stress-strain relationships are highly non-linear and susceptible to three parameters involving temperature, strain and strain rate. By considering the impacts of strain rate and temperature on strain hardening, a modified constitutive model based on Johnson-Cook model was proposed to characterize flow behaviors in medium and low temperature ranges. The predictability of the improved model was also evaluated by the relative error (W(%)), correlation coefficient (R) and average absolute relative error (AARE). The R-value and AARE-value for modified constitutive model at medium and low temperature regions are 0.9915 & 1.56 % and 0.9570 & 5.39 %, respectively, which indicates that the modified constitutive model can precisely estimate the flow behaviors for BR1500HS steel in the medium and low temperature regions.

  13. Origin of the low-frequency internal friction background of gold

    NASA Astrophysics Data System (ADS)

    Baur, J.; Benoit, W.

    1986-11-01

    The internal friction (IF) background of gold is studied in the kHz frequency range. Systematic measurements of IF as a function of frequency, strain amplitude, and temperature show that the IF is due to the superposition of two contributions: the thermoelastic effect and a dislocation effect. The thermoelastic effect is responsible for the IF background observed when the strain amplitude tends to zero. It is the only contribution to the IF background which is strain amplitude independent. On the contrary, the dislocation effect contributes only to the strain amplitude-dependent IF background. This effect is proportional to the strain amplitude. In particular, it is zero when the strain amplitude tends to zero. Furthermore, the dislocation contribution is frequency independent. The experimental results show that the dislocation effect cannot be explained by a viscous damping of dislocation motion, but must be related to an hysteretic and athermal motion of dislocations.

  14. Sex effects in mouse prion disease incubation time.

    PubMed

    Akhtar, Shaheen; Wenborn, Adam; Brandner, Sebastian; Collinge, John; Lloyd, Sarah E

    2011-01-01

    Prion disease incubation time in mice is determined by many factors including PrP expression level, Prnp alleles, genetic background, prion strain and route of inoculation. Sex differences have been described in age of onset for vCJD and in disease duration for both vCJD and sporadic CJD and have also been shown in experimental models. The sex effects reported for mouse incubation times are often contradictory and detail only one strain of mice or prions, resulting in broad generalisations and a confusing picture. To clarify the effect of sex on prion disease incubation time in mice we have compared male and female transmission data from twelve different inbred lines of mice inoculated with at least two prion strains, representing both mouse-adapted scrapie and BSE. Our data show that sex can have a highly significant difference on incubation time. However, this is limited to particular mouse and prion strain combinations. No sex differences were seen in endogenous PrP(C) levels nor in the neuropathological markers of prion disease: PrP(Sc) distribution, spongiosis, neuronal loss and gliosis. These data suggest that when comparing incubation times between experimental groups, such as testing the effects of modifier genes or therapeutics, single sex groups should be used.

  15. Significant competitive advantage conferred by meiosis and syngamy in the yeast Saccharomyces cerevisiae.

    PubMed Central

    Birdsell, J; Wills, C

    1996-01-01

    The presumed advantages of genetic recombinations are difficult to demonstrate directly. To investigate the effects of recombination and background heterozygosity on competitive ability, we have performed serial-transfer competition experiments between isogenic sexual and asexual strains of the yeast Saccharomyces cerevisiae. The members of these diploid pairs of strains differed only in being heterozygous (sexual) or homozygous (asexual) at the mating type or MAT locus. Competing pairs had either a completely homozygous or a heterozygous genetic background, the latter being heterozygous at many different loci throughout the genome. A round of meiotic recombination (automixis) conferred a large and statistically significant enhancement of competitive ability on sexual strains with a heterozygous genetic background. By contrast, in homozygous background competitions, meiosis decreased the sexual strains' initial relative competitive ability. In all cases, however, the sexual strains outcompeted their isogenic asexual counterparts, whether meiotic recombination had occurred or not. In some genetic backgrounds, this was due in part to an overdominance effect on competitive advantage of heterozygosity at the MAT locus. The advantage of the sexual strains also increased significantly during the course of the homozygous background competitions, particularly when meiosis had occurred. This latter effect either did not occur or was very weak in heterozygous background competitions. Overall, sexual strains with heterozygous genetic backgrounds had a significantly higher initial relative competitive ability than those with homozygous backgrounds. The advantage of mating type heterozygosity in this organism extends far beyond the ability to recombine meiotically. PMID:8570658

  16. Systematic Validation and Atomic Force Microscopy of Non-Covalent Short Oligonucleotide Barcode Microarrays

    PubMed Central

    Cook, Michael A.; Chan, Chi-Kin; Jorgensen, Paul; Ketela, Troy; So, Daniel; Tyers, Mike; Ho, Chi-Yip

    2008-01-01

    Background Molecular barcode arrays provide a powerful means to analyze cellular phenotypes in parallel through detection of short (20–60 base) unique sequence tags, or “barcodes”, associated with each strain or clone in a collection. However, costs of current methods for microarray construction, whether by in situ oligonucleotide synthesis or ex situ coupling of modified oligonucleotides to the slide surface are often prohibitive to large-scale analyses. Methodology/Principal Findings Here we demonstrate that unmodified 20mer oligonucleotide probes printed on conventional surfaces show comparable hybridization signals to covalently linked 5′-amino-modified probes. As a test case, we undertook systematic cell size analysis of the budding yeast Saccharomyces cerevisiae genome-wide deletion collection by size separation of the deletion pool followed by determination of strain abundance in size fractions by barcode arrays. We demonstrate that the properties of a 13K unique feature spotted 20 mer oligonucleotide barcode microarray compare favorably with an analogous covalently-linked oligonucleotide array. Further, cell size profiles obtained with the size selection/barcode array approach recapitulate previous cell size measurements of individual deletion strains. Finally, through atomic force microscopy (AFM), we characterize the mechanism of hybridization to unmodified barcode probes on the slide surface. Conclusions/Significance These studies push the lower limit of probe size in genome-scale unmodified oligonucleotide microarray construction and demonstrate a versatile, cost-effective and reliable method for molecular barcode analysis. PMID:18253494

  17. Identification of prostate cancer modifier pathways using parental strain expression mapping

    PubMed Central

    Xu, Qing; Majumder, Pradip K.; Ross, Kenneth; Shim, Yeonju; Golub, Todd R.; Loda, Massimo; Sellers, William R.

    2007-01-01

    Inherited genetic risk factors play an important role in cancer. However, other than the Mendelian fashion cancer susceptibility genes found in familial cancer syndromes, little is known about risk modifiers that control individual susceptibility. Here we developed a strategy, parental strain expression mapping, that utilizes the homogeneity of inbred mice and genome-wide mRNA expression analyses to directly identify candidate germ-line modifier genes and pathways underlying phenotypic differences among murine strains exposed to transgenic activation of AKT1. We identified multiple candidate modifier pathways and, specifically, the glycolysis pathway as a candidate negative modulator of AKT1-induced proliferation. In keeping with the findings in the murine models, in multiple human prostate expression data set, we found that enrichment of glycolysis pathways in normal tissues was associated with decreased rates of cancer recurrence after prostatectomy. Together, these data suggest that parental strain expression mapping can directly identify germ-line modifier pathways of relevance to human disease. PMID:17978178

  18. Phenotypic detection of broad-spectrum beta-lactamases in microbiological practice.

    PubMed

    Htoutou Sedlakova, Miroslava; Hanulik, Vojtech; Chroma, Magdalena; Hricova, Kristyna; Kolar, Milan; Latal, Tomas; Schaumann, Reiner; Rodloff, Arne C

    2011-05-01

    Enterobacteriaceae producing ESBL and AmpC enzymes can be associated with failure of antibiotic therapy and related morbidity and mortality. Their routine detection in microbiology laboratories is still a problem. The aim of this study was to compare the sensitivity of selected phenotypic methods. A total of 106 strains of the Enterobacteriaceae family were tested, in which molecular biology methods confirmed the presence of genes encoding ESBL or AmpC. In ESBL-positive strains, the sensitivity of the ESBL Etest (AB Biodisk) and a modified double-disk synergy test (DDST) were evaluated. AmpC strains were tested by a modified AmpC disk method using 3-aminophenylboronic acid. For simultaneous detection of ESBL and AmpC, the microdilution method with a modified set of antimicrobial agents was used. The sensitivity of the ESBL Etest was 95%; the modified DDST yielded 100% sensitivity for ESBL producers and the AmpC test correctly detected 95% of AmpC-positive strains. The sensitivity of the modified microdilution method was 87% and 95% for ESBL and AmpC beta lactamases, respectively. The detection of ESBL and AmpC beta lactamases should be based on specific phenotypic methods such as the modified DDST, ESBL Etest, AmpC disk test and the modified microdilution method.

  19. Consomic mouse strain selection based on effect size measurement, statistical significance testing and integrated behavioral z-scoring: focus on anxiety-related behavior and locomotion.

    PubMed

    Labots, M; Laarakker, M C; Ohl, F; van Lith, H A

    2016-06-29

    Selecting chromosome substitution strains (CSSs, also called consomic strains/lines) used in the search for quantitative trait loci (QTLs) consistently requires the identification of the respective phenotypic trait of interest and is simply based on a significant difference between a consomic and host strain. However, statistical significance as represented by P values does not necessarily predicate practical importance. We therefore propose a method that pays attention to both the statistical significance and the actual size of the observed effect. The present paper extends on this approach and describes in more detail the use of effect size measures (Cohen's d, partial eta squared - η p (2) ) together with the P value as statistical selection parameters for the chromosomal assignment of QTLs influencing anxiety-related behavior and locomotion in laboratory mice. The effect size measures were based on integrated behavioral z-scoring and were calculated in three experiments: (A) a complete consomic male mouse panel with A/J as the donor strain and C57BL/6J as the host strain. This panel, including host and donor strains, was analyzed in the modified Hole Board (mHB). The consomic line with chromosome 19 from A/J (CSS-19A) was selected since it showed increased anxiety-related behavior, but similar locomotion compared to its host. (B) Following experiment A, female CSS-19A mice were compared with their C57BL/6J counterparts; however no significant differences and effect sizes close to zero were found. (C) A different consomic mouse strain (CSS-19PWD), with chromosome 19 from PWD/PhJ transferred on the genetic background of C57BL/6J, was compared with its host strain. Here, in contrast with CSS-19A, there was a decreased overall anxiety in CSS-19PWD compared to C57BL/6J males, but not locomotion. This new method shows an improved way to identify CSSs for QTL analysis for anxiety-related behavior using a combination of statistical significance testing and effect sizes. In addition, an intercross between CSS-19A and CSS-19PWD may be of interest for future studies on the genetic background of anxiety-related behavior.

  20. Evaluation of a genetically modified foot-and-mouth disease virus vaccine candidate generated by reverse genetics

    PubMed Central

    2012-01-01

    Background Foot-and-mouth disease (FMD) is the most economically important and highly contagious disease of cloven-hoofed animals worldwide. Control of the disease has been mainly based on large-scale vaccinations with whole-virus inactivated vaccines. In recent years, a series of outbreaks of type O FMD occurred in China (including Chinese Taipei, Chinese Hong Kong) posed a tremendous threat to Chinese animal husbandry. Its causative agent, type O FMDV, has evolved into three topotypes (East–South Asia (ME-SA), Southeast Asia (SEA), Cathay (CHY)) in these regions, which represents an important obstacle to disease control. The available FMD vaccine in China shows generally good protection against ME-SA and SEA topotype viruses infection, but affords insufficient protection against some variants of the CHY topotype. Therefore, the choice of a new vaccine strain is of fundamental importance. Results The present study describes the generation of a full-length infectious cDNA clone of FMDV vaccine strain and a genetically modified virus with some amino acid substitutions in antigenic sites 1, 3, and 4, based on the established infectious clone. The recombinant viruses had similar growth properties to the wild O/HN/CHA/93 virus. All swine immunized with inactivated vaccine prepared from the O/HN/CHA/93 were fully protected from challenge with the viruses of ME-SA and SEA topotypes and partially protected against challenge with the virus of CHY topotype at 28 days post-immunization. In contrast, the swine inoculated with the genetically modified vaccine were completely protected from the infection of viruses of the three topotypes. Conclusions Some amino acid substitutions in the FMDV vaccine strain genome did not have an effect on the ability of viral replication in vitro. The vaccine prepared from genetically modified FMDV by reverse genetics significantly improved the protective efficacy to the variant of the CHY topotype, compared with the wild O/HN/CHA/93 virus. Thus, the full-length cDNA clone of FMDV can be a useful tool to develop genetically engineered FMDV vaccine candidates to help control porcinophilic FMD epidemics in China. PMID:22591597

  1. The amyR-deletion strain of Aspergillus niger CICC2462 is a suitable host strain to express secreted protein with a low background.

    PubMed

    Zhang, Hui; Wang, Shuang; Zhang, Xiang Xiang; Ji, Wei; Song, Fuping; Zhao, Yue; Li, Jie

    2016-04-28

    The filamentous fungus Aspergillus niger is widely exploited as an important expression host for industrial production. The glucoamylase high-producing strain A. niger CICC2462 has been used as a host strain for the establishment of a secretion expression system. It expresses recombinant xylanase, mannase and asparaginase at a high level, but some high secretory background proteins in these recombinant strains still remain, such as alpha-amylase and alpha-glucosidase; lead to a low-purity of fermentation products. The aim was to construct an A. niger host strain with a low background of protein secretion. The transcription factor amyR was deleted in A. niger CICC2462, and the results from enzyme activity assays and SDS-PAGE analysis showed that the glucoamylase and amylase activities of the ∆amyR strains were significantly lower than those of the wild-type strain. High-throughput RNA-sequencing and shotgun LC-MS/MS proteomic technology analysis demonstrated that the expression of amylolytic enzymes was decreased at both the transcriptional and translational levels in the ∆amyR strain. Interestingly, the ∆amyR strain growth rate better than the wild-type strain. Our findings clearly indicated that the ∆amyR strain of A. niger CICC2462 can be used as a host strain with a low background of protein secretion.

  2. Laboratory evolution of copper tolerant yeast strains

    PubMed Central

    2012-01-01

    Background Yeast strains endowed with robustness towards copper and/or enriched in intracellular Cu might find application in biotechnology processes, among others in the production of functional foods. Moreover, they can contribute to the study of human diseases related to impairments of copper metabolism. In this study, we investigated the molecular and physiological factors that confer copper tolerance to strains of baker's yeasts. Results We characterized the effects elicited in natural strains of Candida humilis and Saccharomyces cerevisiae by the exposure to copper in the culture broth. We observed that, whereas the growth of Saccharomyces cells was inhibited already at low Cu concentration, C. humilis was naturally robust and tolerated up to 1 g · L-1 CuSO4 in the medium. This resistant strain accumulated over 7 mg of Cu per gram of biomass and escaped severe oxidative stress thanks to high constitutive levels of superoxide dismutase and catalase. Both yeasts were then "evolved" to obtain hyper-resistant cells able to proliferate in high copper medium. While in S. cerevisiae the evolution of robustness towards Cu was paralleled by the increase of antioxidative enzymes, these same activities decreased in evolved hyper-resistant Candida cells. We also characterized in some detail changes in the profile of copper binding proteins, that appeared to be modified by evolution but, again, in a different way in the two yeasts. Conclusions Following evolution, both Candida and Saccharomyces cells were able to proliferate up to 2.5 g · L-1 CuSO4 and to accumulate high amounts of intracellular copper. The comparison of yeasts differing in their robustness, allowed highlighting physiological and molecular determinants of natural and acquired copper tolerance. We observed that different mechanisms contribute to confer metal tolerance: the control of copper uptake, changes in the levels of enzymes involved in oxidative stress response and changes in the copper-binding proteome. However, copper elicits different physiological and molecular reactions in yeasts with different backgrounds. PMID:22214286

  3. A high-fat diet differentially regulates glutathione phenotypes in the obesity-prone mouse strains DBA/2J, C57BL/6J, and AKR/J.

    PubMed

    Norris, Katie M; Okie, Whitney; Kim, Woo Kyun; Adhikari, Roshan; Yoo, Sarah; King, Stephanie; Pazdro, Robert

    2016-12-01

    The ubiquitous tripeptide glutathione (GSH) is a critical component of the endogenous antioxidant defense system. Tissue GSH concentrations and redox status (GSH/GSSG) are genetically controlled, but it is unclear whether interactions between genetic background and diet affect GSH homeostasis. The current study tested the hypothesis that a high-fat diet regulates GSH homeostasis in a manner dependent on genetic background. At 4 months of age, female mice representing 3 obesity-prone inbred strains-C57BL/6J (B6), DBA/2J (D2), and AKR/J (AKR)-were randomly assigned to consume a control (10% energy from fat) or high-fat (62% energy from fat) diet for 10 weeks (n=5/diet per strain). Tissue GSH levels, GSSG levels, and GSH/GSSG were quantified, and hepatic expression of GSH-related enzymes was evaluated by quantitative reverse transcription polymerase chain reaction. The high-fat diet caused a decrease in hepatic GSH/GSSG in D2 mice. In contrast, B6 mice exhibited a decrease in GSSG levels in the liver and kidney, as well as a resultant increase in renal GSH/GSSG. AKR mice also exhibited increased renal GSH/GSSG on a high-fat diet. Finally, the high-fat diet induced a unique gene expression response in D2 mice compared with B6 and AKR. The D2 response was characterized by up-regulation of glutamate-cysteine ligase modifier subunit and down-regulation of glutathione reductase, whereas the B6 and AKR responses were characterized by up-regulation of glutathione peroxidase 1. Two-way analysis of variance analyses confirmed several diet-strain interactions within the GSH system, and linear regression models highlighted relationships between body mass and GSH outcomes as well. Overall, our data indicate that dietary fat regulates the GSH system in a strain-dependent manner. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Relationship between Job Stress and Hypo-high-density Lipoproteinemia of Chinese Workers in Shanghai: The Rosai Karoshi Study

    PubMed Central

    Muratsubaki, Tomohiko; Hattori, Tomomi; Li, Jue; Fukudo, Shin; Munakata, Masanori

    2016-01-01

    Background: Karoshi, or death due to overwork, has now become a serious social problem in China. Worsening of cardiovascular risks by stress might initiate karoshi. Many studies have examined the relationship between job stress and obesity, hypertension, and type 2 diabetes mellitus, but less evidence exists for dyslipidemia like hypo-high-density lipoproteinemia (hypo-HDL). The aim of this study was to investigate the relationship between job stress and hypo-HDL of Chinese workers in Shanghai. Methods: We studied 2219 Chinese workers in Shanghai, who participated in the Japan-China cooperative study for the prevention of karoshi. A questionnaire was administered to examine the lifestyle characteristics, job category, weekly working hours, and job stress. Job demand and job control were quantified using the National Institute for Occupational Safety and Health questionnaire. Modified job strain measure was defined by the combination of low job control and high demand. Hypo-HDL was defined as plasma high-density lipoprotein cholesterol concentration of <1.04 mmol/L (40 mg/dl). Multivariate logistic regression analysis was performed for hypo-HDL as a dependent variable. Results: Modified job strain was not related to hypo-HDL either in men or women. In men, multivariate adjusted odds ratio (OR) for having hypo-HDL was significantly higher in the lowest job control tertile compared with the highest job control tertile (OR = 1.39, 95% confidence interval [CI] 1.03–1.87, P = 0.034). In the same model, a similar trend was observed for women, but it did not reach a statistically significant level (OR = 1.51, 95% CI, 0.88–2.56, P = 0.132). Conclusion: A low level of job control but not modified job strain was significantly related to higher prevalence of hypo-HDL of Chinese workers in Shanghai. PMID:27748331

  5. A modified live canine parvovirus strain with novel plaque characteristics. I. Viral attenuation and dog response.

    PubMed

    Carmichael, L E; Joubert, J C; Pollock, R V

    1981-10-01

    A canine parvovirus (CPV) strain (C-780916) was found attenuated for pups at 80, but not after 51 serial passages in dog kidney cell (DKC) cultures. A variant viral population ('large plaque') emerged after prolonged cultivation in DKC cultures that may be associated with reduced native virulence. Dogs vaccinated with modified CPV developed high hemagglutination-inhibiting (HI) antibody titers within 4 days of incoluation and antibody persisted. Vaccinated animals shed small amounts of virus in the feces that spread to contact dogs. After five back-passages in dogs the modified strain was not pathogenic for pups and the plaque characteristics of the virus isolated from the feces were typical of the attenuated strain. The modified live CPV did not cause infection of the fetus when inoculated parenterally into pregnant bitches at various stages of gestation. It was not pathogenic for neonatal pups. These results suggest that a safe and effective live homologous (CPV) vaccine has been developed which should aid substantially in controlling CPV infection.

  6. Cloning and expression of vgb gene in Bacillus cereus, improve phenol and p-nitrophenol biodegradation

    NASA Astrophysics Data System (ADS)

    Vélez-Lee, Angel Eduardo; Cordova-Lozano, Felipe; Bandala, Erick R.; Sanchez-Salas, Jose Luis

    2016-02-01

    In this work, the vgb gene from Vitrocilla stercoraria was used to genetically modify a Bacillus cereus strain isolated from pulp and paper wastewater effluent. The gene was cloned in a multicopy plasmid (pUB110) or uni-copy gene using a chromosome integrative vector (pTrpBG1). B. cereus and its recombinant strains were used for phenol and p-nitrophenol biodegradation using aerobic or micro-aerobic conditions and two different temperatures (i.e. 37 and 25 °C). Complete (100%) phenol degradation was obtained for the strain where the multicopy of vgb gene was present, 98% for the strain where uni-copy gene was present and 45% for wild type strain for the same experimental conditions (i.e. 37 °C and aerobic condition). For p-nitrophenol degradation at the same conditions, the strain with the multi-copy vgb gene was capable to achieve 50% of biodegradation, ˜100% biodegradation was obtained using the uni-copy strain and ˜24% for wild type strain. When the micro-aerobic condition was tested, the biodegradation yield showed a significant decreased. The biodegradation trend observed for aerobic was similar for micro-aerobic assessments: the modified strains showed higher degradation rates when compared with wild type strain. For all experimental conditions, the highest p-nitrophenol degradation was observed using the strain with uni-copy of vgb gene. Besides the increase of biodegradative capability of the strain, insertion of the vgb gene was observed able to modify other morphological characteristics such as avoiding the typical flake formation in the B. cereus culture. In both cases, the modification seems to be related with the enhancement of oxygen supply to the cells generated by the vgb gene insertion. The application of the genetically modified microorganism (GMM) to the biodegradation of pollutants in contaminated water possesses high potential as an environmentally friendly technology to facing this emergent problem.

  7. Improving membrane protein expression and function using genomic edits

    DOE PAGES

    Jensen, Heather M.; Eng, Thomas; Chubukov, Victor; ...

    2017-10-12

    Expression of membrane proteins often leads to growth inhibition and perturbs central metabolism and this burden varies with the protein being overexpressed. There are also known strain backgrounds that allow greater expression of membrane proteins but that differ in efficacy across proteins. Here, we hypothesized that for any membrane protein, it may be possible to identify a modified strain background where its expression can be accommodated with less burden. To directly test this hypothesis, we used a bar-coded transposon insertion library in tandem with cell sorting to assess genome-wide impact of gene deletions on membrane protein expression. The expression ofmore » five membrane proteins (CyoB, CydB, MdlB, YidC, and LepI) and one soluble protein (GST), each fused to GFP, was examined. We identified Escherichia coli mutants that demonstrated increased membrane protein expression relative to that in wild type. For two of the proteins (CyoB and CydB), we conducted functional assays to confirm that the increase in protein expression also led to phenotypic improvement in function. This study represents a systematic approach to broadly identify genetic loci that can be used to improve membrane protein expression, and our method can be used to improve expression of any protein that poses a cellular burden.« less

  8. Improving membrane protein expression and function using genomic edits

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

    Jensen, Heather M.; Eng, Thomas; Chubukov, Victor

    Expression of membrane proteins often leads to growth inhibition and perturbs central metabolism and this burden varies with the protein being overexpressed. There are also known strain backgrounds that allow greater expression of membrane proteins but that differ in efficacy across proteins. Here, we hypothesized that for any membrane protein, it may be possible to identify a modified strain background where its expression can be accommodated with less burden. To directly test this hypothesis, we used a bar-coded transposon insertion library in tandem with cell sorting to assess genome-wide impact of gene deletions on membrane protein expression. The expression ofmore » five membrane proteins (CyoB, CydB, MdlB, YidC, and LepI) and one soluble protein (GST), each fused to GFP, was examined. We identified Escherichia coli mutants that demonstrated increased membrane protein expression relative to that in wild type. For two of the proteins (CyoB and CydB), we conducted functional assays to confirm that the increase in protein expression also led to phenotypic improvement in function. This study represents a systematic approach to broadly identify genetic loci that can be used to improve membrane protein expression, and our method can be used to improve expression of any protein that poses a cellular burden.« less

  9. A Modified Mechanical Threshold Stress Constitutive Model for Austenitic Stainless Steels

    NASA Astrophysics Data System (ADS)

    Prasad, K. Sajun; Gupta, Amit Kumar; Singh, Yashjeet; Singh, Swadesh Kumar

    2016-12-01

    This paper presents a modified mechanical threshold stress (m-MTS) constitutive model. The m-MTS model incorporates variable athermal and dynamic strain aging (DSA) Components to accurately predict the flow stress behavior of austenitic stainless steels (ASS)-316 and 304. Under strain rate variations between 0.01-0.0001 s-1, uniaxial tensile tests were conducted at temperatures ranging from 50-650 °C to evaluate the material constants of constitutive models. The test results revealed the high dependence of flow stress on strain, strain rate and temperature. In addition, it was observed that DSA occurred at elevated temperatures and very low strain rates, causing an increase in flow stress. While the original MTS model is capable of predicting the flow stress behavior for ASS, statistical parameters point out the inefficiency of the model when compared to other models such as Johnson Cook model, modified Zerilli-Armstrong (m-ZA) model, and modified Arrhenius-type equations (m-Arr). Therefore, in order to accurately model both the DSA and non-DSA regimes, the original MTS model was modified by incorporating variable athermal and DSA components. The suitability of the m-MTS model was assessed by comparing the statistical parameters. It was observed that the m-MTS model was highly accurate for the DSA regime when compared to the existing models. However, models like m-ZA and m-Arr showed better results for the non-DSA regime.

  10. Replacing Escherichia coli NAD-dependent glyceraldehyde 3-phosphate dehydrogenase (GAPDH) with a NADP-dependent enzyme from Clostridium acetobutylicum facilitates NADPH dependent pathways.

    PubMed

    Martínez, Irene; Zhu, Jiangfeng; Lin, Henry; Bennett, George N; San, Ka-Yiu

    2008-11-01

    Reactions requiring reducing equivalents, NAD(P)H, are of enormous importance for the synthesis of industrially valuable compounds such as carotenoids, polymers, antibiotics and chiral alcohols among others. The use of whole-cell biocatalysis can reduce process cost by acting as catalyst and cofactor regenerator at the same time; however, product yields might be limited by cofactor availability within the cell. Thus, our study focussed on the genetic manipulation of a whole-cell system by modifying metabolic pathways and enzymes to improve the overall production process. In the present work, we genetically engineered an Escherichia coli strain to increase NADPH availability to improve the productivity of products that require NADPH in its biosynthesis. The approach involved an alteration of the glycolysis step where glyceraldehyde-3-phosphate (GAP) is oxidized to 1,3 bisphophoglycerate (1,3-BPG). This reaction is catalyzed by NAD-dependent endogenous glyceraldehyde-3-phosphate dehydrogenase (GAPDH) encoded by the gapA gene. We constructed a recombinant E. coli strain by replacing the native NAD-dependent gapA gene with a NADP-dependent GAPDH from Clostridium acetobutylicum, encoded by the gene gapC. The beauty of this approach is that the recombinant E. coli strain produces 2 mol of NADPH, instead of NADH, per mole of glucose consumed. Metabolic flux analysis showed that the flux through the pentose phosphate (PP) pathway, one of the main pathways that produce NADPH, was reduced significantly in the recombinant strain when compared to that of the parent strain. The effectiveness of the NADPH enhancing system was tested using the production of lycopene and epsilon-caprolactone as model systems using two different background strains. The recombinant strains, with increased NADPH availability, consistently showed significant higher productivity than the parent strains.

  11. Wild Sex in Zebrafish: Loss of the Natural Sex Determinant in Domesticated Strains

    PubMed Central

    Wilson, Catherine A.; High, Samantha K.; McCluskey, Braedan M.; Amores, Angel; Yan, Yi-lin; Titus, Tom A.; Anderson, Jennifer L.; Batzel, Peter; Carvan, Michael J.; Schartl, Manfred; Postlethwait, John H.

    2014-01-01

    Sex determination can be robustly genetic, strongly environmental, or genetic subject to environmental perturbation. The genetic basis of sex determination is unknown for zebrafish (Danio rerio), a model for development and human health. We used RAD-tag population genomics to identify sex-linked polymorphisms. After verifying this “RAD-sex” method on medaka (Oryzias latipes), we studied two domesticated zebrafish strains (AB and TU), two natural laboratory strains (WIK and EKW), and two recent isolates from nature (NA and CB). All four natural strains had a single sex-linked region at the right tip of chromosome 4, enabling sex genotyping by PCR. Genotypes for the single nucleotide polymorphism (SNP) with the strongest statistical association to sex suggested that wild zebrafish have WZ/ZZ sex chromosomes. In natural strains, “male genotypes” became males and some “female genotypes” also became males, suggesting that the environment or genetic background can cause female-to-male sex reversal. Surprisingly, TU and AB lacked detectable sex-linked loci. Phylogenomics rooted on D. nigrofasciatus verified that all strains are monophyletic. Because AB and TU branched as a monophyletic clade, we could not rule out shared loss of the wild sex locus in a common ancestor despite their independent domestication. Mitochondrial DNA sequences showed that investigated strains represent only one of the three identified zebrafish haplogroups. Results suggest that zebrafish in nature possess a WZ/ZZ sex-determination mechanism with a major determinant lying near the right telomere of chromosome 4 that was modified during domestication. Strains providing the zebrafish reference genome lack key components of the natural sex-determination system but may have evolved variant sex-determining mechanisms during two decades in laboratory culture. PMID:25233988

  12. Bacterial adherence to anodized titanium alloy

    NASA Astrophysics Data System (ADS)

    Pérez-Jorge Peremarch, C.; Pérez Tanoira, R.; Arenas, M. A.; Matykina, E.; Conde, A.; De Damborenea, J. J.; Gómez Barrena, E.; Esteban, J.

    2010-11-01

    The aim of this study was to evaluate Staphylococcus sp adhesion to modified surfaces of anodized titanium alloy (Ti-6Al-4V). Surface modification involved generation of fluoride-containing titanium oxide nanotube films. Specimens of Ti-6Al-4V alloy 6-4 ELI-grade 23- meets the requirements of ASTM F136 2002A (AMS 2631B class A1) were anodized in a mixture of sulphuric/hydrofluoric acid at 20 V for 5 and 60 min to form a 100 nm-thick porous film of 20 nm pore diameter and 230 nm-thick nanotube films of 100 nm in diameter. The amount of fluorine in the oxide films was of 6% and of 4%, respectively. Collection strains and six clinical strains each of Staphylococcus aureus and Staphylococcus epidermidis were studied. The adherence study was performed using a previously published protocol by Kinnari et al. The experiments were performed in triplicates. As a result, lower adherence was detected for collection strains in modified materials than in unmodified controls. Differences between clinical strains were detected for both species (p<0.0001, Kruskal-Wallis test), although global data showed similar results to that of collection strains (p<0.0001, Kruskal-Wallis test). Adherence of bacteria to modified surfaces was decreased for both species. The results also reflect a difference in the adherence between S. aureus and S. epidermidis to the modified material. As a conclusion, not only we were able to confirm the decrease of adherence in the modified surface, but also the need to test multiple clinical strains to obtain more realistic microbiological results due to intraspecies differences.

  13. Implementation of an Associative Flow Rule Including Hydrostatic Stress Effects Into the High Strain Rate Deformation Analysis of Polymer Matrix Composites

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.; Roberts, Gary D.; Gilat, Amos

    2003-01-01

    A previously developed analytical formulation has been modified in order to more accurately account for the effects of hydrostatic stresses on the nonlinear, strain rate dependent deformation of polymer matrix composites. State variable constitutive equations originally developed for metals have been modified in order to model the nonlinear, strain rate dependent deformation of polymeric materials. To account for the effects of hydrostatic stresses, which are significant in polymers, the classical J2 plasticity theory definitions of effective stress and effective inelastic strain, along with the equations used to compute the components of the inelastic strain rate tensor, are appropriately modified. To verify the revised formulation, the shear and tensile deformation of two representative polymers are computed across a wide range of strain rates. Results computed using the developed constitutive equations correlate well with experimental data. The polymer constitutive equations are implemented within a strength of materials based micromechanics method to predict the nonlinear, strain rate dependent deformation of polymer matrix composites. The composite mechanics are verified by analyzing the deformation of a representative polymer matrix composite for several fiber orientation angles across a variety of strain rates. The computed values compare well to experimentally obtained results.

  14. A Modified Double Multiple Nonlinear Regression Constitutive Equation for Modeling and Prediction of High Temperature Flow Behavior of BFe10-1-2 Alloy

    NASA Astrophysics Data System (ADS)

    Cai, Jun; Wang, Kuaishe; Shi, Jiamin; Wang, Wen; Liu, Yingying

    2018-01-01

    Constitutive analysis for hot working of BFe10-1-2 alloy was carried out by using experimental stress-strain data from isothermal hot compression tests, in a wide range of temperature of 1,023 1,273 K, and strain rate range of 0.001 10 s-1. A constitutive equation based on modified double multiple nonlinear regression was proposed considering the independent effects of strain, strain rate, temperature and their interrelation. The predicted flow stress data calculated from the developed equation was compared with the experimental data. Correlation coefficient (R), average absolute relative error (AARE) and relative errors were introduced to verify the validity of the developed constitutive equation. Subsequently, a comparative study was made on the capability of strain-compensated Arrhenius-type constitutive model. The results showed that the developed constitutive equation based on modified double multiple nonlinear regression could predict flow stress of BFe10-1-2 alloy with good correlation and generalization.

  15. The use of genetically modified Saccharomyces cerevisiae strains in the wine industry.

    PubMed

    Schuller, Dorit; Casal, Margarida

    2005-08-01

    In recent decades, science and food technology have contributed at an accelerated rate to the introduction of new products to satisfy nutritional, socio-economic and quality requirements. With the emergence of modern molecular genetics, the industrial importance of Saccharomyces cerevisiae, is continuously extended. The demand for suitable genetically modified (GM) S. cerevisiae strains for the biofuel, bakery and beverage industries or for the production of biotechnological products (e.g. enzymes, pharmaceutical products) will continuously grow in the future. Numerous specialised S. cerevisiae wine strains were obtained in recent years, possessing a wide range of optimised or novel oenological properties, capable of satisfying the demanding nature of modern winemaking practise. The unlocking of transcriptome, proteome and metabolome complexities will contribute decisively to the knowledge about the genetic make-up of commercial yeast strains and will influence wine strain improvement via genetic engineering. The most relevant advances regarding the importance and implications of the use of GM yeast strains in the wine industry are discussed in this mini-review. In this work, various aspects are considered including the strategies used for the construction of strains with respect to current legislation requirements, the environmental risk evaluations concerning the deliberate release of genetically modified yeast strains, the methods for detection of recombinant DNA and protein that are currently under evaluation, and the reasons behind the critical public perception towards the application of such strains.

  16. Manufacturing and shear response characterization of carbon nanofiber modified CFRP using the out-of-autoclave-vacuum-bag-only cure process.

    PubMed

    McDonald, Erin E; Wallace, Landon F; Hickman, Gregory J S; Hsiao, Kuang-Ting

    2014-01-01

    The interlaminar shear response is studied for carbon nanofiber (CNF) modified out-of-autoclave-vacuum-bag-only (OOA-VBO) carbon fiber reinforced plastic (CFRP). Commercial OOA-VBO prepregs were coated with a CNF modified epoxy solution and a control epoxy solution without CNF to make CNF modified samples and control samples, respectively. Tensile testing was used to study the in-plane shear performance of [± 45°]4s composite laminates. Significant difference in failure modes between the control and CNF modified CFRPs was identified. The control samples experienced half-plane interlaminar delamination, whereas the CNF modified samples experienced a localized failure in the intralaminar region. Digital image correlation (DIC) surface strain results of the control sample showed no further surface strain increase along the delaminated section when the sample was further elongated prior to sample failure. On the other hand, the DIC results of the CNF modified sample showed that the surface strain increased relatively and uniformly across the CFRP as the sample was further elongated until sample failure. The failure mode evidence along with microscope pictures indicated that the CNF modification acted as a beneficial reinforcement inhibiting interlaminar delamination.

  17. Manufacturing and Shear Response Characterization of Carbon Nanofiber Modified CFRP Using the Out-of-Autoclave-Vacuum-Bag-Only Cure Process

    PubMed Central

    McDonald, Erin E.; Wallace, Landon F.; Hickman, Gregory J. S.; Hsiao, Kuang-Ting

    2014-01-01

    The interlaminar shear response is studied for carbon nanofiber (CNF) modified out-of-autoclave-vacuum-bag-only (OOA-VBO) carbon fiber reinforced plastic (CFRP). Commercial OOA-VBO prepregs were coated with a CNF modified epoxy solution and a control epoxy solution without CNF to make CNF modified samples and control samples, respectively. Tensile testingwas used to study the in-plane shear performance of [±45°]4s composite laminates. Significant difference in failure modes between the control and CNF modified CFRPs was identified. The control samples experienced half-plane interlaminar delamination, whereas the CNF modified samples experienced a localized failure in the intralaminar region. Digital image correlation (DIC) surface strain results of the control sample showed no further surface strain increase along the delaminated section when the sample was further elongated prior to sample failure. On the other hand, the DIC results of the CNF modified sample showed that the surface strain increased relatively and uniformly across the CFRP as the sample was further elongated until sample failure. The failure mode evidence along with microscope pictures indicated that the CNF modification acted as a beneficial reinforcement inhibiting interlaminar delamination. PMID:24688435

  18. Rescue of the mouse DDK syndrome by parent-of-origin-dependent modifiers.

    PubMed

    Ideraabdullah, Folami Y; Kim, Kuikwon; Pomp, Daniel; Moran, Jennifer L; Beier, David; de Villena, Fernando Pardo-Manuel

    2007-02-01

    When females of the DDK inbred mouse strain are mated to males of other strains, 90-100% of the resulting embryos die during early embryonic development. This DDK syndrome lethality results from incompatibility between an ooplasmic DDK factor and a non-DDK paternal gene, which map to closely linked loci on chromosome 11. It has been proposed that the expression of the gene that encodes the ooplasmic factor is subject to allelic exclusion in oocytes. Previous studies have demonstrated the existence of recessive modifiers that increase lethality in the C57BL/6 and BALB/c strains. These modifiers are thought to skew the choice of allele undergoing allelic exclusion in the oocytes of heterozygous females. In the present study, we demonstrate the presence of modifiers in three Mus musculus domesticus wild-derived strains, PERA, PERC, and RBA. These modifiers completely rescued DDK syndrome lethality. We mapped the major locus that is responsible for rescue in PERA and PERC crosses to proximal chromosome 13 and named this locus Rmod1 (Rescue Modifier of the DDK Syndrome 1). Our experiments demonstrate that PERA or PERC alleles at Rmod1 rescue lethality independently of allelic exclusion. In addition, rescue of the lethal phenotype depends on the parental origin of the Rmod1 alleles; transmission through the dam leads to rescue, while transmission through the sire has no effect.

  19. High throughput screening of CO2-tolerating microalgae using GasPak bags

    PubMed Central

    2013-01-01

    Background Microalgae are diverse in terms of their speciation and function. More than 35,000 algal strains have been described, and thousands of algal cultures are maintained in different culture collection centers. The ability of CO2 uptake by microalgae varies dramatically among algal species. It becomes challenging to select suitable algal candidates that can proliferate under high CO2 concentration from a large collection of algal cultures. Results Here, we described a high throughput screening method to rapidly identify high CO2 affinity microalgae. The system integrates a CO2 mixer, GasPak bags and microplates. Microalgae on the microplates will be cultivated in GasPak bags charged with different CO2 concentrations. Using this method, we identified 17 algal strains whose growth rates were not influenced when the concentration of CO2 was increased from 2 to 20% (v/v). Most CO2 tolerant strains identified in this study were closely related to the species Scenedesmus and Chlorococcum. One of Scenedesmus strains (E7A) has been successfully tested in in the scale up photo bioreactors (500 L) bubbled with flue gas which contains 10-12% CO2. Conclusion Our high throughput CO2 testing system provides a rapid and reliable way for identifying microalgal candidate strains that can grow under high CO2 condition from a large pool of culture collection species. This high throughput system can also be modified for selecting algal strains that can tolerate other gases, such as NOx, SOx, or flue gas. PMID:24341988

  20. Strain-Gradient Modulated Exciton Emission in Bent ZnO Wires Probed by Cathodoluminescence.

    PubMed

    Fu, Xue-Wen; Li, Cai-Zhen; Fang, Liang; Liu, Da-Meng; Xu, Jun; Yu, Da-Peng; Liao, Zhi-Min

    2016-12-27

    Photoelectrical properties of semiconductor nanostructures are expected to be improved significantly by strain engineering. Besides the local strain, the strain gradient is promising to tune the luminescence properties by modifying the crystal symmetry. Here, we report the investigation of strain-gradient induced symmetry-breaking effect on excitonic states in pure bending ZnO microwires by high spatial-resolved cathodoluminescence at low temperature of 80 K. In addition to the local-strain induced light emission peak shift, the bound exciton emission photon energy shows an extraordinary jump of ∼16.6 meV at a high strain-gradient of 1.22% μm -1 , which is ascribed to the strain gradient induced symmetry-breaking. Such a symmetry-breaking lifts the energy degeneracy of the electronic band structures, which significantly modifies the electron-hole interactions and the fine structures of the bound exciton states. These results provide a further understanding of the strain gradient effect on the excitonic states and possess a potential for the applications in optoelectronic devices.

  1. Anti-Biofilm Activity of Polyazolidinammonium Modified with Iodine Hydrate Ions against Microbial Biofilms of Uropathogenic Coliform Bacteria.

    PubMed

    Nechaeva, O V; Tikhomirova, E I; Zayarsky, D A; Bespalova, N V; Glinskaya, E V; Shurshalova, N F; Al Bayati, B M; Babailova, A I

    2017-04-01

    The dynamics of microbial biofilm formation by standard strain and by clinical strains of uropathogenic coliform bacteria was investigated in vitro and the effect of sublethal concentrations of the polymer compound polyazolidinammonium modified with iodine hydrate ions on the initial stages of biofilm formation was assessed. Treatment of immunological plate wells with the polymeric compound prevented film formation, especially in case of clinical E. coli strain carrying FimH virulence gene.

  2. Toughening mechanism in elastometer-modified epoxy resins: Part 1

    NASA Technical Reports Server (NTRS)

    Yee, A. F.; Pearson, R. A.

    1983-01-01

    Several plaques of Epon 828, cured with piperidine, modified with hycar(r) CTBN 1300X8, Hycar(R) CTBN 1300X13, and Hycar(R) CTBN 1300x15, and in some cases modified with biphenol A (BPA), yielded properly toughened epoxies with rubber particle diameters ranging from 0.1 to 10 microns. Fracture toughness experiments indicate that toughness was more a function of rubber content than the rubber particle size. Tensile volumetric behavior of the near resin exhibits two regions: an initial region where the increase in volume strain was due to the Poisson's effect, and a second region where a slower rate of increase in volume strain was due to shear deformation. Tensile volumetric deformation of an elastomer-modified epoxy exhibits the same type of behavior to that of the neat resin at low rates ( 3.2x0.01 sec(-1)). But at very high strain rates, which correspond more closely to the strain rates at the crack tip, there exists an increase in volume strain beyond the Poisson's effect. TEM, SEM and OM studies indicate that the rubber particles had voided. When a thin section from the deformed region is viewed under crossed-polarized light, shear bands are seen connecting voided rubber particles. From this information cavitation and enhanced shear band formation is proposed as the toughening mechanism.

  3. Comparative proteomic profiles of Aspergillus fumigatus and Aspergillus lentulus strains by surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS)

    PubMed Central

    2011-01-01

    Background Surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) was applied to analyze the protein profiles in both somatic and metabolic extracts of Aspergillus species. The study was carried out on some Aspergillus species within the Fumigati section (Aspergillus fumigatus wild-types and natural abnormally pigmented mutants, and Aspergillus lentulus). The aim was to validate whether mass spectrometry protein profiles can be used as specific signatures to discriminate different Aspergillus species or even mutants within the same species. Results The growth conditions and the SELDI-TOF parameters were determined to generate characteristic protein profiles of somatic and metabolic extracts of Aspergillus fumigatus strains using five different ProteinChips®, eight growth conditions combining two temperatures, two media and two oxygenation conditions. Nine strains were investigated: three wild-types and four natural abnormally pigmented mutant strains of A. fumigatus and two strains of A. lentulus. A total of 242 fungal extracts were prepared. The spectra obtained are protein signatures linked to the physiological states of fungal strains depending on culture conditions. The best resolutions were obtained using the chromatographic surfaces CM10, NP20 and H50 with fractions of fungi grown on modified Sabouraud medium at 37°C in static condition. Under these conditions, the SELDI-TOF analysis allowed A. fumigatus and A. lentulus strains to be grouped into distinct clusters. Conclusions SELDI-TOF analysis distinguishes A. fumigatus from A. lentulus strains and moreover, permits separate clusters of natural abnormally pigmented A. fumigatus strains to be obtained. In addition, this methodology allowed us to point out fungal components specifically produced by a wild-type strain or natural mutants. It offers attractive potential for further studies of the Aspergillus biology or pathogenesis. PMID:21798007

  4. Strain Engineering to Modify the Electrochemistry of Energy Storage Electrodes

    PubMed Central

    Muralidharan, Nitin; Carter, Rachel; Oakes, Landon; Cohn, Adam P.; Pint, Cary L.

    2016-01-01

    Strain engineering has been a critical aspect of device design in semiconductor manufacturing for the past decade, but remains relatively unexplored for other applications, such as energy storage. Using mechanical strain as an input parameter to modulate electrochemical potentials of metal oxides opens new opportunities intersecting fields of electrochemistry and mechanics. Here we demonstrate that less than 0.1% strain on a Ni-Ti-O based metal-oxide formed on superelastic shape memory NiTi alloys leads to anodic and cathodic peak potential shifts by up to ~30 mV in an electrochemical cell. Moreover, using the superelastic properties of NiTi to enable strain recovery also recovers the electrochemical potential of the metal oxide, providing mechanistic evidence of strain-modified electrochemistry. These results indicate that mechanical energy can be coupled with electrochemical systems to efficiently design and optimize a new class of strain-modulated energy storage materials. PMID:27283872

  5. Establishment and comparison of four constitutive relationships of PC/ABS from low to high uniaxial strain rates

    NASA Astrophysics Data System (ADS)

    Wang, Haitao; Zhang, Yun; Huang, Zhigao; Tang, Zhongbin; Wang, Yanpei; Zhou, Huamin

    2017-10-01

    The objective of this paper is to accurately predict the rate/temperature-dependent deformation of a polycarbonate (PC) and acrylonitrile-butadiene-styrene (ABS) blend at low, moderate, and high strain rates for various temperatures. Four constitutive models have been employed to predict stress-strain responses of PC/ABS under these conditions, including the DSGZ model, the original Mulliken-Boyce (M-B) model, the modified M-B model, and an adiabatic model named the Wang model. To more accurately capture the large deformation of PC/ABS under the high strain rate loading, the original M-B model is modified by allowing for the evolution of the internal shear strength. All of the four constitutive models above have been implemented in the finite element software ABAQUS/Explicit. A comparison of prediction accuracies of the four constitutive models over a wide range of strain rates and temperatures has been presented. The modified M-B model is observed to be more accurate in predicting the deformation of PC/ABS at high strain rates for various temperatures than the original M-B model, and the Wang model is demonstrated to be the most accurate in simulating the deformation of PC/ABS at low, moderate, and high strain rates for various temperatures.

  6. Altering the selection capabilities of common cloning vectors via restriction enzyme mediated gene disruption

    PubMed Central

    2013-01-01

    Background The cloning of gene sequences forms the basis for many molecular biological studies. One important step in the cloning process is the isolation of bacterial transformants carrying vector DNA. This involves a vector-encoded selectable marker gene, which in most cases, confers resistance to an antibiotic. However, there are a number of circumstances in which a different selectable marker is required or may be preferable. Such situations can include restrictions to host strain choice, two phase cloning experiments and mutagenesis experiments, issues that result in additional unnecessary cloning steps, in which the DNA needs to be subcloned into a vector with a suitable selectable marker. Results We have used restriction enzyme mediated gene disruption to modify the selectable marker gene of a given vector by cloning a different selectable marker gene into the original marker present in that vector. Cloning a new selectable marker into a pre-existing marker was found to change the selection phenotype conferred by that vector, which we were able to demonstrate using multiple commonly used vectors and multiple resistance markers. This methodology was also successfully applied not only to cloning vectors, but also to expression vectors while keeping the expression characteristics of the vector unaltered. Conclusions Changing the selectable marker of a given vector has a number of advantages and applications. This rapid and efficient method could be used for co-expression of recombinant proteins, optimisation of two phase cloning procedures, as well as multiple genetic manipulations within the same host strain without the need to remove a pre-existing selectable marker in a previously genetically modified strain. PMID:23497512

  7. Elevated temperature axial and torsional fatigue behavior of Haynes 188

    NASA Technical Reports Server (NTRS)

    Bonacuse, Peter J.; Kalluri, Sreeramesh

    1995-01-01

    The results are reported for high-temperature axial and torsional low-cycle fatigue experiments performed at 760 C in air on thin-walled tubular specimens of Haynes 188, a wrought cobalt-based superalloy. Data are also presented for mean coefficient of thermal expansion, elastic modulus, and shear modulus at various temperatures from room to 1000 C, and monotonic and cyclic stress-strain curves in tension and in shear at 760 C. This data set is used to evaluate several multiaxial fatigue life models (most were originally developed for room temperature multiaxial life prediction) including von Mises equivalent strain range (ASME Boiler and Pressure Code), Manson-Halford, modified multiaxiality factor (proposed in this paper), modified Smith-Watson-Topper, and Fatemi-Socie-Kurath. At von Mises equivalent strain ranges (the torsional strain range divided by the square root of 3, taking the Poisson's ratio to be 0.5), torsionally strained specimens lasted, on average, factors of 2 to 3 times longer than axially strained specimens. The modified multiaxiality factor approach shows promise as a useful method of estimating torsional fatigue life from axial fatigue data at high temperatures. Several difficulties arose with the specimen geometry and extensometry used in these experiments. Cracking at extensometer probe indentations was a problem at smaller strain ranges. Also, as the largest axial and torsional strain range fatigue tests neared completion, a small amount of specimen buckling was observed.

  8. Non-Mendelian Female Sterility in DROSOPHILA MELANOGASTER: Influence of Aging and Thermic Treatments. III. Cumulative Effects Induced by These Factors

    PubMed Central

    Bucheton, Alain

    1979-01-01

    Crosses between various strains of Drosophila melanogaster may give rise to a female sterility of non-Mendelian determination. Reduced fertility is observed in females, known as SF females, bred from crosses between females of "reactive" strains and males of "inducer" strains. The reduced fertility of the SF females is the result of an interaction between an extrachromosomal property, the reactivity, and a chromosomal factor, I. The extrachromosomal property varies considerably in its ability to reduce fertility. The fertility reduction of the SF females corresponds to what is known as the reactivity level of their reactive mothers. Two nongenetic factors can modify the level of reactivity: aging and temperature. The action of aging is cumulative. When the flies of a reactive strain are submitted at each generation to the action of this factor, the level of reactivity of this strain is gradually modified. The modifications induced are reversible. Indeed, when such a modified strain is returned to standard breeding conditions, the reactivity returns progressively to its initial level. The effect of thermic treatments also seems to be cumulative and reversible. PMID:121289

  9. Elevated temperature axial and torsional fatigue behavior of Haynes 188

    NASA Astrophysics Data System (ADS)

    Bonacuse, Peter J.; Kalluri, Sreeramesh

    1992-06-01

    The results of high-temperature axial and torsional low-cycle fatigue experiments performed on Haynes 188, a wrought cobalt-base superalloy, are reported. Fatigue tests were performed at 760 C in air on thin-walled tubular specimens at various ranges under strain control. Data are also presented for coefficient of thermal expansion, elastic modulus, and shear modulus at various temperatures from room to 1000 C, and monotonic and cyclic stress-strain curves in tension and in shear at 760 C. The data set is used to evaluate several multiaxial fatigue life models (most were originally developed for room temperature multiaxial life prediction) including von Mises equivalent strain range (ASME boiler and pressure vessel code), Manson-Halford, Modified Multiaxiality Factor (proposed here), Modified Smith-Watson-Topper, and Fatemi-Socie-Kurath. At von Mises equivalent strain ranges (the torsional strain range divided by the square root of 3, taking the Poisson's ratio to be 0.5), torsionally strained specimens lasted, on average, factors of 2 to 3 times longer than axially strained specimens. The Modified Multiaxiality Factor approach shows promise as a useful method of estimating torsional fatigue life from axial fatigue data at high temperatures. Several difficulties arose with the specimen geometry and extensometry used in these experiments. Cracking at extensometer probe indentations was a problem at smaller strain ranges. Also, as the largest axial and torsional strain range fatigue tests neared completion, a small amount of specimen buckling was observed.

  10. Constitutive Modeling of the High-Temperature Flow Behavior of α-Ti Alloy Tube

    NASA Astrophysics Data System (ADS)

    Lin, Yanli; Zhang, Kun; He, Zhubin; Fan, Xiaobo; Yan, Yongda; Yuan, Shijian

    2018-04-01

    In the hot metal gas forming process, the deformation conditions, such as temperature, strain rate and deformation degree, are often prominently changed. The understanding of the flow behavior of α-Ti seamless tubes over a relatively wide range of temperatures and strain rates is important. In this study, the stress-strain curves in the temperature range of 973-1123 K and the initial strain rate range of 0.0004-0.4 s-1 were measured by isothermal tensile tests to conduct a constitutive analysis and a deformation behavior analysis. The results show that the flow stress decreases with the decrease in the strain rate and the increase of the deformation temperature. The Fields-Backofen model and Fields-Backofen-Zhang model were used to describe the stress-strain curves. The Fields-Backofen-Zhang model shows better predictability on the flow stress than the Fields-Backofen model, but there exists a large deviation in the deformation condition of 0.4 s-1. A modified Fields-Backofen-Zhang model is proposed, in which a strain rate term is introduced. This modified Fields-Backofen-Zhang model gives a more accurate description of the flow stress variation under hot forming conditions with a higher strain rate up to 0.4 s-1. Accordingly, it is reasonable to adopt the modified Fields-Backofen-Zhang model for the hot forming process which is likely to reach a higher strain rate, such as 0.4 s-1.

  11. Elevated temperature axial and torsional fatigue behavior of Haynes 188

    NASA Technical Reports Server (NTRS)

    Bonacuse, Peter J.; Kalluri, Sreeramesh

    1992-01-01

    The results of high-temperature axial and torsional low-cycle fatigue experiments performed on Haynes 188, a wrought cobalt-base superalloy, are reported. Fatigue tests were performed at 760 C in air on thin-walled tubular specimens at various ranges under strain control. Data are also presented for coefficient of thermal expansion, elastic modulus, and shear modulus at various temperatures from room to 1000 C, and monotonic and cyclic stress-strain curves in tension and in shear at 760 C. The data set is used to evaluate several multiaxial fatigue life models (most were originally developed for room temperature multiaxial life prediction) including von Mises equivalent strain range (ASME boiler and pressure vessel code), Manson-Halford, Modified Multiaxiality Factor (proposed here), Modified Smith-Watson-Topper, and Fatemi-Socie-Kurath. At von Mises equivalent strain ranges (the torsional strain range divided by the square root of 3, taking the Poisson's ratio to be 0.5), torsionally strained specimens lasted, on average, factors of 2 to 3 times longer than axially strained specimens. The Modified Multiaxiality Factor approach shows promise as a useful method of estimating torsional fatigue life from axial fatigue data at high temperatures. Several difficulties arose with the specimen geometry and extensometry used in these experiments. Cracking at extensometer probe indentations was a problem at smaller strain ranges. Also, as the largest axial and torsional strain range fatigue tests neared completion, a small amount of specimen buckling was observed.

  12. Constitutive Modeling of the High-Temperature Flow Behavior of α-Ti Alloy Tube

    NASA Astrophysics Data System (ADS)

    Lin, Yanli; Zhang, Kun; He, Zhubin; Fan, Xiaobo; Yan, Yongda; Yuan, Shijian

    2018-05-01

    In the hot metal gas forming process, the deformation conditions, such as temperature, strain rate and deformation degree, are often prominently changed. The understanding of the flow behavior of α-Ti seamless tubes over a relatively wide range of temperatures and strain rates is important. In this study, the stress-strain curves in the temperature range of 973-1123 K and the initial strain rate range of 0.0004-0.4 s-1 were measured by isothermal tensile tests to conduct a constitutive analysis and a deformation behavior analysis. The results show that the flow stress decreases with the decrease in the strain rate and the increase of the deformation temperature. The Fields-Backofen model and Fields-Backofen-Zhang model were used to describe the stress-strain curves. The Fields-Backofen-Zhang model shows better predictability on the flow stress than the Fields-Backofen model, but there exists a large deviation in the deformation condition of 0.4 s-1. A modified Fields-Backofen-Zhang model is proposed, in which a strain rate term is introduced. This modified Fields-Backofen-Zhang model gives a more accurate description of the flow stress variation under hot forming conditions with a higher strain rate up to 0.4 s-1. Accordingly, it is reasonable to adopt the modified Fields-Backofen-Zhang model for the hot forming process which is likely to reach a higher strain rate, such as 0.4 s-1.

  13. Characterization of the Metabolically Modified Heavy Metal-Resistant Cupriavidus metallidurans Strain MSR33 Generated for Mercury Bioremediation

    PubMed Central

    Rojas, Luis A.; Yáñez, Carolina; González, Myriam; Lobos, Soledad; Smalla, Kornelia; Seeger, Michael

    2011-01-01

    Background Mercury-polluted environments are often contaminated with other heavy metals. Therefore, bacteria with resistance to several heavy metals may be useful for bioremediation. Cupriavidus metallidurans CH34 is a model heavy metal-resistant bacterium, but possesses a low resistance to mercury compounds. Methodology/Principal Findings To improve inorganic and organic mercury resistance of strain CH34, the IncP-1β plasmid pTP6 that provides novel merB, merG genes and additional other mer genes was introduced into the bacterium by biparental mating. The transconjugant Cupriavidus metallidurans strain MSR33 was genetically and biochemically characterized. Strain MSR33 maintained stably the plasmid pTP6 over 70 generations under non-selective conditions. The organomercurial lyase protein MerB and the mercuric reductase MerA of strain MSR33 were synthesized in presence of Hg2+. The minimum inhibitory concentrations (mM) for strain MSR33 were: Hg2+, 0.12 and CH3Hg+, 0.08. The addition of Hg2+ (0.04 mM) at exponential phase had not an effect on the growth rate of strain MSR33. In contrast, after Hg2+ addition at exponential phase the parental strain CH34 showed an immediate cessation of cell growth. During exposure to Hg2+ no effects in the morphology of MSR33 cells were observed, whereas CH34 cells exposed to Hg2+ showed a fuzzy outer membrane. Bioremediation with strain MSR33 of two mercury-contaminated aqueous solutions was evaluated. Hg2+ (0.10 and 0.15 mM) was completely volatilized by strain MSR33 from the polluted waters in presence of thioglycolate (5 mM) after 2 h. Conclusions/Significance A broad-spectrum mercury-resistant strain MSR33 was generated by incorporation of plasmid pTP6 that was directly isolated from the environment into C. metallidurans CH34. Strain MSR33 is capable to remove mercury from polluted waters. This is the first study to use an IncP-1β plasmid directly isolated from the environment, to generate a novel and stable bacterial strain useful for mercury bioremediation. PMID:21423734

  14. Effect of micro-structural modifier on the morphology of silicon rich secondary phase and strain hardening behavior of eutectic Al-Si alloy

    NASA Astrophysics Data System (ADS)

    Mansoor, M.; Salam, I.; Tauqir, A.

    2016-08-01

    Eutectic Al-Si alloys find their applications in moderate to severe tribological conditions, for example: pistons, casings of high speed pumps and slide sleeves. The higher hardness, so the better tribological properties, are originated by the formation of a silicon rich secondary phase, however, the morphology of the secondary phase drastically influence the toughness of the alloy. Microstructural modifiers are used to control the toughness which modifies the Si rich secondary phase into dispersed spherical structure instead of needle-like network. In the present study, a mixture of chemical fluxes was used to modify the Si phase. The alloy was cast into a sand mold and characterized by scanning electron microscopy, energy dispersive spectroscopy, hardness testing and tensile testing. It was found that the morphology of the Si phase was altered to acicular structure due to the modification process. In comparison, the un-modified alloy contained Si phase in needle-like structure. The effect of modifier was also pronounced on the mechanical properties, where increase of 50% in yield strength, 56% in tensile strength and 200% in elongation occurred. A discernable raise in strain hardening component indicated the improved strain harden ability and formability of the modified alloy.

  15. Leveraging Genetic-Background Effects in Saccharomyces cerevisiae To Improve Lignocellulosic Hydrolysate Tolerance

    DOE PAGES

    Sardi, Maria; Rovinskiy, Nikolay; Zhang, Yaoping; ...

    2016-07-22

    We report a major obstacle to sustainable lignocellulosic biofuel production is microbe inhibition by the combinatorial stresses in pretreated plant hydrolysate. Chemical biomass pretreatment releases a suite of toxins that interact with other stressors, including high osmolarity and temperature, which together can have poorly understood synergistic effects on cells. Improving tolerance in industrial strains has been hindered, in part because the mechanisms of tolerance reported in the literature often fail to recapitulate in other strain backgrounds. Here, we explored and then exploited variations in stress tolerance, toxin-induced transcriptomic responses, and fitness effects of gene overexpression in different Saccharomyces cerevisiae (yeast)more » strains to identify genes and processes linked to tolerance of hydrolysate stressors. Using six different S. cerevisiae strains that together maximized phenotypic and genetic diversity, first we explored transcriptomic differences between resistant and sensitive strains to identify common and strain-specific responses. This comparative analysis implicated primary cellular targets of hydrolysate toxins, secondary effects of defective defense strategies, and mechanisms of tolerance. Dissecting the responses to individual hydrolysate components across strains pointed to synergistic interactions between osmolarity, pH, hydrolysate toxins, and nutrient composition. By characterizing the effects of high-copy gene overexpression in three different strains, we revealed the breadth of the background-specific effects of gene fitness contributions in synthetic hydrolysate. Lastly, our approach identified new genes for engineering improved stress tolerance in diverse strains while illuminating the effects of genetic background on molecular mechanisms.« less

  16. Isolation of Shiga toxin-producing Escherichia coli serogroups O26, O45, O103, O111, O121, and O145 from ground beef using modified rainbow agar and post-immunomagnetic separation acid treatment.

    PubMed

    Tillman, Glenn E; Wasilenko, Jamie L; Simmons, Mustafa; Lauze, Todd A; Minicozzi, Joseph; Oakley, Brian B; Narang, Neelam; Fratamico, Pina; Cray, Ailliam C

    2012-09-01

    It is estimated that at least 70% of human illnesses due to non-O157 Shiga toxin-producing Escherichia coli (STEC) in the United States are caused by strains from the top six serogroups (O26, O45, O103, O111, O121, and O145). Procedures for isolating STEC from food products often use plating media that include antimicrobial supplements at concentrations that inhibit background microflora growth but can also inhibit target STEC growth. In this study, an agar medium with lower supplement concentrations, modified Rainbow agar (mRBA), was evaluated for recovery of STEC serogroups O26, O45, O103, O111, O121, and O145 from ground beef enrichments. A post-immunomagnetic separation (IMS) acid treatment step was additionally used to reduce background microflora and increase recovery of target STEC strains. Ground beef samples (325 g) were artificially contaminated with STEC and confounding organisms and enriched for 15 h. Recovery of the target STEC was attempted on the enrichments using IMS and plating onto mRBA and Rainbow agar (RBA). Additionally, acid treatment was performed on the post-IMS eluate followed by plating onto mRBA. Using the combination of mRBA and acid treatment, target STEC were isolated from 103 (85.8%) of 120 of the low-inoculated samples (1 to 5 CFU/325-g sample) compared with 68 (56.7%) of 120 using no acid treatment and plating onto RBA with higher levels of novobiocin and potassium tellurite. The combination of acid treatment and mRBA provides a significant improvement over the use of RBA for isolation of STEC serogroups O26, O45, O103, O111, O121, and O145 from raw ground beef.

  17. Modification of tRNALys UUU by Elongator Is Essential for Efficient Translation of Stress mRNAs

    PubMed Central

    Sansó, Miriam; Buhne, Karin; Carmona, Mercè; Paulo, Esther; Hermand, Damien; Rodríguez-Gabriel, Miguel; Ayté, José; Leidel, Sebastian; Hidalgo, Elena

    2013-01-01

    The Elongator complex, including the histone acetyl transferase Sin3/Elp3, was isolated as an RNA polymerase II-interacting complex, and cells deficient in Elongator subunits display transcriptional defects. However, it has also been shown that Elongator mediates the modification of some tRNAs, modulating translation efficiency. We show here that the fission yeast Sin3/Elp3 is important for oxidative stress survival. The stress transcriptional program, governed by the Sty1-Atf1-Pcr1 pathway, is affected in mutant cells, but not severely. On the contrary, cells lacking Sin3/Elp3 cannot modify the uridine wobble nucleoside of certain tRNAs, and other tRNA modifying activities such as Ctu1-Ctu2 are also essential for normal tolerance to H2O2. In particular, a plasmid over-expressing the tRNALys UUU complements the stress-related phenotypes of Sin3/Elp3 mutant cells. We have determined that the main H2O2-dependent genes, including those coding for the transcription factors Atf1 and Pcr1, are highly expressed mRNAs containing a biased number of lysine-coding codons AAA versus AAG. Thus, their mRNAs are poorly translated after stress in cells lacking Sin3/Elp3 or Ctu2, whereas a mutated atf1 transcript with AAA-to-AAG lysine codons is efficiently translated in all strain backgrounds. Our study demonstrates that the lack of a functional Elongator complex results in stress phenotypes due to its contribution to tRNA modification and subsequent translation inefficiency of certain stress-induced, highly expressed mRNAs. These results suggest that the transcriptional defects of these strain backgrounds may be a secondary consequence of the deficient expression of a transcription factor, Atf1-Pcr1, and other components of the transcriptional machinery. PMID:23874237

  18. Modification of tRNA(Lys) UUU by elongator is essential for efficient translation of stress mRNAs.

    PubMed

    Fernández-Vázquez, Jorge; Vargas-Pérez, Itzel; Sansó, Miriam; Buhne, Karin; Carmona, Mercè; Paulo, Esther; Hermand, Damien; Rodríguez-Gabriel, Miguel; Ayté, José; Leidel, Sebastian; Hidalgo, Elena

    2013-01-01

    The Elongator complex, including the histone acetyl transferase Sin3/Elp3, was isolated as an RNA polymerase II-interacting complex, and cells deficient in Elongator subunits display transcriptional defects. However, it has also been shown that Elongator mediates the modification of some tRNAs, modulating translation efficiency. We show here that the fission yeast Sin3/Elp3 is important for oxidative stress survival. The stress transcriptional program, governed by the Sty1-Atf1-Pcr1 pathway, is affected in mutant cells, but not severely. On the contrary, cells lacking Sin3/Elp3 cannot modify the uridine wobble nucleoside of certain tRNAs, and other tRNA modifying activities such as Ctu1-Ctu2 are also essential for normal tolerance to H2O2. In particular, a plasmid over-expressing the tRNA(Lys) UUU complements the stress-related phenotypes of Sin3/Elp3 mutant cells. We have determined that the main H2O2-dependent genes, including those coding for the transcription factors Atf1 and Pcr1, are highly expressed mRNAs containing a biased number of lysine-coding codons AAA versus AAG. Thus, their mRNAs are poorly translated after stress in cells lacking Sin3/Elp3 or Ctu2, whereas a mutated atf1 transcript with AAA-to-AAG lysine codons is efficiently translated in all strain backgrounds. Our study demonstrates that the lack of a functional Elongator complex results in stress phenotypes due to its contribution to tRNA modification and subsequent translation inefficiency of certain stress-induced, highly expressed mRNAs. These results suggest that the transcriptional defects of these strain backgrounds may be a secondary consequence of the deficient expression of a transcription factor, Atf1-Pcr1, and other components of the transcriptional machinery.

  19. The strain-rate sensitivity of high-strength high-toughness steels.

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

    Dilmore, M.F.; Crenshaw, Thomas B.; Boyce, Brad Lee

    2006-01-01

    The present study examines the strain-rate sensitivity of four high strength, high-toughness alloys at strain rates ranging from 0.0002 s-1 to 200 s-1: Aermet 100, a modified 4340, modified HP9-4-20, and a recently developed Eglin AFB steel alloy, ES-1c. A refined dynamic servohydraulic method was used to perform tensile tests over this entire range. Each of these alloys exhibit only modest strain-rate sensitivity. Specifically, the strain-rate sensitivity exponent m, is found to be in the range of 0.004-0.007 depending on the alloy. This corresponds to a {approx}10% increase in the yield strength over the 7-orders of magnitude change in strain-rate.more » Interestingly, while three of the alloys showed a concominant {approx}3-10% drop in their ductility with increasing strain-rate, the ES1-c alloy actually exhibited a 25% increase in ductility with increasing strain-rate. Fractography suggests the possibility that at higher strain-rates ES-1c evolves towards a more ductile dimple fracture mode associated with microvoid coalescence.« less

  20. A Comparative Study on Johnson Cook, Modified Zerilli-Armstrong and Arrhenius-Type Constitutive Models to Predict High-Temperature Flow Behavior of Ti-6Al-4V Alloy in α + β Phase

    NASA Astrophysics Data System (ADS)

    Cai, Jun; Wang, Kuaishe; Han, Yingying

    2016-03-01

    True stress and true strain values obtained from isothermal compression tests over a wide temperature range from 1,073 to 1,323 K and a strain rate range from 0.001 to 1 s-1 were employed to establish the constitutive equations based on Johnson Cook, modified Zerilli-Armstrong (ZA) and strain-compensated Arrhenius-type models, respectively, to predict the high-temperature flow behavior of Ti-6Al-4V alloy in α + β phase. Furthermore, a comparative study has been made on the capability of the three models to represent the elevated temperature flow behavior of Ti-6Al-4V alloy. Suitability of the three models was evaluated by comparing both the correlation coefficient R and the average absolute relative error (AARE). The results showed that the Johnson Cook model is inadequate to provide good description of flow behavior of Ti-6Al-4V alloy in α + β phase domain, while the predicted values of modified ZA model and the strain-compensated Arrhenius-type model could agree well with the experimental values except under some deformation conditions. Meanwhile, the modified ZA model could track the deformation behavior more accurately than other model throughout the entire temperature and strain rate range.

  1. Modified Newcastle Disease virus as an improved vaccine vector against Simian Immunodeficiency virus.

    PubMed

    Manoharan, Vinoth K; Khattar, Sunil K; LaBranche, Celia C; Montefiori, David C; Samal, Siba K

    2018-06-12

    SIV infection in macaques is a relevant animal model for HIV pathogenesis and vaccine study in humans. To design a safe and effective vaccine against HIV, we evaluated the suitability of naturally-occurring avirulent Newcastle disease virus (NDV) strains and several modified versions of NDV as vectors for the expression and immunogenicity of SIV envelope protein gp160. All the NDV vectors expressed gp160 protein in infected cells. The gp160 expressed by these vectors formed oligomers and was incorporated into the NDV envelope. All the NDV vectors expressing gp160 were attenuated in chickens. Intranasal immunization of guinea pigs with modified NDV vectors such as rNDV-APMV-2CS/gp160 and rNDV-APMV-8CS/gp160 (NDV strain LaSota containing the cleavage site sequences of F protein of avian paramyxovirus (APMV) serotype 2 and 8, respectively), and rNDV-BC-F-HN/gp160 (NDV strain BC containing LaSota F cleavage site and LaSota F and HN genes) elicited improved SIV-specific humoral and mucosal immune responses compared to other NDV vectors. These modified vectors were also efficient in inducing neutralizing antibody responses to tier 1 A SIVmac251.6 and tier 1B SIVmac251/M766 strains. This study suggests that our novel modified NDV vectors are safe and immunogenic and can be used as vaccine vector to control HIV.

  2. Strain-dependent Differences in LTP and Hippocampus-dependent Memory in Inbred Mice

    PubMed Central

    Nguyen, Peter V.; Abel, Ted; Kandel, Eric R.; Bourtchouladze, Roussoudan

    2000-01-01

    Many studies have used “reverse” genetics to produce “knock-out” and transgenic mice to explore the roles of various molecules in long-term potentiation (LTP) and spatial memory. The existence of a variety of inbred strains of mice provides an additional way of exploring the genetic bases of learning and memory. We examined behavioral memory and LTP expression in area CA1 of hippocampal slices prepared from four different inbred strains of mice: C57BL/6J, CBA/J, DBA/2J, and 129/SvEms-+Ter?/J. We found that LTP induced by four 100-Hz trains of stimulation was robust and long-lasting in C57BL/6J and DBA/2J mice but decayed in CBA/J and 129/SvEms-+Ter?/J mice. LTP induced by one 100-Hz train was significantly smaller after 1 hr in the 129/SvEms-+Ter?/J mice than in the other three strains. Theta-burst LTP was shorter lasting in CBA/J, DBA/2J, and 129/SvEms-+Ter?/J mice than in C57BL/6J mice. We also observed specific memory deficits, among particular mouse strains, in spatial and nonspatial tests of hippocampus-dependent memory. CBA/J mice showed defective learning in the Morris water maze, and both DBA/2J and CBA/J strains displayed deficient long-term memory in contextual and cued fear conditioning tests. Our findings provide strong support for a genetic basis for some forms of synaptic plasticity that are linked to behavioral long-term memory and suggest that genetic background can influence the electrophysiological and behavioral phenotypes observed in genetically modified mice generated for elucidating the molecular bases of learning, memory, and LTP. PMID:10837506

  3. A Congenic Line of the C57BL/6J Mouse Strain that is Proficient in Melatonin Synthesis.

    PubMed

    Zhang, Zhijing; Silveyra, Eduardo; Jin, Nange; Ribelayga, Christophe P

    2018-05-16

    The C57BL/6J (B6) is the most common inbred mouse strain used in biomedical research in the United States. Yet, this strain is notoriously known for being deficient in the biosynthesis of melatonin, an important effector of circadian clocks in the brain and in the retina. Melatonin deficiency in this strain results from non-functional alleles of the genes coding two key enzymes of the melatonin synthesis pathway: arylalkylamine-N-acetyltransferase (Aanat) and N-acetylserotonin-O-methyltransferase (Asmt). By introducing functional alleles of the Aanat and Asmt genes from the melatonin-proficient CBA/CaJ (CBA) mouse strain to B6, we have generated a B6 congenic line that has acquired the capacity of rhythmic melatonin synthesis. In addition, the melatonin-dependent rhythm of dopamine release in the retina is restored in the B6 congenic line. Finally, we have partially characterized the Aanat and Asmt genes of the CBA strain and have identified multiple differences between CBA and B6 alleles, including single nucleotide polymorphism and deletion/insertion of DNA segments of various sizes. As an improved model organism with functional components of the melatonin synthesis pathway and melatonin-dependent circadian regulations, the new line will be useful to researchers studying melatonin physiological functions in a variety of fields including, but not limited to, circadian biology and neuroscience. In particular, the congenic line will be useful to speed up introduction of melatonin production capacity into genetically-modified mouse lines of interest such as knockout lines, many of which are on B6 or mixed B6 backgrounds. The melatonin-proficient B6 congenic line will be widely distributed. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  4. Functional analysis of lipid metabolism genes in wine yeasts during alcoholic fermentation at low temperature

    PubMed Central

    López-Malo, María; García-Ríos, Estéfani; Chiva, Rosana; Guillamon, José M.

    2014-01-01

    Wine produced by low-temperature fermentation is mostly considered to have improved sensory qualities. However few commercial wine strains available on the market are well-adapted to ferment at low temperature (10 - 15°C). The lipid metabolism of Saccharomyces cerevisiae plays a central role in low temperature adaptation. One strategy to modify lipid composition is to alter transcriptional activity by deleting or overexpressing the key genes of lipid metabolism. In a previous study, we identified the genes of the phospholipid, sterol and sphingolipid pathways, which impacted on growth capacity at low temperature. In the present study, we aimed to determine the influence of these genes on fermentation performance and growth during low-temperature wine fermentations. We analyzed the phenotype during fermentation at the low and optimal temperature of the lipid mutant and overexpressing strains in the background of a derivative commercial wine strain. The increase in the gene dosage of some of these lipid genes, e.g., PSD1, LCB3, DPL1 and OLE1, improved fermentation activity during low-temperature fermentations, thus confirming their positive role during wine yeast adaptation to cold. Genes whose overexpression improved fermentation activity at 12°C were overexpressed by chromosomal integration into commercial wine yeast QA23. Fermentations in synthetic and natural grape must were carried out by this new set of overexpressing strains. The strains overexpressing OLE1 and DPL1 were able to finish fermentation before commercial wine yeast QA23. Only the OLE1 gene overexpression produced a specific aroma profile in the wines produced with natural grape must. PMID:28357215

  5. Functional analysis of lipid metabolism genes in wine yeasts during alcoholic fermentation at low temperature.

    PubMed

    López-Malo, María; García-Ríos, Estéfani; Chiva, Rosana; Guillamon, José M

    2014-10-29

    Wine produced by low-temperature fermentation is mostly considered to have improved sensory qualities. However few commercial wine strains available on the market are well-adapted to ferment at low temperature (10 - 15°C). The lipid metabolism of Saccharomyces cerevisiae plays a central role in low temperature adaptation. One strategy to modify lipid composition is to alter transcriptional activity by deleting or overexpressing the key genes of lipid metabolism. In a previous study, we identified the genes of the phospholipid, sterol and sphingolipid pathways, which impacted on growth capacity at low temperature. In the present study, we aimed to determine the influence of these genes on fermentation performance and growth during low-temperature wine fermentations. We analyzed the phenotype during fermentation at the low and optimal temperature of the lipid mutant and overexpressing strains in the background of a derivative commercial wine strain. The increase in the gene dosage of some of these lipid genes, e.g., PSD1 , LCB3, DPL1 and OLE1, improved fermentation activity during low-temperature fermentations, thus confirming their positive role during wine yeast adaptation to cold. Genes whose overexpression improved fermentation activity at 12°C were overexpressed by chromosomal integration into commercial wine yeast QA23. Fermentations in synthetic and natural grape must were carried out by this new set of overexpressing strains. The strains overexpressing OLE1 and DPL1 were able to finish fermentation before commercial wine yeast QA23. Only the OLE1 gene overexpression produced a specific aroma profile in the wines produced with natural grape must.

  6. Xylose Fermentation by Saccharomyces cerevisiae: Challenges and Prospects.

    PubMed

    Moysés, Danuza Nogueira; Reis, Viviane Castelo Branco; de Almeida, João Ricardo Moreira; de Moraes, Lidia Maria Pepe; Torres, Fernando Araripe Gonçalves

    2016-02-25

    Many years have passed since the first genetically modified Saccharomyces cerevisiae strains capable of fermenting xylose were obtained with the promise of an environmentally sustainable solution for the conversion of the abundant lignocellulosic biomass to ethanol. Several challenges emerged from these first experiences, most of them related to solving redox imbalances, discovering new pathways for xylose utilization, modulation of the expression of genes of the non-oxidative pentose phosphate pathway, and reduction of xylitol formation. Strategies on evolutionary engineering were used to improve fermentation kinetics, but the resulting strains were still far from industrial application. Lignocellulosic hydrolysates proved to have different inhibitors derived from lignin and sugar degradation, along with significant amounts of acetic acid, intrinsically related with biomass deconstruction. This, associated with pH, temperature, high ethanol, and other stress fluctuations presented on large scale fermentations led the search for yeasts with more robust backgrounds, like industrial strains, as engineering targets. Some promising yeasts were obtained both from studies of stress tolerance genes and adaptation on hydrolysates. Since fermentation times on mixed-substrate hydrolysates were still not cost-effective, the more selective search for new or engineered sugar transporters for xylose are still the focus of many recent studies. These challenges, as well as under-appreciated process strategies, will be discussed in this review.

  7. PCR on yeast colonies: an improved method for glyco-engineered Saccharomyces cerevisiae

    PubMed Central

    2013-01-01

    Background Saccharomyces cerevisiae is extensively used in bio-industries. However, its genetic engineering to introduce new metabolism pathways can cause unexpected phenotypic alterations. For example, humanisation of the glycosylation pathways is a high priority pharmaceutical industry goal for production of therapeutic glycoproteins in yeast. Genomic modifications can lead to several described physiological changes: biomass yields decrease, temperature sensitivity or cell wall structure modifications. We have observed that deletion of several N-mannosyltransferases in Saccharomyces cerevisiae, results in strains that can no longer be analyzed by classical PCR on yeast colonies. Findings In order to validate our glyco-engineered Saccharomyces cerevisiae strains, we developed a new protocol to carry out PCR directly on genetically modified yeast colonies. A liquid culture phase, combined with the use of a Hot Start DNA polymerase, allows a 3-fold improvement of PCR efficiency. The results obtained are repeatable and independent of the targeted sequence; as such the protocol is well adapted for intensive screening applications. Conclusions The developed protocol enables by-passing of many of the difficulties associated with PCR caused by phenotypic modifications brought about by humanisation of the glycosylation in yeast and allows rapid validation of glyco-engineered Saccharomyces cerevisiae cells. It has the potential to be extended to other yeast strains presenting cell wall structure modifications. PMID:23688076

  8. Xylose Fermentation by Saccharomyces cerevisiae: Challenges and Prospects

    PubMed Central

    Moysés, Danuza Nogueira; Reis, Viviane Castelo Branco; de Almeida, João Ricardo Moreira; de Moraes, Lidia Maria Pepe; Torres, Fernando Araripe Gonçalves

    2016-01-01

    Many years have passed since the first genetically modified Saccharomyces cerevisiae strains capable of fermenting xylose were obtained with the promise of an environmentally sustainable solution for the conversion of the abundant lignocellulosic biomass to ethanol. Several challenges emerged from these first experiences, most of them related to solving redox imbalances, discovering new pathways for xylose utilization, modulation of the expression of genes of the non-oxidative pentose phosphate pathway, and reduction of xylitol formation. Strategies on evolutionary engineering were used to improve fermentation kinetics, but the resulting strains were still far from industrial application. Lignocellulosic hydrolysates proved to have different inhibitors derived from lignin and sugar degradation, along with significant amounts of acetic acid, intrinsically related with biomass deconstruction. This, associated with pH, temperature, high ethanol, and other stress fluctuations presented on large scale fermentations led the search for yeasts with more robust backgrounds, like industrial strains, as engineering targets. Some promising yeasts were obtained both from studies of stress tolerance genes and adaptation on hydrolysates. Since fermentation times on mixed-substrate hydrolysates were still not cost-effective, the more selective search for new or engineered sugar transporters for xylose are still the focus of many recent studies. These challenges, as well as under-appreciated process strategies, will be discussed in this review. PMID:26927067

  9. Genome Analysis of the Biotechnologically Relevant Acidophilic Iron Oxidising Strain JA12 Indicates Phylogenetic and Metabolic Diversity within the Novel Genus “Ferrovum”

    PubMed Central

    Ullrich, Sophie R.; Poehlein, Anja; Tischler, Judith S.; González, Carolina; Ossandon, Francisco J.; Daniel, Rolf; Holmes, David S.; Schlömann, Michael; Mühling, Martin

    2016-01-01

    Background Members of the genus “Ferrovum” are ubiquitously distributed in acid mine drainage (AMD) waters which are characterised by their high metal and sulfate loads. So far isolation and microbiological characterisation have only been successful for the designated type strain “Ferrovum myxofaciens” P3G. Thus, knowledge about physiological characteristics and the phylogeny of the genus “Ferrovum” is extremely scarce. Objective In order to access the wider genetic pool of the genus “Ferrovum” we sequenced the genome of a “Ferrovum”-containing mixed culture and successfully assembled the almost complete genome sequence of the novel “Ferrovum” strain JA12. Phylogeny and Lifestyle The genome-based phylogenetic analysis indicates that strain JA12 and the type strain represent two distinct “Ferrovum” species. “Ferrovum” strain JA12 is characterised by an unusually small genome in comparison to the type strain and other iron oxidising bacteria. The prediction of nutrient assimilation pathways suggests that “Ferrovum” strain JA12 maintains a chemolithoautotrophic lifestyle utilising carbon dioxide and bicarbonate, ammonium and urea, sulfate, phosphate and ferrous iron as carbon, nitrogen, sulfur, phosphorous and energy sources, respectively. Unique Metabolic Features The potential utilisation of urea by “Ferrovum” strain JA12 is moreover remarkable since it may furthermore represent a strategy among extreme acidophiles to cope with the acidic environment. Unlike other acidophilic chemolithoautotrophs “Ferrovum” strain JA12 exhibits a complete tricarboxylic acid cycle, a metabolic feature shared with the closer related neutrophilic iron oxidisers among the Betaproteobacteria including Sideroxydans lithotrophicus and Thiobacillus denitrificans. Furthermore, the absence of characteristic redox proteins involved in iron oxidation in the well-studied acidophiles Acidithiobacillus ferrooxidans (rusticyanin) and Acidithiobacillus ferrivorans (iron oxidase) indicates the existence of a modified pathway in “Ferrovum” strain JA12. Therefore, the results of the present study extend our understanding of the genus “Ferrovum” and provide a comprehensive framework for future comparative genome and metagenome studies. PMID:26808278

  10. The strains recommended for use in the bacterial reverse mutation test (OECD guideline 471) can be certified as non-genetically modified organisms.

    PubMed

    Sugiyama, Kei-Ichi; Yamada, Masami; Awogi, Takumi; Hakura, Atsushi

    2016-01-01

    The bacterial reverse mutation test, commonly called Ames test, is used worldwide. In Japan, the genetically modified organisms (GMOs) are regulated under the Cartagena Domestic Law, and organisms obtained by self-cloning and/or natural occurrence would be exempted from the law case by case. The strains of Salmonella typhimurium and Escherichia coli recommended for use in the bacterial reverse mutation test (OECD guideline 471), have been considered as non-GMOs because they can be constructed by self-cloning or naturally occurring bacterial strains, or do not disturb the biological diversity. The present article explains the reasons why these tester strains should be classified as non-GMOs.

  11. Myostatin propeptide mutation of the hypermuscular Compact mice decreases the formation of myostatin and improves insulin sensitivity.

    PubMed

    Kocsis, Tamas; Trencsenyi, Gyorgy; Szabo, Kitti; Baan, Julia Aliz; Muller, Geza; Mendler, Luca; Garai, Ildiko; Reinauer, Hans; Deak, Ferenc; Dux, Laszlo; Keller-Pinter, Aniko

    2017-03-01

    The TGFβ family member myostatin (growth/differentiation factor-8) is a negative regulator of skeletal muscle growth. The hypermuscular Compact mice carry the 12-bp Mstn(Cmpt-dl1Abc) deletion in the sequence encoding the propeptide region of the precursor promyostatin, and additional modifier genes of the Compact genetic background contribute to determine the full expression of the phenotype. In this study, by using mice strains carrying mutant or wild-type myostatin alleles with the Compact genetic background and nonmutant myostatin with the wild-type background, we studied separately the effect of the Mstn(Cmpt-dl1Abc) mutation or the Compact genetic background on morphology, metabolism, and signaling. We show that both the Compact myostatin mutation and Compact genetic background account for determination of skeletal muscle size. Despite the increased musculature of Compact s, the absolute size of heart and kidney is not influenced by myostatin mutation; however, the Compact genetic background increases them. Both Compact myostatin and genetic background exhibit systemic metabolic effects. The Compact mutation decreases adiposity and improves whole body glucose uptake, insulin sensitivity, and 18 FDG uptake of skeletal muscle and white adipose tissue, whereas the Compact genetic background has the opposite effect. Importantly, the mutation does not prevent the formation of mature myostatin; however, a decrease in myostatin level was observed, leading to altered activation of Smad2, Smad1/5/8, and Akt, and an increased level of p-AS160, a Rab-GTPase-activating protein responsible for GLUT4 translocation. Based on our analysis, the Compact genetic background strengthens the effect of myostatin mutation on muscle mass, but those can compensate for each other when systemic metabolic effects are compared. Copyright © 2017 the American Physiological Society.

  12. Does race/ethnicity moderate the association between job strain and leisure time physical activity?

    PubMed

    Bennett, Gary G; Wolin, Kathleen Y; Avrunin, Jill S; Stoddard, Anne M; Sorensen, Glorian; Barbeau, Elizabeth; Emmons, Karen M

    2006-08-01

    Racial/ethnic minorities report myriad barriers to regular leisure time physical activity (LTPA), including the stress and fatigue resulting from their occupational activities. We sought to investigate whether an association exists between job strain and LTPA, and whether it is modified by race or ethnicity. Data were collected from 1,740 adults employed in 26 small manufacturing businesses in eastern Massachusetts. LTPA and job strain data were self-reported. Adjusted mean hours of LTPA per week are reported. In age and gender adjusted analyses, reports of job strain were associated with LTPA. There was a significant interaction between job strain and race or ethnicity (p = .04). Whites experiencing job strain reported 1 less hr of LTPA per week compared to Whites not reporting job strain. Collectively, racial/ethnic minorities reporting job strain exhibited comparatively higher levels of LTPA compared to their counterparts with no job strain, although patterns for individual groups did not significantly differ. Job strain was associated with LTPA in a lower income, multiethnic population of healthy adult men and women. The association between job strain and LTPA was modified by race or ethnicity, highlighting the importance of investigating the differential effects of psychosocial occupational factors on LTPA levels by race or ethnicity.

  13. Modified PEHPS medium as an alternative for the in vitro culture of Giardia lamblia.

    PubMed

    Vargas-Villarreal, Javier; Mata-Cárdenas, Benito D; Hernández-García, Magda E; Garza-González, Jesús N; De La Garza-Salinas, Laura H; González-Salazar, Francisco

    2014-01-01

    Commercial culture media present interlot variations in biological activity. We have previously designed a homemade and economic culture medium, PEHPS medium, for the axenic cultivation of Entamoeba histolytica and Trichomonas vaginalis. Trophozoites of amoebae and trichomonads grow well in this medium. Furthermore, the medium is stable for several months when stored frozen or refrigerated. The objective of this work was to modify PEHPS medium to support the in vitro growth of Giardia lamblia. Inocula of 5 × 10(3) trophozoites/mL of G. lamblia were incubated at 36.5°C in modified PEHPS or TYI-S-33 medium. Then, the growths of the three Giardia strains in both media were compared. The logarithmic growth phase lasted 72 h; the mean yield of the strains ranged from 10.06 to 11.43 × 10(5) Giardia trophozoites/mL, and the range of duplication time in the three strains was from 5.67 to 6.06 in modified PEHPS medium. These growth characteristics were not significantly different from those obtained with TYI-S-33 medium. We conclude that modified PEHPS medium might be used for the axenic cultivation of G. lamblia.

  14. Financial strain and cancer risk behaviors among African Americans.

    PubMed

    Advani, Pragati S; Reitzel, Lorraine R; Nguyen, Nga T; Fisher, Felicia D; Savoy, Elaine J; Cuevas, Adolfo G; Wetter, David W; McNeill, Lorna H

    2014-06-01

    African Americans suffer disproportionately from the adverse consequences of behavioral risk factors for cancer relative to other ethnic groups. Recent studies have assessed how financial strain might uniquely contribute to engagement in modifiable behavioral risk factors for cancer, but not among African Americans. The current study examined associations between financial strain and modifiable cancer risk factors (smoking, at-risk alcohol use, overweight/obesity, insufficient physical activity, inadequate fruit and vegetable intake, and multiple risk factors) among 1,278 African American adults (age, 46.5 ± 12.6 years; 77% female) and explored potential mediators (stress and depressive symptoms) of those associations. Logistic regression models were used to examine associations between financial strain and cancer risk factors. Analyses were adjusted for age, sex, partner status, income, educational level, and employment status. Analyses involving overweight/obesity status additionally controlled for fruit and vegetable intake and physical activity. Nonparametric bootstrapping procedures were used to assess mediation. Greater financial strain was associated with greater odds of insufficient physical activity (P < 0.003) and smoking (P = 0.005) and was positively associated with the total number of cancer risk factors (P < 0.0001). There was a significant indirect effect of both stress and depressive symptoms on the relations of financial strain with physical inactivity and multiple risk factors, respectively. Future interventions aimed at reducing cancer disparities should focus on African Americans experiencing higher financial strain while addressing their stress and depressive symptoms. Longitudinal studies are needed to assess the temporal and causal relations between financial strain and modifiable behavioral cancer risk factors among African Americans. ©2014 American Association for Cancer Research.

  15. Fatigue properties of JIS H3300 C1220 copper for strain life prediction

    NASA Astrophysics Data System (ADS)

    Harun, Muhammad Faiz; Mohammad, Roslina

    2018-05-01

    The existing methods for estimating strain life parameters are dependent on the material's monotonic tensile properties. However, a few of these methods yield quite complicated expressions for calculating fatigue parameters, and are specific to certain groups of materials only. The Universal Slopes method, Modified Universal Slopes method, Uniform Material Law, the Hardness method, and Medians method are a few existing methods for predicting strain-life fatigue based on monotonic tensile material properties and hardness of material. In the present study, nine methods for estimating fatigue life and properties are applied on JIS H3300 C1220 copper to determine the best methods for strain life estimation of this ductile material. Experimental strain-life curves are compared to estimations obtained using each method. Muralidharan-Manson's Modified Universal Slopes method and Bäumel-Seeger's method for unalloyed and low-alloy steels are found to yield batter accuracy in estimating fatigue life with a deviation of less than 25%. However, the prediction of both methods only yield much better accuracy for a cycle of less than 1000 or for strain amplitudes of more than 1% and less than 6%. Manson's Original Universal Slopes method and Ong's Modified Four-Point Correlation method are found to predict the strain-life fatigue of copper with better accuracy for a high number of cycles of strain amplitudes of less than 1%. The differences between mechanical behavior during monotonic and cyclic loading and the complexity in deciding the coefficient in an equation are probably the reason for the lack of a reliable method for estimating fatigue behavior using the monotonic properties of a group of materials. It is therefore suggested that a differential approach and new expressions be developed to estimate the strain-life fatigue parameters for ductile materials such as copper.

  16. Effects of Adiabatic Heating on the High Strain Rate Deformation of Polymer Matrix Composites

    NASA Technical Reports Server (NTRS)

    Sorini, Chris; Chattopadhyay, Aditi; Goldberg, Robert K.

    2017-01-01

    Polymer matrix composites (PMCs) are increasingly being used in aerospace structures that are expected to experience complex dynamic loading conditions throughout their lifetime. As such, a detailed understanding of the high strain rate behavior of the constituents, particularly the strain rate, temperature, and pressure dependent polymer matrix, is paramount. In this paper, preliminary efforts in modeling experimentally observed temperature rises due to plastic deformation in PMCs subjected to dynamic loading are presented. To this end, an existing isothermal viscoplastic polymer constitutive formulation is extended to model adiabatic conditions by incorporating temperature dependent elastic properties and modifying the components of the inelastic strain rate tensor to explicitly depend on temperature. It is demonstrated that the modified polymer constitutive model is capable of capturing strain rate and temperature dependent yield as well as thermal softening associated with the conversion of plastic work to heat at high rates of strain. The modified constitutive model is then embedded within a strength of materials based micromechanics framework to investigate the manifestation of matrix thermal softening, due to the conversion of plastic work to heat, on the high strain rate response of a T700Epon 862 (T700E862) unidirectional composite. Adiabatic model predictions for high strain rate composite longitudinal tensile, transverse tensile, and in-plane shear loading are presented. Results show a substantial deviation from isothermal conditions; significant thermal softening is observed for matrix dominated deformation modes (transverse tension and in-plane shear), highlighting the importance of accounting for the conversion of plastic work to heat in the polymer matrix in the high strain rate analysis of PMC structures.

  17. A new Tla region antigen Qa-11, similar to Qa-2 and associated with B-type beta 2-microglobulin.

    PubMed

    van de Meugheuvel, W; van Seventer, G; Demant, P

    1985-04-01

    A new antigen, Qa-11, is detected as a 40,000 dalton band in the SDS-PAGE of immunoprecipitates of radiolabeled lymphocyte membrane preparations. In C57BL H-2 congenic strains, its presence is controlled by a gene in the Tla region. In strains with genetic background other than C57BL it is not expressed. Tests with recombinant inbred strains and with H-3 congenic strains show that, in addition to the Tla region, a gene linked to or identical with the beta 2-microglobulin-b-allele is required for the expression of Qa-11 as well. The mobility of the Qa-11 antigen in SDS-PAGE and in isoelectrofocusing is the same as that of Qa-2 antigen. The Cleveland peptide maps of Qa-2 and Qa-11 are identical as well. This finding, that the Tla region controlled Qa-11 antigen is structurally very similar to the Qa-2 antigen, contrasts with the fact that Tla region products do not react with anti-Qa-2 sera. This paradox could be explained by a separate Qa-11 region between Qa-2 and Tla. Alternatively, it is possible that the Qa-11 antigen is the result of the action of a modifying gene in the Tla region upon a Qa-2 gene product, or that the structural gene for Qa-11 is located in the Qa-2 region and a Tla region gene controls its expression.

  18. Study on elevated-temperature flow behavior of Ni-Cr-Mo-B ultra-heavy-plate steel via experiment and modelling

    NASA Astrophysics Data System (ADS)

    Gao, Zhi-yu; Kang, Yu; Li, Yan-shuai; Meng, Chao; Pan, Tao

    2018-04-01

    Elevated-temperature flow behavior of a novel Ni-Cr-Mo-B ultra-heavy-plate steel was investigated by conducting hot compressive deformation tests on a Gleeble-3800 thermo-mechanical simulator at a temperature range of 1123 K–1423 K with a strain rate range from 0.01 s‑1 to10 s‑1 and a height reduction of 70%. Based on the experimental results, classic strain-compensated Arrhenius-type, a new revised strain-compensated Arrhenius-type and classic modified Johnson-Cook constitutive models were developed for predicting the high-temperature deformation behavior of the steel. The predictability of these models were comparatively evaluated in terms of statistical parameters including correlation coefficient (R), average absolute relative error (AARE), average root mean square error (RMSE), normalized mean bias error (NMBE) and relative error. The statistical results indicate that the new revised strain-compensated Arrhenius-type model could give prediction of elevated-temperature flow stress for the steel accurately under the entire process conditions. However, the predicted values by the classic modified Johnson-Cook model could not agree well with the experimental values, and the classic strain-compensated Arrhenius-type model could track the deformation behavior more accurately compared with the modified Johnson-Cook model, but less accurately with the new revised strain-compensated Arrhenius-type model. In addition, reasons of differences in predictability of these models were discussed in detail.

  19. Conserving, Distributing and Managing Genetically Modified Mouse Lines by Sperm Cryopreservation

    PubMed Central

    Farley, Jane S.; Taft, Robert A.

    2008-01-01

    Background Sperm from C57BL/6 mice are difficult to cryopreserve and recover. Yet, the majority of genetically modified (GM) lines are maintained on this genetic background. Methodology/Principal Findings Reported here is the development of an easily implemented method that consistently yields fertilization rates of 70±5% with this strain. This six-fold increase is achieved by collecting sperm from the vas deferens and epididymis into a cryoprotective medium of 18% raffinose (w/v), 3% skim milk (w/v) and 477 µM monothioglycerol. The sperm suspension is loaded into 0.25 mL French straws and cooled at 37±1°C/min before being plunged and then stored in LN2. Subsequent to storage, the sperm are warmed at 2,232±162°C/min and incubated in in vitro fertilization media for an hour prior to the addition of oocyte cumulus masses from superovulated females. Sperm from 735 GM mouse lines on 12 common genetic backgrounds including C57BL/6J, BALB/cJ, 129S1/SvImJ, FVB/NJ and NOD/ShiLtJ were cryopreserved and recovered. C57BL/6J and BALB/cByJ fertilization rates, using frozen sperm, were slightly reduced compared to rates involving fresh sperm; fertilization rates using fresh or frozen sperm were equivalent in all other lines. Developmental capacity of embryos produced using cryopreserved sperm was equivalent, or superior to, cryopreserved IVF-derived embryos. Conclusions/Significance Combined, these results demonstrate the broad applicability of our approach as an economical and efficient option for archiving and distributing mice. PMID:18665210

  20. Dynamic shear rheology of colloidal suspensions of surface-modified silica nanoparticles in PEG

    NASA Astrophysics Data System (ADS)

    Swarna; Pattanayek, Sudip Kumar; Ghosh, Anup Kumar

    2018-03-01

    The present work illustrates the effect of surface modification of silica nanoparticles (500 nm) with 3-(glycidoxypropyl)trimethoxy silane which was carried out at different reaction times. The suspensions prepared from modified and unmodified silica nanoparticles were evaluated for their shear rate-dependent viscosity and strain-frequency-dependent modulus. The linear viscoelastic moduli, viz., storage modulus and loss modulus, were compared with those of nonlinear moduli. The shear-thickened suspensions displayed strain thinning at low-frequency smaller strains and a strong strain overshoot at higher strains, characteristics of a continuous shear thickening fluids. The shear-thinned suspension, conversely, exhibited a strong elastic dominance at smaller strains, but at higher strains, its strain softened observed in the steady shear viscosity plot indicating characteristics of yielding material. Considering higher order harmonic components, the decomposed elastic and viscous stress revealed a pronounced elastic response up to 10% strain and a high viscous damping at larger strains. The current work is one of a kind in demonstrating the effect of silica surface functionalization on the linear and nonlinear viscoelasticity of suspensions showing a unique rheological fingerprint. The suspensions can thus be predicted through rheological studies for their applicability in energy absorbing and damping materials with respect to their mechanical properties.

  1. A curly-tail modifier locus, mct1, on mouse chromosome 17

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

    Letts, V.A.; Schork, N.J.; Frankel, W.N.

    1995-10-10

    The major gene for neural tube defects, ct, in the curly-tail (CT) mouse strain was mapped previously to mouse chromosome 4 by combining linkage data from several backcrosses. The penetrance of the neural tube trait, already incomplete in the CT strain, was further reduced in several of these backcrosses, suggesting the existence of recessive modifiers or strain-specific susceptibility alleles. Here we describe the mapping of a curly-tail modifier locus, mct1, to chromosome 17 in moderate and low penetrance crosses of CT with BALB/cByJ and Mus spretus. No effect of mct1 was seen in a higher penetrance cross with the BXD-8/Tymore » strain, confirming that ct is the major gene in the model. Homozygosity at both ct and mct1 loci was sufficient to account for all of the affected individuals in the BALB/cByJ cross and most of the affected individuals in the M. spretus cross and was the preferred model overall. No evidence was found for epistatic interaction between ct and mct1. 30 refs., 2 figs., 3 tabs.« less

  2. Characterization of Extracellular Polymeric Substances Produced by Pseudomonas fragi Under Air and Modified Atmosphere Packaging.

    PubMed

    Wang, Guang-Yu; Ma, Fang; Wang, Hu-Hu; Xu, Xing-Lian; Zhou, Guang-Hong

    2017-09-01

    Extracellular polymeric substances (EPS) play an important role in bacterial biochemical properties. The characteristics of EPS from 2 strains of Pseudomonas fragi cultured in meat aerobically (control) and in modified atmosphere packaging (MAP) were studied. The amount and components of EPS, the surface properties, and the effect on biofilm formation of several spoilage organisms were evaluated. The results showed that MAP inhibited the growth of the P. fragi strains. Compared with the control, more loose and less bound EPS (containing protein and carbohydrate) were produced by P. fragi in MAP samples. MAP also caused increased cell autoaggregation and surface hydrophobicity. After the removal of the EPS, the surface property changes were strain-dependent, suggesting that membrane compositions were also changed. In addition, the EPS displayed significant antibiofilm activity on Pseudomonas fluorescens and Serratia liquefaciens. In conclusion, P. fragi strains not only modified the amount, components, and surface properties of EPS but also changed the cell membrane compositions to adapt to MAP stress. Moreover, EPS may play an important role in microbial community competitions. © 2017 Institute of Food Technologists®.

  3. Tyrosinase is the modifier of retinoschisis in mice.

    PubMed

    Johnson, Britt A; Cole, Brian S; Geisert, Eldon E; Ikeda, Sakae; Ikeda, Akihiro

    2010-12-01

    X-linked retinoschisis (XLRS) is a form of macular degeneration with a juvenile onset. This disease is caused by mutations in the retinoschisin (RS1) gene. The major clinical pathologies of this disease include splitting of the retina (schisis) and a loss in synaptic transmission. Human XLRS patients display a broad range in phenotypic severity, even among family members with the same mutation. This variation suggests the existence of genetic modifiers that may contribute to disease severity. Previously, we reported the identification of a modifier locus, named Mor1, which affects severity of schisis in a mouse model of XLRS (the Rs1tmgc1 mouse). Homozygosity for the protective AKR allele of Mor1 restores cell adhesion in Rs1tmgc1 mice. Here, we report our study to identify the Mor1 gene. Through collecting recombinant mice followed by progeny testing, we have localized Mor1 to a 4.4-Mb region on chromosome 7. In this genetic region, the AKR strain is known to carry a mutation in the tyrosinase (Tyr) gene. We observed that the schisis phenotype caused by the Rs1 mutation is rescued by a Tyr mutation in the C57BL/6J genetic background, strongly suggesting that Tyr is the Mor1 gene.

  4. A Modified R-Type Bacteriocin Specifically Targeting Clostridium difficile Prevents Colonization of Mice without Affecting Gut Microbiota Diversity

    PubMed Central

    Gebhart, Dana; Lok, Stephen; Clare, Simon; Tomas, Myreen; Stares, Mark; Scholl, Dean; Donskey, Curtis J.; Lawley, Trevor D.

    2015-01-01

    ABSTRACT Clostridium difficile is a leading cause of nosocomial infections worldwide and has become an urgent public health threat requiring immediate attention. Epidemic lineages of the BI/NAP1/027 strain type have emerged and spread through health care systems across the globe over the past decade. Limiting person-to-person transmission and eradicating C. difficile, especially the BI/NAP1/027 strain type, from health care facilities are difficult due to the abundant shedding of spores that are impervious to most interventions. Effective prophylaxis for C. difficile infection (CDI) is lacking. We have genetically modified a contractile R-type bacteriocin (“diffocin”) from C. difficile strain CD4 to kill BI/NAP1/027-type strains for this purpose. The natural receptor binding protein (RBP) responsible for diffocin targeting was replaced with a newly discovered RBP identified within a prophage of a BI/NAP1/027-type target strain by genome mining. The resulting modified diffocins (a.k.a. Avidocin-CDs), Av-CD291.1 and Av-CD291.2, were stable and killed all 16 tested BI/NAP1/027-type strains. Av-CD291.2 administered in drinking water survived passage through the mouse gastrointestinal (GI) tract, did not detectably alter the mouse gut microbiota or disrupt natural colonization resistance to C. difficile or the vancomycin-resistant Enterococcus faecium (VREF), and prevented antibiotic-induced colonization of mice inoculated with BI/NAP1/027-type spores. Given the high incidence and virulence of the pathogen, preventing colonization by BI/NAP1/027-type strains and limiting their transmission could significantly reduce the occurrence of the most severe CDIs. This modified diffocin represents a prototype of an Avidocin-CD platform capable of producing targetable, precision anti-C. difficile agents that can prevent and potentially treat CDIs without disrupting protective indigenous microbiota. PMID:25805733

  5. Rock Fracture Toughness Under Mode II Loading: A Theoretical Model Based on Local Strain Energy Density

    NASA Astrophysics Data System (ADS)

    Rashidi Moghaddam, M.; Ayatollahi, M. R.; Berto, F.

    2018-01-01

    The values of mode II fracture toughness reported in the literature for several rocks are studied theoretically by using a modified criterion based on strain energy density averaged over a control volume around the crack tip. The modified criterion takes into account the effect of T-stress in addition to the singular terms of stresses/strains. The experimental results are related to mode II fracture tests performed on the semicircular bend and Brazilian disk specimens. There are good agreements between theoretical predictions using the generalized averaged strain energy density criterion and the experimental results. The theoretical results reveal that the value of mode II fracture toughness is affected by the size of control volume around the crack tip and also the magnitude and sign of T-stress.

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

    Sardi, Maria; Rovinskiy, Nikolay; Zhang, Yaoping

    We report a major obstacle to sustainable lignocellulosic biofuel production is microbe inhibition by the combinatorial stresses in pretreated plant hydrolysate. Chemical biomass pretreatment releases a suite of toxins that interact with other stressors, including high osmolarity and temperature, which together can have poorly understood synergistic effects on cells. Improving tolerance in industrial strains has been hindered, in part because the mechanisms of tolerance reported in the literature often fail to recapitulate in other strain backgrounds. Here, we explored and then exploited variations in stress tolerance, toxin-induced transcriptomic responses, and fitness effects of gene overexpression in different Saccharomyces cerevisiae (yeast)more » strains to identify genes and processes linked to tolerance of hydrolysate stressors. Using six different S. cerevisiae strains that together maximized phenotypic and genetic diversity, first we explored transcriptomic differences between resistant and sensitive strains to identify common and strain-specific responses. This comparative analysis implicated primary cellular targets of hydrolysate toxins, secondary effects of defective defense strategies, and mechanisms of tolerance. Dissecting the responses to individual hydrolysate components across strains pointed to synergistic interactions between osmolarity, pH, hydrolysate toxins, and nutrient composition. By characterizing the effects of high-copy gene overexpression in three different strains, we revealed the breadth of the background-specific effects of gene fitness contributions in synthetic hydrolysate. Lastly, our approach identified new genes for engineering improved stress tolerance in diverse strains while illuminating the effects of genetic background on molecular mechanisms.« less

  7. The impact of pneumolysin on the macrophage response to Streptococcus pneumoniae is strain-dependent.

    PubMed

    Harvey, Richard M; Hughes, Catherine E; Paton, Adrienne W; Trappetti, Claudia; Tweten, Rodney K; Paton, James C

    2014-01-01

    Streptococcus pneumoniae is the world's leading cause of pneumonia, bacteremia, meningitis and otitis media. A major pneumococcal virulence factor is the cholesterol-dependent cytolysin, which has the defining property of forming pores in cholesterol-containing membranes. In recent times a clinically significant and internationally successful serotype 1 ST306 clone has been found to express a non-cytolytic variant of Ply (Ply306). However, while the pneumococcus is a naturally transformable organism, strains of the ST306 clonal group have to date been virtually impossible to transform, severely restricting efforts to understand the role of non-cytolytic Ply in the success of this clone. In this study isogenic Ply mutants were constructed in the D39 background and for the first time in the ST306 background (A0229467) to enable direct comparisons between Ply variants for their impact on the immune response in a macrophage-like cell line. Strains that expressed cytolytic Ply were found to induce a significant increase in IL-1β release from macrophage-like cells compared to the non-cytolytic and Ply-deficient strains in a background-independent manner, confirming the requirement for pore formation in the Ply-dependent activation of the NLRP3 inflammasome. However, cytolytic activity in the D39 background was found to induce increased expression of the genes encoding GM-CSF (CSF2), p19 subunit of IL-23 (IL23A) and IFNβ (IFNB1) compared to non-cytolytic and Ply-deficient D39 mutants, but had no effect in the A0229467 background. The impact of Ply on the immune response to the pneumococcus is highly dependent on the strain background, thus emphasising the importance of the interaction between specific virulence factors and other components of the genetic background of this organism.

  8. The Impact of Pneumolysin on the Macrophage Response to Streptococcus pneumoniae is Strain-Dependent

    PubMed Central

    Paton, Adrienne W.; Trappetti, Claudia; Tweten, Rodney K.; Paton, James C.

    2014-01-01

    Streptococcus pneumoniae is the world's leading cause of pneumonia, bacteremia, meningitis and otitis media. A major pneumococcal virulence factor is the cholesterol-dependent cytolysin, which has the defining property of forming pores in cholesterol-containing membranes. In recent times a clinically significant and internationally successful serotype 1 ST306 clone has been found to express a non-cytolytic variant of Ply (Ply306). However, while the pneumococcus is a naturally transformable organism, strains of the ST306 clonal group have to date been virtually impossible to transform, severely restricting efforts to understand the role of non-cytolytic Ply in the success of this clone. In this study isogenic Ply mutants were constructed in the D39 background and for the first time in the ST306 background (A0229467) to enable direct comparisons between Ply variants for their impact on the immune response in a macrophage-like cell line. Strains that expressed cytolytic Ply were found to induce a significant increase in IL-1β release from macrophage-like cells compared to the non-cytolytic and Ply-deficient strains in a background-independent manner, confirming the requirement for pore formation in the Ply-dependent activation of the NLRP3 inflammasome. However, cytolytic activity in the D39 background was found to induce increased expression of the genes encoding GM-CSF (CSF2), p19 subunit of IL-23 (IL23A) and IFNβ (IFNB1) compared to non-cytolytic and Ply-deficient D39 mutants, but had no effect in the A0229467 background. The impact of Ply on the immune response to the pneumococcus is highly dependent on the strain background, thus emphasising the importance of the interaction between specific virulence factors and other components of the genetic background of this organism. PMID:25105894

  9. evaluation of the bioconversion of genetically modified switchgrass using simultaneous saccharification and fermentation ans a consolidated bioprocessing approach

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

    Yee, Kelsey L; Rodriguez, Jr., Miguel; Tschaplinski, Timothy J

    2012-01-01

    Abstract Background: The inherent recalcitrance of lignocellulosic biomass is one of the major economic hurdles for the production of fuels and chemicals from biomass. Additionally, lignin is recognized as having a negative impact on enzymatic hydrolysis of biomass, and as a result much interest has been placed on modifying the lignin pathway to improve bioconversion of lignocellulosic feedstocks. Results: Previous results showed down-regulation of the caffeic acid 3-O-methyl transferase (COMT) gene in the lignin pathway yielded switchgrass (Panicum virgatum) that was more susceptible to bioconversion after dilute acid pretreatment. Here we examined the response of these plant lines to mildermore » pretreatment conditions with yeast-based SSF, CBP with Clostridium thermocellum, and fermentations with the cellulolytic extreme thermophiles, Caldicellulosiruptor bescii and Caldicellulosiruptor obsidiansis. Unlike the S. cerevisiae SSF conversions, fermentations of pretreated down-regulated COMT transgenic switchgrass with C. thermocellum showed an apparent inhibition of fermentation not observed in the wild-type switchgrass. This inhibition can be eliminated by hot water extraction of the pretreated biomass which resulted in superior conversion yield with transgenic versus wild-type switchgrass for C. thermocellum, also exceeding the yeast-based SSF yield. Further fermentation evaluation of the transgenic switchgrass indicated differential inhibition for the Caldicellulosiruptor strains, which could not be rectified by additional processing conditions. Gas chromatography-mass spectrometry metabolite profiling was used to examine the fermentation broth to elucidate the relative abundance of lignin derived aromatic compounds. The types and abundance of fermentation-derived lignin constituents varied between C. thermocellum and each of the Caldicellulosiruptor strains. Conclusions: The down-regulation of the COMT gene improves the bioconversion of switchgrass relative to the wild-type regardless of the pretreatment condition or fermentation microorganism. However, bacterial fermentations demonstrated strain-dependent sensitivity to the COMT transgenic biomass, likely due to additional soluble lignin pathway-derived constituents resulting from the COMT gene disruption. Removal of these inhibitory constituents permitted completion of fermentation by C. thermocellum, but not by the Caldicellulosiruptor strains. The reason for this difference needs to be explored further.« less

  10. The effects of host age, host nuclear background and temperature on phenotypic effects of the virulent Wolbachia strain popcorn in Drosophila melanogaster.

    PubMed

    Reynolds, K Tracy; Thomson, Linda J; Hoffmann, Ary A

    2003-07-01

    Because of their obligate endosymbiotic nature, Wolbachia strains by necessity are defined by their phenotypic effects upon their host. Nevertheless, studies on the influence of host background and environmental conditions upon the manifestation of Wolbachia effects are relatively uncommon. Here we examine the behavior of the overreplicating Wolbachia strain popcorn in four different Drosophila melanogaster backgrounds at two temperatures. Unlike other strains of Wolbachia in Drosophila, popcorn has a major fitness impact upon its hosts. The rapid proliferation of popcorn causes cells to rupture, resulting in the premature death of adult hosts. Apart from this effect, we found that popcorn delayed development time, and host background influenced both this trait and the rate of mortality associated with infection. Temperature influenced the impact of popcorn upon host mortality, with no reduction in life span occurring in flies reared at 19 degrees. No effect upon fecundity was found. Contrary to earlier reports, popcorn induced high levels of incompatibility when young males were used in tests, and CI levels declined rapidly with male age. The population dynamics of popcorn-type infections will therefore depend on environmental temperature, host background, and the age structure of the population.

  11. Numerical analysis of standard and modified osteosynthesis in long bone fractures treatment.

    PubMed

    Sisljagić, Vladimir; Jovanović, Savo; Mrcela, Tomislav; Radić, Radivoje; Selthofer, Robert; Mrcela, Milanka

    2010-03-01

    The fundamental problem in osteoporotic fracture treatment is significant decrease in bone mass and bone tissue density resulting in decreased firmness and elasticity of osteoporotic bone. Application of standard implants and standard surgical techniques in osteoporotic bone fracture treatment makes it almost impossible to achieve stable osteosynthesis sufficient for early mobility, verticalization and load. Taking into account the form and the size of the contact surface as well as distribution of forces between the osteosynthetic materials and the bone tissue numerical analysis showed advantages of modified osteosynthesis with bone cement filling in the screw bed. The applied numerical model consisted of three sub-models: 3D model from solid elements, 3D cross section of the contact between the plate and the bone and the part of 3D cross section of the screw head and body. We have reached the conclusion that modified osteosynthesis with bone cement resulted in weaker strain in the part of the plate above the fracture fissure, more even strain on the screws, plate and bone, more even strain distribution along all the screws' bodies, significantly greater strain in the part of the screw head opposite to the fracture fissure, firm connection of the screw head and neck and the plate hole with the whole plate and more even bone strain around the screw.

  12. Engineering industrial Saccharomyces cerevisiae strains for xylose fermentation and comparison for switchgrass conversion.

    PubMed

    Hector, Ronald E; Dien, Bruce S; Cotta, Michael A; Qureshi, Nasib

    2011-09-01

    Saccharomyces' physiology and fermentation-related properties vary broadly among industrial strains used to ferment glucose. How genetic background affects xylose metabolism in recombinant Saccharomyces strains has not been adequately explored. In this study, six industrial strains of varied genetic background were engineered to ferment xylose by stable integration of the xylose reductase, xylitol dehydrogenase, and xylulokinase genes. Aerobic growth rates on xylose were 0.04-0.17 h(-1). Fermentation of xylose and glucose/xylose mixtures also showed a wide range of performance between strains. During xylose fermentation, xylose consumption rates were 0.17-0.31 g/l/h, with ethanol yields 0.18-0.27 g/g. Yields of ethanol and the metabolite xylitol were positively correlated, indicating that all of the strains had downstream limitations to xylose metabolism. The better-performing engineered and parental strains were compared for conversion of alkaline pretreated switchgrass to ethanol. The engineered strains produced 13-17% more ethanol than the parental control strains because of their ability to ferment xylose.

  13. Detection and characterization of multidrug-resistant enterobacteria bearing aminoglycoside-modifying gene in a university hospital at Rio de Janeiro, Brazil, along three decades.

    PubMed

    Dias-Gonçalves, Verônica; Bohrer-Lengruber, Françoise; Oliveira-Fonseca, Bianca; Santos-Pereira, Renata Meirelles; Barbosa de Melo, Luis Dione; Gazos-Lopes, Ulisses; Ribeiro-Bello, Alexandre; Adler-Pereira, José Augusto

    2015-01-01

    Multidrug-resistant Enterobacteriaceae, particularly those resistant to gentamicin, have become one of the most important causes of nosocomial infections. We sought to investigate the presence of genes conferring resistance to aminoglycosides, specially to gentamicin, in Klebsiella pneumoniae and Escherichia coli multidrug-resistant strains isolated from different clinical materials among patients hospitalized in a university hospital in Rio de Janeiro, Brazil. Ten colonization strains and 20 infection strains were evaluated during three decades (1980 to 2010) using selective media containing 8 µg/ml of gentamicin. Thirty strains were tested for antimicrobial susceptibility. Twenty two strains were subjected to plasmid DNA extraction and 12 to hybridization assays using as probe a 1.9 kb plasmid DNA fragment from one of the K. pneumoniae strains isolated from faecal samples. This fragment was sequenced and assigned to the GQ422439 GenBank record. PCR was also performed using oligonucleotides designed for aminoglycoside-modifying enzymes. An accC2 acetylase, besides transposons and insertion sequences, were evidenced. Twenty-four (80%) of the isolates were positive for the aacC2 gene in agreement with antibiotic susceptibility testing profiles, indicating the persistent presence of this gene throughout the three decades. We detected high molecular weight plasmids in 54,5% of the strains. Of the tested strains, 91% showed positive signal in the hybridization assays. A gene codifying for one specific aminoglycoside-modifying enzyme was detected all throughout the three decades. Our data back the adoption of preventive measures, such as a more conscious use of antimicrobial agents in hospital environments, which can contribute to control the dissemination of microorganisms harboring resistance gene plasmids.

  14. Unmet Needs of Caregivers of Patients Referred to a Dementia Care Program

    PubMed Central

    Jennings, Lee A.; Reuben, David B.; Evertson, Leslie Chang; Serrano, Katherine S.; Ercoli, Linda; Grill, Joshua; Chodosh, Joshua; Tan, Zaldy; Wenger, Neil S.

    2014-01-01

    Background/Objectives Caregiver strain and low self-efficacy for managing dementia-related problems are common among those caring for patients with dementia, but the level of unmet need and relation to provider type has not been well characterized. Design Cross-sectional observational cohort Setting Urban academic medical center Participants Caregivers of community-dwelling adults with dementia referred to the program Measurements Caregivers were surveyed and completed the Patient Health Questionnaire (PHQ-9) about themselves, the Modified Caregiver Strain Index, the Neuropsychiatric Inventory Questionnaire, which measures patient symptom severity and related caregiver distress, and a 9-item caregiver self-efficacy scale developed for the study. Results Of 307 patient and caregiver dyads surveyed over a one year period, 32% of caregivers reported confidence in managing dementia-related problems, 19% knew how to access community services to help provide care, and 28% agreed the patient’s provider helped them work through dementia care problems. Thirty-eight percent reported high levels of caregiver strain, and 15% reported moderate to severe depressive symptoms. Caregivers of patients referred by geriatricians more often reported having a healthcare professional to help work through dementia care problems than those referred by internists, family physicians, or other specialists, but self-efficacy did not differ. Low caregiver self-efficacy was associated with higher caregiver strain, more caregiver depressive symptoms, and caring for a patient with more severe behavioral symptoms. Conclusion Most caregivers perceived inadequate support from the patient’s provider in managing dementia-related problems, reported strain, and had low confidence in managing caregiving. New models of care are needed to address the complex care needs of patients with dementia and their caregivers. PMID:25688604

  15. Unraveling Genetic Modifiers in the Gria4 Mouse Model of Absence Epilepsy

    PubMed Central

    Frankel, Wayne N.; Mahaffey, Connie L.; McGarr, Tracy C.; Beyer, Barbara J.; Letts, Verity A.

    2014-01-01

    Absence epilepsy (AE) is a common type of genetic generalized epilepsy (GGE), particularly in children. AE and GGE are complex genetic diseases with few causal variants identified to date. Gria4 deficient mice provide a model of AE, one for which the common laboratory inbred strain C3H/HeJ (HeJ) harbors a natural IAP retrotransposon insertion in Gria4 that reduces its expression 8-fold. Between C3H and non-seizing strains such as C57BL/6, genetic modifiers alter disease severity. Even C3H substrains have surprising variation in the duration and incidence of spike-wave discharges (SWD), the characteristic electroencephalographic feature of absence seizures. Here we discovered extensive IAP retrotransposition in the C3H substrain, and identified a HeJ-private IAP in the Pcnxl2 gene, which encodes a putative multi-transmembrane protein of unknown function, resulting in decreased expression. By creating new Pcnxl2 frameshift alleles using TALEN mutagenesis, we show that Pcnxl2 deficiency is responsible for mitigating the seizure phenotype – making Pcnxl2 the first known modifier gene for absence seizures in any species. This finding gave us a handle on genetic complexity between strains, directing us to use another C3H substrain to map additional modifiers including validation of a Chr 15 locus that profoundly affects the severity of SWD episodes. Together these new findings expand our knowledge of how natural variation modulates seizures, and highlights the feasibility of characterizing and validating modifiers in mouse strains and substrains in the post-genome sequence era. PMID:25010494

  16. Co-Infection Dynamics of a Major Food-Borne Zoonotic Pathogen in Chicken

    PubMed Central

    Skånseng, Beate; Trosvik, Pål; Zimonja, Monika; Johnsen, Gro; Bjerrum, Lotte; Pedersen, Karl; Wallin, Nina; Rudi, Knut

    2007-01-01

    A major bottleneck in understanding zoonotic pathogens has been the analysis of pathogen co-infection dynamics. We have addressed this challenge using a novel direct sequencing approach for pathogen quantification in mixed infections. The major zoonotic food-borne pathogen Campylobacter jejuni, with an important reservoir in the gastrointestinal (GI) tract of chickens, was used as a model. We investigated the co-colonisation dynamics of seven C. jejuni strains in a chicken GI infection trial. The seven strains were isolated from an epidemiological study showing multiple strain infections at the farm level. We analysed time-series data, following the Campylobacter colonisation, as well as the dominant background flora of chickens. Data were collected from the infection at day 16 until the last sampling point at day 36. Chickens with two different background floras were studied, mature (treated with Broilact, which is a product consisting of bacteria from the intestinal flora of healthy hens) and spontaneous. The two treatments resulted in completely different background floras, yet similar Campylobacter colonisation patterns were detected in both groups. This suggests that it is the chicken host and not the background flora that is important in determining the Campylobacter colonisation pattern. Our results showed that mainly two of the seven C. jejuni strains dominated the Campylobacter flora in the chickens, with a shift of the dominating strain during the infection period. We propose a model in which multiple C. jejuni strains can colonise a single host, with the dominant strains being replaced as a consequence of strain-specific immune responses. This model represents a new understanding of C. jejuni epidemiology, with future implications for the development of novel intervention strategies. PMID:18020703

  17. Family resources study: part 1: family resources, family function and caregiver strain in childhood cancer

    PubMed Central

    2011-01-01

    Background Severe illness can disrupt family life, cause family dysfunction, strain resources, and cause caregiver burden. The family's ability to cope with crises depends on their resources. This study sought to assess families of children with cancer in terms of family function-dysfunction, family caregiver strain and the adequacy of family resources using a new family resources assessment instrument. Methods This is a cross-sectional study involving 90 Filipino family caregivers of children undergoing cancer treatment. This used a self-administered questionnaire composed of a new 12-item family resources questionnaire (SCREEM-RES) based on the SCREEM method of analysis, Family APGAR to assess family function-dysfunction; and Modified Caregiver Strain Index to assess strain in caring for the patient. Results More than half of families were either moderately or severely dysfunctional. Close to half of caregivers were either predisposed to strain or experienced severe strain, majority disclosed that their families have inadequate economic resources; many also report inaccessibility to medical help in the community and insufficient educational resources to understand and care for their patients. Resources most often reported as adequate were: family's faith and religion; help from within the family and from health providers. SCREEM-RES showed to be reliable with Cronbach's alpha of 0.80. There is good inter-item correlation between items in each domain: 0.24-0.70. Internal consistency reliability for each domain was also good: 0.40-0.92. Using 2-point scoring system, Cronbach's alpha were slightly lower: full scale (0.70) and for each domain 0.26-.82. Results showed evidence of association between family resources and family function based on the family APGAR but none between family resources and caregiver strain and between family function and caregiver strain. Conclusion Many Filipino families of children with cancer have inadequate resources, especially economic; and are moderately or severely dysfunctional. Many caregivers are predisposed to caregiver strain or are already experiencing severe strain. To provide appropriate care for these families, physicians should regularly assess family function, resources and strain experienced by caregivers. The SCREEM-RES questionnaire used in this study is a helpful and reliable instrument to assess adequacy of family resources. PMID:22040272

  18. Highly efficient gene targeting in Aspergillus oryzae industrial strains under ligD mutation introduced by genome editing: Strain-specific differences in the effects of deleting EcdR, the negative regulator of sclerotia formation.

    PubMed

    Nakamura, Hidetoshi; Katayama, Takuya; Okabe, Tomoya; Iwashita, Kazuhiro; Fujii, Wataru; Kitamoto, Katsuhiko; Maruyama, Jun-Ichi

    2017-07-11

    Numerous strains of Aspergillus oryzae are industrially used for Japanese traditional fermentation and for the production of enzymes and heterologous proteins. In A. oryzae, deletion of the ku70 or ligD genes involved in non-homologous end joining (NHEJ) has allowed high gene targeting efficiency. However, this strategy has been mainly applied under the genetic background of the A. oryzae wild strain RIB40, and it would be laborious to delete the NHEJ genes in many A. oryzae industrial strains, probably due to their low gene targeting efficiency. In the present study, we generated ligD mutants from the A. oryzae industrial strains by employing the CRISPR/Cas9 system, which we previously developed as a genome editing method. Uridine/uracil auxotrophic strains were generated by deletion of the pyrG gene, which was subsequently used as a selective marker. We examined the gene targeting efficiency with the ecdR gene, of which deletion was reported to induce sclerotia formation under the genetic background of the strain RIB40. As expected, the deletion efficiencies were high, around 60~80%, in the ligD mutants of industrial strains. Intriguingly, the effects of the ecdR deletion on sclerotia formation varied depending on the strains, and we found sclerotia-like structures under the background of the industrial strains, which have never been reported to form sclerotia. The present study demonstrates that introducing ligD mutation by genome editing is an effective method allowing high gene targeting efficiency in A. oryzae industrial strains.

  19. Complete In Vitro Life Cycle of Trypanosoma congolense: Development of Genetic Tools

    PubMed Central

    Plazolles, Nicolas; Baltz, Théo

    2010-01-01

    Background Animal African trypanosomosis, a disease mainly caused by the protozoan parasite Trypanosoma congolense, is a major constraint to livestock productivity and has a significant impact in the developing countries of Africa. RNA interference (RNAi) has been used to study gene function and identify drug and vaccine targets in a variety of organisms including trypanosomes. However, trypanosome RNAi studies have mainly been conducted in T. brucei, as a model for human infection, largely ignoring livestock parasites of economical importance such as T. congolense, which displays different pathogenesis profiles. The whole T. congolense life cycle can be completed in vitro, but this attractive model displayed important limitations: (i) genetic tools were currently limited to insect forms and production of modified infectious BSF through differentiation was never achieved, (ii) in vitro differentiation techniques lasted several months, (iii) absence of long-term bloodstream forms (BSF) in vitro culture prevented genomic analyses. Methodology/Principal Findings We optimized culture conditions for each developmental stage and secured the differentiation steps. Specifically, we devised a medium adapted for the strenuous development of stable long-term BSF culture. Using Amaxa nucleofection technology, we greatly improved the transfection rate of the insect form and designed an inducible transgene expression system using the IL3000 reference strain. We tested it by expression of reporter genes and through RNAi. Subsequently, we achieved the complete in vitro life cycle with dramatically shortened time requirements for various wild type and transgenic strains. Finally, we established the use of modified strains for experimental infections and underlined a host adaptation phase requirement. Conclusions/Significance We devised an improved T. congolense model, which offers the opportunity to perform functional genomics analyses throughout the whole life cycle. It represents a very useful tool to understand pathogenesis mechanisms and to study potential therapeutic targets either in vitro or in vivo using a mouse model. PMID:20209144

  20. Influence of genetic background of engineered xylose-fermenting industrial Saccharomyces cerevisiae strains for ethanol production from lignocellulosic hydrolysates

    USDA-ARS?s Scientific Manuscript database

    An industrial ethanol-producing Saccharomyces cerevisiae strain with genes needed for xylose-fermentation integrated into its genome was used to obtain haploids and diploid isogenic strains. The isogenic strains were more effective in metabolizing xylose than their parental strain (p < 0.05) and abl...

  1. Transcriptome profiles of chicken intestinal intraepithelial lymphocytes altered by the intake of a multi-strain direct-fed microbials

    USDA-ARS?s Scientific Manuscript database

    The current study was conducted to investigate the effects of the direct-fed microbials (DFM) including three Bacillus subtilis strains on the modulation of transcriptional profile in chicken intestinal intraepithelial lymphocytes (IEL). The multiple-strain DFM product modified 453 probes from 1,98...

  2. Combinatorial Strategies for Improving Multiple-Stress Resistance in Industrially Relevant Escherichia coli Strains

    PubMed Central

    Herrgård, Markus J.

    2014-01-01

    High-cell-density fermentation for industrial production of chemicals can impose numerous stresses on cells due to high substrate, product, and by-product concentrations; high osmolarity; reactive oxygen species; and elevated temperatures. There is a need to develop platform strains of industrial microorganisms that are more tolerant toward these typical processing conditions. In this study, the growth of six industrially relevant strains of Escherichia coli was characterized under eight stress conditions representative of fed-batch fermentation, and strains W and BL21(DE3) were selected as platforms for transposon (Tn) mutagenesis due to favorable resistance characteristics. Selection experiments, followed by either targeted or genome-wide next-generation-sequencing-based Tn insertion site determination, were performed to identify mutants with improved growth properties under a subset of three stress conditions and two combinations of individual stresses. A subset of the identified loss-of-function mutants were selected for a combinatorial approach, where strains with combinations of two and three gene deletions were systematically constructed and tested for single and multistress resistance. These approaches allowed identification of (i) strain-background-specific stress resistance phenotypes, (ii) novel gene deletion mutants in E. coli that confer single and multistress resistance in a strain-background-dependent manner, and (iii) synergistic effects of multiple gene deletions that confer improved resistance over single deletions. The results of this study underscore the suboptimality and strain-specific variability of the genetic network regulating growth under stressful conditions and suggest that further exploration of the combinatorial gene deletion space in multiple strain backgrounds is needed for optimizing strains for microbial bioprocessing applications. PMID:25085490

  3. Sub-inhibitory stress with essential oil affects enterotoxins production and essential oil susceptibility in Staphylococcus aureus.

    PubMed

    Turchi, Barbara; Mancini, Simone; Pistelli, Luisa; Najar, Basma; Cerri, Domenico; Fratini, Filippo

    2018-03-01

    Fourteen wild strains of Staphylococcus aureus positive for gene sea were tested for enterotoxins production and the minimum inhibitory concentration of Leptospermum scoparium, Origanum majorana, Origanum vulgare, Satureja montana and Thymus vulgaris essential oils (EOs) were determined. After this trial, bacteria stressed with sub-inhibitory concentration of each EO were tested for enterotoxins production by an immunoenzymatic assay and resistance to the same EO. Oregano oil exhibited the highest antibacterial activity followed by manuka and thyme oils. After the exposure to a sub-inhibitory concentration of EOs, strains displayed an increased sensitivity in more than 95% of the cases. After treatment with oregano and marjoram EOs, few strains showed a modified enterotoxins production, while 43% of the strains were no longer able to produce enterotoxins after treatment with manuka EO. The results obtained in this study highlight that exposure to sub-inhibitory concentration of EO modifies strains enterotoxins production and EOs susceptibility profile.

  4. Impact of Saccharomyces cerevisiae metabolites produced during fermentation on bread quality parameters: A review.

    PubMed

    Heitmann, Mareile; Zannini, Emanuele; Arendt, Elke

    2018-05-03

    Although bread making with the use of Baker's yeast has a long tradition in human history, little attention has been paid to the connection between yeast addition and the final bread quality. Nowadays, bakers mainly use different flour additives such as enzymes (amylases, hemicellulases, and proteases) to change and improve dough properties and/or bread quality. Another strategy is the use of modified industrial Baker's yeast. To date, there is no yeast strain used in the baking industry, which is genetically modified, despite some studies demonstrating that the application of recombinant DNA technology is a possibility for improved strains suitable for baking. However, due to the fact that the majority of consumers in Europe highly reject the use of genetically modified microorganisms in the production of food, other strategies to improve bread quality must be investigated. Such a strategy would be a reconsideration of the selection of yeast strains used for the baking process. Next to the common criteria, the requirement for adequate gas production, more attention should be paid on how yeast impacts flavor, shelf life, color, and the nutritional value of baked products, in a similar way to which yeast strains are selected in the wine and brewing industries.

  5. The relevance of inter- and intrastrain differences in mice and rats and their implications for models of seizures and epilepsy.

    PubMed

    Löscher, Wolfgang; Ferland, Russell J; Ferraro, Thomas N

    2017-08-01

    It is becoming increasingly clear that the genetic background of mice and rats, even in inbred strains, can have a profound influence on measures of seizure susceptibility and epilepsy. These differences can be capitalized upon through genetic mapping studies to reveal genes important for seizures and epilepsy. However, strain background and particularly mixed genetic backgrounds of transgenic animals need careful consideration in both the selection of strains and in the interpretation of results and conclusions. For instance, mice with targeted deletions of genes involved in epilepsy can have profoundly disparate phenotypes depending on the background strain. In this review, we discuss findings related to how this genetic heterogeneity has and can be utilized in the epilepsy field to reveal novel insights into seizures and epilepsy. Moreover, we discuss how caution is needed in regards to rodent strain or even animal vendor choice, and how this can significantly influence seizure and epilepsy parameters in unexpected ways. This is particularly critical in decisions regarding the strain of choice used in generating mice with targeted deletions of genes. Finally, we discuss the role of environment (at vendor and/or laboratory) and epigenetic factors for inter- and intrastrain differences and how such differences can affect the expression of seizures and the animals' performance in behavioral tests that often accompany acute and chronic seizure testing. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Isolation and characterization of mutated alcohol oxidases from the yeast Hansenula polymorpha with decreased affinity toward substrates and their use as selective elements of an amperometric biosensor

    PubMed Central

    Dmytruk, Kostyantyn V; Smutok, Oleh V; Ryabova, Olena B; Gayda, Galyna Z; Sibirny, Volodymyr A; Schuhmann, Wolfgang; Gonchar, Mykhailo V; Sibirny, Andriy A

    2007-01-01

    Background Accurate, rapid, and economic on-line analysis of ethanol is very desirable. However, available biosensors achieve saturation at very low ethanol concentrations and thus demand the time and labour consuming procedure of sample dilution. Results Hansenula polymorpha (Pichia angusta) mutant strains resistant to allyl alcohol in methanol medium were selected. Such strains possessed decreased affinity of alcohol oxidase (AOX) towards methanol: the KM values for AOX of wild type and mutant strains CA2 and CA4 are shown to be 0.62, 2.48 and 1.10 mM, respectively, whereas Vmax values are increased or remain unaffected. The mutant AOX alleles from H. polymorpha mutants CA2 and CA4 were isolated and sequenced. Several point mutations in the AOX gene, mostly different between the two mutant alleles, have been identified. Mutant AOX forms were isolated and purified, and some of their biochemical properties were studied. An amperometric biosensor based on the mutated form of AOX from the strain CA2 was constructed and revealed an extended linear response to the target analytes, ethanol and formaldehyde, as compared to the sensor based on the native AOX. Conclusion The described selection methodology opens up the possibility of isolating modified forms of AOX with further decreased affinity toward substrates without reduction of the maximal velocity of reaction. It can help in creation of improved ethanol biosensors with a prolonged linear response towards ethanol in real samples of wines, beers or fermentation liquids. PMID:17567895

  7. The effects of host age, host nuclear background and temperature on phenotypic effects of the virulent Wolbachia strain popcorn in Drosophila melanogaster.

    PubMed Central

    Reynolds, K Tracy; Thomson, Linda J; Hoffmann, Ary A

    2003-01-01

    Because of their obligate endosymbiotic nature, Wolbachia strains by necessity are defined by their phenotypic effects upon their host. Nevertheless, studies on the influence of host background and environmental conditions upon the manifestation of Wolbachia effects are relatively uncommon. Here we examine the behavior of the overreplicating Wolbachia strain popcorn in four different Drosophila melanogaster backgrounds at two temperatures. Unlike other strains of Wolbachia in Drosophila, popcorn has a major fitness impact upon its hosts. The rapid proliferation of popcorn causes cells to rupture, resulting in the premature death of adult hosts. Apart from this effect, we found that popcorn delayed development time, and host background influenced both this trait and the rate of mortality associated with infection. Temperature influenced the impact of popcorn upon host mortality, with no reduction in life span occurring in flies reared at 19 degrees. No effect upon fecundity was found. Contrary to earlier reports, popcorn induced high levels of incompatibility when young males were used in tests, and CI levels declined rapidly with male age. The population dynamics of popcorn-type infections will therefore depend on environmental temperature, host background, and the age structure of the population. PMID:12871912

  8. Note: Modified anvil design for improved reliability in DT-Cup experiments.

    PubMed

    Hunt, Simon A; Dobson, David P

    2017-12-01

    The Deformation T-Cup (DT-Cup) is a modified 6-8 multi-anvil apparatus capable of controlled strain-rate deformation experiments at pressures greater than 18 GPa. Controlled strain-rate deformation was enabled by replacing two of the eight cubic "second-stage" anvils with hexagonal cross section deformation anvils and modifying the "first-stage" wedges. However, with these modifications approximately two-thirds of experiments end with rupture of the hexagonal anvils. By replacing the hexagonal anvils with cubic anvils and, split, deformation wedge extensions, we restore the massive support to the deformation anvils that were inherent in the original multi-anvil design and prevent deformation anvil failure. With the modified parts, the DT-Cup has an experimental success rate that is similar to that of a standard hydrostatic 6-8 multi-anvil apparatus.

  9. Flow Behavior and Constitutive Equation of Ti-6.5Al-2Sn-4Zr-4Mo-1W-0.2Si Titanium Alloy

    NASA Astrophysics Data System (ADS)

    Yang, Xuemei; Guo, Hongzhen; Liang, Houquan; Yao, Zekun; Yuan, Shichong

    2016-04-01

    In order to get a reliable constitutive equation for the finite element simulation, flow behavior of Ti-6.5Al-2Sn-4Zr-4Mo-1W-0.2Si alloy under high temperature was investigated by carrying a series of isothermal compression tests at temperatures of 1153-1293 K and strain rates of 0.01-10.0 s-1 on the Gleeble-1500 simulator. Results showed that the true stress-strain curves exhibited peaks at small strains, after which the flow stress decreased monotonically. Ultimately, the flow curves reached steady state at the strain of 0.6, showing a dynamic flow softening phenomenon. The effects of strain rate, temperature, and strain on the flow behavior were researched by establishing a constitutive equation. The relations among stress exponent, deformation activation energy, and strain were preliminarily discussed by using strain rate sensitivity exponent and dynamic recrystallization kinetics curve. Stress values predicted by the modified constitutive equation showed a good agreement with the experimental ones. The correlation coefficient ( R) and average absolute relative error (AARE) were 98.2% and 4.88%, respectively, which confirmed that the modified constitutive equation could give an accurate estimation of the flow stress for BT25y titanium alloy.

  10. Nutrient depletion modifies cell wall adsorption activity of wine yeast.

    PubMed

    Sidari, R; Caridi, A

    2016-06-01

    Yeast cell wall is a structure that helps yeasts to manage and respond to many environmental stresses. The mannosylphosphorylation is a modification in response to stress that provides the cell wall with negative charges able to bind compounds present in the environment. Phenotypes related to the cell wall modification such as the filamentous growth in Saccharomyces cerevisiae are affected by nutrient depletion. The present work aimed at describing the effect of carbon and/or nitrogen limitation on the aptitude of S. cerevisiae strains to bind coloured polyphenols. Carbon- and nitrogen-rich or deficient media supplemented with grape polyphenols were used to simulate different grape juice conditions-early, mid, 'adjusted' for nitrogen, and late fermentations. In early fermentation condition, the R+G+B values range from 106 (high adsorption, strain Sc1128) to 192 (low adsorption, strain Σ1278b), in mid-fermentation the values range from 111 (high adsorption, strain Sc1321) to 258 (low adsorption, strain Sc2306), in 'adjusted' for nitrogen conditions the values range from 105 (high adsorption, strain Sc1321) to 194 (low adsorption, strain Sc2306) while in late fermentation conditions the values range from 101 (high adsorption, strain Sc384) to 293 (low adsorption, strain Sc2306). The effect of nutrient availability is not univocal for all the strains and the different media tested modified the strains behaviour. In all the media the strains show significant differences. Results demonstrate that wine yeasts decrease/increase their parietal adsorption activity according to the nutrient availability. The wide range of strain variability observed could be useful in selecting wine starters.

  11. Characterization of NDM-1- and OXA-23-producing Acinetobacter baumannii isolates from inanimate surfaces in a hospital environment in Algeria.

    PubMed

    Zenati, K; Touati, A; Bakour, S; Sahli, F; Rolain, J M

    2016-01-01

    Investigation of several outbreaks of multidrug-resistant Acinetobacter baumannii infection has demonstrated that contamination of the inanimate hospital environment could be implicated in the spread of these multidrug-resistant strains. To investigate the occurrence of carbapenem-resistant A. baumannii on inanimate surfaces and possible dissemination in the hospital environment in Algeria as a potential source of infection in humans. A. baumannii strains were isolated from the hospital environment and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Antimicrobial susceptibility was determined using disc diffusion and E-test methods. Carbapenemase activity was detected using microbiological tests, including modified Hodge test, modified Carba NP test, and EDTA test. Carbapenem resistance determinants were studied by polymerase chain reaction (PCR) and sequencing. Clonal relatedness was determined using multi-locus sequence typing (MLST). A total of 67 A. baumannii isolates were obtained from 868 environmental samples and identified by MALDI-TOF MS. Among them, 61 isolates were resistant to imipenem with minimum inhibitory concentration >32 μg/mL and positive by the modified Hodge test and modified Carba NP test. In addition, the activity of carbapenemase was inhibited by EDTA in 32 strains. PCR and sequencing showed the presence of blaOXA-23 gene in 29 strains, and the blaNDM-1 gene in 32 isolates. MLST demonstrated the presence of five types of ST (ST19, ST2, ST85, ST98, and ST115). Our study demonstrated the dissemination of carbapenemase-producing A. baumannii strains recovered from inanimate surfaces in a hospital environment, surrounding patients, healthcare workers and visitors, in Algeria as a potential source for nosocomial infection. Copyright © 2015 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

  12. Zero-magnetic field effect in pathogen bacteria

    NASA Astrophysics Data System (ADS)

    Creanga, D. E.; Poiata, A.; Morariu, V. V.; Tupu, P.

    2004-05-01

    Two lots of Gram-negative bacterial strains were tested for antibiotic drug resistance after exposure to zero-magnetic field. We found that the magneto-sensitive strains represent half of the analyzed samples (three Pseudomonas and five Enterobacter strains), some of them presenting two-three times modified resistance to antibiotic, while others revealed eight or 16 times changed resistance. Pseudomonas strain magnetic sensitivity is revealed better by ampicillin and tetracycline, while Enterobacter strain magnetic sensitivity is revealed better by ampicillin, kanamycin and ofloxacin.

  13. A Murine Hypertrophic Cardiomyopathy Model: The DBA/2J Strain.

    PubMed

    Zhao, Wenyuan; Zhao, Tieqiang; Chen, Yuanjian; Zhao, Fengbo; Gu, Qingqing; Williams, Robert W; Bhattacharya, Syamal K; Lu, Lu; Sun, Yao

    2015-01-01

    Familial hypertrophic cardiomyopathy (HCM) is attributed to mutations in genes that encode for the sarcomere proteins, especially Mybpc3 and Myh7. Genotype-phenotype correlation studies show significant variability in HCM phenotypes among affected individuals with identical causal mutations. Morphological changes and clinical expression of HCM are the result of interactions with modifier genes. With the exceptions of angiotensin converting enzyme, these modifiers have not been identified. Although mouse models have been used to investigate the genetics of many complex diseases, natural murine models for HCM are still lacking. In this study we show that the DBA/2J (D2) strain of mouse has sequence variants in Mybpc3 and Myh7, relative to widely used C57BL/6J (B6) reference strain and the key features of human HCM. Four-month-old of male D2 mice exhibit hallmarks of HCM including increased heart weight and cardiomyocyte size relative to B6 mice, as well as elevated markers for cardiac hypertrophy including β-myosin heavy chain (MHC), atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and skeletal muscle alpha actin (α1-actin). Furthermore, cardiac interstitial fibrosis, another feature of HCM, is also evident in the D2 strain, and is accompanied by up-regulation of type I collagen and α-smooth muscle actin (SMA)-markers of fibrosis. Of great interest, blood pressure and cardiac function are within the normal range in the D2 strain, demonstrating that cardiac hypertrophy and fibrosis are not secondary to hypertension, myocardial infarction, or heart failure. Because D2 and B6 strains have been used to generate a large family of recombinant inbred strains, the BXD cohort, the D2 model can be effectively exploited for in-depth genetic analysis of HCM susceptibility and modifier screens.

  14. Application of Temperature Sensitivities During Iterative Strain-Gage Balance Calibration Analysis

    NASA Technical Reports Server (NTRS)

    Ulbrich, N.

    2011-01-01

    A new method is discussed that may be used to correct wind tunnel strain-gage balance load predictions for the influence of residual temperature effects at the location of the strain-gages. The method was designed for the iterative analysis technique that is used in the aerospace testing community to predict balance loads from strain-gage outputs during a wind tunnel test. The new method implicitly applies temperature corrections to the gage outputs during the load iteration process. Therefore, it can use uncorrected gage outputs directly as input for the load calculations. The new method is applied in several steps. First, balance calibration data is analyzed in the usual manner assuming that the balance temperature was kept constant during the calibration. Then, the temperature difference relative to the calibration temperature is introduced as a new independent variable for each strain--gage output. Therefore, sensors must exist near the strain--gages so that the required temperature differences can be measured during the wind tunnel test. In addition, the format of the regression coefficient matrix needs to be extended so that it can support the new independent variables. In the next step, the extended regression coefficient matrix of the original calibration data is modified by using the manufacturer specified temperature sensitivity of each strain--gage as the regression coefficient of the corresponding temperature difference variable. Finally, the modified regression coefficient matrix is converted to a data reduction matrix that the iterative analysis technique needs for the calculation of balance loads. Original calibration data and modified check load data of NASA's MC60D balance are used to illustrate the new method.

  15. Strain-Driven Nanoscale Phase Competition near the Antipolar-Nonpolar Phase Boundary in Bi0.7La0.3FeO3 Thin Films.

    PubMed

    Dedon, Liv R; Chen, Zuhuang; Gao, Ran; Qi, Yajun; Arenholz, Elke; Martin, Lane W

    2018-05-02

    Complex-oxide materials tuned to be near phase boundaries via chemistry/composition, temperature, pressure, etc. are known to exhibit large susceptibilities. Here, we observe a strain-driven nanoscale phase competition in epitaxially constrained Bi 0.7 La 0.3 FeO 3 thin films near the antipolar-nonpolar phase boundary and explore the evolution of the structural, dielectric, (anti)ferroelectric, and magnetic properties with strain. We find that compressive and tensile strains can stabilize an antipolar PbZrO 3 -like Pbam phase and a nonpolar Pnma orthorhombic phase, respectively. Heterostructures grown with little to no strain exhibit a self-assembled nanoscale mixture of the two orthorhombic phases, wherein the relative fraction of each phase can be modified with film thickness. Subsequent investigation of the dielectric and (anti)ferroelectric properties reveals an electric-field-driven phase transformation from the nonpolar phase to the antipolar phase. X-ray linear dichroism reveals that the antiferromagnetic-spin axes can be effectively modified by the strain-induced phase transition. This evolution of antiferromagnetic-spin axes can be leveraged in exchange coupling between the antiferromagnetic Bi 0.7 La 0.3 FeO 3 and a ferromagnetic Co 0.9 Fe 0.1 layer to tune the ferromagnetic easy axis of the Co 0.9 Fe 0.1 . These results demonstrate that besides chemical alloying, epitaxial strain is an alternative and effective way to modify subtle phase relations and tune physical properties in rare earth-alloyed BiFeO 3 . Furthermore, the observation of antiferroelectric-antiferromagnetic properties in the Pbam Bi 0.7 La 0.3 FeO 3 phase could be of significant scientific interest and great potential in magnetoelectric devices because of its dual antiferroic nature.

  16. Heterologous Protein Secretion in Lactobacilli with Modified pSIP Vectors

    PubMed Central

    Karlskås, Ingrid Lea; Maudal, Kristina; Axelsson, Lars; Rud, Ida; Eijsink, Vincent G. H.; Mathiesen, Geir

    2014-01-01

    We describe new variants of the modular pSIP-vectors for inducible gene expression and protein secretion in lactobacilli. The basic functionality of the pSIP system was tested in Lactobacillus strains representing 14 species using pSIP411, which harbors the broad-host-range Lactococcus lactis SH71rep replicon and a β-glucuronidase encoding reporter gene. In 10 species, the inducible gene expression system was functional. Based on these results, three pSIP vectors with different signal peptides were modified by replacing their narrow-host-range L. plantarum 256rep replicon with SH71rep and transformed into strains of five different species of Lactobacillus. All recombinant strains secreted the target protein NucA, albeit with varying production levels and secretion efficiencies. The Lp_3050 derived signal peptide generally resulted in the highest levels of secreted NucA. These modified pSIP vectors are useful tools for engineering a wide variety of Lactobacillus species. PMID:24614815

  17. Job strain, rank, and mental health in the UK Armed Forces.

    PubMed

    Fear, Nicola Townsend; Rubin, G James; Hatch, Stephani; Hull, Lisa; Jones, Margaret; Hotopf, Matthew; Wessely, Simon; Rona, Roberto J

    2009-01-01

    We assessed whether job demand and job control have independent effects on psychological symptoms or whether job control modifies effect of job demand; we also assessed whether rank modified associations between job strain and psychological symptoms. We used the Post Traumatic Stress Disorder (PTSD) Checklist (PCL-C), General Health Questionnaire-12 (GHQ-12), Chalder Fatigue Scale, a checklist of 53 physical symptoms, and the WHO's Alcohol Use Disorders Identification Test (AUDIT). Job control, job demand, and rank were independently associated with PTSD, common mental disorders, multiple physical symptoms, and fatigue, but not with severe alcohol problems. Job control and demand had additive effects on psychological symptoms. Commissioned officers had lower risk of caseness for psychological symptoms than other ranks. Adjustment for rank had negligible effect on level of association between job strain and psychological symptoms. Reported job strain and rank contributed independently to psychological symptoms.

  18. A Viscoelastic Hybrid Shell Finite Element

    NASA Technical Reports Server (NTRS)

    Johnson, Arthur

    1999-01-01

    An elastic large displacement thick-shell hybrid finite element is modified to allow for the calculation of viscoelastic stresses. Internal strain variables are introduced at he element's stress nodes and are employed to construct a viscous material model. First order ordinary differential equations relate the internal strain variables to the corresponding elastic strains at the stress nodes. The viscous stresses are computed from the internal strain variables using viscous moduli which are a fraction of the elastic moduli. The energy dissipated by the action of the viscous stresses in included in the mixed variational functional. Nonlinear quasi-static viscous equilibrium equations are then obtained. Previously developed Taylor expansions of the equilibrium equations are modified to include the viscous terms. A predictor-corrector time marching solution algorithm is employed to solve the algebraic-differential equations. The viscous shell element is employed to numerically simulate a stair-step loading and unloading of an aircraft tire in contact with a frictionless surface.

  19. Switching the mode of sucrose utilization by Saccharomyces cerevisiae

    PubMed Central

    Badotti, Fernanda; Dário, Marcelo G; Alves, Sergio L; Cordioli, Maria Luiza A; Miletti, Luiz C; de Araujo, Pedro S; Stambuk, Boris U

    2008-01-01

    Background Overflow metabolism is an undesirable characteristic of aerobic cultures of Saccharomyces cerevisiae during biomass-directed processes. It results from elevated sugar consumption rates that cause a high substrate conversion to ethanol and other bi-products, severely affecting cell physiology, bioprocess performance, and biomass yields. Fed-batch culture, where sucrose consumption rates are controlled by the external addition of sugar aiming at its low concentrations in the fermentor, is the classical bioprocessing alternative to prevent sugar fermentation by yeasts. However, fed-batch fermentations present drawbacks that could be overcome by simpler batch cultures at relatively high (e.g. 20 g/L) initial sugar concentrations. In this study, a S. cerevisiae strain lacking invertase activity was engineered to transport sucrose into the cells through a low-affinity and low-capacity sucrose-H+ symport activity, and the growth kinetics and biomass yields on sucrose analyzed using simple batch cultures. Results We have deleted from the genome of a S. cerevisiae strain lacking invertase the high-affinity sucrose-H+ symporter encoded by the AGT1 gene. This strain could still grow efficiently on sucrose due to a low-affinity and low-capacity sucrose-H+ symport activity mediated by the MALx1 maltose permeases, and its further intracellular hydrolysis by cytoplasmic maltases. Although sucrose consumption by this engineered yeast strain was slower than with the parental yeast strain, the cells grew efficiently on sucrose due to an increased respiration of the carbon source. Consequently, this engineered yeast strain produced less ethanol and 1.5 to 2 times more biomass when cultivated in simple batch mode using 20 g/L sucrose as the carbon source. Conclusion Higher cell densities during batch cultures on 20 g/L sucrose were achieved by using a S. cerevisiae strain engineered in the sucrose uptake system. Such result was accomplished by effectively reducing sucrose uptake by the yeast cells, avoiding overflow metabolism, with the concomitant reduction in ethanol production. The use of this modified yeast strain in simpler batch culture mode can be a viable option to more complicated traditional sucrose-limited fed-batch cultures for biomass-directed processes of S. cerevisiae. PMID:18304329

  20. Formate supplementation enhances folate-dependent nucleotide biosynthesis and prevents spina bifida in a mouse model of folic acid-resistant neural tube defects.

    PubMed

    Sudiwala, Sonia; De Castro, Sandra C P; Leung, Kit-Yi; Brosnan, John T; Brosnan, Margaret E; Mills, Kevin; Copp, Andrew J; Greene, Nicholas D E

    2016-07-01

    The curly tail mouse provides a model for neural tube defects (spina bifida and exencephaly) that are resistant to prevention by folic acid. The major ct gene, responsible for spina bifida, corresponds to a hypomorphic allele of grainyhead-like 3 (Grhl3) but the frequency of NTDs is strongly influenced by modifiers in the genetic background. Moreover, exencephaly in the curly tail strain is not prevented by reinstatement of Grhl3 expression. In the current study we found that expression of Mthfd1L, encoding a key component of mitochondrial folate one-carbon metabolism (FOCM), is significantly reduced in ct/ct embryos compared to a partially congenic wild-type strain. This expression change is not attributable to regulation by Grhl3 or the genetic background at the Mthfd1L locus. Mitochondrial FOCM provides one-carbon units as formate for FOCM reactions in the cytosol. We found that maternal supplementation with formate prevented NTDs in curly tail embryos and also resulted in increased litter size. Analysis of the folate profile of neurulation-stage embryos showed that formate supplementation resulted in an increased proportion of formyl-THF and THF but a reduction in proportion of 5-methyl THF. In contrast, THF decreased and 5-methyl THF was relatively more abundant in the liver of supplemented dams than in controls. In embryos cultured through the period of spinal neurulation, incorporation of labelled thymidine and adenine into genomic DNA was suppressed by supplemental formate, suggesting that de novo folate-dependent biosynthesis of nucleotides (thymidylate and purines) was enhanced. We hypothesise that reduced Mthfd1L expression may contribute to susceptibility to NTDs in the curly tail strain and that formate acts as a one-carbon donor to prevent NTDs. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  1. Classical Swine Fever Outbreak after Modified Live LOM Strain Vaccination in Naive Pigs, South Korea

    PubMed Central

    Je, Sang H.; Kwon, Taeyong; Yoo, Sung J.; Lee, Dong-Uk; Lee, SeungYoon; Richt, Juergen A.

    2018-01-01

    We report classical swine fever outbreaks occurring in naive pig herds on Jeju Island, South Korea, after the introduction of the LOM vaccine strain. Two isolates from sick pigs had >99% identity with the vaccine stain. LOM strain does not appear safe; its use in the vaccine should be reconsidered. PMID:29553332

  2. Fermentation of oat and soybean hull hydrolysates into ethanol and xylitol by recombinant industrial strains of Saccharomyces cerevisiae under diverse oxygen environments

    USDA-ARS?s Scientific Manuscript database

    In this study, we evaluated the capacity of recombinant industrial Saccharomyces cerevisiae YRH 396 and YRH 400 strains to ferment sugars from oat hull and soybean hull hydrolysates into ethanol and xylitol. The strains were genetically modified by chromosomal integration of Pichia stipitis XYLI/XYL...

  3. Pseudomonas aeruginosa-producing Metallo-β-lactamases (VIM, IMP, SME, and AIM) in the Clinical Isolates of Intensive Care Units, a University Hospital in Isfahan, Iran

    PubMed Central

    Khorvash, Farzin; Yazdani, Mohammadreza; Shabani, Shiva; Soudi, Aliasghar

    2017-01-01

    Background: Pseudomonas aeruginosa is a severe challenge for antimicrobial therapy, due to the chromosomal mutations or exhibition of intrinsic resistance to various antimicrobial agents such as most β-lactams. We undertook this study to evaluate the existence of SME, IMP, AIM, and VIM metallo-β-lactamases (MBL) encoding genes among P. aeruginosa strains isolated from Intensive Care Unit (ICU) patients in Al-Zahra Hospital in Isfahan, Iran. Materials and Methods: In a retrospective cross-sectional study that was conducted between March 2012 and April 2013, a total of 48 strains of P. aeruginosa were collected from clinical specimens of bedridden patients in ICU wards. Susceptibility test was performed by disc diffusion method. All of the meropenem-resistant strains were subjected to modified Hodge test for detection of carbapenemases. Multiplex polymerase chain reaction was performed for detection of blaVIM, blaIMP, blaAIM, and blaSME genes. Results: In disk diffusion method, imipenem and meropenem showed the most and colistin the least resistant antimicrobial agents against P. aeruginosa strains. Of the 48 isolates, 36 (75%) were multidrug resistant (MDR). Amplification of β-lactamase genes showed the presence of blaVIM genes in 7 (%14.6) strains and blaIMP genes in 15 (31.3%) strains. All of the isolates were negative for blaSME and blaAIM genes. We could not find any statistically significant difference among the presence of this gene and MDR positive, age, or source of the specimen. Conclusion: As patients with infections caused by MBL-producing bacteria are at an intensified risk of treatment failure, fast determination of these organisms is necessary. Our findings may provide useful insights in replace of the appropriate antibiotics and may also prevent MBLs mediated resistance problem. PMID:29285477

  4. SHuffle, a novel Escherichia coli protein expression strain capable of correctly folding disulfide bonded proteins in its cytoplasm

    PubMed Central

    2012-01-01

    Background Production of correctly disulfide bonded proteins to high yields remains a challenge. Recombinant protein expression in Escherichia coli is the popular choice, especially within the research community. While there is an ever growing demand for new expression strains, few strains are dedicated to post-translational modifications, such as disulfide bond formation. Thus, new protein expression strains must be engineered and the parameters involved in producing disulfide bonded proteins must be understood. Results We have engineered a new E. coli protein expression strain named SHuffle, dedicated to producing correctly disulfide bonded active proteins to high yields within its cytoplasm. This strain is based on the trxB gor suppressor strain SMG96 where its cytoplasmic reductive pathways have been diminished, allowing for the formation of disulfide bonds in the cytoplasm. We have further engineered a major improvement by integrating into its chromosome a signal sequenceless disulfide bond isomerase, DsbC. We probed the redox state of DsbC in the oxidizing cytoplasm and evaluated its role in assisting the formation of correctly folded multi-disulfide bonded proteins. We optimized protein expression conditions, varying temperature, induction conditions, strain background and the co-expression of various helper proteins. We found that temperature has the biggest impact on improving yields and that the E. coli B strain background of this strain was superior to the K12 version. We also discovered that auto-expression of substrate target proteins using this strain resulted in higher yields of active pure protein. Finally, we found that co-expression of mutant thioredoxins and PDI homologs improved yields of various substrate proteins. Conclusions This work is the first extensive characterization of the trxB gor suppressor strain. The results presented should help researchers design the appropriate protein expression conditions using SHuffle strains. PMID:22569138

  5. Fluorogenic Strain-Promoted Alkyne-Diazo Cycloadditions

    PubMed Central

    Friscourt, Frédéric; Fahrni, Christoph J.; Boons, Geert-Jan

    2016-01-01

    Fluorogenic reactions in which non- or weakly-fluorescent reagents produce highly fluorescent products are attractive for detecting a broad range of compounds in the fields of bio-conjugation and material sciences. We report here that Fl-DIBO, a dibenzocyclooctyne derivative modified with a cyclopropenone moiety, can undergo fast strain-promoted cycloadditions under catalyst-free conditions with azides, nitrones, nitrile oxides as well as mono- and disubstituted diazo-derivatives. While the reaction with nitrile oxides, nitrones and disubstituted diazo compounds gave cycloadducts with low quantum yield, monosubstituted diazo reagents produced 1H-pyrazole derivatives that exhibited a ~160-fold fluorescence enhancement over Fl-DIBO combined with a greater than 10,000-fold increase in brightness. Concluding from quantum chemical calculations, fluorescence quenching of 3H-pyrazoles, which are formed by reaction with disubstituted diazo-derivatives, is likely due to the presence of energetically low-lying (n,π*) states. The fluorogenic probe Fl-DIBO was successfully employed for the labeling of diazo-tagged proteins without detectable background signal. Diazo-derivatives are emerging as attractive reporters for the labeling of biomolecules and the studies presented here demonstrate that Fl-DIBO can be employed for visualizing such biomolecules without the need for probe washout. PMID:26330090

  6. Large-scale functional RNAi screen in C. elegans identifies genes that regulate the dysfunction of mutant polyglutamine neurons

    PubMed Central

    2012-01-01

    Background A central goal in Huntington's disease (HD) research is to identify and prioritize candidate targets for neuroprotective intervention, which requires genome-scale information on the modifiers of early-stage neuron injury in HD. Results Here, we performed a large-scale RNA interference screen in C. elegans strains that express N-terminal huntingtin (htt) in touch receptor neurons. These neurons control the response to light touch. Their function is strongly impaired by expanded polyglutamines (128Q) as shown by the nearly complete loss of touch response in adult animals, providing an in vivo model in which to manipulate the early phases of expanded-polyQ neurotoxicity. In total, 6034 genes were examined, revealing 662 gene inactivations that either reduce or aggravate defective touch response in 128Q animals. Several genes were previously implicated in HD or neurodegenerative disease, suggesting that this screen has effectively identified candidate targets for HD. Network-based analysis emphasized a subset of high-confidence modifier genes in pathways of interest in HD including metabolic, neurodevelopmental and pro-survival pathways. Finally, 49 modifiers of 128Q-neuron dysfunction that are dysregulated in the striatum of either R/2 or CHL2 HD mice, or both, were identified. Conclusions Collectively, these results highlight the relevance to HD pathogenesis, providing novel information on the potential therapeutic targets for neuroprotection in HD. PMID:22413862

  7. Is the association between high strain work and depressive symptoms modified by private life social support: a cohort study of 1,074 Danish employees?

    PubMed

    Madsen, Ida E H; Jorgensen, Anette F B; Borritz, Marianne; Nielsen, Martin L; Rugulies, Reiner

    2014-07-08

    Previous studies have shown that psychosocial working conditions characterized by high psychological demands and low decision latitude (i.e., high strain work) are associated with increased risk of depressive symptoms. Little is known, however, concerning how this association may be modified by factors outside the working environment. This article examines the modifying role of private life social support in the relation between high strain work and the development of severe depressive symptoms. Data were questionnaire-based, collected from a cross-occupational sample of 1,074 Danish employees. At baseline, all participants were free of severe depressive symptoms, measured by the Mental Health Inventory. High strain work was defined by the combination of high psychological demands at work and low control, measured with multi-dimensional scales. Private life social support was operationalized as the number of life domains with confidants and dichotomized as low (0-1 domains) or high (2 or more domains). Using logistic regression we examined the risk of onset of severe depressive symptoms, adjusting for sex, age, occupational position, and prior depressive symptoms. Separately, neither high strain work nor low private life social support statistically significantly predicted depressive symptoms. However, participants with joint exposure to high strain work and low private life social support had an Odds ratio (OR) for severe depressive symptoms of 3.41 (95% CI: 1.36-8.58), compared to participants with no work strain and high private life social support. There was no increased risk for participants with high strain work and high private life social support (OR = 1.32, 95% CI: 0.65-2.68). The interaction term for departure from additivity was, however, not statistically significant (p = 0.18). Our findings suggest that high strain work may increase risk of depressive symptoms in individuals with low private life social support, although the effect-modification was statistically non-significant. Larger studies are needed to further establish the role of private life social support in the relation between high strain work and depression.

  8. Job strain and binge eating among Brazilian workers participating in the ELSA-Brasil study: does BMI matter?

    PubMed Central

    Pena Gralle, Ana Paula Bruno; Barbosa Moreno, Arlinda; Lopes Juvanhol, Leidjaira; Mendes da Fonseca, Maria de Jesus; Prates Melo, Enirtes Caetano; Antunes Nunes, Maria Angélica; Toivanen, Susanna; Griep, Rosane Härter

    2017-01-01

    Objective: To assess the association between job strain and binge eating as well as the effect-modifying influence of body mass index (BMI) on this association. Methods: A total of 11,951 active civil servants from the multicenter Brazilian Longitudinal Study of Adult Health (ELSA-Brasil) was included in this cross-sectional analysis. Job strain was assessed using the Demand-Control-Support Questionnaire. Binge eating was defined as eating a large amount of food with a sense of lack of control over what and how much is eaten in less than 2 hours at least twice a week. Multiple logistic regression was used to determine the association between binge eating and job strain as well as its interaction with BMI. Results: After adjustment, and using low-strain job as the reference category, binge eating was associated with high-strain job (high demand/low control: odds ratio [OR]=1.58, 95% confidence interval [CI] 1.26-1.98), active job (high demand/high control: OR=1.35, 95% CI 1.07-1.70), and passive job (low demand/low control: OR=1.24, 95% CI 1.01-1.53). Psychological job demands were positively associated with binge eating (OR=1.04, 95% CI 1.01-1.07), while greater job control and social support at work were each inversely associated with binge eating (OR=0.95, 95% CI 0.92-0.97 and OR=0.96, 95% CI 0.94-0.98, respectively). BMI modified the association between job strain and binge eating: Heavier psychological job demands were associated with higher odds of binge eating among obese participants, while a stronger inverse association between job control and binge eating was seen among slimmer participants. Conclusions: Job strain increases the odds of binge eating and this association is modified by BMI. PMID:28163281

  9. Modulation of the norfloxacin resistance in Staphylococcus aureus by Cordia verbenaceae DC

    PubMed Central

    Matias, Edinardo F.F.; Santos, Karla K. A.; Falcão-Silva, Vivyanne S.; Siqueira-Júnior, José P.; Costa, José G. M.; Coutinho, Henrique D.M.

    2013-01-01

    Background & objectives: Several chemical compounds isolated from natural sources have antibacterial activity and some enhance the antibacterial activity of antibiotics reversing the natural resistance of bacteria to certain antibiotics. In this study, the hexane and methanol extract of Cordia verbenaceae were assessed for antibacterial activity alone and combinated with norfloxacin against the Staphylococcus aureus strain SA1199B. Methods: The minimum inhibitory concentration (MIC) of extracts was assayed using microdilution assay and the modulatory activity was evaluated using plate diffusion assay. Results: The MIC observed varied between 256 to >1024 μg/ml. However, the antibiotic activity of norfloxacin was enhanced in the presence of subinhibitory concentrations of hexane extract of C. verbenaceae (HECV). Interpretations & conclusions: Our results indicate that Cordia verbenaceae DC. can be a source of plant derived products with antibiotic modifying activity. PMID:23481069

  10. Strain-Specific Induction of Experimental Autoimmune Prostatitis (EAP) in Mice

    PubMed Central

    Jackson, Christopher M.; Flies, Dallas B.; Mosse, Claudio A.; Parwani, Anil; Hipkiss, Edward L.; Drake, Charles G.

    2013-01-01

    BACKGROUND Prostatitis, a clinical syndrome characterized by pelvic pain and inflammation, is common in adult males. Although several induced and spontaneous murine models of prostatitis have been explored, the role of genetic background on induction has not been well-defined. METHODS Using a standard methodology for the induction of experimental autoimmune prostatitis (EAP), we investigated both acute and chronic inflammation on several murine genetic backgrounds. RESULTS In our colony, nonobese diabetic (NOD) mice evinced spontaneous prostatitis that was not augmented by immunization with rat prostate extract (RPE). In contrast, the standard laboratory strain Balb/c developed chronic inflammation in response to RPE immunization. Development of EAP in other strains was variable. CONCLUSIONS These data suggest that Balb/c mice injected with RPE may provide a useful model for chronic prostatic inflammation. PMID:23129407

  11. FY17 Status Report on the Initial EPP Finite Element Analysis of Grade 91 Steel

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

    Messner, M. C.; Sham, T. -L.

    This report describes a modification to the elastic-perfectly plastic (EPP) strain limits design method to account for cyclic softening in Gr. 91 steel. The report demonstrates that the unmodified EPP strain limits method described in current ASME code case is not conservative for materials with substantial cyclic softening behavior like Gr. 91 steel. However, the EPP strain limits method can be modified to be conservative for softening materials by using softened isochronous stress-strain curves in place of the standard curves developed from unsoftened creep experiments. The report provides softened curves derived from inelastic material simulations and factors describing the transformationmore » of unsoftened curves to a softened state. Furthermore, the report outlines a method for deriving these factors directly from creep/fatigue tests. If the material softening saturates the proposed EPP strain limits method can be further simplified, providing a methodology based on temperature-dependent softening factors that could be implemented in an ASME code case allowing the use of the EPP strain limits method with Gr. 91. Finally, the report demonstrates the conservatism of the modified method when applied to inelastic simulation results and two bar experiments.« less

  12. Incorporation of Mean Stress Effects into the Micromechanical Analysis of the High Strain Rate Response of Polymer Matrix Composites

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.; Roberts, Gary D.; Gilat, Amos

    2002-01-01

    The results presented here are part of an ongoing research program, to develop strain rate dependent deformation and failure models for the analysis of polymer matrix composites subject to high strain rate impact loads. A micromechanics approach is employed in this work, in which state variable constitutive equations originally developed for metals have been modified to model the deformation of the polymer matrix, and a strength of materials based micromechanics method is used to predict the effective response of the composite. In the analysis of the inelastic deformation of the polymer matrix, the definitions of the effective stress and effective inelastic strain have been modified in order to account for the effect of hydrostatic stresses, which are significant in polymers. Two representative polymers, a toughened epoxy and a brittle epoxy, are characterized through the use of data from tensile and shear tests across a variety of strain rates. Results computed by using the developed constitutive equations correlate well with data generated via experiments. The procedure used to incorporate the constitutive equations within a micromechanics method is presented, and sample calculations of the deformation response of a composite for various fiber orientations and strain rates are discussed.

  13. TSCA Experimental Release Application Approved for Pseudomonas putida Strains (fact sheet)

    EPA Pesticide Factsheets

    In 1998, EPA approved the TERAs R98-0004/5 submitted by the National Explosives Waste Technology & Evaluation Center (NEWTEC) and the Oak Ridge National Laboratory for field trials of two modified strains of Pseudomonas putida (P.putida).

  14. Evolution of Dislocation Density During Tensile Deformation of BH220 Steel at Different Pre-strain Conditions

    NASA Astrophysics Data System (ADS)

    Seth, Prem Prakash; Das, A.; Bar, H. N.; Sivaprasad, S.; Basu, A.; Dutta, K.

    2015-07-01

    Tensile behavior of BH220 steel with different pre-strain conditions (2 and 8%) followed by bake hardening was studied at different strain rates (0.001 and 0.1/s). Dislocation densities of the deformed specimens were successfully estimated from x-ray diffraction profile analysis using the modified Williamson-Hall equation. The results indicate that other than 2% pre-strain the dislocation density increases with increase in pre-strain level as well as with strain rate. The decrease in the dislocation density in 2% pre-strain condition without any drop in strength value is attributed to the characteristic dislocation feature formed during pre-straining.

  15. Formation and Migration Energies of Interstitials in Silicon Under Strain Conditions

    NASA Technical Reports Server (NTRS)

    Halicioglu, Timur; Barnett, David M.

    1999-01-01

    Simulation calculations are conducted for Si substrates to analyze formation and diffusion energies of interstitials under strain condition using statics methods .based on a Stillinger-Weber type potential function. Defects in the vicinity of the surface region and in the bulk are examined, and the role played by compressive and tensile strains on the energetics of interstitials is investigated. Results indicate that strain alters defect energetics which, in turn, modifies their diffusion characteristics.

  16. Effects of polyaluminum chloride and lanthanum-modified bentonite on the growth rates of three Cylindrospermopsis raciborskii strains

    PubMed Central

    van Oosterhout, Frank; Becker, Vanessa; Attayde, José Luiz; Lürling, Miquel

    2018-01-01

    In tropical and subtropical lakes, eutrophication often leads to nuisance blooms of Cylindrospermopsis raciborskii. In laboratory experiments, we tested the combined effects of flocculant polyaluminum chloride (PAC) and lanthanum-modified bentonite (LMB) on the sinking and growth rates of three C. raciborskii strains. We tested the hypothesis that the combination of PAC and LMB would (1) effectively sink C. raciborskii in a test tube experiment and (2) impair C. raciborskii growth, irrespective of the biomass of the inoculum (bloom) and the strain in the growth experiment. We tested the recommended (LMB1) and a three-times higher dose of LMB (LMB3). The combined addition of PAC and LMB enhanced the sedimentation of all C. raciborskii strains. Moreover, both the PAC and LMB doses decreased the phosphate concentration. PAC and LMB1 decreased the growth rate of all strains, but the efficacy depended on the biomass and strain. The combined addition of PAC and LMB3 inhibited the growth of all strains independently of the biomass and strain. We conclude that a low dose of PAC in combination with the recommended dose of LMB decreases C. raciborskii blooms and that the efficiency of the technique depends on the biomass of the bloom. A higher dose of LMB is needed to obtain a more efficient control of C. raciborskii blooms. PMID:29614118

  17. Effects of polyaluminum chloride and lanthanum-modified bentonite on the growth rates of three Cylindrospermopsis raciborskii strains.

    PubMed

    Araújo, Fabiana; van Oosterhout, Frank; Becker, Vanessa; Attayde, José Luiz; Lürling, Miquel

    2018-01-01

    In tropical and subtropical lakes, eutrophication often leads to nuisance blooms of Cylindrospermopsis raciborskii. In laboratory experiments, we tested the combined effects of flocculant polyaluminum chloride (PAC) and lanthanum-modified bentonite (LMB) on the sinking and growth rates of three C. raciborskii strains. We tested the hypothesis that the combination of PAC and LMB would (1) effectively sink C. raciborskii in a test tube experiment and (2) impair C. raciborskii growth, irrespective of the biomass of the inoculum (bloom) and the strain in the growth experiment. We tested the recommended (LMB1) and a three-times higher dose of LMB (LMB3). The combined addition of PAC and LMB enhanced the sedimentation of all C. raciborskii strains. Moreover, both the PAC and LMB doses decreased the phosphate concentration. PAC and LMB1 decreased the growth rate of all strains, but the efficacy depended on the biomass and strain. The combined addition of PAC and LMB3 inhibited the growth of all strains independently of the biomass and strain. We conclude that a low dose of PAC in combination with the recommended dose of LMB decreases C. raciborskii blooms and that the efficiency of the technique depends on the biomass of the bloom. A higher dose of LMB is needed to obtain a more efficient control of C. raciborskii blooms.

  18. Modeling of a Surface Acoustic Wave Strain Sensor

    NASA Technical Reports Server (NTRS)

    Wilson, W. C.; Atkinson, Gary M.

    2010-01-01

    NASA Langley Research Center is investigating Surface Acoustic Wave (SAW) sensor technology for harsh environments aimed at aerospace applications. To aid in development of sensors a model of a SAW strain sensor has been developed. The new model extends the modified matrix method to include the response of Orthogonal Frequency Coded (OFC) reflectors and the response of SAW devices to strain. These results show that the model accurately captures the strain response of a SAW sensor on a Langasite substrate. The results of the model of a SAW Strain Sensor on Langasite are presented

  19. Reduction of Phytate in Soy Drink by Fermentation with Lactobacillus casei Expressing Phytases From Bifidobacteria.

    PubMed

    García-Mantrana, Izaskun; Monedero, Vicente; Haros, Monika

    2015-09-01

    Plant-based food products can be modified by fermentation to improve flavour and the concentration of some biologically active compounds, but also to increase the mineral availability by eliminating anti-nutrient substances such as phytates. The objective of this study was to develop a fermented soybean drink with improved nutritional quality and source of probiotic bacteria by including as starter for fermentation Lactobacillus casei strains modified to produce phytase enzymes from bifidobacteria. The L. casei strains showed a good adaptation to develop in the soy drink but they needed the addition of external carbohydrates to give rise to an efficient acidification. The strain expressing the Bifidobacterium pseudocatenulatum phytase was able to degrade more than 90 % phytate during product fermentation, whereas expression of Bifidobacterium longum spp. infantis phytase only led to 65 % hydrolysis. In both cases, accumulation of myo-inositol triphosphates was observed. In addition, the hydrolysis of phytate in soy drink fermented with the L. casei strain expressing the B. pseudocatenulatum phytase resulted in phytate/mineral ratios for Fe (0.35) and Zn (2.4), which were below the critical values for reduced mineral bioavailability in humans. This investigation showed the ability of modified L. casei to produce enzymes with technological relevance in the design of new functional foods.

  20. Modified Displacement Transfer Functions for Deformed Shape Predictions of Slender Curved Structures with Varying Curvatives

    NASA Technical Reports Server (NTRS)

    Ko, William L.; Fleischer, Van Tran

    2014-01-01

    To eliminate the need to use finite-element modeling for structure shape predictions, a new method was invented. This method is to use the Displacement Transfer Functions to transform the measured surface strains into deflections for mapping out overall structural deformed shapes. The Displacement Transfer Functions are expressed in terms of rectilinearly distributed surface strains, and contain no material properties. This report is to apply the patented method to the shape predictions of non-symmetrically loaded slender curved structures with different curvatures up to a full circle. Because the measured surface strains are not available, finite-element analysis had to be used to analytically generate the surface strains. Previously formulated straight-beam Displacement Transfer Functions were modified by introducing the curvature-effect correction terms. Through single-point or dual-point collocations with finite-elementgenerated deflection curves, functional forms of the curvature-effect correction terms were empirically established. The resulting modified Displacement Transfer Functions can then provide quite accurate shape predictions. Also, the uniform straight-beam Displacement Transfer Function was applied to the shape predictions of a section-cut of a generic capsule (GC) outer curved sandwich wall. The resulting GC shape predictions are quite accurate in partial regions where the radius of curvature does not change sharply.

  1. Shear-strain energy rate distribution caused by the interplate locking along the Nankai Trough, southwest Japan: An integration analysis using stress tensor inversion and slip deficit inversion

    NASA Astrophysics Data System (ADS)

    Saito, T.; Noda, A.; Yoshida, K.; Tanaka, S.

    2017-12-01

    In the Nankai Trough, southwest Japan, the Philippine Sea Plate descends beneath the Eurasian plate. The locking, or the slip deficit, on the plate interface causes stress fluctuation in the inland area. The interplate locking does not always result in stress accumulation but also causes stress release. The stress increase/decrease is not determined only from the stress fluctuation but also depends on the background stress, in particular, its orientation. This study proposes a method to estimate the shear-strain energy increase/decrease distribution caused by the interplate locking. We at first investigated the background stress field in and around the Nankai Trough. The spatial distribution of the principal stress orientations and the stress ratio were estimated by analysis of 130,000 focal mechanisms of small earthquakes (e.g., Yoshida et al. 2015 Tectonophysics). For example, in an area called Chugoku region, the maximum and minimum compression axes were E-W and N-S directions, respectively. We also estimated the slip-deficit rate at the plate interface by analyzing GNSS data and calculated the stress fluctuation due to the deficit (e.g., Noda et al. 2013 GJI). The interplate locking causes the maximum compression in the direction of plate convergence. This is significantly different from the orientations of the background stress characterized by the E-W compressional strike-slip stress regime.. By combining the results of the background stress and the stress fluctuation, we made a map indicating the shear-strain energy change due to the interplate locking. In the Chugoku region, the shear-strain energy decreases due to the interplate locking. This is because the N-S compressional stress caused by the interplate locking compensates the N-S extensional stress in the background. The shear-strain energy increases in some parts of the analyzed areas. By statistically comparing the shear strain energy rate with the seismicity in the inland area, we found that the seismicity tends to be high where the interplate locking increases the shear-strain energy. Our results suggest that the stress fluctuation due to the interplate locking is not dominant in the background stress but surely contributes to the inland seismicity in southwest Japan.

  2. Demonstration test of burner liner strain measuring system

    NASA Technical Reports Server (NTRS)

    Stetson, K. A.

    1984-01-01

    A demonstration test was conducted for two systems of static strain measurement that had been shown to have potential for application jet engine combustors. A modified JT12D combustor was operated in a jet burner test stand while subjected simultaneously to both systems of instrumentation, i.e., Kanthal A-1 wire strain gages and laser speckle photography. A section of the burner was removed for installation and calibration of the wire gages, and welded back into the burner. The burner test rig was modified to provide a viewing port for the laser speckle photography such that the instrumented section could be observed during operation. Six out of ten wire gages survived testing and showed excellent repeatability. The extensive precalibration procedures were shown to be effective in compensating for the large apparent strains associated with these gages. Although all portions of the speckle photography system operated satisfactorily, a problem was encountered in the form of optical inhomogeneities in the hot, high-pressure gas flowing by the combustor case which generate large and random apparent strain distributions.

  3. The role of genetic background in susceptibility to chemical warfare nerve agents across rodent and non-human primate models.

    PubMed

    Matson, Liana M; McCarren, Hilary S; Cadieux, C Linn; Cerasoli, Douglas M; McDonough, John H

    2018-01-15

    Genetics likely play a role in various responses to nerve agent exposure, as genetic background plays an important role in behavioral, neurological, and physiological responses to environmental stimuli. Mouse strains or selected lines can be used to identify susceptibility based on background genetic features to nerve agent exposure. Additional genetic techniques can then be used to identify mechanisms underlying resistance and sensitivity, with the ultimate goal of developing more effective and targeted therapies. Here, we discuss the available literature on strain and selected line differences in cholinesterase activity levels and response to nerve agent-induced toxicity and seizures. We also discuss the available cholinesterase and toxicity literature across different non-human primate species. The available data suggest that robust genetic differences exist in cholinesterase activity, nerve agent-induced toxicity, and chemical-induced seizures. Available cholinesterase data suggest that acetylcholinesterase activity differs across strains, but are limited by the paucity of carboxylesterase data in strains and selected lines. Toxicity and seizures, two outcomes of nerve agent exposure, have not been fully evaluated for genetic differences, and thus further studies are required to understand baseline strain and selected line differences. Published by Elsevier B.V.

  4. Phase shift of oscillatory magnetoresistance in a double-cross thin film structure of La0.3Pr0.4Ca0.3MnO3 via strain-engineered elongation of electronic domains

    NASA Astrophysics Data System (ADS)

    Alagoz, H. S.; Prasad, B.; Jeon, J.; Blamire, M. G.; Chow, K. H.; Jung, J.

    2018-02-01

    The subtle balance between the competing electronic phases in manganites due to complex interplay between spin, charge, and orbital degrees of freedom could allow one to modify the properties of electronically phase separated systems. In this paper, we show that the phase shift in the oscillatory magnetoresistance ρ (θ ) can be modified by engineering strain driven elongation of electronic domains in La0.3Pr0.4Ca0.3MnO3 (LPCMO) thin films. Strain-driven elongation of magnetic domains can produce different percolation paths and hence different anisotropic magnetoresistance responses. This tunability provides a unique control that is unattainable in conventional 3 d ferromagnetic metals and alloys.

  5. An Ahemolytic Pneumolysin of Streptococcus Pneumoniae Manipulates Human Innate and CD4+ T-Cell Responses and Reduces Resistance to Colonization in Mice in a Serotype-Independent Manner

    PubMed Central

    Khan, M. Nadeem; Coleman, John Robert; Vernatter, Joshua; Varshney, Avanish Kumar; Dufaud, Chad; Pirofski, Liise-anne

    2014-01-01

    Background. Some Streptococcus pneumoniae serotypes express an ahemolytic pneumolysin (PLYa). Serotypes that commonly express PLYa, including serotype 8 (ST8) and ST1, are often associated with a low prevalence during colonization but a higher propensity to cause invasive disease. We sought to study the host response to ST8 PLYa in a homologous and heterologous capsular background. Methods. We genetically exchanged the PLYa of ST8 strain 6308 with the hemolytic PLY (PLYh) of ST3 A66.1 and vice versa and determined the impact of the exchange on nasopharyngeal colonization in mice. Then, to compare the response of human cells to PLYa-expressing and PLYh-expressing strains, we infected human peripheral blood mononuclear cells (PBMCs) with PLY-switched strains and assessed dendritic cell and CD4+ T-cell responses by intracellular cytokine staining. Result. Mice colonized with PLYa-expressing strains had significantly higher colonization densities than those colonized with PLYh-expressing strains, irrespective of capsular background. Compared with infection of PBMCs with PLYh-expressing strains, infection with PLYa-expressing strains induced diminished innate (dendritic cell cytokines, costimulatory receptor, and apoptotic) and adaptive (CD4+ T-cell proliferative and memory interleukin 17A) responses. Conclusion. Our findings demonstrate that PLYa has the potential to manipulate host immunity irrespective of capsule type. PLY exchange between STs expressing PLYa and PLYh could lead to unexpected colonization or invasion phenotypes. PMID:25001458

  6. Genetic dissection of quantitative trait locus for ethanol sensitivity in long- and short-sleep mice.

    PubMed

    Bennett, B; Carosone-Link, P; Beeson, M; Gordon, L; Phares-Zook, N; Johnson, T E

    2008-08-01

    Interval-specific congenic strains (ISCS) allow fine mapping of a quantitative trait locus (QTL), narrowing its confidence interval by an order of magnitude or more. In earlier work, we mapped four QTL specifying differential ethanol sensitivity, assessed by loss of righting reflex because of ethanol (LORE), in the inbred long-sleep (ILS) and inbred short-sleep (ISS) strains, accounting for approximately 50% of the genetic variance for this trait. Subsequently, we generated reciprocal congenic strains in which each full QTL interval from ILS was bred onto the ISS background and vice versa. An earlier paper reported construction and results of the ISCS on the ISS background; here, we describe this process and report results on the ILS background. We developed multiple ISCS for each Lore QTL in which the QTL interval was broken into a number of smaller intervals. For each of the four QTL regions (chromosomes 1, 2, 11 and 15), we were successful in reducing the intervals significantly. Multiple, positive strains were overlapped to generate a single, reduced interval. Subsequently, this reduced region was overlaid on previous reductions from the ISS background congenics, resulting in substantial reductions in all QTL regions by approximately 75% from the initial mapping study. Genes with sequence or expression polymorphisms in the reduced intervals are potential candidates; evidence for these is presented. Genetic background effects can be important in detection of single QTL; combining this information with the generation of congenics on both backgrounds, as described here, is a powerful approach for fine mapping QTL.

  7. Technological characterization and survival of the exopolysaccharide-producing strain Lactobacillus delbrueckii subsp. lactis 193 and its bile-resistant derivative 193+ in simulated gastric and intestinal juices.

    PubMed

    Burns, Patricia; Vinderola, Gabriel; Reinheimer, Jorge; Cuesta, Isabel; de Los Reyes-Gavilán, Clara G; Ruas-Madiedo, Patricia

    2011-08-01

    The capacity of lactic acid bacteria to produce exopolysaccharides (EPS) conferring microorganisms a ropy phenotype could be an interesting feature from a technological point of view. Progressive adaptation to bile salts might render some lactobacilli able to overcome physiological gut barriers but could also modify functional properties of the strain, including the production of EPS. In this work some technological properties and the survival ability in simulated gastrointestinal conditions of Lactobacillus delbrueckii subsp. lactis 193, and Lb. delbrueckii subsp. lactis 193+, a strain with stable bile-resistant phenotype derived thereof, were characterized in milk in order to know whether the acquisition of resistance to bile could modify some characteristics of the microorganism. Both strains were able to grow and acidify milk similarly; however the production of ethanol increased at the expense of the aroma compound acetaldehyde in milk fermented by the strain 193+, with respect to milk fermented by the strain 193. Both microorganisms produced a heteropolysaccharide composed of glucose and galactose, and were able to increase the viscosity of fermented milks. In spite of the higher production yield of EPS by the bile-resistant strain 193+, it displayed a lower ability to increase viscosity than Lb. delbrueckii subsp. lactis 193. Milk increased survival in simulated gastric juice; the presence of bile improved adhesion to the intestinal cell line HT29-MTX in both strains. However, the acquisition of a stable resistance phenotype did not improve survival in simulated gastric and intestinal conditions or the adhesion to the intestinal cell line HT29-MTX. Thus, Lb. delbrueckii subsp. lactis 193 presents suitable technological properties for the manufacture of fermented dairy products; the acquisition of a stable bile-resistant phenotype modified some properties of the microorganism. This suggests that the possible use of bile-resistant derivative strains should be carefully evaluated in each specific application considering the influence that the acquisition of a stable bile-resistant phenotype could have in survival ability in gastric and intestinal conditions and in technological properties.

  8. Altered trophoblast proliferation is insufficient to account for placental dysfunction in Egfr null embryos

    PubMed Central

    Dackor, J.; Strunk, K. E.; Wehmeyer, M. M.; Threadgill, D. W.

    2007-01-01

    Homozygosity for the Egfrtm1Mag null allele in mice leads to genetic background dependent placental abnormalities and embryonic lethality. Molecular mechanisms or genetic modifiers that differentiate strains with surviving versus non-surviving Egfr nullizygous embryos have yet to be identified. Egfr transcripts in wildtype placenta was quantified by ribonuclease protection assay (RPA) and the lowest level of Egfr mRNA expression was found to coincide with Egfrtm1Mag homozygous lethality. Immunohistochemical analysis of ERBB family receptors, ERBB2, ERBB3, and ERBB4, showed similar expression between Egfr wildtype and null placentas indicating that Egfr null trophoblast do not up-regulate these receptors to compensate for EGFR deficiency. Significantly fewer numbers of bromodeoxyuridine (BrdU) positive trophoblast were observed in Egfr nullizygous placentas and Cdc25a and Myc, genes associated with proliferation, were significantly down-regulated in null placentas. However, strains with both mild and severe placental phenotypes exhibit reduced proliferation suggesting that this defect alone does not account for strain-specific embryonic lethality. Consistent with this hypothesis, intercrosses generating mice null for cell cycle checkpoint genes (Trp53, Rb1, Cdkn1a, Cdkn1b or Cdkn2c) in combination with Egfr deficiency did not increase survival of Egfr nullizygous embryos. Since complete development of the spongiotrophoblast compartment is not required for survival of Egfr nullizygous embryos, reduction of this layer that is commonly observed in Egfr nullizygous placentas likely accounts for the decrease in proliferation. PMID:17822758

  9. Analytical Modeling of the High Strain Rate Deformation of Polymer Matrix Composites

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.; Roberts, Gary D.; Gilat, Amos

    2003-01-01

    The results presented here are part of an ongoing research program to develop strain rate dependent deformation and failure models for the analysis of polymer matrix composites subject to high strain rate impact loads. State variable constitutive equations originally developed for metals have been modified in order to model the nonlinear, strain rate dependent deformation of polymeric matrix materials. To account for the effects of hydrostatic stresses, which are significant in polymers, the classical 5 plasticity theory definitions of effective stress and effective plastic strain are modified by applying variations of the Drucker-Prager yield criterion. To verify the revised formulation, the shear and tensile deformation of a representative toughened epoxy is analyzed across a wide range of strain rates (from quasi-static to high strain rates) and the results are compared to experimentally obtained values. For the analyzed polymers, both the tensile and shear stress-strain curves computed using the analytical model correlate well with values obtained through experimental tests. The polymer constitutive equations are implemented within a strength of materials based micromechanics method to predict the nonlinear, strain rate dependent deformation of polymer matrix composites. In the micromechanics, the unit cell is divided up into a number of independently analyzed slices, and laminate theory is then applied to obtain the effective deformation of the unit cell. The composite mechanics are verified by analyzing the deformation of a representative polymer matrix composite (composed using the representative polymer analyzed for the correlation of the polymer constitutive equations) for several fiber orientation angles across a variety of strain rates. The computed values compare favorably to experimentally obtained results.

  10. Stem Cell Therapy with Overexpressed VEGF and PDGF Genes Improves Cardiac Function in a Rat Infarct Model

    PubMed Central

    Das, Hiranmoy; George, Jon C.; Joseph, Matthew; Das, Manjusri; Abdulhameed, Nasreen; Blitz, Anna; Khan, Mahmood; Sakthivel, Ramasamy; Mao, Hai-Quan; Hoit, Brian D.; Kuppusamy, Periannan; Pompili, Vincent J.

    2009-01-01

    Background Therapeutic potential was evaluated in a rat model of myocardial infarction using nanofiber-expanded human cord blood derived hematopoietic stem cells (CD133+/CD34+) genetically modified with VEGF plus PDGF genes (VIP). Methods and Findings Myocardial function was monitored every two weeks up to six weeks after therapy. Echocardiography revealed time dependent improvement of left ventricular function evaluated by M-mode, fractional shortening, anterior wall tissue velocity, wall motion score index, strain and strain rate in animals treated with VEGF plus PDGF overexpressed stem cells (VIP) compared to nanofiber expanded cells (Exp), freshly isolated cells (FCB) or media control (Media). Improvement observed was as follows: VIP>Exp> FCB>media. Similar trend was noticed in the exercise capacity of rats on a treadmill. These findings correlated with significantly increased neovascularization in ischemic tissue and markedly reduced infarct area in animals in the VIP group. Stem cells in addition to their usual homing sites such as lung, spleen, bone marrow and liver, also migrated to sites of myocardial ischemia. The improvement of cardiac function correlated with expression of heart tissue connexin 43, a gap junctional protein, and heart tissue angiogenesis related protein molecules like VEGF, pNOS3, NOS2 and GSK3. There was no evidence of upregulation in the molecules of oncogenic potential in genetically modified or other stem cell therapy groups. Conclusion Regenerative therapy using nanofiber-expanded hematopoietic stem cells with overexpression of VEGF and PDGF has a favorable impact on the improvement of rat myocardial function accompanied by upregulation of tissue connexin 43 and pro-angiogenic molecules after infarction. PMID:19809493

  11. Modeling the Nonlinear, Strain Rate Dependent Deformation of Shuttle Leading Edge Materials with Hydrostatic Stress Effects Included

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.; Carney, Kelly S.

    2004-01-01

    An analysis method based on a deformation (as opposed to damage) approach has been developed to model the strain rate dependent, nonlinear deformation of woven ceramic matrix composites, such as the Reinforced Carbon Carbon (RCC) material used on the leading edges of the Space Shuttle. In the developed model, the differences in the tension and compression deformation behaviors have also been accounted for. State variable viscoplastic equations originally developed for metals have been modified to analyze the ceramic matrix composites. To account for the tension/compression asymmetry in the material, the effective stress and effective inelastic strain definitions have been modified. The equations have also been modified to account for the fact that in an orthotropic composite the in-plane shear response is independent of the stiffness in the normal directions. The developed equations have been implemented into LS-DYNA through the use of user defined subroutines (UMATs). Several sample qualitative calculations have been conducted, which demonstrate the ability of the model to qualitatively capture the features of the deformation response present in woven ceramic matrix composites.

  12. Strain commonalities and differences in response-outcome decision making in mice

    PubMed Central

    Zimmermann, Kelsey S.; Hsu, Chia-Chun; Gourley, Shannon L.

    2016-01-01

    The ability to select between actions that are more vs. less likely to be reinforced is necessary for survival and navigation of a changing environment. A task termed “response-outcome contingency degradation” can be used in the laboratory to determine whether rodents behave according to such goal-directed response strategies. In one iteration of this task, rodents are trained to perform two food-reinforced behaviors, then the predictive relationship between one instrumental response and the associated outcome is modified by providing the reinforcer associated with that response non-contingently. During a subsequent probe test, animals can select between the two trained responses. Preferential engagement of the behavior most likely to be reinforced is considered goal-directed, while non-selective responding is considered a failure in response-outcome conditioning, or “habitual.” This test has largely been used with rats, and less so with mice. Here we compiled data collected from several cohorts of mice tested in our lab between 2012-2015. Mice were bred on either a C57BL/6 or predominantly BALB/c strain background. We report that both strains of mice can use information acquired as a result of instrumental contingency degradation training to select amongst multiple response options the response most likely to be reinforced. Mice differ, however, during the training sessions when the familiar response-outcome contingency is being violated. BALB/c mice readily generate perseverative or habit-like response strategies when the only available response is unlikely to be reinforced, while C57BL/6 mice more readily inhibit responding. These findings provide evidence of strain differences in response strategies when an anticipated reinforcer is unlikely to be delivered. PMID:27003118

  13. Progress of a Cross-correlation Based Optical Strain Measurement Technique for Detecting Radial Growth on a Rotating Disk

    NASA Technical Reports Server (NTRS)

    Clem, Michelle M.; Woike, Mark; Abdul-Aziz, Ali

    2013-01-01

    The Aeronautical Sciences Project under NASAs Fundamental Aeronautics Program is extremely interested in the development of fault detection technologies, such as optical surface measurements in the internal parts of a flow path, for in situ health monitoring of gas turbine engines. In situ health monitoring has the potential to detect flaws, i.e. cracks in key components, such as engine turbine disks, before the flaws lead to catastrophic failure. In the present study, a cross-correlation imaging technique is investigated in a proof-of-concept study as a possible optical technique to measure the radial growth and strain field on an already cracked sub-scale turbine engine disk under loaded conditions in the NASA Glenn Research Centers High Precision Rotordynamics Laboratory. The optical strain measurement technique under investigation offers potential fault detection using an applied background consisting of a high-contrast random speckle pattern and imaging the background under unloaded and loaded conditions with a CCD camera. Spinning the cracked disk at high speeds induces an external load, resulting in a radial growth of the disk of approximately 50.8-m in the flawed region and hence, a localized strain field. When imaging the cracked disk under static conditions, the disk will appear shifted. The resulting background displacements between the two images will then be measured using the two-dimensional cross-correlation algorithms implemented in standard Particle Image Velocimetry (PIV) software to track the disk growth, which facilitates calculation of the localized strain field. In order to develop and validate this optical strain measurement technique an initial proof-of-concept experiment is carried out in a controlled environment. Using PIV optimization principles and guidelines, three potential backgrounds, for future use on the rotating disk, are developed and investigated in the controlled experiment. A range of known shifts are induced on the backgrounds; reference and data images are acquired before and after the induced shift, respectively, and the images are processed using the cross- correlation algorithms in order to determine the background displacements. The effectiveness of each background at resolving the known shift is evaluated and discussed in order to choose to the most suitable background to be implemented onto a rotating disk in the Rotordynamics Lab. Although testing on the rotating disk has not yet been performed, the driving principles behind the development of the present optical technique are based upon critical aspects of the future experiment, such as the amount of expected radial growth, disk analysis, and experimental design and are therefore addressed in the paper.

  14. Dynamic Deformation Behavior of Soft Material Using Shpb Technique and Pulse Shaper

    NASA Astrophysics Data System (ADS)

    Lee, Ouk Sub; Cho, Kyu Sang; Kim, Sung Hyun; Han, Yong Hwan

    This paper presents a modified Split Hopkinson Pressure Bar (SHPB) technique to obtain compressive stress strain data for NBR rubber materials. An experimental technique with a modified the conventional SHPB has been developed for measuring the compressive stress strain responses of materials with low mechanical impedance and low compressive strengths, such as the rubber and the polymeric material. This paper uses an aluminum pressure bar to achieve a closer impedance match between the pressure bar and the specimen materials. In addition, a pulse shaper is utilized to lengthen the rising time of the incident pulse to ensure dynamic stress equilibrium and homogeneous deformation of NBR rubber materials. It is found that the modified technique can determine the dynamic deformation behavior of rubbers more accurately.

  15. Preparation for emergence of an Eastern European porcine reproductive and respiratory syndrome virus (PRRSV) strain in Western Europe: Immunization with modified live virus vaccines or a field strain confers partial protection.

    PubMed

    Renson, P; Fablet, C; Le Dimna, M; Mahé, S; Touzain, F; Blanchard, Y; Paboeuf, F; Rose, N; Bourry, O

    2017-05-01

    The porcine reproductive and respiratory syndrome virus (PRRSV) causes huge economic losses for the swine industry worldwide. In the past several years, highly pathogenic strains that lead to even greater losses have emerged. For the Western European swine industry, one threat is the possible introduction of Eastern European PRRSV strains (example Lena genotype 1.3) which were shown to be more virulent than common Western resident strains under experimental conditions. To prepare for the possible emergence of this strain in Western Europe, we immunized piglets with a Western European PRRSV field strain (Finistere: Fini, genotype 1.1), a new genotype 1 commercial modified live virus (MLV) vaccine (MLV1) or a genotype 2 commercial MLV vaccine (MLV2) to evaluate and compare the level of protection that these strains conferred upon challenge with the Lena strain 4 weeks later. Results show that immunization with Fini, MLV1 or MLV2 strains shortened the Lena-induced hyperthermia. In the Fini group, a positive effect was also demonstrated in growth performance. The level of Lena viremia was reduced for all immunized groups (significantly so for Fini and MLV2). This reduction in Lena viremia was correlated with the level of Lena-specific IFNγ-secreting cells. In conclusion, we showed that a commercial MLV vaccine of genotype 1 or 2, as well as a field strain of genotype 1.1 may provide partial clinical and virological protection upon challenge with the Lena strain. The cross-protection induced by these immunizing strains was not related with the level of genetic similarity to the Lena strain. The slightly higher level of protection established with the field strain is attributed to a better cell-mediated immune response. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Verification and application of the Iosipescu shear test method

    NASA Technical Reports Server (NTRS)

    Walrath, D. E.; Adams, D. F.

    1984-01-01

    Finite element models were used to study the effects of notch angle variations on the stress state within an Iosipescu shear test speciment. These analytical results were also studied to determine the feasibility of using strain gage rosettes and a modified extensometer to measure shear strains in this test specimen. Analytical results indicate that notch angle variations produced only small differences in simulated shear properties. Both strain gage rosettes and the modified extensometer were shown to be feasible shear strain transducers for the test method. The Iosipoescu shear test fixture was redesigned to incorporate several improvements. These improvements include accommodation of a 50 percent larger specimen for easier measurement of shear train, a clamping mechanism to relax strict tolerances on specimen width, and a self contained alignment tool for use during specimen installation. A set of in-plane and interlaminar shear properties were measured for three graphite fabric/epoxy composites of T300/934 composite material. The three weave patterns were Oxford, 5-harness satin, and 8-harness satin.

  17. Quasi-Static Viscoelastic Finite Element Model of an Aircraft Tire

    NASA Technical Reports Server (NTRS)

    Johnson, Arthur R.; Tanner, John A.; Mason, Angela J.

    1999-01-01

    An elastic large displacement thick-shell mixed finite element is modified to allow for the calculation of viscoelastic stresses. Internal strain variables are introduced at the element's stress nodes and are employed to construct a viscous material model. First order ordinary differential equations relate the internal strain variables to the corresponding elastic strains at the stress nodes. The viscous stresses are computed from the internal strain variables using viscous moduli which are a fraction of the elastic moduli. The energy dissipated by the action of the viscous stresses is included in the mixed variational functional. The nonlinear quasi-static viscous equilibrium equations are then obtained. Previously developed Taylor expansions of the nonlinear elastic equilibrium equations are modified to include the viscous terms. A predictor-corrector time marching solution algorithm is employed to solve the algebraic-differential equations. The viscous shell element is employed to computationally simulate a stair-step loading and unloading of an aircraft tire in contact with a frictionless surface.

  18. Molecular characterization of an unauthorized genetically modified Bacillus subtilis production strain identified in a vitamin B2 feed additive.

    PubMed

    Paracchini, Valentina; Petrillo, Mauro; Reiting, Ralf; Angers-Loustau, Alexandre; Wahler, Daniela; Stolz, Andrea; Schönig, Birgit; Matthies, Anastasia; Bendiek, Joachim; Meinel, Dominik M; Pecoraro, Sven; Busch, Ulrich; Patak, Alex; Kreysa, Joachim; Grohmann, Lutz

    2017-09-01

    Many food and feed additives result from fermentation of genetically modified (GM) microorganisms. For vitamin B2 (riboflavin), GM Bacillus subtilis production strains have been developed and are often used. The presence of neither the GM strain nor its recombinant DNA is allowed for fermentation products placed on the EU market as food or feed additive. A vitamin B 2 product (80% feed grade) imported from China was analysed. Viable B. subtilis cells were identified and DNAs of two bacterial isolates (LHL and LGL) were subjected to three whole genome sequencing (WGS) runs with different devices (MiSeq, 454 or HiSeq system). WGS data revealed the integration of a chloramphenicol resistance gene, the deletion of the endogenous riboflavin (rib) operon and presence of four putative plasmids harbouring rib operons. Event- and construct-specific real-time PCR methods for detection of the GM strain and its putative plasmids in food and feed products have been developed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Magnetic attachment for implant overdentures: influence of contact relationship with the denture base on stability and bending strain.

    PubMed

    Yang, Tsung-Chieh; Maeda, Yoshinobu; Gonda, Tomoya; Wada, Masahiro

    2013-01-01

    This study evaluated how the contact height between the magnetic attachment and denture base influences stability and bending strain. An implant modified with strain gauges and a magnetic attachment mounted in an acrylic resin block were used to characterize systems with varying degrees or heights of contact with the abutment. Bending strain under lateral loading increased significantly as the contact height decreased. In the no contact and resilient contact groups, magnetic assemblies separated at reduced bending strain in all loading conditions. The contact height of the magnetic attachment influenced the stability and the amount of bending strain on the implant.

  20. Strain Modulation of Electronic and Heat Transport Properties of Bilayer Boronitrene

    NASA Astrophysics Data System (ADS)

    Yang, Ming; Sun, Fang-Yuan; Wang, Rui-Ning; Zhang, Hang; Tang, Da-Wei

    2017-10-01

    Strain engineering has been proven as an effective approach to modify electronic and thermal properties of materials. Recently, strain effects on two-dimensional materials have become important relevant topics in this field. We performed density functional theory studies on the electronic and heat transport properties of bilayer boronitrene samples under an isotropic strain. We demonstrate that the strain will reduce the band gap width but keep the band gap type robust and direct. The strain will enhance the thermal conductivity of the system because of the increase in specific heat. The thermal conductivity was studied as a function of the phonon mean-free path.

  1. Investigation of carbon storage regulation network (csr genes) and phenotypic differences between acid sensitive and resistant Escherichia coli O157:H7 strains

    USDA-ARS?s Scientific Manuscript database

    Background: Escherichia coli O157:H7 and related serotype strains have previously been shown to vary in acid resistance, however, little is known about strain specific mechanisms of acid resistance. We examined sensitive and resistant E. coli strains to determine the effects of growth in minimal and...

  2. Synthesis and biological evaluation of asymmetric gramicidin S analogues containing modified D-phenylalanine residues.

    PubMed

    van der Knaap, Matthijs; Engels, Eefje; Busscher, Henk J; Otero, José M; Llamas-Saiz, Antonio L; van Raaij, Mark J; Mars-Groenendijk, Roos H; Noort, Daan; van der Marel, Gijsbert A; Overkleeft, Herman S; Overhand, Mark

    2009-09-01

    The synthesis of new analogues of the cationic antimicrobial peptide gramicidin S, having a modified D-phenylalanine residue, their antibacterial properties against several gram positive and negative strains, as well as their hemolytic activity is reported.

  3. Interaction of monopoles, dipoles, and turbulence with a shear flow

    NASA Astrophysics Data System (ADS)

    Marques Rosas Fernandes, V. H.; Kamp, L. P. J.; van Heijst, G. J. F.; Clercx, H. J. H.

    2016-09-01

    Direct numerical simulations have been conducted to examine the evolution of eddies in the presence of large-scale shear flows. The numerical experiments consist of initial-value-problems in which monopolar and dipolar vortices as well as driven turbulence are superposed on a plane Couette or Poiseuille flow in a periodic two-dimensional channel. The evolution of the flow has been examined for different shear rates of the background flow and different widths of the channel. Results found for retro-grade and pro-grade monopolar vortices are consistent with those found in the literature. Boundary layer vorticity, however, can significantly modify the straining and erosion of monopolar vortices normally seen for unbounded domains. Dipolar vortices are shown to be much more robust coherent structures in a large-scale shear flow than monopolar eddies. An analytical model for their trajectories, which are determined by self-advection and advection and rotation by the shear flow, is presented. Turbulent kinetic energy is effectively suppressed by the shearing action of the background flow provided that the shear is linear (Couette flow) and of sufficient strength. Nonlinear shear as present in the Poiseuille flow seems to even increase the turbulence strength especially for high shear rates.

  4. Strain accumulation in bituminous binders under repeated creep-recovery loading predicted from small-amplitude oscillatory shear (SAOS) experiments

    NASA Astrophysics Data System (ADS)

    Laukkanen, Olli-Ville; Winter, H. Henning

    2017-11-01

    The creep-recovery (CR) test starts out with a period of shearing at constant stress (creep) and is followed by a period of zero-shear stress where some of the accumulated shear strain gets reversed. Linear viscoelasticity (LVE) allows one to predict the strain response to repeated creep-recovery (RCR) loading from measured small-amplitude oscillatory shear (SAOS) data. Only the relaxation and retardation time spectra of a material need to be known and these can be determined from SAOS data. In an application of the Boltzmann superposition principle (BSP), the strain response to RCR loading can be obtained as a linear superposition of the strain response to many single creep-recovery tests. SAOS and RCR data were collected for several unmodified and modified bituminous binders, and the measured and predicted RCR responses were compared. Generally good agreement was found between the measured and predicted strain accumulation under RCR loading. However, in the case of modified binders, the strain accumulation was slightly overestimated (≤20% relative error) due to the insufficient SAOS information at long relaxation times. Our analysis also demonstrates that the evolution in the strain response under RCR loading, caused by incomplete recovery, can be reasonably well predicted by the presented methodology. It was also shown that the outlined modeling framework can be used, as a first approximation, to estimate the rutting resistance of bituminous binders by predicting the values of the Multiple Stress Creep Recovery (MSCR) test parameters.

  5. Mechanical properties of elytra from Tribolium castaneum wild-type and body color mutant strains

    USDA-ARS?s Scientific Manuscript database

    Cuticle tanning in insects involves simultaneous cuticular hardening and pigmentation. The dynamic mechanical properties of the highly modified and cuticle-rich forewings (elytra) from Tribolium castaneum (red flour beetle) body color mutant strains were investigated to determine the relationship b...

  6. Display of fungal hydrophobin on the Pichia pastoris cell surface and its influence on Candida antarctica lipase B

    PubMed Central

    Wang, Pan; He, Jie; Sun, Yufei; Reynolds, Matthew; Zhang, Li; Han, Shuangyan; Liang, Shuli; Sui, Haixin; Lin, Ying

    2016-01-01

    To modify the Pichia pastoris cell surface, two classes of hydrophobins, SC3 from Schizophyllum commune and HFBI from Trichoderma reesei, were separately displayed on the cell wall. There was an observable increase in the hydrophobicity of recombinant strains. Candida antarctica lipase B (CALB) was then co-displayed on the modified cells, generating strains GS115/SC3-61/CALB-51 and GS115/HFBI-61/CALB-51. Interestingly, the hydrolytic and synthetic activities of strain GS115/HFBI-61/CALB-51 increased by 37% and 109%, respectively, but decreased by 26% and 43%, respectively, in strain GS115/SC3-61/CALB-51 compared with the hydrophobin-minus recombinant strain GS115/CALB-GCW51. The amount of glycerol by-product from the transesterification reaction adsorbed on the cell surface was significantly decreased following hydrophobin modification, removing the glycerol barrier and allowing substrates to access the active sites of lipases. Electron micrographs indicated that the cell wall structures of both recombinant strains appeared altered, including changes to the inner glucan layer and outer mannan layer. These results suggest that the display of hydrophobins can change the surface structure and hydrophobic properties of P. pastoris, and affect the catalytic activities of CALB displayed on the surface of P. pastoris cells. PMID:26969039

  7. The Suppressor of AAC2 Lethality SAL1 Modulates Sensitivity of Heterologously Expressed Artemia ADP/ATP Carrier to Bongkrekate in Yeast

    PubMed Central

    Wysocka-Kapcinska, Monika; Torocsik, Beata; Turiak, Lilla; Tsaprailis, George; David, Cynthia L.; Hunt, Andrea M.; Vekey, Karoly; Adam-Vizi, Vera; Kucharczyk, Roza; Chinopoulos, Christos

    2013-01-01

    The ADP/ATP carrier protein (AAC) expressed in Artemia franciscana is refractory to bongkrekate. We generated two strains of Saccharomyces cerevisiae where AAC1 and AAC3 were inactivated and the AAC2 isoform was replaced with Artemia AAC containing a hemagglutinin tag (ArAAC-HA). In one of the strains the suppressor of ΔAAC2 lethality, SAL1, was also inactivated but a plasmid coding for yeast AAC2 was included, because the ArAACΔsal1Δ strain was lethal. In both strains ArAAC-HA was expressed and correctly localized to the mitochondria. Peptide sequencing of ArAAC expressed in Artemia and that expressed in the modified yeasts revealed identical amino acid sequences. The isolated mitochondria from both modified strains developed 85% of the membrane potential attained by mitochondria of control strains, and addition of ADP yielded bongkrekate-sensitive depolarizations implying acquired sensitivity of ArAAC-mediated adenine nucleotide exchange to this poison, independent from SAL1. However, growth of ArAAC-expressing yeasts in glycerol-containing media was arrested by bongkrekate only in the presence of SAL1. We conclude that the mitochondrial environment of yeasts relying on respiratory growth conferred sensitivity of ArAAC to bongkrekate in a SAL1-dependent manner. PMID:24073201

  8. Triacetic acid lactone production in industrial Saccharomyces yeast strains

    USDA-ARS?s Scientific Manuscript database

    Triacetic acid lactone (TAL) is a potential platform chemical that can be produced in yeast. To evaluate the potential for industrial yeast strains to produce TAL, the g2ps1 gene encoding 2-pyrone synthase was transformed into thirteen industrial yeast strains of varied genetic background. TAL produ...

  9. Anaplasma marginale superinfection attributable to pathogen strains with distinct genomic backgrounds.

    USDA-ARS?s Scientific Manuscript database

    Microbial strain structure is dynamic over space and time; shifts in pathogen strain structure result in changing patterns of disease. The scale of change in space and time differs markedly among pathogens depending on multiple factors including pathogen-specific mechanisms of genetic change and the...

  10. Engineering industrial Saccharomyces cerevisiae strains for xylose fermentation and comparison for switchgrass conversion

    USDA-ARS?s Scientific Manuscript database

    Saccharomyces physiology and fermentation related properties vary broadly among industrial strains. In this study, six industrial strains of varied genetic background were engineered to ferment xylose. Aerobic growth rates on xylose were 0.040 h**-1 to 0.167 h**-1. Fermentation of xylose, glucose/xy...

  11. A novel multigene expression construct for modification of glycerol metabolism in Yarrowia lipolytica

    PubMed Central

    2013-01-01

    Background High supply of raw, residual glycerol from biodiesel production plants promote the search for novel biotechnological methods of its utilization. In this study we attempted modification of glycerol catabolism in a nonconventional yeast species Yarrowia lipolytica through genetic engineering approach. Results To address this, we developed a novel genetic construct which allows transferring three heterologous genes, encoding glycerol dehydratase, its reactivator and a wide-spectrum alcohol oxidoreductase under the control of glycerol-induced promoter. The three genes, tandemly arrayed in an expression cassette with a marker gene ura3, regulatory and targeting sequences (G3P dh promoter and XPR-like terminator, 28S rDNA as a target locus), were transferred into Yarrowia lipolytica cells. The obtained recombinant strain NCYC3825 was characterized at the molecular level and with respect to its biotechnological potential. Our experiments indicated that the novel recombinant strain stably borne one copy of the expression cassette and efficiently expressed heterologous alcohol oxidoreductase, while glycerol dehydratase and its reactivator were expressed at lower level. Comparative shake flask cultivations in glucose- and glycerol-based media demonstrated higher biomass production by the recombinant strain when glycerol was the main carbon source. During bioreactor (5 L) fed-batch cultivation in glycerol-based medium, the recombinant strain was characterized by relatively high biomass and lipids accumulation (up to 42 gDCW L-1, and a peak value of 38%LIPIDS of DCW, respectively), and production of high titers of citric acid (59 g L-1) and 2-phenylethanol (up to 1 g L-1 in shake flask cultivation), which are industrially attractive bioproducts. Conclusions Due to heterogeneous nature of the observed alterations, we postulate that the main driving force of the modified phenotype was faster growth in glycerol-based media, triggered by modifications in the red-ox balance brought by the wide spectrum oxidoreductase. Our results demonstrate the potential multidirectional use of a novel Yarrowia lipolytica strain as a microbial cell factory. PMID:24188724

  12. UDP-4-Keto-6-Deoxyglucose, a Transient Antifungal Metabolite, Weakens the Fungal Cell Wall Partly by Inhibition of UDP-Galactopyranose Mutase

    PubMed Central

    Ma, Liang; Salas, Omar; Bowler, Kyle

    2017-01-01

    ABSTRACT Can accumulation of a normally transient metabolite affect fungal biology? UDP-4-keto-6-deoxyglucose (UDP-KDG) represents an intermediate stage in conversion of UDP-glucose to UDP-rhamnose. Normally, UDP-KDG is not detected in living cells, because it is quickly converted to UDP-rhamnose by the enzyme UDP-4-keto-6-deoxyglucose-3,5-epimerase/-4-reductase (ER). We previously found that deletion of the er gene in Botrytis cinerea resulted in accumulation of UDP-KDG to levels that were toxic to the fungus due to destabilization of the cell wall. Here we show that these negative effects are at least partly due to inhibition by UDP-KDG of the enzyme UDP-galactopyranose mutase (UGM), which reversibly converts UDP-galactopyranose (UDP-Galp) to UDP-galactofuranose (UDP-Galf). An enzymatic activity assay showed that UDP-KDG inhibits the B. cinerea UGM enzyme with a Ki of 221.9 µM. Deletion of the ugm gene resulted in strains with weakened cell walls and phenotypes that were similar to those of the er deletion strain, which accumulates UDP-KDG. Galf residue levels were completely abolished in the Δugm strain and reduced in the Δer strain, while overexpression of the ugm gene in the background of a Δer strain restored Galf levels and alleviated the phenotypes. Collectively, our results show that the antifungal activity of UDP-KDG is due to inhibition of UGM and possibly other nucleotide sugar-modifying enzymes and that the rhamnose metabolic pathway serves as a shunt that prevents accumulation of UDP-KDG to toxic levels. These findings, together with the fact that there is no Galf in mammals, support the possibility of developing UDP-KDG or its derivatives as antifungal drugs. PMID:29162710

  13. Differences in extinction of conditioned fear in C57BL/6 substrains are unrelated to expression of alpha-synuclein.

    PubMed

    Siegmund, Anja; Langnaese, Kristina; Wotjak, Carsten T

    2005-02-28

    C57BL/6 mice are commonly used as background strains for genetically modified mice, and little attention is usually paid to the notification of the specific substrain. However, it is known that C57BL/6NCrl (B6N) and C57BL/6JOlaHsd (B6JOla) mice differ in the course of extinction of conditioned fear (Stiedl O, Radulovic J, Lohmann R, Birkenfeld K, Palve M, Kammermeier J, et al. Strain and substrain differences in context- and tone-dependent fear conditioning of inbred mice. Behav Brain Res 1999;104:1-12), as well as in the expression of alpha-synuclein (Specht CG, Schoepfer R. Deletion of the alpha-synuclein locus in a subpopulation of C57BL/6J inbred mice. BMC Neurosci 2001;2:11). We tested for a causal relationship between the two findings by employing B6N (expressing alpha-synuclein), B6JOla (not expressing alpha-syn) and the third strain C57BL/6JCrl (B6Jax, expressing alpha-syn). We show that alpha-syn does not account for differences in extinction in a fear conditioning task, as its expression did not covary with the decrease of freezing on repeated non-reinforced tone and context exposure in the three strains: B6Jax exhibited fastest extinction followed by B6JOla. In contrast, B6N showed persistent fear over the course of extinction training. The differences in extinction between B6JOla and B6N were unrelated to sensorimotor processing (pain threshold and basal tone reaction) and innate fear (light-dark test). However, B6Jax displayed less innate fear than B6JOla and B6N. Our results of marked differences in innate and conditioned fear in three B6 substrains illustrate the necessity of a strict adherence to an exact mouse strain nomenclature.

  14. Constraining Modified Theories of Gravity with Gravitational-Wave Stochastic Backgrounds

    NASA Astrophysics Data System (ADS)

    Maselli, Andrea; Marassi, Stefania; Ferrari, Valeria; Kokkotas, Kostas; Schneider, Raffaella

    2016-08-01

    The direct discovery of gravitational waves has finally opened a new observational window on our Universe, suggesting that the population of coalescing binary black holes is larger than previously expected. These sources produce an unresolved background of gravitational waves, potentially observable by ground-based interferometers. In this Letter we investigate how modified theories of gravity, modeled using the parametrized post-Einsteinian formalism, affect the expected signal, and analyze the detectability of the resulting stochastic background by current and future ground-based interferometers. We find the constraints that Advanced LIGO would be able to set on modified theories, showing that they may significantly improve the current bounds obtained from astrophysical observations of binary pulsars.

  15. [Comparison of the quick Gram stain method to the B&M modified and favor methods].

    PubMed

    Osawa, Kayo; Kataoka, Nobumasa; Maruo, Toshio

    2011-01-01

    The Gram stain is an established method for bacterial identification, but the time needed to carry out this stain is 2-3 min. We attempted to shorten this time and stained a total of 70 clinical specimens isolated from using the Bartholomew & Mittwer (B&M) modified or Favor methods with a 3 s duration for washing and staining steps. Results were plotted and analyzed using a Hue Saturation Intensity (HSI) model. The range based on a plot of the two methods with the HSI model was presented as a reference interval. Our results indicated that 100% (35/35) of strains were Gram positive and 97.1% (34/35) were Gram negative for the quick B&M modified method. In the quick Favor method, 80.0% (28/35) were Gram positive and 68.6% (24/35) of strains were Gram negative. We propose that the quick B&M modified method is equivalent to the standard Gram staining method and is superior to the quick Favor method.

  16. Completion of the swine genome will simplify the production of swine as a large animal biomedical model

    PubMed Central

    2012-01-01

    Background Anatomic and physiological similarities to the human make swine an excellent large animal model for human health and disease. Methods Cloning from a modified somatic cell, which can be determined in cells prior to making the animal, is the only method available for the production of targeted modifications in swine. Results Since some strains of swine are similar in size to humans, technologies that have been developed for swine can be readily adapted to humans and vice versa. Here the importance of swine as a biomedical model, current technologies to produce genetically enhanced swine, current biomedical models, and how the completion of the swine genome will promote swine as a biomedical model are discussed. Conclusions The completion of the swine genome will enhance the continued use and development of swine as models of human health, syndromes and conditions. PMID:23151353

  17. Sterilization of Bacillus pumilus spores using supercritical fluid carbon dioxide containing various modifier solutions.

    PubMed

    Shieh, Edison; Paszczynski, Andrzej; Wai, Chien M; Lang, Qingyong; Crawford, Ronald L

    2009-03-01

    Supercritical fluid carbon dioxide (SF-CO(2)) with small amounts of chemical modifier(s) provides a very effective sterilization technique that should be useful for destroying microorganism on heat-sensitive devices such as instruments flown on planetary-bound spacecraft. Under a moderate temperature (50 degrees C) and pressure (100 atm), spores of Bacillus pumilus strains ATCC 7061 and SAFR 032 can be effectively inactivated/eliminated from metal surfaces and small electronic devices in only 45 min using optimized modifier concentrations. Modifiers explored in this study included hydrogen peroxide (H(2)O(2)), tert-butyl hydroperoxide, formic acid, and Triton X-100. During sterilization procedure the modifiers were continuously added to SF-CO(2) in either methanol or water at controlled concentrations. The lowest effective concentrations were established for each modifier. Complete elimination of both types of B. pumilus endospores occurred with an optimal modifier addition of either or 10% methanol containing 12% H(2)O(2) or 12% tert-butyl hydroperoxide in SF-CO(2), or a mixture of 6% H(2)O(2) and 6% tert-butyl hydroperoxide. Using water as the carrier of SF-CO(2) modifier, the complete elimination of spores viability of both B. pumilus strains occurred with an addition of either 3.3% water containing 3% H(2)O(2), or 3.3% water containing 10% methanol and 0.5% formic acid, or 3.3% water containing 10% methanol, 1% formic acid and 2% H(2)O(2).

  18. Combining inhibitor tolerance and D-xylose fermentation in industrial Saccharomyces cerevisiae for efficient lignocellulose-based bioethanol production

    PubMed Central

    2013-01-01

    Background In addition to efficient pentose utilization, high inhibitor tolerance is a key trait required in any organism used for economically viable industrial bioethanol production with lignocellulose biomass. Although recent work has succeeded in establishing efficient xylose fermentation in robust industrial Saccharomyces cerevisiae strains, the resulting strains still lacked sufficient inhibitor tolerance for efficient sugar fermentation in lignocellulose hydrolysates. The aim of the present work was to combine high xylose fermentation activity and high inhibitor tolerance in a single industrial yeast strain. Results We have screened 580 yeast strains for high inhibitor tolerance using undetoxified acid-pretreated spruce hydrolysate and identified a triploid industrial baker’s yeast strain as having the highest inhibitor tolerance. From this strain, a mating competent diploid segregant with even higher inhibitor tolerance was obtained. It was crossed with the recently developed D-xylose fermenting diploid industrial strain GS1.11-26, with the Ethanol Red genetic background. Screening of 819 diploid segregants from the tetraploid hybrid resulted in two strains, GSF335 and GSF767, combining high inhibitor tolerance and efficient xylose fermentation. In a parallel approach, meiotic recombination of GS1.11-26 with a haploid segregant of Ethanol Red and screening of 104 segregants resulted in a similar inhibitor tolerant diploid strain, GSE16. The three superior strains exhibited significantly improved tolerance to inhibitors in spruce hydrolysate, higher glucose consumption rates, higher aerobic growth rates and higher maximal ethanol accumulation capacity in very-high gravity fermentation, compared to GS1.11-26. In complex medium, the D-xylose utilization rate by the three superior strains ranged from 0.36 to 0.67 g/g DW/h, which was lower than that of GS1.11-26 (1.10 g/g DW/h). On the other hand, in batch fermentation of undetoxified acid-pretreated spruce hydrolysate, the three superior strains showed comparable D-xylose utilization rates as GS1.11-26, probably because of their higher inhibitor tolerance. They produced up to 23% more ethanol compared to Ethanol Red. Conclusions We have successfully constructed three superior industrial S. cerevisiae strains that combine efficient D-xylose utilization with high inhibitor tolerance. Since the background strain Ethanol Red has a proven record of successful industrial application, the three new superior strains have strong potential for direct application in industrial bioethanol production. PMID:23971950

  19. Clostridium botulinum strains producing BoNT/F4 or BoNT/F5.

    PubMed

    Raphael, Brian H; Bradshaw, Marite; Kalb, Suzanne R; Joseph, Lavin A; Lúquez, Carolina; Barr, John R; Johnson, Eric A; Maslanka, Susan E

    2014-05-01

    Botulinum neurotoxin type F (BoNT/F) may be produced by Clostridium botulinum alone or in combination with another toxin type such as BoNT/A or BoNT/B. Type F neurotoxin gene sequences have been further classified into seven toxin subtypes. Recently, the genome sequence of one strain of C. botulinum (Af84) was shown to contain three neurotoxin genes (bont/F4, bont/F5, and bont/A2). In this study, eight strains containing bont/F4 and seven strains containing bont/F5 were examined. Culture supernatants produced by these strains were incubated with BoNT/F-specific peptide substrates. Cleavage products of these peptides were subjected to mass spectral analysis, allowing detection of the BoNT/F subtypes present in the culture supernatants. PCR analysis demonstrated that a plasmid-specific marker (PL-6) was observed only among strains containing bont/F5. Among these strains, Southern hybridization revealed the presence of an approximately 242-kb plasmid harboring bont/F5. Genome sequencing of four of these strains revealed that the genomic backgrounds of strains harboring either bont/F4 or bont/F5 are diverse. None of the strains analyzed in this study were shown to produce BoNT/F4 and BoNT/F5 simultaneously, suggesting that strain Af84 is unusual. Finally, these data support a role for the mobility of a bont/F5-carrying plasmid among strains of diverse genomic backgrounds.

  20. Complete Sequence of the Intronless Mitochondrial Genome of the Saccharomyces cerevisiae Strain CW252

    PubMed Central

    2018-01-01

    ABSTRACT The mitochondrial genomes of Saccharomyces cerevisiae strains contain up to 13 introns. An intronless recombinant genome introduced into the nuclear background of S. cerevisiae strain W303 gave the S. cerevisiae CW252 strain, which is used to model mitochondrial respiratory pathologies. The complete sequence of this mitochondrial genome was obtained using a hybrid assembling methodology. PMID:29700138

  1. Corticosterone facilitates extinction of fear memory in BALB/c mice but strengthens cue related fear in C57BL/6 mice.

    PubMed

    Brinks, V; de Kloet, E R; Oitzl, M S

    2009-04-01

    Corticosterone, the naturally occurring glucocorticoid of rodents is secreted in response to stressors and is known for its facilitating and detrimental effects on emotional learning and memory. The large variability in the action of corticosterone on processing of emotional memories is postulated to depend on genetic background and the spatio-temporal domain in which the hormone operates. To address this hypothesis, mice of two strains with distinct corticosterone secretory patterns and behavioural phenotype (BALB/c and C57BL/6J) were treated with corticosterone (250 microg/kg, i.p.), either 5 min before or directly after acquisition in a fear conditioning task. As the paradigm allowed assessing in one experimental procedure both context- and cue-related fear behaviour, we were able to detect generalization and specificity of fear. BALB/c showed generalized strong fear memory, while C57BL/6J mice discriminated between freezing during context- and cue episodes. Corticosterone had opposite effects on fear memory depending on the strain and time of injection. Corticosterone after acquisition did not affect C57BL/6J mice, but destabilized consolidation and facilitated extinction in BALB/c. Corticosterone 5 min before acquisition strengthened stress-associated signals: BALB/c no longer showed lower fear memory, while C57BL/6J mice displayed increased fear memory and impaired extinction in cue episodes. We propose that corticosterone-induced facilitation of fear memory in C57BL/6J mice can be used to study the development of fear memories, corticosterone administration in BALB/c mice presents a model to examine treatment. We conclude that genetic background and time of corticosterone action are modifiers of fear memory with interesting translational implications for anxiety-related diseases.

  2. Inhibition of Actinobacillus actinomycetemcomitans leukotoxicity by bacteria from the subgingival flora.

    PubMed

    Johansson, A; Hänström, L; Kalfas, S

    2000-08-01

    Actinobacillus actinomycetemcomitans produces a pore-forming leukotoxin that lyses human polymorphonuclear leukocytes and monocytes. Certain proteolytic bacteria may coexist with A. actinomycetemcomitans in periodontal pockets. We aimed therefore to examine whether oral bacteria can modify the leukotoxicity of A. actinomycetemcomitans. A total of 55 strains representing 45 bacterial species of the subgingival flora were tested. Each strain was incubated with the highly toxic strain of A. actinomycetemcomitans HK 1519 and the leukotoxic activity of the suspension against human polymorphonuclear leukocytes was determined from the activity of the lactate dehydrogenase released upon lysis of the leukocytes. Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica and Prevotella loeschii inhibited the leukotoxicity of A. actinomycetemcomitans cells as well as the activity of leukotoxin purified from the same strain. The bacterial strains without the ability to block leukotoxic activity also failed to destroy pure leukotoxin even after 5 h of incubation. The proteolytic degradation of leukotoxin by P. gingivalis was mainly dependent on the activity of the enzymes R- and K-gingipains. P. intermedia and P. nigrescens also degraded the leukotoxin by enzymes. The results imply a role of the periodontal microflora in modifying the virulence of A. actinomycetemcomitans by destroying its leukotoxin.

  3. Acrolein inhalation causes myocardial strain delay and decreased cardiac performance as detected by high-frequency echocardiography in mice

    EPA Science Inventory

    Acrolein, an unsaturated aldehyde found in air pollution, impairs Ca2+ flux and contraction in cardiomyocytes in vitro. To better define direct and delayed functional cardiac effects, we hypothesized that a single exposure to acrolein would modify myocardial strain and performanc...

  4. Sensitivity of infectious bovine rhinotracheitis virus to ether.

    PubMed

    Crandell, R A; Melloh, A J; Sorlie, P

    1975-12-01

    The sensitivity of 12 field isolates of infectious boviine rhinotracheitis (IBR) virus and four commercial modified-live infectious bovine rhinotracheitis virus vaccine strains was determined after exposure to 20% ethyl ether (anesthetic) for 16 h at 4 c. The infectivity of five field strains was reduced by varying degrees, whereas seven were found to be resistant. Three vaccine strains were moderately sensitive, and one strain was resistant. Four of the sensitive field strains were isolated from the conjunctiva and the other was isolated from the surface of the epiglottis of natural infective cattle. Strains of virus isolated from fetal tissue and nasal cavity were resistant. All viruses were readily inactivated by chloroform treatment.

  5. Differences in genetic background influence the induction of innate and acquired immune responses in chickens depending on the virulence of the infecting infectious bursal disease virus (IBDV) strain.

    PubMed

    Aricibasi, Merve; Jung, Arne; Heller, E Dan; Rautenschlein, Silke

    2010-05-15

    Previous studies and field observations have suggested that genetic background influences infectious bursal disease virus (IBDV) pathogenesis. However, the influence of the virulence of the infecting IBDV strain and the mechanisms underlying the differences in susceptibility are not known. In the present study IBDV pathogenesis was compared between specific-pathogen-free layer-type (LT) chickens, which are the most susceptible chicken for IBDV and have been used as the model for pathogenesis studies, and broiler-type (BT) chickens, which are known to be less susceptible to clinical infectious bursal disease (IBD). The innate and acquired immune responses were investigated after inoculation of an intermediate (i), virulent (v) or very virulent (vv) strain of IBDV. IBDV pathogenesis was comparable among genetic backgrounds after infection with iIBDV. After infection with vIBDV and vvIBDV, LT birds showed severe clinical disease and mortality, higher bursal lesion scores and IBDV-antigen load relative to BT birds. Circulating cytokine induction varied significantly in both timing and quantity between LT and BT birds and among virus strains (P<0.05). Evaluation of different immune cell populations by flow-cytometric analysis in the bursa of Fabricius provided circumstantial evidence of a stronger local T cell response in BT birds vs. LT birds after infection with the virulent strain. On the other hand, LT birds showed a more significant increase in circulating macrophage-derived immune mediators such as total interferon (IFN) and serum nitrite than BT birds on days 2 and 3 post-vIBDV infection (P<0.05). Stronger stimulation of innate immune reactions especially after vIBDV infection in the early phase may lead to faster and more severe lesion development accompanied by clinical disease and death in LT chickens relative to BT chickens. Interestingly, no significant differences were seen between genetic backgrounds in induction of the IBDV-specific humoral response: timing of IBDV-antibody induction and antibody levels were comparable between BT and LT birds. This study clearly demonstrates a significant influence of chickens' genetic background on disease outcome. The difference between backgrounds in IBDV susceptibility is further influenced by the virulence of the infecting virus strain. Copyright 2009 Elsevier B.V. All rights reserved.

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

    Dai, Ziyu; Aryal, Uma K.; Shukla, Anil

    ALG3 is a Family 58 glycosyltransferase enzyme involved in early N-linked glycan synthesis. Here, we investigated the effect of the alg3 gene disruption on growth, development, metabolism, and protein secretion in Aspergillus niger. The alg3 gene deletion resulted in a significant reduction of growth on complete (CM) and potato dextrose agar (PDA) media and a substantial reduction of spore production on CM. It also delayed spore germination in the liquid cultures of both CM and PDA media, but led to a significant accumulation of red pigment on both CM and liquid modified minimal medium (MM) supplemented with yeast extract. Themore » relative abundance of 55 proteins of the total 190 proteins identified in the secretome was significantly different as a result of alg3 gene deletion. Comparison of a Trichoderma reesei cellobiohydrolase (Cel7A) heterologously expressed in A. niger parental and Δalg3 strains showed that the recombinant Cel7A expressed in the mutant background was smaller in size than that from the parental strains. This study suggests that ALG3 is critical for growth and development, pigment production, and protein secretion in A. niger. Functional analysis of recombinant Cel7A with aberrant glycosylation demonstrates the feasibility of this alternative approach to evaluate the role of N-linked glycosylation in glycoprotein secretion and function.« less

  7. Fluorogenic Strain-Promoted Alkyne-Diazo Cycloadditions.

    PubMed

    Friscourt, Frédéric; Fahrni, Christoph J; Boons, Geert-Jan

    2015-09-28

    Fluorogenic reactions, in which non- or weakly fluorescent reagents produce highly fluorescent products, are attractive for detecting a broad range of compounds in the fields of bioconjugation and material sciences. Herein, we report that a dibenzocyclooctyne derivative modified with a cyclopropenone moiety (Fl-DIBO) can undergo fast strain-promoted cycloaddition reactions under catalyst-free conditions with azides, nitrones, nitrile oxides, as well as mono- and disubstituted diazo-derivatives. Although the reaction with nitrile oxides, nitrones, and disubstituted diazo compounds gave cycloadducts with low quantum yield, monosubstituted diazo reagents produced 1H-pyrazole derivatives that exhibited an approximately 160-fold fluorescence enhancement over Fl-DIBO combined with a greater than 10,000-fold increase in brightness. Concluding from quantum chemical calculations, fluorescence quenching of 3H-pyrazoles, which are formed by reaction with disubstituted diazo-derivatives, is likely due to the presence of energetically low-lying (n,π*) states. The fluorogenic probe Fl-DIBO was successfully employed for the labeling of diazo-tagged proteins without detectable background signal. Diazo-derivatives are emerging as attractive reporters for the labeling of biomolecules, and the studies presented herein demonstrate that Fl-DIBO can be employed for visualizing such biomolecules without the need for probe washout. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Full-Genome Sequencing Identifies in the Genetic Background Several Determinants That Modulate the Resistance Phenotype in Methicillin-Resistant Staphylococcus aureus Strains Carrying the Novel mecC Gene

    PubMed Central

    de Lencastre, Hermínia; Tomasz, Alexander

    2017-01-01

    ABSTRACT Most methicillin-resistant Staphylococcus aureus (MRSA) strains are resistant to beta-lactam antibiotics due to the presence of the mecA gene, encoding an extra penicillin-binding protein (PBP2A) that has low affinity for virtually all beta-lactam antibiotics. Recently, a new resistance determinant—the mecC gene—was identified in S. aureus isolates recovered from humans and dairy cattle. Although having typically low MICs to beta-lactam antibiotics, MRSA strains with the mecC determinant are also capable of expressing high levels of oxacillin resistance when in an optimal genetic background. In order to test the impact of extensive beta-lactam selection on the emergence of mecC-carrying strains with high levels of antibiotic resistance, we exposed the prototype mecC-carrying MRSA strain, LGA251, to increasing concentrations of oxacillin. LGA251 was able to rapidly adapt to high concentrations of oxacillin in growth medium. In such laboratory mutants with increased levels of oxacillin resistance, we identified mutations in genes with no relationship to the mecC regulatory system, indicating that the genetic background plays an important role in the establishment of the levels of oxacillin resistance. Our data also indicate that the stringent stress response plays a critical role in the beta-lactam antibiotic resistance phenotype of MRSA strains carrying the mecC determinant. PMID:28069659

  9. Genome-Wide Gene Expression Analysis of Bordetella pertussis Isolates Associated with a Resurgence in Pertussis: Elucidation of Factors Involved in the Increased Fitness of Epidemic Strains

    PubMed Central

    King, Audrey J.; van der Lee, Saskia; Mohangoo, Archena; van Gent, Marjolein; van der Ark, Arno; van de Waterbeemd, Bas

    2013-01-01

    Bordetella pertussis (B. pertussis) is the causative agent of whooping cough, which is a highly contagious disease in the human respiratory tract. Despite vaccination since the 1950s, pertussis remains the most prevalent vaccine-preventable disease in developed countries. A recent resurgence pertussis is associated with the expansion of B. pertussis strains with a novel allele for the pertussis toxin (ptx) promoter ptxP3 in place of resident ptxP1 strains. The recent expansion of ptxP3 strains suggests that these strains carry mutations that have increased their fitness. Compared to the ptxP1 strains, ptxP3 strains produce more Ptx, which results in increased virulence and immune suppression. In this study, we investigated the contribution of gene expression changes of various genes on the increased fitness of the ptxP3 strains. Using genome-wide gene expression profiling, we show that several virulence genes had higher expression levels in the ptxP3 strains compared to the ptxP1 strains. We provide the first evidence that wildtype ptxP3 strains are better colonizers in an intranasal mouse infection model. This study shows that the ptxP3 mutation and the genetic background of ptxP3 strains affect fitness by contributing to the ability to colonize in a mouse infection model. These results show that the genetic background of ptxP3 strains with a higher expression of virulence genes contribute to increased fitness. PMID:23776625

  10. A quantitative survey of gravity receptor function in mutant mouse strains.

    PubMed

    Jones, Sherri M; Johnson, Kenneth R; Yu, Heping; Erway, Lawrence C; Alagramam, Kumar N; Pollak, Natasha; Jones, Timothy A

    2005-12-01

    The purpose of this research was to identify vestibular deficits in mice using linear vestibular evoked potentials (VsEPs). VsEP thresholds, peak latencies, and peak amplitudes from 24 strains with known genetic mutations and 6 inbred background strains were analyzed and descriptive statistics generated for each strain. Response parameters from mutant homozygotes were compared with heterozygote and/or background controls and all strain averages were contrasted to normative ranges. Homozygotes of the following recessive mutations had absent VsEPs at the ages tested: Espn(je), Atp2b2dfw-2J, Spnb4qv-lnd2J, Spnb4qv-3J, Myo7ash1, Tmie(sr), Myo6sv, jc, Pcdh15av-J, Pcdh15av-2J, Pcdh15av-3J, Cdh23v-2J, Sans(js), hr, Kcne1pkr and Pou3f4del. These results suggest profound gravity receptor deficits for these homozygotes, which is consistent with the structural deficits that have been documented for many of these strains. Homozygotes of Catna2cdf, Grid2ho4J, Wnt1sw, qk, and Mbpshi strains and heterozygotes of Grid2lc had measurable VsEPs but one or more response parameters differed from the respective control group (heterozygote or background strain) or were outside normal ranges. For example, qk and Mbpshi homozygotes showed significantly prolonged latencies consistent with the abnormal myelin that has been described for these strains. Prolonged latencies may suggest deficits in neural conduction; elevated thresholds suggest reduced sensitivity, and reduced amplitudes may be suggestive for reduced neural synchrony. One mutation, Otx1jv, had all VsEP response parameters within normal limits--an expected finding because the abnormality in Otxljv is presumably restricted to the lateral semicircular canal. Interestingly, some heterozygote groups also showed abnormalities in one or more VsEP response parameters, suggesting that vestibular dysfunction, although less severe, may be present in some heterozygous animals.

  11. Nonthmicin, a Polyether Polyketide Bearing a Halogen-Modified Tetronate with Neuroprotective and Antiinvasive Activity from Actinomadura sp.

    PubMed

    Igarashi, Yasuhiro; Matsuoka, Noriaki; In, Yasuko; Kataura, Tetsushi; Tashiro, Etsu; Saiki, Ikuo; Sudoh, Yuri; Duangmal, Kannika; Thamchaipenet, Arinthip

    2017-03-17

    Nonthmicin (1), a new polyether polyketide bearing a chlorinated tetronic acid, was isolated from the culture extract of a soil-derived Actinomadura strain. The structure of 1 was elucidated by interpretation of NMR and MS spectroscopic data, and the absolute configuration of 1 was proposed on the basis of the crystal structure of its dechloro congener ecteinamycin (2) also isolated from the same strain. Tetronic acids modified by halogenation have never been reported from natural products. Compounds 1 and 2 were found to have neuroprotective activity and antimetastatic properties at submicromolar concentrations in addition to antibacterial activity.

  12. An azole-resistant isolate of Malassezia pachydermatis.

    PubMed

    Nijima, Misako; Kano, Rui; Nagata, Masahiko; Hasegawa, Atsuhiko; Kamata, Hiroshi

    2011-04-21

    Canine Malassezia dermatitis (MD) is frequently treated with systemic ketoconazole (KTZ) and itaconazole (ITZ). However, the antifungal susceptibility of clinical isolates of M. pachydermatis from dogs and cats to the azoles has not been well investigated. In the present study, the in vitro susceptibility of the standard strain (CBS1879: the neotype strain of M. pachydermatis) and 29 clinical isolates of M. pachydermatis to the azoles was measured by a modified CLSI M27-A2 test using modified Dixon medium as well as by the E-test. The minimum inhibitory concentrations (MICs) of the 30 isolates of M. pachydermatis (including the neotype strain) against KTZ and ITZ were <0.03 μg/ml by the two methods. The MICs of 1 clinical isolate (ASC-11) were 1 and 2 μg/ml against KTZ, and 2 and 8 μg/ml against ITZ, by the modified CLSI M27-A2 test and the E-test, respectively. Thus, isolate ASC-11 may be resistant to these azoles, making this the first report of a resistant isolate of M. pachydermatis to KTZ and ITZ. Copyright © 2010 Elsevier B.V. All rights reserved.

  13. Modeling the Nonlinear, Strain Rate Dependent Deformation of Woven Ceramic Matrix Composites With Hydrostatic Stress Effects Included

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.; Carney, Kelly S.

    2004-01-01

    An analysis method based on a deformation (as opposed to damage) approach has been developed to model the strain rate dependent, nonlinear deformation of woven ceramic matrix composites with a plain weave fiber architecture. In the developed model, the differences in the tension and compression response have also been considered. State variable based viscoplastic equations originally developed for metals have been modified to analyze the ceramic matrix composites. To account for the tension/compression asymmetry in the material, the effective stress and effective inelastic strain definitions have been modified. The equations have also been modified to account for the fact that in an orthotropic composite the in-plane shear stiffness is independent of the stiffness in the normal directions. The developed equations have been implemented into a commercially available transient dynamic finite element code, LS-DYNA, through the use of user defined subroutines (UMATs). The tensile, compressive, and shear deformation of a representative plain weave woven ceramic matrix composite are computed and compared to experimental results. The computed values correlate well to the experimental data, demonstrating the ability of the model to accurately compute the deformation response of woven ceramic matrix composites.

  14. Valyl-tRNA synthetase modification-dependent restriction of bacteriophage T4.

    PubMed Central

    Olson, N J; Marchin, G L

    1984-01-01

    A strain of Escherichia coli, CP 790302, severely restricts the growth of wild-type bacteriophage T4. In broth culture, most infections of single cells are abortive, although a few infected cells exhibit reduced burst sizes. In contrast, bacteriophage T4 mutants impaired in the ability to modify valyl-tRNA synthetase develop normally on this strain. Biochemical evidence indicates that the phage-modified valyl-tRNA synthetase in CP 790302 is different from that previously described. Although the enzyme is able to support normal protein synthesis, a disproportionate amount of phage structural protein (serum blocking power) fails to mature into particles of the appropriate density. The results with host strain CP 790302 are consistent with either a gratuitous inhibition of phage assembly by faulty modification or abrogation of an unknown role that valyl-tRNA synthetase might normally play in viral assembly. PMID:6374167

  15. An efficient genotyping method for genome-modified animals and human cells generated with CRISPR/Cas9 system.

    PubMed

    Zhu, Xiaoxiao; Xu, Yajie; Yu, Shanshan; Lu, Lu; Ding, Mingqin; Cheng, Jing; Song, Guoxu; Gao, Xing; Yao, Liangming; Fan, Dongdong; Meng, Shu; Zhang, Xuewen; Hu, Shengdi; Tian, Yong

    2014-09-19

    The rapid generation of various species and strains of laboratory animals using CRISPR/Cas9 technology has dramatically accelerated the interrogation of gene function in vivo. So far, the dominant approach for genotyping of genome-modified animals has been the T7E1 endonuclease cleavage assay. Here, we present a polyacrylamide gel electrophoresis-based (PAGE) method to genotype mice harboring different types of indel mutations. We developed 6 strains of genome-modified mice using CRISPR/Cas9 system, and utilized this approach to genotype mice from F0 to F2 generation, which included single and multiplexed genome-modified mice. We also determined the maximal detection sensitivity for detecting mosaic DNA using PAGE-based assay as 0.5%. We further applied PAGE-based genotyping approach to detect CRISPR/Cas9-mediated on- and off-target effect in human 293T and induced pluripotent stem cells (iPSCs). Thus, PAGE-based genotyping approach meets the rapidly increasing demand for genotyping of the fast-growing number of genome-modified animals and human cell lines created using CRISPR/Cas9 system or other nuclease systems such as TALEN or ZFN.

  16. An Efficient Genotyping Method for Genome-modified Animals and Human Cells Generated with CRISPR/Cas9 System

    PubMed Central

    Zhu, Xiaoxiao; Xu, Yajie; Yu, Shanshan; Lu, Lu; Ding, Mingqin; Cheng, Jing; Song, Guoxu; Gao, Xing; Yao, Liangming; Fan, Dongdong; Meng, Shu; Zhang, Xuewen; Hu, Shengdi; Tian, Yong

    2014-01-01

    The rapid generation of various species and strains of laboratory animals using CRISPR/Cas9 technology has dramatically accelerated the interrogation of gene function in vivo. So far, the dominant approach for genotyping of genome-modified animals has been the T7E1 endonuclease cleavage assay. Here, we present a polyacrylamide gel electrophoresis-based (PAGE) method to genotype mice harboring different types of indel mutations. We developed 6 strains of genome-modified mice using CRISPR/Cas9 system, and utilized this approach to genotype mice from F0 to F2 generation, which included single and multiplexed genome-modified mice. We also determined the maximal detection sensitivity for detecting mosaic DNA using PAGE-based assay as 0.5%. We further applied PAGE-based genotyping approach to detect CRISPR/Cas9-mediated on- and off-target effect in human 293T and induced pluripotent stem cells (iPSCs). Thus, PAGE-based genotyping approach meets the rapidly increasing demand for genotyping of the fast-growing number of genome-modified animals and human cell lines created using CRISPR/Cas9 system or other nuclease systems such as TALEN or ZFN. PMID:25236476

  17. Novel Variant of Tickborne Encephalitis Virus, Russia

    PubMed Central

    Ternovoi, Vladimir A.; Protopopova, Elena V.; Chausov, Eugene V.; Novikov, Dmitry V.; Leonova, Galina N.; Netesov, Sergey V.

    2007-01-01

    We isolated a novel strain of tickborne encephalitis virus (TBEV), Glubinnoe/2004, from a patient with a fatal case in Russia. We sequenced the strain, whose landmark features included 57 amino acid substitutions and 5 modified cleavage sites. Phylogenetically, Glubinnoe/2004 is a novel variant that belongs to the Eastern type of TBEV. PMID:18258012

  18. Experimental and modeling results of creep fatigue life of Inconel 617 and Haynes 230 at 850 C

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

    Chen, Xiang; Sokolov, Mikhail A; Sham, Sam

    Creep fatigue testing of Ni-based superalloy Inconel 617 and Haynes 230 were conducted in the air at 850 C. Tests were performed with fully reversed axial strain control at a total strain range of 0.5%, 1.0% or 1.5% and hold time at maximum tensile strain for 3, 10 or 30 min. In addition, two creep fatigue life prediction methods, i.e. linear damage summation and frequency-modified tensile hysteresis energy modeling, were evaluated and compared with experimental results. Under all creep fatigue tests, Haynes 230 performed better than Inconel 617. Compared to the low cycle fatigue life, the cycles to failure formore » both materials decreased under creep fatigue test conditions. Longer hold time at maximum tensile strain would cause a further reduction in both material creep fatigue life. The linear damage summation could predict the creep fatigue life of Inconel 617 for limited test conditions, but considerably underestimated the creep fatigue life of Haynes 230. In contrast, frequency-modified tensile hysteresis energy modeling showed promising creep fatigue life prediction results for both materials.« less

  19. [Safety and efficacy of an antirabies vaccine consisting of recombinant vaccinia-rabies virus administered orally to the fox, dog and cat].

    PubMed

    Blancou, J; Artois, M; Brochier, B; Thomas, I; Pastoret, P P; Desmettre, P; Languet, B; Kiény, M P

    1989-01-01

    One of the most promising ways to control rabies in wildlife seems to be the distribution of bait containing an anti-rabies vaccine. So far, the most widely used vaccines were modified live viruses (SAD strain or derivatives). Nevertheless, these strains retain some pathogenicity for non-target species. A novel vaccine was proposed consisting of genetically modified vaccinia virus (strain Copenhagen, thermosensitive ts 26) expressing the foreign glycoprotein G for the rabies virus (strain ERA). Different doses of this recombinant virus were administered orally to 59 foxes (Vulpes vulpes) and their antibodies were titrated before challenge. Foxes (8/8) resisted 1 month after vaccination with 10(7) plaque forming units (PFU), or 4/4 after 18 months. Seroconversion among dogs was 4/4 after vaccination with 10(9,6) PFU and 4/4 among cats after vaccination with 10(8) PFU. These dogs (4/4) and cats (3/4) resisted the challenge 2-3 months after vaccination. This vaccine thus appears to be potent and safe in these species. Its properties are discussed.

  20. Antigenic characterization of a formalin-inactivated poliovirus vaccine derived from live-attenuated Sabin strains.

    PubMed

    Tano, Yoshio; Shimizu, Hiroyuki; Martin, Javier; Nishimura, Yorihiro; Simizu, Bunsiti; Miyamura, Tatsuo

    2007-10-10

    A candidate inactivated poliovirus vaccine derived from live-attenuated Sabin strains (sIPV), which are used in the oral poliovirus vaccine (OPV), was prepared in a large-production scale. The modification of viral antigenic epitopes during the formalin inactivation process was investigated by capture ELISA assays using type-specific and antigenic site-specific monoclonal antibodies (MoAbs). The major antigenic site 1 was modified during the formalin inactivation of Sabin 1. Antigenic sites 1-3 were slightly modified during the formalin inactivation of Sabin 2 strain. Sites 1 and 3 were altered on inactivated Sabin 3 virus. These alterations were different to those shown by wild-type Saukett strain, used in conventional IPV (cIPV). It has been previously reported that type 1 sIPV showed higher immunogenicity to type 1 cIPV whereas types 2 and 3 sIPV induced lower level of immunogenicity than their cIPV counterparts. Our results suggest that the differences in epitope structure after formalin inactivation may account, at least in part, for the observed differences in immunogenicity between Sabin and wild-type inactivated poliovaccines.

  1. Experimental and modeling results of creep-fatigue life of Inconel 617 and Haynes 230 at 850 °C

    NASA Astrophysics Data System (ADS)

    Chen, Xiang; Sokolov, Mikhail A.; Sham, Sam; Erdman, Donald L., III; Busby, Jeremy T.; Mo, Kun; Stubbins, James F.

    2013-01-01

    Creep-fatigue testing of Ni-based superalloy Inconel 617 and Haynes 230 were conducted in the air at 850 °C. Tests were performed with fully reversed axial strain control at a total strain range of 0.5%, 1.0% or 1.5% and hold time at maximum tensile strain for 3, 10 or 30 min. In addition, two creep-fatigue life prediction methods, i.e. linear damage summation and frequency-modified tensile hysteresis energy modeling, were evaluated and compared with experimental results. Under all creep-fatigue tests, Haynes 230 performed better than Inconel 617. Compared to the low cycle fatigue life, the cycles to failure for both materials decreased under creep-fatigue test conditions. Longer hold time at maximum tensile strain would cause a further reduction in both material creep-fatigue life. The linear damage summation could predict the creep-fatigue life of Inconel 617 for limited test conditions, but considerably underestimated the creep-fatigue life of Haynes 230. In contrast, frequency-modified tensile hysteresis energy modeling showed promising creep-fatigue life prediction results for both materials.

  2. Stress concentration investigations using NASTRAN

    NASA Technical Reports Server (NTRS)

    Gillcrist, M. C.; Parnell, L. A.

    1986-01-01

    Parametic investigations are performed using several two dimensional finite element formulations to determine their suitability for use in predicting extremum stresses in marine propellers. Comparisons are made of two NASTRAN elements (CTRIM6 and CTRAIA2) wherein elasticity properties have been modified to yield plane strain results. The accuracy of the elements is investigated by comparing finite element stress predictions with experimentally determined stresses in two classical cases: (1) tension in a flat plate with a circular hole; and (2) a filleted flat bar subjected to in-plane bending. The CTRIA2 element is found to provide good results. The displacement field from a three dimensional finite element model of a representative marine propeller is used as the boundary condition for the two dimensional plane strain investigations of stresses in the propeller blade and fillet. Stress predictions from the three dimensional analysis are compared with those from the two dimensional models. The validity of the plane strain modifications to the NASTRAN element is checked by comparing the modified CTRIA2 element stress predictions with those of the ABAQUS plane strain element, CPE4.

  3. Genomic and antigenic characterization of bovine parainfluenze-3 viruses in the United States including modified live virus vaccine (MLV) strains and field strains from cattle

    USDA-ARS?s Scientific Manuscript database

    This study investigated the genetic and antigenic characterization of parainfluenza-3 virus (PI3V) of cattle. Using molecular tests including real time PCR and viral genome sequencing, PI3V strains could be separated into PI3V types, including PI3V A, PI3V B, and PI3V C. Isolates from cattle with bo...

  4. Characteristics of strain-sensitive photonic crystal cavities in a flexible substrate.

    PubMed

    No, You-Shin; Choi, Jae-Hyuck; Kim, Kyoung-Ho; Park, Hong-Gyu

    2016-11-14

    High-index semiconductor photonic crystal (PhC) cavities in a flexible substrate support strong and tunable optical resonances that can be used for highly sensitive and spatially localized detection of mechanical deformations in physical systems. Here, we report theoretical studies and fundamental understandings of resonant behavior of an optical mode excited in strain-sensitive rod-type PhC cavities consisting of high-index dielectric nanorods embedded in a low-index flexible polymer substrate. Using the three-dimensional finite-difference time-domain simulation method, we calculated two-dimensional transverse-electric-like photonic band diagrams and the three-dimensional dispersion surfaces near the first Γ-point band edge of unidirectionally strained PhCs. A broken rotational symmetry in the PhCs modifies the photonic band structures and results in the asymmetric distributions and different levels of changes in normalized frequencies near the first Γ-point band edge in the reciprocal space, which consequently reveals strain-dependent directional optical losses and selected emission patterns. The calculated electric fields, resonant wavelengths, and quality factors of the band-edge modes in the strained PhCs show an excellent agreement with the results of qualitative analysis of modified dispersion surfaces. Furthermore, polarization-resolved time-averaged Poynting vectors exhibit characteristic dipole-like emission patterns with preferentially selected linear polarizations, originating from the asymmetric band structures in the strained PhCs.

  5. Relation Between Motility, Accelerated Aging and Gene Expression in Selected Drosophila Strains under Hypergravity Conditions

    NASA Astrophysics Data System (ADS)

    Serrano, Paloma; van Loon, Jack J. W. A.; Medina, F. Javier; Herranz, Raúl

    2013-02-01

    Motility and aging in Drosophila have proven to be highly modified under altered gravity conditions (both in space and ground simulation facilities). In order to find out how closely connected they are, five strains with altered geotactic response or survival rates were selected and exposed to an altered gravity environment of 2 g. By analysing the different motile and behavioural patterns and the median survival rates, we show that altered gravity leads to changes in motility, which will have a negative impact on the flies' survival. Previous results show a differential gene expression between sessile samples and adults and confirm that environmentally-conditioned behavioural patterns constrain flies' gene expression and life span. Therefore, hypergravity is considered an environmental stress factor and strains that do not respond to this new environment experience an increment in motility, which is the major cause for the observed increased mortality also under microgravity conditions. The neutral-geotaxis selected strain (strain M) showed the most severe phenotype, unable to respond to variations in the gravitational field. Alternatively, the opposite phenotype was observed in positive-geotaxis and long-life selected flies (strains B and L, respectively), suggesting that these populations are less sensitive to alterations in the gravitational load. We conclude that the behavioural response has a greater contribution to aging than the modified energy consumption in altered gravity environments.

  6. Lactobacillus delbrueckii subsp. lactis (strain CIDCA 133) stimulates murine macrophages infected with Citrobacter rodentium.

    PubMed

    Hugo, Ayelén A; Rolny, Ivanna S; Romanin, David; Pérez, Pablo F

    2017-03-01

    Citrobacter rodentium is a specific murine enteropathogen which causes diarrheal disease characterized by colonic hyperplasia and intestinal inflammation. Recruitment of neutrophils and macrophages constitute a key step to control the infection. Since modulation of the activity of professional phagocytic cells could contribute to improve host´s defences against C. rodentium, we investigated the effect of Lactobacillus delbrueckii subsp. lactis (strain CIDCA 133) on the interaction between murine macrophages (RAW 264.7) and C. rodentium. Phagocytosis, surface molecules and inducible nitric oxide synthase (iNOs) expression were determined by flow cytometry. Reactive oxygen species (ROS) were assessed by fluorescence microscopy. The presence of lactobacilli increased phagocytosis of C. rodentium whereas C. rodentium had no effect on lactobacilli internalization. Survival of internalized C. rodentium diminished when strain CIDCA 133 was present. CD-86, MHCII, iNOs expression and nitrite production were increased when C. rodentium and lactobacilli were present even though strain CIDCA 133 alone had no effect. Strain CIDCA 133 led to a strong induction of ROS activity which was not modified by C. rodentium. Lactobacillus delbrueckii subsp. lactis (strain CIDCA 133) is able to increase the activation of murine macrophages infected with C. rodentium. The sole presence of lactobacilli is enough to modify some stimulation markers (e.g. ROS induction) whereas other markers require the presence of both bacteria; thus, indicating a synergistic effect.

  7. How Does Circadian Rhythm Impact Salt Sensitivity of Blood Pressure in Mice? A Study in Two Close C57Bl/6 Substrains.

    PubMed

    Combe, Roy; Mudgett, John; El Fertak, Lahcen; Champy, Marie-France; Ayme-Dietrich, Estelle; Petit-Demoulière, Benoit; Sorg, Tania; Herault, Yann; Madwed, Jeffrey B; Monassier, Laurent

    2016-01-01

    Mouse transgenesis has provided the unique opportunity to investigate mechanisms underlying sodium kidney reabsorption as well as end organ damage. However, understanding mouse background and the experimental conditions effects on phenotypic readouts of engineered mouse lines such as blood pressure presents a challenge. Despite the ability to generate high sodium and chloride plasma levels during high-salt diet, observed changes in blood pressure are not consistent between wild-type background strains and studies. The present work was designed in an attempt to determine guidelines in the field of salt-induced hypertension by recording continuously blood pressure by telemetry in mice submitted to different sodium and potassium loaded diets and changing experimental conditions in both C57BL/6N and C57BL/6J mice strain (Normal salt vs. Low salt vs. High-salt/normal potassium vs. High salt/low potassium, standard vs. modified light cycle, Non-invasive tail cuff blood pressure vs. telemetry). In this study, we have shown that, despite a strong blood pressure (BP) basal difference between C57BL/6N and C57BL/6J mice, High salt/normal potassium diet increases BP and heart rate during the active phase only (dark period) in the same extent in both strains. On the other hand, while potassium level has no effect on salt-induced hypertension in C57BL/6N mice, high-salt/low potassium diet amplifies the effect of the high-salt challenge only in C57BL/6J mice. Indeed, in this condition, salt-induced hypertension can also be detected during light period even though this BP increase is lower compared to the one occurring during the dark period. Finally, from a methodological perspective, light cycle inversion has no effect on this circadian BP phenotype and tail-cuff method is less sensitive than telemetry to detect BP phenotypes due to salt challenges. Therefore, to carry investigations on salt-induced hypertension in mice, chronic telemetry and studies in the active phase are essential prerequisites.

  8. Impact and Blast Resistance of Sandwich Plates

    NASA Astrophysics Data System (ADS)

    Dvorak, George J.; Bahei-El-Din, Yehia A.; Suvorov, Alexander P.

    Response of conventional and modified sandwich plate designs is examined under static load, impact by a rigid cylindrical or flat indenter, and during and after an exponential pressure impulse lasting for 0.05 ms, at peak pressure of 100 MPa, simulating a nearby explosion. The conventional sandwich design consists of thin outer (loaded side) and inner facesheets made of carbon/epoxy fibrous laminates, separated by a thick layer of structural foam core. In the three modified designs, one or two thin ductile interlayers are inserted between the outer facesheet and the foam core. Materials selected for the interlayers are a hyperelas-tic rate-independent polyurethane;a compression strain and strain rate dependent, elastic-plastic polyurea;and an elastomeric foam. ABAQUS and LS-Dyna software were used in various response simulations. Performance comparisons between the enhanced and conventional designs show that the modified designs provide much better protection against different damage modes under both load regimes. After impact, local facesheet deflection, core compression, and energy release rate of delamination cracks, which may extend on hidden interfaces between facesheet and core, are all reduced. Under blast or impulse loads, reductions have been observed in the extent of core crushing, facesheet delaminations and vibration amplitudes, and in overall deflections. Similar reductions were found in the kinetic energy and in the stored and dissipated strain energy. Although strain rates as high as 10-4/s1 are produced by the blast pressure, peak strains in the interlayers were too low to raise the flow stress in the polyurea to that in the polyurethane, where a possible rate-dependent response was neglected. Therefore, stiff polyurethane or hard rubber interlayers materials should be used for protection of sandwich plate foam cores against both impact and blast-induced damage.

  9. Genetic background contributes to the co-morbidity of anxiety and depression with audiogenic seizure propensity and responses to fluoxetine treatment.

    PubMed

    Sarkisova, Karine Yu; Fedotova, Irina B; Surina, Natalia M; Nikolaev, Georgy M; Perepelkina, Olga V; Kostina, Zoya A; Poletaeva, Inga I

    2017-03-01

    Anxiety and depression are the most frequent comorbidities of different types of convulsive and non-convulsive epilepsies. Increased anxiety and depression-like phenotype have been described in the genetic absence epilepsy models as well as in models of limbic epilepsy and acquired seizure models, suggesting a neurobiological connection. However, whether anxiety and/or depression are comorbid to audiogenic epilepsy remains unclear. The aim of this study was to investigate whether anxiety or depression-like behavior can be found in rat strains with different susceptibility to audiogenic seizures (AS) and whether chronic fluoxetine treatment affects this co-morbidity. Behavior in the elevated plus-maze and the forced swimming test was studied in four strains: Wistar rats non-susceptible to AS; Krushinsky-Molodkina (KM) strain, selectively bred for AS propensity from outbred Wistar rats; and a selection lines bred for maximal AS expression (strain "4") and for a lack of AS (strain "0") from KM×Wistar F2 hybrids. Effects of chronic antidepressant treatment on AS and behavior were also evaluated. Anxiety and depression levels were higher in KM rats (with AS) compared with Wistar rats (without AS), indicating the comorbidity with AS. However, in strains "4" and "0" with contrasting AS expression, but with a genetic background close to KM rats, anxiety and depression were not as divergent as in KMs versus Wistars. Fluoxetine treatment exerted an antidepressant effect in all rat strains irrespective of its effect on AS. Genetic background contributes substantively to the co-morbidity of anxiety and depression with AS propensity. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Omics Analyses of Trichoderma reesei CBS999.97 and QM6a Indicate the Relevance of Female Fertility to Carbohydrate-Active Enzyme and Transporter Levels

    DOE PAGES

    Tisch, Doris; Pomraning, Kyle R.; Collett, James R.; ...

    2017-09-15

    Here, the filamentous fungus Trichoderma reesei is found predominantly in the tropics but also in more temperate regions, such as Europe, and is widely known as a producer of large amounts of plant cell wall-degrading enzymes. We sequenced the genome of the sexually competent isolate CBS999.97, which is phenotypically different from the female sterile strain QM6a but can cross sexually with QM6a. Transcriptome data for growth on cellulose showed that entire carbohydrate-active enzyme (CAZyme) families are consistently differentially regulated between these strains. We evaluated backcrossed strains of both mating types, which acquired female fertility from CBS999.97 but maintained a mostlymore » QM6a genetic background, and we could thereby distinguish between the effects of strain background and female fertility or mating type. We found clear regulatory differences associated with female fertility and female sterility, including regulation of CAZyme and transporter genes. Analysis of carbon source utilization, transcriptomes, and secondary metabolites in these strains revealed that only a few changes in gene regulation are consistently correlated with different mating types. Different strain backgrounds (QM6a versus CBS999.97) resulted in the most significant alterations in the transcriptomes and in carbon source utilization, with decreased growth of CBS999.97 on several amino acids (for example proline or alanine), which further correlated with the downregulation of genes involved in the respective pathways. In combination, our findings support a role of fertility-associated processes in physiology and gene regulation and are of high relevance for the use of sexual crossing in combining the characteristics of two compatible strains or quantitative trait locus (QTL) analysis.« less

  11. Omics Analyses of Trichoderma reesei CBS999.97 and QM6a Indicate the Relevance of Female Fertility to Carbohydrate-Active Enzyme and Transporter Levels

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

    Tisch, Doris; Pomraning, Kyle R.; Collett, James R.

    Here, the filamentous fungus Trichoderma reesei is found predominantly in the tropics but also in more temperate regions, such as Europe, and is widely known as a producer of large amounts of plant cell wall-degrading enzymes. We sequenced the genome of the sexually competent isolate CBS999.97, which is phenotypically different from the female sterile strain QM6a but can cross sexually with QM6a. Transcriptome data for growth on cellulose showed that entire carbohydrate-active enzyme (CAZyme) families are consistently differentially regulated between these strains. We evaluated backcrossed strains of both mating types, which acquired female fertility from CBS999.97 but maintained a mostlymore » QM6a genetic background, and we could thereby distinguish between the effects of strain background and female fertility or mating type. We found clear regulatory differences associated with female fertility and female sterility, including regulation of CAZyme and transporter genes. Analysis of carbon source utilization, transcriptomes, and secondary metabolites in these strains revealed that only a few changes in gene regulation are consistently correlated with different mating types. Different strain backgrounds (QM6a versus CBS999.97) resulted in the most significant alterations in the transcriptomes and in carbon source utilization, with decreased growth of CBS999.97 on several amino acids (for example proline or alanine), which further correlated with the downregulation of genes involved in the respective pathways. In combination, our findings support a role of fertility-associated processes in physiology and gene regulation and are of high relevance for the use of sexual crossing in combining the characteristics of two compatible strains or quantitative trait locus (QTL) analysis.« less

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

    PubMed

    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.

  13. Factors influencing the expression of T15 idiotopes during an antigen-driven response to phosphorylcholine (PC)

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

    Strickland, F.; Cronkhite, R.; Cerny, J.

    1986-03-01

    The idiotypy of the serum antibody response to PC was examined in twelve inbred strains of mice including several Igh- and h-2 congenics. The sera were collected after primary (1/sup 0/), and secondary (2/sup 0/), and tertiary (3/sup 0/) immunizations with either S. pneumoniae R36a (Pn) or PC coupled to keyhole limpet hemocyanine (PC-KLH). Concentrations of PC-binding activity and of four distinct T15 idiotopes (Id) detectable with monoclonal anti-Id AB1-2, MaId5-4, B36-82 and B24-44 were measured in solid-phase radioimmunoassay. These Id are independent clonotypic markers of the anti-PC antibody repertoire. The serum levels of the Id had kinetics distinct frommore » each other and from the total PC-binding activity. For example, the concentration of AB1-2 decreased after 2/sup 0/ and 3/sup 0/ challenge with PC-KLH while the levels of B36-82 and B24-44 remained stable in several strains. Marked differences in Id serum levels were observed among various strains; however, their genetic mapping has been difficult in that an apparent effect of H-2 locus seemed to be modified by the genetic background of the mice. The only unambiguous linkage was the absence of AB1-2 and MaId5-4 in mice bearing Igh/sup j/. The authors data indicate that idiotopically-distinct T15/sup +/ clones are selected during an antigen-driven response in genetically different mice. Mechanisms regulating these patterns are being studied.« less

  14. Heterosis Increases Fertility, Fecundity, and Survival of Laboratory-Produced F1 Hybrid Males of the Malaria Mosquito Anopheles coluzzii

    PubMed Central

    Ekechukwu, Nkiru E.; Baeshen, Rowida; Traorè, Sékou F.; Coulibaly, Mamadou; Diabate, Abdoulaye; Catteruccia, Flaminia; Tripet, Frédéric

    2015-01-01

    The success of vector control strategies aiming to decrease disease transmission via the release of sterile or genetically-modified male mosquitoes critically depends on mating between laboratory-reared males and wild females. Unfortunately, mosquito colonization, laboratory rearing, and genetic manipulations can all negatively affect male competitiveness. Heterosis is commonly used to produce domestic animals with enhanced vigor and homogenous genetic background and could therefore potentially improve the mating performance of mass-reared male mosquitoes. Here, we produced enhanced hybrid males of the malaria mosquito Anopheles coluzzii by crossing two strains colonized >35 and 8 years ago. We compared the amount of sperm and mating plug proteins they transferred to females, as well as their insemination rate, reproductive success and longevity under various experimental conditions. Across experiments, widespread adaptations to laboratory mating were detected in the older strain. In large-group mating experiments, no overall hybrid advantage in insemination rates and the amount of sperm and accessory gland proteins transferred to females was detected. Despite higher sperm activity, hybrid males did not appear more fecund. However, individual-male mating and laboratory-swarm experiments revealed that hybrid males, while inseminating fewer females than older inbred males, were significantly more fertile, producing larger mating plugs and drastically increasing female fecundity. Heterotic males also showed increased longevity. These results validate the use of heterosis for creating hybrid males with improved fitness from long-established inbred laboratory strains. Therefore, this simple approach could facilitate disease control strategies based on male mosquito releases with important ultimate benefits to human health. PMID:26497140

  15. Modified microplate method for rapid and efficient estimation of siderophore produced by bacteria.

    PubMed

    Arora, Naveen Kumar; Verma, Maya

    2017-12-01

    In this study, siderophore production by various bacteria amongst the plant-growth-promoting rhizobacteria was quantified by a rapid and efficient method. In total, 23 siderophore-producing bacterial isolates/strains were taken to estimate their siderophore-producing ability by the standard method (chrome azurol sulphonate assay) as well as 96 well microplate method. Production of siderophore was estimated in percent siderophore unit by both the methods. It was observed that data obtained by both methods correlated positively with each other proving the correctness of microplate method. By the modified microplate method, siderophore production by several bacterial strains can be estimated both qualitatively and quantitatively at one go, saving time, chemicals, making it very less tedious, and also being cheaper in comparison with the method currently in use. The modified microtiter plate method as proposed here makes it far easier to screen the plant-growth-promoting character of plant-associated bacteria.

  16. Achieving high aspect ratio wrinkles by modifying material network stress.

    PubMed

    Chen, Yu-Cheng; Wang, Yan; McCarthy, Thomas J; Crosby, Alfred J

    2017-06-07

    Wrinkle aspect ratio, or the amplitude divided by the wavelength, is hindered by strain localization transitions when an increasing global compressive stress is applied to synthetic material systems. However, many examples from living organisms show extremely high aspect ratios, such as gut villi and flower petals. We use three experimental approaches to demonstrate that these high aspect ratio structures can be achieved by modifying the network stress in the wrinkle substrate. We modify the wrinkle stress and effectively delay the strain localization transition, such as folding, to larger aspect ratios by using a zero-stress initial wavy substrate, creating a secondary network with post-curing, or using chemical stress relaxation materials. A wrinkle aspect ratio as high as 0.85, almost three times higher than common values of synthetic wrinkles, is achieved, and a quantitative framework is presented to provide understanding the different strategies and predictions for future investigations.

  17. A Recombinant Saccharomyces cerevisiae Strain Overproducing Mannoproteins Stabilizes Wine against Protein Haze▿

    PubMed Central

    Gonzalez-Ramos, Daniel; Cebollero, Eduardo; Gonzalez, Ramon

    2008-01-01

    Stabilization against protein haze was one of the first positive properties attributed to yeast mannoproteins in winemaking. In previous work we demonstrated that deletion of KNR4 leads to increased mannoprotein release in laboratory Saccharomyces cerevisiae strains. We have now constructed strains with KNR4 deleted in two different industrial wine yeast backgrounds. This required replacement of two and three alleles of KNR4 for the EC1118 and T73-4 backgrounds, respectively, and the use of three different selection markers for yeast genetic transformation. The actual effect of the genetic modification was dependent on both the genetic background and the culture conditions. The fermentation performance of T73-4 derivatives was clearly impaired, and these derivatives did not contribute to the protein stability of the wine, even though they showed increased mannoprotein release in vitro. In contrast, the EC1118 derivative with both alleles of KNR4 deleted released increased amounts of mannoproteins both in vitro and during wine fermentation assays, and the resulting wines were consistently less susceptible to protein haze. The fermentation performance of this strain was slightly impaired, but only with must with a very high sugar content. These results pave the way for the development of new commercial strains with the potential to improve several mannoprotein-related quality and technological parameters of wine. PMID:18606802

  18. Pyocyanine Biosynthetic Genes in Clinical and Environmental Isolates of Pseudomonas aeruginosa and Detection of Pyocyanine’s Antimicrobial Effects with or without Colloidal Silver Nanoparticles

    PubMed Central

    Nowroozi, Jamileh; Akhavan Sepahi, Abbas; Rashnonejad, Afrooz

    2012-01-01

    Objective: Pyocyanine plays an important role in the pathogenesis of Pseudomonas aeruginosa, (P. aeruginosa) and is known to have inhibitory and bactericidal effects. This study has aimed to detect the phenazine biosynthetic operon (phz ABCDEFG) and two phenazine modifying genes (phzM and phzS) by polymerase chain reaction (PCR) and detection of its possible protein bands by sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE). The antimicrobial effects of pyocyanine alone and mixed with colloidal silver nanoparticles were studied. Materials and Methods: In this descriptive study, clinical and environmental species of P. aeruginosa were isolated by thioglycollate medium culture and cetrimide agar, respectively. The existence of a phenazine biosynthetic operon and two phenazine modifying genes as well as their protein products were confirmed by PCR and SDS-PAGE, respectively. Pyocyanine was extracted with chloroform and its antimicrobial effects against bacteria such as; Escherichia coli (E. coli), P. aeruginosaand Staphylococcus aureus (S. aureus) bacteria and yeast Candida albicans (C. albicans) were tested using well, spot and disk diffusion methods. Results: In this study, 3 out of 48 clinical strains were unable to produce pyocyanine on cetrimide and Mueller Hinton (MH) agar. Two strains did not have phenazine modifying gene bands. Another strain did not have the possible protein band of the phzM gene. Pyocyanine had antimicrobial effects against the microbial strains, which increased in the presence of silver nanoparticles. Conclusion: According to the results of the present study, some P. aeruginosa strains are unable to produce pyocyanine due to the absence of the phzM or phzS genes. Therefore, these genes have an important role in pyocyanine production in P. aeruginosa. Pyocyanine shows synergistic antimicrobial effects in the presence of silver nanoparticles against microbial strains. PMID:23626932

  19. Persistence of Pathogenic and Non-Pathogenic Escherichia coli Strains in Various Tropical Agricultural Soils of India

    PubMed Central

    Naganandhini, S.; Kennedy, Z. John; Uyttendaele, M.; Balachandar, D.

    2015-01-01

    The persistence of Shiga-like toxin producing E. coli (STEC) strains in the agricultural soil creates serious threat to human health through fresh vegetables growing on them. However, the survival of STEC strains in Indian tropical soils is not yet understood thoroughly. Additionally how the survival of STEC strain in soil diverges with non-pathogenic and genetically modified E. coli strains is also not yet assessed. Hence in the present study, the survival pattern of STEC strain (O157-TNAU) was compared with non-pathogenic (MTCC433) and genetically modified (DH5α) strains on different tropical agricultural soils and on a vegetable growing medium, cocopeat under controlled condition. The survival pattern clearly discriminated DH5α from MTCC433 and O157-TNAU, which had shorter life (40 days) than those compared (60 days). Similarly, among the soils assessed, the red laterite and tropical latosol supported longer survival of O157-TNAU and MTCC433 as compared to wetland and black cotton soils. In cocopeat, O157 recorded significantly longer survival than other two strains. The survival data were successfully analyzed using Double-Weibull model and the modeling parameters were correlated with soil physico-chemical and biological properties using principal component analysis (PCA). The PCA of all the three strains revealed that pH, microbial biomass carbon, dehydrogenase activity and available N and P contents of the soil decided the survival of E. coli strains in those soils and cocopeat. The present research work suggests that the survival of O157 differs in tropical Indian soils due to varied physico-chemical and biological properties and the survival is much shorter than those reported in temperate soils. As the survival pattern of non-pathogenic strain, MTCC433 is similar to O157-TNAU in tropical soils, the former can be used as safe model organism for open field studies. PMID:26101887

  20. In vitro susceptibility of Prototheca spp. to gentamicin.

    PubMed Central

    Shahan, T A; Pore, R S

    1991-01-01

    One hundred strains of Prototheca zopfii, Prototheca wickerhamii, Prototheca moriformis, Prototheca stagnora, and Prototheca ulmnea; five strains of Chlorella protothecoides; and two strains of Candida albicans were obtained from a number of different clinical and environmental sources and were tested for their in vitro susceptibility to the antibacterial agent gentamicin. All Prototheca strains were susceptible to gentamicin at concentrations between 0.3 and 0.9 micrograms/ml. A modified macrobroth dilution MIC assay with a colorimeter and a microbroth dilution assay with a 96-well plate reader were the two methods used to determine the MICs. PMID:1804021

  1. Effects of misalignment on mechanical behavior of metals in creep

    NASA Technical Reports Server (NTRS)

    Wu, H. C.

    1981-01-01

    Creep tests were conducted by means of a closed loop servocontrolled materials test system. The strain history prior to creep is carefully monitored. Tests were performed for aluminum alloy 6061-O at 150 C and were monitored by a PDP 11/04 minicomputer at a preset constant plastic strain rate prehistory. The results show that the plastic strain rate prior to creep plays a significant role in creep behavior. The endochronic theory of viscoplasticity was applied to describe the observed creep curves. Intrinsic time and strain rate sensitivity function concepts are employed and modified according to the present observation.

  2. Process-driven bacterial community dynamics are key to cured meat colour formation by coagulase-negative staphylococci via nitrate reductase or nitric oxide synthase activities.

    PubMed

    Sánchez Mainar, María; Leroy, Frédéric

    2015-11-06

    The cured colour of European raw fermented meats is usually achieved by nitrate-into-nitrite reduction by coagulase-negative staphylococci (CNS), subsequently generating nitric oxide to form the relatively stable nitrosomyoglobin pigment. The present study aimed at comparing this classical curing procedure, based on nitrate reductase activity, with a potential alternative colour formation mechanism, based on nitric oxide synthase (NOS) activity, under different acidification profiles. To this end, meat models with and without added nitrate were fermented with cultures of an acidifying strain (Lactobacillus sakei CTC 494) and either a nitrate-reducing Staphylococcus carnosus strain or a rare NOS-positive CNS strain (Staphylococcus haemolyticus G110), or by relying on the background microbiota. Satisfactory colour was obtained in the models prepared with added nitrate and S. carnosus. In the presence of nitrate but absence of added CNS, however, cured colour was only obtained when L. sakei CTC 494 was also omitted. This was ascribed to the pH dependency of the emerging CNS background microbiota, selecting for nitrate-reducing Staphylococcus equorum strains at mild acidification conditions but for Staphylococcus saprophyticus strains with poor colour formation capability when the pH decrease was more rapid. This reliance of colour formation on the composition of the background microbiota was further explored by a side experiment, demonstrating the heterogeneity in nitrate reduction of a set of 88 CNS strains from different species. Finally, in all batches prepared with S. haemolyticus G110, colour generation failed as the strain was systematically outcompeted by the background microbiota, even when imposing milder acidification profiles. Thus, when aiming at colour formation through CNS metabolism, technological processing can severely interfere with the composition and functionality of the meat-associated CNS communities, for both nitrate reductase and NOS activities. Several major bottlenecks, among which the rareness of phenotypic NOS activity in meat-compatible CNS, need to be considered, which is seriously questioning the relevance of this pathway in fermented meats. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. New Lager Brewery Strains Obtained by Crossing Techniques Using Cachaça (Brazilian Spirit) Yeasts

    PubMed Central

    Figueiredo, Bruna Inez Carvalho; Saraiva, Margarete Alice Fontes; de Souza Pimenta, Paloma Patrick; de Souza Testasicca, Miriam Conceição; Sampaio, Geraldo Magela Santos; da Cunha, Aureliano Claret; Afonso, Luis Carlos Crocco; Vieira de Queiroz, Marisa; de Miranda Castro, Ieso

    2017-01-01

    ABSTRACT The development of hybrids has been an effective approach to generate novel yeast strains with optimal technological profile for use in beer production. This study describes the generation of a new yeast strain for lager beer production by direct mating between two Saccharomyces cerevisiae strains isolated from cachaça distilleries: one that was strongly flocculent, and the other with higher production of acetate esters. The first step in this procedure was to analyze the sporulation ability and reproductive cycle of strains belonging to a specific collection of yeasts isolated from cachaça fermentation vats. Most strains showed high rates of sporulation, spore viability, and homothallic behavior. In order to obtain new yeast strains with desirable properties useful for lager beer production, we compare haploid-to-haploid and diploid-to-diploid mating procedures. Moreover, an assessment of parental phenotype traits showed that the segregant diploid C2-1d generated from a diploid-to-diploid mating experiment showed good fermentation performance at low temperature, high flocculation capacity, and desirable production of acetate esters that was significantly better than that of one type lager strain. Therefore, strain C2-1d might be an important candidate for the production of lager beer, with distinct fruit traces and originating using a non-genetically modified organism (GMO) approach. IMPORTANCE Recent work has suggested the utilization of hybridization techniques for the generation of novel non-genetically modified brewing yeast strains with combined properties not commonly found in a unique yeast strain. We have observed remarkable traits, especially low temperature tolerance, maltotriose utilization, flocculation ability, and production of volatile aroma compounds, among a collection of Saccharomyces cerevisiae strains isolated from cachaça distilleries, which allow their utilization in the production of beer. The significance of our research is in the use of breeding/hybridization techniques to generate yeast strains that would be appropriate for producing new lager beers by exploring the capacity of cachaça yeast strains to flocculate and to ferment maltose at low temperature, with the concomitant production of flavoring compounds. PMID:28778887

  4. L-malate production by metabolically engineered escherichia coli

    DOEpatents

    Zhang, Xueli; Wang, Xuan; Shanmugam, Keelnatham T.; Ingram, Lonnie O'Neal

    2015-11-17

    A process for the production of malic acid in commercially significant quantities from the carbon compounds by genetically modified bacterial strains (GMBS; also referred to as biocatalysts or genetically modified microorganisms) is disclosed. Microorganisms suitable for the production of malic acid can be cultured in one or two-step processes as disclosed herein.

  5. Aggregate structure and effect of phthalic anhydride modified soy protein on the mechanical properties of styrene-butadiene copolymer

    USDA-ARS?s Scientific Manuscript database

    The aggregate structure of phthalic anhydride (PA) modified soy protein isolate (SPI) was investigated by estimating its fractal dimension from the equilibrated dynamic strain sweep experiments. The estimated fractal dimensions of the filler aggregates were less than 2, indicating that these partic...

  6. Biological and molecular characterizations of Toxoplasma gondii strains obtained from Southern sea otters (Enhydra lutris nereis)

    USGS Publications Warehouse

    Cole, Rebecca A.; Lindsay, D.S.; Howe, D.K.; Roderick, Constance L.; Dubey, J.P.; Thomas, N.J.; Baeten, L.A.

    2000-01-01

    Toxoplasma gondii was isolated from brain or heart tissue from 15 southern sea otters (Enhydra lutris nereis) in cell cultures. These strains were used to infect mice that developed antibodies to T. gondii as detected in the modified direct agglutination test and had T. gondii tissue cysts in their brains at necropsy. Mouse brains containing tissue cysts from 4 of the strains were fed to 4 cats. Two of the cats excreted T. gondii oocysts in their feces that were infectious for mice. Molecular analyses of 13 strains indicated that they were all type II strains, but that they were genetically distinct from one another.

  7. Study of strain boundary conditions and GaAs buffer sizes in InGaAs quantum dots

    NASA Technical Reports Server (NTRS)

    Oyafuso, F.; Klimeck, G.; Boykin, T. B.; Bowen, R. C.; Allmen, P. von

    2003-01-01

    NEMO 3-D has been developed for the simulation of electronic structure in self-assembled InGaAs quantum dots on GaAs substrates. Typical self-assembled quantum dots in that material system contain about 0.5 to 1 million atoms. Effects of strain by the surrounding GaAs buffer modify the electronic structure inside the quantum dot significantly and a large GaAs buffer must be included in the strain and electronic structure.

  8. L-arabinose fermenting yeast

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

    Zhang, Min; Singh, Arjun; Suominen, Pirkko

    An L-arabinose utilizing yeast strain is provided for the production of ethanol by introducing and expressing bacterial araA, araB and araD genes. L-arabinose transporters are also introduced into the yeast to enhance the uptake of arabinose. The yeast carries additional genomic mutations enabling it to consume L-arabinose, even as the only carbon source, and to produce ethanol. A yeast strain engineered to metabolize arabinose through a novel pathway is also disclosed. Methods of producing ethanol include utilizing these modified yeast strains.

  9. L-arabinose fermenting yeast

    DOEpatents

    Zhang, Min; Singh, Arjun; Suominen, Pirkko; Knoshaug, Eric; Franden, Mary Ann; Jarvis, Eric

    2014-09-23

    An L-arabinose utilizing yeast strain is provided for the production of ethanol by introducing and expressing bacterial araA, araB and araD genes. L-arabinose transporters are also introduced into the yeast to enhance the uptake of arabinose. The yeast carries additional genomic mutations enabling it to consume L-arabinose, even as the only carbon source, and to produce ethanol. A yeast strain engineered to metabolize arabinose through a novel pathway is also disclosed. Methods of producing ethanol include utilizing these modified yeast strains.

  10. Modeling and Characterization of PMMA for High Strain-Rate and Finite Deformations (Postprint)

    DTIC Science & Technology

    2010-05-01

    List of parameters for the modified MuUiken- model for PMMA . Von Mises [MPa] ^AJ3 V 00 ^ Aa ^Afi CR ha hp Value 3386 1748 0.35 298 1979...AFRL-RW-EG-TP-2010-073 Modeling and Characterization of PMMA for High Strain-Rate and Finite Deformations (Postprint) Eric B. Herbold Jennifer L...SUBTITLE Modeling and Characterization of PMMA for High Strain-Rate and Finite Deformations (Postprint) 5a. CONTRACT NUMBER 5b. GRANT NUMBER

  11. Fiber Optic Rosette Strain Gauge Development and Application on a Large-Scale Composite Structure

    NASA Technical Reports Server (NTRS)

    Moore, Jason P.; Przekop, Adam; Juarez, Peter D.; Roth, Mark C.

    2015-01-01

    A detailed description of the construction, application, and measurement of 196 FO rosette strain gauges that measured multi-axis strain across the outside upper surface of the forward bulkhead component of a multibay composite fuselage test article is presented. A background of the FO strain gauge and the FO measurement system as utilized in this application is given and results for the higher load cases of the testing sequence are shown.

  12. Evaluation of a Recombinant Escherichia coli Strain that Uses the Sarin Simulant Isopropylmethylphosphonic Acid (IMPA) as a Sole Carbon and Phosphate Source

    DTIC Science & Technology

    2016-04-01

    phosphate use by these recombinant strains was evaluated because carbon use by these strains is still undergoing optimization by LBNL. The E . coli ...plasmids, had successful growth when transformed into a different E . coli background, which correlated with IMPA degradation. Ultimately, the...transformed E . coli strains, optimized at ECBC, were able to grow using IMPA as the phosphate source. 15. SUBJECT TERMS Acetylcholinesterase (AChE

  13. Optochin Resistance among Streptococcus pneumoniae Strains Colonizing Healthy Children in Portugal▿

    PubMed Central

    Nunes, Sónia; Sá-Leão, Raquel; de Lencastre, Hermínia

    2008-01-01

    Two percent of 1,973 pneumococcus strains isolated from carriers since 2001 in Portugal were found to be optochin resistant. These strains belonged to eight serotypes (and some were nontypeable), and they had diverse genetic backgrounds. Novel optochin-resistant lineages were detected over time, suggesting that there was a continuous, although sporadic, emergence of optochin resistance. PMID:18032618

  14. Genetic factors modulate the impact of pubertal androgen excess on insulin sensitivity and fertility.

    PubMed

    Dowling, Abigail R; Nedorezov, Laura B; Qiu, Xiaoliang; Marino, Joseph S; Hill, Jennifer W

    2013-01-01

    Polycystic ovary syndrome (PCOS) is the most common endocrine disorder of reproductive age women. The syndrome is caused by a combination of environmental influences and genetic predisposition. Despite extensive efforts, the heritable factors contributing to PCOS development are not fully understood. The objective of this study was to test the hypothesis that genetic background contributes to the development of a PCOS-like reproductive and metabolic phenotype in mice exposed to excess DHEA during the pubertal transition. We tested whether the PCOS phenotype would be more pronounced on the diabetes-prone C57BL/6 background than the previously used strain, BALB/cByJ. In addition, we examined strain-dependent upregulation of the expression of ovarian and extra-ovarian candidate genes implicated in human PCOS, genes containing known strain variants, and genes involved with steroidogenesis or insulin sensitivity. These studies show that there are significant strain-related differences in metabolic response to excess androgen exposure during puberty. Additionally, our results suggest the C57BL/6J strain provides a more robust and uniform experimental platform for PCOS research than the BALB/cByJ strain.

  15. Role of scaffold network in controlling strain and functionalities of nanocomposite films

    DOE PAGES

    Chen, Aiping; Hu, Jia -Mian; Lu, Ping; ...

    2016-06-10

    One novel approach to manipulating functionalities in correlated complex oxides is strain. However, significant epitaxial strain can only be achieved in ultrathin layers. We show that, under direct lattice matching framework, large and uniform vertical strain up to 2% can be achieved to significantly modify the magnetic anisotropy, magnetism, and magnetotransport properties in heteroepitaxial nanoscaffold films, over a few hundred nanometers in thickness. Comprehensive designing principles of large vertical strain have been proposed. Phase-field simulations not only reveal the strain distribution but also suggest that the ultimate strain is related to the vertical interfacial area and interfacial dislocation density. Moreover,more » by changing the nanoscaffold density and dimension, the strain and the magnetic properties can be tuned. The established correlation among the vertical interface—strain—properties in nanoscaffold films can consequently be used to tune other functionalities in a broad range of complex oxide films far beyond critical thickness.« less

  16. Role of scaffold network in controlling strain and functionalities of nanocomposite films

    PubMed Central

    Chen, Aiping; Hu, Jia-Mian; Lu, Ping; Yang, Tiannan; Zhang, Wenrui; Li, Leigang; Ahmed, Towfiq; Enriquez, Erik; Weigand, Marcus; Su, Qing; Wang, Haiyan; Zhu, Jian-Xin; MacManus-Driscoll, Judith L.; Chen, Long-Qing; Yarotski, Dmitry; Jia, Quanxi

    2016-01-01

    Strain is a novel approach to manipulating functionalities in correlated complex oxides. However, significant epitaxial strain can only be achieved in ultrathin layers. We show that, under direct lattice matching framework, large and uniform vertical strain up to 2% can be achieved to significantly modify the magnetic anisotropy, magnetism, and magnetotransport properties in heteroepitaxial nanoscaffold films, over a few hundred nanometers in thickness. Comprehensive designing principles of large vertical strain have been proposed. Phase-field simulations not only reveal the strain distribution but also suggest that the ultimate strain is related to the vertical interfacial area and interfacial dislocation density. By changing the nanoscaffold density and dimension, the strain and the magnetic properties can be tuned. The established correlation among the vertical interface—strain—properties in nanoscaffold films can consequently be used to tune other functionalities in a broad range of complex oxide films far beyond critical thickness. PMID:27386578

  17. Stimulation of d- and l-lactate dehydrogenases transcriptional levels in presence of diammonium hydrogen phosphate resulting to enhanced lactic acid production by Lactobacillus strain.

    PubMed

    Singhvi, Mamata; Zendo, Takeshi; Iida, Hiroshi; Gokhale, Digambar; Sonomoto, Kenji

    2017-12-01

    The present study revealed the effect of nitrogen sources on lactic acid production and stimulation of d- and l-lactate dehydrogenases (LDH) of parent Lactobacillus lactis NCIM 2368 and its mutant RM2-24 generated after UV mutagenesis. Both the parent and mutant strains were evaluated for d-lactic acid production in control and modified media. The modified media did not show remarkable effect on lactic acid production in case of parent whereas mutant exhibited significant enhancement in d-lactic acid production along with the appearance of l-lactic acid in the broth. Both LDH activities and specific activities were found to be higher in mutant than the parent strain. These results suggested that the diammonium hydrogen phosphate in modified media triggered the expression of LDH genes leading to enhanced lactic acid production. This observation has been proved by studying the expression levels of d- and l-LDH genes of parent and mutant in control and modified media using quantitative RT-PCR technique. In case of mutant, the transcriptional levels of d-LDH and l-LDH increased ∼17 fold and ∼1.38 fold respectively in modified medium compared to the values obtained with control medium. In case of parent, no significant change in transcriptional levels of d- and l-LDH was found when the cells were grown in either control medium or modified medium. This study suggested that the mutant, RM2-24 has l-LDH gene which is expressed in presence of (NH 4 ) 2 HPO 4 resulting in l-lactic acid production. Co-production of l-lactic acid in d-lactic acid fermentation may be detrimental in the PLA production. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  18. Heterologous Packaging Signals on Segment 4, but Not Segment 6 or Segment 8, Limit Influenza A Virus Reassortment.

    PubMed

    White, Maria C; Steel, John; Lowen, Anice C

    2017-06-01

    Influenza A virus (IAV) RNA packaging signals serve to direct the incorporation of IAV gene segments into virus particles, and this process is thought to be mediated by segment-segment interactions. These packaging signals are segment and strain specific, and as such, they have the potential to impact reassortment outcomes between different IAV strains. Our study aimed to quantify the impact of packaging signal mismatch on IAV reassortment using the human seasonal influenza A/Panama/2007/99 (H3N2) and pandemic influenza A/Netherlands/602/2009 (H1N1) viruses. Focusing on the three most divergent segments, we constructed pairs of viruses that encoded identical proteins but differed in the packaging signal regions on a single segment. We then evaluated the frequency with which segments carrying homologous versus heterologous packaging signals were incorporated into reassortant progeny viruses. We found that, when segment 4 (HA) of coinfecting parental viruses was modified, there was a significant preference for the segment containing matched packaging signals relative to the background of the virus. This preference was apparent even when the homologous HA constituted a minority of the HA segment population available in the cell for packaging. Conversely, when segment 6 (NA) or segment 8 (NS) carried modified packaging signals, there was no significant preference for homologous packaging signals. These data suggest that movement of NA and NS segments between the human H3N2 and H1N1 lineages is unlikely to be restricted by packaging signal mismatch, while movement of the HA segment would be more constrained. Our results indicate that the importance of packaging signals in IAV reassortment is segment dependent. IMPORTANCE Influenza A viruses (IAVs) can exchange genes through reassortment. This process contributes to both the highly diverse population of IAVs found in nature and the formation of novel epidemic and pandemic IAV strains. Our study sought to determine the extent to which IAV packaging signal divergence impacts reassortment between seasonal IAVs. Our knowledge in this area is lacking, and insight into the factors that influence IAV reassortment will inform and strengthen ongoing public health efforts to anticipate the emergence of new viruses. We found that the packaging signals on the HA segment, but not the NA or NS segments, restricted IAV reassortment. Thus, the packaging signals of the HA segment could be an important factor in determining the likelihood that two IAV strains of public health interest will undergo reassortment. Copyright © 2017 American Society for Microbiology.

  19. Heterologous Packaging Signals on Segment 4, but Not Segment 6 or Segment 8, Limit Influenza A Virus Reassortment

    PubMed Central

    White, Maria C.; Steel, John

    2017-01-01

    ABSTRACT Influenza A virus (IAV) RNA packaging signals serve to direct the incorporation of IAV gene segments into virus particles, and this process is thought to be mediated by segment-segment interactions. These packaging signals are segment and strain specific, and as such, they have the potential to impact reassortment outcomes between different IAV strains. Our study aimed to quantify the impact of packaging signal mismatch on IAV reassortment using the human seasonal influenza A/Panama/2007/99 (H3N2) and pandemic influenza A/Netherlands/602/2009 (H1N1) viruses. Focusing on the three most divergent segments, we constructed pairs of viruses that encoded identical proteins but differed in the packaging signal regions on a single segment. We then evaluated the frequency with which segments carrying homologous versus heterologous packaging signals were incorporated into reassortant progeny viruses. We found that, when segment 4 (HA) of coinfecting parental viruses was modified, there was a significant preference for the segment containing matched packaging signals relative to the background of the virus. This preference was apparent even when the homologous HA constituted a minority of the HA segment population available in the cell for packaging. Conversely, when segment 6 (NA) or segment 8 (NS) carried modified packaging signals, there was no significant preference for homologous packaging signals. These data suggest that movement of NA and NS segments between the human H3N2 and H1N1 lineages is unlikely to be restricted by packaging signal mismatch, while movement of the HA segment would be more constrained. Our results indicate that the importance of packaging signals in IAV reassortment is segment dependent. IMPORTANCE Influenza A viruses (IAVs) can exchange genes through reassortment. This process contributes to both the highly diverse population of IAVs found in nature and the formation of novel epidemic and pandemic IAV strains. Our study sought to determine the extent to which IAV packaging signal divergence impacts reassortment between seasonal IAVs. Our knowledge in this area is lacking, and insight into the factors that influence IAV reassortment will inform and strengthen ongoing public health efforts to anticipate the emergence of new viruses. We found that the packaging signals on the HA segment, but not the NA or NS segments, restricted IAV reassortment. Thus, the packaging signals of the HA segment could be an important factor in determining the likelihood that two IAV strains of public health interest will undergo reassortment. PMID:28331085

  20. A Modified Johnson-Cook Model for Sheet Metal Forming at Elevated Temperatures and Its Application for Cooled Stress-Strain Curve and Spring-Back Prediction

    NASA Astrophysics Data System (ADS)

    Duc-Toan, Nguyen; Tien-Long, Banh; Young-Suk, Kim; Dong-Won, Jung

    2011-08-01

    In this study, a modified Johnson-Cook (J-C) model and an innovated method to determine (J-C) material parameters are proposed to predict more correctly stress-strain curve for tensile tests in elevated temperatures. A MATLAB tool is used to determine material parameters by fitting a curve to follow Ludwick's hardening law at various elevated temperatures. Those hardening law parameters are then utilized to determine modified (J-C) model material parameters. The modified (J-C) model shows the better prediction compared to the conventional one. As the first verification, an FEM tensile test simulation based on the isotropic hardening model for boron sheet steel at elevated temperatures was carried out via a user-material subroutine, using an explicit finite element code, and compared with the measurements. The temperature decrease of all elements due to the air cooling process was then calculated when considering the modified (J-C) model and coded to VUMAT subroutine for tensile test simulation of cooling process. The modified (J-C) model showed the good agreement between the simulation results and the corresponding experiments. The second investigation was applied for V-bending spring-back prediction of magnesium alloy sheets at elevated temperatures. Here, the combination of proposed J-C model with modified hardening law considering the unusual plastic behaviour for magnesium alloy sheet was adopted for FEM simulation of V-bending spring-back prediction and shown the good comparability with corresponding experiments.

  1. Altering thermal transport by strained-layer epitaxy

    NASA Astrophysics Data System (ADS)

    Majdi, Tahereh; Pal, Souvik; Hafreager, Anders; Murad, Sohail; Sahu, Rakesh P.; Puri, Ishwar K.

    2018-05-01

    Since strain changes the interatomic spacing of matter and alters electron and phonon dispersion, an applied strain can modify the thermal conductivity k of a material. We show how the strain induced by heteroepitaxy is a passive mechanism to change k in a thin film. Molecular dynamics simulations of the deposition and epitaxial growth of ZnTe thin films provide insights into the role of interfacial strain in the conductivity of a deposited film. ZnTe films grow strain-free on lattice-matched ZnTe substrates, but similar thin films grown on a lattice-mismatched CdTe substrate exhibit ˜6% biaxial in-plane tensile strain and ˜7% uniaxial out-of-plane compressive strain. In the T = 700 K-1100 K temperature range, the conductivities of strained ZnTe layers decrease to ˜60% of their unstrained values. The resulting understanding of dk/dT shows that strain engineering can be used to alter the performance of a thermal rectifier and also provides a framework for enhancing thermoelectric devices.

  2. Real-time reverse transcription polymerase chain reaction method for detection of Canine distemper virus modified live vaccine shedding for differentiation from infection with wild-type strains.

    PubMed

    Wilkes, Rebecca P; Sanchez, Elena; Riley, Matthew C; Kennedy, Melissa A

    2014-01-01

    Canine distemper virus (CDV) remains a common cause of infectious disease in dogs, particularly in high-density housing situations such as shelters. Vaccination of all dogs against CDV is recommended at the time of admission to animal shelters and many use a modified live virus (MLV) vaccine. From a diagnostic standpoint for dogs with suspected CDV infection, this is problematic because highly sensitive diagnostic real-time reverse transcription polymerase chain reaction (RT-PCR) tests are able to detect MLV virus in clinical samples. Real-time PCR can be used to quantitate amount of virus shedding and can differentiate vaccine strains from wild-type strains when shedding is high. However, differentiation by quantitation is not possible in vaccinated animals during acute infection, when shedding is low and could be mistaken for low level vaccine virus shedding. While there are gel-based RT-PCR assays for differentiation of vaccine strains from field strains based on sequence differences, the sensitivity of these assays is unable to match that of the real-time RT-PCR assay currently used in the authors' laboratory. Therefore, a real-time RT-PCR assay was developed that detects CDV MLV vaccine strains and distinguishes them from wild-type strains based on nucleotide sequence differences, rather than the amount of viral RNA in the sample. The test is highly sensitive, with detection of as few as 5 virus genomic copies (corresponding to 10(-1) TCID(50)). Sequencing of the DNA real-time products also allows phylogenetic differentiation of the wild-type strains. This test will aid diagnosis during outbreaks of CDV in recently vaccinated animals.

  3. Xylitol production by genetically modified industrial strain of Saccharomyces cerevisiae using glycerol as co-substrate.

    PubMed

    Kogje, Anushree B; Ghosalkar, Anand

    2017-06-01

    Xylitol is commercially used in chewing gum and dental care products as a low calorie sweetener having medicinal properties. Industrial yeast strain of S. cerevisiae was genetically modified to overexpress an endogenous aldose reductase gene GRE3 and a xylose transporter gene SUT1 for the production of xylitol. The recombinant strain (XP-RTK) carried the expression cassettes of both the genes and the G418 resistance marker cassette KanMX integrated into the genome of S. cerevisiae. Short segments from the 5' and 3' delta regions of the Ty1 retrotransposons were used as homology regions for integration of the cassettes. Xylitol production by the industrial recombinant strain was evaluated using hemicellulosic hydrolysate of the corn cob with glucose as the cosubstrate. The recombinant strain XP-RTK showed significantly higher xylitol productivity (212 mg L -1  h -1 ) over the control strain XP (81 mg L -1  h -1 ). Glucose was successfully replaced by glycerol as a co-substrate for xylitol production by S. cerevisiae. Strain XP-RTK showed the highest xylitol productivity of 318.6 mg L -1  h -1 and titre of 47 g L -1 of xylitol at 12 g L -1 initial DCW using glycerol as cosubstrate. The amount of glycerol consumed per amount of xylitol produced (0.47 mol mol -1 ) was significantly lower than glucose (23.7 mol mol -1 ). Fermentation strategies such as cell recycle and use of the industrial nitrogen sources were demonstrated using hemicellulosic hydrolysate for xylitol production.

  4. Constitutive Modeling of High-Temperature Flow Behavior of an Nb Micro-alloyed Hot Stamping Steel

    NASA Astrophysics Data System (ADS)

    Zhang, Shiqi; Feng, Ding; Huang, Yunhua; Wei, Shizhong; Mohrbacher, Hardy; Zhang, Yue

    2016-03-01

    The thermal deformation behavior and constitutive models of an Nb micro-alloyed 22MnB5 steel were investigated by conducting isothermal uniaxial tensile tests at the temperature range of 873-1223 K with strain rates of 0.1-10 s-1. The results indicated that the investigated steel showed typical work hardening and dynamic recovery behavior during hot deformation, and the flow stress decreased with a decrease in strain rate and/or an increase in temperature. On the basis of the experimental data, the modified Johnson-Cook (modified JC), modified Norton-Hoff (modified NH), and Arrhenius-type (AT) constitutive models were established for the subject steel. However, the flow stress values predicted by these three models revealed some remarkable deviations from the experimental values for certain experimental conditions. Therefore, a new combined modified Norton-Hoff and Arrhenius-type constitutive model (combined modified NH-AT model), which accurately reflected both the work hardening and dynamic recovery behavior of the subject steel, was developed by introducing the modified parameter k ɛ. Furthermore, the accuracy of these constitutive models was assessed by the correlation coefficient, the average absolute relative error, and the root mean square error, which indicated that the flow stress values computed by the combined modified NH-AT model were highly consistent with the experimental values (R = 0.998, AARE = 1.63%, RMSE = 3.85 MPa). The result confirmed that the combined modified NH-AT model was suitable for the studied Nb micro-alloyed hot stamping steel. Additionally, the practicability of the new model was also verified using finite element simulations in ANSYS/LS-DYNA, and the results confirmed that the new model was practical and highly accurate.

  5. Evidence for the presence of restriction/modification systems in Lactobacillus delbrueckii.

    PubMed

    Suárez, Viviana; Zago, Miriam; Giraffa, Giorgio; Reinheimer, Jorge; Quiberoni, Andrea

    2009-11-01

    The bacteriophages Cb1/204 and Cb1/342 were obtained by induction from the commercial strain Lactobacillus delbrueckii subsp. lactis Cb1, and propagated on Lactobacillus delbrueckii subsp. lactis 204 (Lb.l 204) and Lactobacillus delbrueckii subsp. bulgaricus 342 (Lb.b 342), respectively. By cross sensitivity, it was possible to detect a delay in the lysis of Lb.l 204 with Cb1/342 phage, while the adsorption rate was high (99.5%). Modified and unmodified phages were isolated using phage Cb1/342 and strain Lb.l 204. The EOP (Efficiency of Plaquing) values for the four phages (Cb1/204, Cb1/342, Cb1/342modified and Cb1/342unmodified) suggested that an R/M system modified the original temperate phage, and the BglII-DNA restriction patterns of these phages might point out the presence of a Type II R/M system. Also, the existence of a Type I R/M system was demonstrated by PCR and nucleotide sequence, being the percentages of alignment homology with Type I R/M systems reported previously higher than 95%. In this study it was possible to demonstrate that the native phage resistant mechanisms and the occurrence of prophages in commercial host strains, contribute strongly to diversify the phage population in a factory environment.

  6. Minimizing use of fish meal in sunshine bass diets using standard and new varieties of non-genetically modified soybeans

    USDA-ARS?s Scientific Manuscript database

    Improved plant ingredients are needed to support sustainable culture of carnivorous fish, such as hybrid striped bass (HSB). We are evaluating meals made from new strains of non-genetically-modified soybeans (non-GMO) with high protein and reduced anti-nutritional factors (ANFs) on HSB nutrient dige...

  7. Preinfection of citrus by RB strains of Citrus tristeza virus (CTV) negatively affected expression of a modified T36 CTV vector

    USDA-ARS?s Scientific Manuscript database

    Genetically modified T36 Citrus tristeza virus (T36-mCTV) is showing promise in Florida to mitigate huanglongbing (HLB) by expressing antimicrobial peptides and RNAi against the presumed pathogen, “Candidatus Liberibacter asiaticus” (CLas), and its vector, the Asian citrus psyllid (ACP). To this end...

  8. Vaccination with a modified-live bovine viral diarrhea virus (BVDV) type 1a vaccine completely protected calves against challenge with BVDV type 1b strains.

    PubMed

    Xue, Wenzhi; Mattick, Debra; Smith, Linda; Umbaugh, Jerry; Trigo, Emilio

    2010-12-10

    Vaccination plays a significant role in the control of bovine viral diarrhea virus (BVDV) infection and spread. Recent studies revealed that type 1b is the predominant BVDV type 1 subgenotype, representing more than 75% of field isolates of BVDV-1. However, nearly all current, commercially available BVDV type 1 vaccines contain BVDV-1a strains. Previous studies have indicated that anti-BVDV sera, induced by BVDV-1a viruses, show less neutralization activity to BVDV-1b isolates than type 1a. Therefore, it is critically important to evaluate BVDV-1a vaccines in their ability to prevent BVDV-1b infection in calves. In current studies, calves were vaccinated subcutaneously, intradermally or intranasally with a single dose of a multivalent, modified-live viral vaccine containing a BVDV-1a strain, and were challenged with differing BVDV-1b strains to determine the efficacy and duration of immunity of the vaccine against these heterologous virus strains. Vaccinated calves, in all administration routes, were protected from respiratory disease caused by the BVDV-1b viruses, as indicated by significantly fewer clinical signs, lower rectal temperatures, reduced viral shedding and greater white blood cell counts than non-vaccinated control animals. The BVDV-1a vaccine elicited efficacious protection in calves against each BVDV-1b challenge strain, with a duration of immunity of at least 6 months. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Impacts of Pantoea agglomerans strain and cation-modified clay minerals on the adsorption and biodegradation of phenanthrene.

    PubMed

    Tao, Kelin; Zhao, Song; Gao, Pin; Wang, Lijin; Jia, Hanzhong

    2018-06-06

    Interactions between microorganisms and minerals have the potential contribution to remove polycyclic aromatic hydrocarbons (PAHs) in model systems. In this study, phenanthrene (PHE) was used as a probe molecule to explore the potential adsorption and biotransformation processes in the presence of microorganisms and various reference clays, such as montmorillonite (M), kaolinite (K), and pyrophyllite (P). Equilibrium adsorption experiments and scanning electron microscopy (SEM) technique were used to investigate the sorption of Pantoea agglomerans strains on clay minerals saturated with cations (Na + and Fe 3+ ). The adsorption isotherms of PHE and Pantoea agglomerans strains on cation-modified clay minerals fitted to Langmuir equation, and their adsorbed amounts both followed the sequence: montmorillonite > kaolinite > pyrophyllite. For six types of cation-modified minerals, the behavior of PHE adsorbed and Pantoea agglomerans adhered onto mentioned minerals was in the order of Na(I)-M > Fe(Ⅲ)-M, Na(I)-K > Fe(Ⅲ)-K and Fe(Ⅲ)-P > Na(I)-P, respectively. The biodegradation results showed that cation-modified clay minerals could enhance the biodegradation of PHE, ascribing to their large specific surface area, and cation exchange capability, as well as the difference in zeta potential between minerals and Pantoea agglomerans strains. Comparison of biodegradation rates displayed that PHE was degraded the highest in the presence of Na-M (93.285%). In addition, the obtained results suggested that the adhesion of bacteria onto cation-exchanged clay minerals was beneficial to the biodegradation of PHE. Anthracen-9-ylmethanol and 3,4-dimethyl-2-(3-methylbutanoyl)benzoic acid were detected as the main intermediate compounds, which can be further biodegraded into small molecules. The overall results obtained in this study are of valuable significance for the understanding of the behavior of PHE in soil and associated environment. Copyright © 2018 Elsevier Inc. All rights reserved.

  10. Ovariectomy results in inbred strain-specific increases in anxiety-like behavior in mice

    PubMed Central

    Schoenrock, Sarah Adams; Oreper, Daniel; Young, Nancy; Ervin, Robin Betsch; Bogue, Molly A.; Valdar, William; Tarantino, Lisa M.

    2017-01-01

    Women are at an increased risk for developing affective disorders during times of hormonal flux, including menopause when the ovaries cease production of estrogen. However, while all women undergo menopause, not all develop an affective disorder. Increased vulnerability can result from genetic predisposition, environmental factors and gene by environment interactions. In order to investigate interactions between genetic background and estrogen depletion, we performed bilateral ovariectomy, a surgical procedure that results in estrogen depletion and is thought to model the post-menopausal state, in a genetically defined panel of 37 inbred mouse strains. Seventeen days post-ovariectomy, we assessed behavior in two standard rodent assays of anxiety- and depressive-like behavior, the open field and forced swim tests. We detected a significant interaction between ovariectomy and genetic background on anxiety-like behavior in the open field. No strain specific effects of ovariectomy were observed in the forced swim assay. However, we did observe significant strain effects for all behaviors in both the open field and forced swim tests. This study is the largest to date to look at the effects of ovariectomy on behavior and provides evidence that ovariectomy interacts with genetic background to alter anxiety-like behavior in an animal model of menopause. PMID:27693591

  11. Model of an axially strained weakly guiding optical fiber modal pattern

    NASA Technical Reports Server (NTRS)

    Egalon, Claudio O.; Rogowski, Robert S.

    1992-01-01

    Axial strain can be determined by monitoring the modal pattern variation of an optical fiber. The results of a numerical model developed to calculate the modal pattern variation at the end of a weakly guiding optical fiber under axial strain is presented. Whenever an optical fiber is under stress, the optical path length, the index of refraction, and the propagation constants of each fiber mode change. In consequence, the modal phase term for the fields and the fiber output pattern are also modified. For multimode fibers, very complicated patterns result. The predicted patterns are presented, and an expression for the phase variation with strain is derived.

  12. Modelling and optimization of semi-solid processing of 7075 Al alloy

    NASA Astrophysics Data System (ADS)

    Binesh, B.; Aghaie-Khafri, M.

    2017-09-01

    The new modified strain-induced melt activation (SIMA) process presented by Binesh and Aghaie-Khafri was optimized using a response surface methodology to improve the thixotropic characteristics of semi-solid 7075 alloy. The responses, namely the average grain size and the shape factor, were considered as functions of three independent input variables: effective strain, isothermal holding temperature and time. Mathematical models for the responses were developed using the regression analysis technique, and the adequacy of the models was validated by the analysis of variance method. The calculated results correlated fairly well with the experiments. It was found that all the first- and second-order terms of the independent parameters and the interactive terms of the effective strain and holding time were statistically significant for the responses. In order to simultaneously optimize the responses, the desirable values for the effective strain, holding temperature and time were predicted to be 5.1, 609 °C and 14 min, respectively, when employing the desirability function approach. Based on the optimization results, a significant improvement in the average grain size and shape factor of the semi-solid slurry prepared by the new modified SIMA process was observed.

  13. Modeling constitutive behavior of a 15Cr-15Ni-2.2Mo-Ti modified austenitic stainless steel under hot compression using artificial neural network

    NASA Astrophysics Data System (ADS)

    Mandal, Sumantra

    2006-11-01

    ABSTRACT In this paper, an artificial neural network (ANN) model has been suggested to predict the constitutive flow behavior of a 15Cr-15Ni-2.2Mo-Ti modified austenitic stainless steel under hot deformation. Hot compression tests in the temperature range 850°C- 1250°C and strain rate range 10-3-102 s-1 were carried out. These tests provided the required data for training the neural network and for subsequent testing. The inputs of the neural network are strain, log strain rate and temperature while flow stress is obtained as output. A three layer feed-forward network with ten neurons in a single hidden layer and back-propagation learning algorithm has been employed. A very good correlation between experimental and predicted result has been obtained. The effect of temperature and strain rate on flow behavior has been simulated employing the ANN model. The results have been found to be consistent with the metallurgical trend. Finally, a monte carlo analiysis has been carried out to find out the noise sensitivity of the developed model.

  14. How biological background assumptions influence scientific risk evaluation of stacked genetically modified plants: an analysis of research hypotheses and argumentations.

    PubMed

    Rocca, Elena; Andersen, Fredrik

    2017-08-14

    Scientific risk evaluations are constructed by specific evidence, value judgements and biological background assumptions. The latter are the framework-setting suppositions we apply in order to understand some new phenomenon. That background assumptions co-determine choice of methodology, data interpretation, and choice of relevant evidence is an uncontroversial claim in modern basic science. Furthermore, it is commonly accepted that, unless explicated, disagreements in background assumptions can lead to misunderstanding as well as miscommunication. Here, we extend the discussion on background assumptions from basic science to the debate over genetically modified (GM) plants risk assessment. In this realm, while the different political, social and economic values are often mentioned, the identity and role of background assumptions at play are rarely examined. We use an example from the debate over risk assessment of stacked genetically modified plants (GM stacks), obtained by applying conventional breeding techniques to GM plants. There are two main regulatory practices of GM stacks: (i) regulate as conventional hybrids and (ii) regulate as new GM plants. We analyzed eight papers representative of these positions and found that, in all cases, additional premises are needed to reach the stated conclusions. We suggest that these premises play the role of biological background assumptions and argue that the most effective way toward a unified framework for risk analysis and regulation of GM stacks is by explicating and examining the biological background assumptions of each position. Once explicated, it is possible to either evaluate which background assumptions best reflect contemporary biological knowledge, or to apply Douglas' 'inductive risk' argument.

  15. Perceived exertion is as effective as the perceptual strain index in predicting physiological strain when wearing personal protective clothing.

    PubMed

    Borg, David N; Costello, Joseph T; Bach, Aaron J; Stewart, Ian B

    2017-02-01

    The perceptual strain index (PeSI) has been shown to overcome the limitations associated with the assessment of the physiological strain index (PSI), primarily the need to obtain a core body temperature measurement. The PeSI uses the subjective scales of thermal sensation and perceived exertion (RPE) to provide surrogate measures of core temperature and heart rate, respectively. Unfortunately, thermal sensation has shown large variability in providing an estimation of core body temperature. Therefore, the primary aim of this study was to determine if thermal comfort improved the ability of the PeSI to predict the PSI during exertional-heat stress. Eighteen healthy males (age: 23.5years; body mass: 79.4kg; maximal aerobic capacity: 57.2ml·kg -1 ·min -1 ) wore four different chemical/biological protective garments while walking on treadmill at a low (<325W) or moderate (326-499W) metabolic workload in environmental conditions equivalent to wet bulb globe temperatures 21, 30 or 37°C. Trials were terminated when heart rate exceeded 90% of maximum, when core body temperature reached 39°C, at 120min or due to volitional fatigue. Core body temperature, heart rate, thermal sensation, thermal comfort and RPE were recorded at 15min intervals and at termination. Multiple statistical methods were used to determine the most accurate perceptual predictor. Significant moderate relationships were observed between the PeSI (r=0.74; p<0.001), the modified PeSI (r=0.73; p<0.001) and unexpectedly RPE (r=0.71; p<0.001) with the PSI, respectively. The PeSI (mean bias: -0.8±1.5 based on a 0-10 scale; area under the curve: 0.887), modified PeSI (mean bias: -0.5±1.4 based on 0-10 scale; area under the curve: 0.886) and RPE (mean bias: -0.7±1.4 based on a 0-10 scale; area under the curve: 0.883) displayed similar predictive performance when participants experienced high-to-very high levels of physiological strain. Modifying the PeSI did not improve the subjective prediction of physiological strain. However, RPE provided an equally accurate prediction of physiological strain, particularly when high-to-very high levels of strain were observed. Therefore, given its predictive performance and user-friendliness, the evidence suggests that RPE in isolation is a practical and cost-effective tool able to estimate physiological strain during exertional-heat stress under these work conditions. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Evaluation of right heart function in a rat model using modified echocardiographic views.

    PubMed

    Bernardo, Ivan; Wong, James; Wlodek, Mary E; Vlahos, Ross; Soeding, Paul

    2017-01-01

    Echocardiography plays a major role in assessing cardiac function in animal models. We investigated use of a modified parasternal mid right-ventricular (MRV) and right ventricle (RV) outflow (RVOT) view, in assessing RV size and function, and the suitability of advanced 2D-strain analysis. 15 WKY rats were examined using transthoracic echocardiography. The left heart was assessed using standard short and long axis views. For the right ventricle a MRV and RVOT view were used to measure RV chamber and free wall area. 2D-strain analysis was applied to both ventricles using off-line analysis. RV chamber volume was determined by injection of 2% agarose gel, and RV free wall dissected and weighed. Echocardiography measurement was correlated with necropsy findings. The RV mid-ventricular dimension (R1) was 0.42±0.07cm and the right ventricular outflow tract dimension (R2) was 0.34±0.06cm, chamber end-diastolic area measurements were 0.38±0.09cm2 and 0.29±0.08cm2 for MRV and RVOT views respectively. RVOT and MRV chamber area correlated with gel mass. Doppler RV stroke volume was 0.32±0.08ml, cardiac output (CO) 110±27 ml.min-1 and RV free wall contractility assessed using 2D-strain analysis was demonstrated. We have shown that modified MRV and RVOT views can provide detailed assessment of the RV in rodents, with 2D-strain analysis of the RV free wall potentially feasible.

  17. Efficacy of Genetically Modified Bt Toxins Alone and in Combinations Against Pink Bollworm Resistant to Cry1Ac and Cry2Ab

    PubMed Central

    Tabashnik, Bruce E.; Fabrick, Jeffrey A.; Unnithan, Gopalan C.; Yelich, Alex J.; Masson, Luke; Zhang, Jie; Bravo, Alejandra; Soberón, Mario

    2013-01-01

    Evolution of resistance in pests threatens the long-term efficacy of insecticidal proteins from Bacillus thuringiensis (Bt) used in sprays and transgenic crops. Previous work showed that genetically modified Bt toxins Cry1AbMod and Cry1AcMod effectively countered resistance to native Bt toxins Cry1Ab and Cry1Ac in some pests, including pink bollworm (Pectinophora gossypiella). Here we report that Cry1AbMod and Cry1AcMod were also effective against a laboratory-selected strain of pink bollworm resistant to Cry2Ab as well as to Cry1Ab and Cry1Ac. Resistance ratios based on the concentration of toxin killing 50% of larvae for the resistant strain relative to a susceptible strain were 210 for Cry2Ab, 270 for Cry1Ab, and 310 for Cry1Ac, but only 1.6 for Cry1AbMod and 2.1 for Cry1AcMod. To evaluate the interactions among toxins, we tested combinations of Cry1AbMod, Cry1Ac, and Cry2Ab. For both the resistant and susceptible strains, the net results across all concentrations tested showed slight but significant synergism between Cry1AbMod and Cry2Ab, whereas the other combinations of toxins did not show consistent synergism or antagonism. The results suggest that the modified toxins might be useful for controlling populations of pink bollworm resistant to Cry1Ac, Cry2Ab, or both. PMID:24244692

  18. Modification of artificial sea water for the mass production of (+)-terrein by Aspergillus terreus strain PF26 derived from marine sponge Phakellia fusca.

    PubMed

    Yin, Y; Ding, Y; Feng, G; Li, J; Xiao, L; Karuppiah, V; Sun, W; Zhang, F; Li, Z

    2015-12-01

    (+)-Terrein shows multiple bioactivities, however, its mass production is a big challenge. Aspergillus terreus strain PF26 derived from South China Sea sponge Phakellia fusca has been cultured to produce (+)-terrein successfully, but artificial sea water (ASW) of high salinity used in the fermentation medium may cause the corrosion risk of metal bioreactor, which limits the fermentation on a large scale. In this study, we modified the components of ASW by removing NaCl and CaCl2 from the original formula, which reduced about 80% salinity of ASW. As a result, 7·56 g l(-1) (+)-terrein production was achieved in shake flask, which was 78·72% higher than using the original ASW, and the cultivation time was decreased from 24 to 15 days. Then, the modified ASW was used for the fermentation of A. terreus strain PF26 in a 500 l stirred bioreactor, consequently 2·5 g l(-1) of (+)-terrein production was achieved. The fermentation of marine micro-organisms always needs to use sea water or artificial sea water (ASW), which limits the fermentation on a large scale, as the high-salinity medium may cause the corrosion risk of bioreactor. In this study, the ASW formula is simplified to reduce the sea water salinity and improve the yield of (+)-terrein, finally, the modified ASW was successfully used for the mass production of (+)-terrein by A. terreus strain PF26 in a 500 l bioreactor. © 2015 The Society for Applied Microbiology.

  19. Electric field induced lattice strain in pseudocubic Bi(Mg{sub 1/2}Ti{sub 1/2})O{sub 3}-modified BaTiO{sub 3}-BiFeO{sub 3} piezoelectric ceramics

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

    Fujii, Ichiro, E-mail: ifujii@rins.ryukoku.ac.jp; Iizuka, Ryo; Ueno, Shintaro

    2016-04-25

    Contributions to the piezoelectric response in pseudocubic 0.3BaTiO{sub 3}-0.1Bi(Mg{sub 1/2}Ti{sub 1/2})O{sub 3}-0.6BiFeO{sub 3} ceramics were investigated by synchrotron X-ray diffraction under electric fields. All of the lattice strain determined from the 110, 111, and 200 pseudocubic diffraction peaks showed similar lattice strain hysteresis that was comparable to the bulk butterfly-like strain curve. It was suggested that the hysteresis of the lattice strain and the lack of anisotropy were related to the complex domain structure and the phase boundary composition.

  20. Antiplasmodial activity of new 4-aminoquinoline derivatives against chloroquine resistant strain.

    PubMed

    Sinha, Manish; Dola, Vasanth R; Agarwal, Pooja; Srivastava, Kumkum; Haq, Wahajul; Puri, Sunil K; Katti, Seturam B

    2014-07-15

    Emergence and spread of multidrug resistant strains of Plasmodium falciparum has severely limited the antimalarial chemotherapeutic options. In order to overcome the obstacle, a set of new side-chain modified 4-aminoquinolines were synthesized and screened against chloroquine-sensitive (3D7) and chloroquine-resistant (K1) strains of P. falciparum. The key feature of the designed molecules is the use of methylpiperazine linked α, β(3)- and γ-amino acids to generate novel side chain modified 4-aminoquinoline analogues. Among the evaluated compounds, 20c and 30 were found more potent than CQ against K1 and displayed a four-fold and a three-fold higher activity respectively, with a good selectivity index (SI=5846 and 11,350). All synthesized compounds had resistance index between 1.06 and >14.13 as against 47.2 for chloroquine. Biophysical studies suggested that this series of compounds act on heme polymerization target. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Bacterial resistance modifying tetrasaccharide agents from Ipomoea murucoides.

    PubMed

    Chérigo, Lilia; Pereda-Miranda, Rogelio; Gibbons, Simon

    2009-01-01

    As part of an ongoing project to identify oligosaccharides which modulate bacterial multidrug resistance, the CHCl(3)-soluble extract from flowers of a Mexican arborescent morning glory, Ipomoea murucoides, through preparative-scale recycling HPLC, yielded five lipophilic tetrasaccharide inhibitors of Staphylococcusaureus multidrug efflux pumps, murucoidins XII-XVI (1-5). The macrocyclic lactone-type structures for these linear hetero-tetraglycoside derivatives of jalapinolic acid were established by spectroscopic methods. These compounds were tested for in vitro antibacterial and resistance modifying activity against strains of Staphylococcus aureus possessing multidrug resistance efflux mechanisms. Only murucoidin XIV (3) displayed antimicrobial activity against SA-1199B (MIC 32microg/ml), a norfloxacin-resistant strain that over-expresses the NorA MDR efflux pump. The four microbiologically inactive (MIC>512microg/ml) tetrasaccharides increased norfloxacin susceptibility of this strain by 4-fold (8microg/ml from 32microg/ml) at concentrations of 25microg/ml, while murucoidin XIV (3) exerted the same potentiation effect at a concentration of 5microg/ml.

  2. The influence of climatically-driven surface loading variations on continental strain and seismicity

    NASA Astrophysics Data System (ADS)

    Craig, Tim; Calais, Eric; Fleitout, Luce; Bollinger, Laurent; Scotti, Oona

    2016-04-01

    In slowly deforming regions of plate interiors, secondary sources of stress and strain can result in transient deformation rates comparable to, or greater than, the background tectonic rates. Highly variable in space and time, these transients have the potential to influence the spatio-temporal distribution of seismicity, interfering with any background tectonic effects to either promote or inhibit the failure of pre-existing faults, and potentially leading to a clustered, or 'pulse-like', seismic history. Here, we investigate the ways in which the large-scale deformation field resulting from climatically-controlled changes in surface ice mass over the Pleistocene and Holocene may have influenced not only the seismicity of glaciated regions, but also the wider seismicity around the ice periphery. We first use a set of geodynamic models to demonstrate that a major pulse of seismic activity occurring in Fennoscandia, coincident with the time of end-glaciation, occurred in a setting where the contemporaneous horizontal strain-rate resulting from the changing ice mass, was extensional - opposite to the reverse sense of coseismic displacement accommodated on these faults. Therefore, faulting did not release extensional elastic strain that was building up at the time of failure, but compressional elastic strain that had accumulated in the lithosphere on timescales longer than the glacial cycle, illustrating the potential for a non-tectonic trigger to tap in to the background tectonic stress-state. We then move on to investigate the more distal influence that changing ice (and ocean) volumes may have had on the evolving strain field across intraplate Europe, how this is reflected in the seismicity across intraplate Europe, and what impact this might have on the paleoseismic record.

  3. Apolipoprotein A-IV constrains HPA and behavioral stress responsivity in a strain-dependent manner.

    PubMed

    Packard, Amy E B; Zhang, Jintao; Myers, Brent; Ko, Chih-Wei; Wang, Fei; Tso, Patrick; Ulrich-Lai, Yvonne M

    2017-12-01

    There is a critical gap in our knowledge of the mechanisms that govern interactions between daily life experiences (e.g., stress) and metabolic diseases, despite evidence that stress can have profound effects on cardiometabolic health. Apolipoprotein A-IV (apoA-IV) is a protein found in chylomicrons (lipoprotein particles that transport lipids throughout the body) where it participates in lipid handling and the regulation of peripheral metabolism. Moreover, apoA-IV is expressed in brain regions that regulate energy balance including the arcuate nucleus. Given that both peripheral and central metabolic processes are important modulators of hypothalamic-pituitary-adrenocortical (HPA) axis activity, the present work tests the hypothesis that apoA-IV activity affects stress responses. As emerging data suggests that apoA-IV actions can vary with background strain, we also explore the strain-dependence of apoA-IV stress regulation. These studies assess HPA axis, metabolic (hyperglycemia), and anxiety-related behavioral responses to psychogenic stress in control (wildtype) and apoA-IV-deficient (KO) mice on either the C57Bl/6J (C57) or 129×1/SvJ (129) background strain. The results indicate that apoA-IV KO increases post-stress corticosterone and anxiety-related behavior specifically in the 129 strain, and increases stress-induced hyperglycemia exclusively in the C57 strain. These data support the hypothesis that apoA-IV is a novel factor that limits stress reactivity in a manner that depends on genetic background. An improved understanding of the complex relationship among lipid homeostasis, stress sensitivity, and genetics is needed to optimize the development of personalized treatments for stress- and metabolism-related diseases. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Strain-specific induction of experimental autoimmune prostatitis (EAP) in mice.

    PubMed

    Jackson, Christopher M; Flies, Dallas B; Mosse, Claudio A; Parwani, Anil; Hipkiss, Edward L; Drake, Charles G

    2013-05-01

    Prostatitis, a clinical syndrome characterized by pelvic pain and inflammation, is common in adult males. Although several induced and spontaneous murine models of prostatitis have been explored, the role of genetic background on induction has not been well-defined. Using a standard methodology for the induction of experimental autoimmune prostatitis (EAP), we investigated both acute and chronic inflammation on several murine genetic backgrounds. In our colony, nonobese diabetic (NOD) mice evinced spontaneous prostatitis that was not augmented by immunization with rat prostate extract (RPE). In contrast, the standard laboratory strain Balb/c developed chronic inflammation in response to RPE immunization. Development of EAP in other strains was variable. These data suggest that Balb/c mice injected with RPE may provide a useful model for chronic prostatic inflammation. Copyright © 2012 Wiley Periodicals, Inc.

  5. Allelic Variation on Murine Chromosome 11 Modifies Host Inflammatory Responses and Resistance to Bacillus anthracis

    PubMed Central

    Terra, Jill K.; France, Bryan; Cote, Christopher K.; Jenkins, Amy; Bozue, Joel A.; Welkos, Susan L.; Bhargava, Ragini; Ho, Chi-Lee; Mehrabian, Margarete; Pan, Calvin; Lusis, Aldons J.; Davis, Richard C.; LeVine, Steven M.; Bradley, Kenneth A.

    2011-01-01

    Anthrax is a potentially fatal disease resulting from infection with Bacillus anthracis. The outcome of infection is influenced by pathogen-encoded virulence factors such as lethal toxin (LT), as well as by genetic variation within the host. To identify host genes controlling susceptibility to anthrax, a library of congenic mice consisting of strains with homozygous chromosomal segments from the LT-responsive CAST/Ei strain introgressed on a LT-resistant C57BL/6 (B6) background was screened for response to LT. Three congenic strains containing CAST/Ei regions of chromosome 11 were identified that displayed a rapid inflammatory response to LT similar to, but more severe than that driven by a LT-responsive allele of the inflammasome constituent NRLP1B. Importantly, increased response to LT in congenic mice correlated with greater resistance to infection by the Sterne strain of B. anthracis. The genomic region controlling the inflammatory response to LT was mapped to 66.36–74.67 Mb on chromosome 11, a region that encodes the LT-responsive CAST/Ei allele of Nlrp1b. However, known downstream effects of NLRP1B activation, including macrophage pyroptosis, cytokine release, and leukocyte infiltration could not fully explain the response to LT or the resistance to B. anthracis Sterne in congenic mice. Further, the exacerbated response in congenic mice is inherited in a recessive manner while the Nlrp1b-mediated response to LT is dominant. Finally, congenic mice displayed increased responsiveness in a model of sepsis compared with B6 mice. In total, these data suggest that allelic variation of one or more chromosome 11 genes in addition to Nlrp1b controls the severity of host response to multiple inflammatory stimuli and contributes to resistance to B. anthracis Sterne. Expression quantitative trait locus analysis revealed 25 genes within this region as high priority candidates for contributing to the host response to LT. PMID:22241984

  6. Comparative analysis of a modified ecolite method, the colicomplete method, and a most-probable-number method for detecting Escherichia coli in orange juice.

    PubMed

    Durbin, Gregory W; Salter, Robert

    2006-01-01

    The Ecolite High Volume Juice (HVJ) presence-absence method for a 10-ml juice sample was compared with the U.S. Food and Drug Administration Bacteriological Analytical Manual most-probable-number (MPN) method for analysis of artificially contaminated orange juices. Samples were added to Ecolite-HVJ medium and incubated at 35 degrees C for 24 to 48 h. Fluorescent blue results were positive for glucuronidase- and galactosidase-producing microorganisms, specifically indicative of about 94% of Escherichia coli strains. Four strains of E. coli were added to juices at concentrations of 0.21 to 6.8 CFU/ ml. Mixtures of enteric bacteria (Enterobacter plus Klebsiella, Citrobacter plus Proteus, or Hafnia plus Citrobacter plus Enterobacter) were added to simulate background flora. Three orange juice types were evaluated (n = 10) with and without the addition of the E. coli strains. Ecolite-HVJ produced 90 of 90 (10 of 10 samples of three juice types, each inoculated with three different E. coli strains) positive (blue-fluorescent) results with artificially contaminated E. coli that had MPN concentrations of <0.3 to 9.3 CFU/ml. Ten of 30 E. coli ATCC 11229 samples with MPN concentrations of <0.3 CFU/ml were identified as positive with Ecolite-HVJ. Isolated colonies recovered from positive Ecolite-HVJ samples were confirmed biochemically as E. coli. Thirty (10 samples each of three juice types) negative (not fluorescent) results were obtained for samples contaminated with only enteric bacteria and for uninoculated control samples. A juice manufacturer evaluated citrus juice production with both the Ecolite-HVJ and Colicomplete methods and recorded identical negative results for 95 20-ml samples and identical positive results for 5 20-ml samples artificially contaminated with E. coli. The Ecolite-HVJ method requires no preenrichment and subsequent transfer steps, which makes it a simple and easy method for use by juice producers.

  7. Large strain dynamic compression for soft materials using a direct impact experiment

    NASA Astrophysics Data System (ADS)

    Meenken, T.; Hiermaier, S.

    2006-08-01

    Measurement of strain rate dependent material data of low density low strength materials like polymeric foams and rubbers still poses challenges of a different kind to the experimental set up. For instance, in conventional Split Hopkinson Pressure Bar tests the impedance mismatch between the bars and the specimen makes strain measurement almost impossible. Application of viscoelastic bars poses new problems with wave dispersion. Also, maximum achievable strains and strain rates depend directly on the bar lengths, resulting in large experimental set ups in order to measure relevant data for automobile crash applications. In this paper a modified SHPB will be presented for testing low impedance materials. High strains can be achieved with nearly constant strain rate. A thin film stress measurement has been applied to the specimen/bar interfaces to investigate the initial sample ring up process. The process of stress homogeneity within the sample was investigated on EPDM and PU rubber.

  8. Changes in the Plasma Proteome Follows Chronic Opiate Administration In Simian Immunodeficiency Virus Infected Rhesus Macaques*

    PubMed Central

    Wiederin, Jayme L.; Yu, Fang; Donahoe, Robert M.; Fox, Howard S.; Ciborowski, Pawel; Gendelman, Howard E.

    2011-01-01

    Background Substantive plasma proteomic changes follow lentiviral infection and disease pathobiology. We posit that such protein alterations are modified during drug abuse, further serving to affect the disease. To this end, we investigated the effect of opiate administration on the plasma proteome of Indian-strain rhesus monkeys infected with simian immunodeficiency virus (SIV) strain smm9. Methods Whole blood was collected at 7 weeks prior to and 1.4 and 49 weeks after viral infection. Viral load, CD4+ T cell subsets, and plasma protein content were measured from monkeys that did or did not receive continuous opiate administrations. The plasma proteome was identified and quantified by isobaric tags for relative and absolute quantitation labeling (iTRAQ) and mass spectrometry. Results While substantive changes in plasma proteins were seen during SIV infection, the addition of opiates led to suppression of these changes as well as increased variance of the proteome. These changes demonstrate that opiates induce broad but variant immune suppression in SIV-infected monkeys. Conclusion The broad suppressive changes seen in plasma of SIV-infected monkeys likely reflect reduced multisystem immune homeostatic responses induced by opiates. Such occur as a consequence of complex cell-to-cell interactions operative between the virus and the host. We conclude that such changes in plasma proteomic profiling may be underappreciated and as such supports the need for improved clinical definitions. PMID:21821369

  9. Allelic Imbalance Is a Prevalent and Tissue-Specific Feature of the Mouse Transcriptome

    PubMed Central

    Pinter, Stefan F.; Colognori, David; Beliveau, Brian J.; Sadreyev, Ruslan I.; Payer, Bernhard; Yildirim, Eda; Wu, Chao-ting; Lee, Jeannie T.

    2015-01-01

    In mammals, several classes of monoallelic genes have been identified, including those subject to X-chromosome inactivation (XCI), genomic imprinting, and random monoallelic expression (RMAE). However, the extent to which these epigenetic phenomena are influenced by underlying genetic variation is unknown. Here we perform a systematic classification of allelic imbalance in mouse hybrids derived from reciprocal crosses of divergent strains. We observe that deviation from balanced biallelic expression is common, occurring in ∼20% of the mouse transcriptome in a given tissue. Allelic imbalance attributed to genotypic variation is by far the most prevalent class and typically is tissue-specific. However, some genotype-based imbalance is maintained across tissues and is associated with greater genetic variation, especially in 5′ and 3′ termini of transcripts. We further identify novel random monoallelic and imprinted genes and find that genotype can modify penetrance of parental origin even in the setting of large imprinted regions. Examination of nascent transcripts in single cells from inbred parental strains reveals that genes showing genotype-based imbalance in hybrids can also exhibit monoallelic expression in isogenic backgrounds. This surprising observation may suggest a competition between alleles and/or reflect the combined impact of cis- and trans-acting variation on expression of a given gene. Our findings provide novel insights into gene regulation and may be relevant to human genetic variation and disease. PMID:25858912

  10. FACTORS INFLUENCING AGE AND STRAIN-RELATED SUSCEPTIBILITY TO 3-METHYLCHOLANTHRENE CARCINOGENICITY

    EPA Science Inventory

    Fetal mice are more sensitive to chemical carcinogens than are adults. Further, some strains of mice are more susceptible to chemical carcinogens than others. We have been conducting studies to understand the interactions between age and genetic background underlying these suscep...

  11. A modified R-type bacteriocin specifically targeting Clostridium difficile prevents colonization of mice without affecting gut microbiota diversity.

    PubMed

    Gebhart, Dana; Lok, Stephen; Clare, Simon; Tomas, Myreen; Stares, Mark; Scholl, Dean; Donskey, Curtis J; Lawley, Trevor D; Govoni, Gregory R

    2015-03-24

    Clostridium difficile is a leading cause of nosocomial infections worldwide and has become an urgent public health threat requiring immediate attention. Epidemic lineages of the BI/NAP1/027 strain type have emerged and spread through health care systems across the globe over the past decade. Limiting person-to-person transmission and eradicating C. difficile, especially the BI/NAP1/027 strain type, from health care facilities are difficult due to the abundant shedding of spores that are impervious to most interventions. Effective prophylaxis for C. difficile infection (CDI) is lacking. We have genetically modified a contractile R-type bacteriocin ("diffocin") from C. difficile strain CD4 to kill BI/NAP1/027-type strains for this purpose. The natural receptor binding protein (RBP) responsible for diffocin targeting was replaced with a newly discovered RBP identified within a prophage of a BI/NAP1/027-type target strain by genome mining. The resulting modified diffocins (a.k.a. Avidocin-CDs), Av-CD291.1 and Av-CD291.2, were stable and killed all 16 tested BI/NAP1/027-type strains. Av-CD291.2 administered in drinking water survived passage through the mouse gastrointestinal (GI) tract, did not detectably alter the mouse gut microbiota or disrupt natural colonization resistance to C. difficile or the vancomycin-resistant Enterococcus faecium (VREF), and prevented antibiotic-induced colonization of mice inoculated with BI/NAP1/027-type spores. Given the high incidence and virulence of the pathogen, preventing colonization by BI/NAP1/027-type strains and limiting their transmission could significantly reduce the occurrence of the most severe CDIs. This modified diffocin represents a prototype of an Avidocin-CD platform capable of producing targetable, precision anti-C. difficile agents that can prevent and potentially treat CDIs without disrupting protective indigenous microbiota. Treatment and prevention strategies for bacterial diseases rely heavily on traditional antibiotics, which impose strong selection for resistance and disrupt protective microbiota. One consequence has been an upsurge of opportunistic pathogens, such as Clostridium difficile, that exploit antibiotic-induced disruptions in gut microbiota to proliferate and cause life-threatening diseases. We have developed alternative agents that utilize contractile bactericidal protein complexes (R-type bacteriocins) to kill specific C. difficile pathogens. Efficacy in a preclinical animal study indicates these molecules warrant further development as potential prophylactic agents to prevent C. difficile infections in humans. Since these agents do not detectably alter the indigenous gut microbiota or colonization resistance in mice, we believe they will be safe to administer as a prophylactic to block transmission in high-risk environments without rendering patients susceptible to enteric infection after cessation of treatment. Copyright © 2015 Gebhart et al.

  12. Comparative evaluation of two vaccine candidates against experimental leishmaniasis due to Leishmania major infection in four inbred mouse strains.

    PubMed

    Benhnini, Fouad; Chenik, Mehdi; Laouini, Dhafer; Louzir, Hechmi; Cazenave, Pierre André; Dellagi, Koussay

    2009-11-01

    Experimental leishmaniasis in BALB/c and C57BL/6 mice are the most investigated murine models that were used for the preclinical evaluation of Leishmania vaccine candidates. We have previously described two new inbred mouse strains named PWK and MAI issued from feral founders that also support the development of experimental leishmaniasis due to L. major. In this study, we sought to determine whether different mouse inbred strains generate concordant or discordant results when used to evaluate the potential of Leishmania proteins to protect against experimental leishmaniasis. To this end, two Leishmania proteins, namely, LACK (for Leishmania homolog of receptor for activated C kinase) and LmPDI (for L. major protein disulfide isomerase) were compared for their capacity to protect against experimental leishmaniasis in PWK, MAI, BALB/c, and C57BL/6 inbred mouse strains. Our data show that the capacity of Leishmania proteins to confer protection depends on the mouse strain used, stressing the important role played by the genetic background in shaping the immune response against the pathogen. These results may have important implications for the preclinical evaluation of candidate Leishmania vaccines: rather than using a single mouse strain, a panel of different inbred strains of various genetic backgrounds should be tested in parallel. The antigen that confers protection in the larger range of inbred strains may have better chances to be also protective in outbred human populations and should be selected for clinical trials.

  13. Comparison of two laryngeal tissue fiber constitutive models

    NASA Astrophysics Data System (ADS)

    Hunter, Eric J.; Palaparthi, Anil Kumar Reddy; Siegmund, Thomas; Chan, Roger W.

    2014-02-01

    Biological tissues are complex time-dependent materials, and the best choice of the appropriate time-dependent constitutive description is not evident. This report reviews two constitutive models (a modified Kelvin model and a two-network Ogden-Boyce model) in the characterization of the passive stress-strain properties of laryngeal tissue under tensile deformation. The two models are compared, as are the automated methods for parameterization of tissue stress-strain data (a brute force vs. a common optimization method). Sensitivity (error curves) of parameters from both models and the optimized parameter set are calculated and contrast by optimizing to the same tissue stress-strain data. Both models adequately characterized empirical stress-strain datasets and could be used to recreate a good likeness of the data. Nevertheless, parameters in both models were sensitive to measurement errors or uncertainties in stress-strain, which would greatly hinder the confidence in those parameters. The modified Kelvin model emerges as a potential better choice for phonation models which use a tissue model as one component, or for general comparisons of the mechanical properties of one type of tissue to another (e.g., axial stress nonlinearity). In contrast, the Ogden-Boyce model would be more appropriate to provide a basic understanding of the tissue's mechanical response with better insights into the tissue's physical characteristics in terms of standard engineering metrics such as shear modulus and viscosity.

  14. Constitutive Behavior and Deep Drawability of Three Aluminum Alloys Under Different Temperatures and Deformation Speeds

    NASA Astrophysics Data System (ADS)

    Panicker, Sudhy S.; Prasad, K. Sajun; Basak, Shamik; Panda, Sushanta Kumar

    2017-08-01

    In the present work, uniaxial tensile tests were carried out to evaluate the stress-strain response of AA2014, AA5052 and AA6082 aluminum alloys at four temperatures: 303, 423, 523 and 623 K, and three strain rates: 0.0022, 0.022 and 0.22 s-1. It was found that the Cowper-Symonds model was not a robust constitutive model, and it failed to predict the flow behavior, particularly the thermal softening at higher temperatures. Subsequently, a comparative study was made on the capability of Johnson-Cook (JC), modified Zerilli-Armstrong (m-ZA), modified Arrhenius (m-ARR) and artificial neural network (ANN) for modeling the constitutive behavior of all the three aluminum alloys under the mentioned strain rates and temperatures. Also, the improvement in formability of the materials was evaluated at an elevated temperature of 623 K in terms of cup height and maximum safe strains by conducting cylindrical cup deep drawing experiments under two different punch speeds of 4 and 400 mm/min. The cup heights increased during warm deep drawing due to thermal softening and increase in failure strains. Also, a small reduction in cup height was observed when the punch speed increased from 4 to 400 mm/min at 623 K. Hence, it was suggested to use high-speed deformation at elevated temperature to reduce both punch load and cycle time during the deep drawing process.

  15. Surface Topography and Mechanical Strain Promote Keratocyte Phenotype and Extracellular Matrix Formation in a Biomimetic 3D Corneal Model.

    PubMed

    Zhang, Wei; Chen, Jialin; Backman, Ludvig J; Malm, Adam D; Danielson, Patrik

    2017-03-01

    The optimal functionality of the native corneal stroma is mainly dependent on the well-ordered arrangement of extracellular matrix (ECM) and the pressurized structure. In order to develop an in vitro corneal model, it is crucial to mimic the in vivo microenvironment of the cornea. In this study, the influence of surface topography and mechanical strain on keratocyte phenotype and ECM formation within a biomimetic 3D corneal model is studied. By modifying the surface topography of materials, it is found that patterned silk fibroin film with 600 grooves mm -1 optimally supports cell alignment and ECM arrangement. Furthermore, treatment with 3% dome-shaped mechanical strain, which resembles the shape and mechanics of native cornea, significantly enhances the expression of keratocyte markers as compared to flat-shaped strain. Accordingly, a biomimetic 3D corneal model, in the form of a collagen-modified, silk fibroin-patterned construct subjected to 3% dome-shaped strain, is created. Compared to traditional 2D cultures, it supports a significantly higher expression of keratocyte and ECM markers, and in conclusion better maintains keratocyte phenotype, alignment, and fusiform cell shape. Therefore, the novel biomimetic 3D corneal model developed in this study serves as a useful in vitro 3D culture model to improve current 2D cultures for corneal studies. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines

    PubMed Central

    Mota, Rodrigo M; Moreira, João Luiz S; Souza, Marcelo R; Fátima Horta, M; Teixeira, Santuza MR; Neumann, Elisabeth; Nicoli, Jacques R; Nunes, Álvaro C

    2006-01-01

    Background The use of lactic acid bacteria as vehicles to delivery antigens to immunize animals is a promising issue. When genetically modified, these bacteria can induce a specific local and systemic immune response against selected pathogens. Gastric acid and bile salts tolerance, production of antagonistic substances against pathogenic microorganisms, and adhesive ability to gut epithelium are other important characteristics that make these bacteria useful for oral immunization. Results Bacteria isolated on de Man, Rogosa and Sharpe medium (MRS) from different gastrointestinal portions of broiler chicks were evaluated for their resistance to artificial gastric acid and bile salts, production of hydrogen peroxide, and cell surface hydrophobicity. Thirty-eight isolates were first typed at species level by PCR amplification of 16S-23S rRNA intergenic spacers using universal primers that anneal within 16S and 23S genes, followed by restriction digestion analyses of PCR amplicons (PCR-ARDRA). An expression cassette was assembled onto the pCR2.1-Topo vector by cloning the promoter, leader peptide, cell wall anchor and terminator sequences derived from the laminin binding S-layer protein gene of L. crispatus strain F5.7 (lbs gene). A sequence encoding the green fluorescent protein (GFP) was inserted as reporter gene, and an erythromycin resistance gene was added as selective marker. All constructs were able to express GFP in the cloning host E. coli XL1-Blue and different Lactobacillus strains as verified by FACS and laser scanning confocal microscopy. Conclusion Lactobacillus isolated from gastrointestinal tract of broiler chickens and selected for probiotic characteristics can be genetically modified by introducing an expression cassette into the lbs locus. The transformed bacteria expressed on its cell wall surface different fluorescent proteins used as reporters of promoter function. It is possible then that similar bacterial model expressing pathogen antigens can be used as live oral vaccines to immunize broilers against infectious diseases. PMID:16396687

  17. The Features of Fracture Behavior of an Aluminum-Magnesium Alloy AMg6 Under High-Rate Straining

    NASA Astrophysics Data System (ADS)

    Skripnyak, N. V.

    2015-09-01

    The results of investigation of fracture dynamics of rolled sheet specimens of an AMg6 alloy are presented for the range of strain rates from 10-3 to 103 s-1. It is found out that the presence of nanostructured surface layers on the thin AMg6 rolled sheets results in improved strength characteristics within the above range of strain rates. A modified model of a deforming medium is proposed to describe the plastic flow and fracture of the AMg6 alloy.

  18. Shear Melting of a Colloidal Glass

    NASA Astrophysics Data System (ADS)

    Eisenmann, Christoph; Kim, Chanjoong; Mattsson, Johan; Weitz, David A.

    2010-01-01

    We use confocal microscopy to explore shear melting of colloidal glasses, which occurs at strains of ˜0.08, coinciding with a strongly non-Gaussian step size distribution. For larger strains, the particle mean square displacement increases linearly with strain and the step size distribution becomes Gaussian. The effective diffusion coefficient varies approximately linearly with shear rate, consistent with a modified Stokes-Einstein relationship in which thermal energy is replaced by shear energy and the length scale is set by the size of cooperatively moving regions consisting of ˜3 particles.

  19. Stress-Strain Behavior of Cementitious Materials with Different Sizes

    PubMed Central

    Zhou, Jikai; Qian, Pingping; Chen, Xudong

    2014-01-01

    The size dependence of flexural properties of cement mortar and concrete beams is investigated. Bazant's size effect law and modified size effect law by Kim and Eo give a very good fit to the flexural strength of both cement mortar and concrete. As observed in the test results, a strong size effect in flexural strength is found in cement mortar than in concrete. A modification has been suggested to Li's equation for describing the stress-strain curve of cement mortar and concrete by incorporating two different correction factors, the factors contained in the modified equation being established empirically as a function of specimen size. A comparison of the predictions of this equation with test data generated in this study shows good agreement. PMID:24744688

  20. Technology for Elevated Temperature Tests of Structural Panels

    NASA Technical Reports Server (NTRS)

    Thornton, E. A.

    1999-01-01

    A technique for full-field measurement of surface temperature and in-plane strain using a single grid imaging technique was demonstrated on a sample subjected to thermally-induced strain. The technique is based on digital imaging of a sample marked by an alternating line array of La2O2S:Eu(+3) thermographic phosphor and chromium illuminated by a UV lamp. Digital images of this array in unstrained and strained states were processed using a modified spin filter. Normal strain distribution was determined by combining unstrained and strained grid images using a single grid digital moire technique. Temperature distribution was determined by ratioing images of phosphor intensity at two wavelengths. Combined strain and temperature measurements demonstrated on the thermally heated sample were DELTA-epsilon = +/- 250 microepsilon and DELTA-T = +/- 5 K respectively with a spatial resolution of 0.8 mm.

  1. Draft Genome Sequences of Two Janthinobacterium lividum Strains, Isolated from Pristine Groundwater Collected from the Oak Ridge Field Research Center

    PubMed Central

    Wu, Xiaoqin; Deutschbauer, Adam M.; Kazakov, Alexey E.; Wetmore, Kelly M.; Cwick, Bryson A.; Walker, Robert M.; Novichkov, Pavel S.; Arkin, Adam P.

    2017-01-01

    ABSTRACT We present here the draft genome sequences of two Janthinobacterium lividum strains, GW456P and GW458P, isolated from groundwater samples collected from a background site at the Oak Ridge Field Research Center. Production of a purple pigment by these two strains was observed when grown on diluted (1/10) LB agar plates. PMID:28663297

  2. Genome Sequence of Saccharomyces cerevisiae Strain Kagoshima No. 2, Used for Brewing the Japanese Distilled Spirit Shōchū.

    PubMed

    Mori, Kazuki; Kadooka, Chihiro; Masuda, Chika; Muto, Ai; Okutsu, Kayu; Yoshizaki, Yumiko; Takamine, Kazunori; Futagami, Taiki; Tamaki, Hisanori

    2017-10-12

    Here, we report a draft genome sequence of Saccharomyces cerevisiae strain Kagoshima no. 2, which is used for brewing shōchū, a traditional distilled spirit in Japan. The genome data will facilitate an understanding of the evolutional traits and genetic background related to the characteristic features of strain Kagoshima no. 2. Copyright © 2017 Mori et al.

  3. Genome Sequence of Saccharomyces cerevisiae Strain Kagoshima No. 2, Used for Brewing the Japanese Distilled Spirit Shōchū

    PubMed Central

    Mori, Kazuki; Kadooka, Chihiro; Masuda, Chika; Muto, Ai; Okutsu, Kayu; Yoshizaki, Yumiko; Takamine, Kazunori; Tamaki, Hisanori

    2017-01-01

    ABSTRACT Here, we report a draft genome sequence of Saccharomyces cerevisiae strain Kagoshima no. 2, which is used for brewing shōchū, a traditional distilled spirit in Japan. The genome data will facilitate an understanding of the evolutional traits and genetic background related to the characteristic features of strain Kagoshima no. 2. PMID:29025949

  4. Dihydroquercetin Does Not Affect Age-Dependent Increase in Blood Pressure and Angiotensin-Converting Enzyme Activity in the Aorta of Hypertensive Rats.

    PubMed

    Slashcheva, G A; Rykov, V A; Lobanov, A V; Murashev, A N; Kim, Yu A; Arutyunyan, T V; Korystova, A F; Kublik, L N; Levitman, M Kh; Shaposhnikona, V V; Korystov, Yu N

    2016-09-01

    We analyzed changes in angiotensin-converting enzyme activity in the aorta of hypertensive SHR rats against the background of age-related BP increase (from week 7 to 14) and the effect of dihydroquercetin on BP rise and angiotensin-converting enzyme activity. Normotensive WKY rats of the same age were used as the control. BP and activity of angiotensin-converting enzyme in the aorta of SHR rats increased with age. Dihydroquercetin in doses of 100 and 300 μg/kg per day had no effect on the increase of these parameters; dihydroquercetin administered to 14-week-old WKY rats in a dose of 300 μg/kg reduced activity of the angiotensin-converting enzyme. Thus, the early (7-14 weeks) increase in BP and angiotensin-converting enzyme activity in the aorta of SHR rats was not modified by flavonoids (dihydroquercetin) in contrast to other rat strains and humans, which is indicative of specificity of hypertension mechanism in SHR rats.

  5. Metastable Pluripotent States in NOD Mouse Derived ES Cells

    PubMed Central

    Hanna, Jacob; Markoulaki, Styliani; Mitalipova, Maisam; Cheng, Albert W.; Cassady, John P.; Staerk, Judith; Carey, Bryce W.; Lengner, Christopher J.; Foreman, Ruth; Love, Jennifer; Gao, Qing; Kim, Jongpil; Jaenisch, Rudolf

    2009-01-01

    Embryonic stem (ES) cells are isolated from the inner cell mass (ICM) of blastocysts, whereas epiblast stem cells (EpiSCs) are derived from the post-implantation epiblast and display a restricted developmental potential. Here we characterize pluripotent states in the non-obese diabetic (NOD) mouse strain, which prior to this study was considered “non-permissive” for ES cell derivation. We find that NOD stem cells can be stabilized by providing constitutive expression of Klf4 or c-Myc or small molecules that can replace these factors during in vitro reprogramming. The NOD ES and iPS cells appear “metastable”, as they acquire an alternative EpiSC-like identity after removal of the exogenous factors, while their reintroduction converts the cells back to ICM-like pluripotency. Our findings suggest that stem cells from different genetic backgrounds can assume distinct states of pluripotency in vitro, the stability of which is regulated by endogenous genetic determinants and can be modified by exogenous factors. PMID:19427283

  6. Susceptibility of human Plasmodium knowlesi infections to anti-malarials

    PubMed Central

    2013-01-01

    Background Evidence suggests that Plasmodium knowlesi malaria in Sarawak, Malaysian Borneo remains zoonotic, meaning anti-malarial drug resistance is unlikely to have developed in the absence of drug selection pressure. Therefore, adequate response to available anti-malarial treatments is assumed. Methods Here the ex vivo sensitivity of human P. knowlesi isolates in Malaysian Borneo were studied, using a WHO schizont maturation assay modified to accommodate the quotidian life cycle of this parasite. The in vitro sensitivities of P. knowlesi H strain adapted from a primate infection to in vitro culture (by measuring the production of Plasmodium lactate dehydrogenase) were also examined together with some assays using Plasmodium falciparum and Plasmodium vivax. Results Plasmodium knowlesi is uniformly highly sensitive to artemisinins, variably and moderately sensitive to chloroquine, and less sensitive to mefloquine. Conclusions Taken together with reports of clinical failures when P. knowlesi is treated with mefloquine, the data suggest that caution is required if using mefloquine in prevention or treatment of P. knowlesi infections, until further studies are undertaken. PMID:24245918

  7. The maternal-age-associated risk of congenital heart disease is modifiable.

    PubMed

    Schulkey, Claire E; Regmi, Suk D; Magnan, Rachel A; Danzo, Megan T; Luther, Herman; Hutchinson, Alayna K; Panzer, Adam A; Grady, Mary M; Wilson, David B; Jay, Patrick Y

    2015-04-09

    Maternal age is a risk factor for congenital heart disease even in the absence of any chromosomal abnormality in the newborn. Whether the basis of this risk resides with the mother or oocyte is unknown. The impact of maternal age on congenital heart disease can be modelled in mouse pups that harbour a mutation of the cardiac transcription factor gene Nkx2-5 (ref. 8). Here, reciprocal ovarian transplants between young and old mothers establish a maternal basis for the age-associated risk in mice. A high-fat diet does not accelerate the effect of maternal ageing, so hyperglycaemia and obesity do not simply explain the mechanism. The age-associated risk varies with the mother's strain background, making it a quantitative genetic trait. Most remarkably, voluntary exercise, whether begun by mothers at a young age or later in life, can mitigate the risk when they are older. Thus, even when the offspring carry a causal mutation, an intervention aimed at the mother can meaningfully reduce their risk of congenital heart disease.

  8. Antibiotic resistance pattern and evaluation of metallo-beta lactamase genes (VIM and IMP) in Pseudomonas aeruginosa strains producing MBL enzyme, isolated from patients with secondary immunodeficiency

    PubMed Central

    Shirani, Kiana; Ataei, Behrouz; Roshandel, Fardad

    2016-01-01

    Background: One of the most common causes of hospital-acquired secondary infections in hospitalized patients is Pseudomonas aeruginosa. The aim of this study is to evaluate the expression of IMP and VIM in Pseudomonas aeruginosa strains (carbapenem resistant and producer MBL enzyme) in patients with secondary immunodeficiency. Materials and Methods: In a cross sectional study, 96 patients with secondary immunodeficiency hospitalized in the Al-Zahra hospital were selected. Carbapenem resistant strains isolated and modified Hodge test was performed in order to confirm the presence of the metallo carbapenemase enzyme. Under the standard conditions they were sent to the central laboratory for investigating nosocomial infection Multiplex PCR. Results: Of 96 samples 28.1% were IMP positive, 5.2% VIM positive and 3.1% both VIM and IMP positive. The prevalence of multidrug resistance in the IMP and/or VIM negative samples was 29%, while all 5 VIM positive samples have had multidrug resistance. Also the prevalence of multi-drug resistance in IMP positive samples were 96.3% and in IMP and VIM positive samples were 100%. According to Fisher’s test, the prevalence of multi-drug resistance based on gene expression has significant difference (P < 0.001). Conclusion: Based on the results of this study it can be concluded that, a significant percentage of patients with secondary immunodeficiency that suffer nosocomial infections with multidrug resistance, especially Pseudomonas aeruginosa, are probably MBL-producing gene positive. Therefore the cause of infection should be considered in the hospital care system to identify their features, the presence of genes involved in the development of multi-drug resistance and antibiotic therapy. PMID:27563634

  9. Sae regulator factor impairs the response to photodynamic inactivation mediated by Toluidine blue in Staphylococcus aureus.

    PubMed

    Gándara, Lautaro; Mamone, Leandro; Dotto, Cristian; Buzzola, Fernanda; Casas, Adriana

    2016-12-01

    Photodynamic inactivation (PDI) involves the combined use of light and a photosensitizer, which, in the presence of oxygen, originates cytotoxic species capable of inactivating bacteria. Since the emergence of multi-resistant bacterial strains is becoming an increasing public health concern, PDI becomes an attractive choice. The aim of this work was to study the differential susceptibility to Toluidine blue (TB) mediated PDI (TB-PDI) of S. aureus mutants (RN6390 and Newman backgrounds) for different key regulators of virulence factors related to some extent to oxidative stress. Complete bacteria eradication of planktonic cultures of RN6390 S. aureus photosensitized with 13μM TB was obtained upon illumination with a low light dose of 4.2J/cm 2 from a non-coherent light source. Similarly, complete cell death was achieved applying 1.3μM TB and 19J/cm 2 light dose, showing that higher light doses can lead to equal cell death employing low photosensitizer concentrations. Interestingly, RN6390 in planktonic culture responded significantly better to TB-PDI than the Newman strain. We showed that deficiencies in rsbU, mgrA (transcription factors related to stress response) or agr (quorum sensing system involved in copper resistance to oxidative stress) did not modify the response of planktonic S. aureus to PDI. On the other hand, the two component system sae impaired the response to TB-PDI through a mechanism not related to the Eap adhesin. More severe conditions were needed to inactivate S. aureus biofilms (0.5mM TB, 157J/cm 2 laser light). In mutant sae biofilms, strain dependant differential susceptibilities are not noticed. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Outbreak of H3N2 Influenza at a US Military Base in Djibouti during the H1N1 Pandemic of 2009

    PubMed Central

    Cosby, Michael T.; Pimentel, Guillermo; Nevin, Remington L.; Fouad Ahmed, Salwa; Klena, John D.; Amir, Ehab; Younan, Mary; Browning, Robert; Sebeny, Peter J.

    2013-01-01

    Background Influenza pandemics have significant operational impact on deployed military personnel working in areas throughout the world. The US Department of Defense global influenza-like illness (ILI) surveillance network serves an important role in establishing baseline trends and can be leveraged to respond to outbreaks of respiratory illness. Objective We identified and characterized an operationally unique outbreak of H3N2 influenza at Camp Lemonnier, Djibouti occurring simultaneously with the H1N1 pandemic of 2009 [A(H1N1)pdm09]. Methods Enhanced surveillance for ILI was conducted at Camp Lemonnier in response to local reports of a possible outbreak during the A(H1N1)pdm09 pandemic. Samples were collected from consenting patients presenting with ILI (utilizing a modified case definition) and who completed a case report form. Samples were cultured and analyzed using standard real-time reverse transcriptase PCR (rt-RT-PCR) methodology and sequenced genetic material was phylogenetically compared to other published strains. Results rt-RT-PCR and DNA sequencing revealed that 25 (78%) of the 32 clinical samples collected were seasonal H3N2 and only 2 (6%) were A(H1N1)pdm09 influenza. The highest incidence of H3N2 occurred during the month of May and 80% of these were active duty military personnel. Phylogenetic analysis revealed that sequenced H3N2 strains were genetically similar to 2009 strains from the United States of America, Australia, and South east Asia. Conclusions This outbreak highlights challenges in the investigation of influenza among deployed military populations and corroborates the public health importance of maintaining surveillance systems for ILI that can be enhanced locally when needed. PMID:24339995

  11. Assessment of virulence diversity of methicillin-resistant Staphylococcus aureus strains with a Drosophila melanogaster infection model.

    PubMed

    Wu, Kaiyu; Conly, John; Surette, Michael; Sibley, Christopher; Elsayed, Sameer; Zhang, Kunyan

    2012-11-23

    Staphylococcus aureus strains with distinct genetic backgrounds have shown different virulence in animal models as well as associations with different clinical outcomes, such as causing infection in the hospital or the community. With S. aureus strains carrying diverse genetic backgrounds that have been demonstrated by gene typing and genomic sequences, it is difficult to compare these strains using mammalian models. Invertebrate host models provide a useful alternative approach for studying bacterial pathogenesis in mammals since they have conserved innate immune systems of biological defense. Here, we employed Drosophila melanogaster as a host model for studying the virulence of S. aureus strains. Community-associated methicillin-resistant S. aureus (CA-MRSA) strains USA300, USA400 and CMRSA2 were more virulent than a hospital-associated (HA)-MRSA strain (CMRSA6) and a colonization strain (M92) in the D. melanogaster model. These results correlate with bacterial virulence in the Caenorhabditis elegans host model as well as human clinical data. Moreover, MRSA killing activities in the D. melanogaster model are associated with bacterial replication within the flies. Different MRSA strains induced similar host responses in D. melanogaster, but demonstrated differential expression of common bacterial virulence factors, which may account for the different killing activities in the model. In addition, hemolysin α, an important virulence factor produced by S. aureus in human infections is postulated to play a role in the fly killing. Our results demonstrate that the D. melanogaster model is potentially useful for studying S. aureus pathogenicity. Different MRSA strains demonstrated diverse virulence in the D. melanogaster model, which may be the result of differing expression of bacterial virulence factors in vivo.

  12. Are job strain and sleep disturbances prognostic factors for neck/shoulder/arm pain? A cohort study of a general population of working age in Sweden.

    PubMed

    Rasmussen-Barr, E; Grooten, W J A; Hallqvist, J; Holm, L W; Skillgate, E

    2014-07-08

    To study whether job strain, that is, psychological job demands and decision latitude, and sleep disturbances among persons with occasional neck/shoulder/arm pain (NSAP) are prognostic factors for having experienced at least one episode of troublesome NSAP, and to determine whether sleep disturbances modify the association between job strain and troublesome NSAP. Prospective cohort study. Stockholm, Sweden. A population-based cohort of individuals with occasional NSAP (n=6979) who answered surveys in 2006 and 2010. Report of at least one episode of troublesome NSAP in 2010. The ORs for troublesome NSAP at follow-up were in individuals exposed to passive jobs 1.2 (95% CI 0.9 to 1.4); to active jobs 1.3 (95% CI 1.1 to 1.5); to high strain 1.5 (95% CI 1.0 to 2.4); to mild sleep disturbances 1.4 (95% CI 1.3 to 1.6) and to severe sleep disturbances 2.2 (95% CI 1.6 to 3.0). High strain and active jobs were associated with having experienced at least one episode of troublesome NSAP during the previous 6 months in persons with sleep disturbances, but not in individuals without sleep disturbances. Our results indicate that high strain, active jobs and sleep disturbances are prognostic factors that should be taken into account when implementing preventive measures to minimise the risk of troublesome NSAP among people of working age. We suggest that sleep disturbances may modify the association between high strain and troublesome NSAP. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  13. Are job strain and sleep disturbances prognostic factors for neck/shoulder/arm pain? A cohort study of a general population of working age in Sweden

    PubMed Central

    Rasmussen-Barr, E; Grooten, W J A; Hallqvist, J; Holm, L W; Skillgate, E

    2014-01-01

    Objective To study whether job strain, that is, psychological job demands and decision latitude, and sleep disturbances among persons with occasional neck/shoulder/arm pain (NSAP) are prognostic factors for having experienced at least one episode of troublesome NSAP, and to determine whether sleep disturbances modify the association between job strain and troublesome NSAP. Design Prospective cohort study. Setting Stockholm, Sweden. Participants A population-based cohort of individuals with occasional NSAP (n=6979) who answered surveys in 2006 and 2010. Outcome measures Report of at least one episode of troublesome NSAP in 2010. Results The ORs for troublesome NSAP at follow-up were in individuals exposed to passive jobs 1.2 (95% CI 0.9 to 1.4); to active jobs 1.3 (95% CI 1.1 to 1.5); to high strain 1.5 (95% CI 1.0 to 2.4); to mild sleep disturbances 1.4 (95% CI 1.3 to 1.6) and to severe sleep disturbances 2.2 (95% CI 1.6 to 3.0). High strain and active jobs were associated with having experienced at least one episode of troublesome NSAP during the previous 6 months in persons with sleep disturbances, but not in individuals without sleep disturbances. Conclusions Our results indicate that high strain, active jobs and sleep disturbances are prognostic factors that should be taken into account when implementing preventive measures to minimise the risk of troublesome NSAP among people of working age. We suggest that sleep disturbances may modify the association between high strain and troublesome NSAP. PMID:25005596

  14. Potential mechanisms of attenuation for rifampicin-passaged strains of Flavobacterium psychrophilum

    USDA-ARS?s Scientific Manuscript database

    Background: Flavobacterium psychrophilum is the etiologic agent of bacterial coldwater disease in salmonids. Earlier research showed that a rifampicin-passaged strain of F. psychrophilum (CSF 259-93B 17) caused no disease in rainbow trout (Oncorhynchus mykiss, Walbaum) while inducing a protective im...

  15. Virulence of two strains of Mycobacterium bovis in cattle following aerosol infection

    USDA-ARS?s Scientific Manuscript database

    Background Over the past two decades, highly virulent strains of Mycobacterium tuberculosis have emerged and spread rapidly in humans, suggesting a selective advantage based upon virulence. A similar scenario has not been described for Mycobacterium bovis infection in cattle (i.e., Bovine Tuberculos...

  16. PERSISTENT ANTIGENIC VARIATION OF INFLUENZA A VIRUSES AFTER INCOMPLETE NEUTRALIZATION IN OVO WITH HETEROLOGOUS IMMUNE SERUM

    PubMed Central

    Archetti, Italo; Horsfall, Frank L.

    1950-01-01

    Antigenic variants of influenza A virus strains emerge on serial passage in ovo in the presence of immune serum against different but related strains. An old laboratory strain (PR8) which had been through hundreds of animal passages was as readily modified by this procedure as recently recovered strains. Such variants apparently can be obtained at will and show antigenic patterns which are reproducible and appear to be predictable in terms of the immune serum used for their selection. Variant strains retain their new antigenic patterns on serial passage in ovo in the absence of immune serum. Limited serial passage in ovo of strains in the absence of immune serum did not result in the emergence of antigenic variants. Similarly, serial passages of strains in ovo in the presence of immune serum against widely different strains, which failed to show significant cross-neutralization, did not lead to the appearance of antigenic variants. PMID:14778924

  17. Fracture prediction using modified mohr coulomb theory for non-linear strain paths using AA3104-H19

    NASA Astrophysics Data System (ADS)

    Dick, Robert; Yoon, Jeong Whan

    2016-08-01

    Experiment results from uniaxial tensile tests, bi-axial bulge tests, and disk compression tests for a beverage can AA3104-H19 material are presented. The results from the experimental tests are used to determine material coefficients for both Yld2000 and Yld2004 models. Finite element simulations are developed to study the influence of materials model on the predicted earing profile. It is shown that only the YLD2004 model is capable of accurately predicting the earing profile as the YLD2000 model only predicts 4 ears. Excellent agreement with the experimental data for earing is achieved using the AA3104-H19 material data and the Yld2004 constitutive model. Mechanical tests are also conducted on the AA3104-H19 to generate fracture data under different stress triaxiality conditions. Tensile tests are performed on specimens with a central hole and notched specimens. Torsion of a double bridge specimen is conducted to generate points near pure shear conditions. The Nakajima test is utilized to produce points in bi-axial tension. The data from the experiments is used to develop the fracture locus in the principal strain space. Mapping from principal strain space to stress triaxiality space, principal stress space, and polar effective plastic strain space is accomplished using a generalized mapping technique. Finite element modeling is used to validate the Modified Mohr-Coulomb (MMC) fracture model in the polar space. Models of a hole expansion during cup drawing and a cup draw/reverse redraw/expand forming sequence demonstrate the robustness of the modified PEPS fracture theory for the condition with nonlinear forming paths and accurately predicts the onset of failure. The proposed methods can be widely used for predicting failure for the examples which undergo nonlinear strain path including rigid-packaging and automotive forming.

  18. A trans-acting Variant within the Transcription Factor RIM101 Interacts with Genetic Background to Determine its Regulatory Capacity.

    PubMed

    Read, Timothy; Richmond, Phillip A; Dowell, Robin D

    2016-01-01

    Most genetic variants associated with disease occur within regulatory regions of the genome, underscoring the importance of defining the mechanisms underlying differences in regulation of gene expression between individuals. We discovered a pair of co-regulated, divergently oriented transcripts, AQY2 and ncFRE6, that are expressed in one strain of Saccharomyces cerevisiae, ∑1278b, but not in another, S288c. By combining classical genetics techniques with high-throughput sequencing, we identified a trans-acting single nucleotide polymorphism within the transcription factor RIM101 that causes the background-dependent expression of both transcripts. Subsequent RNA-seq experiments revealed that RIM101 regulates many more targets in S288c than in ∑1278b and that deletion of RIM101 in both backgrounds abrogates the majority of differential expression between the strains. Strikingly, only three transcripts undergo a significant change in expression after swapping RIM101 alleles between backgrounds, implying that the differences in the RIM101 allele lead to a remarkably focused transcriptional response. However, hundreds of RIM101-dependent targets undergo a subtle but consistent shift in expression in the S288c RIM101-swapped strain, but not its ∑1278b counterpart. We conclude that ∑1278b may harbor a variant(s) that buffers against widespread transcriptional dysregulation upon introduction of a non-native RIM101 allele, emphasizing the importance of accounting for genetic background when assessing the impact of a regulatory variant.

  19. Phylogenetic distribution and expression of a penicillin-binding protein homologue, Ear and its significance in virulence of Staphylococcus aureus.

    PubMed

    Singh, Vineet K; Ring, Robert P; Aswani, Vijay; Stemper, Mary E; Kislow, Jennifer; Ye, Zhan; Shukla, Sanjay K

    2017-12-01

    Staphylococcus aureus is an opportunistic human pathogen that can cause serious infections in humans. A plethora of known and putative virulence factors are produced by staphylococci that collectively orchestrate pathogenesis. Ear protein (Escherichia coli ampicillin resistance) in S. aureus is an exoprotein in COL strain, predicted to be a superantigen, and speculated to play roles in antibiotic resistance and virulence. The goal of this study was to determine if expression of ear is modulated by single nucleotide polymorphisms in its promoter and coding sequences and whether this gene plays roles in antibiotic resistance and virulence. Promoter, coding sequences and expression of the ear gene in clinical and carriage S. aureus strains with distinct genetic backgrounds were analysed. The JE2 strain and its isogenic ear mutant were used in a systemic infection mouse model to determine the competiveness of the ear mutant.Results/Key findings. The ear gene showed a variable expression, with USA300FPR3757 showing a high-level expression compared to many of the other strains tested including some showing negligible expression. Higher expression was associated with agr type 1 but not correlated with phylogenetic relatedness of the ear gene based upon single nucleotide polymorphisms in the promoter or coding regions suggesting a complex regulation. An isogenic JE2 (USA300 background) ear mutant showed no significant difference in its growth, antibiotic susceptibility or virulence in a mouse model. Our data suggests that despite being highly expressed in a USA300 genetic background, Ear is not a significant contributor to virulence in that strain.

  20. Evolution of microstructure, strain and physical properties in oxide nanocomposite films

    DOE PAGES

    Chen, Aiping; Weigand, Marcus; Bi, Zhenxing; ...

    2014-06-24

    Using LSMO:ZnO nanocomposite films as a model system, we have researched the effect of film thickness on the physical properties of nanocomposites. It shows that strain, microstructure, as well as magnetoresistance strongly rely on film thickness. The magnetotransport properties have been fitted by a modified parallel connection channel model, which is in agreement with the microstructure evolution as a function of film thickness in nanocomposite films on sapphire substrates. The strain analysis indicates that the variation of physical properties in nanocomposite films on LAO is dominated by strain effect. These results confirm the critical role of film thickness on microstructures,more » strain states, and functionalities. Furthermore, it shows that one can use film thickness as a key parameter to design nanocomposites with optimum functionalities.« less

  1. Experimental Study and Modelling of Poly (Methyl Methacrylate) and Polycarbonate Compressive Behavior from Low to High Strain Rates

    NASA Astrophysics Data System (ADS)

    El-Qoubaa, Z.; Colard, L.; Matadi Boumbimba, R.; Rusinek, A.

    2018-06-01

    This paper concerns an experimental investigation of Polycarbonate and Poly (methyl methacrylate) compressive behavior from low to high strain rates. Experiments were conducted from 0.001/s to ≈ 5000/s for PC and from 0.001/s to ≈ 2000/s for PMMA. The true strain-stress behavior is established and analyzed at various stain rates. Both PC and PMMA mechanical behavior appears as known, to be strain rate and temperature dependent. The DSGZ model is selected for modelling the strain-stress curves while the yield stress is reproduced using the cooperative model and a modified Eyring equation based on Eyring first process theory. All the three models predictions are in agreement with experiments performed on PC and PMMA.

  2. Axial strain insensitivity of weakly guiding optical fibers

    NASA Technical Reports Server (NTRS)

    Egalon, Claudio O.; Rogowski, Robert S.

    1993-01-01

    A numerical model has been developed to calculate the modal phase shift of circular step index profile weakly guiding fibers under axial strain. Whenever an optical fiber is under stress, the optical path length, the index of refraction, and the propagation constants of each mode change. In consequence, the phase of each mode is also modified. A relationship for the modal phase shift is presented. This relation is applied to both single mode and two-mode fibers in order to determine the sensitivity characteristics of strained fibers. It was found that the phase shift is strongly dependent on the core refractive index, n(co). It was also found that it is possible to design fibers which are insensitive to axial strain. Practical applications of strain insensitive fibers are discussed.

  3. Experimental Study and Modelling of Poly (Methyl Methacrylate) and Polycarbonate Compressive Behavior from Low to High Strain Rates

    NASA Astrophysics Data System (ADS)

    El-Qoubaa, Z.; Colard, L.; Matadi Boumbimba, R.; Rusinek, A.

    2018-03-01

    This paper concerns an experimental investigation of Polycarbonate and Poly (methyl methacrylate) compressive behavior from low to high strain rates. Experiments were conducted from 0.001/s to ≈ 5000/s for PC and from 0.001/s to ≈ 2000/s for PMMA. The true strain-stress behavior is established and analyzed at various stain rates. Both PC and PMMA mechanical behavior appears as known, to be strain rate and temperature dependent. The DSGZ model is selected for modelling the strain-stress curves while the yield stress is reproduced using the cooperative model and a modified Eyring equation based on Eyring first process theory. All the three models predictions are in agreement with experiments performed on PC and PMMA.

  4. Genetic Contributors to Intergenerational CAG Repeat Instability in Huntington’s Disease Knock-In Mice

    PubMed Central

    Neto, João Luís; Lee, Jong-Min; Afridi, Ali; Gillis, Tammy; Guide, Jolene R.; Dempsey, Stephani; Lager, Brenda; Alonso, Isabel; Wheeler, Vanessa C.; Pinto, Ricardo Mouro

    2017-01-01

    Huntington’s disease (HD) is a neurodegenerative disorder caused by the expansion of a CAG trinucleotide repeat in exon 1 of the HTT gene. Longer repeat sizes are associated with increased disease penetrance and earlier ages of onset. Intergenerationally unstable transmissions are common in HD families, partly underlying the genetic anticipation seen in this disorder. HD CAG knock-in mouse models also exhibit a propensity for intergenerational repeat size changes. In this work, we examine intergenerational instability of the CAG repeat in over 20,000 transmissions in the largest HD knock-in mouse model breeding datasets reported to date. We confirmed previous observations that parental sex drives the relative ratio of expansions and contractions. The large datasets further allowed us to distinguish effects of paternal CAG repeat length on the magnitude and frequency of expansions and contractions, as well as the identification of large repeat size jumps in the knock-in models. Distinct degrees of intergenerational instability were observed between knock-in mice of six background strains, indicating the occurrence of trans-acting genetic modifiers. We also found that lines harboring a neomycin resistance cassette upstream of Htt showed reduced expansion frequency, indicative of a contributing role for sequences in cis, with the expanded repeat as modifiers of intergenerational instability. These results provide a basis for further understanding of the mechanisms underlying intergenerational repeat instability. PMID:27913616

  5. Field monitoring of plant-growth-promoting rhizobacteria by colony immunoblotting.

    PubMed

    Krishnen, Ganisan; Kecskés, Mihály L; Rose, Michael T; Geelan-Small, Peter; Amprayn, Khanok-on; Pereg, Lily; Kennedy, Ivan R

    2011-11-01

    Inoculant plant-growth-promoting bacteria are emerging as an important component of sustainable agriculture. There is a need to develop inexpensive methods for enumerating these organisms after their application in the field, to better understand their survival and impacts on yields. Immunoblotting is one potential method to measure viable cells, but the high cost of the conventionally used nylon membranes makes this method prohibitive. In this study, less expensive alternative materials such as filter papers, glossy photo papers, and transparencies for the purpose of colony immunoblotting were evaluated and the best substance was chosen for further studies. Whatman filter paper No. 541 combined with a 0.01 mol·L(-1) H(2)SO(4) rinsing step gave similar results to nylon membranes but <20% of the overall cost of the original colony immunoblotting assay. The application of the modified immunoblot method was tested on nonsterile clay soil samples that were spiked with high numbers (>10(7) CFU·g(-1)) of the plant-growth-promoting bacteria Pseudomonas fluorescens , Azospirillum brasilense , or Rhizobium leguminosarum . The modified protocol allowed the identification and recovery of over 50% of the inoculated cells of all three strains, amidst a background of the native soil microflora. Subsequently, the survival of P. fluorescens was successfully monitored for several months after application to field-grown rice at Jerilderie, New South Wales, Australia, thus validating the procedure.

  6. Genetic Contributors to Intergenerational CAG Repeat Instability in Huntington's Disease Knock-In Mice.

    PubMed

    Neto, João Luís; Lee, Jong-Min; Afridi, Ali; Gillis, Tammy; Guide, Jolene R; Dempsey, Stephani; Lager, Brenda; Alonso, Isabel; Wheeler, Vanessa C; Pinto, Ricardo Mouro

    2017-02-01

    Huntington's disease (HD) is a neurodegenerative disorder caused by the expansion of a CAG trinucleotide repeat in exon 1 of the HTT gene. Longer repeat sizes are associated with increased disease penetrance and earlier ages of onset. Intergenerationally unstable transmissions are common in HD families, partly underlying the genetic anticipation seen in this disorder. HD CAG knock-in mouse models also exhibit a propensity for intergenerational repeat size changes. In this work, we examine intergenerational instability of the CAG repeat in over 20,000 transmissions in the largest HD knock-in mouse model breeding datasets reported to date. We confirmed previous observations that parental sex drives the relative ratio of expansions and contractions. The large datasets further allowed us to distinguish effects of paternal CAG repeat length on the magnitude and frequency of expansions and contractions, as well as the identification of large repeat size jumps in the knock-in models. Distinct degrees of intergenerational instability were observed between knock-in mice of six background strains, indicating the occurrence of trans-acting genetic modifiers. We also found that lines harboring a neomycin resistance cassette upstream of Htt showed reduced expansion frequency, indicative of a contributing role for sequences in cis, with the expanded repeat as modifiers of intergenerational instability. These results provide a basis for further understanding of the mechanisms underlying intergenerational repeat instability. Copyright © 2017 by the Genetics Society of America.

  7. Investigation of creep by use of closed loop servo-hydraulic test system

    NASA Technical Reports Server (NTRS)

    Wu, H. C.; Yao, J. C.

    1981-01-01

    Creep tests were conducted by means of a closed loop servo-controlled materials test system. These tests are different from the conventional creep tests in that the strain history prior to creep may be carefully monitored. Tests were performed for aluminum alloy 6061-0 at 150 C and monitored by a PDP 11/04 minicomputer at a preset constant plastic-strain rate prehistory. The results show that the plastic-strain rate prior to creep plays a significant role in creep behavior. The endochronic theory of viscoplasticity was applied to describe the observed creep curves. The concepts of intrinsic time and strain rate sensitivity function are employed and modified according to the present observation.

  8. Model of an axially strained weakly guiding optical fiber modal pattern

    NASA Technical Reports Server (NTRS)

    Egalon, Claudio O.; Rogowski, Robert S.

    1991-01-01

    Axial strain may be determined by monitoring the modal pattern variation of an optical fiber. In this paper we present the results of a numerical model that has been developed to calculate the modal pattern variation at the end of a weakly guiding optical fiber under axial strain. Whenever an optical fiber is under stress, the optical path length, the index of refraction and the propagation constants of each fiber mode change. In consequence, the modal phase term of the fields and the fiber output pattern are also modified. For multimode fibers, very complicated patterns result. The predicted patterns are presented, and an expression for the phase variation with strain is derived.

  9. Adjustment of Trehalose Metabolism in Wine Saccharomyces cerevisiae Strains To Modify Ethanol Yields

    PubMed Central

    Rossouw, D.; Heyns, E. H.; Setati, M. E.; Bosch, S.

    2013-01-01

    The ability of Saccharomyces cerevisiae to efficiently produce high levels of ethanol through glycolysis has been the focus of much scientific and industrial activity. Despite the accumulated knowledge regarding glycolysis, the modification of flux through this pathway to modify ethanol yields has proved difficult. Here, we report on the systematic screening of 66 strains with deletion mutations of genes encoding enzymes involved in central carbohydrate metabolism for altered ethanol yields. Five of these strains showing the most prominent changes in carbon flux were selected for further investigation. The genes were representative of trehalose biosynthesis (TPS1, encoding trehalose-6-phosphate synthase), central glycolysis (TDH3, encoding glyceraldehyde-3-phosphate dehydrogenase), the oxidative pentose phosphate pathway (ZWF1, encoding glucose-6-phosphate dehydrogenase), and the tricarboxylic acid (TCA) cycle (ACO1 and ACO2, encoding aconitase isoforms 1 and 2). Two strains exhibited lower ethanol yields than the wild type (tps1Δ and tdh3Δ), while the remaining three showed higher ethanol yields. To validate these findings in an industrial yeast strain, the TPS1 gene was selected as a good candidate for genetic modification to alter flux to ethanol during alcoholic fermentation in wine. Using low-strength promoters active at different stages of fermentation, the expression of the TPS1 gene was slightly upregulated, resulting in a decrease in ethanol production and an increase in trehalose biosynthesis during fermentation. Thus, the mutant screening approach was successful in terms of identifying target genes for genetic modification in commercial yeast strains with the aim of producing lower-ethanol wines. PMID:23793638

  10. Accuracy of Different Implant Impression Techniques: Evaluation of New Tray Design Concept.

    PubMed

    Liu, David Yu; Cader, Fathima Nashmie; Abduo, Jaafar; Palamara, Joseph

    2017-12-29

    To evaluate implant impression accuracy with a new tray design concept in comparison to nonsplinted and splinted impression techniques for a 2-implant situation. A reference bar titanium framework was fabricated to fit on 2 parallel implants. The framework was used to generate a resin master model with 2 implants that fit precisely against the framework. Three impression techniques were evaluated: (1) nonsplinted, (2) splinted, and (3) nonsplinted with modified tray impressions. All the trays were fabricated from light-cured acrylic resin material with openings that corresponded to the implant impression copings. Ten impressions were taken for each technique using poly(vinyl siloxane) impression material. The impressions were poured with type IV dental stone to generate the test casts. A rosette strain gauge was bonded to the middle of the framework. As the framework retaining screws were tightened on each test cast, the developed strains were recorded until the completion of the tightening to 35 Ncm. The generated strains of the rosette strain gauge were used to calculate the maximum principal strain. A statistically significant difference was observed among the different impression techniques. The modified tray design impression technique was associated with the least framework strains, which indicates greater accuracy compared with the other techniques. There was no significant difference between the splinted and the nonsplinted impression techniques. The new tray design concept appeared to produce more accurate implant impressions than the other techniques. Despite the statistical difference among the impression techniques, the clinical significance of this difference is yet to be determined. © 2017 by the American College of Prosthodontists.

  11. Job strain and neck-shoulder symptoms: a prevalence study of women and men white-collar workers.

    PubMed

    Leroux, Isabelle; Brisson, Chantal; Montreuil, Sylvie

    2006-03-01

    Neck-shoulder symptoms are frequent among workers. Psychosocial factors at work have been associated with neck-shoulder symptoms, but few studies have examined job strain, the combined effect of high psychological demands (PD) and low decision latitude (DL). To examine the association between psychosocial factors at work and the prevalence of self-reported neck-shoulder symptoms among white-collar workers. In a cross-sectional study of 1543 white-collar workers, PD and DL at work were measured with Karasek's questionnaire. Prevalent cases were workers for whom neck-shoulder symptoms were present for >or=3 days during the previous 7 days and for whom pain intensity was greater than half the visual analogue scale. Gender and social support at work were evaluated as potential effect modifiers. Workers exposed to high job strain had a higher prevalence of neck-shoulder symptoms [adjusted prevalence ratio (PR): 1.54, 95% confidence interval (CI): 1.00-2.37]. No modifying effect of gender was observed in this association. The effect of job strain was stronger in workers with low social support (adjusted PR: 1.84, 95% CI: 0.92-3.68). These associations tended to be stronger and/or more precise when using alternative exposures and case definition. Namely, a stronger job strain effect was observed when a tertile cut-off was used to classify exposure (adjusted PR: 2.47, 95% CI: 1.15-5.32). These results suggest that primary prevention of neck-shoulder symptoms among white-collar workers should consider the exposure to job strain, especially when workers are exposed to low social support at work.

  12. Elaboration of highly hydrophobic polymeric surface--a potential strategy to reduce the adhesion of pathogenic bacteria?

    PubMed

    Poncin-Epaillard, F; Herry, J M; Marmey, P; Legeay, G; Debarnot, D; Bellon-Fontaine, M N

    2013-04-01

    Different polymeric surfaces have been modified in order to reach a high hydrophobic character, indeed the superhydrophobicity property. For this purpose, polypropylene and polystyrene have been treated by RF or μwaves CF4 plasma with different volumes, the results were compared according to the density of injected power. The effect of pretreatment such as mechanical abrasion or plasma activation was also studied. The modified surfaces were shown as hydrophobic, or even superhydrophobic depending of defects density. They were characterized by measurement of wettability and roughness at different scales, i.e. macroscopic, mesoscopic and atomic. It has been shown that a homogeneous surface at the macroscopic scale could be heterogeneous at lower mesoscopic scale. This was associated with the crystallinity of the material. The bioadhesion tests were performed with Gram positive and negative pathogenic strains: Listeria monocytogenes, Pseudomonas aeruginosa and Hafnia alvei. They have demonstrated an antibacterial efficiency of very hydrophobic and amorphous PS treated for all strains tested and a strain-dependent efficiency with modified PP surface being very heterogeneous at the mesoscopic scale. Thus, these biological results pointed out not only the respective role of the surface chemistry and topography in bacterial adhesion, but also the dependence on the peaks and valley distribution at bacteria dimension scale. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Nationwide survey of the development of drug resistance in the pediatric field in 2000-2001, 2004, 2007, 2010, and 2012: evaluation of the changes in drug sensitivity of Haemophilus influenzae and patients' background factors.

    PubMed

    Shiro, Hiroyuki; Sato, Yoshitake; Toyonaga, Yoshikiyo; Hanaki, Hideaki; Sunakawa, Keisuke

    2015-04-01

    The Drug-Resistant Pathogen Surveillance Group in Pediatric Infectious Disease has conducted surveillance of pediatric patients with respiratory tract infections, meningitis, and sepsis five times (in 2000-2001 [period 1], 2004 [period 2], 2007 [period 3], 2010 [period 4], and 2012 [period 5]). With respect to the clinically isolated Haemophilus influenzae, the drug susceptibility, the frequency of drug-resistant strains, and patients' background factors in each period have already been reported. Here we evaluate trends in the development of drug resistance in H. influenzae, and the relationship between the development of drug resistance and patients' background factors in the aforementioned five periods. H. influenzae derived from pediatric patients with respiratory tract infections that had been previously collected (period 1, 448 isolates; period 2, 376 isolates; period 3, 386 isolates; period 4, 484 isolates; and period 5, 411 isolates) were analyzed. The proportions of ß-lactamase-nonproducing ampicillin (ABPC)-intermediate resistant (BLNAI) strains + β-lactamase-nonproducing ABPC-resistant (BLNAR) strains were 28.8% in period 1, 59.3% in period 2, 61.1% in period 3, 58.1% in period 4, and 63.5% in period 5, showing a rapid increase from period 1 to period 2 followed by an almost constant rate of approximately 60%. The proportion of ß-lactamase-producing ABPC-resistant (BLPAR) strains + ß-lactamase-producing clavulanic acid/amoxicillin-resistant (BLPACR) strains was 4.4% in period 3, which was somewhat low; however, there were no significant changes in the proportions of these strains, which ranged between 6.4% and 8.7% throughout the surveillance period except for period 3. The drugs whose MIC90 values against BLNAR strains were low throughout the surveillance included piperacillin (0.25 μg/mL) and tazobactam/piperacillin (0.125-0.25 μg/mL) in the penicillins; cefditoren and ceftriaxone (0.25-0.5 μg/mL for both) in the cephems; meropenem (0.5-1 μg/mL) and tebipenem (1 μg/mL) in the carbapenems; and levofloxacin, tosufloxacin, and garenoxacin (≤ 0.06 μg/mL for all) and norfloxacin (0.06-0.125 μg/mL) in the quinolones. We investigated the relationship between the frequency of BLNAS strains/BLNAI + BLNAR strains and patients' background factors in each surveillance period. Significant differences were shown on age category (< 3 years or ≥ 3 years) in all periods except period 4, and the presence/absence of prior administration of antimicrobial agents within one month in period 2 and period 3. In all periods, the frequency of BLNAI + BLNAR strains were higher in patients aged < 3 years than in patients aged ≥ 3 years, and were also higher in patients with presence of prior treatment than in patients without prior treatment. We consider that it is important to promote the proper use of antimicrobial agents by conducting surveillance continuously in the future to clarify the relationship between the development of drug resistance in H. influenzae and patients' background factors and provide those information to clinical setting. Copyright © 2015. Published by Elsevier Ltd.

  14. Draft Genome Sequences of Two Janthinobacteriumlividum Strains, Isolated from Pristine Groundwater Collected from the Oak Ridge Field Research Center.

    PubMed

    Wu, Xiaoqin; Deutschbauer, Adam M; Kazakov, Alexey E; Wetmore, Kelly M; Cwick, Bryson A; Walker, Robert M; Novichkov, Pavel S; Arkin, Adam P; Chakraborty, Romy

    2017-06-29

    We present here the draft genome sequences of two Janthinobacterium lividum strains, GW456P and GW458P, isolated from groundwater samples collected from a background site at the Oak Ridge Field Research Center. Production of a purple pigment by these two strains was observed when grown on diluted (1/10) LB agar plates. Copyright © 2017 Wu et al.

  15. Genetic Characterization of Zika Virus Strains: Geographic Expansion of the Asian Lineage

    DTIC Science & Technology

    2012-02-28

    Genetic Characterization of Zika Virus Strains: Geographic Expansion of the Asian Lineage Andrew D. Haddow1*, Amy J. Schuh1, Chadwick Y. Yasuda2...Medical Research Unit, No. 2, Phnom Penh, Cambodia, 3 National Dengue Control Program, Phnom Penh, Cambodia Abstract Background: Zika virus (ZIKV) is a...underreported. Citation: Haddow AD, Schuh AJ, Yasuda CY, Kasper MR, Heang V, et al. (2012) Genetic Characterization of Zika Virus Strains: Geographic

  16. Occurrence of severe gastroenteritis in pups after canine parvovirus vaccine administration: a clinical and laboratory diagnostic dilemma.

    PubMed

    Decaro, Nicola; Desario, Costantina; Elia, Gabriella; Campolo, Marco; Lorusso, Alessio; Mari, Viviana; Martella, Vito; Buonavoglia, Canio

    2007-01-26

    A total of 29 faecal samples collected from dogs with diarrhoea following canine parvovirus (CPV) vaccination were tested by minor groove binder (MGB) probe assays for discrimination between CPV vaccine and field strains and by diagnostic tests for detection of other canine pathogens. Fifteen samples tested positive only for CPV field strains; however, both vaccine and field strains were detected in three samples. Eleven samples were found to contain only the vaccine strain, although eight of them tested positive for other pathogens of dogs. Only three samples were found to contain the vaccine strain without evidence of canine pathogens. The present study confirms that most cases of parvovirus-like disease occurring shortly after vaccination are related to infection with field strains of canine parvovirus type 2 (CPV-2) rather than to reversion to virulence of the modified live virus contained in the vaccine.

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

    Lee, Jaekwang; Huang, Jingsong; Sumpter, Bobby G.

    Compared with their bulk counterparts, 2D materials can sustain much higher elastic strain at which optical quantities such as bandgaps and absorption spectra governing optoelectronic device performance can be modified with relative ease. Using first-principles density functional theory and quasiparticle GW calculations, we demonstrate how uniaxial tensile strain can be utilized to optimize the electronic and optical properties of transition metal dichalcogenide lateral (in-plane) heterostructures such as MoX 2/WX 2 (X = S, Se, Te). We find that these lateral-type heterostructures may facilitate efficient electron–hole separation for light detection/harvesting and preserve their type II characteristic up to 12% of uniaxialmore » strain. Based on the strain-dependent bandgap and band offset, we show that uniaxial tensile strain can significantly increase the power conversion efficiency of these lateral heterostructures. Our results suggest that these strain-engineered lateral heterostructures are promising for optimizing optoelectronic device performance by selectively tuning the energetics of the bandgap.« less

  18. Nationwide survey of the development of drug resistance in the pediatric field in 2007, 2010, and 2012: drug sensitivity of Haemophilus influenzae serotype b strain in Japan.

    PubMed

    Baba, Hiroaki; Sato, Yoshitake; Toyonaga, Yoshikiyo; Hanaki, Hideaki; Sunakawa, Keisuke

    2015-04-01

    Based on the results of surveillance in the pediatric field conducted in 2007, 2010, and 2012, we examined the frequency of Haemophilus influenzae serotype b (Hib) strains, the susceptibility for Hib strains to various types of antimicrobial agent, and the relations to patients' background factors. Among all of Haemophilus influenzae, the frequency of Hib strains was 3.6% (14/386 strains) in 2007, 4.8% (23/484 strains) in 2010, 1.2% (5/411 strains) in 2012, and decreasing in 2012. Hib strains were isolated in patients with the following infections: nine patients with respiratory tract infections (upper respiratory tract infection, bronchitis, and pneumonia), three patients with sepsis, one patient with meningitis, and one patient with purulent inflammation of a tendon sheath in 2007; 11 patients with respiratory tract infections (upper respiratory tract infection, bronchitis, and pneumonia), four patients with sepsis, and eight patients with meningitis in 2010, demonstrating a relatively high frequency in patients with invasive infections. However, in 2012, Hib strains were isolated in only four patients with respiratory tract infections (upper respiratory tract infection) and one patient with bronchial asthma. Evaluation of background factors with pediatric patients in whom Hib strains were isolated showed that approximately 70% were male; majority was children under three years of age; and higher detection rates were also related to the background of patients who were attendant to daycare center, had siblings, had received no antimicrobial agents within the previous one month before collecting specimens. Throughout the surveillance between 2007 and 2012, antimicrobial agents with all phases' MICs ≤ 1 μg/mL were cefditoren, cefcapene, and cefteram in the oral β-lactams; tazobactam/piperacillin, ceftriaxone, cefotaxime, and meropenem in the injectable β-lactams; azithromycin in the macrolide; and levofloxacin in the quinolone. After 2010, MIC ranges were evaluated for tebipenem in the oral β-lactam (≤ 0.063-0.25 μg/mL), doripenem in the injectable β-lactam (≤ 0.063-0.5 μg/mL), and tosufloxacin in the quinolone (≤ 0.063 μg/mL). Throughout the surveillance, Hib strains were highly susceptible to the above mentioned antimicrobial agents, and there was no significant change in the susceptibility. Copyright © 2014 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  19. Whole Genome Analysis of a Wine Yeast Strain

    PubMed Central

    Hauser, Nicole C.; Fellenberg, Kurt; Gil, Rosario; Bastuck, Sonja; Hoheisel, Jörg D.

    2001-01-01

    Saccharomyces cerevisiae strains frequently exhibit rather specific phenotypic features needed for adaptation to a special environment. Wine yeast strains are able to ferment musts, for example, while other industrial or laboratory strains fail to do so. The genetic differences that characterize wine yeast strains are poorly understood, however. As a first search of genetic differences between wine and laboratory strains, we performed DNA-array analyses on the typical wine yeast strain T73 and the standard laboratory background in S288c. Our analysis shows that even under normal conditions, logarithmic growth in YPD medium, the two strains have expression patterns that differ significantly in more than 40 genes. Subsequent studies indicated that these differences correlate with small changes in promoter regions or variations in gene copy number. Blotting copy numbers vs. transcript levels produced patterns, which were specific for the individual strains and could be used for a characterization of unknown samples. PMID:18628902

  20. A modified Elek test for detection of toxigenic corynebacteria in the diagnostic laboratory.

    PubMed

    Engler, K H; Glushkevich, T; Mazurova, I K; George, R C; Efstratiou, A

    1997-02-01

    The detection of toxigenicity among Corynebacterium diphtheriae and Corynebacterium ulcerans strains is the most important test for the microbiological diagnosis of diphtheria. Difficulties with current methods, in particular the Elek test, are well documented. We therefore describe a modified Elek test which provides an accurate result after only 16 h of incubation, in contrast to 48 h for the conventional test.

  1. Bacterial migration along solid surfaces.

    PubMed Central

    Harkes, G; Dankert, J; Feijen, J

    1992-01-01

    An in vitro system was developed to study the migration of uropathogenic Escherichia coli strains. In this system an aqueous agar gel is placed against a solid surface, allowing the bacteria to migrate along the gel/solid surface interface. Bacterial strains as well as solid surfaces were characterized by means of water contact angle and zeta potential measurements. When glass was used as the solid surface, significantly different migration times for the strains investigated were observed. Relationships among the observed migration times of six strains, their contact angles, and their zeta potentials were found. Relatively hydrophobic strains exhibited migration times shorter than those of hydrophilic strains. For highly negatively charged strains shorter migration times were found than were found for less negatively charged strains. When the fastest-migrating strain with respect to glass was allowed to migrate along solid surfaces differing in hydrophobicity and charge, no differences in migration times were found. Our findings indicate that strategies to prevent catheter-associated bacteriuria should be based on inhibition of bacterial growth rather than on modifying the physicochemical character of the catheter surface. PMID:1622217

  2. Dynamic toughness in elastic nonlinear viscous solids

    NASA Astrophysics Data System (ADS)

    Tang, S.; Guo, T. F.; Cheng, L.

    2009-02-01

    This work addresses the interrelationship among dissipative mechanisms—material separation in the fracture process zone (FPZ), nonelastic deformation in the surrounding background material and kinetic energy—and how they affect the macroscopic dynamic fracture toughness as well as the limiting crack speed in strain rate sensitive materials. To this end, a micromechanics-based model for void growth in a nonlinear viscous solid is incorporated into a microporous strip of cell elements that forms the FPZ. The latter is surrounded by background material described by conventional constitutive relations. In the first part of the paper, the background material is assumed to be purely elastic. Here, the computed dynamic fracture toughness is a convex function of crack velocity. In the second part, the background material as well as the FPZ are described by similar rate-sensitivity parameters. Voids grow in the strain rate strengthened FPZ as the crack velocity increases. Consequently, the work of separation increases. By contrast, the inelastic dissipation in the background material appears to be a concave function of crack velocity. At the lower crack velocity regime, where dissipation in the background material is dominant, toughness decreases as crack velocity increases. At high crack velocities, inelastic deformation enhanced by the inertial force can cause a sharp increase in toughness. Here, the computed toughness increases rapidly with crack velocity. There exist regimes where the toughness is a non-monotonic function of the crack velocity. Two length scales—the width of the FPZ and the ratio of the shear wave speed to the reference strain rate—can be shown to strongly affect the dynamic fracture toughness. Our computational simulations can predict experimental data for fracture toughness vs. crack velocity reported in several studies for amorphous polymeric materials.

  3. Association of Recurrent Furunculosis with Panton-Valentine Leukocidin and the Genetic Background of Staphylococcus aureus▿ †

    PubMed Central

    Masiuk, Helena; Kopron, Katarzyna; Grumann, Dorothee; Goerke, Christiane; Kolata, Julia; Jursa-Kulesza, Joanna; Giedrys-Kalemba, Stefania; Bröker, Barbara M.; Holtfreter, Silva

    2010-01-01

    Staphylococcus aureus is a major cause of skin and soft tissue infections, such as furuncles, carbuncles, and abscesses, but it also frequently colonizes the human skin and mucosa without causing clinical symptoms. Panton-Valentine leukocidin (PVL) is a pore-forming toxin that has been associated with soft tissue infections and necrotizing pneumonia. We have compared the genotypes, virulence gene repertoires, and phage patterns of 74 furunculosis isolates with those of 108 control strains from healthy nasal carriers. The large majority of furunculosis strains were methicillin sensitive. Clonal cluster (CC) 121 (CC121) and CC22 accounted for 70% of the furunculosis strains but for only 8% of the nasal isolates. The PVL-encoding genes luk-PV were detected in 85% of furunculosis strains, while their prevalence among colonizing S. aureus strains was below 1%. luk-PV genes were distributed over several lineages (CCs 5, 8, 22, 30, and 121 and sequence type 59). Even within the same lineages, luk-PV-positive phages characterized furunculosis strains, while their luk-PV-negative variants were frequent among nasal strains. The very tight epidemiological linkage between luk-PV and furunculosis, which could be separated from the genetic background of the S. aureus strain as well as from the gene makeup of the luk-PV-transducing phage, lends support to the notion of an important role for PVL in human furunculosis. These results make a case for the determination of luk-PV in recurrent soft tissue infections with methicillin-sensitive as well as methicillin-resistant S. aureus. PMID:20200289

  4. Investigating the Mechanisms and Potential of Silk Fiber Metallization

    DTIC Science & Technology

    2013-09-30

    in an ALD process, when using the modified metal infiltration process as outlined by Lee et al., the titanium isopropoxide (TIP) precursor...these fibers exhibited >2-fold increase in strain to breakage, and >4.5-fold increase in strength when infiltrated with zinc, titanium , or aluminum...fibers exhibited >2-fold increase in strain to breakage, and >4.5-fold increase in strength when infiltrated with zinc, titanium , or aluminum

  5. L-arabinose fermenting yeast

    DOEpatents

    Zhang, Min; Singh, Arjun; Knoshaug, Eric; Franden, Mary Ann; Jarvis, Eric; Suominen, Pirkko

    2010-12-07

    An L-arabinose utilizing yeast strain is provided for the production of ethanol by introducing and expressing bacterial araA, araB and araD genes. L-arabinose transporters are also introduced into the yeast to enhance the uptake of arabinose. The yeast carries additional genomic mutations enabling it to consume L-arabinose, even as the only carbon source, and to produce ethanol. Methods of producing ethanol include utilizing these modified yeast strains. ##STR00001##

  6. Microorganisms having enhanced resistance to acetate and methods of use

    DOEpatents

    Brown, Steven D; Yang, Shihui

    2014-10-21

    The present invention provides isolated or genetically modified strains of microorganisms that display enhanced resistance to acetate as a result of increased expression of a sodium proton antiporter. The present invention also provides methods for producing such microbial strains, as well as related promoter sequences and expression vectors. Further, the present invention provides methods of producing alcohol from biomass materials by using microorganisms with enhanced resistance to acetate.

  7. Learning Strategy Selection in the Water Maze and Hippocampal CREB Phosphorylation Differ in Two Inbred Strains of Mice

    ERIC Educational Resources Information Center

    Sung, Jin-Young; Goo, June-Seo; Lee, Dong-Eun; Jin, Da-Qing; Bizon, Jennifer L.; Gallagher, Michela; Han, Jung-Soo

    2008-01-01

    Learning strategy selection was assessed in two different inbred strains of mice, C57BL/6 and DBA/2, which are used for developing genetically modified mouse models. Male mice received a training protocol in a water maze using alternating blocks of visible and hidden platform trials, during which mice escaped to a single location. After training,…

  8. Phenotypic characterisation of Saccharomyces spp. for tolerance to 1-butanol.

    PubMed

    Zaki, A M; Wimalasena, T T; Greetham, D

    2014-11-01

    Biofuels are expected to play a role in replacing crude oil as a liquid transportation fuel, and research into butanol has highlighted the importance of this alcohol as a fuel. Butanol has a higher energy density than ethanol, butanol-gasoline blends do not separate in the presence of water, and butanol is miscible with gasoline (Szulczyk, Int J Energy Environ 1(1):2876-2895, 40). Saccharomyces cerevisiae has been used as a fermentative organism in the biofuel industry producing ethanol from glucose derived from starchy plant material; however, it typically cannot tolerate butanol concentrations greater than 2 % (Luong, Biotechnol Bioeng 29 (2):242-248, 27). 90 Saccharomyces spp. strains were screened for tolerance to 1-butanol via a phenotypic microarray assay and we observed significant variation in response with the most tolerant strains (S. cerevisiae DBVPG1788, S. cerevisiae DBVPG6044 and S. cerevisiae YPS128) exhibiting tolerance to 4 % 1-butanol compared with S. uvarum and S. castelli strains, which were sensitive to 3 % 1-butanol. Response to butanol was confirmed using traditional yeast methodologies such as growth; it was observed that fermentations in the presence of butanol, when using strains with a tolerant background, were significantly faster. Assessing for genetic rationale for tolerance, it was observed that 1-butanol-tolerant strains, when compared with 1-butanol-sensitive strains, had an up-regulation of RPN4, a transcription factor which regulates proteasome genes. Analysing for the importance of RPN4, we observed that a Δrpn4 strain displayed a reduced rate of fermentation in the presence of 1-butanol when compared with the BY4741 background strain. This data will aid the development of breeding programmes to produce better strains for future bio-butanol production.

  9. Differential Antidepressant-Like Response to Lithium Treatment between Mouse Strains: Effects of Sex, Maternal Care, and Mixed Genetic Background

    PubMed Central

    Can, Adem; Piantadosi, Sean C.; Gould, Todd D.

    2013-01-01

    Background Lithium is a mood stabilizer with both antidepressant and antimanic properties, though its mechanism of action is unclear. Identifying the genetic factors that influence lithium's therapeutic actions will be an important step to assist in identifying such mechanisms. We previously reported that lithium treatment of male mice has antidepressant-like effects in the C57BL/6J strain but that such effects were absent in the BALB/cJ strain. Objectives To assess the roles of both genetic, and non-genetic factors such as sex and non-shared environmental factors that may mediate differential behavioral responses to lithium. Methods Mice were treated with lithium for ten days and then tested in the forced swim test followed by lithium discontinuation and retesting to assess effects of lithium withdrawal. We also assessed effects of sex and cross-fostering on lithium response between the C57BL/6J and BALB/cJ strains, and antidepressant-like effects of lithium in the hybrid CB6F1/J strain that is derived from C57BL/6J and BALB/cJ parental strains. Results Neither sex nor maternal care significantly influenced the differential antidepressant-like profile of lithium. Withdrawal from lithium treatment reversed antidepressant-like effects in the C57BL/6J strain, but had no effects in BALB/cJ mice. Lithium treatment did not result in antidepressant-like effects in the CB6F1/J strain. Conclusions Genetic factors are likely primarily responsible for differential antidepressant-like effects of lithium in the C57BL/6J and BALB/cJ strains. Future studies identifying such genetic factors may help to elucidate the neurobiological mechanisms of lithium's therapeutic actions. PMID:23503701

  10. Advanced solid elements for sheet metal forming simulation

    NASA Astrophysics Data System (ADS)

    Mataix, Vicente; Rossi, Riccardo; Oñate, Eugenio; Flores, Fernando G.

    2016-08-01

    The solid-shells are an attractive kind of element for the simulation of forming processes, due to the fact that any kind of generic 3D constitutive law can be employed without any additional hypothesis. The present work consists in the improvement of a triangular prism solid-shell originally developed by Flores[2, 3]. The solid-shell can be used in the analysis of thin/thick shell, undergoing large deformations. The element is formulated in total Lagrangian formulation, and employs the neighbour (adjacent) elements to perform a local patch to enrich the displacement field. In the original formulation a modified right Cauchy-Green deformation tensor (C) is obtained; in the present work a modified deformation gradient (F) is obtained, which allows to generalise the methodology and allows to employ the Pull-Back and Push-Forwards operations. The element is based in three modifications: (a) a classical assumed strain approach for transverse shear strains (b) an assumed strain approach for the in-plane components using information from neighbour elements and (c) an averaging of the volumetric strain over the element. The objective is to use this type of elements for the simulation of shells avoiding transverse shear locking, improving the membrane behaviour of the in-plane triangle and to handle quasi-incompressible materials or materials with isochoric plastic flow.

  11. Use of antisense RNA to modify the composition of cellulosomes produced by Clostridium cellulolyticum.

    PubMed

    Perret, Stéphanie; Maamar, Hédia; Bélaich, Jean-Pierre; Tardif, Chantal

    2004-01-01

    The enzymatic composition of the cellulosomes produced by Clostridium cellulolyticum was modified by inhibiting the synthesis of Cel48F that is the major cellulase of the cellulosomes. The strain ATCC 35319 (pSOSasrF) was developed to over-produce a 469 nucleotide-long antisense-RNA (asRNA) directed against the ribosome-binding site region and the beginning of the coding region of the cel48F mRNAs. The cellulolytic system secreted by the asRNA-producing strain showed a markedly lower amount of Cel48F, compared to the control strain transformed with the empty plasmid (pSOSzero). This was correlated with a 30% decrease of the specific activity of the cellulolytic system on Avicel cellulose, indicating that Cel48F plays an important role in the recalcitrant cellulose degradation. However, only minor effects were observed on the growth parameters on cellulose. In both transformant strains, cellulosome production was found to be reduced and two unknown proteins (P105 and P98) appeared as major components of their cellulolytic systems. These proteins did not contain any dockerin domain and were shown to be not included into the cellulosomes; they are expected to participate to the non-cellulosomal cellulolytic system of C. cellulolyticum.

  12. [Isolation of endophytic fungi from medicinal plant Brucea javanica and their microbial inhibition activity].

    PubMed

    Liang, Zi-Ning; Zhu, Hua; Lai, Kai-Ping; Chen, Long

    2014-04-01

    To isolate and identify endophytic fungi from Brucea javanica, and to detect the antimicrobial activity of these strains. Endophytic fungi were isolated by tissue inoculation culture and identified by conventional morphological characteristic method. Seven kinds of pathogenic fungi and three kinds of bacteria were used as targeting microbes to test microbial inhibition activities by agar plate antagonistic action and modified agar gel diffusion methods, respectively. A total of 83 endophytic fungi strains were isolated from the root, stem, leaf and fruit of Brucea javanica. 34 strains were obtained from the stem, 32 strains were obtained from the leaf, 15 strains were isolated from the root and 2 strains came from the fruit. These 73 strains which had been identified attribute to 5 orders, 6 families and 12 genera. For the isolated strains, 14 strains had antifungal activities against at least one pathogenic fungi, 9 strains showed antibacterial activities against one or more bacteria. Especially, the strain YJ-17 which belonged to Phomopsis genus showed the best inhibitory effect on the targeting microbes. The endophytic fungi from Brucea javanica show diversity and microbial inhibition activity, and are worthy for further study on plant disease controlling.

  13. Comparative Evaluation of Two Vaccine Candidates against Experimental Leishmaniasis Due to Leishmania major Infection in Four Inbred Mouse Strains▿

    PubMed Central

    Benhnini, Fouad; Chenik, Mehdi; Laouini, Dhafer; Louzir, Hechmi; Cazenave, Pierre André; Dellagi, Koussay

    2009-01-01

    Experimental leishmaniasis in BALB/c and C57BL/6 mice are the most investigated murine models that were used for the preclinical evaluation of Leishmania vaccine candidates. We have previously described two new inbred mouse strains named PWK and MAI issued from feral founders that also support the development of experimental leishmaniasis due to L. major. In this study, we sought to determine whether different mouse inbred strains generate concordant or discordant results when used to evaluate the potential of Leishmania proteins to protect against experimental leishmaniasis. To this end, two Leishmania proteins, namely, LACK (for Leishmania homolog of receptor for activated C kinase) and LmPDI (for L. major protein disulfide isomerase) were compared for their capacity to protect against experimental leishmaniasis in PWK, MAI, BALB/c, and C57BL/6 inbred mouse strains. Our data show that the capacity of Leishmania proteins to confer protection depends on the mouse strain used, stressing the important role played by the genetic background in shaping the immune response against the pathogen. These results may have important implications for the preclinical evaluation of candidate Leishmania vaccines: rather than using a single mouse strain, a panel of different inbred strains of various genetic backgrounds should be tested in parallel. The antigen that confers protection in the larger range of inbred strains may have better chances to be also protective in outbred human populations and should be selected for clinical trials. PMID:19726616

  14. Elongator complex influences telomeric gene silencing and DNA damage response by its role in wobble uridine tRNA modification.

    PubMed

    Chen, Changchun; Huang, Bo; Eliasson, Mattias; Rydén, Patrik; Byström, Anders S

    2011-09-01

    Elongator complex is required for formation of the side chains at position 5 of modified nucleosides 5-carbamoylmethyluridine (ncm⁵U₃₄), 5-methoxycarbonylmethyluridine (mcm⁵U₃₄), and 5-methoxycarbonylmethyl-2-thiouridine (mcm⁵s²U₃₄) at wobble position in tRNA. These modified nucleosides are important for efficient decoding during translation. In a recent publication, Elongator complex was implicated to participate in telomeric gene silencing and DNA damage response by interacting with proliferating cell nuclear antigen (PCNA). Here we show that elevated levels of tRNA(Lys)(s²UUU), tRNA(Gln)(s²UUG), and tRNA(Glu)(s²UUC), which in a wild-type background contain the mcm⁵s²U nucleoside at position 34, suppress the defects in telomeric gene silencing and DNA damage response observed in the Elongator mutants. We also found that the reported differences in telomeric gene silencing and DNA damage response of various elp3 alleles correlated with the levels of modified nucleosides at U₃₄. Defects in telomeric gene silencing and DNA damage response are also observed in strains with the tuc2Δ mutation, which abolish the formation of the 2-thio group of the mcm⁵s²U nucleoside in tRNA(Lys)(mcm⁵s²UUU), tRNA(Gln)(mcm⁵s²UUG), and tRNA(Glu)(mcm⁵s²UUC). These observations show that Elongator complex does not directly participate in telomeric gene silencing and DNA damage response, but rather that modified nucleosides at U₃₄ are important for efficient expression of gene products involved in these processes. Consistent with this notion, we found that expression of Sir4, a silent information regulator required for assembly of silent chromatin at telomeres, was decreased in the elp3Δ mutants.

  15. Elongator Complex Influences Telomeric Gene Silencing and DNA Damage Response by Its Role in Wobble Uridine tRNA Modification

    PubMed Central

    Chen, Changchun; Huang, Bo; Eliasson, Mattias; Rydén, Patrik; Byström, Anders S.

    2011-01-01

    Elongator complex is required for formation of the side chains at position 5 of modified nucleosides 5-carbamoylmethyluridine (ncm5U34), 5-methoxycarbonylmethyluridine (mcm5U34), and 5-methoxycarbonylmethyl-2-thiouridine (mcm5s2U34) at wobble position in tRNA. These modified nucleosides are important for efficient decoding during translation. In a recent publication, Elongator complex was implicated to participate in telomeric gene silencing and DNA damage response by interacting with proliferating cell nuclear antigen (PCNA). Here we show that elevated levels of tRNALys s2 UUU, tRNAGln s2 UUG, and tRNAGlu s2 UUC, which in a wild-type background contain the mcm5s2U nucleoside at position 34, suppress the defects in telomeric gene silencing and DNA damage response observed in the Elongator mutants. We also found that the reported differences in telomeric gene silencing and DNA damage response of various elp3 alleles correlated with the levels of modified nucleosides at U34. Defects in telomeric gene silencing and DNA damage response are also observed in strains with the tuc2Δ mutation, which abolish the formation of the 2-thio group of the mcm5s2U nucleoside in tRNALys mcm5s2UUU, tRNAGln mcm5s2UUG, and tRNAGlu mcm5s2UUC. These observations show that Elongator complex does not directly participate in telomeric gene silencing and DNA damage response, but rather that modified nucleosides at U34 are important for efficient expression of gene products involved in these processes. Consistent with this notion, we found that expression of Sir4, a silent information regulator required for assembly of silent chromatin at telomeres, was decreased in the elp3Δ mutants. PMID:21912530

  16. Weight gain in mice on a high caloric diet and chronically treated with omeprazole depends on sex and genetic background

    PubMed Central

    Tsao, Amy C.; Gillilland, Merritt G.; Merchant, Juanita L.

    2016-01-01

    The impact of omeprazole (OM), a widely used over-the-counter proton pump inhibitor, on weight gain has not been extensively explored. We examined what factors, e.g., diet composition, microbiota, genetic strain, and sex, might affect weight gain in mice fed a high caloric diet while on OM. Inbred C57BL/6J strain, a 50:50 hybrid (B6SJLF1/J) strain, and mice on a highly mixed genetic background were fed four diets: standard chow (STD, 6% fat), STD with 200 ppm OM (STD + O), a high-energy chow (HiE, 11% fat), and HiE chow with OM (HiE + O) for 17 wk. Metabolic analysis, body composition, and fecal microbiota composition were analyzed in C57BL/6J mice. Oral glucose tolerance tests were performed using mice on the mixed background. After 8 wk, female and male C57BL/6J mice on the HiE diets ate less, whereas males on the HiE diets compared with the STD diets gained weight. All diet treatments reduced energy expenditure in females but in males only those on the HiE + O diet. Gut microbiota composition differed in the C57BL/6J females but not the males. Hybrid B6SJLF1/J mice showed similar weight gain on all test diets. In contrast, mixed strain male mice fed a HiE + O diet gained ∼40% more weight than females on the same diet. In addition to increased weight gain, mixed genetic mice on the HiE + O diet cleared glucose normally but secreted more insulin. We concluded that sex and genetic background define weight gain and metabolic responses of mice on high caloric diets and OM. PMID:27810953

  17. Targeted Changes of the Cell Wall Proteome Influence Candida albicans Ability to Form Single- and Multi-strain Biofilms

    PubMed Central

    Walker, Louise A.; Martin-Yken, Hélène; Dague, Etienne; Legrand, Mélanie; Lee, Keunsook; Chauvel, Murielle; Firon, Arnaud; Rossignol, Tristan; Richard, Mathias L.; Munro, Carol A.; Bachellier-Bassi, Sophie; d'Enfert, Christophe

    2014-01-01

    Biofilm formation is an important virulence trait of the pathogenic yeast Candida albicans. We have combined gene overexpression, strain barcoding and microarray profiling to screen a library of 531 C. albicans conditional overexpression strains (∼10% of the genome) for genes affecting biofilm development in mixed-population experiments. The overexpression of 16 genes increased strain occupancy within a multi-strain biofilm, whereas overexpression of 4 genes decreased it. The set of 16 genes was significantly enriched for those encoding predicted glycosylphosphatidylinositol (GPI)-modified proteins, namely Ihd1/Pga36, Phr2, Pga15, Pga19, Pga22, Pga32, Pga37, Pga42 and Pga59; eight of which have been classified as pathogen-specific. Validation experiments using either individually- or competitively-grown overexpression strains revealed that the contribution of these genes to biofilm formation was variable and stage-specific. Deeper functional analysis of PGA59 and PGA22 at a single-cell resolution using atomic force microscopy showed that overexpression of either gene increased C. albicans ability to adhere to an abiotic substrate. However, unlike PGA59, PGA22 overexpression led to cell cluster formation that resulted in increased sensitivity to shear forces and decreased ability to form a single-strain biofilm. Within the multi-strain environment provided by the PGA22-non overexpressing cells, PGA22-overexpressing cells were protected from shear forces and fitter for biofilm development. Ultrastructural analysis, genome-wide transcript profiling and phenotypic analyses in a heterologous context suggested that PGA22 affects cell adherence through alteration of cell wall structure and/or function. Taken together, our findings reveal that several novel predicted GPI-modified proteins contribute to the cooperative behaviour between biofilm cells and are important participants during C. albicans biofilm formation. Moreover, they illustrate the power of using signature tagging in conjunction with gene overexpression for the identification of novel genes involved in processes pertaining to C. albicans virulence. PMID:25502890

  18. Viral proliferation and expression of tumor-related gene in different chicken embryo fibroblasts infected with different tumorigenic phenotypes of avian leukosis virus subgroup J.

    PubMed

    Qu, Yajin; Liu, Litao; Niu, Yujuan; Qu, Yue; Li, Ning; Sun, Wei; Lv, Chuanwei; Wang, Pengfei; Zhang, Guihua; Liu, Sidang

    2016-10-01

    Subgroup J avian leukosis virus (ALV-J) causes a neoplastic disease in infected chickens. The ALV-J strain NX0101, which was isolated from broiler breeders in 2001, mainly induced formation of myeloid cell tumors. However, strain HN10PY01, which was recently isolated from laying hens, mainly induces formation of myeloid cell tumors and hemangioma. To identify the molecular pathological mechanism underlying changes in host susceptibility and tumor classification induced by these two types of ALV-J strains, chicken embryo fibroblasts derived from chickens with different genetic backgrounds (broiler breeders and laying hens) and an immortalized chicken embryo fibroblasts (DF-1) were prepared and infected with strain NX0101 or HN10PY01, respectively. The 50% tissue culture infective dose (TCID50) and levels of ALV group-specific antigen p27 and heat shock protein 70 in the supernatant collected from the ALV-J infected cells were detected. Moreover, mRNA expression levels of tumor-related genes p53, c-myc, and Bcl-2 in ALV-J-infected cells were quantified. The results indicated that the infection of ALV-J could significantly increase mRNA expression levels of p53, c-myc, and Bcl-2 Strain HN10PY01 exhibited a greater influence on the three tumor-related genes in each of the three types of cells when compared with strain NX0101, and the TCID50 and p27 levels in the supernatant collected from HN10PY01-infected cells were higher than those collected from NX0101-infected cells. These results indicate that the infection of the two ALV-J strains influenced the gene expression levels in the infected cells, while the newly isolated strain HN10PY01 showed higher replication ability in cells and induced higher expression levels of tumor-related genes in infected cells. Furthermore, virus titers and expression levels of tumor-related genes and cellular stress responses of cells with different genetic backgrounds when infected with each of the two ALV-J strain were different, indicating that genetic backgrounds influenced the capabilities of the virus to infect and proliferate. The findings of this study provide useful data to further elucidate the mechanism underlying host susceptibility and tumor classification in ALV-J-infected chickens and cells. © 2016 Poultry Science Association Inc.

  19. A Single Multilocus Sequence Typing (MLST) Scheme for Seven Pathogenic Leptospira Species

    PubMed Central

    Amornchai, Premjit; Wuthiekanun, Vanaporn; Bailey, Mark S.; Holden, Matthew T. G.; Zhang, Cuicai; Jiang, Xiugao; Koizumi, Nobuo; Taylor, Kyle; Galloway, Renee; Hoffmaster, Alex R.; Craig, Scott; Smythe, Lee D.; Hartskeerl, Rudy A.; Day, Nicholas P.; Chantratita, Narisara; Feil, Edward J.; Aanensen, David M.; Spratt, Brian G.; Peacock, Sharon J.

    2013-01-01

    Background The available Leptospira multilocus sequence typing (MLST) scheme supported by a MLST website is limited to L. interrogans and L. kirschneri. Our aim was to broaden the utility of this scheme to incorporate a total of seven pathogenic species. Methodology and Findings We modified the existing scheme by replacing one of the seven MLST loci (fadD was changed to caiB), as the former gene did not appear to be present in some pathogenic species. Comparison of the original and modified schemes using data for L. interrogans and L. kirschneri demonstrated that the discriminatory power of the two schemes was not significantly different. The modified scheme was used to further characterize 325 isolates (L. alexanderi [n = 5], L. borgpetersenii [n = 34], L. interrogans [n = 222], L. kirschneri [n = 29], L. noguchii [n = 9], L. santarosai [n = 10], and L. weilii [n = 16]). Phylogenetic analysis using concatenated sequences of the 7 loci demonstrated that each species corresponded to a discrete clade, and that no strains were misclassified at the species level. Comparison between genotype and serovar was possible for 254 isolates. Of the 31 sequence types (STs) represented by at least two isolates, 18 STs included isolates assigned to two or three different serovars. Conversely, 14 serovars were identified that contained between 2 to 10 different STs. New observations were made on the global phylogeography of Leptospira spp., and the utility of MLST in making associations between human disease and specific maintenance hosts was demonstrated. Conclusion The new MLST scheme, supported by an updated MLST website, allows the characterization and species assignment of isolates of the seven major pathogenic species associated with leptospirosis. PMID:23359622

  20. The Effectiveness of Injury Prevention Programs to Modify Risk Factors for Non-Contact Anterior Cruciate Ligament and Hamstring Injuries in Uninjured Team Sports Athletes: A Systematic Review

    PubMed Central

    Monajati, Alireza; Larumbe-Zabala, Eneko; Goss-Sampson, Mark; Naclerio, Fernando

    2016-01-01

    Background Hamstring strain and anterior cruciate ligament injuries are, respectively, the most prevalent and serious non-contact occurring injuries in team sports. Specific biomechanical and neuromuscular variables have been used to estimate the risk of incurring a non-contact injury in athletes. Objective The aim of this study was to systematically review the evidences for the effectiveness of injury prevention protocols to modify biomechanical and neuromuscular anterior cruciate and/or hamstring injuries associated risk factors in uninjured team sport athletes. Data Sources PubMed, Science Direct, Web of Science, Cochrane Libraries, U.S. National Institutes of Health clinicaltrials.gov, Sport Discuss and Google Scholar databases were searched for relevant journal articles published until March 2015. A manual review of relevant articles, authors, and journals, including bibliographies was performed from identified articles. Main Results Nineteen studies were included in this review. Four assessment categories: i) landing, ii) side cutting, iii) stop-jump, and iv) muscle strength outcomes, were used to analyze the effectiveness of the preventive protocols. Eight studies using multifaceted interventions supported by video and/or technical feedback showed improvement in landing and/or stop-jump biomechanics, while no effects were observed on side-cutting maneuver. Additionally, multifaceted programs including hamstring eccentric exercises increased hamstring strength, hamstring to quadriceps functional ratio and/or promoted a shift of optimal knee flexion peak torque toward a more open angle position. Conclusions Multifaceted programs, supported by proper video and/or technical feedback, including eccentric hamstring exercises would positively modify the biomechanical and or neuromuscular anterior cruciate and/or hamstring injury risk factors. PMID:27171282

  1. Development and Characterization of an Infectious cDNA Clone of the Modified Live Virus Vaccine Strain of Equine Arteritis Virus

    PubMed Central

    Zhang, Jianqiang; Go, Yun Young; Huang, Chengjin M.; Meade, Barry J.; Lu, Zhengchun; Snijder, Eric J.; Timoney, Peter J.

    2012-01-01

    A stable full-length cDNA clone of the modified live virus (MLV) vaccine strain of equine arteritis virus (EAV) was developed. RNA transcripts generated from this plasmid (pEAVrMLV) were infectious upon transfection into mammalian cells, and the resultant recombinant virus (rMLV) had 100% nucleotide identity to the parental MLV vaccine strain of EAV. A single silent nucleotide substitution was introduced into the nucleocapsid gene (pEAVrMLVB), enabling the cloned vaccine virus (rMLVB) to be distinguished from parental MLV vaccine as well as other field and laboratory strains of EAV by using an allelic discrimination real-time reverse transcription (RT)-PCR assay. In vitro studies revealed that the cloned vaccine virus rMLVB and the parental MLV vaccine virus had identical growth kinetics and plaque morphologies in equine endothelial cells. In vivo studies confirmed that the cloned vaccine virus was very safe and induced high titers of neutralizing antibodies against EAV in experimentally immunized horses. When challenged with the heterologous EAV KY84 strain, the rMLVB vaccine virus protected immunized horses in regard to reducing the magnitude and duration of viremia and virus shedding but did not suppress the development of signs of EVA, although these were reduced in clinical severity. The vaccine clone pEAVrMLVB could be further manipulated to improve the vaccine efficacy as well as to develop a marker vaccine for serological differentiation of EAV naturally infected from vaccinated animals. PMID:22739697

  2. Country-level economic disparity and child mortality related to housing and injuries: a study in 26 European countries.

    PubMed

    Sengoelge, Mathilde; Elling, Berty; Laflamme, Lucie; Hasselberg, Marie

    2013-10-01

    Adverse living standards are associated with poorer child health and safety. This study investigates whether adverse housing and neighbourhood conditions contribute to explain country-level associations between a country's economic level and income inequality and child mortality, specifically injury mortality. Ecological, cross-sectional study. Twenty-six European countries were grouped according to two country-level economic measures from Eurostat: gross domestic product (GDP) and income inequality. Adverse country-level housing and neighbourhood conditions were assessed using data from the 2006 European Union Income Social Inclusion and Living Conditions Database (n=203 000). Child mortality incidence rates were derived for children aged 1-14 years for all causes, all injuries, road traffic injuries and unintentional injuries excluding road traffic. Linear regression analysis was applied to measure whether housing or neighbourhood conditions have a significant association with child mortality and whether a strain modified the association between GDP/income inequality and mortality. Country-level income inequality and GDP demonstrated a significant association with child mortality for all outcomes. A significant association was also found between housing strain and all child mortality outcomes, but not for neighbourhood strain. Housing strain partially modified the relationship between income inequality and GDP and all child mortality outcomes, with the exception of income inequality and road traffic injury mortality showing full mediation by housing strain. Adverse housing conditions are a likely pathway in the country-level association between income inequality and economic GDP and child injury mortality.

  3. Development of an Integrated Biofuel and Chemical Refinery

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

    Trawick, John; Burk, Mark; Barton, Nelson

    2017-02-06

    This project has demonstrated the level of commercial readiness for production of the industrial chemical, 1,4-butanediol (BDO), from lignocellulosic biomass by engineered E. coli. Targets were BDO titer, rate, and yield (TRY) and growth in lignocellulosic hydrolysates (Hz). A range of Hzs were used to assess limitations for biomass-to-BDO. Via adaptive evolution methods, whole-genome sequencing, and introduction of identified target genes, strains co-utilizing C5/ C6 sugars were made. The composition of Hz versus TRY led to a modified Hz composition. This was used in partnership with the DOE to redirect the project to focus on 1) several biomass Hz frommore » new suppliers, 2) Hz specification due to the characteristics of the Genomatica BDO process, 3) a gene cassette to engineer any BDO producing strain for biomass, and 4) modified BDO recovery to more economically recover BDO at industry specifications. BDO TRY and growth of the E. coli strains were predictable based on Hz composition from several suppliers. This defined metrics for biomass Hz composition to achieve BDO TRY along with internal TEA to evaluate the economic potential of each modification to strain, Hz feed, and process. An improved biomass-to-BDO production strain reached BDO T-R in a 30 L fermentation above original objectives. Yield approached the proposed Y and modifications to BDO recovery were demonstrated. Genomatica is now in the position of being able to incorporate biomass feedstocks into the commercial GENO BDO process.« less

  4. Pigs immunized with Chinese highly pathogenic PRRS virus modified live vaccine are protected from challenge with North American PRRSV strain NADC-20.

    PubMed

    Galliher-Beckley, Amy; Li, Xiangdong; Bates, John T; Madera, Rachel; Waters, Andrew; Nietfeld, Jerome; Henningson, Jamie; He, Dongsheng; Feng, Wenhai; Chen, Ruiai; Shi, Jishu

    2015-07-09

    Modified live virus (MLV) vaccines developed to protect against PRRSV circulating in North America (NA) offer limited protection to highly pathogenic (HP) PRRSV strains that are emerging in Asia. MLV vaccines specific to HP-PRRSV strains commercially available in China provide protection to HP-PRRSV; however, the efficacy of these HP-PRRSV vaccines to current circulating NA PRRS viruses has not been reported. The aim of this study is to investigate whether pigs vaccinated with attenuated Chinese HP-PRRSV vaccine (JXA1-R) are protected from infection by NA PRRSV strain NADC-20. We found that pigs vaccinated with JXA1-R were protected from challenges with HV-PRRSV or NADC-20 as shown by fewer days of clinical fever, reduced lung pathology scores, and lower PRRS virus load in the blood. PRRSV-specific antibodies, as measured by IDEXX ELISA, appeared one week after vaccination and virus neutralizing antibodies were detected four weeks post vaccination. Pigs vaccinated with JXA1-R developed broadly neutralizing antibodies with high titers to NADC-20, JXA1-R, and HV-PRRSV. In addition, we also found that IFN-α and IFN-β occurred at higher levels in the lungs of pigs vaccinated with JXA1-R. Taken together, our studies provide the first evidence that JXA1-R can confer protection in pigs against the heterologous NA PRRSV strain NADC-20. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  5. Bioethanol production from cellulosic hydrolysates by engineered industrial Saccharomyces cerevisiae.

    PubMed

    Lee, Ye-Gi; Jin, Yong-Su; Cha, Young-Lok; Seo, Jin-Ho

    2017-03-01

    Even though industrial yeast strains exhibit numerous advantageous traits for the production of bioethanol, their genetic manipulation has been limited. This study demonstrates that an industrial polyploidy Saccharomyces cerevisiae JHS200 can be engineered through Cas9 (CRISPR associated protein 9)-based genome editing. Specifically, we generated auxotrophic mutants and introduced a xylose metabolic pathway into the auxotrophic mutants. As expected, the engineered strain (JX123) enhanced ethanol production from cellulosic hydrolysates as compared to other engineered haploid strains. However, the JX123 strain produced substantial amounts of xylitol as a by-product during xylose fermentation. Hypothesizing that the xylitol accumulation might be caused by intracellular redox imbalance from cofactor difference, the NADH oxidase from Lactococcus lactis was introduced into the JX123 strain. The resulting strain (JX123_noxE) not only produced more ethanol, but also produced xylitol less than the JX123 strain. These results suggest that industrial polyploidy yeast can be modified for producing biofuels and chemicals. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Strain control of oxygen kinetics in the Ruddlesden-Popper oxide La 1.85Sr 0.15CuO 4

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

    Meyer, Tricia L.; Jacobs, Ryan; Lee, Dongkyu

    Oxygen defect control has long been considered an important route to functionalizing complex oxide films. However, the nature of oxygen defects in thin films is often not investigated beyond basic redox chemistry. One of the model examples for oxygen-defect studies is the layered Ruddlesden–Popper phase La 2-xSr x CuO 4-δ (LSCO), in which the superconducting transition temperature is highly sensitive to epitaxial strain. However, previous observations of strain-superconductivity coupling in LSCO thin films were mainly understood in terms of elastic contributions to mechanical buckling, with minimal consideration of kinetic or thermodynamic factors. Here, we report that the oxygen nonstoichiometry commonlymore » reported for strained cuprates is mediated by the strain-modified surface exchange kinetics, rather than reduced thermodynamic oxygen formation energies. Remarkably, tensile-strained LSCO shows nearly an order of magnitude faster oxygen exchange rate than a compressively strained film, providing a strategy for developing high-performance energy materials.« less

  7. Strain control of oxygen kinetics in the Ruddlesden-Popper oxide La 1.85Sr 0.15CuO 4

    DOE PAGES

    Meyer, Tricia L.; Jacobs, Ryan; Lee, Dongkyu; ...

    2018-01-08

    Oxygen defect control has long been considered an important route to functionalizing complex oxide films. However, the nature of oxygen defects in thin films is often not investigated beyond basic redox chemistry. One of the model examples for oxygen-defect studies is the layered Ruddlesden–Popper phase La 2-xSr x CuO 4-δ (LSCO), in which the superconducting transition temperature is highly sensitive to epitaxial strain. However, previous observations of strain-superconductivity coupling in LSCO thin films were mainly understood in terms of elastic contributions to mechanical buckling, with minimal consideration of kinetic or thermodynamic factors. Here, we report that the oxygen nonstoichiometry commonlymore » reported for strained cuprates is mediated by the strain-modified surface exchange kinetics, rather than reduced thermodynamic oxygen formation energies. Remarkably, tensile-strained LSCO shows nearly an order of magnitude faster oxygen exchange rate than a compressively strained film, providing a strategy for developing high-performance energy materials.« less

  8. Strain-engineered optoelectronic properties of 2D transition metal dichalcogenide lateral heterostructures

    DOE PAGES

    Lee, Jaekwang; Huang, Jingsong; Sumpter, Bobby G.; ...

    2017-02-17

    Compared with their bulk counterparts, 2D materials can sustain much higher elastic strain at which optical quantities such as bandgaps and absorption spectra governing optoelectronic device performance can be modified with relative ease. Using first-principles density functional theory and quasiparticle GW calculations, we demonstrate how uniaxial tensile strain can be utilized to optimize the electronic and optical properties of transition metal dichalcogenide lateral (in-plane) heterostructures such as MoX 2/WX 2 (X = S, Se, Te). We find that these lateral-type heterostructures may facilitate efficient electron–hole separation for light detection/harvesting and preserve their type II characteristic up to 12% of uniaxialmore » strain. Based on the strain-dependent bandgap and band offset, we show that uniaxial tensile strain can significantly increase the power conversion efficiency of these lateral heterostructures. Our results suggest that these strain-engineered lateral heterostructures are promising for optimizing optoelectronic device performance by selectively tuning the energetics of the bandgap.« less

  9. Suspension-Feeding Vertebrates.

    ERIC Educational Resources Information Center

    Sanderson, S. Laurie; Wassersug, Richard

    1990-01-01

    Described are animals that can filter their food out of the water. Various structures modified to strain plankton and small animals from water are detailed. The adaptive significance of these structures is discussed. (CW)

  10. Amplified fragment length polymorphism of Streptococcus suis strains correlates with their profile of virulence-associated genes and clinical background.

    PubMed

    Rehm, Thomas; Baums, Christoph G; Strommenger, Birgit; Beyerbach, Martin; Valentin-Weigand, Peter; Goethe, Ralph

    2007-01-01

    Amplified fragment length polymorphism (AFLP) typing was applied to 116 Streptococcus suis isolates with different clinical backgrounds (invasive/pneumonia/carrier/human) and with known profiles of virulence-associated genes (cps1, -2, -7 and -9, as well as mrp, epf and sly). A dendrogram was generated that allowed identification of two clusters (A and C) with different subclusters (A1, A2, C1 and C2) and two heterogeneous groups of strains (B and D). For comparison, three strains from each AFLP subcluster and group were subjected to multilocus sequence typing (MLST) analysis. The closest relationship and lowest diversity were found for patterns clustering within AFLP subcluster A1, which corresponded with sequence type (ST) complex 1. Strains within subcluster A1 were mainly invasive cps1 and mrp+ epf+ (or epf*) sly+ cps2+ strains of porcine or human origin. A new finding of this study was the clustering of invasive mrp* cps9 isolates within subcluster A2. MLST analysis suggested that A2 correlates with a single ST complex (ST87). In contrast to A1 and A2, subclusters C1 and C2 contained mainly pneumonia isolates of genotype cps7 or cps2 and epf- sly-. In conclusion, this study demonstrates that AFLP allows identification of clusters of S. suis strains with clinical relevance.

  11. Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus

    PubMed Central

    Yoshimi, Akira; Sano, Motoaki; Inaba, Azusa; Kokubun, Yuko; Fujioka, Tomonori; Mizutani, Osamu; Hagiwara, Daisuke; Fujikawa, Takashi; Nishimura, Marie; Yano, Shigekazu; Kasahara, Shin; Shimizu, Kiminori; Yamaguchi, Masashi; Kawakami, Kazuyoshi; Abe, Keietsu

    2013-01-01

    Although α-1,3-glucan is one of the major cell wall polysaccharides in filamentous fungi, the physiological roles of α-1,3-glucan remain unclear. The model fungus Aspergillus nidulans possesses two α-1,3-glucan synthase (AGS) genes, agsA and agsB. For functional analysis of these genes, we constructed several mutant strains in A. nidulans: agsA disruption, agsB disruption, and double-disruption strains. We also constructed several CagsB strains in which agsB expression was controlled by the inducible alcA promoter, with or without the agsA-disrupting mutation. The agsA disruption strains did not show markedly different phenotypes from those of the wild-type strain. The agsB disruption strains formed dispersed hyphal cells under liquid culture conditions, regardless of the agsA genetic background. Dispersed hyphal cells were also observed in liquid culture of the CagsB strains when agsB expression was repressed, whereas these strains grew normally in plate culture even under the agsB-repressed conditions. Fractionation of the cell wall based on the alkali solubility of its components, quantification of sugars, and 13C-NMR spectroscopic analysis revealed that α-1,3-glucan was the main component of the alkali-soluble fraction in the wild-type and agsA disruption strains, but almost no α-1,3-glucan was found in the alkali-soluble fraction derived from either the agsB disruption strain or the CagsB strain under the agsB-repressed conditions, regardless of the agsA genetic background. Taken together, our data demonstrate that the two AGS genes are dispensable in A. nidulans, but that AgsB is required for normal growth characteristics under liquid culture conditions and is the major AGS in this species. PMID:23365684

  12. Penicillin production in industrial strain Penicillium chrysogenum P2niaD18 is not dependent on the copy number of biosynthesis genes.

    PubMed

    Ziemons, Sandra; Koutsantas, Katerina; Becker, Kordula; Dahlmann, Tim; Kück, Ulrich

    2017-02-16

    Multi-copy gene integration into microbial genomes is a conventional tool for obtaining improved gene expression. For Penicillium chrysogenum, the fungal producer of the beta-lactam antibiotic penicillin, many production strains carry multiple copies of the penicillin biosynthesis gene cluster. This discovery led to the generally accepted view that high penicillin titers are the result of multiple copies of penicillin genes. Here we investigated strain P2niaD18, a production line that carries only two copies of the penicillin gene cluster. We performed pulsed-field gel electrophoresis (PFGE), quantitative qRT-PCR, and penicillin bioassays to investigate production, deletion and overexpression strains generated in the P. chrysogenum P2niaD18 background, in order to determine the copy number of the penicillin biosynthesis gene cluster, and study the expression of one penicillin biosynthesis gene, and the penicillin titer. Analysis of production and recombinant strain showed that the enhanced penicillin titer did not depend on the copy number of the penicillin gene cluster. Our assumption was strengthened by results with a penicillin null strain lacking pcbC encoding isopenicillin N synthase. Reintroduction of one or two copies of the cluster into the pcbC deletion strain restored transcriptional high expression of the pcbC gene, but recombinant strains showed no significantly different penicillin titer compared to parental strains. Here we present a molecular genetic analysis of production and recombinant strains in the P2niaD18 background carrying different copy numbers of the penicillin biosynthesis gene cluster. Our analysis shows that the enhanced penicillin titer does not strictly depend on the copy number of the cluster. Based on these overall findings, we hypothesize that instead, complex regulatory mechanisms are prominently implicated in increased penicillin biosynthesis in production strains.

  13. Tailoring automatic exposure control toward constant detectability in digital mammography.

    PubMed

    Salvagnini, Elena; Bosmans, Hilde; Struelens, Lara; Marshall, Nicholas W

    2015-07-01

    The automatic exposure control (AEC) modes of most full field digital mammography (FFDM) systems are set up to hold pixel value (PV) constant as breast thickness changes. This paper proposes an alternative AEC mode, set up to maintain some minimum detectability level, with the ultimate goal of improving object detectability at larger breast thicknesses. The default "opdose" AEC mode of a Siemens MAMMOMAT Inspiration FFDM system was assessed using poly(methyl methacrylate) (PMMA) of thickness 20, 30, 40, 50, 60, and 70 mm to find the tube voltage and anode/filter combination programmed for each thickness; these beam quality settings were used for the modified AEC mode. Detectability index (d'), in terms of a non-prewhitened model observer with eye filter, was then calculated as a function of tube current-time product (mAs) for each thickness. A modified AEC could then be designed in which detectability never fell below some minimum setting for any thickness in the operating range. In this study, the value was chosen such that the system met the achievable threshold gold thickness (Tt) in the European guidelines for the 0.1 mm diameter disc (i.e., Tt ≤ 1.10 μm gold). The default and modified AEC modes were compared in terms of contrast-detail performance (Tt), calculated detectability (d'), signal-difference-to-noise ratio (SDNR), and mean glandular dose (MGD). The influence of a structured background on object detectability for both AEC modes was examined using a CIRS BR3D phantom. Computer-based CDMAM reading was used for the homogeneous case, while the images with the BR3D background were scored by human observers. The default opdose AEC mode maintained PV constant as PMMA thickness increased, leading to a reduction in SDNR for the homogeneous background 39% and d' 37% in going from 20 to 70 mm; introduction of the structured BR3D plate changed these figures to 22% (SDNR) and 6% (d'), respectively. Threshold gold thickness (0.1 mm diameter disc) for the default AEC mode in the homogeneous background increased by 62% in going from 20 to 70 mm PMMA thickness; in the structured background, the increase was 39%. Implementation of the modified mode entailed an increase in mAs at PMMA thicknesses >40 mm; the modified AEC held threshold gold thickness constant above 40 mm PMMA with a maximum deviation of 5% in the homogeneous background and 3% in structured background. SDNR was also held constant with a maximum deviation of 4% and 2% for the homogeneous and the structured background, respectively. These results were obtained with an increase of MGD between 15% and 73% going from 40 to 70 mm PMMA thickness. This work has proposed and implemented a modified AEC mode, tailored toward constant detectability at larger breast thickness, i.e., above 40 mm PMMA equivalent. The desired improvement in object detectability could be obtained while maintaining MGD within the European guidelines achievable dose limit. (A study designed to verify the performance of the modified mode using more clinically realistic data is currently underway.).

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

    Salvagnini, Elena, E-mail: elena.salvagnini@uzleuven.be; Bosmans, Hilde; Struelens, Lara

    Purpose: The automatic exposure control (AEC) modes of most full field digital mammography (FFDM) systems are set up to hold pixel value (PV) constant as breast thickness changes. This paper proposes an alternative AEC mode, set up to maintain some minimum detectability level, with the ultimate goal of improving object detectability at larger breast thicknesses. Methods: The default “OPDOSE” AEC mode of a Siemens MAMMOMAT Inspiration FFDM system was assessed using poly(methyl methacrylate) (PMMA) of thickness 20, 30, 40, 50, 60, and 70 mm to find the tube voltage and anode/filter combination programmed for each thickness; these beam quality settingsmore » were used for the modified AEC mode. Detectability index (d′), in terms of a non-prewhitened model observer with eye filter, was then calculated as a function of tube current-time product (mAs) for each thickness. A modified AEC could then be designed in which detectability never fell below some minimum setting for any thickness in the operating range. In this study, the value was chosen such that the system met the achievable threshold gold thickness (T{sub t}) in the European guidelines for the 0.1 mm diameter disc (i.e., T{sub t} ≤ 1.10 μm gold). The default and modified AEC modes were compared in terms of contrast-detail performance (T{sub t}), calculated detectability (d′), signal-difference-to-noise ratio (SDNR), and mean glandular dose (MGD). The influence of a structured background on object detectability for both AEC modes was examined using a CIRS BR3D phantom. Computer-based CDMAM reading was used for the homogeneous case, while the images with the BR3D background were scored by human observers. Results: The default OPDOSE AEC mode maintained PV constant as PMMA thickness increased, leading to a reduction in SDNR for the homogeneous background 39% and d′ 37% in going from 20 to 70 mm; introduction of the structured BR3D plate changed these figures to 22% (SDNR) and 6% (d′), respectively. Threshold gold thickness (0.1 mm diameter disc) for the default AEC mode in the homogeneous background increased by 62% in going from 20 to 70 mm PMMA thickness; in the structured background, the increase was 39%. Implementation of the modified mode entailed an increase in mAs at PMMA thicknesses >40 mm; the modified AEC held threshold gold thickness constant above 40 mm PMMA with a maximum deviation of 5% in the homogeneous background and 3% in structured background. SDNR was also held constant with a maximum deviation of 4% and 2% for the homogeneous and the structured background, respectively. These results were obtained with an increase of MGD between 15% and 73% going from 40 to 70 mm PMMA thickness. Conclusions: This work has proposed and implemented a modified AEC mode, tailored toward constant detectability at larger breast thickness, i.e., above 40 mm PMMA equivalent. The desired improvement in object detectability could be obtained while maintaining MGD within the European guidelines achievable dose limit. (A study designed to verify the performance of the modified mode using more clinically realistic data is currently underway.)« less

  15. Efficacy of vaccination with La Sota strain vaccine to control Newcastle disease in village chickens in Nepal.

    PubMed

    Shrestha, Sulochana; Dhawan, Mamta; Donadeu, Meritxell; Dungu, Baptiste

    2017-02-01

    The efficacy of vaccination with Newcastle disease (ND) La Sota and R 2 B (Mukteswar) modified live strain vaccines was determined by experimental challenge and with ND La Sota vaccine under field conditions in Nepal. Booster vaccination with ND La Sota vaccine after a primary vaccination with ND La Sota vaccine, induced a geometric mean titre (GMT) of 5.0 log 2 haemagglutination inhibition (HI) units, compared to a GMT of 6.0 log 2 HI units following booster vaccination with R 2 B vaccine 1 month after primary vaccination with ND La Sota vaccine. Both vaccines provided 100% protection against challenge with a local field ND strain. Furthermore, booster vaccination with ND La Sota vaccine induced protective levels of antibody after field use in villages in Jhapa, and no outbreaks of ND occurred during the study period. The ND La Sota modified live vaccine is immunogenic and efficacious and is a suitable vaccine for use in vaccination programmes in village chickens in the rural areas of Nepal.

  16. A modified MacConkey agar for selective enumeration of necrotoxigenic E. coli O55 and probiotic E. coli Nissle 1917.

    PubMed

    Splichalova, Alla; Splichal, Igor; Sonnenborn, Ulrich; Rada, Vojtech

    2014-09-01

    An agar selective enumeration of necrotoxigenic Escherichia coli O55 (NTEC2) and probiotic E. coli Nissle 1917, using modified MacConkey agar, was developed to study bacterial interference between these E. coli strains in a gnotobiotic piglet model. Replacement of lactose with saccharose in the agar enables the direct visual enumeration of red colonies of E. coli O55 and yellow colonies of E. coli Nissle 1917 that are co-cultured in the same Petri dish. A total of 336 colonies (168 for each color) were subjected to strain-specific PCR identification with LNA probes. Sensitivity, specificity, and positive and negative predictive values were 96.43%, 95.83%, 95.86% and 96.41% respectively in E. coli O55, and 98.21%, 97.02%, 97.06% and 98.19% respectively in E. coli Nissle 1917. Color-based enumeration of both E. coli strains in colonic contents and mesenteric lymph nodes homogenates of gnotobiotic piglets demonstrated the applicability of this method for the gnotobiotic piglet model of enteric diseases. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Size–strain separation in diffraction line profile analysis

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

    Scardi, P.; Ermrich, M.; Fitch, A.

    Separation of size and strain effects on diffraction line profiles has been studied in a round robin involving laboratory instruments and synchrotron radiation beamlines operating with different radiation, optics, detectors and experimental configurations. The studied sample, an extensively ball milled iron alloy powder, provides an ideal test case, as domain size broadening and strain broadening are of comparable size. The high energy available at some synchrotron radiation beamlines provides the best conditions for an accurate analysis of the line profiles, as the size–strain separation clearly benefits from a large number of Bragg peaks in the pattern; high counts, reliable intensitymore » values in low-absorption conditions, smooth background and data collection at different temperatures also support the possibility to include diffuse scattering in the analysis, for the most reliable assessment of the line broadening effect. However, results of the round robin show that good quality information on domain size distribution and microstrain can also be obtained using standard laboratory equipment, even when patterns include relatively few Bragg peaks, provided that the data are of good quality in terms of high counts and low and smooth background.« less

  18. Size–strain separation in diffraction line profile analysis

    DOE PAGES

    Scardi, P.; Ermrich, M.; Fitch, A.; ...

    2018-05-29

    Separation of size and strain effects on diffraction line profiles has been studied in a round robin involving laboratory instruments and synchrotron radiation beamlines operating with different radiation, optics, detectors and experimental configurations. The studied sample, an extensively ball milled iron alloy powder, provides an ideal test case, as domain size broadening and strain broadening are of comparable size. The high energy available at some synchrotron radiation beamlines provides the best conditions for an accurate analysis of the line profiles, as the size–strain separation clearly benefits from a large number of Bragg peaks in the pattern; high counts, reliable intensitymore » values in low-absorption conditions, smooth background and data collection at different temperatures also support the possibility to include diffuse scattering in the analysis, for the most reliable assessment of the line broadening effect. However, results of the round robin show that good quality information on domain size distribution and microstrain can also be obtained using standard laboratory equipment, even when patterns include relatively few Bragg peaks, provided that the data are of good quality in terms of high counts and low and smooth background.« less

  19. Determination of atomic vacancies in InAs/GaSb strained-layer superlattices by atomic strain

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

    Kim, Honggyu; Meng, Yifei; Kwon, Ji-Hwan

    Determining vacancy in complex crystals or nanostructures represents an outstanding crystallographic problem that has a large impact on technology, especially for semiconductors, where vacancies introduce defect levels and modify the electronic structure. However, vacancy is hard to locate and its structure is difficult to probe experimentally. Reported here are atomic vacancies in the InAs/GaSb strained-layer superlattice (SLS) determined by atomic-resolution strain mapping at picometre precision. It is shown that cation and anion vacancies in the InAs/GaSb SLS give rise to local lattice relaxations, especially the nearest atoms, which can be detected using a statistical method and confirmed by simulation. Themore » ability to map vacancy defect-induced strain and identify its location represents significant progress in the study of vacancy defects in compound semiconductors.« less

  20. Biological indicators for low temperature steam and formaldehyde sterilization: investigation of the effect of change in temperature and formaldehyde concentration on spores of Bacillus stearothermophilus NCIMB 8224.

    PubMed

    Wright, A M; Hoxey, E V; Soper, C J; Davies, D J

    1996-03-01

    Five strains of Bacillus stearothermophilus have been studied to identify a spore strain to be used as a biological indicator organism for low temperature steam and formaldehyde sterilization. Three strains gave poor reproducibility of batch size and growth index and were discarded. The other two strains gave good reproducibility with a high growth index and gave rise to linear survivor curves when exposed to 5% aqueous formaldehyde. However, only NCIMB 8224 sporulates on a simpler medium and as it was the most resistant to formaldehyde, it was further studied. Tests were carried out in a modified miniclave and factors studied included temperature of the steam and formaldehyde concentration. All studies confirmed the suitability of this strain as a biological indicator organism.

  1. Determination of atomic vacancies in InAs/GaSb strained-layer superlattices by atomic strain

    DOE PAGES

    Kim, Honggyu; Meng, Yifei; Kwon, Ji-Hwan; ...

    2018-01-01

    Determining vacancy in complex crystals or nanostructures represents an outstanding crystallographic problem that has a large impact on technology, especially for semiconductors, where vacancies introduce defect levels and modify the electronic structure. However, vacancy is hard to locate and its structure is difficult to probe experimentally. Reported here are atomic vacancies in the InAs/GaSb strained-layer superlattice (SLS) determined by atomic-resolution strain mapping at picometre precision. It is shown that cation and anion vacancies in the InAs/GaSb SLS give rise to local lattice relaxations, especially the nearest atoms, which can be detected using a statistical method and confirmed by simulation. Themore » ability to map vacancy defect-induced strain and identify its location represents significant progress in the study of vacancy defects in compound semiconductors.« less

  2. [A structural protein study of the influenza A (H1N1) virus by polyacrylamide gel electrophoresis].

    PubMed

    Pérez Guevara, M T; Savón Valdés, C; Rivas Arjona, M; Goyenechea Hernández, A

    1992-01-01

    Influenza is an acute respiratory disease typically appearing as an epidemic. Three immunological types of the influenza virus are known: A, B and C. Continually, antigen changes occur, especially in type A. Therefore, a comparative study was carried out on 4 influenza A(H1N1) virus strains in relation to protein structure (surface antigens), by using polyacrylamide gel electrophoresis by the modified Laemmli method. The objective was to compare the structural proteins of the A/Havana/1292/78 (H1N1) national strain with the proteins of 3 international pattern strains. In all the cases, 6 bands were detected by densitometry. In the 4 strains studied the most abundant protein was M. Great differences between the Cuban strain and the 3 international patterns were not seen.

  3. Omics Analyses of Trichoderma reesei CBS999.97 and QM6a Indicate the Relevance of Female Fertility to Carbohydrate-Active Enzyme and Transporter Levels.

    PubMed

    Tisch, Doris; Pomraning, Kyle R; Collett, James R; Freitag, Michael; Baker, Scott E; Chen, Chia-Ling; Hsu, Paul Wei-Che; Chuang, Yu Chien; Schuster, Andre; Dattenböck, Christoph; Stappler, Eva; Sulyok, Michael; Böhmdorfer, Stefan; Oberlerchner, Josua; Wang, Ting-Fang; Schmoll, Monika

    2017-11-15

    The filamentous fungus Trichoderma reesei is found predominantly in the tropics but also in more temperate regions, such as Europe, and is widely known as a producer of large amounts of plant cell wall-degrading enzymes. We sequenced the genome of the sexually competent isolate CBS999.97, which is phenotypically different from the female sterile strain QM6a but can cross sexually with QM6a. Transcriptome data for growth on cellulose showed that entire carbohydrate-active enzyme (CAZyme) families are consistently differentially regulated between these strains. We evaluated backcrossed strains of both mating types, which acquired female fertility from CBS999.97 but maintained a mostly QM6a genetic background, and we could thereby distinguish between the effects of strain background and female fertility or mating type. We found clear regulatory differences associated with female fertility and female sterility, including regulation of CAZyme and transporter genes. Analysis of carbon source utilization, transcriptomes, and secondary metabolites in these strains revealed that only a few changes in gene regulation are consistently correlated with different mating types. Different strain backgrounds (QM6a versus CBS999.97) resulted in the most significant alterations in the transcriptomes and in carbon source utilization, with decreased growth of CBS999.97 on several amino acids (for example proline or alanine), which further correlated with the downregulation of genes involved in the respective pathways. In combination, our findings support a role of fertility-associated processes in physiology and gene regulation and are of high relevance for the use of sexual crossing in combining the characteristics of two compatible strains or quantitative trait locus (QTL) analysis. IMPORTANCE Trichoderma reesei is a filamentous fungus with a high potential for secretion of plant cell wall-degrading enzymes. We sequenced the genome of the fully fertile field isolate CBS999.97 and analyzed its gene regulation characteristics in comparison with the commonly used laboratory wild-type strain QM6a, which is not female fertile. Additionally, we also evaluated fully fertile strains with genotypes very close to that of QM6a in order to distinguish between strain-specific and fertility-specific characteristics. We found that QM6a and CBS999.97 clearly differ in their growth patterns on different carbon sources, CAZyme gene regulation, and secondary metabolism. Importantly, we found altered regulation of 90 genes associated with female fertility, including CAZyme genes and transporter genes, but only minor mating type-dependent differences. Hence, when using sexual crossing in research and for strain improvement, it is important to consider female fertile and female sterile strains for comparison with QM6a and to achieve optimal performance. Copyright © 2017 Tisch et al.

  4. Investigation of the Effect of Oil Modification on Critical Characteristics of Asphalt Binders

    NASA Astrophysics Data System (ADS)

    Golalipour, Amir

    Thermally induced cracking of asphalt pavement continues to be a serious issue in cold climate regions as well as in areas which experience extreme daily temperature differentials. Low temperature cracking of asphalt pavements is attributed to thermal stresses and strains developed during cooling cycles. Improving asphalt binder low temperature fracture and stiffness properties continues to be a subject of particular concern. Therefore, significant amount of research has been focused on improving asphalt binder properties through modification. In recent years, wide ranges of oil based modifications have been introduced to improve asphalt binder performance, especially at the low service temperatures. Although, significant use of these oils is seen in practice, knowledge of the fundamental mechanisms of oil modification and their properties for achieving optimum characteristics is limited. Hence, this study focuses on better understanding of the effect of oil modifiers which would help better material selection and achieve optimum performance in terms of increasing the life span of pavements. In this study, the effect of oil modification on the rheological properties of the asphalt binder is investigated. To examine the effect of oil modification on binder characteristics, low temperature properties as well as high temperature performance of oil modified binders were evaluated. It is found that oils vary in their effects on asphalt binder performance. However, for all oils used in the study, adding an oil to binder can improve binder low temperature performance, and this result mainly attributed to the softening effect. In addition to that, a simple linear model is proposed to predict the performance grade of oil modified binder based on the properties of its constituents at high and low temperatures. Another part of this study focuses on the oil modification effect on asphalt binder thermal strain and stresses. A viscoelastic analytical procedure is combined with experimentally derived failure stress and strain envelopes to determine the controlling failure mechanism, strain tolerance or critical stress, in thermal cracking of oil modified binders. The low temperature failure results depict that oil modification has a good potential of improving the cracking resistance of asphalt binders during thermal cycles.

  5. Strain gage measurement errors in the transient heating of structural components

    NASA Technical Reports Server (NTRS)

    Richards, W. Lance

    1993-01-01

    Significant strain-gage errors may exist in measurements acquired in transient thermal environments if conventional correction methods are applied. Conventional correction theory was modified and a new experimental method was developed to correct indicated strain data for errors created in radiant heating environments ranging from 0.6 C/sec (1 F/sec) to over 56 C/sec (100 F/sec). In some cases the new and conventional methods differed by as much as 30 percent. Experimental and analytical results were compared to demonstrate the new technique. For heating conditions greater than 6 C/sec (10 F/sec), the indicated strain data corrected with the developed technique compared much better to analysis than the same data corrected with the conventional technique.

  6. A New Correction Technique for Strain-Gage Measurements Acquired in Transient-Temperature Environments

    NASA Technical Reports Server (NTRS)

    Richards, W. Lance

    1996-01-01

    Significant strain-gage errors may exist in measurements acquired in transient-temperature environments if conventional correction methods are applied. As heating or cooling rates increase, temperature gradients between the strain-gage sensor and substrate surface increase proportionally. These temperature gradients introduce strain-measurement errors that are currently neglected in both conventional strain-correction theory and practice. Therefore, the conventional correction theory has been modified to account for these errors. A new experimental method has been developed to correct strain-gage measurements acquired in environments experiencing significant temperature transients. The new correction technique has been demonstrated through a series of tests in which strain measurements were acquired for temperature-rise rates ranging from 1 to greater than 100 degrees F/sec. Strain-gage data from these tests have been corrected with both the new and conventional methods and then compared with an analysis. Results show that, for temperature-rise rates greater than 10 degrees F/sec, the strain measurements corrected with the conventional technique produced strain errors that deviated from analysis by as much as 45 percent, whereas results corrected with the new technique were in good agreement with analytical results.

  7. Nest covering in plovers: How modifying the visual environment influences egg camouflage.

    PubMed

    Troscianko, Jolyon; Wilson-Aggarwal, Jared; Spottiswoode, Claire N; Stevens, Martin

    2016-10-01

    Camouflage is one of the most widespread antipredator defences, and its mechanistic basis has attracted considerable interest in recent years. The effectiveness of camouflage depends on the interaction between an animal's appearance and its background. Concealment can therefore be improved by changes to an animal's own appearance, by behaviorally selecting an optimal background, or by modifying the background to better match the animal's own appearance. Research to date has largely focussed on the first of these mechanisms, whereas there has been little work on the second and almost none on the third. Even though a number of animal species may potentially modify their environment to improve individual-specific camouflage, this has rarely if ever been quantitatively investigated, or its adaptive value tested. Kittlitz's plovers (Charadrius pecuarius) use material (stones and vegetation) to cover their nests when predators approach, providing concealment that is independent of the inflexible appearance of the adult or eggs, and that can be adjusted to suit the local surrounding background. We used digital imaging and predator vision modeling to investigate the camouflage properties of covered nests, and whether their camouflage affected their survival. The plovers' nest-covering materials were consistent with a trade-off between selecting materials that matched the color of the eggs, while resulting in poorer nest pattern and contrast matching to the nest surroundings. Alternatively, the systematic use of materials with high-contrast and small-pattern grain sizes could reflect a deliberate disruptive coloration strategy, whereby high-contrast material breaks up the telltale outline of the clutch. No camouflage variables predicted nest survival. Our study highlights the potential for camouflage to be enhanced by background modification. This provides a flexible system for modifying an animal's conspicuousness, to which the main limitation may be the available materials rather than the animal's appearance.

  8. A kinematically driven anisotropic viscoelastic constitutive model applied to tires

    NASA Technical Reports Server (NTRS)

    Johnson, Arthur R.; Tanner, John A.; Mason, Angela J.

    1995-01-01

    Aircraft tires are composite structures manufactured with viscoelastic materials such as carbon black filled rubber and nylon cords. When loaded they experience large deflections and moderately large strains. Detailed structural models of tires require the use of either nonlinear shell or nonlinear three dimensional solid finite elements. Computational predictions of the dynamic response of tires must consider the composite viscoelastic material behavior in a realistic fashion. We describe a modification to a nonlinear anisotropic shell finite element so it can be used to model viscoelastic stresses during general deformations. The model is developed by introducing internal variables of the type used to model elastic strain energy. The internal variables are strains, curvatures, and transverse shear angles which are in a one-to-one correspondence with the generalized coordinates used to model the elastic strain energy for nonlinear response. A difference-relaxation equation is used to relate changes in the observable strain field to changes in the internal strain field. The internal stress state is introduced into the equilibrium equations by converting it to nodal loads associated with the element's displacement degrees of freedom. In this form the tangent matrix in the Newton-Raphson solution algorithm is not modified from its form for the nonlinear statics problem. Only the gradient vector is modified and the modification is not computationally costly. The existing finite element model for the Space Shuttle nose gear tire is used to provide examples of the algorithm. In the first example, the tire's rim is displaced at a constant rate up to a fixed value. In the second example, the tire's rim is enforced to follow a saw tooth load and unload curve to generate hysteresis loops.

  9. A kinematically driven anisotropic viscoelastic constitutive model applied to tires

    NASA Astrophysics Data System (ADS)

    Johnson, Arthur R.; Tanner, John A.; Mason, Angela J.

    1995-08-01

    Aircraft tires are composite structures manufactured with viscoelastic materials such as carbon black filled rubber and nylon cords. When loaded they experience large deflections and moderately large strains. Detailed structural models of tires require the use of either nonlinear shell or nonlinear three dimensional solid finite elements. Computational predictions of the dynamic response of tires must consider the composite viscoelastic material behavior in a realistic fashion. We describe a modification to a nonlinear anisotropic shell finite element so it can be used to model viscoelastic stresses during general deformations. The model is developed by introducing internal variables of the type used to model elastic strain energy. The internal variables are strains, curvatures, and transverse shear angles which are in a one-to-one correspondence with the generalized coordinates used to model the elastic strain energy for nonlinear response. A difference-relaxation equation is used to relate changes in the observable strain field to changes in the internal strain field. The internal stress state is introduced into the equilibrium equations by converting it to nodal loads associated with the element's displacement degrees of freedom. In this form the tangent matrix in the Newton-Raphson solution algorithm is not modified from its form for the nonlinear statics problem. Only the gradient vector is modified and the modification is not computationally costly. The existing finite element model for the Space Shuttle nose gear tire is used to provide examples of the algorithm. In the first example, the tire's rim is displaced at a constant rate up to a fixed value. In the second example, the tire's rim is enforced to follow a saw tooth load and unload curve to generate hysteresis loops.

  10. Regulation of osteoarthritis by omega-3 (n-3) polyunsaturated fatty acids in a naturally occurring model of disease

    PubMed Central

    Knott, L.; Avery, N.C.; Hollander, A.P.; Tarlton, J.F.

    2011-01-01

    Summary Objective To examine effects of high omega-3 (n-3) polyunsaturated fatty acid (PUFA) diets on development of osteoarthritis (OA) in a spontaneous guinea pig model, and to further characterise pathogenesis in this model. Modern diets low in n-3 PUFAs have been linked with increases in inflammatory disorders, possibly including OA. However, n-3 is also thought to increases bone density, which is a possible contributing factor in OA. Therefore we aim to determine the net influence of n-3 in disease development. Method OA-prone Dunkin-Hartley (DH) Guinea pigs were compared with OA-resistant Bristol Strain-2s (BS2) each fed a standard or an n-3 diet from 10 to 30 weeks (10/group). We examined cartilage and subchondral bone pathology by histology, and biochemistry, including collagen cross-links, matrix metalloproteinases (MMPs), alkaline phosphatase, glycosaminoglycan (GAG), and denatured type II collagen. Results Dietary n-3 reduced disease in OA-prone animals. Most cartilage parameters were modified by n-3 diet towards those seen in the non-pathological BS2 strain – significantly active MMP-2, lysyl-pyridinoline and total collagen cross-links – the only exception being pro MMP-9 which was lower in the BS2, yet increased with n-3. GAG content was higher and denatured type II lower in the n-3 group. Subchondral bone parameters in the DH n-3 group also changed towards those seen in the non-pathological strain, significantly calcium:phosphate ratios and epiphyseal bone density. Conclusion Dietary n-3 PUFA reduced OA in the prone strain, and most disease markers were modified towards those of the non-OA strain, though not all significantly so. Omega-3 did not increase markers of pathology in either strain. PMID:21723952

  11. The evolution of γ-Mg17Al12 intermetallic compound during accumulative back extrusion and subsequent ageing treatment

    NASA Astrophysics Data System (ADS)

    Maghsoudi, M. H.; Zarei-Hanzaki, A.; Abedi, H. R.; Shamsolhodaei, A.

    2015-11-01

    Accumulative back extrusion (ABE) processing, as a novel severe plastic deformation (SPD) method, has been recently justified to be capable of modifying the microstructural characteristics of alloys. In line to its ongoing researches, the present work has been planned to study the evolution of γ-Mg17Al12 intermetallic phase during ABE and subsequent ageing treatment in a high Al-bearing Mg-Al-Zn alloy. The behaviour of γ intermetallic has been systematically examined as following points of view: (i) strain-temperature-dependent morphology changes, (ii) strain-induced dissolution, and (iii) re-ageing behaviour as a function of time and temperature. Aiming to analyse the morphology of eutectic γ compound with respect to the strain and temperature, 2D projections of effective diameter, shape factor and globularity have been made in strain/temperature graphs. The processing conditions (strain and temperature) corresponding to the desired and undesired morphologies are introduced and microstructurally explained through underlying plasticity mechanisms, i.e., 'necking-thinning-particle separation' and 'brittle fragmentation.' The former mechanism is suggested to be in relation with partial strain-induced dissolution of eutectic γ phase, leading to generation of a supersaturated solid solution. This has resulted to the observation of 'off-stoichiometry' phenomena in Mg17Al12 phase and has been justified through dislocation-assisted deformation mechanism at elevated temperature. Surprisingly, a unique re-ageing behaviour has been found for the obtained solid solutions, where a modified kinetics and morphology of γ phase precipitation were characterized. The altered precipitation behaviour is attributed to the specific defect structure achieved by SPD acting as fast diffusion channel for Al solutes.

  12. Alcohol dehydrogenase AdhA plays a role in ethanol tolerance in model cyanobacterium Synechocystis sp. PCC 6803.

    PubMed

    Vidal, Rebeca

    2017-04-01

    The protein AdhA from the cyanobacterium Synechocystis sp. PCC 6803 (hereafter Synechocystis) has been previously reported to show alcohol dehydrogenase activity towards ethanol and both NAD and NADP. This protein is currently being used in genetically modified strains of Synechocystis capable of synthesizing ethanol showing the highest ethanol productivities. In the present work, mutant strains of Synechocystis lacking AdhA have been constructed and tested for tolerance to ethanol. The lack of AdhA in the wild-type strain reduces survival to externally added ethanol at lethal concentration of 4% (v/v). On the other hand, the lack of AdhA in an ethanologenic strain diminishes tolerance of cells to internally produced ethanol. It is also shown that light-activated heterotrophic growth (LAHG) of the wild-type strain is impaired in the mutant strain lacking AdhA (∆adhA strain). Photoautotrophic, mixotrophic, and photoheterotrophic growth are not affected in the mutant strain. Based on phenotypic characterization of ∆adhA mutants, the possible physiological function of AdhA in Synechocystis is discussed.

  13. Detection of cholera (ctx) and zonula occludens (zot) toxin genes in Vibrio cholerae O1, O139 and non-O1 strains.

    PubMed

    Rivera, I G; Chowdhury, M A; Sanchez, P S; Sato, M I; Huq, A; Colwell, R R; Martins, M T

    1995-09-01

    Vibrio cholerae O1 and V. cholerae non-O1 strains isolated from environmental samples collected in São Paulo, Brazil, during cholera epidemics and pre-epidemic periods were examined for the presence of toxin genes. V. cholerae O1 strains isolated from clinical samples in Peru and Mexico, and V. cholerae O139 strains from India were also examined for the presence of ctx (cholera toxin gene) and zot (zonula occludens toxin gene) by polymerase chain reaction (PCR). A modified DNA-extraction method applied in this study yielded satisfactory recovery of genomic DNA from vibrios. Results showed that strains of V. cholerae O1 isolated during the preepidemic period were ctx (-)/zot (-) whereas strains isolated during the epidemic were ctx (+)/zot (+). All V. cholerae non-O1 strains tested in the study were ctx (-)/zot (-), whereas all V. cholerae O139 strains were ctx (+)/zot (+). Rapid detection of the virulence genes (ctx and zot) can be achieved by PCR and this can serve as an important tool in the epidemiology and surveillance of V. cholerae.

  14. 7 CFR 331.3 - PPQ select agents and toxins.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...) Select agents and toxins listed in paragraph (b) of this section that have been genetically modified. (d... variegated chlorosis strain). (c) Genetic elements, recombinant nucleic acids, and recombinant organisms: (1...

  15. 7 CFR 331.3 - PPQ select agents and toxins.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...) Select agents and toxins listed in paragraph (b) of this section that have been genetically modified. (d... variegated chlorosis strain). (c) Genetic elements, recombinant nucleic acids, and recombinant organisms: (1...

  16. Production of Antilisterial Bacteriocins from Lactic Acid Bacteria in Dairy-Based Media: A Comparative Study.

    PubMed

    Ünlü, Gülhan; Nielsen, Barbara; Ionita, Claudia

    2015-12-01

    One hundred and eight strains of lactic acid bacteria (LAB) were screened for bacteriocin production by the modified deferred antagonism and agar well diffusion methods. When the modified deferred antagonism method was employed, 82 LAB strains showed inhibitory action against Listeria monocytogenes v7 ½a, whereas 26 LAB strains expressed no inhibition. Only 12 LAB strains exhibited inhibitory activity when the agar well diffusion method was used, 11 of which had been previously recognized as bacteriocin production positive (Bac(+)). Lactobacillus viridescens NRRL B-1951 was determined, for the first time, to produce an inhibitory compound with a proteinaceous nature. The inhibitory activity was observed in the presence of lipase, α-chymotrypsin, and trypsin, but no inhibition zone could be detected in the presence of proteinase K, indicating the proteinaceous nature of the inhibitory compound. The inhibitory compound was active against Lact. sake ATCC 15521 and Lact. plantarum NCDO 995. Bacteriocin production by the Bac(+) LAB strains was assessed in Lactobacillus MRS Broth as well as in dairy-based media such as nonfat milk, demineralized whey powder, and cheddar cheese whey supplemented with complex nutrient sources that are rich in nitrogen. Lact. sake ATCC 15521 and L. monocytogenes CWD 1002, CWD 1092, CWD 1157, CWD 1198, and v7 ½a were used as indicators. The inhibitory activities of the bacteriocins varied depending on the indicator strains and the growth media used. The LAB indicator strains were found to be more sensitive to inhibition by bacteriocins when compared to the listerial indicator strains. Among the listerial indicators, L. monocytogenes CWD 1002 and CWD 1198 were the most sensitive strains to the bacteriocins investigated in this study. Media composition had a significant influence on bacteriocin production and activity. When compared to demineralized whey powder medium and cheddar cheese whey medium supplemented with whey protein concentrate, cheddar cheese whey medium supplemented with complex nutrient sources such as yeast extract, polypeptone, proteose peptone nr. 3, or soytone appeared to be more supportive of bacteriocin production.

  17. Compatibility of a wild type and its genetically modified Sinorhizobium strain with two mycorrhizal fungi on Medicago species as affected by drought stress.

    PubMed

    Vázquez, M M.; Azcón, R; Barea, J M.

    2001-07-01

    The effect of double inoculation with two strains of Sinorhizobium meliloti [the wild type (WT) strain GR4 and its genetically modified (GM) derivative GR4(pCK3)], and two species of arbuscular mycorrhizal (AM) fungi (Glomus deserticola and Glomus intraradices) was examined in a microcosm system on three species of Medicago (M. nolana, M. rigidula, M. rotata). Two water regimes (80 and 100% water holding capacity, WHC) were assayed. The efficiency of each AM fungus increasing plant growth, nutrient content, nodulation and water-stress tolerance was related to the Sinorhizobium strains and Medicago species. This indicates selective and specific compatibilities between microsymbionts and the common host plant. Differential effects of the mycorrhizal isolates were not associated with their colonizing ability. Nodulation and mycorrhizal dependency (MD) changed in each plant genotype in accordance with the Sinorhizobium strain and AM fungi involved. Generally, Medicago sp. MD decreased under water-stress conditions even when these conditions did not affect AM colonization (%). Proline accumulation in non-mycorrhizal plant leaves was increased by water stress, except in M. rotata plants. Differences in proline accumulation in AM-colonized plants suggest that both the AM fungus and the Sinorhizobium strain were able to induce different degrees of osmotic adjustment. Mycorrhizal plants nodulated by the WT strain accumulated more proline in M. rigidula and M. rotata under water stress than non-mycorrhizal plants. Conversely, mycorrhizal plants nodulated by the GM strain accumulated less proline in response to both AM colonization and drought. These results indicated changes in the synthesis of this nitrogenous osmoregulator product associated with microbial inoculation and drought tolerance. Mycorrhizal plants nodulated by the GM Sinorhizobium strain seem to suffer less from the detrimental effect of water stress, since under water limitation relative plant growth, percentage of AM colonization, root dry weight and the highest R/S ratio remained the same. The fact that GM nodulated plants are better adapted to drought stress could be of practical interest and the management of GM microorganism inoculation may be crucial for biotechnological approaches to improving crop yield in dry environments.

  18. Metabolic engineering of mannitol production in Lactococcus lactis: influence of overexpression of mannitol 1-phosphate dehydrogenase in different genetic backgrounds.

    PubMed

    Wisselink, H Wouter; Mars, Astrid E; van der Meer, Pieter; Eggink, Gerrit; Hugenholtz, Jeroen

    2004-07-01

    To obtain a mannitol-producing Lactococcus lactis strain, the mannitol 1-phosphate dehydrogenase gene (mtlD) from Lactobacillus plantarum was overexpressed in a wild-type strain, a lactate dehydrogenase(LDH)-deficient strain, and a strain with reduced phosphofructokinase activity. High-performance liquid chromatography and (13)C nuclear magnetic resonance analysis revealed that small amounts (<1%) of mannitol were formed by growing cells of mtlD-overexpressing LDH-deficient and phosphofructokinase-reduced strains, whereas resting cells of the LDH-deficient transformant converted 25% of glucose into mannitol. Moreover, the formed mannitol was not reutilized upon glucose depletion. Of the metabolic-engineering strategies investigated in this work, mtlD-overexpressing LDH-deficient L. lactis seemed to be the most promising strain for mannitol production.

  19. Continuous diffraction of molecules and disordered molecular crystals

    PubMed Central

    Yefanov, Oleksandr M.; Ayyer, Kartik; White, Thomas A.; Barty, Anton; Morgan, Andrew; Mariani, Valerio; Oberthuer, Dominik; Pande, Kanupriya

    2017-01-01

    The intensities of far-field diffraction patterns of orientationally aligned molecules obey Wilson statistics, whether those molecules are in isolation (giving rise to a continuous diffraction pattern) or arranged in a crystal (giving rise to Bragg peaks). Ensembles of molecules in several orientations, but uncorrelated in position, give rise to the incoherent sum of the diffraction from those objects, modifying the statistics in a similar way as crystal twinning modifies the distribution of Bragg intensities. This situation arises in the continuous diffraction of laser-aligned molecules or translationally disordered molecular crystals. This paper develops the analysis of the intensity statistics of such continuous diffraction to obtain parameters such as scaling, beam coherence and the number of contributing independent object orientations. When measured, continuous molecular diffraction is generally weak and accompanied by a background that far exceeds the strength of the signal. Instead of just relying upon the smallest measured intensities or their mean value to guide the subtraction of the background, it is shown how all measured values can be utilized to estimate the background, noise and signal, by employing a modified ‘noisy Wilson’ distribution that explicitly includes the background. Parameters relating to the background and signal quantities can be estimated from the moments of the measured intensities. The analysis method is demonstrated on previously published continuous diffraction data measured from crystals of photosystem II [Ayyer et al. (2016 ▸), Nature, 530, 202–206]. PMID:28808434

  20. Pre-Clinical Efficacy and Safety of Experimental Vaccines Based on Non-Replicating Vaccinia Vectors against Yellow Fever

    PubMed Central

    Schäfer, Birgit; Holzer, Georg W.; Joachimsthaler, Alexandra; Coulibaly, Sogue; Schwendinger, Michael; Crowe, Brian A.; Kreil, Thomas R.; Barrett, P. Noel; Falkner, Falko G.

    2011-01-01

    Background Currently existing yellow fever (YF) vaccines are based on the live attenuated yellow fever virus 17D strain (YFV-17D). Although, a good safety profile was historically attributed to the 17D vaccine, serious adverse events have been reported, making the development of a safer, more modern vaccine desirable. Methodology/Principal Findings A gene encoding the precursor of the membrane and envelope (prME) protein of the YFV-17D strain was inserted into the non-replicating modified vaccinia virus Ankara and into the D4R-defective vaccinia virus. Candidate vaccines based on the recombinant vaccinia viruses were assessed for immunogenicity and protection in a mouse model and compared to the commercial YFV-17D vaccine. The recombinant live vaccines induced γ-interferon-secreting CD4- and functionally active CD8-T cells, and conferred full protection against lethal challenge already after a single low immunization dose of 105 TCID50. Surprisingly, pre-existing immunity against wild-type vaccinia virus did not negatively influence protection. Unlike the classical 17D vaccine, the vaccinia virus-based vaccines did not cause mortality following intracerebral administration in mice, demonstrating better safety profiles. Conclusions/Significance The non-replicating recombinant YF candidate live vaccines induced a broad immune response after single dose administration, were effective even in the presence of a pre-existing immunity against vaccinia virus and demonstrated an excellent safety profile in mice. PMID:21931732

  1. Rheological characterization of geopolymer binder modified by organic resins

    NASA Astrophysics Data System (ADS)

    Cekalová, M.; Kovárík, T.; Rieger, D.

    2017-01-01

    The purpose of this study is going to investigate properties of alkali-activated powder (calcined kaoilinitic clay and granulated blast furnace slag) prepared as a geopolymer paste and modified by various amount of organic resin. Hybrid organic-inorganic binders were prepared as a mix of organic resin and geopolymer inorganic paste under vacuum conditions. The process of solidification was investigated by measurements of storage (G’) and loss modulus ( G’) in torsion. The measurement was conducted in oscillatory mode by constant strain of 0.01 %. This strain is set in linear visco-elastic region for minimization influence of paste structure. The effect of organic resin is presented and determined by changes of viscosity (‘n*), modules in torsion and tangent of loss angle (tan 8). Results indicate that addition of organic resin significantly affects the initial viscosity and hardening kinetics.

  2. Revitalizing genetically-modified mouse strains using frozen-thawed sperm after up to 192 h of refrigerated epididymis transportation.

    PubMed

    Moreno-Del Val, Gonzalo; Muñoz-Robledano, Patricia

    2017-10-01

    In the scientific interchange of genetically-modified mouse strains the transportation of refrigerated epididymis has several advantages over the transportation of live animals, especially with regard to the 3R (replacement, reduction and refinement) principles. The major limiting factor is the duration of the transportation. Previous reports have shown that sperm collected from transported epididymis maintained their fertility for around 72 h, but there are no published data with longer transportation times, and this window of time may be too short, especially for international shipments and where locations are not well connected. In this study live pups were born using frozen-thawed sperm after up to 192 h (8 days) of transportation, using a special in vitro fertilization design which resulted in a fertilization rate of 10.5%.

  3. Use of Attenuated but Metabolically Competent Salmonella as a Probiotic To Prevent or Treat Salmonella Infection

    PubMed Central

    Sabag-Daigle, Anice; Blunk, Henry M.; Gonzalez, Juan F.; Steidley, Brandi L.; Boyaka, Prosper N.

    2016-01-01

    Salmonella enterica is among the most burdensome of foodborne disease agents. There are over 2,600 serovars that cause a range of disease manifestations ranging from enterocolitis to typhoid fever. While there are two vaccines in use in humans to protect against typhoid fever, there are none that prevent enterocolitis. If vaccines preventing enterocolitis were to be developed, they would likely protect against only one or a few serovars. In this report, we tested the hypothesis that probiotic organisms could compete for the preferred nutrient sources of Salmonella and thus prevent or treat infection. To this end, we added the fra locus, which encodes a utilization pathway for the Salmonella-specific nutrient source fructose-asparagine (F-Asn), to the probiotic bacterium Escherichia coli Nissle 1917 (Nissle) to increase its ability to compete with Salmonella in mouse models. We also tested a metabolically competent, but avirulent, Salmonella enterica serovar Typhimurium mutant for its ability to compete with wild-type Salmonella. The modified Nissle strain became more virulent and less able to protect against Salmonella in some instances. On the other hand, the modified Salmonella strain was safe and effective in preventing infection with wild-type Salmonella. While we tested for efficacy only against Salmonella Typhimurium, the modified Salmonella strain may be able to compete metabolically with most, if not all, Salmonella serovars, representing a novel approach to control of this pathogen. PMID:27185789

  4. DNA repair properties of Escherichia coli tif-1, recAo281 and lexA1 strains deficient in single-strand DNA binding protein.

    PubMed

    Whittier, R F; Chase, J W

    1983-01-01

    Mutations affecting single-strand DNA binding protein (SSB) impair induction of mutagenic (SOS) repair. To further investigate the role of SSB in SOS induction and DNA repair, isogenic strains were constructed combining the ssb+, ssb-1 or ssb-113 alleles with one or more mutations known to alter regulation of damage inducible functions. As is true in ssb+ strains tif-1 (recA441) was found to allow thermal induction of prophage lambda + and Weigle reactivation in ssb-1 and ssb-113 strains. Furthermore, tif-1 decreased the UV sensitivity of the ssb-113 strain slightly and permitted UV induction of prophage lambda + at 30 degrees C. Strains carrying the recAo281 allele were also constructed. This mutation causes high constitutive levels of RecA protein synthesis and relieves much of the UV sensitivity conferred by lexA- alleles without restoring SOS (error-prone) repair. In contrast, the recAo281 allele failed to alleviate the UV sensitivity associated with either ssb- mutation. In a lexA1 recAo281 background the ssb-1 mutation increased the extent of postirradiation DNA degradation and concommitantly increased UV sensitivity 20-fold to the level exhibited by a recA1 strain. The ssb-113 mutation also increased UV sensitivity markedly in this background but did so without greatly increasing postirradiation DNA degradation. These results suggest a direct role for SSB in recombinational repair apart from and in addition to its role in facilitating induction of the recA-lexA regulon.

  5. Cyclomodulins in Urosepsis Strains of Escherichia coli▿

    PubMed Central

    Dubois, Damien; Delmas, Julien; Cady, Anne; Robin, Frédéric; Sivignon, Adeline; Oswald, Eric; Bonnet, Richard

    2010-01-01

    Determinants of urosepsis in Escherichia coli remain incompletely defined. Cyclomodulins (CMs) are a growing functional family of toxins that hijack the eukaryotic cell cycle. Four cyclomodulin types are actually known in E. coli: cytotoxic necrotizing factors (CNFs), cycle-inhibiting factor (Cif), cytolethal distending toxins (CDTs), and the pks-encoded toxin. In the present study, the distribution of CM-encoding genes and the functionality of these toxins were investigated in 197 E. coli strains isolated from patients with community-acquired urosepsis (n = 146) and from uninfected subjects (n = 51). This distribution was analyzed in relation to the phylogenetic background, clinical origin, and antibiotic resistance of the strains. It emerged from this study that strains harboring the pks island and the cnf1 gene (i) were strongly associated with the B2 phylogroup (P, <0.001), (ii) frequently harbored both toxin-encoded genes in phylogroup B2 (33%), and (iii) were predictive of a urosepsis origin (P, <0.001 to 0.005). However, the prevalences of the pks island among phylogroup B2 strains, in contrast to those of the cnf1 gene, were not significantly different between fecal and urosepsis groups, suggesting that the pks island is more important for the colonization process and the cnf1 gene for virulence. pks- or cnf1-harboring strains were significantly associated with susceptibility to antibiotics (amoxicillin, cotrimoxazole, and quinolones [P, <0.001 to 0.043]). Otherwise, only 6% and 1% of all strains harbored the cdtB and cif genes, respectively, with no particular distribution by phylogenetic background, antimicrobial susceptibility, or clinical origin. PMID:20375237

  6. Evaluation of industrial Saccharomyces cerevisiae strains as the chassis cell for second-generation bioethanol production

    PubMed Central

    Li, Hongxing; Wu, Meiling; Xu, Lili; Hou, Jin; Guo, Ting; Bao, Xiaoming; Shen, Yu

    2015-01-01

    To develop a suitable Saccharomyces cerevisiae industrial strain as a chassis cell for ethanol production using lignocellulosic materials, 32 wild-type strains were evaluated for their glucose fermenting ability, their tolerance to the stresses they might encounter in lignocellulosic hydrolysate fermentation and their genetic background for pentose metabolism. The strain BSIF, isolated from tropical fruit in Thailand, was selected out of the distinctly different strains studied for its promising characteristics. The maximal specific growth rate of BSIF was as high as 0.65 h−1 in yeast extract peptone dextrose medium, and the ethanol yield was 0.45 g g−1 consumed glucose. Furthermore, compared with other strains, this strain exhibited superior tolerance to high temperature, hyperosmotic stress and oxidative stress; better growth performance in lignocellulosic hydrolysate; and better xylose utilization capacity when an initial xylose metabolic pathway was introduced. All of these results indicate that this strain is an excellent chassis strain for lignocellulosic ethanol production. PMID:25616171

  7. Strain engineering of van der Waals heterostructures.

    PubMed

    Vermeulen, Paul A; Mulder, Jefta; Momand, Jamo; Kooi, Bart J

    2018-01-18

    Modifying the strain state of solids allows control over a plethora of functional properties. The weak interlayer bonding in van der Waals (vdWaals) materials such as graphene, hBN, MoS 2 , and Bi 2 Te 3 might seem to exclude strain engineering, since strain would immediately relax at the vdWaals interfaces. Here we present direct observations of the contrary by showing growth of vdWaals heterostructures with persistent in-plane strains up to 5% and we show that strain relaxation follows a not yet reported process distinctly different from strain relaxation in three-dimensionally bonded (3D) materials. For this, 2D bonded Bi 2 Te 3 -Sb 2 Te 3 and 2D/3D bonded Bi 2 Te 3 -GeTe multilayered films are grown using Pulsed Laser Deposition (PLD) and their structure is monitored in situ using Reflective High Energy Electron Diffraction (RHEED) and post situ analysis is performed using Transmission Electron Microscopy (TEM). Strain relaxation is modeled and found to solely depend on the layer being grown and its initial strain. This insight demonstrates that strain engineering of 2D bonded heterostructures obeys different rules than hold for epitaxial 3D materials and opens the door to precise tuning of the strain state of the individual layers to optimize functional performance of vdWaals heterostructures.

  8. Methodology optimization and diversification for the investigation of virulence potential in Haemophilus influenzae clinical strains.

    PubMed

    Giucă, Mihaela Cristina; Străuţ, Monica; Surdeanu, Maria; Nica, Maria; Ungureanu, Vasilica; Mihăescu, Grigore

    2011-01-01

    Ten Haemophilus influenzae strains were isolated from patients aged between 1.6 - 24 years, with various diagnoses (acute meningitis, acute upper respiratory infection, otitis media and acute sinusitis). Identification was based on phenotypic and molecular characteristics; antibiotic susceptibility testing was performed by diffusion method according to CLSI standards 2011 for seven antibiotics. The results of molecular testing showed that all the studied strains produced an amplicon of 1000 bp with ompP2 primers indicating that all strains were H. influenzae. For six strains, the PCR amplicon obtained with bexA specific primers, proving that the strains were capsulated. The results of phenotypic testing showed that four strains were ampicillin nonsusceptible and (beta-lactamase-positive. The virulence potential of H. influenzae clinical strains was investigated by phenotypic methods, including the assessment of the soluble virulence factors on specific media containing the biochemical substratum for the investigated enzymatic factor, as well as the adherence and invasion capacity to HeLa cells monolayer using Cravioto modified method. The studied strains exhibited mainly a diffuse adherence pattern and different adherence indexes. Interestingly, two strains isolated from the same pacient (blood and CSF) showed a different degree of invasiveness, the strain isolated from blood being 20 times more invasive than the one isolated from CSF.

  9. Implementation of Improved Transverse Shear Calculations and Higher Order Laminate Theory Into Strain Rate Dependent Analyses of Polymer Matrix Composites

    NASA Technical Reports Server (NTRS)

    Zhu, Lin-Fa; Kim, Soo; Chattopadhyay, Aditi; Goldberg, Robert K.

    2004-01-01

    A numerical procedure has been developed to investigate the nonlinear and strain rate dependent deformation response of polymer matrix composite laminated plates under high strain rate impact loadings. A recently developed strength of materials based micromechanics model, incorporating a set of nonlinear, strain rate dependent constitutive equations for the polymer matrix, is extended to account for the transverse shear effects during impact. Four different assumptions of transverse shear deformation are investigated in order to improve the developed strain rate dependent micromechanics model. The validities of these assumptions are investigated using numerical and theoretical approaches. A method to determine through the thickness strain and transverse Poisson's ratio of the composite is developed. The revised micromechanics model is then implemented into a higher order laminated plate theory which is modified to include the effects of inelastic strains. Parametric studies are conducted to investigate the mechanical response of composite plates under high strain rate loadings. Results show the transverse shear stresses cannot be neglected in the impact problem. A significant level of strain rate dependency and material nonlinearity is found in the deformation response of representative composite specimens.

  10. Enhancing adhesion of yeast brewery strains to chamotte carriers through aminosilane surface modification.

    PubMed

    Berlowska, Joanna; Kregiel, Dorota; Ambroziak, Wojciech

    2013-07-01

    The adhesion of cells to solid supports is described as surface-dependent, being largely determined by the properties of the surface. In this study, ceramic surfaces modified using different organosilanes were tested for proadhesive properties using industrial brewery yeast strains in different physiological states. Eight brewing strains were tested: bottom-fermenting Saccharomyces pastorianus and top-fermenting Saccharomyces cerevisiae. To determine adhesion efficiency light microscopy, scanning electron microscopy and the fluorymetric method were used. Modification of chamotte carriers by 3-(3-anino-2-hydroxy-1-propoxy) propyldimethoxysilane and 3-(N, N-dimethyl-N-2-hydroxyethyl) ammonium propyldimethoxysilane groups increased their biomass load significantly.

  11. Controlling nanoscale acoustic strains in silicon using chirped femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Tzianaki, E.; Bakarezos, M.; Tsibidis, G. D.; Petrakis, S.; Loukakos, P. A.; Kosmidis, C.; Tatarakis, M.; Papadogiannis, N. A.

    2016-06-01

    The influence of femtosecond laser pulse chirp on laser-generated longitudinal acoustic strains in Si (100) monocrystal substrates is studied. Degenerate femtosecond pump-probe transient reflectivity measurements are performed using a layered structure of thin Ti transducer film on an Si substrate. Experimental results show that acoustic strains, manifested as strong Brillouin oscillations, are more effectively induced when negatively chirped femtosecond laser pulses pump the transducer. These results are theoretically supported by a modified thermo-mechanical model based on the combination of a revised two-temperature model and elasticity theory that takes into account the instantaneous frequency of the chirped femtosecond laser pump pulses.

  12. Transmission dynamics of two dengue serotypes with vaccination scenarios.

    PubMed

    González Morales, N L; Núñez-López, M; Ramos-Castañeda, J; Velasco-Hernández, J X

    2017-05-01

    In this work we present a mathematical model that incorporates two Dengue serotypes. The model has been constructed to study both the epidemiological trends of the disease and conditions that allow coexistence in competing strains under vaccination. We consider two viral strains and temporary cross-immunity with one vector mosquito population. Results suggest that vaccination scenarios will not only reduce disease incidence but will also modify the transmission dynamics. Indeed, vaccination and cross immunity period are seen to decrease the frequency and magnitude of outbreaks but in a differentiated manner with specific effects depending upon the interaction vaccine and strain type. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Isolation of plasmid from the blue-green alga Spirulina platensis

    NASA Astrophysics Data System (ADS)

    Qin, Song; Tong, Shun; Zhang, Peijun; Tseng, C. K.

    1993-09-01

    CCC plasmid was isolated from an economically important blue-green alga — Spirulina platensis (1.7×106 dalton from the S6 strain and 1.2×106 dalton from the F3 strain) using a rapid method based on ultrasonic disruption of algal cells and alkaline removal of chromosomal DNA. The difference in the molecular weight of the CCC DNAs from the two strains differing in form suggests that plasmid may be related with the differentiation of algal form. This modified method, which does not use any lysozyme, is a quick and effective method of plasmid isolation, especially for filamentous blue-green algae.

  14. Analysis of genetic variation in Ganoderma Lucidum after space flight

    NASA Astrophysics Data System (ADS)

    Qi, Jian-Jun; Ma, Rong-Cai; Chen, Xiang-Dong; Lan, Jin

    A modified CTAB method was used in the extraction of total cellular DNA of Ganoderma lucidum. Four strains Cx, Ch, C3 and C4, and their counterparts, four space flown strains Sx, Xh, S3 and S4, were analysed by amplified fragment length polymorphism (AFLP) with several primer combinations. Polymorphic bands were detected between Sx and Cx, S3 and C3, respectively. Somatic incompatibility tests further confirmed their heterogeneity. However, no disparity between Sh and Ch, S4 and C4 was detectable. The results suggest that spaceflight may be used to accelerate breeding of Ganoderma lucidum strains for commercial cultivation.

  15. A description of the mechanical behavior of composite solid propellants based on molecular theory

    NASA Technical Reports Server (NTRS)

    Landel, R. F.

    1976-01-01

    Both the investigation and the representation of the stress-strain response (including rupture) of gum and filled elastomers can be based on a simple functional statement. Internally consistent experiments are used to sort out the effects of time, temperature, strain and crosslink density on gum rubbers. All effects are readily correlated and shown to be essentially independent of the elastomer when considered in terms of non-dimensionalized stress, strain and time. A semiquantitative molecular theory is developed to explain this result. The introduction of fillers modifies the response, but, guided by the framework thus provided, their effects can be readily accounted for.

  16. An experimental investigation of Iosipescu specimen for composite materials

    NASA Technical Reports Server (NTRS)

    Ho, H.; Tsai, M. Y.; Morton, J.; Farley, G. L.

    1991-01-01

    A detailed experimental evaluation of the Iosipescu specimen tested in the modified Wyoming fixture is presented. Moire interferometry is employed to determine the deformation of unidirectional and cross-ply graphite-epoxy specimens. The results of the moire experiments are compared to those from the traditional strain-gage method. It is shown that the strain-gage readings from one surface of a specimen together with corresponding data from moire interferometry on the opposite face documented an extreme sensitivity of some fiber orientations to twisting. A localized hybrid analysis is introduced to perform efficient reduction of moire data, producing whole-field strain distributions in the specimen test sections.

  17. Combining inhibitor tolerance and D-xylose fermentation in industrial Saccharomyces cerevisiae for efficient lignocellulose-based bioethanol production.

    PubMed

    Demeke, Mekonnen M; Dumortier, Françoise; Li, Yingying; Broeckx, Tom; Foulquié-Moreno, María R; Thevelein, Johan M

    2013-08-26

    In addition to efficient pentose utilization, high inhibitor tolerance is a key trait required in any organism used for economically viable industrial bioethanol production with lignocellulose biomass. Although recent work has succeeded in establishing efficient xylose fermentation in robust industrial Saccharomyces cerevisiae strains, the resulting strains still lacked sufficient inhibitor tolerance for efficient sugar fermentation in lignocellulose hydrolysates. The aim of the present work was to combine high xylose fermentation activity and high inhibitor tolerance in a single industrial yeast strain. We have screened 580 yeast strains for high inhibitor tolerance using undetoxified acid-pretreated spruce hydrolysate and identified a triploid industrial baker's yeast strain as having the highest inhibitor tolerance. From this strain, a mating competent diploid segregant with even higher inhibitor tolerance was obtained. It was crossed with the recently developed D-xylose fermenting diploid industrial strain GS1.11-26, with the Ethanol Red genetic background. Screening of 819 diploid segregants from the tetraploid hybrid resulted in two strains, GSF335 and GSF767, combining high inhibitor tolerance and efficient xylose fermentation. In a parallel approach, meiotic recombination of GS1.11-26 with a haploid segregant of Ethanol Red and screening of 104 segregants resulted in a similar inhibitor tolerant diploid strain, GSE16. The three superior strains exhibited significantly improved tolerance to inhibitors in spruce hydrolysate, higher glucose consumption rates, higher aerobic growth rates and higher maximal ethanol accumulation capacity in very-high gravity fermentation, compared to GS1.11-26. In complex medium, the D-xylose utilization rate by the three superior strains ranged from 0.36 to 0.67 g/g DW/h, which was lower than that of GS1.11-26 (1.10 g/g DW/h). On the other hand, in batch fermentation of undetoxified acid-pretreated spruce hydrolysate, the three superior strains showed comparable D-xylose utilization rates as GS1.11-26, probably because of their higher inhibitor tolerance. They produced up to 23% more ethanol compared to Ethanol Red. We have successfully constructed three superior industrial S. cerevisiae strains that combine efficient D-xylose utilization with high inhibitor tolerance. Since the background strain Ethanol Red has a proven record of successful industrial application, the three new superior strains have strong potential for direct application in industrial bioethanol production.

  18. Cyclic tensile response of a pre-tensioned polyurethane

    NASA Astrophysics Data System (ADS)

    Nie, Yizhou; Liao, Hangjie; Chen, Weinong W.

    2018-05-01

    In the research reported in this paper, we subject a polyurethane to uniaxial tensile loading at a quasi-static strain rate, a high strain rate and a jumping strain rate where the specimen is under quasi-static pre-tension and is further subjected to a dynamic cyclic loading using a modified Kolsky tension bar. The results obtained at the quasi-static and high strain rate clearly show that the mechanical response of this material is significantly rate sensitive. The rate-jumping experimental results show that the response of the material behavior is consistent before jumping. After jumping the stress-strain response of the material does not jump to the corresponding high-rate curve. Rather it approaches the high-rate curve asymptotically. A non-linear hyper-viscoelastic (NLHV) model, after having been calibrated by monotonic quasi-static and high-rate experimental results, was found to be capable of describing the material tensile behavior under such rate jumping conditions.

  19. Implementation of Laminate Theory Into Strain Rate Dependent Micromechanics Analysis of Polymer Matrix Composites

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.

    2000-01-01

    A research program is in progress to develop strain rate dependent deformation and failure models for the analysis of polymer matrix composites subject to impact loads. Previously, strain rate dependent inelastic constitutive equations developed to model the polymer matrix were implemented into a mechanics of materials based micromechanics method. In the current work, the computation of the effective inelastic strain in the micromechanics model was modified to fully incorporate the Poisson effect. The micromechanics equations were also combined with classical laminate theory to enable the analysis of symmetric multilayered laminates subject to in-plane loading. A quasi-incremental trapezoidal integration method was implemented to integrate the constitutive equations within the laminate theory. Verification studies were conducted using an AS4/PEEK composite using a variety of laminate configurations and strain rates. The predicted results compared well with experimentally obtained values.

  20. Effects of strain on the electronic structure, superconductivity, and nematicity in FeSe studied by angle-resolved photoemission spectroscopy

    NASA Astrophysics Data System (ADS)

    Phan, G. N.; Nakayama, K.; Sugawara, K.; Sato, T.; Urata, T.; Tanabe, Y.; Tanigaki, K.; Nabeshima, F.; Imai, Y.; Maeda, A.; Takahashi, T.

    2017-06-01

    One of central issues in iron-based superconductors is the role of structural change to the superconducting transition temperature (Tc). It was found in FeSe that the lattice strain leads to a drastic increase in Tc, accompanied by suppression of nematic order. By angle-resolved photoemission spectroscopy on tensile- or compressive-strained and strain-free FeSe, we experimentally show that the in-plane strain causes a marked change in the energy overlap (Δ Eh -e ) between the hole and electron pockets in the normal state. The change in Δ Eh -e modifies the Fermi-surface volume, leading to a change in Tc. Furthermore, the strength of nematicity is also found to be characterized by Δ Eh -e . These results suggest that the key to understanding the phase diagram is the fermiology and interactions linked to the semimetallic band overlap.

  1. Characterization of a turbomolecular-pumped magnetic sector mass spectrometer

    NASA Technical Reports Server (NTRS)

    Mehta, Narinder K.

    1988-01-01

    A Perkin Elmer MGA-1200, turbomolecular-pumped, magnetic sector, multiple gas analyzer mass spectrometer with modified inlet for fast response was characterized for the analysis of hydrogen, helium, oxygen and argon in nitrogen and helium background gases. This instrument was specially modified for the Vanderberg AFB SLC-6 Hydrogen Disposal Test Program, as a part of the Hydrogen Sampling System (H2S2). Linearity, precision, drift, detection limits and accuracy among other analytical parameters for each of the background gas were studied to evaluate the performance of the instrument. The result demonstrates that H2S2 mass spectrometer is a stable instrument and can be utilized for the quantitative analytical determination of hydrogen, helium, oxygen and argon in nitrogen and helium background gases.

  2. Overview of latex modified concrete overlays : final report.

    DOT National Transportation Integrated Search

    1984-01-01

    Twelve bridges with latex modified concrete (LMC) overlays ranging in age from new to 13 years were studied and their general condition found to be good. The half-cell and chloride data were inconclusive because background data were not available for...

  3. Job Strain and Cognitive Decline: A Prospective Study of the Framingham Offspring Cohort

    PubMed Central

    Agbenyikey, W; Karasek, R; Cifuentes, M; Wolf, PA; Seshadri, S; Taylor, JA; Beiser, AS; Au, R

    2017-01-01

    Background Workplace stress is known to be related with many behavioral and disease outcomes. However, little is known about its prospective relationship with measures of cognitive decline. Objective To investigate the association of job strain, psychological demands and job control on cognitive decline. Methods Participants from Framingham Offspring cohort (n=1429), were assessed on job strain, and received neuropsychological assessment approximately 15 years and 21 years afterwards. Results High job strain and low control were associated with decline in verbal learning and memory. Job strain was associated with decline in word recognition skills. Active job and passive job predicted decline in verbal learning and memory relative to low strain jobs in the younger subgroup. Active job and demands were positively associated with abstract reasoning skills. Conclusions Job strain and job control may infuence decline in cognitive performance. PMID:25890602

  4. Segregation of genes from donor strain during the production of recombinant congenic strains.

    PubMed

    van Zutphen, L F; Den Bieman, M; Lankhorst, A; Demant, P

    1991-07-01

    Recombinant congenic strains (RCS) constitute a set of inbred strains which are designed to dissect the genetic control of multigenic traits, such as tumour susceptibility or disease resistance. Each RCS contains a small fraction of the genome of a common donor strain, while the majority of genes stem from a common background strain. We tested at two stages of the inbreeding process in 20 RCS, derived from BALB/cHeA and STS/A, to see whether alleles from the STS/A donor strain are distributed over the RCS in a ratio as would theoretically be expected. Four marker genes (Pep-3; Pgm-1; Gpi-1 and Es-3) located at 4 different chromosomes were selected and the allelic distribution was tested after 3-4 and after 12 generations of inbreeding. The data obtained do not significantly deviate from the expected pattern, thus supporting the validity of the concept of RCS.

  5. Strain-Engineered Graphene Grown on Hexagonal Boron Nitride by Molecular Beam Epitaxy

    PubMed Central

    Summerfield, Alex; Davies, Andrew; Cheng, Tin S.; Korolkov, Vladimir V.; Cho, YongJin; Mellor, Christopher J.; Foxon, C. Thomas; Khlobystov, Andrei N.; Watanabe, Kenji; Taniguchi, Takashi; Eaves, Laurence; Novikov, Sergei V.; Beton, Peter H.

    2016-01-01

    Graphene grown by high temperature molecular beam epitaxy on hexagonal boron nitride (hBN) forms continuous domains with dimensions of order 20 μm, and exhibits moiré patterns with large periodicities, up to ~30 nm, indicating that the layers are highly strained. Topological defects in the moiré patterns are observed and attributed to the relaxation of graphene islands which nucleate at different sites and subsequently coalesce. In addition, cracks are formed leading to strain relaxation, highly anisotropic strain fields, and abrupt boundaries between regions with different moiré periods. These cracks can also be formed by modification of the layers with a local probe resulting in the contraction and physical displacement of graphene layers. The Raman spectra of regions with a large moiré period reveal split and shifted G and 2D peaks confirming the presence of strain. Our work demonstrates a new approach to the growth of epitaxial graphene and a means of generating and modifying strain in graphene. PMID:26928710

  6. Isolation and characterization of a novel native Bacillus thuringiensis strain BRC-HZM2 capable of degrading chlorpyrifos.

    PubMed

    Wu, Songqing; Peng, Yan; Huang, Zhangmin; Huang, Zhipeng; Xu, Lei; Ivan, Gelbič; Guan, Xiong; Zhang, Lingling; Zou, Shuangquan

    2015-03-01

    Studies were carried out to isolate chlorpyrifos degrading Bacillus thuringiensis (Bt) strains from chlorpyrifos-contaminated samples. Six Bt strains (isolation rate 2.7%) were isolated by modified sodium acetate antibiotic heat treatment, and one novel strain (BRC-HZM2) was selected for further analysis. Phenotype and phylogeny analysis of this strain was conducted on the basis of biochemical reactions, antibiotic sensitivity, 16s rRNA genes, plasmid profile, insecticidal crystal protein profiles, and PCR-RFLP for cry and cyt genes. The degradation rate of chlorpyrifos in liquid culture was estimated during 48 h of incubation for the isolate BRC-HZM2. More than 50% of the initial chlorpyrifos concentration degraded within 12 h, 88.9% after 48 h. These results highlight the potential of the Bt strain for biological control and the bioremediation of environments contaminated with chlorpyrifos. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Quasi-bound states in strained graphene

    NASA Astrophysics Data System (ADS)

    Bahamon, Dario; Qi, Zenan; Park, Harold; Pareira, Vitor; Campbell, David

    In this work, we explore the possibility of manipulating electronic states in graphene nanostructures by mechanical means. Specifically, we use molecular dynamics and tight-binding models to access the electronic and transport properties of strained graphene nanobubbles and graphene kirigami. We establish that low energy electrons can be confined in the arms of the kirigami and within the nanobubbles; under different load conditions the coupling between confined states and continuous states is modified creating different conductance line-shapes.

  8. Life without tRNAIle-lysidine synthetase: translation of the isoleucine codon AUA in Bacillus subtilis lacking the canonical tRNA2Ile

    PubMed Central

    Köhrer, Caroline; Mandal, Debabrata; Gaston, Kirk W.; Grosjean, Henri; Limbach, Patrick A.; RajBhandary, Uttam L.

    2014-01-01

    Translation of the isoleucine codon AUA in most prokaryotes requires a modified C (lysidine or agmatidine) at the wobble position of tRNA2Ile to base pair specifically with the A of the AUA codon but not with the G of AUG. Recently, a Bacillus subtilis strain was isolated in which the essential gene encoding tRNAIle-lysidine synthetase was deleted for the first time. In such a strain, C34 at the wobble position of tRNA2Ile is expected to remain unmodified and cells depend on a mutant suppressor tRNA derived from tRNA1Ile, in which G34 has been changed to U34. An important question, therefore, is how U34 base pairs with A without also base pairing with G. Here, we show (i) that unlike U34 at the wobble position of all B. subtilis tRNAs of known sequence, U34 in the mutant tRNA is not modified, and (ii) that the mutant tRNA binds strongly to the AUA codon on B. subtilis ribosomes but only weakly to AUG. These in vitro data explain why the suppressor strain displays only a low level of misreading AUG codons in vivo and, as shown here, grows at a rate comparable to that of the wild-type strain. PMID:24194599

  9. Chemical, Physical, and Mechanical Characterization of Isocyanate Cross-linked Amine-Modified Silica Aerogels

    NASA Technical Reports Server (NTRS)

    Katti, Atul; Shimpi, Nilesh; Roy, Samit; Lu, Hongbing; Fabrizio, Eve F.; Dass, Amala; Capadona, Lynn A.; Leventis, Nicholas

    2006-01-01

    We describe a new mechanically strong lightweight porous composite material obtained by encapsulating the skeletal framework of amine-modified silica aerogels with polyurea. The conformal polymer coating preserves the mesoporous structure of the underlying silica framework and the thermal conductivity remains low at 0.041 plus or minus 0.001 W m(sup -1 K(sup -1). The potential of the new cross-linked silica aerogels for load-carrying applications was determined through characterization of their mechanical behavior under compression, three-point bending, and dynamic mechanical analysis (DMA). A primary glass transition temperature of 130 C was identified through DMA. At room temperature, results indicate a hyperfoam behavior where in compression cross-linked aerogels are linearly elastic under small strains (less than 4%) and then exhibit yield behavior (until 40% strain), followed by densification and inelastic hardening. At room temperature the compressive Young's modulus and the Poisson's ratio were determined to be 129 plus or minus 8 MPa and 0.18, respectively, while the strain at ultimate failure is 77% and the average specific compressive stress at ultimate failure is 3.89 x 10(exp 5) N m kg(sup -1). The specific flexural strength is 2.16 x 10(exp 4) N m kg(sup -1). Effects on the compressive behavior of strain rate and low temperature were also evaluated.

  10. The hydrolytic activity of esterases in the yeast Saccharomyces cerevisiae is strain dependent.

    PubMed

    Kwolek-Mirek, Magdalena; Bednarska, Sabina; Zadrąg-Tęcza, Renata; Bartosz, Grzegorz

    2011-11-01

    Ester precursors of fluorogenic or chromogenic probes are often employed in studies of yeast cell biology. This study was aimed at a comparison of the ability of several commonly used laboratory wild-type Saccharomyces cerevisiae strains to hydrolyse the following model esters: fluorescein diacetate, 2-naphthyl acetate, PNPA (p-nitrophenyl acetate) and AMQI (7-acetoxy-1-methylquinolinum iodide). In all the strains, the esterase activity was localized mainly to the cytosol. Considerable differences in esterase activity were observed between various wild-type laboratory yeast strains. The phase of growth also contributed to the variation in esterase activity of the yeast. This diversity implies the need for caution in using intracellularly hydrolysed probes for a comparison of yeast strains with various genetic backgrounds.

  11. Watch out for your TRP1 marker: the effect of TRP1 gene on the growth at high and low temperatures in budding yeast.

    PubMed

    Leng, Gang; Song, Kiwon

    2016-05-01

    TRP1 is a frequently used auxotrophic marker for genetic modifications and selections in trp(-) budding yeast strains, including the commonly used wild-type strain W303a. However, we found that introduction of the TRP1 gene into a trp(-) strain significantly affected vegetative growth at low and high temperatures. Therefore, caution should be needed when working in a trp(-) background strain and using the TRP1 marker to study stress response phenotypes, particularly when analyzing temperature sensitivities. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Neuropeptide S alters anxiety, but not depression-like behaviour in Flinders Sensitive Line rats: a genetic animal model of depression.

    PubMed

    Wegener, Gregers; Finger, Beate C; Elfving, Betina; Keller, Kirsten; Liebenberg, Nico; Fischer, Christina W; Singewald, Nicolas; Slattery, David A; Neumann, Inga D; Mathé, Aleksander A

    2012-04-01

    Neuropeptide S (NPS) and its receptor (NPSR) have been implicated in the mediation of anxiolytic-like behaviour in rodents. However, little knowledge is available regarding the NPS system in depression-related behaviours, and whether NPS also exerts anxiolytic effects in an animal model of psychopathology. Therefore, the aim of this work was to characterize the effects of NPS on depression- and anxiety-related parameters, using male and female rats in a well-validated animal model of depression: the Flinders Sensitive Line (FSL), their controls, the Flinders Resistant Line (FRL), and Sprague-Dawley (SD) rats. We found that FSL showed greater immobility in the forced swim test (FST) than FRL, confirming their phenotype. However, NPS did not affect depression-related behaviour in any rat line. No significant differences in baseline anxiety levels between the FSL and FRL strains were observed, but FSL and FRL rats displayed less anxiety-like behaviour compared to SD rats. NPS decreased anxiety-like behaviour on the elevated plus-maze in all strains. The expression of the NPSR in the amygdala, periventricular hypothalamic nucleus, and hippocampus was equal in all male strains, although a trend towards reduced expression within the amygdala was observed in FSL rats compared to SD rats. In conclusion, NPS had a marked anxiolytic effect in FSL, FRL and SD rats, but did not modify the depression-related behaviour in any strain, in spite of the significant differences in innate level between the strains. These findings suggest that NPS specifically modifies anxiety behaviour but cannot overcome/reverse a genetically mediated depression phenotype.

  13. Zebrafish (Danio rerio) as a screen for attenuation of Lancefield group C streptococci and a model for streptococcal pathogenesis.

    PubMed

    Borst, L B; Patterson, S K; Lanka, S; Suyemoto, M M; Maddox, C W

    2013-05-01

    Group C streptococci are highly contagious pyogenic bacteria responsible for respiratory tract, lymph node, urogenital tract, and wound infections. Wild-type strains of Streptococcus equi ssp equi (S. equi) and Streptococcus equi ssp zooepidemicus (S. zoo) as well as a commercially available modified live vaccine strain of S. equi were evaluated for virulence in zebrafish. Survival times, histologic lesions, and relative gene expression were compared among groups. Based on the intramuscular route of infection, significantly shorter survival times were observed in fish infected with wild-type strain when compared to modified live vaccine and S. zoo strains. Histologically, S. zoo-infected fish demonstrated a marked increase in inflammatory infiltrates (predominantly macrophages) at the site of infection, as well as increased cellularity in the spleen and renal interstitium. In contrast, minimal cellular immune response was observed in S. equi-injected fish with local tissue necrosis and edema predominating. Based on whole comparative genomic hybridization, increased transcription of positive acute-phase proteins, coagulation factors, and antimicrobial peptides were observed in S. equi-injected fish relative to S. zoo-injected fish, while mediators of cellular inflammation, including CXC chemokines and granulin, were upregulated in S. zoo-injected fish relative to S. equi-injected fish. In a screen of 11 clinical isolates, S. equi strains with a single nucleotide deletion in the upstream region of szp, a known virulence factor of streptococci, were found to be significantly attenuated in zebrafish. These collective findings underscore the value of the zebrafish as a model of streptococcal pathogenesis.

  14. The influence of gender-specific loading patterns of the stop-jump task on anterior cruciate ligament strain.

    PubMed

    Weinhold, Paul S; Stewart, Jason-Dennis N; Liu, Hsin-Yi; Lin, Cheng-Feng; Garrett, William E; Yu, Bing

    2007-08-01

    Studies have shown that women are at higher risk of sustaining noncontact anterior cruciate ligament (ACL) injuries in specific sports. Recent gait studies of athletic tasks have documented that gender differences in knee movement, muscle activation, and external loading patterns exist. The objective of this study was to determine in a knee cadaver model if application of female-specific loading and movement patterns characterised in vivo for a stop-jump task cause higher ACL strains than male patterns. Gender-specific loading patterns of the landing phase of the vertical stop-jump task were applied to seven cadaver knees using published kinetic/kinematic results for recreational athletes. Loads applied consecutively included: tibial compression, quadriceps, hamstrings, external posterior tibial shear, and tibial torque. Knee flexion was fixed based on the kinematic data. Strain of the ACL was monitored by means of a differential variable reluctance transducer installed on the anterior-medial bundle of the ACL. The ACL strain was significantly increased (P<0.05) for the female loading pattern relative to the male loading pattern after the posterior tibial shear force was applied, and showed a similar trend (P=0.1) to be increased after the final tibial torque was applied. This study suggests that female motor control strategies used during the stop-jump task may place higher strains on the ACL than male strategies, thus putting females at greater risk of ACL injury. We believe these results suggest the potential effectiveness of using training programs to modify motor control strategies and thus modify the risk of injury.

  15. Advanced age-related denervation and fiber-type grouping in skeletal muscle of SOD1 knockout mice.

    PubMed

    Kostrominova, Tatiana Y

    2010-11-30

    In this study skeletal muscles from 1.5- and 10-month-old Cu/Zn superoxide dismutase (SOD1) homozygous knockout (JLSod1(-/-)) mice obtained from The Jackson Laboratory (C57Bl6/129SvEv background) were compared with muscles from age- and sex-matched heterozygous (JLSod1(+/-)) littermates. The results of this study were compared with previously published data on two different strains of Sod1(-/-) mice: one from Dr. Epstein's laboratory (ELSod1(-/-); C57Bl6 background) and the other from Cephalon, Inc. (CSod1(-/-); 129/CD-1 background). Grouping of succinate dehydrogenase-positive fibers characterized muscles of Sod1(-/-) mice from all three strains. The 10-month-old Sod1(-/-)C and JL mice displayed pronounced denervation of the gastrocnemius muscle, whereas the ELSod1(-/-) mice displayed a small degree of denervation at this age, but developed accelerated age-related denervation later on. Denervation markers were up-regulated in skeletal muscle of 10-month-old JLSod1(-/-) mice. This study is the first to show that metallothionein mRNA and protein expression was up-regulated in the skeletal muscle of 10-month-old JLSod1(-/-) mice and was mostly localized to the small atrophic muscle fibers. In conclusion, all three strains of Sod1(-/-) mice develop accelerated age-related muscle denervation, but the genetic background has significant influence on the progress of denervation. Copyright © 2010 Elsevier Inc. All rights reserved.

  16. Dynamic strain aging in the high-temperature low-cycle fatigue of SA508 Cl. 3 forging steel

    NASA Astrophysics Data System (ADS)

    Lee, Byung Ho; Kim, In Sup

    1995-10-01

    The effect of dynamic strain aging on cyclic stress response and fatigue resistance of ASME SA508 Cl.3 forging steel for nuclear reactor pressure vessels has been evaluated in the temperature range of room temperature to 500°C. Total strain ranges and strain rates were varied from 0.7 to 2.0% and from 4 × 10 -4 to 1 × 10 -2 s -1, respectively. The cyclic stress response depended on the testing temperature, strain rate, and range. Generally, the initial cyclic hardening was immediately followed by cyclic softening at all strain rates. However, at 300°C, the operating temperature of nuclear reactor pressure vessels, the variation of cyclic stress amplitude showed the primary and secondary hardening stages dependent on the strain rate and strain range. Dynamic strain aging was manifested by enhanced cyclic hardening, distinguished secondary hardening, and negative strain rate sensitivity. A modified cell shutting model was described for the onset of the secondary hardening due to the dynamic strain aging and it was in good agreement with the experimental results. Fatigue life increased in strain rate at all testing temperatures. Specifically the fatigue life was longer at the dynamic strain aging temperature. Further, the dynamic strain aging was easy to initiate the crack, while crack propagation was retarded by crack branching and suppression of plastic zone, hence the dynamic strain aging caused the improvement of fatigue resistance.

  17. Age, experience and genetic background influence treadmill walking in mice

    PubMed Central

    Wooley, Christine M.; Xing, Shuqin; Burgess, Robert W.; Cox, Gregory A.; Seburn, Kevin L.

    2009-01-01

    WOOLEY, C.M., S. XING, R.W. BURGESS, G.A. COX, AND K.L. SEBURN. Age, experience and genetic background influence treadmill walking in mice. PHYSIOL. BEHAV. XX(X), XXX-XXX, 2008 – The use of a treadmill to gather data for gait analysis in mice is a convenient, sensitive method to evaluate motor performance. However, evidence from several species, including mice, shows that treadmill locomotion is a novel task that is not equivalent to over ground locomotion and that may be particularly sensitive to the test environment and protocol. We investigated the effects of age, genetic background and repeated trials on treadmill walking in mice and show that these factors are important considerations in the interpretation of gait data. Specifically we report that as C57BL/6J (B6) mice age, the animals use progressively longer, less frequent strides to maintain the same walking speed. The increase is most rapid between 1 and 6 months of age and is explained, in part, by changes in size and weight. We also extended previous findings showing that repeat trials cause mice to modify their treadmill gait pattern. In general, B6 mice tend to take shorter, more frequent steps and adopt a wider dynamic stance with repeated walking trials. The nature and extent of the response changes with both the number and timing of the trials and was observed with inter-trial intervals as long as 3 months. Finally, we compared the gait pattern of an additional seven inbred strains of mice and found significant variation in the length and frequency of strides used to maintain the same walking speed. The combined results offer the bases for further mechanistic studies and can be used to guide optimal experimental design. PMID:19027767

  18. Analysis of Lactobacillus sakei Mutants Selected after Adaptation to the Gastrointestinal Tracts of Axenic Mice▿ †

    PubMed Central

    Chiaramonte, Fabrizio; Anglade, Patricia; Baraige, Fabienne; Gratadoux, Jean-Jacques; Langella, Philippe; Champomier-Vergès, Marie-Christine; Zagorec, Monique

    2010-01-01

    We recently showed that Lactobacillus sakei, a natural meat-borne lactic acid bacterium, can colonize the gastrointestinal tracts (GIT) of axenic mice but that this colonization in the intestinal environment selects L. sakei mutants showing modified colony morphology (small and rough) and cell shape, most probably resulting from the accumulation of various mutations that confer a selective advantage for persistence in the GIT. In the present study, we analyzed such clones, issued from three different L. sakei strains, in order to determine which functions were modified in the mutants. In the elongated filamentous cells of the rough clones, transmission electron microscopy (TEM) analysis showed a septation defect and dotted and slanted black bands, suggesting the presence of a helical structure around the cells. Comparison of the cytoplasmic and cell wall/membrane proteomes of the meat isolate L. sakei 23K and of one of its rough derivatives revealed a modified expression for 38 spots. The expression of six oxidoreductases, several stress proteins, and four ABC transporters was strongly reduced in the GIT-adapted strain, while the actin-like MreB protein responsible for cell shaping was upregulated. In addition, the expression of several enzymes involved in carbohydrate metabolism was modified, which may correlate with the observation of modified growth of mutants on various carbon sources. These results suggest that the modifications leading to a better adaptation to the GIT are pleiotropic and are characterized in a rough mutant by a different stress status, a cell wall modification, and modified use of energy sources, leading to an improved fitness for the colonization of the GIT. PMID:20208026

  19. Processing and Characterization of a Novel Distributed Strain Sensor Using Carbon Nanotube-Based Nonwoven Composites

    PubMed Central

    Dai, Hongbo; Thostenson, Erik T.; Schumacher, Thomas

    2015-01-01

    This paper describes the development of an innovative carbon nanotube-based non-woven composite sensor that can be tailored for strain sensing properties and potentially offers a reliable and cost-effective sensing option for structural health monitoring (SHM). This novel strain sensor is fabricated using a readily scalable process of coating Carbon nanotubes (CNT) onto a nonwoven carrier fabric to form an electrically-isotropic conductive network. Epoxy is then infused into the CNT-modified fabric to form a free-standing nanocomposite strain sensor. By measuring the changes in the electrical properties of the sensing composite the deformation can be measured in real-time. The sensors are repeatable and linear up to 0.4% strain. Highest elastic strain gage factors of 1.9 and 4.0 have been achieved in the longitudinal and transverse direction, respectively. Although the longitudinal gage factor of the newly formed nanocomposite sensor is close to some metallic foil strain gages, the proposed sensing methodology offers spatial coverage, manufacturing customizability, distributed sensing capability as well as transverse sensitivity. PMID:26197323

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

    Song, Bo; Nelson, Kevin; Jin, Helena

    Iridium alloys have been utilized as structural materials for certain high-temperature applications, due to their superior strength and ductility at elevated temperatures. The mechanical properties, including failure response at high strain rates and elevated temperatures of the iridium alloys need to be characterized to better understand high-speed impacts at elevated temperatures. A DOP-26 iridium alloy has been dynamically characterized in compression at elevated temperatures with high-temperature Kolsky compression bar techniques. However, the dynamic high-temperature compression tests were not able to provide sufficient dynamic high-temperature failure information of the iridium alloy. In this study, we modified current room-temperature Kolsky tension barmore » techniques for obtaining dynamic tensile stress-strain curves of the DOP-26 iridium alloy at two different strain rates (~1000 and ~3000 s-1) and temperatures (~750°C and ~1030°C). The effects of strain rate and temperature on the tensile stress-strain response of the iridium alloy were determined. The DOP-26 iridium alloy exhibited high ductility in stress-strain response that strongly depended on both strain rate and temperature.« less

  1. Influence of Austenite Stability on Steel Low Cycle Fatigue Response

    NASA Astrophysics Data System (ADS)

    Lehnhoff, G. R.; Findley, K. O.

    Austenitic steels were subjected to tensile and total strain controlled, fully reversed axial low cycle fatigue (LCF) testing to determine the influence of stacking fault energy on austenite stability, or resistance to strain induced martensitic transformation during tensile and fatigue deformation. Expected differences in stacking fault energy were achieved by modifying alloys with different amounts of silicon and aluminum. Al alloying was found to promote martensite formation during both tensile and LCF loading, while Si was found to stabilize austenite. Martensite formation increases tensile work hardening rates, though Si additions also increase the work hardening rate without martensite transformation. Similarly, secondary cyclic strain hardening during LCF is attributed to strain induced martensite formation, but Si alloying resulted in less secondary cyclic strain hardening. The amount of secondary cyclic hardening scales linearly with martensite fraction and depends only on the martensite fraction achieved and not on the martensite (i.e. parent austenite) chemistry. Martensite formation was detrimental to LCF lives at all strain amplitudes tested, although the total amount of martensitic transformation during LCF did not always monotonically increase with strain amplitude nor correlate to the amount of tensile transformation.

  2. FK506 biosynthesis is regulated by two positive regulatory elements in Streptomyces tsukubaensis

    PubMed Central

    2012-01-01

    Background FK506 (Tacrolimus) is an important immunosuppressant, produced by industrial biosynthetic processes using various Streptomyces species. Considering the complex structure of FK506, it is reasonable to expect complex regulatory networks controlling its biosynthesis. Regulatory elements, present in gene clusters can have a profound influence on the final yield of target product and can play an important role in development of industrial bioprocesses. Results Three putative regulatory elements, namely fkbR, belonging to the LysR-type family, fkbN, a large ATP-binding regulator of the LuxR family (LAL-type) and allN, a homologue of AsnC family regulatory proteins, were identified in the FK506 gene cluster from Streptomyces tsukubaensis NRRL 18488, a progenitor of industrial strains used for production of FK506. Inactivation of fkbN caused a complete disruption of FK506 biosynthesis, while inactivation of fkbR resulted in about 80% reduction of FK506 yield. No functional role in the regulation of the FK506 gene cluster has been observed for the allN gene. Using RT-PCR and a reporter system based on a chalcone synthase rppA, we demonstrated, that in the wild type as well as in fkbN- and fkbR-inactivated strains, fkbR is transcribed in all stages of cultivation, even before the onset of FK506 production, whereas fkbN expression is initiated approximately with the initiation of FK506 production. Surprisingly, inactivation of fkbN (or fkbR) does not abolish the transcription of the genes in the FK506 gene cluster in general, but may reduce expression of some of the tested biosynthetic genes. Finally, introduction of a second copy of the fkbR or fkbN genes under the control of the strong ermE* promoter into the wild type strain resulted in 30% and 55% of yield improvement, respectively. Conclusions Our results clearly demonstrate the positive regulatory role of fkbR and fkbN genes in FK506 biosynthesis in S. tsukubaensis NRRL 18488. We have shown that regulatory mechanisms can differ substantially from other, even apparently closely similar FK506-producing strains, reported in literature. Finally, we have demonstrated the potential of these genetically modified strains of S. tsukubaensis for improving the yield of fermentative processes for production of FK506. PMID:23083511

  3. A Technique for Facile and Precise Transfer of Mouse Embryos

    PubMed Central

    Sarvari, Ali; Naderi, Mohammad Mehdi; Sadeghi, Mohammad Reza; Akhondi, Mohammad Mehdi

    2013-01-01

    Background Successful Embryo Transfer (ET) technique is a fateful step of all efforts to achieve live births from in vitro produced embryos in assisted reproductive techniques or in knockout, transgenic or cloned animal projects. Small reproductive tract of mice and limitation of current techniques may not well satisfy the requirements for mass production of genetically modified mice. Genetic abnormalities of embryos, receptivity and uterine contractions, expulsion of embryos, blood, mucus or bacterial contamination on the transfer pipette tip, technical problems and even animal strain may affect embryo transfer outcome. Methods In this study, two techniques of embryo transfer in mice were compared. In conventional technique the oviduct wall was punctured with a 30-gauge needle and the loaded Pasteur pipette with embryos and medium was inserted into the hole. In new technique, embryos that were loaded in modified micropipette with minimal medium were transferred directly to the oviduct by manual piston micro-pump easily. Embryo viability was evaluated considering the percentage of live healthy newborns. Results Results of the two techniques were compared by t-test within the NPAR1WAY procedure of SAS software (ver. 9.2). The average live birth rates in the novel methods was significantly higher (42.4%) than the conventional method (21.7%, p<0.05). Conclusion In conclusion, using new embryo transfer technique improved birth rate by preventing embryos expulsion from the oviduct, saving time and easy transfer of embryos with minimum volume of medium. PMID:23626878

  4. A major locus on mouse chromosome 18 controls XX sex reversal in Odd Sex (Ods) mice.

    PubMed

    Qin, Yangjun; Poirier, Christophe; Truong, Cavatina; Schumacher, Armin; Agoulnik, Alexander I; Bishop, Colin E

    2003-03-01

    We have previously reported a dominant mouse mutant, Odd sex (Ods), in which XX Ods/+ mice on the FVB/N background show complete sex reversal, associated with expression of Sox9 in the fetal gonads. Remarkably, when crossed to the A/J strain approximately 95% of the (AXFVB) F(1) XX Ods/+ mice developed as fully fertile, phenotypic females, the remainder developing as males or hermaphrodites. Using a (AXFVB) F(2) population, we conducted a genome-wide linkage scan to identify the number and chromosomal location of potential Ods modifier genes. A single major locus termed Odsm1 was mapped to chromosome 18, tightly linked to D18Mit189 and D18Mit210. Segregation at this locus could account for the presence of sex reversal in 100% of XX Ods/+ mice which develop as males, for the absence of sex reversal in approximately 92% of XX Ods/+ mice which develop as females, and for the mixed sexual phenotype in approximately 72% of XX Ods/+ mice that develop with ambiguous genitalia. We propose that homozygosity for the FVB-derived allele strongly favors Ods sex reversal, whereas homozygosity for the A/J-derived allele inhibits it. In mice heterozygous at Odsm1, the phenotypic outcome, male, female or hermaphrodite, is determined by a complex interaction of several minor modifying loci. The close proximity of Smad2, Smad7 and Smad4 to D18Mit189/210 provides a potential mechanism through which Odsm1 might act.

  5. Phytocompounds and modulatory effects of Anacardium microcarpum (cajui) on antibiotic drugs used in clinical infections

    PubMed Central

    Barbosa-Filho, Valter M; Waczuk, Emily P; Leite, Nadghia F; Menezes, Irwin RA; da Costa, José GM; Lacerda, Sírleis R; Adedara, Isaac A; Coutinho, Henrique Douglas Melo; Posser, Thais; Kamdem, Jean P

    2015-01-01

    Background The challenge of antibiotic resistance and the emergence of new infections have generated considerable interest in the exploration of natural products from plant origins as combination therapy. In this context, crude ethanolic extract (CEE), ethyl acetate fraction (EAF), and methanolic fraction (MF) from Anacardium microcarpum were tested alone or in combination with antibiotics (amikacin, gentamicin, ciprofloxacin, and imipenem) against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. Methods Antibiotic resistance-modifying activity was performed using the microdilution method by determining the minimal inhibitory concentration (MIC). In addition, phytochemical prospecting analyses of tested samples were carried out. Results Our results indicated that all the extracts showed low antibacterial activity against multidrug-resistant strains (MIC =512 μg/mL). However, addition of CEE, EAF, and MF to the growth medium at the subinhibitory concentration (MIC/8=64 μg/mL) significantly modulated amikacin- and gentamicin-resistant E. coli 06. CEE and EAF also demonstrated a significant (P<0.001) synergism with imipenem against S. aureus. In contrast, MF antagonized the antibacterial effect of ciprofloxacin and gentamicin against P. aeruginosa 03 and S. aureus 10, respectively. Qualitative phytochemical analysis of the extracts revealed the presence of secondary metabolites including phenols, flavonoids, xanthones, chalcones, and tannin pyrogallates. Conclusion Taken together, our results suggest that A. microcarpum is a natural resource with resistance-modifying antibacterial activity that needs to be further investigated to overcome the present resistant-infection problem. PMID:26604695

  6. Effect of Serotype on Pneumococcal Competition in a Mouse Colonization Model.

    PubMed

    Trzciński, Krzysztof; Li, Yuan; Weinberger, Daniel M; Thompson, Claudette M; Cordy, Derrick; Bessolo, Andrew; Malley, Richard; Lipsitch, Marc

    2015-09-15

    Competitive interactions between Streptococcus pneumoniae strains during host colonization could influence the serotype distribution in nasopharyngeal carriage and pneumococcal disease. We evaluated the competitive fitness of strains of serotypes 6B, 14, 19A, 19F, 23F, and 35B in a mouse model of multiserotype carriage. Isogenic variants were constructed using clinical strains as the capsule gene donors. Animals were intranasally inoculated with a mixture of up to six pneumococcal strains of different serotypes, with separate experiments involving either clinical isolates or isogenic capsule-switch variants of clinical strain TIGR4. Upper-respiratory-tract samples were repeatedly collected from animals in order to monitor changes in the serotype ratios using quantitative PCR. A reproducible hierarchy of capsular types developed in the airways of mice inoculated with multiple strains. Serotype ranks in this hierarchy were similar among pneumococcal strains of different genetic backgrounds in different strains of mice and were not altered when tested under a range of host conditions. This rank correlated with the measure of the metabolic cost of capsule synthesis and in vitro measure of pneumococcal cell surface charge, both parameters considered to be predictors of serotype-specific fitness in carriage. This study demonstrates the presence of a robust competitive hierarchy of pneumococcal serotypes in vivo that is driven mainly, but not exclusively, by the capsule itself. Streptococcus pneumoniae (pneumococcus) is the leading cause of death due to respiratory bacterial infections but also a commensal frequently carried in upper airways. Available vaccines induce immune responses against polysaccharides coating pneumococcal cells, but with over 90 different capsular types (serotypes) identified, they can only target strains of the selected few serotypes most prevalent in disease. Vaccines not only protect vaccinated individuals against disease but also protect by reducing carriage of vaccine-targeted strains to induce herd effects across whole populations. Unfortunately, reduction in the circulation of vaccine-type strains is offset by increase in carriage and disease from nonvaccine strains, indicating the importance of competitive interactions between pneumococci in shaping the population structure of this pathogen. Here, we showed that the competitive ability of pneumococcal strains to colonize the host strongly depends on the type of capsular polysaccharide expressed by pneumococci and only to a lesser degree on strain or host genetic backgrounds or on variation in host immune responses. Copyright © 2015 Trzciński et al.

  7. Phenotypic evaluation and characterization of 21 industrial Saccharomyces cerevisiae yeast strains.

    PubMed

    Kong, In Iok; Turner, Timothy Lee; Kim, Heejin; Kim, Soo Rin; Jin, Yong-Su

    2018-02-01

    Microorganisms have been studied and used extensively to produce value-added fuels and chemicals. Yeasts, specifically Saccharomyces cerevisiae, receive industrial attention because of their well-known ability to ferment glucose and produce ethanol. Thousands of natural or genetically modified S. cerevisiae have been found in industrial environments for various purposes. These industrial strains are isolated from industrial fermentation sites, and they are considered as potential host strains for superior fermentation processes. In many cases, industrial yeast strains have higher thermotolerance, increased resistances towards fermentation inhibitors and increased glucose fermentation rates under anaerobic conditions when compared with laboratory yeast strains. Despite the advantages of industrial strains, they are often not well characterized. Through screening and phenotypic characterization of commercially available industrial yeast strains, industrial fermentation processes requiring specific environmental conditions may be able to select an ideal starting yeast strain to be further engineered. Here, we have characterized and compared 21 industrial S. cerevisiae strains under multiple conditions, including their tolerance to varying pH conditions, resistance to fermentation inhibitors, sporulation efficiency and ability to ferment lignocellulosic sugars. These data may be useful for the selection of a parental strain for specific biotechnological applications of engineered yeast. © FEMS 2018. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Electronic and phononic modulation of MoS2 under biaxial strain

    NASA Astrophysics Data System (ADS)

    Moghadasi, A.; Roknabadi, M. R.; Ghorbani, S. R.; Modarresi, M.

    2017-12-01

    Dichalcogenides of transition metals are attractive material due to its unique properties. In this work, it has been investigated the electronic band structure, phonon spectrum and heat capacity of MoS2 under the applied tensile and compressive biaxial strain using the density functional theory. The Molybdenum disulfide under compressive (tensile) strain up to 6% (10%) has stable atomic structure without any negative frequency in the phonon dispersion curves. The tensile biaxial strain reduces the energy gap in the electronic band structure and the optical-acoustic gap in phonon dispersion curves. The tensile biaxial strain also increases the specific heat capacity. On the other hand, the compressive biaxial strain in this material increases phonon gap and reduces the heat capacity and the electronic band gap. The phonon softening/hardening is reported for tensile/compressive biaxial strain in MoS2. We report phonon hardening for out of plane ZA mode in the presence of both tensile and compressive strains. Results show that the linear variation of specific heat with strain (CV ∝ε) and square dependency of specific heat with the temperature (CV ∝T2) for low temperature regime. The results demonstrate that the applied biaxial strain tunes the electronic energy gap and modifies the phonon spectrum of MoS2.

  9. Systems Approaches to Predict the Functions of Glycoside Hydrolases during the Life Cycle of Aspergillus niger Using Developmental Mutants ∆brlA and ∆flbA

    PubMed Central

    van Munster, Jolanda M.; Nitsche, Benjamin M.; Akeroyd, Michiel; Dijkhuizen, Lubbert; van der Maarel, Marc J. E. C.; Ram, Arthur F. J.

    2015-01-01

    Background The filamentous fungus Aspergillus niger encounters carbon starvation in nature as well as during industrial fermentations. In response, regulatory networks initiate and control autolysis and sporulation. Carbohydrate-active enzymes play an important role in these processes, for example by modifying cell walls during spore cell wall biogenesis or in cell wall degradation connected to autolysis. Results In this study, we used developmental mutants (ΔflbA and ΔbrlA) which are characterized by an aconidial phenotype when grown on a plate, but also in bioreactor-controlled submerged cultivations during carbon starvation. By comparing the transcriptomes, proteomes, enzyme activities and the fungal cell wall compositions of a wild type A. niger strain and these developmental mutants during carbon starvation, a global overview of the function of carbohydrate-active enzymes is provided. Seven genes encoding carbohydrate-active enzymes, including cfcA, were expressed during starvation in all strains; they may encode enzymes involved in cell wall recycling. Genes expressed in the wild-type during starvation, but not in the developmental mutants are likely involved in conidiogenesis. Eighteen of such genes were identified, including characterized sporulation-specific chitinases and An15g02350, member of the recently identified carbohydrate-active enzyme family AA11. Eight of the eighteen genes were also expressed, independent of FlbA or BrlA, in vegetative mycelium, indicating that they also have a role during vegetative growth. The ΔflbA strain had a reduced specific growth rate, an increased chitin content of the cell wall and specific expression of genes that are induced in response to cell wall stress, indicating that integrity of the cell wall of strain ΔflbA is reduced. Conclusion The combination of the developmental mutants ΔflbA and ΔbrlA resulted in the identification of enzymes involved in cell wall recycling and sporulation-specific cell wall modification, which contributes to understanding cell wall remodeling mechanisms during development. PMID:25629352

  10. Strains and stressors: an analysis of touchscreen learning in genetically diverse mouse strains.

    PubMed

    Graybeal, Carolyn; Bachu, Munisa; Mozhui, Khyobeni; Saksida, Lisa M; Bussey, Timothy J; Sagalyn, Erica; Williams, Robert W; Holmes, Andrew

    2014-01-01

    Touchscreen-based systems are growing in popularity as a tractable, translational approach for studying learning and cognition in rodents. However, while mouse strains are well known to differ in learning across various settings, performance variation between strains in touchscreen learning has not been well described. The selection of appropriate genetic strains and backgrounds is critical to the design of touchscreen-based studies and provides a basis for elucidating genetic factors moderating behavior. Here we provide a quantitative foundation for visual discrimination and reversal learning using touchscreen assays across a total of 35 genotypes. We found significant differences in operant performance and learning, including faster reversal learning in DBA/2J compared to C57BL/6J mice. We then assessed DBA/2J and C57BL/6J for differential sensitivity to an environmental insult by testing for alterations in reversal learning following exposure to repeated swim stress. Stress facilitated reversal learning (selectively during the late stage of reversal) in C57BL/6J, but did not affect learning in DBA/2J. To dissect genetic factors underlying these differences, we phenotyped a family of 27 BXD strains generated by crossing C57BL/6J and DBA/2J. There was marked variation in discrimination, reversal and extinction learning across the BXD strains, suggesting this task may be useful for identifying underlying genetic differences. Moreover, different measures of touchscreen learning were only modestly correlated in the BXD strains, indicating that these processes are comparatively independent at both genetic and phenotypic levels. Finally, we examined the behavioral structure of learning via principal component analysis of the current data, plus an archival dataset, totaling 765 mice. This revealed 5 independent factors suggestive of "reversal learning," "motivation-related late reversal learning," "discrimination learning," "speed to respond," and "motivation during discrimination." Together, these findings provide a valuable reference to inform the choice of strains and genetic backgrounds in future studies using touchscreen-based tasks.

  11. Funding innovation for treatment for rare diseases: adopting a cost-based yardstick approach

    PubMed Central

    2013-01-01

    Background Manufacturers justify the high prices for orphan drugs on the basis that the associated R&D costs must be spread over few patients. The proliferation of these drugs in the last three decades, combined with high prices commonly in excess of $100,000 per patient per year are placing a substantial strain on the budgets of drug plans in many countries. Do insurers spend a growing portion of their budgets on small patient populations, or leave vulnerable patients without coverage for valuable treatments? We suggest that a third option is present in the form of a cost-based regulatory mechanism. Methods This article explores the use of a cost-based price control mechanism for orphan drugs, adapted from the standard models applied in utilities regulation. Results and conclusions A rate-of-return style model, employing yardsticked cost allocations and a modified two-stage rate of return calculation could be effective in setting a new standard for orphan drugs pricing. This type of cost-based pricing would limit the costs faced by insurers while continuing to provide an efficient incentive for new drug development. PMID:24237605

  12. Endogenous retroviral insertion in Cryge in the mouse No3 cataract mutant

    PubMed Central

    Nag, Nabanita; Peterson, Katherine; Wyatt, Keith; Hess, Sonja; Ray, Sugata; Favor, Jack; Bogani, Debora; Lyon, Mary; Wistow, Graeme

    2007-01-01

    No3 (nuclear opacity 3) is a novel congenital nuclear cataract in mice. Microsatellite mapping placed the No3 locus on chromosome 1 between D1Mit480 (32cM) and D1Mit7 (41cM), a region containing seven crystallin genes; Cryba2 and the Cryga-Crygf cluster. Although polymorphic variants were observed, no candidate mutations were found for six of the genes. However, DNA walking identified a murine endogenous retrovirus (IAPLTR1: ERVK) insertion in exon 3 of Cryge, disrupting the coding sequence for γE-crystallin. Recombinant protein for the mutant γE was completely insoluble. The No3 cataract is mild compared with the effects of similar mutations of γE. Quantitative RT-PCR showed that γE/F mRNA levels are reduced in No3, suggesting that the relatively mild phenotype results from suppression of γE levels due to ERVK insertion. However, the severity of cataract is also strain dependent suggesting that genetic background modifiers also play a role in the development of opacity. PMID:17223009

  13. Improvements In Ethanologenic Escherichia Coli and Klebsiella Oxytoca

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

    Dr. David Nunn

    2010-09-30

    The current Verenium cellulosic ethanol process is based on the dilute-acid pretreatment of a biomass feedstock, followed by a two-stage fermentation of the pentose sugar-containing hydrolysate by a genetically modified ethanologenic Escherichia coli strain and a separate simultaneous saccharification-fermentation (SSF) of the cellulosic fraction by a genetically modified ethanologenic Klebsiella oxytoca strain and a fungal enzyme cocktail. In order to reduce unit operations and produce a fermentation beer with higher ethanol concentrations to reduce distillation costs, we have proposed to develop a simultaneous saccharification co-fermentation (SScF) process, where the fermentation of the pentose-containing hydrolysate and cellulosic fraction occurs within themore » same fermentation vessel. In order to accomplish this goal, improvements in the ethanologens must be made to address a number of issues that arise, including improved hydrolysate tolerance, co-fermentation of the pentose and hexose sugars and increased ethanol tolerance. Using a variety of approaches, including transcriptomics, strain adaptation, metagenomics and directed evolution, this work describes the efforts of a team of scientists from Verenium, University of Florida, Massachusetts Institute of Technology and Genomatica to improve the E. coli and K. oxytoca ethanologens to meet these requirements.« less

  14. Antibiotic-modifying activity of riachin, a non-cyanogenic cyanoglycoside extracted from Bauhinia pentandra.

    PubMed

    de Farias, Pablo Antonio Maia; Figueredo, Fernando Gomes; Lucas, Aline Maria Brito; de Moura, Rafael Barbosa; Coutinho, Henrique Douglas Melo; da Silva, Tania Maria Sarmento; Martin, Ana Luiza de Aguiar Rocha; Fonteles, Marta Maria de França

    2015-01-01

    The search for new active compounds from the Brazilian flora has intensified in recent years, especially for new drugs with antibiotic potential. Accordingly, the aim of this study was to determine whether riachin has antibiotic activity in itself or is able to modulate the activity of conventional antibiotics. A non-cyanogenic cyanoglycoside known as riachin was isolated from Bauhinia pentandra, and was tested alone and in combination with three antibiotics (clindamycin, amikacin, and gentamicin) against multiresistant bacterial strains (Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus). Riachin did not show significant antibiotic activity when tested alone against any strain (P>0.05). However, when combined with conventional antibiotics, it showed drug-modifying activity against strains of S. aureus exposed to clindamycin (P<0.001) as well as against P. aeruginosa exposed to amikacin (P<0.001). Although riachin did not show direct antibiotic activity, it had synergistic activity when combined with amikacin or clindamycin. The mechanism of action of this synergism is under investigation. The results of this work demonstrate that some substances of natural origin can enhance the effectiveness of certain antibiotics, which means a substantial reduction in the drug dose required and possibly in consequent adverse events for patients.

  15. Chimeric GII.4 norovirus virus-like-particle-based vaccines induce broadly blocking immune responses.

    PubMed

    Debbink, Kari; Lindesmith, Lisa C; Donaldson, Eric F; Swanstrom, Jesica; Baric, Ralph S

    2014-07-01

    There is currently no licensed vaccine for noroviruses, and development is hindered, in part, by an incomplete understanding of the host adaptive immune response to these highly heterogeneous viruses and rapid GII.4 norovirus molecular evolution. Emergence of a new predominant GII.4 norovirus strain occurs every 2 to 4 years. To address the problem of GII.4 antigenic variation, we tested the hypothesis that chimeric virus-like particle (VLP)-based vaccine platforms, which incorporate antigenic determinants from multiple strains into a single genetic background, will elicit a broader immune response against contemporary and emergent strains. Here, we compare the immune response generated by chimeric VLPs to that of parental strains and a multivalent VLP cocktail. Results demonstrate that chimeric VLPs induce a more broadly cross-blocking immune response than single parental VLPs and a similar response to a multivalent GII.4 VLP cocktail. Furthermore, we show that incorporating epitope site A alone from one strain into the background of another is sufficient to induce a blockade response against the strain donating epitope site A. This suggests a mechanism by which population-wide surveillance of mutations in a single epitope could be used to evaluate antigenic changes in order to identify potential emergent strains and quickly reformulate vaccines against future epidemic strains as they emerge in human populations. Noroviruses are gastrointestinal pathogens that infect an estimated 21 million people per year in the United States alone. GII.4 noroviruses account for >70% of all outbreaks, making them the most clinically important genotype. GII.4 noroviruses undergo a pattern of epochal evolution, resulting in the emergence of new strains with altered antigenicity over time, complicating vaccine design. This work is relevant to norovirus vaccine design as it demonstrates the potential for development of a chimeric VLP-based vaccine platform that may broaden the protective response against multiple GII.4 strains and proposes a potential reformulation strategy to control newly emergent strains in the human population. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  16. Genomic background-related activation of microglia and reduced β-amyloidosis in a mouse model of Alzheimer's disease.

    PubMed

    Fröhlich, Christina; Paarmann, Kristin; Steffen, Johannes; Stenzel, Jan; Krohn, Markus; Heinze, Hans-Jochen; Pahnke, Jens

    2013-03-01

    Alzheimer's disease (AD) is by far the most common neurodegenerative disease. AD is histologically characterized not only by extracellular senile plaques and vascular deposits consisting of β-amyloid (Aβ) but also by accompanying neuroinflammatory processes involving the brain's microglia. The importance of the microglia is still in controversial discussion, which currently favors a protective function in disease progression. Recent findings by different research groups highlighted the importance of strain-specific and mitochondria-specific genomic variations in mouse models of cerebral β-amyloidosis. Here, we want to summarize our previously presented data and add new results that draw attention towards the consideration of strain-specific genomic alterations in the setting of APP transgenes. We present data from APP-transgenic mice in commonly used C57Bl/6J and FVB/N genomic backgrounds and show a direct influence on the kinetics of Aβ deposition and the activity of resident microglia. Plaque size, plaque deposition rate and the total amount of Aβ are highest in C57Bl/6J mice as compared to the FVB/N genomic background, which can be explained at least partially by a reduced microglia activity towards amyloid deposits in the C57BL/6J strain.

  17. Listeriolysin S: A bacteriocin from epidemic Listeria monocytogenes strains that targets the gut microbiota.

    PubMed

    Quereda, Juan J; Meza-Torres, Jazmín; Cossart, Pascale; Pizarro-Cerdá, Javier

    2017-07-04

    Listeria monocytogenes is a Gram-positive food-borne pathogen that in humans may traverse the intestinal, placental and blood/brain barriers, causing gastroenteritis, abortions and meningitis. Crossing of these barriers is dependent on the bacterial ability to enter host cells, and several L. monocytogenes surface and secreted virulence factors are known to facilitate entry and the intracellular lifecycle. The study of L. monocytogenes strains associated to human listeriosis epidemics has revealed the presence of novel virulence factors. One such factor is Listeriolysin S, a thiazole/oxazole modified microcin that displays bactericidal activity and modifies the host microbiota during infection. Our recent results therefore highlight the interaction of L. monocytogenes with gut microbes as a crucial step in epidemic listeriosis. In this article, we will discuss novel implications for this family of toxins in the pathogenesis of diverse medically relevant microorganisms.

  18. Enhancing saccharification of wheat straw by mixing enzymes from genetically-modified Trichoderma reesei and Aspergillus niger.

    PubMed

    Jiang, Yanping; Duarte, Alexandra Vivas; van den Brink, Joost; Wiebenga, Ad; Zou, Gen; Wang, Chengshu; de Vries, Ronald P; Zhou, Zhihua; Benoit, Isabelle

    2016-01-01

    To increase the efficiency of enzymatic hydrolysis for plant biomass conversion into renewable biofuel and chemicals. By overexpressing the point mutation A824 V transcriptional activator Xyr1 in Trichoderma reesei, carboxymethyl cellulase, cellobiosidase and β-D-glucosidase activities of the best mutant were increased from 1.8 IU/ml, 0.1 IU/ml and 0.05 IU/ml to 4.8 IU/ml, 0.4 IU/ml and 0.3 IU/ml, respectively. The sugar yield of wheat straw saccharification by combining enzymes from this mutant and the Aspergillus niger genetically modified strain ΔcreA/xlnR c/araR c was improved up to 7.5 mg/ml, a 229 % increase compared to the combination of wild type strains. Mixing enzymes from T. reesei and A. niger combined with the genetic modification of transcription factors is a promising strategy to increase saccharification efficiency.

  19. Fatty acid production in genetically modified cyanobacteria

    PubMed Central

    Liu, Xinyao; Sheng, Jie; Curtiss III, Roy

    2011-01-01

    To avoid costly biomass recovery in photosynthetic microbial biofuel production, we genetically modified cyanobacteria to produce and secrete fatty acids. Starting with introducing an acyl–acyl carrier protein thioesterase gene, we made six successive generations of genetic modifications of cyanobacterium Synechocystis sp. PCC6803 wild type (SD100). The fatty acid secretion yield was increased to 197 ± 14 mg/L of culture in one improved strain at a cell density of 1.0 × 109 cells/mL by adding codon-optimized thioesterase genes and weakening polar cell wall layers. Although these strains exhibited damaged cell membranes at low cell densities, they grew more rapidly at high cell densities in late exponential and stationary phase and exhibited less cell damage than cells in wild-type cultures. Our results suggest that fatty acid secreting cyanobacteria are a promising technology for renewable biofuel production. PMID:21482809

  20. One library to make them all: streamlining the creation of yeast libraries via a SWAp-Tag strategy.

    PubMed

    Yofe, Ido; Weill, Uri; Meurer, Matthias; Chuartzman, Silvia; Zalckvar, Einat; Goldman, Omer; Ben-Dor, Shifra; Schütze, Conny; Wiedemann, Nils; Knop, Michael; Khmelinskii, Anton; Schuldiner, Maya

    2016-04-01

    The yeast Saccharomyces cerevisiae is ideal for systematic studies relying on collections of modified strains (libraries). Despite the significance of yeast libraries and the immense variety of available tags and regulatory elements, only a few such libraries exist, as their construction is extremely expensive and laborious. To overcome these limitations, we developed a SWAp-Tag (SWAT) method that enables one parental library to be modified easily and efficiently to give rise to an endless variety of libraries of choice. To showcase the versatility of the SWAT approach, we constructed and investigated a library of ∼1,800 strains carrying SWAT-GFP modules at the amino termini of endomembrane proteins and then used it to create two new libraries (mCherry and seamless GFP). Our work demonstrates how the SWAT method allows fast and effortless creation of yeast libraries, opening the door to new ways of systematically studying cell biology.

  1. Listeriolysin S: A bacteriocin from epidemic Listeria monocytogenes strains that targets the gut microbiota

    PubMed Central

    Quereda, Juan J.; Meza-Torres, Jazmín; Cossart, Pascale; Pizarro-Cerdá, Javier

    2017-01-01

    ABSTRACT Listeria monocytogenes is a Gram-positive food-borne pathogen that in humans may traverse the intestinal, placental and blood/brain barriers, causing gastroenteritis, abortions and meningitis. Crossing of these barriers is dependent on the bacterial ability to enter host cells, and several L. monocytogenes surface and secreted virulence factors are known to facilitate entry and the intracellular lifecycle. The study of L. monocytogenes strains associated to human listeriosis epidemics has revealed the presence of novel virulence factors. One such factor is Listeriolysin S, a thiazole/oxazole modified microcin that displays bactericidal activity and modifies the host microbiota during infection. Our recent results therefore highlight the interaction of L. monocytogenes with gut microbes as a crucial step in epidemic listeriosis. In this article, we will discuss novel implications for this family of toxins in the pathogenesis of diverse medically relevant microorganisms. PMID:28156183

  2. Blast Loading of Epoxy Panels Using a Shock Tube

    NASA Technical Reports Server (NTRS)

    Pankow, Mark; Waas, Anthony M.; Bednarcyk, Brett

    2010-01-01

    The high strain rate mechanical response of thin polymer plates has been studied using a modified shock tube. Diagnostics include the pressure-time history of the incident and reflected pulses and the use of digital image correlation (DIC) techniques to extract the time-history of the out-of-plane displacement distribution. Additionally, finite element models have been developed to understand the plate response and to validate and modify plate material constitutive models that have been proposed.

  3. Microorganisms having enhanced tolerance to inhibitors and stress

    DOEpatents

    Brown, Steven D.; Yang, Shihui

    2014-07-29

    The present invention provides genetically modified strains of microorganisms that display enhanced tolerance to stress and/or inhibitors such as sodium acetate and vanillin. The enhanced tolerance can be achieved by increasing the expression of a protein of the Sm-like superfamily such as a bacterial Hfq protein and a fungal Sm or Lsm protein. Further, the present invention provides methods of producing alcohol from biomass materials by using the genetically modified microorganisms of the present invention.

  4. In-phase and out-of-phase axial-torsional fatigue behavior of Haynes 188 at 760 C

    NASA Technical Reports Server (NTRS)

    Kalluri, Sreeramesh; Bonacuse, Peter J.

    1991-01-01

    Isothermal, in-phase and out-of-phase axial-torsional fatigue experiments have been conducted at 760 C on uniform gage section, thin-walled tubular specimens of a wrought cobalt-base superalloy, Haynes 188. Test-control and data acquisition were accomplished with a minicomputer. Fatigue lives of the in- and out-of-phase axial-torsional fatigue tests have been estimated with four different multiaxial fatigue life prediction models that were developed primarly for predicting axial-torsional fatigue lives at room temperature. The models investigated were: (1) the von Mises equivalent strain range; (2) the Modified Multiaxiality Factor Approach; (3) the Modified Smith-Watson-Topper Parameter; and (4) the critical shear plane method of Fatemi, Socie, and Kurath. In general, life predictions by the von Mises equivalent strain range model were within a factor of 2 for a majority of the tests and the predictions by the Modified Multiaxiality Factor Approach were within a factor of 2, while predictions of the Modified Smith-Watson-Topper Parameter and of the critical shear plane method of Fatemi, Socie, and Kurath were unconservative and conservative, respectively, by up to factors of 4. In some of the specimens tested under combined axial-torsional loading conditions, fatigue cracks initiated near extensometer indentations. Two design modifications have been proposed to the thin-walled tubular specimen to overcome this problem.

  5. Job strain and health-related quality of life in a national sample.

    PubMed

    Lerner, D J; Levine, S; Malspeis, S; D'Agostino, R B

    1994-10-01

    Studies of the health effects of job strain have focused on morbidity and mortality as outcomes. This is the first study to examine the relationship of job strain to more comprehensive health status measures that encompass health-related quality of life. In a national cross-sectional survey, 1319 working men and women, aged 18 through 64 years, completed a modified version of the Job Content Questionnaire that classified workers' jobs into four categories: high strain, passive, low strain, and active. Subjects also completed the Medical Outcomes Study Short-Form Health Survey and a health distress scale. Logistic regression analyses were employed that controlled for age, race/ethnicity, gender, and education. Both work and nonwork variables were included. Job strain was significantly associated with five of nine components of health-related quality of life: physical functioning, role functioning related to physical health, vitality, social functioning, and mental health. Job strain made a modest yet statistically significant contribution beyond the effects of chronic illness and psychosocial variables. The results provide justification for further investigating the role of job strain as an independent risk factor for health-related quality of life.

  6. Computational strategy for the solution of large strain nonlinear problems using the Wilkins explicit finite-difference approach

    NASA Technical Reports Server (NTRS)

    Hofmann, R.

    1980-01-01

    The STEALTH code system, which solves large strain, nonlinear continuum mechanics problems, was rigorously structured in both overall design and programming standards. The design is based on the theoretical elements of analysis while the programming standards attempt to establish a parallelism between physical theory, programming structure, and documentation. These features have made it easy to maintain, modify, and transport the codes. It has also guaranteed users a high level of quality control and quality assurance.

  7. Damage Accumulation in Advanced Metal Matrix Composites Under Thermal Cycling/Creep Loadings

    DTIC Science & Technology

    1993-09-01

    34transformation strain" (Eshelby, 1957) or * eigenstrain " (Mura, 1987). The stress-strain relation of a ccmposite predicted by the Tanaka-Mori model is given by a0o...a short fiber MMC can be calculated if the following eigenstrains are given in the domains of fiber (0l) and of the punched region encompassing the...and Mortenson modified the above model to account for partial relaxation by dislocation punching along the fiber axis. The eigenstrains based on this

  8. Recombination of mitochondrial DNA without selection pressure among compatible strains of the Aspergillus niger species aggregate.

    PubMed

    Tóth, B; Hamari, Z; Ferenczy, L; Varga, J; Kevei, F

    1998-03-01

    Previous mitochondrial transmission experiments between oligomycin-resistant and oligomycin-sensitive incompatible strains of the A. niger aggregate bearing various mtDNA RFLP profiles resulted in a great variety of mitochondrial recombinants under selection pressure. Apart from the recombinant mtDNAs, resistant clones harbouring unchanged RFLP profiles of resistant donor mtDNAs with the recipient nuclear backgrounds were rarely isolated. These strains were anastomosed with nuclearly isogenic oligomycin-sensitive recipient partners and the mitochondria of the resulting progeny were examined under non-selective conditions. These experiments provide insights into events which are possibly similar to those occurring in nature. The heterokaryons obtained formed both oligomycin-resistant and -sensitive sectors, most of which were found to be homoplasmons. Progenies harbouring oligomycin-resistant and -sensitive mtDNAs may originate either from individual recombination events or be due to parental segregation. MtDNA recombination might take place in the heterokaryons without selection by oligomycin. The most frequent recombinant types of mtDNA RFLP profiles were indistinguishable from those recombinant mtDNAs which were frequently obtained under selection pressure from directed transfer experiments between incompatible strains. We present evidence that mixed mitochondrial populations may influence the compatibility reactions in the presence of an isogenic nuclear background, that recombination may take place without selection pressure, and that the process does not require specific nuclear sequences of both parental strains.

  9. Drosophila melanogaster White Mutant w1118 Undergo Retinal Degeneration

    PubMed Central

    Ferreiro, María José; Pérez, Coralia; Marchesano, Mariana; Ruiz, Santiago; Caputi, Angel; Aguilera, Pedro; Barrio, Rosa; Cantera, Rafael

    2018-01-01

    Key scientific discoveries have resulted from genetic studies of Drosophila melanogaster, using a multitude of transgenic fly strains, the majority of which are constructed in a genetic background containing mutations in the white gene. Here we report that white mutant flies from w1118 strain undergo retinal degeneration. We observed also that w1118 mutants have progressive loss of climbing ability, shortened life span, as well as impaired resistance to various forms of stress. Retinal degeneration was abolished by transgenic expression of mini-white+ in the white null background w1118. We conclude that beyond the classical eye-color phenotype, mutations in Drosophila white gene could impair several biological functions affecting parameters like mobility, life span and stress tolerance. Consequently, we suggest caution and attentiveness during the interpretation of old experiments employing white mutant flies and when planning new ones, especially within the research field of neurodegeneration and neuroprotection. We also encourage that the use of w1118 strain as a wild-type control should be avoided. PMID:29354028

  10. Drosophila melanogaster White Mutant w 1118 Undergo Retinal Degeneration.

    PubMed

    Ferreiro, María José; Pérez, Coralia; Marchesano, Mariana; Ruiz, Santiago; Caputi, Angel; Aguilera, Pedro; Barrio, Rosa; Cantera, Rafael

    2017-01-01

    Key scientific discoveries have resulted from genetic studies of Drosophila melanogaster , using a multitude of transgenic fly strains, the majority of which are constructed in a genetic background containing mutations in the white gene. Here we report that white mutant flies from w 1118 strain undergo retinal degeneration. We observed also that w 1118 mutants have progressive loss of climbing ability, shortened life span, as well as impaired resistance to various forms of stress. Retinal degeneration was abolished by transgenic expression of mini-white + in the white null background w 1118 . We conclude that beyond the classical eye-color phenotype, mutations in Drosophila white gene could impair several biological functions affecting parameters like mobility, life span and stress tolerance. Consequently, we suggest caution and attentiveness during the interpretation of old experiments employing white mutant flies and when planning new ones, especially within the research field of neurodegeneration and neuroprotection. We also encourage that the use of w 1118 strain as a wild-type control should be avoided.

  11. Mob/oriT, a mobilizable site-specific recombination system for unmarked genetic manipulation in Bacillus thuringiensis and Bacillus cereus.

    PubMed

    Wang, Pengxia; Zhu, Yiguang; Zhang, Yuyang; Zhang, Chunyi; Xu, Jianyi; Deng, Yun; Peng, Donghai; Ruan, Lifang; Sun, Ming

    2016-06-10

    Bacillus thuringiensis and Bacillus cereus are two important species in B. cereus group. The intensive study of these strains at the molecular level and construction of genetically modified bacteria requires the development of efficient genetic tools. To insert genes into or delete genes from bacterial chromosomes, marker-less manipulation methods were employed. We present a novel genetic manipulation method for B. thuringiensis and B. cereus strains that does not leave selection markers. Our approach takes advantage of the relaxase Mob02281 encoded by plasmid pBMB0228 from Bacillus thuringiensis. In addition to its mobilization function, this Mob protein can mediate recombination between oriT sites. The Mob02281 mobilization module was associated with a spectinomycin-resistance gene to form a Mob-Spc cassette, which was flanked by the core 24-bp oriT sequences from pBMB0228. A strain in which the wild-type chromosome was replaced with the modified copy containing the Mob-Spc cassette at the target locus was obtained via homologous recombination. Thus, the spectinomycin-resistance gene can be used to screen for Mob-Spc cassette integration mutants. Recombination between the two oriT sequences mediated by Mob02281, encoded by the Mob-Spc cassette, resulted in the excision of the Mob-Spc cassette, producing the desired chromosomal alteration without introducing unwanted selection markers. We used this system to generate an in-frame deletion of a target gene in B. thuringiensis as well as a gene located in an operon of B. cereus. Moreover, we demonstrated that this system can be used to introduce a single gene or an expression cassette of interest in B. thuringiensis. The Mob/oriT recombination system provides an efficient method for unmarked genetic manipulation and for constructing genetically modified bacteria of B. thuringiensis and B. cereus. Our method extends the available genetic tools for B. thuringiensis and B. cereus strains.

  12. Virulence of Melissococcus plutonius and secondary invaders associated with European foulbrood disease of the honey bee.

    PubMed

    Lewkowski, Oleg; Erler, Silvio

    2018-05-24

    European foulbrood is a globally distributed brood disease affecting honey bees. It may lead to lethal infections of larvae and, in severe cases, even to colony collapse. Lately, a profound genetic and phenotypic diversity was documented for the causative agent Melissococcus plutonius. However, experimental work on the impact of diverse M. plutonius strains on hosts with different genetic background is completely lacking and the role of secondary invaders is poorly understood. Here, we address these issues and elucidate the impact and interaction of both host and pathogen on one another. Moreover, we try to unravel the role of secondary bacterial invasions in foulbrood-diseased larvae. We employed in vitro infections with honey bee larvae from queens with different genetic background and three different M. plutonius strains. Larvae infection experiments showed host-dependent survival dynamics although M. plutonius strain 49.3 consistently had the highest virulence. This pattern was also reflected in significantly reduced weights of 49.3 strain-infected larvae compared to the other treatments. No difference was found in groups additionally inoculated with a secondary invader (Enterococcus faecalis or Paenibacillus alvei) neither in terms of larval survival nor weight. These results suggest that host background contributes markedly to the course of the disease but virulence is mainly dependent on pathogen genotype. Secondary invaders following a M. plutonius infection do not increase disease lethality and therefore may just be a colonization of weakened and immunodeficient, or dead larvae. © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  13. Weight gain in mice on a high caloric diet and chronically treated with omeprazole depends on sex and genetic background.

    PubMed

    Saqui-Salces, Milena; Tsao, Amy C; Gillilland, Merritt G; Merchant, Juanita L

    2017-01-01

    The impact of omeprazole (OM), a widely used over-the-counter proton pump inhibitor, on weight gain has not been extensively explored. We examined what factors, e.g., diet composition, microbiota, genetic strain, and sex, might affect weight gain in mice fed a high caloric diet while on OM. Inbred C57BL/6J strain, a 50:50 hybrid (B6SJLF1/J) strain, and mice on a highly mixed genetic background were fed four diets: standard chow (STD, 6% fat), STD with 200 ppm OM (STD + O), a high-energy chow (HiE, 11% fat), and HiE chow with OM (HiE + O) for 17 wk. Metabolic analysis, body composition, and fecal microbiota composition were analyzed in C57BL/6J mice. Oral glucose tolerance tests were performed using mice on the mixed background. After 8 wk, female and male C57BL/6J mice on the HiE diets ate less, whereas males on the HiE diets compared with the STD diets gained weight. All diet treatments reduced energy expenditure in females but in males only those on the HiE + O diet. Gut microbiota composition differed in the C57BL/6J females but not the males. Hybrid B6SJLF1/J mice showed similar weight gain on all test diets. In contrast, mixed strain male mice fed a HiE + O diet gained ∼40% more weight than females on the same diet. In addition to increased weight gain, mixed genetic mice on the HiE + O diet cleared glucose normally but secreted more insulin. We concluded that sex and genetic background define weight gain and metabolic responses of mice on high caloric diets and OM. Copyright © 2017 the American Physiological Society.

  14. Gpd1 and Gpd2 Fine-Tuning for Sustainable Reduction of Glycerol Formation in Saccharomyces cerevisiae▿

    PubMed Central

    Hubmann, Georg; Guillouet, Stephane; Nevoigt, Elke

    2011-01-01

    Gpd1 and Gpd2 are the two isoforms of glycerol 3-phosphate dehydrogenase (GPDH), which is the rate-controlling enzyme of glycerol formation in Saccharomyces cerevisiae. The two isoenzymes play crucial roles in osmoregulation and redox balancing. Past approaches to increase ethanol yield at the cost of reduced glycerol yield have most often been based on deletion of either one or two isogenes (GPD1 and GPD2). While single deletions of GPD1 or GPD2 reduced glycerol formation only slightly, the gpd1Δ gpd2Δ double deletion strain produced zero glycerol but showed an osmosensitive phenotype and abolished anaerobic growth. Our current approach has sought to generate “intermediate” phenotypes by reducing both isoenzyme activities without abolishing them. To this end, the GPD1 promoter was replaced in a gpd2Δ background by two lower-strength TEF1 promoter mutants. In the same manner, the activity of the GPD2 promoter was reduced in a gpd1Δ background. The resulting strains were crossed to obtain different combinations of residual GPD1 and GPD2 expression levels. Among our engineered strains we identified four candidates showing improved ethanol yields compared to the wild type. In contrast to a gpd1Δ gpd2Δ double-deletion strain, these strains were able to completely ferment the sugars under quasi-anaerobic conditions in both minimal medium and during simultaneous saccharification and fermentation (SSF) of liquefied wheat mash (wheat liquefact). This result implies that our strains can tolerate the ethanol concentration at the end of the wheat liquefact SSF (up to 90 g liter−1). Moreover, a few of these strains showed no significant reduction in osmotic stress tolerance compared to the wild type. PMID:21724879

  15. Multigenome analysis identifies a worldwide distributed epidemic Legionella pneumophila clone that emerged within a highly diverse species

    PubMed Central

    Cazalet, Christel; Jarraud, Sophie; Ghavi-Helm, Yad; Kunst, Frank; Glaser, Philippe; Etienne; Buchrieser, Carmen

    2008-01-01

    Genomics can provide the basis for understanding the evolution of emerging, lethal human pathogens such as Legionella pneumophila, the causative agent of Legionnaires’ disease. This bacterium replicates within amoebae and persists in the environment as a free-living microbe. Among the many Legionella species described, L. pneumophila is associated with 90% of human disease and within the 15 serogroups (Sg), L. pneumophila Sg1 causes over 84% of Legionnaires’ disease worldwide. Why L. pneumophila Sg1 is so predominant is unknown. Here, we report the first comprehensive screen of the gene content of 217 L. pneumophila and 32 non-L. pneumophila strains isolated from humans and the environment using a Legionella DNA-array. Strikingly, we uncovered a high conservation of virulence- and eukaryotic-like genes, indicating strong environmental selection pressures for their preservation. No specific hybridization profile differentiated clinical and environmental strains or strains of different serogroups. Surprisingly, the gene cluster coding the determinants of the core and the O side-chain synthesis of the lipopolysaccaride (LPS cluster) determining Sg1 was present in diverse genomic backgrounds, strongly implicating the LPS of Sg1 itself as a principal cause of the high prevalence of Sg1 strains in human disease and suggesting that the LPS cluster can be transferred horizontally. Genomic analysis also revealed that L. pneumophila is a genetically diverse species, in part due to horizontal gene transfer of mobile genetic elements among L. pneumophila strains, but also between different Legionella species. However, the genomic background also plays a role in disease causation as demonstrated by the identification of a globally distributed epidemic strain exhibiting the genotype of the sequenced L. pneumophila strain Paris. PMID:18256241

  16. Complete genome sequence of Pelosinus sp. strain UFO1 assembled using single-molecule real-time DNA sequencing technology

    DOE PAGES

    Brown, Steven D.; Utturkar, Sagar M.; Magnuson, Timothy S.; ...

    2014-09-04

    Pelosinus fermentans strain R7 was isolated from Russian kaolin clays as the type strain and it can reduce Fe(III) during fermentative growth (1). Draft genome sequences for P. fermentans R7 and four strains from Hanford, Washington, USA, have been published (2–4). The P. fermentans 16S rRNA sequence dominated the lactate-based enrichment cultures from three geochemically contrasting soils from the Melton Branch Watershed, Oak Ridge, Tennessee, USA (5) and also at another stimulated, uraniumcontaminated field site near Oak Ridge (6). For the current work, strain UFO1 was isolated from pristine sediments at a background field site in Oak Ridge and characterizedmore » as facilitating U(VI) reduction and precipitation with phosphate (7).« less

  17. Developing the multiple stress-strain creep recovery (MS-SCR) test

    NASA Astrophysics Data System (ADS)

    Elnasri, Mahmoud; Airey, Gordon; Thom, Nick

    2018-04-01

    While most published work from Europe has been concerned with evaluating binders' resistance to rutting based on their stiffness (deformation resistance), work originating in the US has mainly been concerned with ranking binders based on their recoverability in a multiple stress form. This paper details the design of a new modified multiple stress-strain creep recovery (MS-SCR) test. The test is designed to evaluate binders' rutting resistance based on two rutting resistance mechanisms: stiffness and recoverability. A preliminary investigation is presented in this paper followed by details of the design of the new modified test. A 40/60 penetration grade bitumen and bitumen-filler mastics prepared with three filler concentrations (35%, 50%, and 65% filler content by mass of mastic) were tested. In addition, two polymer modified bitumens (PMBs) using the same base bitumen type were examined for validation. Two parameters are introduced to characterise the short and long recovery in the new test. In terms of stiffness, the test allows the behaviour of binders at different stress levels and loading cycles to be studied and produces a new parameter that can quantify the degree of modification. Finally, a relationship between nonlinearity and normal force in the test was investigated.

  18. Effect of thermal modification on rheological properties of polyethylene blends

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

    Siriprumpoonthum, Monchai; Nobukawa, Shogo; Yamaguchi, Masayuki, E-mail: m-yama@jaist.ac.jp

    2014-03-15

    We examined the effects of thermal modification under flow field on the rheological properties of linear low-density polyethylene (LLDPE) with high molecular weight, low-density polyethylene (LDPE), and their blends, without thermal stabilizer. Although structural changes during processing are not detected by size extrusion chromatography or nuclear magnetic resonance spectroscopy, linear viscoelastic properties changed greatly, especially for the LLDPE. A cross-linking reaction took place, leading to, presumably, star-shaped long-chain branches. Consequently, the modified LLDPE, having high zero-shear viscosity, became a thermorheologically complex melt. Moreover, it should be noted that the drawdown force, defined as the uniaxial elongational force at a constantmore » draw ratio, was significantly enhanced for the blends. Enhancement of elongational viscosity was also detected. The drawdown force and elongational viscosity are marked for the thermally modified blend as compared with those for the blend of thermally modified pure components. Intermolecular cross-linking reactions between LDPE and LLDPE, yielding polymers with more than two branch points per chain, result in marked strain-hardening in the elongational viscosity behavior even at small strain. The recovery curve of the oscillatory modulus after the shear modification is further evidence of a branched structure.« less

  19. Impact of novel shift handle laparoscopic tool on wrist ergonomics and task performance

    PubMed Central

    Yu, Denny; Lowndes, Bethany; Morrow, Missy; Kaufman, Kenton; Bingener, Juliane; Hallbeck, Susan

    2015-01-01

    Background Laparoscopic tool handles causing wrist flexion and extension more than 15° from neutral are considered “at-risk” for musculoskeletal strain. Therefore this study measured the impact of laparoscopic tool handle angles on wrist postures and task performance. Methods Eight surgeons performed standard and modified Fundamentals of Laparoscopic Surgery (FLS) tasks with laparoscopic tools. Tool A had three adjustable handle angle configurations, i.e., in-line 0° (A0), 30° (A30), and pistol-grip 70° (A70). Tool B was a fixed pistol-grip grasper. Participants performed FLS peg transfer, inverted peg transfer, and inverted circle-cut with each tool and handle angle. Inverted tasks were adapted from standard FLS tasks to simulate advanced tasks observed during abdominal wall surgeries, e.g., ventral hernia. Motion tracking, video-analysis, and modified NASA-TLX workload questionnaires were used to measure postures, performance (e.g., completion time and errors), and workload. Results Task performance did not differ among tools. For FLS peg transfer, self-reported physical workload was lower for B than A70, and mean wrist postures showed significantly higher flexion for in-line than pistol-grip tools (B and A70). For inverted peg transfer, workload was higher for all configurations. However, less time was spent in at-risk wrist postures for in-line (47%) than pistol-grip (93-94%), and most participants preferred Tool A. For inverted circle cut, workload did not vary across configurations, mean wrist posture was 10° closer to neutral for A0 than B, and median time in at-risk wrist postures was significantly less for A0 (43%) than B (87%). Conclusion The best ergonomic wrist positions for FLS (floor) tasks are provided by pistol-grip tools and for tasks on the abdominal wall (ventral surface) by in-line handles. Adjustable handle angle laparoscopic tools can reduce ergonomic risks for musculoskeletal strain and allow versatility for tasks alternating between the floor and ceiling positions in a surgical trainer without impacting performance. PMID:26541720

  20. Background noise measurements from jet exit vanes designed to reduced flow pulsations in an open-jet wind tunnel

    NASA Technical Reports Server (NTRS)

    Hoad, D. R.; Martin, R. M.

    1985-01-01

    Many open jet wind tunnels experience pulsations of the flow which are typically characterized by periodic low frequency velocity and pressure variations. One method of reducing these fluctuations is to install vanes around the perimeter of the jet exit to protrude into the flow. Although these vanes were shown to be effective in reducing the fluctuation content, they can also increase the test section background noise level. The results of an experimental acoustic program in the Langley 4- by 7-Meter Tunnel is presented which evaluates the effect on tunnel background noise of such modifications to the jet exit nozzle. Noise levels for the baseline tunnel configuration are compared with those for three jet exit nozzle modifications, including an enhanced noise reduction configuration that minimizes the effect of the vanes on the background noise. Although the noise levels for this modified vane configuration were comparable to baseline tunnel background noise levels in this facility, installation of these modified vanes in an acoustic tunnel may be of concern because the noise levels for the vanes could be well above background noise levels in a quiet facility.

Top