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microbiological contamination of digested products from anaerobic co-digestion of bovine manure and agricultural by-products.this study was performed to investigate the microbiological contamination of digestate product (dp) obtained from the anaerobic co-digestion of bovine manure and agricultural by-products.201121899580
Characteristics of the Shiga-toxin-producing enteroaggregative Escherichia coli O104:H4 German outbreak strain and of STEC strains isolated in Spain.A Shiga-toxin-producing Escherichia coli (STEC) strain belonging to serotype O104:H4, phylogenetic group B1 and sequence type ST678, with virulence features common to the enteroaggregative E. coli (EAEC) pathotype, was reported as the cause of the recent 2011 outbreak in Germany. The outbreak strain was determined to carry several virulence factors of extraintestinal pathogenic E. coli (ExPEC) and to be resistant to a wide range of antibiotics. There are only a few reports of serotype O104:H4, w ...201122101411
thermal denaturation produced degenerative proteins and interfered with ms for proteins dissolved in lysis buffer in proteomic analysis.in 1-de, proteins were traditionally mixed with the standard laemmli buffer and boiled for several minutes. recently, proteins dissolved in lysis buffer were used to produce better-resolved 2-de gels, but thermal denaturation procedure still remained in some proteomic analysis. to determine the detailed effects of thermal denaturation on sds-page and ms, both 1-de and 2-de were performed using proteins heated at 100°c for different periods of time, and 17 protein bands/spots were positively iden ...201121298662
udder pathogens and their resistance to antimicrobial agents in dairy cows in estonia.the goal of this study was to estimate the distribution of udder pathogens and their antibiotic resistance in estonia during the years 2007-2009.201121299911
variation of inflammatory dynamics and mediators in primiparous cows after intramammary challenge with escherichia coli.abstract: the objective of the current study was to investigate (i) the outcome of experimentally induced escherichia coli mastitis in primiparous cows during early lactation in relation with production of eicosanoids and inflammatory indicators, and (ii) the validity of thermography to evaluate temperature changes on udder skin surface after experimentally induced e. coli mastitis. nine primiparous holstein friesian cows were inoculated 24 ± 6 days (d) after parturition in both left quarters wi ...201121314974
the effect of lemongrass oil and its major components on clinical isolate mastitis pathogens and their mechanisms of action on staphylococcus aureus dmst 4745.the aims of this study were to investigate the antibacterial activity of lemongrass oil (lg) and its major components which were citral, geraniol and myrcene, against four strains of clinically isolated bovine mastitis pathogens, including staphylococcus aureus, streptococcus agalactiae, bacillus cereus and escherichia coli by the broth microdilution method, as well as their activity on s. aureus biofilm formation. attempts to clarify their mechanisms of action by investigation of the effects on ...201121316719
effects of cytoplasmic and periplasmic chaperones on secretory production of single-chain fv antibody in escherichia coli.the effects of cytoplasmic and periplasmic chaperones on the secretory production of an anti-bovine ribonuclease a single-chain variable fragment (scfv) 3a21 in escherichia coli were investigated. co-expression of a cytoplasmic chaperone, groel/es, dnak/dnaj/grpe, trigger factor, or secb with 3a21 scfv affected the proportions of antigen-binding activity in the cytoplasmic soluble fraction, the periplasmic fraction, and the extracellular medium, but there was no significant difference in the tot ...201121324738
effects of simulated dry and wet chilling and aging of beef fat and lean tissues on the reduction of escherichia coli o157:h7 and salmonella.the efficacy of dry and wet chilling and aging of beef as methods for the reduction of escherichia coli o157:h7 and salmonella on lean and fat tissues was studied. samples were obtained from a harvest facility prior to antimicrobial interventions and were inoculated with a cocktail mixture of e. coli o157:h7 or salmonella to achieve a target inoculation of 6 log cfu/cm(2). wet chilled and aged samples were then suspended, sprayed (10 °c) continuously for 15 min and then sprayed for 1 min every 1 ...201121333151
molecular characterization of antimicrobial resistance in gram-negative bacteria isolated from bovine mastitis in egypt.the aim of this study was to characterize the genetic basis of multidrug resistance in gram-negative bacteria isolated from bovine mastitis cases in egypt. multidrug resistance phenotypes were found in 34 of 112 (30.4%) gram-negative bacterial isolates, which harbored at least one antimicrobial resistance gene. the most prevalent multidrug-resistant (mdr) species were enterobacter cloacae (8 isolates, 7.1%), klebsiella pneumoniae (7 isolates, 6.3%), klebsiella oxytoca (7 isolates, 6.3%), escheri ...201121338385
differential gene expression of the toll-like receptor-4 cascade and neutrophil function in early- and mid-lactating dairy cows.during early lactation, neutrophils display several reduced immune functions. particularly, a delayed recruitment of neutrophils into the infected udder seems to be one of the underlying events involved in the severity of postpartum escherichia coli intramammary infections. the purpose of this study was to analyze the effect of in vitro chemotaxis and diapedesis on the expression of toll-like receptor-4 (tlr4)-related genes in bovine blood neutrophils isolated from 10 early-lactating (el) and 10 ...201121338793
in depth analysis of genes and pathways of the mammary gland involved in the pathogenesis of bovine escherichia coli-mastitis.bovine mastitis is one of the most costly and prevalent diseases affecting dairy cows worldwide. in order to develop new strategies to prevent escherichia coli-induced mastitis, a detailed understanding of the molecular mechanisms underlying the host immune response to an e. coli infection is necessary. to this end, we performed a global gene-expression analysis of mammary gland tissue collected from dairy cows that had been exposed to a controlled e. coli infection. biopsy samples of healthy an ...201121352611
efficacy of enterotoxigenic escherichia coli vaccine for bovine clinical mastitis.an enterotoxigenic escherichia coli (etec) vaccine designed to prevent diarrhoea was inoculated into dairy cows, and the occurrence of clinical mastitis was investigated for 2 years. half of 480 cows in five farms were subcutaneously inoculated with etec vaccine (imocolibov) twice with a 1-month interval in 2007 and 2008. fisher's exact test and survival (time to event) analysis with the log-rank test were used to compare vaccinates and controls. in 2007, there was no significant difference in t ...201121371362
in vitro effects of very low levels of aflatoxin b1 on free radicals production and bactericidal activity of bovine blood neutrophils.as one of the most potent and hazardous feed/food-originated mycotoxins, aflatoxin (af) b1 is regarded as a potent immunosuppressor in dairy cows. neutrophils (pmn), as key effector cells against pathogens, have a high potential to kill engulfed microbes. to investigate the in vitro effects of very low doses of afb1 on blood pmn functions, we examined the effects of biologically relevant concentrations of afb1 on the phagocytosis and non-phagocytosis dependent luminol, representative of mainly i ...201121377741
antimicrobial susceptibility of fecal escherichia coli isolates in dairy cows following systemic treatment with ceftiofur or penicillin.the objective of this longitudinal controlled trial was to determine the effect of systemic treatment with ceftiofur on antimicrobial susceptibility of fecal escherichia coli isolates in dairy cows. cows with metritis or interdigital necrobacillosis requiring systemic antimicrobial treatment were sequentially assigned to two treatment groups. the first group was treated with ceftiofur hydrochloride and the second with penicillin g procaine. untreated healthy control cows were selected for sampli ...201121381922
application of classification and regression tree (cart) analysis on the microflora of minced meat for classification according to reg. (ec) 2073/2005.in a retrospective study on the microbiology of minced meat from small food businesses supplying directly to the consumer, the relative contribution of meat supplier, meat species and outlet where meat was minced was assessed by "classification and regression tree" (cart) analysis. samples (n=888) originated from 129 outlets of a single supermarket chain. sampling units were 4-5 packs (pork, beef, and mixed pork-beef). total aerobic counts (tacs) were 5.3-¦1.0 log cfu/g. in 75.6% of samples, e. ...201121388752
modification of the bax system pcr assay for detecting salmonella in beef, produce, and soy protein isolate. performance tested method 100201.the bax system pcr assay for salmonella detection in foods was previously validated as aoac research institute (ri) performance tested method (ptm) 100201. new studies were conducted on beef and produce using the same media and protocol currently approved for the bax system pcr assay for e. coli o157:h7 multiplex (mp). additionally, soy protein isolate was tested for matrix extension using the u.s. food and drug administration-bacteriological analytical manual (fda-bam) enrichment protocols. the ...201121391494
evaluating the prevalence and genetic diversity of adenovirus and polyomavirus in bovine waste for microbial source tracking.this study evaluated and compared the occurrence, concentrations, and genetic diversity of bovine polyomavirus (bpyv) and bovine adenovirus (badv) in manure and feces samples for the purpose of determining which of these two viruses is more suitable for bovine fecal indication and microbial source tracking. the comparability and correlation between concentrations/prevalence of these viruses and bacterial fecal indicators (cow-associated bacteroidetes, escherichia coli and enterococci) in manure ...201121394527
bacterial survival studies to assess the efficacy of static pile composting and above ground burial for disposal of bovine carcases.to assess the survival of bacteria during two alternative means of cattle carcase disposal in windrows: static pile composting (spc) and above ground burial in soil (agb), under temperate climate conditions on agricultural land, compared to surface disposal as the control method.201121395951
genotypic-phenotypic discrepancies between antibiotic resistance characteristics of escherichia coli isolates from calves in management settings with high and low antibiotic use.we hypothesized that bacterial populations growing in the absence of antibiotics will accumulate more resistance gene mutations than bacterial populations growing in the presence of antibiotics. if this is so, the prevalence of dysfunctional resistance genes (resistance pseudogenes) could provide a measure of the level of antibiotic exposure present in a given environment. as a proof-of-concept test, we assayed field strains of escherichia coli for their resistance genotypes using a resistance g ...201121421795
protection of aquo/hydroxocobalamin from reduced glutathione by a b12 trafficking chaperone.we identified a bovine b(12) trafficking chaperone bcblc in bos taurus that showed 88% amino acid sequence identity with a human homologue. the protein bcblc was purified from e. coli by over-expression of the encoding gene. bcblc bound cyanocobalamin (cncbl), methylcobalamin (mecbl) and adenosylcobalamin (adocbl) in the base-off states and eliminated the upper axial ligands forming aquo/hydroxocobalamin (oh(2)/ohcbl) under aerobic conditions. a transition of oh(2)/ohcbl was induced upon binding ...201121429294
phospholemman (fxyd1) raises the affinity of the human a1ß1 isoform of na,k-atpase for na ions.the human a(1)/his(10)-ß(1) isoform of the na,k-atpase has been expressed in pichia pastoris, solubilized in n-dodecyl-ß-maltoside, and purified by metal chelate chromatography. the a(1)ß(1) complex spontaneously associates in vitro with the detergent-solubilized purified human fxyd1 (phospholemman) expressed in escherichia coli. it has been confirmed that fxyd1 spontaneously associates in vitro with the a(1)/his(10)-ß(1) complex and stabilizes it in an active mode. the functional properties of ...201121449573
a novel two-stage tandem mass spectrometry approach and scoring scheme for the identification of o-glcnac modified peptides.the modification of serine and threonine residues in proteins by a single n-acetylglucosamine (o-glcnac) residue is an emerging post-translational modification (ptm) with broad biological implications. however, the systematic or large-scale analysis of this ptm is hampered by several factors, including low stoichiometry and the lability of the o-glycosidic bond during tandem mass spectrometry. using a library of 72 synthetic glycopeptides, we developed a two-stage tandem ms approach consisting o ...201121472528
a prebiotic, celmanax™, decreases escherichia coli o157:h7 colonization of bovine cells and feed-associated cytotoxicity in vitro.abstract:201121473767
real-time quantification of mcra, pmoa for methanogen, methanotroph estimations during composting.composting is the controlled biological decomposition of organic matter by microorganisms during predominantly aerobic conditions. it is being increasingly adopted due to its benefits in nutrient recycling, soil reclamation, and urban land use. however, it poses an environmental concern related to its contribution to greenhouse gas production. during composting, activities of methanogenic and methanotrophic communities influence the net methane (ch4) release into the atmosphere. using quantitati ...201121488508
transcriptional regulators of the gad acid stress island are carried by effector protein-encoding prophages and indirectly control type iii secretion in enterohemorrhagic escherichia coli o157:h7.type iii secretion (t3s) plays a pivotal role in the colonization of ruminant hosts by enterohemorrhagic escherichia coli (ehec). the t3s system translocates effector proteins into host cells to promote bacterial attachment and persistence. the repertoire and variation in prophage regions underpins differences in the pathogenesis and epidemiology of ehec strains. in this study, we have used a collection of deletions in cryptic prophages and ehec o157 o-islands to screen for novel regulators of t ...201121492263
assessment of an extraction protocol to detect the major mastitis-causing pathogens in bovine milk.despite all efforts to control its spread, mastitis remains the most costly disease for dairy farmers worldwide. one key component of better control of this disease is identification of the causative bacterial agent during udder infections in cows. mastitis is complex, however, given the diversity of pathogens that must be identified. development of a rapid and efficient bacterial species identification tool is thus necessary. this study was conducted to demonstrate the feasibility of bacterial ...201121524507
effects of differential supplementation of fatty acids during the peripartum and breeding periods of holstein cows: ii. neutrophil fatty acids and function, and acute phase proteins.the objectives were to evaluate the effects of differential supplementation of ca salts (cs) of fatty acids (fa) on plasma acute phase proteins and both fa composition and function (i.e., activity and cytokine production) of neutrophils, during the peripartum and breeding periods. holstein cows were assigned randomly to receive either cs of palm (po) or safflower (so) oils from 30 d prepartum until 35 d postpartum (dpp) and cs of po or fish oil (fo) from 35 to 160 dpp. supplementation of cs of f ...201121524518
technical note: validation of candidate reference genes for normalization of quantitative pcr in bovine mammary epithelial cells responding to inflammatory stimuli.mammary epithelial cells (mec) participate in the first line of defense of the mammary gland to invading pathogens. in vitro culture of mec is widely used as a model to study the capacity of these cells to sense and respond to mastitis-causing bacteria. analysis of gene expression by quantitative pcr (qpcr) following exposure to bacteria or bacterial constituents is a powerful tool to assess responses of mec to pathogens. although internal standards such as reference genes are required for qpcr ...201121524534
bovine leukemia virus infection in dairy cattle: effect on serological response to immunization against j5 escherichia coli bacterin.thirteen bovine leukemia virus- (blv-) negative and 22 blv-positive holstein cows were immunized with j5 escherichia coli bacterin at dry off, three weeks before calving, during the second week after calving, and three weeks after the third immunization. serum was collected before the initial immunization, immediately before the third and fourth immunizations, and 21 days after the fourth immunization. anti-j5 e. coli igm, igg1, and igg2 titers were determined by elisa. anti-j5 e. coli igm titer ...201121547233
effect of repeated irrigation with water containing varying levels of total organic carbon on the persistence of escherichia coli o157:h7 on baby spinach.the california lettuce and leafy greens industry has adopted the leafy greens marketing agreement (lgma), which allows for 126 most-probable-number (mpn) escherichia coli per 100 ml in irrigation water. repeat irrigation of baby spinach plants with water containing e. coli o157:h7 and different levels of total organic carbon (toc) was used to determine the epiphytic survival of e. coli o157:h7. three irrigation treatments (0 ppm of toc, 12 or 15 ppm of toc, and 120 or 150 ppm of toc) were prepar ...201121549040
prevalence of salmonella enterica, listeria monocytogenes, and escherichia coli virulence factors in bulk tank milk and in-line filters from u.s. dairies.the zoonotic bacteria salmonella enterica, listeria monocytogenes, and escherichia coli are known to infect dairy cows while not always causing clinical signs of disease. these pathogens are sometimes found in raw milk, and human disease outbreaks due to these organisms have been associated with the consumption of raw milk or raw milk products. bulk tank milk (btm) samples (536) and in-line milk filters (519) collected from dairy farms across the united states during the national animal health m ...201121549046
antibacterial peptides derived from caprine whey proteins, by digestion with human gastrointestinal juice.peptides in caprine whey were identified after in vitro digestion with human gastrointestinal enzymes in order to determine their antibacterial effect. the digestion was performed in two continuing steps using human gastric juice (ph 2·5) and human duodenal juice (ph 8) at 37°c. after digestion the hydrolysate was fractionated and 106 peptides were identified. from these results, twenty-two peptides, located in the protein molecules, were synthesised and antibacterial activity examined. strong a ...201121554806
biomimetic anchors for antifouling and antibacterial polymer brushes on stainless steel.barnacle cement (bc) was beneficially applied on stainless steel (ss) to serve as the initiator anchor for surface-initiated polymerization. the amine and hydroxyl moieties of barnacle cement reacted with 2-bromoisobutyryl bromide to provide the alkyl halide initiator for the surface-initiated atom transfer radical polymerization (atrp) of 2-hydroxyethyl methacrylate (hema). the hydroxyl groups of hema polymer (phema) were then converted to carboxyl groups for coupling of chitosan (cs) to impart ...201121563843
stx genotype and molecular epidemiological analyses of shiga toxin-producing escherichia coli o157:h7/h- in human and cattle isolates.the relationship between human diseases caused by infection with shiga toxin (stx)-producing escherichia coli (stec) o157 strains and o157 strains isolated from cattle was investigated in an area where stockbreeding is prolific. for this purpose, the stx genotypes, the molecular epidemiological characteristics of 268 stec o157 strains including 211 human-origin strains and 57 cattle-origin strains, and clinical manifestations of 210 stec-infected people were analyzed. of 211 human-origin strains ...201121573816
changes in thermal nociceptive responses in dairy cows following experimentally induced escherichia coli mastitis.abstract: background: mastitis is a high incidence disease in dairy cows. the acute stage is considered painful and inflammation can lead to hyperalgesia and thereby contribute to decreased welfare. the aim of this study was to examine changes in nociceptive responses toward cutaneous nociceptive laser stimulation (nls) in dairy cows with experimentally induced escherichia coli mastitis, and correlate behavioral changes in nociceptive responses to clinical and paraclinical variables. methods: se ...201121592395
beta-35 is a transferrin-derived inhibitor of angiogenesis and tumor growth.an angiogenesis inhibitor named beta-35 has been identified and purified from the conditioned medium of mouse pancreatic β cells tumor cells. beta-35 has a molecular weight of 35kda and inhibits dna synthesis of bovine capillary endothelial cells at a half-maximal concentration of approximately 5nm. it shows anti-angiogenic activity in the chick embryo chorioallantoic membrane at a dose of about 1μg/embryo. amino acid microsequencing and mass spectrometric analysis of the purified protein demons ...201121605550
potential public health significance of faecal contamination and multidrug-resistant escherichia coli and salmonella serotypes in a lake in india.objective: to assess the prevalence of faecal coliform bacteria and multiple drug resistance among escherichia coli and salmonella serotypes from vembanadu lake. study design: systematic microbiological testing. methods: monthly collection of water samples were made from ten stations on the southern and northern parts of a salt water regulator constructed in vembanadu lake in order to prevent incursion of seawater during certain periods of the year. density of faecal colifrom bacteria was estima ...201121620427
a tricistronic human adrenodoxin reductase-adrenodoxin-cytochrome p450 27a1 vector system for substrate hydroxylation in escherichia coli.cytochrome p450 (p450) 27a1 catalyzes 27-hydroxylation of cholesterol and 25-hydroxylation of vitamin d(3), serving as an important component for the maintenance of lipid homeostasis. in eukaryotic cells p450 27a1 is a membrane-bound protein located on the inner mitochondrial membrane and requires two auxiliary reduction partners, adrenodoxin (adx) and nadph-adrenodoxin reductase (adr), for catalysis in the bile acid biosynthesis pathway. a strategy was developed for the functional coexpression ...201121621619
comparison of virulence gene profiles of escherichia coli isolates from sows with coliform mastitis and healthy sows.coliform mastitis (cm) is not only a serious economical and animal welfare touching problem in dairy cattle, but also in sows after farrowing. due to this disease, the essential adequate supply with colostrum for the growth and the health of the piglets is not ensured. besides other influencing factors, escherichia (e.) coli is of great importance as a causative agent of this multifactorial disease. in this study, e. coli isolates from milk samples of healthy and cm-affected sows were examined f ...201121621931
study of polymorphisms in tir, eae and tccp2 genes in enterohaemorrhagic and enteropathogenic escherichia coli of serogroup o26.abstract: background: enteropathogenic (epec) and enterohaemorrhagic (ehec) escherichia coli are responsible for food poisoning (enteritis and enterotoxaemia) in humans in developed countries. cattle are considered to be an important reservoir of ehec and epec strains for humans. moreover, some of the strains, belonging to the o26, o111, o118 serogroups, for example, are also responsible for digestive disorders in calves. the translocated intimin receptor (tir), the intimin (eae) and the tir-cyt ...201121624114
mouldy feed, mycotoxins and shiga toxin - producing escherichia coli colonization associated with jejunal hemorrhage syndrome in beef cattle.abstract: background: both o157 and non-o157 shiga toxin - producing escherichia coli (stecs) cause serious human disease outbreaks through the consumption of contaminated foods. cattle are considered the main reservoir but it is unclear how stecs affect mature animals. neonatal calves are the susceptible age class for stec infections causing severe enteritis. in an earlier study, we determined that mycotoxins and stecs were part of the disease complex for dairy cattle with jejunal hemorrhage sy ...201121639911
etiology and antimicrobial susceptibility of udder pathogens from cases of subclinical mastitis in dairy cows in sweden.abstract: background: a nationwide survey on the microbial etiology of cases of subclinical mastitis in dairy cows was carried out on dairy farms in sweden. the aim was to investigate the microbial panorama and the occurrence of acquired antimicrobial resistance. moreover, differences between newly infected cows and chronically infected cows were investigated. methods: in total, 583 quarter milk samples were collected from 583 dairy cows at 226 dairy farms from february 2008 to february 2009. th ...201121649936
[expression and antiviral assay of bovine interferon-gamma].bovine interferon-gamma (boifn-gamma) gene was amplified by reverse transcription polymerase chain reaction (rt-pcr) from total rna of bovine spleen lymphocytes stimulated with cona. the products of rt-pcr were cloned into pvax1 vector, positive recombinant clone was identified by restriction enzyme digestion and sequencing. the recombinant plasmid pvax1-bolfn-gamma was transfected into cos-7 cells mediated by lipofectine, indirect immunofluorescent assay analysis confirmed that rboifn-gamma was ...201121650053
effects of sodium bisulfate on the bacterial population structure of dairy cow waste.to determine the effects of sodium bisulfate (sbs) on the bacterial populations in cattle waste.201121651680
[prokaryotic expression of recombinant bovine il-4 and development of monoclonal antibodies against bovine il-4].to express recombinant bovine il-4 (rboil-4) in escherichia coli and prepare monoclonal antibody (mab) against rboil-4.201121651868
naturally colonized beef cattle populations fed combinations of yeast culture and an ionophore in finishing diets containing dried distiller's grains with solubles had similar fecal shedding of escherichia coli o157:h7.beef steers (n = 252) were used to evaluate the effects of dietary supplement on fecal shedding of escherichia coli o157:h7. seven pens of 9 steers (63 steers per treatment) were fed diets supplemented with or without yeast culture (yc) or monensin (mon) and their combination (yc × mon). yc and mon were offered at 2.8 g/kg and 33 mg/kg of dry matter intake, respectively. environmental sponge samples (from each pen floor, feed bunk, and water trough) were collected on day 0. rectal fecal grab sa ...201121669067
structures of glycosylated mammalian glutaminyl cyclases reveal conformational variability near the active center.formation of n-terminal pyroglutamate (pglu or pe) from glutaminyl or glutamyl precursors is catalyzed by glutaminyl cyclases (qc). as the formation of pglu-amyloid has been linked with alzheimer's disease, inhibitors of qcs are currently the subject of intense development. here, we report three crystal structures of n-glycosylated mammalian qc from humans (hqc) and mice (mqc). whereas the overall structures of the enzymes are similar to those reported previously, two surface loops in the neighb ...201121671571
endotoxin-free purification for the isolation of bovine viral diarrhoea virus e2 protein from insoluble inclusion body aggregates.abstract: background: protein expression in escherichia coli may result in the recombinant protein being expressed as insoluble inclusion bodies. in addition, proteins purified from e. coli contain endotoxins which need to be removed for in vivo applications. the structural protein, e2, from bovine viral diarrhoea virus (bvdv) is a major immunogenic determinant, and is an ideal candidate as a subunit vaccine. the e2 protein contains 17 cysteine residues creating difficulties in e. coli expressio ...201121787435
strategies for bacterial expression of protein-peptide complexes: application to solubilization of papillomavirus e6.e6 is a small oncoprotein involved in tumorigenesis induced by papillomaviruses (pvs). e6 often recognizes its cellular targets by binding to short motifs presenting the consensus lxxll. e6 proteins have long resisted structural analysis. we found that bovine papillomavirus type 1 (bpv1) e6 binds the n-terminal lxxll motif of the cellular protein paxillin with significantly higher affinity as compared to other e6/peptide interactions. although recombinant bpv1 e6 was poorly soluble in the free s ...201121777678
selection and expression of recombinant single domain antibodies from a hyper-immunized library against the hapten azoxystrobin.three v(h)hs against the model hapten, azoxystrobin (mw 403) were isolated from a hyper-immunized phage-displayed v(h)h library. this library was constructed by isolating the v(h)h-coding genes from the lymphocytes collected from a llama glama that was immunized with azoxystrobin conjugated to bovine serum albumin (bsa). six rounds of panning were performed against azoxystrobin conjugated to either ovalbumin (ova) or rabbit serum albumin (rsa) to enrich clones containing v(h)hs specific to the h ...201121777591
colostrum and amniotic fluid from different species exhibit similar immunomodulating effects in bacterium-stimulated dendritic cells.the fetus and newborn are immunologically immature. bioactive compounds in amniotic fluid (af) and maternal milk therefore play a key role in the immunological development of the infant intestine. we hypothesized that colostrum and af exert similar immunomodulatory effects on the developing immune system. hence, bone marrow-derived murine dendritic cells (bmdcs) were co-incubated with clostridium perfringens a or escherichia coli nissle 1917 and porcine, bovine, or human af, colostrum/milk whey ...201121777143
both the c-terminal polylysine region and the farnesylation of k-rasb are important for its specific interaction with calmodulin.ras protein, as one of intracellular signal switches, plays various roles in several cell activities such as differentiation and proliferation. there is considerable evidence showing that calmodulin (cam) binds to k-rasb and dissociates k-rasb from membrane and that the inactivation of cam is able to induce k-rasb activation. however, the mechanism for the interaction of cam with k-rasb is not well understood.201121750741
epidemiology of extended spectrum beta-lactamase e. coli (ctx-m-15) on a commercial dairy farm.the epidemiology of an extended spectrum beta-lactamase escherichia coli (ctx-m-15) was observed and described on a commercial dairy farm located in the united kingdom. during 2008 longitudinal sampling of faecal pat samples from different cattle groups comprising milking and non-milking cows, calving cows, calves, and the environment was carried out. the proportion of ctx-m-15 e. coli positive samples was significantly (p<0.0.01) higher in milking cows (30.3%, ci(95%) 26.8; 33.8) than in the he ...201121840142
[microbiological quality of the water used in a random sample from dairy farms in c+¦rdoba, argentina].bacteriological contamination of well water and wash water in a random sample obtained from 50 farms from villa mar+¡a (c+¦rdoba) dairy area, argentina, was evaluated during a visit in 2007. forty six percent and 24 % of farms showed an aerobic mesophilic bacteria count higher than 500 cfu/ml in wash water and well water, respectively. escherichia coli was isolated in 20 % of samples from both sources. pseudomonas aeruginosa showed high frequency of isolation in well water (36 %) and wash water ...201121731973
internalization of escherichia coli o157:h7 by bovine rectal epithelial cells.escherichia coli o157:h7 (o157) causes human diarrheal disease and healthy cattle are its primary reservoir. o157 colonize the bovine epithelial mucosa at the recto-anal junction (raj). previous studies show that o157 at this site are not eliminated by aggressive interventions including applications of o157-specific lytic bacteriophages and other bactericidal agents. we hypothesize that some o157 at the raj mucosa are protected from these killing agents by host cell internalization. to test this ...201121687423
evaluation of escherichia coli o157:h7 translocation and decontamination for beef vacuum-packaged subprimals destined for nonintact use.the translocation of escherichia coli o157:h7 as well as the impact of water washing and partial or complete surface trimming as possible pathogen reduction strategies were evaluated for vacuum-packaged beef subprimals destined for nonintact use. cap-on and cap-off beef top sirloin butts were inoculated with two levels of e. coli o157:h7: a high-inoculum level of approximately 10(4) cfu/cm(2) and a low-inoculum level of approximately 10(2) cfu/cm(2). following inoculation, the subprimals were va ...201121740705
the use of a radiotelemetric ruminal bolus to detect body temperature changes in lactating dairy cattle.the objective of this study was to validate the efficacy of a radiotelemetric bolus (rtb) to detect changes in ruminal temperature resulting from (1) systemic illnesses that are associated with febrile responses and (2) subacute ruminal acidosis (sara). eight rumen-fistulated, lactating holstein cows (586±37 kg of body weight, 106±18 d in milk) were used in a replicated 4 × 4 latin square design with a 2 × 2 factorial arrangement. each period consisted of 21 d. the factors were 2 diets, a mo ...201121700044
vaccination with type iii secreted proteins leads to decreased shedding in calves after experimental infection with escherichia coli o157.escherichia coli o157:h7 remains a threat to humans via cattle-derived fecal contamination of food and water. preharvest intervention strategies represent a means of reducing the pathogen burden before harvest. in this study, the efficacy of a commercially produced type iii secreted protein (ttsp) vaccine was evaluated with the use of a commingled experimental calf infection model (30 placebo-treated animals and 30 vaccinates). the calves were vaccinated on days 0, 21, and 42 and infected with 1 ...201121731179
immediate and carryover effects of gram-negative and gram-positive toxin-induced mastitis on follicular function in dairy cows.this study compared immediate and carryover effects of mastitis induced by gram-negative endotoxin (e. coli lps) and gram-positive exosecretions (staph. aureus ex.) on preovulatory follicle function. synchronized, uninfected cyclic lactating holstein cows were treated with pgf(2+¦) on day 6 of the cycle and 36 h later, a dose of either e. coli lps (n = 8), s. aureus ex. (n = 10), or saline (n = 9) was administered into the mammary gland. follicular fluids and granulosa cells were aspirated 6 h l ...201121705051
a systematic review of vaccinations to reduce the shedding of escherichia coli o157 in the faeces of domestic ruminants.the objective of this study was to conduct a systematic review and meta-analysis of the published literature to evaluate the efficacy of vaccines in reducing faecal shedding of escherichia coli o157 in ruminants. a systematic search of eight databases and land-grant university research reports using an algorithm adapted from a previous systematic review of pre-harvest interventions against e. coli o157 was conducted to locate all reports of in vivo trials of e. coli o157 vaccines in ruminants pu ...201121824378
comparative expression profiling of e. coli and s. aureus inoculated primary mammary gland cells sampled from cows with different genetic predispositions for somatic cell score.during the past ten years many quantitative trait loci (qtl) affecting mastitis incidence and mastitis related traits like somatic cell score (scs) were identified in cattle. however, little is known about the molecular architecture of qtl affecting mastitis susceptibility and the underlying physiological mechanisms and genes causing mastitis susceptibility. here, a genome-wide expression analysis was conducted to analyze molecular mechanisms of mastitis susceptibility that are affected by a spe ...201121702919
the preference for water nipples vs. water bowls in dairy goats.previous studies have reported that the design of the water dispensers can influence the water intake in farm animals. horses and dairy cows seem to prefer to drink from an open surface whereas sheep and pigs apparently prefer water nipples, probably because of the worse water quality in water bowls. the aim of the present study was to examine the preference of dairy goats for water nipples or water bowls.201121939533
burger preparation: what consumers say and do in the home.ground beef has been linked to outbreaks of pathogenic bacteria like escherichia coli o157:h7 and salmonella. consumers may be exposed to foodborne illness through unsafe preparation of ground beef. video footage of 199 volunteers in northern california preparing hamburgers and salad was analyzed for compliance with u.s. department of agriculture recommendations and for violations of the u.s. food and drug administration's food code 2009. a questionnaire about consumer attitudes and knowledge a ...201122004819
bovine recto-anal junction squamous epithelial (rse) cell adhesion assay for studying escherichia coli o157 adherence.to develop a new adherence assay, using cattle recto-anal junction squamous epithelial (rse) cells, for evaluating bacterial adherence to cells of bovine origin.201121883734
[Colonization of the teat skin and the teat canal by mastitis pathogens in dairy cattle].Objective: The teat canal of lactating dairy cattle is the first barrier against mastitis pathogens invading the teat cistern and the udder lumen. However, in several studies it could be shown that the teat skin and the teat canal epithelium are colonized by mainly staphylococci as well as by other pathogenic or facultative pathogenic micro-organisms. The aim of the study was to evaluate the pathogen pressure on the bovine mammary gland and to establish data on the prevalence of relevant micro- ...201122138767
multielement analysis of micro-volume biological samples by icp-ms with highly efficient sample introduction system.a method for multielement analysis of micro-volume biological sample by inductively coupled plasma mass spectrometry (icp-ms) with a highly efficient sample introduction system was presented. the sample introduction system was the combination of (1) an inert loop injection unit and (2) a high performance concentric nebulizer (hpcn) coupled with a temperature controllable cyclone chamber. the loop injection unit could introduce 20 μl samples into the carrier liquid flow of 10 μl min(-1) producing ...201122099643
Association Between Antimicrobial Resistance in Escherichia coli Isolates from Food Animals and Blood Stream Isolates from Humans in Europe: An Ecological Study.Abstract Background: In addition to medical antimicrobial usage, the use of antimicrobials in food animals contributes to the occurrence of resistance among some bacterial species isolated from infections in humans. Recently, several studies have indicated that a large proportion of Escherichia coli causing infections in humans, especially those resistant to antimicrobials, have an animal origin. Methods: We analyzed the correlation between the prevalence of antimicrobial resistance in E. co ...201121883007
Heparan sulfate proteoglycans mediate the angiogenic activity of the vascular endothelial growth factor receptor-2 agonist gremlin.Heparan sulfate proteoglycans (HSPGs) modulate the interaction of proangiogenic heparin-binding vascular endothelial growth factors (VEGFs) with signaling VEGF receptor-2 (VEGFR2) and neuropilin coreceptors in endothelial cells (ECs). The bone morphogenic protein antagonist gremlin is a proangiogenic ligand of VEGFR2, distinct from canonical VEGFs. Here we investigated the role of HSPGs in VEGFR2 interaction, signaling, and proangiogenic capacity of gremlin in ECs.201121921258
serotypes and virulence profiles of non-o157 shiga toxin-producing escherichia coli isolates from bovine farms.non-o157 shiga toxin-producing escherichia coli (stec) strains are clinically significant food-borne pathogens. however, there is a dearth of information on serotype prevalence and virulence gene distribution, data essential for the development of public health protection monitoring and control activities for the meat and dairy industries. thus, the objective of this study was to examine the prevalence of non-o157 stec on beef and dairy farms and to characterize the isolates in terms of serotype ...201122003024
Selective reduction of the pathogenic load of cow manure in an industrial-scale continuous-feeding vermireactor.Vermicomposting is a suitable technology for processing different wastes, to produce a valuable end product (vermicompost). However, the pathogenic load of the waste must be greatly reduced in order to prevent risks to human health. Although Eisenia andrei may reduce the levels of several pathogens, the feasibility of vermicomposting, with regard to pathogen reduction, has not been tested on an industrial scale. This work studied whether vermicomposting in a continuous feeding vermireactor, is a ...201121875788
Effect of Escherichia coli lipopolysaccharide on u-PA activity and u-PA and u-PAR RNA expression in a bovine mammary epithelial cell line.It is well known that the plasminogen-activating (PA) system plays a key role in the bovine mammary gland during tissue remodelling. However, the modulation of the PA cascade after bacterial infections needs to be elucidated. This study examined the effects of Escherichia coli lipopolysaccharide (LPS) on cell viability, the modulation of cell-associated u-PA activity, and the regulation of u-PA and u-PA receptor (u-PAR) RNA expression using the BME-UV1 bovine mammary epithelial cell line. LPS di ...201122103977
Carbon nanoparticles as detection labels in antibody microarrays. Detection of genes encoding virulence factors in Shiga toxin-producing Escherichia coli.The present study demonstrates that carbon nanoparticles (CNPs) can be used as labels in microarrays. CNPs were used in nucleic acid microarray immunoassays (NAMIAs) for the detection of different Shiga toxin-producing Escherichia coli (STEC) virulence factors: four genes specific for STEC (vt1, vt2, eae, and ehxA) and the gene for E. coli 16S (hui). Optimization was performed using a Box-Behnken design, and the limit of detection for each virulence factor was established. Finally, this NAMIA us ...201121936575
rapid identification of bovine mastitis pathogens by high-resolution melt analysis of 16s rdna sequences.accurate identification of mastitis pathogens is often compromised when using conventional culture-based methods. here, we report a novel, rapid assay tested for speciation of bacterial mastitis pathogens using high-resolution melt analysis (hrma) of 16s rdna sequences. real-time pcr amplification of 16s rrna gene fragment, spanning the variable region v5 and v6 was performed with a resulting amplicon of 290bp. first, a library was generated of melt curves of 9 common pathogens that are implicat ...201121944716
enhanced sensitivity for selected reaction monitoring mass spectrometry-based targeted proteomics using a dual stage electrodynamic ion funnel interface.selected reaction monitoring mass spectrometry (srm-ms) is playing an increasing role in quantitative proteomics and biomarker discovery studies as a method for high throughput candidate quantification and verification. although srm-ms offers advantages in sensitivity and quantification compared with other ms-based techniques, current srm technologies are still challenged by detection and quantification of low abundance proteins (e.g. present at ∼10 ng/ml or lower levels in blood plasma). here w ...201120410378
identification of papain-like cysteine proteases from the bovine piroplasm babesia bigemina and evolutionary relationship of piroplasms c1 family of cysteine proteases.papain-like cysteine proteases have been shown to have essential roles in parasitic protozoa and are under study as promising drug targets. five genes were identified by sequence similarity search to be homologous to the cysteine protease family in the ongoing babesia bigemina genome sequencing project database and were compared with the annotated genes from the complete bovine piroplasm genomes of babesia bovis, theileria annulata, and theileria parva. multiple genome alignments and sequence an ...201120655912
use of model super-shedders to define the role of pen floor and hide contamination in the transmission of escherichia coli o157:h7.super-shedders, cattle shedding at least 10(4) cfu of escherichia coli o157:h7 per gram of feces, increase the risks of contaminating the food chain and disseminating the organism through cattle populations. because detecting super-shedders in cattle populations is laborious and time-consuming, a study was conducted to evaluate the role of hide and pen-floor contamination by model super shedders (mss) in transmission of e. coli o157:h7. steers (n = 48) negative for e. coli o157:h7 were allocated ...201120852081
phylogeny, virulence factors and antimicrobial susceptibility of escherichia coli isolated in clinical bovine mastitis.the aim of this study was to identify specific phylogeny groups, virulence genes or antimicrobial resistance traits of escherichia coli isolated in bovine mastitis associated to clinical signs, persistence of intramammary infection in the quarter and recovery from mastitis. a total of 154 e. coli isolates from bovine clinical mastitis, 144 from the acute stage and 10 from follow-up samples 3 weeks later, originating from 144 cows in 65 dairy herds in southern finland were investigated. phylogeny ...201120729012
genetic variations in shiga toxin-producing abilities of bovine and human escherichia coli o157:h7.cattle are a primary reservoir of escherichia coli o157:h7, a major foodborne pathogen. the organism causes haemorrhagic colitis which can lead to serious complications, including haemolytic-uraemic syndrome. although e. coli o157:h7 is widely prevalent in cattle and cattle environments, the number of human cases remain relatively low, suggesting possible strain diversity and differences in virulence between human and bovine strains. shiga toxins, stx1 and stx2, are the major virulence factors. ...201120819202
conjugation and fluorescence quenching between bovine serum albumin and l-cysteine capped cdse/cds quantum dots.water-soluble, biological-compatible, and excellent fluorescent cdse/cds quantum dots (qds) with l-cysteine as capping agent were synthesized in aqueous medium. fluorescence (fl) spectra, absorption spectra, and transmission electron microscopy (tem) were employed to investigate the quality of the products. the interactions between qds and bovine serum albumin (bsa) were studied by absorption and fl titration experiments. with addition of qds, the fl intensity of bsa was significantly quenched w ...201121121888
determination of ceftiofur derivatives in serum, endometrial tissue, and lochia in puerperal dairy cows after subcutaneous administration of ceftiofur crystalline free acid.puerperal uterine infections are often associated with decreased reproductive performance in dairy cows. routine treatment protocols include the systemic administration of antibiotics. antibiotic drugs, however, should be administered daily over at least 5 d. the objective of this study was to determine concentrations of ceftiofur derivatives in serum, endometrial tissue, and lochia after subcutaneous administration of ceftiofur crystalline free acid in 6 clinically healthy puerperal dairy cows ...201121183038
population structure of rumen escherichia coli associated with subacute ruminal acidosis (sara) in dairy cattle.previous studies indicated that only subacute ruminal acidosis (sara), induced by feeding a high-grain diet, is associated with an inflammatory response and increased abundance of escherichia coli in the rumen. we hypothesized that ruminal e. coli in grain pellet-induced sara carried virulence factors that potentially contribute to the immune activation during sara. one hundred twenty-nine e. coli isolates were cultured from the rumens of 8 cows (4 animals per treatment) in which sara had been n ...201121183045
intracellular fate of strains of escherichia coli isolated from dairy cows with acute or chronic mastitis.research on mastitis in dairy cows caused by escherichia coli has reported the emergence of strains capable of inducing chronic mastitis and that these strains adhered to and internalized into bovine mammary epithelial cells better than strains of e. coli isolated from acute mastitis. to understand mechanisms and strategies used by chronic e. coli strains to survive intracellularly internalization studies using bovine mammary epithelial cells treated with inhibitors of caveolae-mediated endocyto ...201121207146
a novel organic-inorganic hybrid monolith for trypsin immobilization.in proteomics, attention has focused on various immobilized enzyme reactors (imers) for the realization of high throughput digestion. in this report, a novel organic-inorganic hybrid monolith based imer was prepared in a 100 μm i.d. capillary with 3-glycidoxypropyltrimethoxysilane (glymo) as the monomer and tetraethoxysilane (teos) as the crosslinker. trypsin immobilization was achieved via the reaction between vicinal diol groups, which were obtained from hydrolysis of epoxy groups, and the ami ...201121253871
lingual antimicrobial peptide and il-8 expression are oppositely regulated by the antagonistic effects of nf-κb p65 and c/ebpβ in mammary epithelial cells.pathogen contact induces quickly in mammary epithelial cells (mec) the expression of the proinflammatory cytokine il-8 and delayed that of the bactericidal β-defensin lap. both genes encoding these factors feature on their proximal promoter a composite nf-κb/cebp binding site. we compare here in mec the role of nf-κb and c/ebp factors in regulating basal and pathogen-induced expression of both genes from cattle. abrogating nf-κb binding to that site by introduction of a single point mutation blo ...201121255844
cdna, genomic sequence cloning and overexpression of the ribosomal protein s13 gene in the giant panda (ailuropoda melanoleuca).the cdna and the genomic sequence of ribosomal protein s13 (rps13) of the giant panda (ailuropoda melanoleuca) was cloned using reverse transcription-polymerase chain reaction (rt-pcr) and touchdown-pcr, respectively. these two sequences were sequenced and analyzed, and the cdna of the rps13 gene was overexpressed in escherichia coli bl21. we compared the nucleotide sequences of the coding region and the amino acid sequences with those of seven other mammalian species retrieved from genbank. the ...201121268784
long target droplet polymerase chain reaction with a microfluidic device for high-throughput detection of pathogenic bacteria at clinical sensitivity.in this article we present a long target droplet polymerase chain reaction (pcr) microsystem for the amplification of the 16s ribosomal rna gene. it is used for detecting gram-positive and gram-negative pathogens at high-throughput and is optimised for downstream species identification. the miniaturised device consists of three heating plates for denaturation, annealing and extension arranged to form a triangular prism. around this prism a fluoropolymeric tubing is coiled, which represents the r ...201121271358
prevalence of shiga toxin-encoding genes and risk factors among dairy farmers in gyeonggi province, korea.dairy farmers perform various types of work and are in direct contact with dairy cattle nearly every day. the purpose of this study was to assess the prevalence of shiga toxin-encoding genes (stx) among dairy farmers and to evaluate the relationship between stx and risk factors.201121231809
differentiation of fecal escherichia coli from human, livestock, and poultry sources by rep-pcr dna fingerprinting on the shellfish culture area of east china sea.the rep-pcr dna fingerprinting performed with rep, box a1r, and (gtg)(5) primers was investigated as a way to differentiate between human, livestock, and poultry sources of fecal pollution on the area of xiangshan bay, east china sea. of the three methods, the box-pcr dna fingerprints analyzed by jack-knife algorithm were revealed high rate of correct classification (rcc) with 91.30, 80.39, 89.39, 86.14, 93.24, 87.72, and 89.28% of human, cattle, swine, chicken, duck, sheep, and goose e. coli is ...201121279641
alkbh8-mediated formation of a novel diastereomeric pair of wobble nucleosides in mammalian trna.mammals have nine different homologues (alkbh1-9) of the escherichia coli dna repair demethylase alkb. alkbh2 is a genuine dna repair enzyme, but the in vivo function of the other alkbh proteins has remained elusive. it was recently shown that alkbh8 contains an additional transfer rna (trna) methyltransferase domain, which generates the wobble nucleoside 5-methoxycarbonylmethyluridine (mcm(5)u) from its precursor 5-carboxymethyluridine (cm(5)u). in this study, we report that (r)- and 5-methoxyc ...201121285950
insights into mucosal innate responses to escherichia coli o157 : h7 colonization of cattle by mathematical modelling of excretion dynamics.mathematical model-based statistical inference applied to within-host dynamics of infectious diseases can help dissect complex interactions between hosts and microbes. this work has applied advances in model-based inference to understand colonization of cattle by enterohaemorrhagic escherichia coli o157 : h7 at the terminal rectum. a mathematical model was developed based on niche replication and transition rates at this site. a nested-model comparison, applied to excretion curves from 25 calves ...201121849385
Arcanolysin is a cholesterol-dependent cytolysin of the human pathogen Arcanobacterium haemolyticum.Arcanobacterium haemolyticum is an emerging human pathogen that causes pharyngitis, wound infections, and a variety of occasional invasive diseases. Since its initial discovery in 1946, this Gram positive organism has been known to have hemolytic activity, yet no hemolysin has been previously reported. A. haemolyticum also displays variable hemolytic activity on laboratory blood agar that is dependent upon which species the blood is derived.201122029628
molecular analysis and recombinant expression of bovine neutrophil β-defensin 12 and its antimicrobial activity.to analyze molecular characteristics and antimicrobial activity of bovine neutrophil β-defensin 12 (bnbd12), pro-bnbd12 gene was amplified from chinese holstein cow using reverse transcription polymerase chain reaction. based on the sequence of mature bnbd12 and codon preference in e. coli, a modified mature bnbd12 cdna was synthesized, cloned into pet32a (+) vector and expressed in e. coli bl21 as a 26 kd fusion protein after isopropyl β-d-1-thiogalactopyranoside induction. the expressed mature ...201120349279
dairy cows with metritis: coxiella burnetii test results in uterine, blood and bulk milk samples.in cattle, coxiella burnetii infections are generally asymptomatic but can also be associated with reproductive disorders. metritis is considered as one of the symptoms of c. burnetii infections, but reliable information is lacking. therefore, information on the presence of c. burnetii in the uterine content of cows with metritis is important to increase our knowledge on this pathogen. in this study, the uterine content of 45 dairy cows with metritis belonging to 12 herds was tested for c. burne ...201120576366
antimicrobial activity of soy edible films incorporated with thyme and oregano essential oils on fresh ground beef patties.antibacterial activity of soy protein edible films (spef) incorporated with 1, 2, 3, 4 and 5% oregano (or) or thyme (th) essential oils was evaluated against escherichia coli, e. coli o157:h7, staphylococcus aureus, pseudomonas aeruginosa and lactobacillus plantarum by the inhibition zone test. effects of spef containing 5% or and th or a mixture of or+th (ort) were also tested on fresh ground beef during refrigerated storage (at 4 degrees c). or and th incorporated spef exhibited similar antiba ...201020580990
[transition of entheropathogenic and saprotrophic bacteria in the eco-niche cycle: animals-excrement-soil-plants-animals].the possibility of transition of saprotrophic and enteropathohenic bacterial populations following the chain of naturally related habitats--fodder-animal gastrointestinal tract (git)-animals excrement-soil-plants and again animals with a cyclic formation--has been investigated quantitatively. all bacteria used in the experiments have been shown to successfully overcome all the mechanical, physical-chemical, and biological barriers in the food chain and to come out into the environment with a qui ...201020583615
chemiluminescence of non-differentiated thp-1 promonocytes: developing an assay for screening anti-inflammatory milk proteins and peptides.human leukemic thp-1 promonocytes are widely used as a model for peripheral blood monocytes. however, superoxide production during respiratory burst (rb) of non-differentiated thp-1 (nd-thp-1) cells is very low. here we present a rapid and low-cost method for measuring the chemiluminescence (cl) of opsonized zymosan (oz) induced rb which allows detection of escherichia coli lipopolysaccharide (lps) induced priming of nd-thp-1 cells on the basis of cl reaction kinetics. maximum cl intensity obtai ...201020586113
cdna, genomic sequence cloning and overexpression of glyceraldehyde-3-phosphate dehydrogenase gene (gapdh) from the giant panda.gapdh (glyceraldehyde-3-phosphate dehydrogenase) is a key enzyme of the glycolytic pathway and it is related to the occurrence of some diseases. the cdna and the genomic sequence of gapdh were cloned successfully from the giant panda (ailuropoda melanoleuca) using the rt-pcr technology and touchdown-pcr, respectively. both sequences were analyzed preliminarily. the cdna of gapdh cloned from the giant panda is 1191 bp in size, contains an open reading frame of 1002 bp encoding 333 amino acids. th ...201020586183
cdna, genomic sequence cloning, and overexpression of eif1 from the giant panda (ailuropoda melanoleuca) and the black bear (ursus thibetanus mupinensis).eukaryotic initiation factor (eif) eif1 is a universally conserved translation factor that is involved in translation initiation site selection. the cdna and the genomic sequences of eif1 were cloned successfully from the giant panda (ailuropoda melanoleuca) and the black bear (ursus thibetanus mupinensis) using reverse transcription polymerase chain reaction (rt-pcr) technology and touchdown-polymerase chain reaction, respectively. the cdnas of the eif1 cloned from the giant panda and the black ...201020589574
kinetic and thermodynamic properties of two barley thioredoxin h isozymes, hvtrxh1 and hvtrxh2.barley thioredoxin h isozymes 1 (hvtrxh1) and barley thioredoxin h isozymes 2 (hvtrxh2) show distinct spatiotemporal distribution in germinating seeds. using a novel approach involving measurement of bidirectional electron transfer rates between escherichia coli thioredoxin, which exhibits redox-dependent fluorescence, and the barley isozymes, reaction kinetics and thermodynamic properties were readily determined. the reaction constants were approximately 60% higher for hvtrxh1 than hvtrxh2, whi ...201020594550
interaction studies of novel cell selective antimicrobial peptides with model membranes and e. coli atcc 11775.cationic antimicrobial peptides (camps) are novel candidates for drug development. here we describe design of six short and potent camps (sa-1 to sa-6) based on a minimalist template of 12 residues h+hhg+hh+hh+nh2 (where h: hydrophobic amino acid and +: charged hydrophilic amino acid). designed peptides exhibit good antibacterial activity in micro molar concentration range (1-32 mug/ml) and rapid clearance of gram-positive and gram-negative bacterial strains at concentrations higher than mic. fo ...201020599694
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