| enzymatic properties of aldh1l2, a mitochondrial 10-formyltetrahydrofolate dehydrogenase. | 10-formyltetrahydrofolate dehydrogenase (fdh, aldh1l1), an abundant cytosolic enzyme of folate metabolism, shares significant sequence similarity with enzymes of the aldehyde dehydrogenase (aldh) family. the enzyme converts 10-formyltetrahydrofolate (10-fthf) to tetrahydrofolate and co(2) in an nadp(+)-dependent manner. the mechanism of this reaction includes three consecutive steps with the final occurring in an aldh-homologous domain. we have recently identified a mitochondrial isoform of fdh ... | 2011 | 21238436 |
| structural basis for two-component system inhibition and pilus sensing by the auxiliary cpxp protein. | bacteria are equipped with two-component systems to cope with environmental changes, and auxiliary proteins provide response to additional stimuli. the cpx two-component system is the global modulator of cell envelope stress in gram-negative bacteria that integrates very different signals and consists of the kinase cpxa, the regulator cpxr, and the dual function auxiliary protein cpxp. cpxp both inhibits activation of cpxa and is indispensable for the quality control system of p pili that are cr ... | 2011 | 21239493 |
| successful treatment of sepsis-induced disseminated intravascular coagulation in a patient with idiopathic thrombocytopenic purpura using recombinant human soluble thrombomodulin. | disseminated intravascular coagulation (dic) may complicate a variety of disorders that contribute to mortality, particularly those related to bleeding. it is therefore very difficult to manage dic in patients with known bleeding disorders. we treated a 62-year-old woman with idiopathic thrombocytopenic purpura (itp) complicated with sepsis-induced dic. she had been diagnosed with itp 8 months prior to admission. laboratory tests showed an elevation of d-dimer and endotoxin, while pyelonephritis ... | 2011 | 21240494 |
| uafb is a serine-rich repeat adhesin of staphylococcus saprophyticus that mediates binding to fibronectin, fibrinogen and human uroepithelial cells. | staphylococcus saprophyticus is an important cause of urinary tract infection (uti), particularly among young women, and is second only to uropathogenic escherichia coli as the most frequent cause of uti. the molecular mechanisms of urinary tract colonization by s. saprophyticus remain poorly understood. we have identified a novel 6.84 kb plasmid-located adhesin-encoding gene in s. saprophyticus strain ms1146 which we have termed uro-adherence factor b (uafb). uafb is a glycosylated serin ... | 2011 | 21252279 |
| ecor phylogenetic analysis and virulence genotyping of avian pathogenic escherichia coli strains and escherichia coli isolates from commercial chicken carcasses in southern brazil. | escherichia coli strains designated as avian pathogenic e. coli (apec) are responsible for avian colibacillosis, an acute and largely systemic disease that promotes significant economic losses in poultry industry worldwide because of mortality increase, medication costs, and condemnation of carcasses. apec is a subgroup of extraintestinal pathogenic e. coli pathotype, which includes uropathogenic e. coli, neonatal meningitis e. coli, and septicemic e. coli. we isolated e. coli from commercial ch ... | 2011 | 21254888 |
| shiga toxin and its use in targeted cancer therapy and imaging. | shiga and the shiga-like toxins are related protein toxins produced by shigella dysenteriae and certain strains of escherichia coli. these toxins are composed of two non-covalently attached, modular parts: the a moiety (stxa) containing the enzymatically active a1 fragment, and the non-toxic, pentameric binding moiety (stxb). stx binds specifically to the glycosphingolipid globotriaosylceramide (gb3) at the surface of target cells and is then internalized by endocytosis. subsequently, in toxin-s ... | 2011 | 21255370 |
| a photopolymerized antimicrobial hydrogel coating derived from epsilon-poly-l-lysine. | hydrogels made from epsilon-poly-l-lysine-graft-methacrylamide (epl-ma) have been found to have impressive wide spectrum antimicrobial activity against both bacteria (specifically escherichia coli, pseudomonas aeruginosa, serratia marcescens and staphylococcus aureus) and fungi (specifically candida albicans and fusarium solani). the epl-ma hydrogel also possesses in vitro biocompatibility and epl-ma solution is relatively non-hemolytic: the concentration needed for onset of human red blood cell ... | 2011 | 21257199 |
| purification and characterization of cystathionine ß-synthase bearing a cobalt protoporphyrin. | human cystathionine ß-synthase (cbs), a pivotal enzyme in the metabolism of homocysteine, is a pyridoxal-5'-phosphate-dependent enzyme that also contains heme, a second cofactor whose function is still unclear. one strategy for elucidation of heme function is its replacement with different metalloporphyrins or with porphyrins containing different substituent groups. this paper describes a novel expression approach and purification of cobalt cbs (cocbs), which results in a high yield of fully act ... | 2011 | 21262193 |
| new ligation-independent cloning vectors compatible with a high-throughput platform for parallel construct expression evaluation using baculovirus-infected insect cells. | biomedical research has undergone a major shift in emphasis over the past decade from characterizing the genomes of organisms to characterizing their proteomes. the high-throughput approaches that were successfully applied to sequencing of genomes, such as miniaturization and automation, have been adapted for high-throughput cloning and protein production. high-throughput platforms allow for a multi-construct, multi-parallel approach to expression optimization and construct evaluation. we descri ... | 2011 | 21262364 |
| pharmacodynamics of razupenem (pz601) studied in an in vitro pharmacokinetic model of infection. | simulations of administration of razupenem at 1 g every 12 h by 1-h intravenous (i.v.) infusion were performed in an in vitro pharmacokinetic model of infection. the antibacterial effect of this razupenem dosing regimen against six strains of staphylococcus aureus (one methicillin-sensitive s. aureus [mssa] strain [mic, 0.015 µg/ml] and five methicillin-resistant s. aureus [mrsa] strains [mic range, 0.09 to 3 µg/ml]) and five strains of enterobacteriaceae (three escherichia coli strains [two con ... | 2011 | 21263054 |
| human alkb homologue 5 is a nuclear 2-oxoglutarate dependent oxygenase and a direct target of hypoxia-inducible factor 1a (hif-1a). | human 2-oxoglutarate oxygenases catalyse a range of biological oxidations including the demethylation of histone and nucleic acid substrates and the hydroxylation of proteins and small molecules. some of these processes are centrally involved in regulation of cellular responses to hypoxia. the alkbh proteins are a sub-family of 2og oxygenases that are defined by homology to the escherichia coli dna-methylation repair enzyme alkb. here we report evidence that alkbh5 is probably unique amongst the ... | 2011 | 21264265 |
| expression of human papillomavirus 16 e7ggg oncoprotein on n- and c-terminus of potato virus x coat protein in bacterial and plant cells. | the e7 oncoprotein from human papillomavirus type 16 (hpv16) is an attractive candidate for anti-cancer therapeutical vaccine development. in this study, we engineered different fusions of mutagenized coding sequence of e7 oncoprotein (e7ggg) with coat protein of potato virus x (pvx cp) both on 5'- and 3'-terminus of pvx cp and evaluated the influence of the length of linker (no linker, 4, 15aa) connecting pvx cp and e7ggg on their production. at first the expression in escherichia coli was cond ... | 2011 | 21266198 |
| 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 ... | 2011 | 21268784 |
| macrophages from ibd patients exhibit defective tumour necrosis factor-a secretion but otherwise normal or augmented pro-inflammatory responses to infection. | defects in macrophage function have been implicated in the establishment of crohn's disease (cd). however, the response of macrophages from cd patients to live bacteria, particularly mycobacterium avium subsp. paratuberculosis (map), has not been addressed. considering map has long been associated to cd, our objective was to assess whether macrophages from cd patients showed impaired inflammatory response to infection by map comparing to m. avium subsp. avium (ma) and other live intestinal comme ... | 2011 | 21269730 |
| bifidobacteria can protect from enteropathogenic infection through production of acetate. | the human gut is colonized with a wide variety of microorganisms, including species, such as those belonging to the bacterial genus bifidobacterium, that have beneficial effects on human physiology and pathology. among the most distinctive benefits of bifidobacteria are modulation of host defence responses and protection against infectious diseases. nevertheless, the molecular mechanisms underlying these effects have barely been elucidated. to investigate these mechanisms, we used mice associate ... | 2011 | 21270894 |
| distinct roles of protein disulfide isomerase and p5 sulfhydryl oxidoreductases in multiple pathways for oxidation of structurally diverse storage proteins in rice. | in the rice (oryza sativa) endosperm, storage proteins are synthesized on the rough endoplasmic reticulum (er), in which prolamins are sorted to protein bodies (pbs) called type-i pb (pb-i). protein disulfide isomerase (pdi) family oxidoreductase pdil2;3, an ortholog of human p5, contains a conserved structural disulfide in the redox-inactive thioredoxin-like (trx) domain and was efficiently targeted to the surface of pb-i in a redox active site-dependent manner, whereas pdil1;1, an ortholog of ... | 2011 | 21278127 |
| protein prenylation contributes to the effects of lps on efs-induced responses in human isolated bronchi. | rho/ras signalling pathways may play an important role in the mechanism of lps-induced inflammation and bronchoconstriction. in this study we investigated the effect of lps on the transmural contraction induced by efs on human isolated bronchi and examined a possible contribution of rho/ras signalling pathways by using geranylgeranyl (ggpp) and farnesylfarnesyl (fpp) cosubstrates of geranylgeranyl transferase and farnesyl transferase, and geranylgeranyl (ggti 287) and farnesyl phosphatase (fti 2 ... | 2011 | 21278325 |
| multiplex pcr assay for the detection of five putative virulence genes encoded in verotoxigenic escherichia coli plasmids. | the aim was to perform a pentavalent pcr assay for the detection of putative virulence genes encoded in vtec plasmids, katp, espp, suba, stce, and ehxa. the five-specific primer pairs used in the assay do not interfere with each other and generate amplification products of 914, 774, 556, 399, and 262 bp. it was selected at random 39 strains belonged to 20 serotypes in order to evaluate the multiplex in a wide variety of strains. the results of this study indicate that it is possible to perform s ... | 2011 | 21279513 |
| 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 ... | 2011 | 21279641 |
| inflammatory induction of human resistin causes insulin resistance in endotoxemic mice. | although adipocyte-derived murine resistin links insulin resistance to obesity, the role of human resistin, predominantly expressed in mononuclear cells and induced by inflammatory signals, remains unclear. given the mounting evidence that obesity and type 2 diabetes are inflammatory diseases, we sought to determine the relationship between inflammatory increases in human resistin and insulin resistance. | 2011 | 21282361 |
| [cloning and expression of a single human immunoglobulin heavy-chain variable domain with vascular endothelial growth factor binding activity]. | in the application of therapeutic antibodies, large molecular weight of antibodies is always a problem that prevents them from penetrating into tissues or binding to antigenic determinants. to overcome this problem, we investigated the function of the heavy chain variable domain of a monoclonal anti-vegf human igm antibody derived from the five-feature translocus mice. we cloned the cdna of the heavy chain variable domain, which was then inserted into pet28a vector and expressed in escherichia c ... | 2010 | 21284216 |
| [purification and characterization of recombinant human lactoferrin expressed in a cattle mammary bioreactor]. | novel ion exchange adsorbents were synthesized by immobilizing sulfopropyl derivative onto homemade highly cross-linked agarose beads. the effects of different ligand densities (from 0.05 to 0.24 mol/l) on static and dynamic adsorption of the adsorbents were investigated using lysozyme as a model protein. based on these results, rhlf was purified from the transgenic milk by our sp media. 1 ml high density (0.24 mol/l) adsorbent could handle 50 ml rhlf-containing milk. the mass recovery of rhlf w ... | 2010 | 21284219 |
| [production of the recombinant human bone morphogenetic protein-2 in escherichia coli and testing of its biological activity in vitro and in vivo]. | bone morphogenetic protein-2 (rhbmp-2) represents the osteoinductive protein factor which plays a dominant role in growth and regeneration of a bone tissue. in clinical practice the bone grafting materials on the basis of rhbmp-2 are widely applied; the russian analogues of similar materials are not produced. the fragment of the bmp2gene coding for a mature protein was cloned in escherichia coli. the effective overproducing strain of rhbmp-2 was created on a basis of the e. coli bl21 (de3). the ... | 2010 | 21290825 |
| human infections with non-o157 shiga toxin-producing escherichia coli, switzerland, 2000-2009. | we characterized 97 non-o157 shiga toxin (stx)-producing escherichia coli strains isolated from human patients during 2000-2009 from the national reference laboratory in switzerland. these strains belonged to 40 o:h serotypes; 4 serotypes (o26:h11/h-, o103:h2, o121:h19, and o145:h28/h-) accounted for 46.4% of the strains. nonbloody diarrhea was reported by 23.2% of the patients, bloody diarrhea by 56.8%. hemolytic uremic syndrome developed in 40.0% of patients; serotype o26:h11/h- was most often ... | 2011 | 21291586 |
| bone formation of block and particulated biphasic calcium phosphate lyophilized with escherichia coli-derived recombinant human bone morphogenetic protein 2 in rat calvarial defects. | the objective of this study was to evaluate bone formation in rat calvarial defects after surgical implantation of block or particulated biphasic calcium phosphate (bcp) lyophilized with escherichia coli-derived recombinant human bone morphogenetic protein 2 (erhbmp-2). critical-size calvarial osteotomy defects were created in 5 groups of sprague-dawley rats. each group received one of the following: 1) sham surgery control; 2) biphasic calcium phosphate particles (cpp); 3) biphasic calcium phos ... | 2011 | 21292513 |
| microbial evaluation and public health implications of urine as alternative therapy in clinical pediatric cases: health implication of urine therapy. | cultural means of pediatric treatment during ill health is a mainstay in africa, and though urine has been known to contain enteric pathogens, urine therapy is still culturally applicable in some health conditions and also advocated as alternative therapy. the study therefore, is to evaluate the microbial contents and safety of urine. | 2010 | 21293739 |
| enzymes of mannitol metabolism in the human pathogenic fungus aspergillus fumigatus--kinetic properties of mannitol-1-phosphate 5-dehydrogenase and mannitol 2-dehydrogenase, and their physiological implications. | the human pathogenic fungus aspergillus fumigatus accumulates large amounts of intracellular mannitol to enhance its resistance against defense strategies of the infected host. to explore their currently unknown roles in mannitol metabolism, we studied a. fumigatus mannitol-1-phosphate 5-dehydrogenase (afm1pdh) and mannitol 2-dehydrogenase (afm2dh), each recombinantly produced in escherichia coli, and performed a detailed steady-state kinetic characterization of the two enzymes at 25 °c and ph 7 ... | 2011 | 21299839 |
| molecular mimicry and ligand recognition in binding and catalysis by the histone demethylase lsd1-corest complex. | histone demethylases lsd1 and lsd2 (kdm1a/b) catalyze the oxidative demethylation of lys4 of histone h3. we used molecular dynamics simulations to probe the diffusion of the oxygen substrate. oxygen can reach the catalytic center independently from the presence of a bound histone peptide, implying that lsd1 can complete subsequent demethylation cycles without detaching from the nucleosomal particle. the simulations highlight the role of a strictly conserved active-site lys residue providing gene ... | 2011 | 21300290 |
| role of unique basic residues in cytotoxic, antibacterial and antiparasitic activities of human eosinophil cationic protein. | eosinophil granule proteins, eosinophil cationic protein (ecp) and eosinophil-derived neurotoxin are members of the rnase a superfamily, which play a crucial role in host defense against various pathogens as they are endowed with several biological activities. some of the biological activities possessed by ecp have been attributed to its strong basic character. in the current study, we have investigated the role of five unique basic residues, arg22, arg34, arg61, arg77 and his64 of ecp in its ca ... | 2011 | 21303303 |
| a secretory system for bacterial production of high-profile protein targets. | escherichia coli represents a robust, inexpensive expression host for the production of recombinant proteins. however, one major limitation is that certain protein classes do not express well in a biologically relevant form using standard expression approaches in the cytoplasm of e. coli. to improve the usefulness of the e. coli expression platform we have investigated combinations of promoters and selected n-terminal fusion tags for the extracellular expression of human target proteins. a compa ... | 2011 | 21308845 |
| ani s 10, a new anisakis simplex allergen: cloning and heterologous expression. | anisakiasis is a human disease caused by accidental ingestion of larval nematodes belonging to the anisakidae family. anisakiasis is often associated with a strong allergic response. diagnosis of a. simplex allergy is currently carried out by test based on the ige reactivity to a complete extract of l3 anisakis larvae although the specificity of these diagnostic tests is poor. improving the specificity of the diagnostic test is possible using purified recombinant allergens. a new anisakis allerg ... | 2011 | 21310242 |
| human papillomaviruses prevalence and genital co-infections in hiv-seropositive women in ouagadougou (burkina faso). | the vaginal swabs among hiv-positive women in africa often revealed opportunistic infections such as human papillomavirus (hpv) and mycoplasma that induce respectively cervix cancer and diseases such as vaginosis, abortions, infertility in through salpingitis. the purposes of this study were to: (1) seek for, the prevalence of pathogens such as hpv and mycoplasma; (2) characterize the strains of hpv and estimate their prevalence; (3) identify among these women, those who were co-infected by thes ... | 2010 | 21313918 |
| attenuation of human neutrophil migration and function by uropathogenic bacteria. | the establishment of bacterial infections at mucosal epithelial surfaces is determined by the balance of virulence attributes of the pathogen with the activity of innate host defenses. polymorphonuclear leukocytes (pmn) are key responders in many bacterial infections, but the mechanisms by which pathogens subvert these early responses to establish infection are largely undefined. here, we model early interactions between human pmn and the primary cause of urinary tract infections, namely uropath ... | 2011 | 21315174 |
| consequences of the overexpression of a eukaryotic membrane protein, the human kdel receptor, in escherichia coli. | escherichia coli is the most widely used host for producing membrane proteins. thus far, to study the consequences of membrane protein overexpression in e. coli, we have focussed on prokaryotic membrane proteins as overexpression targets. their overexpression results in the saturation of the sec translocon, which is a protein-conducting channel in the cytoplasmic membrane that mediates both protein translocation and insertion. saturation of the sec translocon leads to (i) protein misfolding/aggr ... | 2011 | 21316372 |
| expression, purification, and characterization of functional recombinant human daintain/aif-1 in escherichia coli. | daintain/aif-1 was identified from injured rat carotid arteries and porcine intestine in the mid 1990s. it is involved in autoimmune disorders, chronic rejection of allografts, gliomas, and breast cancer. since it is convenient and economical to obtain such a peptide biologically, in this study, we describe the expression, purification, and characterization of recombinant human daintain/aif-1 (rhdaintain/aif-1). the backbone of vector pet32a, a high-level expression plasmid, was used to construc ... | 2010 | 21319716 |
| a flagellin-producing lactococcus strain: interactions with mucin and enteropathogens. | bacillus cereus ch is a probiotic strain used in human nutrition whose adhesion to mucin is dependent on its surface-associated flagellin. flagellins from the surface of several probiotic bacillus strains were efficiently extracted with 5 m licl and identified by peptide fingerprinting. based on the proteomic analysis, cloning of the gene coding for the flagellin of b. cereus ch was performed in the lactococcal vector pnz8110 under the control of a nisin-inducible promoter. the resulting strain, ... | 2011 | 21323981 |
| integrated analysis of water quality parameters for cost-effective faecal pollution management in river catchments. | in many parts of the world, microbial contamination of surface waters used for drinking, recreation, and shellfishery remains a pervasive risk to human health, especially in less economically developed countries (ledc). however, the capacity to provide effective management strategies to break the waterborne route to human infection is often thwarted by our inability to identify the source of microbial contamination. microbial source tracking (mst) has potential to improve water quality managemen ... | 2011 | 21324505 |
| structure-activity relationship of 2,4,5-trioxoimidazolidines as inhibitors of thymidine phosphorylase. | novel non-nucleobase-derived inhibitors of the angiogenic enzyme, thymidine phosphorylase, have been identified using molecular modelling, synthesis and biological evaluation. these inhibitors are 2,4,5-trioxoimidazolidines bearing n-(substituted)phenylalkyl groups, together with, in most cases, n'-(ch(2))(n)-carboxylic acid, ester or amide side chains. the best compound from this series is 3-(2,4,5-trioxo-3-phenylethyl-imidazolodin-1-yl)propionamide, with an ic(50) of 40 µm against escherichia ... | 2011 | 21324566 |
| proinflammatory fecal mrna and childhood bacterial enteric infections. | introduction: assessment of specific mrnas in human samples is useful in characterizing disease. however, mrna in human stool has been understudied. results: compared to controls, infected stools showed increased transcripts of il-1β, il-8 and calprotectin. mrna and protein concentrations correlated for il-8, but not for calprotectin. discussion: stool mrna quantification offers a potentially useful, noninvasive way to assess inflammation in the gastrointestinal tract, and may be more sensitive ... | 2010 | 21327027 |
| evaluation of the antibacterial and anticancer activities of some south african medicinal plants. | several herbs are traditionally used in the treatment of a variety of ailments particularly in the rural areas of south africa where herbal medicine is mainly the source of health care system. many of these herbs have not been assessed for safety or toxicity to tissue or organs of the mammalian recipients. | 2011 | 21329526 |
| molecular cloning, characterization, and immunolocalization of two lactate dehydrogenase homologous genes from taenia solium. | two novel genes encoding lactate dehydrogenase a (ldha) and b (ldhb) homologues, respectively, were identified from the cdna libraries of adult taenia solium (t. solium). the two deduced amino acid sequences both show more than 50% identity to the homologues for danio rerio, xenopus laevis, schistosoma japonicum, sus scrofa, homo sapiens, et al. the identity of the amino acid sequence between tsldha and tsldhb is 57.4%, and that of the nucleotide sequence is 61.5%. recombinant tsldha homologue ( ... | 2011 | 21331787 |
| mammalian forebrain ketimine reductase identified as µ-crystallin; potential regulation by thyroid hormones. | ketimine reductase (e.c. 1.5.1.25) was purified to apparent homogeneity from lamb forebrain by means of a rapid multi-step chromatography protocol. the purified enzyme was identified by ms/ms (mass spectrometry) as µ-crystallin. the identity was confirmed by heterologously expressing human µ-crystallin in escherichia coli and subsequent chromatographic purification of the protein. the purified human µ-crystallin was confirmed to have ketimine reductase activity with a maximum specific activity s ... | 2011 | 21332720 |
| screening for antimicrobial resistance genes and virulence factors via genome sequencing. | second-generation genome sequencing and alignment of the resulting reads to in silico genomes containing antimicrobial resistance and virulence factor genes were used to screen for undesirable genes in 28 strains which could be used in human nutrition. no virulence factor genes were detected, while several isolates contained antimicrobial resistance genes. | 2011 | 21335393 |
| human serum albumin improves endothelial dysfunction and survival during experimental endotoxemia: concentration-dependent properties. | the mechanisms by which human serum albumin might protect against sepsis-induced organ dysfunction and improve survival are not elucidated. the present study was designed to assess the effects of two concentrations of human serum albumin on endotoxin-induced mortality as well as on endothelial and organ dysfunctions in both mouse and cell models. | 2011 | 21336119 |
| engineering of escherichia coli for targeted delivery of transgenes to her2/neu-positive tumor cells. | targeting of non-phagocytic tumor cells and prompt release of gene cargos upon entry into tumors are two limiting steps in the bacterial gene delivery path. to tackle these problems, the non-pathogenic escherichia coli strain bl21(de3) was engineered to display the anti-her2/neu affibody on the surface. after co-incubation with tumor cells for 3 h, the anti-her2/neu affibody-presenting e. coli strain was selectively internalized into her2/neu-positive skbr-3 cells. the invasion efficiency reache ... | 2011 | 21337329 |
| efficient production of soluble recombinant single chain fv fragments by a pseudomonas putida strain kt2440 cell factory. | recombinant antibody fragments have a wide range of applications in research, diagnostics and therapy. for many of these, small fragments like single chain fragment variables (scfv) function well and can be produced inexpensively in bacterial expression systems. although escherichia coli k-12 production systems are convenient, yields of different fragments, even those produced from codon-optimized expression systems, vary significantly. where yields are inadequate, alternative production systems ... | 2011 | 21338491 |
| biotinylation, a post-translational modification controlled by the rate of protein-protein association. | biotin protein ligases catalyze specific covalent linkage of the coenzyme biotin to biotin-dependent carboxylases. the reaction proceeds in two steps, including synthesis of an adenylated intermediate followed by biotin transfer to the carboxylase substrate. in this work specificity in the transfer reaction was investigated using single turnover stopped-flow and quench-flow assays. cognate and noncognate reactions were measured using the enzymes and minimal biotin acceptor substrates from escher ... | 2011 | 21343300 |
| phenotypic and transcriptional analysis of the osmotic regulator ompr in yersinia pestis. | the osmotic regulator ompr in escherichia coli regulates differentially the expression of major porin proteins ompf and ompc. in yersinia enterocolitica and y. pseudotuberculosis, ompr is required for both virulence and survival within macrophages. however, the phenotypic and regulatory roles of ompr in y. pestis are not yet fully understood. | 2011 | 21345178 |
| enterohaemorrhagic escherichia coli haemolysin is cleaved and inactivated by serine protease esppa. | the haemolysin from enterohaemorrhagic escherichia coli (ehec-hly) and the serine protease esppa are putative virulence factors of ehec. we investigated the interplay between these secreted factors and demonstrate that esppa cleaves the 107 kda large ehec-hly. degradation was observed when purified esppa was added to a growing culture of an ehec-hly-expressing strain, with isolated proteins and with coexpressing strains, and was independent of the ehec serotype. ehec-hly breakdown occurred as a ... | 2011 | 21352460 |
| architecture of the helicobacter pylori cag-type iv secretion system. | type iv secretion systems (t4ss) are macromolecular assemblies used by bacteria to transport material across their membranes. t4ss are generally composed of a set of twelve proteins (virb1-11 and vird4). this represents a dynamic machine powered by three atpases. t4ss are widespread in pathogenic bacteria where they are often used to deliver effectors into host cells. for example, the human pathogen helicobacter pylori encodes a t4ss, the cag-t4ss, which mediates the injection of the toxin caga. ... | 2011 | 21352491 |
| broad-spectrum antimicrobial efficacy of peptide a3-apo in mouse models of multidrug-resistant wound and lung infections cannot be explained by in vitro activity against the pathogens involved. | although the designer proline-rich antimicrobial peptide a3-apo has only modest activity against escherichia coli and acinetobacter baumannii in vitro, in mouse models of systemic and wound infections it shows superior efficacy compared with conventional antibiotics. in this study, the efficacy of a3-apo in several additional mouse models was investigated, including staphylococcus aureus wound infection, mixed klebsiella pneumoniae-a. baumannii-proteus mirabilis wound infection and k. pneumoniae ... | 2011 | 21353493 |
| a novel single-chain variable fragment antibody against fgf-1 inhibits the growth of breast carcinoma cells by blocking the intracrine pathway of fgf-1. | the fibroblast growth factors (fgfs) are important for embryo development, wound healing, hematopoiesis, and angiogenesis. fgf-1, a member of fgf family, is involved in both receptor-dependent pathways and an intracrine pathway. studies have recently shown that fgf-1 is overexpressed in the early stages of several kinds of cancer. thus, fgf-1 is a candidate for cancer immunotargeting. to study the potential use of therapeutic antibodies against fgf-1, a monoclonal hybridoma 1c9 secreting monoclo ... | 2011 | 21360642 |
| strategy for microbiome analysis using 16s rrna gene sequence analysis on the illumina sequencing platform. | understanding the identity and changes of organisms in the urogenital and other microbiomes of the human body may be key to discovering causes and new treatments of many ailments, such as vaginosis. high-throughput sequencing technologies have recently enabled discovery of the great diversity of the human microbiome. the cost per base of many of these sequencing platforms remains high (thousands of dollars per sample); however, the illumina genome analyzer (iga) is estimated to have a cost per b ... | 2011 | 21361774 |
| cyanobacterial microcystis aeruginosa lipopolysaccharide elicits release of superoxide anion, thromboxane b2, cytokines, chemokines, and matrix metalloproteinase-9 by rat microglia. | microcystis aeruginosa (m. aeruginosa) is a cosmopolitan gram-negative cyanobacterium that may contaminate freshwater by releasing toxins, such as lipopolysaccharide (lps) during aquatic blooms, affecting environmental and human health. the putative toxic effects of cyanobacterial lps on brain microglia, a glial cell type that constitutes the main leukocyte-dependent source of reactive oxygen species in the central nervous system, are presently unknown. we tested the hypothesis that in vitro con ... | 2011 | 21362633 |
| bacterial conversion of folinic acid is required for antifolate resistance. | antifolates, which are among the first antimicrobial agents invented, inhibit cell growth by creating an intracellular state of folate deficiency. clinical resistance to antifolates has been mainly attributed to mutations that alter structure or expression of enzymes involved in de novo folate synthesis. we identified a mycobacterium smegmatis mutant, named fuel (which stands for folate utilization enzyme for leucovorin), that is hypersusceptible to antifolates. chemical complementation indicate ... | 2011 | 21372133 |
| analogues of peptide smap-29 with comparable antimicrobial potency and reduced cytotoxicity. | smap-29 (sheep myeloid antimicrobial peptide-29) is a peptide with potent antibacterial properties. however, it is also highly cytotoxic both to human red blood cells (hrbcs) and human embryonic kidney (hek) cells. in this study, some of the amino acids of smap-29 were changed in an attempt to reduce haemolytic activity whilst maintaining high antibacterial efficacy. these analogues, plus other analogues described in the literature with potent antimicrobial activity against gram-positive bacteri ... | 2011 | 21377841 |
| e. coli nissle 1917 affects salmonella adhesion to porcine intestinal epithelial cells. | the probiotic escherichia coli strain nissle 1917 (ecn) has been shown to interfere in a human in vitro model with the invasion of several bacterial pathogens into epithelial cells, but the underlying molecular mechanisms are not known. | 2011 | 21379575 |
| baking a mass-spectrometry data pie with mcmc and simulated annealing: predicting protein post-translational modifications from integrated top-down and bottom-up data. | post-translational modifications are vital to the function of proteins, but are hard to study, especially since several modified isoforms of a protein may be present simultaneously. mass spectrometers are a great tool for investigating modified proteins, but the data they provide is often incomplete, ambiguous and difficult to interpret. combining data from multiple experimental techniques-especially bottom-up and top-down mass spectrometry-provides complementary information. when integrated wit ... | 2011 | 21389073 |
| identification and conformer analysis of a novel redox-active motif, pro-ala-ser-cys-cys-ser, in drosophila thioredoxin reductase by semiempirical molecular orbital calculation. | mammalian thioredoxin reductases (trxrs) contain selenium as selenocysteine (sec) in the c-terminal redox center -gly-cys-sec-gly-oh to reduce trx and other substrates; a sec-to-cys substitution in mammalian trxr yields an almost inactive enzyme. the corresponding tetrapeptide sequence in drosophila melanogaster trxr (dm-trxr), -ser-cys-cys-ser-oh, endows the orthologous enzyme with a catalytic competence similar to mammalian selenoenzymes, but implementation of the ser-containing tetrapeptide s ... | 2011 | 21389620 |
| influence of 2% chlorhexidine gel on calcium hydroxide ionic dissociation and its ability of reducing endotoxin. | the aim of this study was to evaluate the influence of 2% chlorhexidine gel (chx) on ph, calcium release, and ca(oh)(2) capability of reducing endotoxin. | 2011 | 21393032 |
| muc17 protects intestinal epithelial cells from enteroinvasive e. coli infection by promoting epithelial barrier integrity. | the membrane-bound mucin muc17 (mouse homolog muc3) is highly expressed on the apical surface of intestinal epithelia and is thought to play a role in epithelial restitution and protection. therefore, we hypothesized that muc17 has a role in protection of the intestinal mucosa against luminal pathogens. human intestinal cell lines were transfected by electroporation (caco-2 and ht 29/19a) and by retroviral expression vector (ls174t, a cell line with high levels of muc17 expression) using muc17 s ... | 2011 | 21393431 |
| passive vaccination with a human monoclonal antibody: generation of antibodies and studies for efficacy in bacillus anthracis infections. | a major difficulty in creating human monoclonal antibodies is the lack of a suitable myeloma cell line to be used for fusion experiments. in order to create fully human monoclonal antibodies for passive immunization, the human mouse heteromyeloma cell line cb-f7 was evaluated. using this cell line, we generated human monoclonal antibodies against bacillus anthracis toxin components. antibodies against protective antigen (pa) and against lethal factor (lf) were obtained using peripheral blood lym ... | 2010 | 21397977 |
| ionic liquid promoted synthesis, antibacterial and in vitro antiproliferative activity of novel +¦-aminophosphonate derivatives. | ionic liquid ethyl ammonium nitrate is used as an excellent catalyst and solvent for three-component one-pot reaction of an aldehydes, amines and diethylphosphite to form novel +¦-aminophosphonates at room temperature. among the various catalysts, the preparation of ethyl ammonium nitrate is an environmental friendly, cost effective and recyclable catalyst. compounds 4b, 4c, 4d, 4f and 4j were found more potent antibacterials against pathogenic microorganisms. whereas, compounds 4a, 4g, 4h and 4 ... | 2011 | 21398120 |
| prevalence of faecal esbl carriage in the community and in a hospital setting in a county of southern sweden. | the aim of this study was to investigate the prevalence of extended-spectrum beta-lactamase (esbl)-producing bacteria in patients at various hospital wards and in a group of relatively healthy volunteers, in order to obtain greater knowledge on how common these bacterial strains are in hospital settings and in the general community. participants (nôçë=ôçë427) were enrolled at a university hospital and at primary health care units (phcus) in sweden in 2008 and 2010. the participants provided rect ... | 2011 | 21399889 |
| differences in toll-like receptor expression and cytokine production after stimulation with heat-killed gram-positive and gram-negative bacteria. | innate immune surveillance in the blood is executed mostly by circulating monocytes, which recognize conserved bacterial molecules such as peptidoglycan and lipopolysaccharide. toll-like receptors (tlr) play a central role in microbe-associated molecular pattern detection. the aim of this study was to compare the differences in tlr expression and cytokine production after stimulation of peripheral blood cells with heat-killed gram-negative and gram-positive human pathogens: neisseria meningitidi ... | 2011 | 21399944 |
| trimerization of murine tnf ligand family member light increases the cytotoxic activity against the fm3a mammary carcinoma cell line. | light is a member of the tumor necrosis factor ligand superfamily, which plays important roles in inflammatory and immune responses. light forms a membrane-anchored homotrimeric complex on the cell surface and is often processed as a soluble protein. recombinant soluble human light produced by mammalian cells or escherichia coli is functional at nanomolar concentrations. however, there is little information about the biological activity of mouse light (mlight) because of the difficulty in produc ... | 2011 | 21400099 |
| liposomes containing glucosyl ceramide specifically bind t4 bacteriophage: a self-assembling nanocarrier formulation. | a unique formulation is described comprising liposomes containing glucosyl ceramide (glucer) in the lipid bilayer to which bacteriophage t4 was attached. binding of the phage t4 did not occur to glycolipids, such as galactosyl ceramide, containing an aldose in which the c-2 or c-4 conformations were not identical to glucose. these results strongly support previous proposals that glucose is a major receptor moiety for t4 binding to escherichia coli. by using the binding of t4 to liposomal glucer, ... | 2011 | 21401373 |
| recombinant heteromeric phenylalanine monooxygenase and the oxygenation of carbon and sulfur substrates. | the aim of this investigation was to provide in-vitro enzyme kinetic data to support the hypothesis that the in-vivo heterozygous dominant phenotype for phenylalanine monooxygenase (hpah) was responsible for the s-oxidation polymorphism in the metabolism of s-carboxymethyl-l-cysteine reported in humans. using a dual-vector expression strategy for the co-production of wild-type and mutant human hpah subunits we report for the first time the kinetic parameters (k(m) , v(max) , cl(e) ) for the c-ox ... | 2011 | 21401608 |
| what makes a natural clay antibacterial? | natural clays have been used in ancient and modern medicine, but the mechanism(s) that make certain clays lethal against bacterial pathogens has not been identified. we have compared the depositional environments, mineralogies, and chemistries of clays that exhibit antibacterial effects on a broad spectrum of human pathogens including antibiotic resistant strains. natural antibacterial clays contain nanoscale (<200 nm), illite-smectite and reduced iron phases. the role of clay minerals in the ba ... | 2011 | 21413758 |
| the molecular basis of aminoacylase 1 deficiency. | aminoacylase 1 is a zinc-binding enzyme which hydrolyzes n-acetyl amino acids into the free amino acid and acetic acid. deficiency of aminoacylase 1 due to mutations in the aminoacylase 1 (acy1) gene follows an autosomal-recessive trait of inheritance and is characterized by accumulation of n-acetyl amino acids in the urine. in affected individuals neurological findings such as febrile seizures, delay of psychomotor development and moderate mental retardation have been reported. except for one m ... | 2011 | 21414403 |
| ntp technical report on the toxicology and carcinogenesis studies of beta-myrcene (cas no. 123-35-3) in f344/n rats and b6c3f1 mice (gavage studies). | beta-myrcene, an acyclic unsubstituted monoterpene, and the essential oils which contain it are used as intermediates in the production of terpene alcohols (geraniol, nerol, and linalool), which, in turn, serve as intermediates in the production of aroma and flavor chemicals. thus beta-myrcene is used widely in cosmetics, soaps, and detergents and as a flavoring additive in food and beverages. beta-myrcene is also the major constituent of hop and bay oils, which are used in the manufacture of al ... | 2010 | 21415873 |
| effects of enrofloxacin on the human intestinal microbiota in vitro. | an anaerobic, continuous-flow culture method has been developed to assess the effects of different levels of enrofloxacin (enr) on the human intestinal microbiota. chemostats containing human faecal flora were exposed to 1.25, 12.5 and 125 µg/ml enr. before and during drug exposure, samples aspirated from culture vessels were analysed using viable cell counting and denaturing gradient gel electrophoresis (dgge). in addition, the colonisation resistance (cr) of the microbiota to candida albicans ... | 2011 | 21420834 |
| production and solid-phase refolding of human glucagon-like peptide-1 using recombinant escherichia coli. | human glucagon-like peptide-1 (glp-1), an incretin hormone with pharmaceutical potential in treating type 2 diabetes mellitus is known to be rapidly degraded when expressed in escherichia coli. for the efficient production of the intact glp-1 using a recombinant e. coli system, a fusion protein of glp-1 was designed to be composed of the 6-lysine tag, ubiquitin and glp-1 (k6ubglp-1) in a row. a fed-batch fermentation of recombinant e. coli bl21(de3)/papk6ubglp-1 was carried out in a ph-stat feed ... | 2011 | 21421051 |
| substituted imidazole of 5-fluoro-2-[4-[(2-phenyl-1h-imidazol-5-yl)methyl]-1-piperazinyl]pyrimidine inactivates cytochrome p450 2d6 by protein adduction. | 5-fluoro-2-[4-[(2-phenyl-1h-imidazol-5-yl)methyl]-1-piperazinyl]pyrimidine (sch 66712) is a potent mechanism-based inactivator of human cytochrome p450 2d6 that displays type i binding spectra with a k(s) of 0.39 ± 0.10 µm. the partition ratio is ~3, indicating potent inactivation that addition of exogenous nucleophiles does not prevent. within 15 min of incubation with sch 66712 and nadph, ~90% of cyp2d6 activity is lost with only ~20% loss in ability to bind co and ~25% loss of native heme ove ... | 2011 | 21422192 |
| mo25 is a master regulator of spak/osr1 and mst3/mst4/ysk1 protein kinases. | mouse protein-25 (mo25) isoforms bind to the strad pseudokinase and stabilise it in a conformation that can activate the lkb1 tumour suppressor kinase. we demonstrate that by binding to several ste20 family kinases, mo25 has roles beyond controlling lkb1. these new mo25 targets are spak/osr1 kinases, regulators of ion homeostasis and blood pressure, and mst3/mst4/ysk1, involved in controlling development and morphogenesis. our analyses suggest that mo25a and mo25ß associate with these ste20 kina ... | 2011 | 21423148 |
| monitoring of multiple drug-resistant pathogens in a selected stretch of bay of bengal, india. | the present work aims at identification of multiple drug-resistant pathogenic bacteria in a selected stretch, namely, puri on the bay of bengal, india. six stations at the coast of puri were selected and samples of water and sediment were collected during the winter of 2008 and 2009 for this study. thirty-eight pathogenic bacteria were isolated and identified from both the water and the sediment of 6 fixed stations (pu-1a, pu-1b, pu-2, pu-3, pu-4, and pu-5). the identified pathogens were escheri ... | 2011 | 21424669 |
| [a man with unusual abdominal gas]. | a 34 year old male was admitted to our hospital with a severe pancreatitis. on a ct scan we diagnosed a developing emphysematous pancreatitis. this is a rare form of pancreatitis usually caused by an infection with an escherichia coli. | 2010 | 21429258 |
| identification and safety evaluation of bacillus species occurring in high numbers during spontaneous fermentations to produce gergoush, a traditional sudanese bread snack. | gergoush is a naturally fermented sudanese bread snack produced in three fermentation steps (primary starter, adapted starter and final dough), followed by three baking steps for a half to one hour at above 200 °c. this study examines the microbiota of two sets of fermentations performed at a traditional production site in khartoum, sudan in 2006 and 2009, respectively. in 2006 four different milk/legume based primary starters (faba bean, chick pea, lentil and white bean) were sampled in order t ... | 2011 | 21429611 |
| expression of human c-reactive protein in different systems and its purification from leishmania tarentolae. | with its homo-pentameric structure and calcium-dependent specificity for phosphocholine (pch), human c-reactive protein (crp) is produced by the liver and secreted in elevated quantities in response to inflammation. crp is widely accepted as a cardiac marker, e.g. in point-of-care diagnostics, however, its heterologous expression has proven difficult. here, we demonstrate the expression of crp in different escherichia coli strains as well as by in vitro transcription/translation. although expres ... | 2011 | 21440634 |
| high-yield resveratrol production in engineered escherichia coli. | plant polyphenols have been the subject of several recent scientific investigations since many of the molecules in this class have been found to be highly active in the human body, with a plethora of health-promoting activities against a variety of diseases, including heart disease, diabetes, and cancer, and with even the potential to slow aging. further development of these potent natural therapeutics hinges on the formation of robust industrial production platforms designed using specifically ... | 2011 | 21441338 |
| chlamydia trachomatis secretion of an immunodominant hypothetical protein (ct795) into host cell cytoplasm. | the chlamydia-specific hypothetical protein ct795 was dominantly recognized by human antisera produced during c. trachomatis infection but not by animal antisera raised against dead chlamydia organisms. the immundominant region recognized by the human antibodies was mapped to the n-terminal fragment t22-s69. the endogenous ct795 was detected in the cytoplasm of host cells during c. trachomatis infection and was highly enriched in the host cytosolic fraction but absent in the purified chlamydia o ... | 2011 | 21441519 |
| nucleosomal h2b ubiquitylation with purified factors. | diverse histone modifications play important roles in transcriptional regulation throughout eukaryotes, and recent studies have implicated histone h2b ubiquitylation in active transcription. the necessity of at least three enzymes (e1-e3), as well as ongoing transcription events, for efficient h2b ubiquitylation complicates mechanistic studies of h2b ubiquitylation relative to other histone modifications. here we describe experimental protocols for preparation of human h2b ubiquitylation factors ... | 2011 | 21443952 |
| complex principal component and correlation structure of 16 yeast genomic variables. | a quickly growing number of characteristics reflecting various aspects of gene function and evolution can be either measured experimentally or computed from dna and protein sequences. the study of pairwise correlations between such quantitative genomic variables as well as collective analysis of their interrelations by multidimensional methods have delivered crucial insights into the processes of molecular evolution. here, we present a principal component analysis (pca) of 16 genomic variables f ... | 2011 | 21444651 |
| expression of human globular adiponectin-glucagon-like peptide-1 analog fusion protein and its assay of glucose-lowering effect in vivo. | in this study, human globular adiponectin-glucagon-like peptide-1 analog (gad-glp-1-a) fusion protein was expressed and its glucose-lowering effect was measured in vivo. we constructed a prokaryotic expression vector pet28a-gad-glp-1-a and transformed the vector into escherichia coli bl21 (de3). a recombinant fusion protein of about 25kd was expressed from bl21 (de3) cells after isopropylthio-ß-d-galactoside induction. this protein was n-terminal his-tagged gad-glp-1-a fusion protein. most of th ... | 2011 | 21448306 |
| 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 ... | 2011 | 21449573 |
| solution nmr insights into docking interactions involving inactive erk2. | the mitogen-activated protein (map) kinase erk2 contains recruitment sites that engage canonical and noncanonical motifs found in a variety of upstream kinases, regulating phosphatases and downstream targets. interactions involving two of these sites, the d-recruitment site (drs) and the f-recruitment site (frs), have been shown to play a key role in signal transduction by erk/map kinases. the dynamic nature of these recruitment events makes nmr uniquely suited to provide significant insight int ... | 2011 | 21449613 |
| interplay between the acute inflammatory response and heart rate variability in healthy human volunteers. | the autonomic nervous system and the inflammatory response are intimately linked. heart rate variability (hrv) analysis is a widely used method to assess cardiac autonomic nervous system activity, and changes in hrv indices may correlate with inflammatory markers. here, we investigated whether baseline hrv predicts the acute inflammatory response to endotoxin. second, we investigated whether the magnitude of the inflammatory response correlated with hrv alterations. forty healthy volunteers rece ... | 2011 | 21451444 |
| inactivation of escherichia coli o157:h7 attached to spinach harvester blade using bacteriophage. | outbreaks associated with leafy greens have focused attention on the transfer of human pathogens to these commodities during harvest with commercial equipment. attachment of escherichia coli o157:h7 on new or rusty spinach harvester blades immersed in spinach extract or 10% tryptic soy broth (tsb) was investigated. bacteriophages specific for e. coli o157:h7 were evaluated to kill cells attached to blade. a cocktail of five nalidixic acid-resistant e. coli o157:h7 isolates was transferred to 25 ... | 2011 | 21453119 |
| regulatory circuits of the aaa+ disaggregase hsp104. | yeast hsp104 is an aaa+ chaperone that rescues proteins from the aggregated state. six protomers associate to form the functional hexamer. each protomer contains two aaa+ modules, nbd1 and nbd2. hsp104 converts energy provided by atp into mechanical force used to thread polypeptides through its axial channel, thereby disrupting protein aggregates. but how the action of its 12 aaa+ domains is co-ordinated to catalyze disaggregation remained unexplained. here, we identify a sophisticated allosteri ... | 2011 | 21454552 |
| identification of constitutively active interleukin 33 (il-33) splice variant. | il-33/il-1f11 is a new member of the il-1 family ligand and provokes t helper-type immune responses. il-33 is the ligand of st2 and il-1 receptor accessory protein (il-1racp) that triggers nuclear factor-? light chain enhancer of activated b cells (nf-?b) and mapk signaling. we discovered a novel short splice variant of il-33 that was termed spil-33. the new spil-33 lacks exon 3 containing a proposed caspase-1 cleavage site. we isolated spil-33 cdna from the huh7 human hepatocarcinoma cell line ... | 2011 | 21454686 |
| crystal structure of the central coiled-coil domain from human liprin-β2. | liprins are a conserved family of scaffolding proteins important for the proper regulation and development of neuronal synapses. humans have four liprin-αs and two liprin-βs which all contain long coiled-coil domains followed by three tandem sam domains. complex interactions between the coiled-coil and sam domains are thought to create liprin scaffolds, but the structural and biochemical properties of these domains remain largely uncharacterized. in this study we find that the human liprin-β2 co ... | 2011 | 21462929 |
| characterization of the e506q and h537a dysfunctional mutants in the e. coli abc transporter msba. | msba is a member of the abc transporter superfamily that is specifically found in gram-negative bacteria and is homologous to proteins involved in both bacterial and human drug resistance. the e506q and h537a mutations have been introduced and used for crystallization of other members of the abc transporter protein family, including bmra and the atpase domains malk, hlyb-nbd, and mj0796, but have not been previously studied in detail or investigated in the msba lipid a exporter. we utilized an a ... | 2011 | 21462989 |
| dutch patients, retail chicken meat and poultry share the same esbl genes, plasmids and strains. | intestinal carriage of extended-spectrum beta-lactamase (esbl) -producing bacteria in food-producing animals and contamination of retail meat may contribute to increased incidences of infections with esbl-producing bacteria in humans. therefore, distribution of esbl genes, plasmids and strain genotypes in escherichia coli obtained from poultry and retail chicken meat in the netherlands was determined and defined as 'poultry-associated' (pa). subsequently, the proportion of e. coli isolates with ... | 2011 | 21463397 |
| real-time attack on single escherichia coli cells by the human antimicrobial peptide ll-37. | natural antimicrobial peptides (amps) provide prototypes for the design of unconventional antimicrobial agents. existing bulk assays measure amp activity but do not provide details of the growth-halting mechanism. we use fluorescence microscopy to directly observe the attack of the human antimicrobial peptide ll-37 on single escherichia coli cells in real time. our findings strongly suggest that disruption of the cytoplasmic membrane is not the growth-halting mechanism. at 8 µm, ll-37 binding sa ... | 2011 | 21464330 |
| preparation and preliminary characterization of rabbit monoclonal antibodies against human midkine. | we prepared rabbit monoclonal antibodies that target human midkine (mk). the mk gene was amplified by pcr from the plasmid pegfp-mk and subcloned into the prokaryotic expression vector pgex-1?t to generate an n-terminally glutathione s-transferase (gst)-tagged fusion protein construct. expression of the gst-mk fusion protein was achieved by iptg induction in escherichia coli cells. the expressed protein was purified using the gst system. after verifying purification, the fusion protein was used ... | 2011 | 21466290 |
| nmr relaxation unravels interdomain crosstalk of the two domain prolyl isomerase and chaperone slyd. | the dynamics of the two domain prolyl-peptidyl cis/trans isomerase and chaperone slyd was studied on a ps-to-ns time scale to correlate dynamic changes with the catalytic function. (15)n transversal and longitudinal relaxation rates as well as heteronuclear overhauser effects were determined at different temperatures for escherichia coli slyd (ecslyd) and for thermus thermophilus slyd (ttslyd). with the well established extended lipari-szabo approach, the order parameter, s(2), the internal corr ... | 2011 | 21466862 |
| high-throughput analysis of the mutagenic and cytotoxic properties of dna lesions by next-generation sequencing. | human cells are constantly exposed to environmental and endogenous agents which can induce damage to dna. understanding the implications of these dna modifications in the etiology of human diseases requires the examination about how these dna lesions block dna replication and induce mutations in cells. all previously reported shuttle vector-based methods for investigating the cytotoxic and mutagenic properties of dna lesions in cells have low-throughput, where plasmids containing individual lesi ... | 2011 | 21470959 |
| a genetically encoded probe for the identification of proteins that form sulfenic acid in response to h(2)o(2) in saccharomyces cerevisiae. | it is widely known that reactive oxygen species (ros), such as hydrogen peroxide, play important roles in cellular signaling and initiation of oxidative stress responses via thiol modifications. identification of the targets of these modifications will provide a better understanding of the relationship between ros and human diseases, such as cancer and atherosclerosis. sulfenic acid is the principle product of a reaction between hydrogen peroxide and a reactive protein cysteine. this reversible ... | 2011 | 21476607 |
| statistical analysis of features associated with protein expression/solubility in an in vivo escherichia coli expression system and a wheat germ cell-free expression system. | recombinant protein technology is an important tool in many industrial and pharmacological applications. although the success rate of obtaining soluble proteins is relatively low, knowledge of protein expression/solubility under "standard" conditions may increase the efficiency and reduce the cost of proteomics studies. in this study, we conducted a genome-scale experiment to assess the overexpression and the solubility of human full-length cdna in an in vivo escherichia coli expression system a ... | 2011 | 21478485 |
| genome sequencing of environmental escherichia coli expands understanding of the ecology and speciation of the model bacterial species. | defining bacterial species remains a challenging problem even for the model bacterium escherichia coli and has major practical consequences for reliable diagnosis of infectious disease agents and regulations for transport and possession of organisms of economic importance. e. coli traditionally is thought to live within the gastrointestinal tract of humans and other warm-blooded animals and not to survive for extended periods outside its host; this understanding is the basis for its widespread u ... | 2011 | 21482770 |