Publications

apolipoprotein e is highly susceptible to oxidation by myeloperoxidase, an enzyme present in the brain.	apolipoprotein e, the most common apolipoprotein found in the brain, is linked to several pathologies like alzheimer's disease. apolipoprotein e directly binds to beta-amyloid with a strong affinity. myeloperoxidase, a protein secreted by neutrophils and involved in the inflammatory process, is also present in the brain. in vitro myeloperoxidase oxidation of recombinant human apolipoprotein e leads to fragmentation of the protein with low concentrations of hydrogen peroxide and polymerization wi ...	1996	8762192
isolation and characterization of recombinant human apolipoprotein c-ii expressed in escherichia coli.	a full-length recombinant human apolipoprotein c-ii (apoc-ii) has been successfully expressed in escherichia coli using the t7 expression system. the recombinant apoc-ii. which was expressed intracellularly in the inclusion bodies, was solubilized with 8 m urea and purified using sephadex g-75 gel permeation chromatography. four liters of the bacterial culture yielded 16-20 mg of purified recombinant apoc-ii. sequencing and mass spectrometric analyses indicated that the isolated recombinant apoc ...	1996	8765143
intrauterine bacterial findings and hormonal profiles in post-partum cows with normal puerperium.	the post-partum intrauterine bacterial flora, prostaglandin release, uterine involution and resumption of ovarian activity were studied in 9 swedish dairy cows during the first 8-week period. uterine involution was monitored by transrectal examinations of the reproductive tract 3 times weekly. bacteriological examination was performed from twice weekly uterine biopsies. the main pgf2 alpha metabolite (15-ketodihydro-pgf2 alpha) was monitored from twice daily blood plasma samples, while morning s ...	1996	8996871
[utilization of nylon membranes for specific isolation and characterization of verotoxin-producing escherichia coli using dna probes].	a method for specific isolation of vt(+)-strains in raw milk is given. dna-hybridization technique with dig-labeled pcr-amplificates as probes are the basis. no background is seen by using "dig easy hyb" solution and nylon membranes for colony- and plaque-hybridization (boehringer mannheim gmbh). marked colonies are visible on the membranes after detection. so it is possible to select these colonies from a masterplate. the results are available within one day (without enrichment and membrane pre ...	1996	8999776
expression of the colh gene encoding clostridium histolyticum collagenase in bacillus subtilis and its application to enzyme purification.	the colh gene encoding 116-kda collagenase of clostridium histolyticum (ccolh) was cloned into an escherichia coli-bacillus subtilis shuttle vector to develop a method for purification of recombinant collagenase (rcolh). when plasmid pjcm310 containing the colh gene was introduced into b. subtilis db104 and the transformant was grown in lb broth at 37 c, stability of the plasmid was not maintained. however, stability was partly improved by growing the transformant in a modified lb broth containi ...	1996	9013490
escherichia coli expression and substrate specificities of canine cytochrome p450 3a12 and rabbit cytochrome p450 3a6.	high level escherichia coli expression of cytochromes p450 3a12 and 3a6 has facilitated the characterization of proteins which exhibit limited activity as purified hepatic enzymes in reconstituted systems. three 3a12 and two 3a6 constructs modified at the 5'-end to encode the bovine 17 alpha-sequence (barnes et al., proc. natl. acad. sci. u.s.a. 88: 5597-5601, 1991), or related sequences, exhibited expression levels ranging from 2 to 89 nmol of cytochrome p450 liter-1. recombinant canine 3a12 ca ...	1996	8768753
downregulation of endothelial constitutive nitric oxide synthase expression by lipopolysaccharide.	lipopolysaccharide (lps), a causal agent of sepsis, has been shown to induce systemic nitric oxide (no) synthesis through complex mechanisms. however, the effect of lps on endothelial cells is incompletely understood. to investigate the mechanism by which lps influences the release of no from endothelial cells, the effect of this compound on endothelial constitutive nitric oxide synthase (ecnos) was studied in cultured bovine coronary venular endothelial cells. western and northern analyses show ...	1996	8769085
fate of enterohemorrhagic escherichia coli o157:h7 in bovine feces.	dairy cattle have been identified as a principal reservoir of escherichia coli o157:h7. the fate of this pathogen in bovine feces at 5, 22, and 37 degrees c was determined. two levels of inocula (10(3) and 10(5) cfu/g) of a mixture of five nalidixic acid-resistant e. coli o157:h7 strains were used. e. coli o157:h7 survived at 37 degrees c for 42 and 49 days with low and high inocula, respectively, and at 22 degrees c for 49 and 56 days with low and high inocula, respectively. fecal samples at bo ...	1996	8779595
epidemiology & management of persistent diarrhoea in children of developing countries.	diarrhoea that begins acutely but lasts longer than two weeks is defined to be persistent. revised estimates in developing countries including india showed that acute diarrhoea accounts for 35 per cent, dysentery 20 per cent and non-dysenteric persistent diarrhoea (pd) for 45 per cent of total diarrhoeal deaths. pd also often changes marginal malnutrition to more severe forms. factors that increase the risk of acute diarrhoea becoming persistent have been identified in india and other developing ...	1996	8783512
alanine point-mutations in the reactive region of bovine pancreatic trypsin inhibitor: effects on the kinetics and thermodynamics of binding to beta-trypsin and alpha-chymotrypsin.	in an effort to relate structural, kinetic, and thermodynamic features in a model macromolecular recognition process, the amino acid residues in the reactive surface of bovine pancreatic trypsin inhibitor (bpti) and surrounding residues were substituted individually by alanine, and the effects of the point-mutations on the kinetics and thermodynamics of inhibition by bpti toward trypsin and chymotrypsin were investigated. fifteen alanine mutants were produced. the majority of the bpti mutants ex ...	1996	8784199
escherichia coli o157:h7 in microbial flora of sheep.	we found naturally occurring, potentially virulent escherichia coli o157:h7 strains in sheep. the incidence of e. coli o157:h7 was transient and ranged from 31% of sheep in june to none in november. the use of a sensitive culture technique and the choice of the proper sampling season were both essential for detecting this bacterium in sheep. dna hybridizations showed that 80% of the e. coli o157:h7 isolates had at least two of the shiga-like toxin types i or ii or the attaching-effacing lesion g ...	1996	8789031
in vitro binding of type 1-fimbriated escherichia coli to uroplakins ia and ib: relation to urinary tract infections.	urinary tract infections, caused mainly by escherichia coli, are among the most common infectious diseases. most isolates of the uropathogenic e.coli can express type 1 and p fimbriae containing adhesins that recognize cell receptors. while p fimbriae recognize kidney glycolipid receptors and are involved in peyelonephritis, the urothelial for type 1 fimbriae were not identified. we show that type 1-fimbriated e. coli recognize uroplakins ia and ib, two major glycoproteins of urothelial apical p ...	1996	8790381
effect of a whey protein concentrate used as a colostrum substitute or supplement on calf immunity, weight gain, and health.	the efficacy of a whey protein concentrate was evaluated as a colostrum substitute or supplement in two experiments using four groups of 29 calves. in experiment 1, calves were fed either 2 l of pooled colostrum (group 1) or 500 g of whey protein concentrate (group 2). a mean total of 123.6 and 17.7 g of ig was fed to calves in groups 1 and 2, respectively. mean serum igg, total protein, and globulin concentrations and ig antibody activities to escherichia coli k99 and rotavirus were significant ...	1996	8792287
membrane sectors of f- and v-type h+-transporting atpases.	h+ transporting, reversible f-type atpases (atp synthases) and v-type atpases share major structural features and function by similar mechanisms. recent structural and genetic experiments provide new insights into the organization of the transmembrane and coupling sectors of the enzymes and the molecular mechanics of coupling h+ transport to atp.	1996	8794160
cloning and characterization of a xylanase gene from corn strains of erwinia chrysanthemi.	the gene encoding a 42-kda endoxylanase was cloned from erwinia chrysanthemi strain d1. sequencing of this gene, called xyna, showed that it encoded a primary protein product of 413 amino acids with an unusual and long (31 amino acid) leader peptide that was cleaved during secretion to the bacterial periplasm. this protein is distinct from xylanases in glycohydrolase families 10 and 11 and, instead, appears to be intermediate between families 5 and 30. the xyna gene is located downstream from a ...	1996	8810080
identification of histone h2a.x as a growth factor secreted by an androgen-independent subline of mouse mammary carcinoma cells.	shionogi carcinoma 115 (sc 115) cells and chiba subline 2 (cs 2) cells are clones of an androgen-responsive mouse tumor cell line and its autonomous subline, respectively. we have shown previously that cs 2 cells produce a heparin-binding growth factor that stimulates the growth of sc 115 cells as well as the growth of themselves. in this study, a growth factor was purified from serum-free conditioned media of cs 2 cells cultured without testosterone. a heparin-binding fraction showed growth- pr ...	1996	8810267
molecular epidemiology of escherichia coli o157:h7 by pulsed-field gel electrophoresis and comparison with that by bacteriophage typing.	one hundred twenty-four escherichia coli o157:h7 isolates were characterized by pulse-field gel electrophoresis, bacteriophage typing, and pcr of verotoxin genes. diversity indices obtained--0.786 for phage types and 0.987 for pulsed-field gel electrophoresis types--demonstrated that phage typing falls below the critical value (0.9) required for confident interpretation of results.	1996	8815116
naturally occurring acute coliform mastitis in holstein cattle.	physical examination and clinicopathologic findings from 44 adult holstein cows with naturally occurring coliform mastitis were studied. the cattle were grouped for comparison by stage of lactation and survival. cattle within the first 4 weeks of lactation maintained higher median mature neutrophil counts (1,200 versus 300/microl) in peripheral blood than cattle later in lactation. nonsurviving cows had higher median creatinine concentration (2.5 versus 1.6 mg/dl) and anion gap (25 versus 20 meq ...	1996	8819051
use of a modified bacteroides-prevotella shuttle vector to transfer a reconstructed beta-1,4-d-endoglucanase gene into bacteroides uniformis and prevotella ruminicola b(1)4.	a carboxymethyl cellulase (cmcase) gene from prevotella ruminicola b(1)4 was reconstructed by adding a cellulose binding domain from a thermomonospora fusca cellulase and was conjugally transferred from escherichia coli to bacteroides uniformis 0061 by using a chloramphenicol and tetracycline resistance shuttle vector (ptc-cow). ptc-cow was specifically constructed to facilitate conjugal transfer of vectors from b. uniformis donors to p. ruminicola recipients. b. uniformis transconjugants contai ...	1996	8572695
estimation of the h+/h- ratio of the reaction catalysed by the nicotinamide nucleotide transhydrogenase in chromatophores from over-expressing strains of rhodospirillum rubrum and in liposomes inlaid with the purified bovine enzyme.	two strains of rhodospirillum rubrum were constructed in which, by a gene dosage effect, the transhydrogenase activity of isolated chromatophores was increased 7-10-fold and 15-20-fold, respectively. the h+/h- ratio (the ratio of protons translocated per hydride ion equivalent transferred from nadph to an nad+ analogue, acetyl pyridine adenine dinucleotide), determined by a spectroscopic technique, was approximately 1.0 for chromatophores from the over-expressing strains, but was only approximat ...	1996	8573594
ribotyping to differentiate fusobacterium necrophorum subsp. necrophorum and f. necrophorum subsp. funduliforme isolated from bovine ruminal contents and liver abscesses.	differences in biological activities (hemagglutination, hemolytic, leukotoxic, and virulence) and ribotypes between the two subspecies of fusobacterium necrophorum of bovine ruminal and liver abscess origins were investigated. hemagglutination activity was present in all hepatic, but only some ruminal, strains of fusobacterium necrophorum subsp. necrophorum. ruminal f. necrophorum subsp. necrophorum had low leukotoxin titers yet was virulent in mice. fusobacterium necrophorum subsp. funduliforme ...	1996	8593050
unusual evolution of 11beta- and 17beta-hydroxysteroid and retinol dehydrogenases.	11beta-hydroxysteroid dehydrogenases regulate glucocorticoid concentrations and 17beta-hydroxysteroid dehydrogenases regulate estrogen and androgen concentrations in mammals. phylogenetic analysis of the sequences from two 11beta-hydroxysteroid dehydrogenases and four mammalian 17beta-hydroxysteroid dehydrogenases indicates unusual evolution in these enzymes. type 1 11beta- and 17beta-hydroxysteroid dehydrogenases are on the same branch; type 2 enzymes cluster on another branch with beta-hydroxy ...	1996	8593166
effect of vaccination with an escherichia coli bacterin-toxoid on milk production in dairy cattle.	to determine whether vaccination of lactating cattle with an escherichia coli j5 bacterin-toxoid would produce a significant short-term change in milk production.	1996	8837654
a cluster of escherichia coli o157:h7 infections with the hemolytic-uremic syndrome and death in california. a mandate for improved surveillance.	in mid-january 1993, an outbreak of escherichia coli o157:h7 infections associated with eating hamburger patties at a fast-food restaurant chain (chain a) was reported in washington state. from mid-december to mid-january, 9 cases of e coli o157:h7-associated bloody diarrhea and the hemolytic-uremic syndrome had been reported in san diego county, california. a total of 34 persons had bloody diarrhea, the hemolytic-uremic syndrome, or e coli o157:h7 organisms isolated from stool during the period ...	1996	8855679
molecular cloning of the cdna of mouse mitochondrial nadp-dependent isocitrate dehydrogenase and the expression of the gene during lymphocyte activation.	the current report documents the molecular cloning of the mouse mitochondrial nadp-dependent isocitrate dehydrogenase (mnadp-idh) cdna. the cdna was 1,863 bp in length and contained one open reading frame encoding a 523-residue polypeptide with a predicted molecular weight of 58 kda. the cdna and the deduced amino acid (aa) sequence of the mouse mnadp-idh had a high degree of homology with those of porcine, bovine, alfalfa, and yeast. the recombinant mnadp-idh expressed in escherichia coli had a ...	1996	8867815
genetic and immunological analyses of a 38 kda surface-exposed lipoprotein of pasteurella haemolytica a1.	pasteurella haemolytica serotype a1 is the bacterial pathogen most frequently isolated from the lungs of cattle with bovine respiratory disease. as part of a study to characterize p. haemolytica antigens which are important in eliciting resistance to pneumonic pasteurellosis, we have cloned and sequenced the gene encoding a 38 kda lipoprotein, lpp38. the deduced amino acid sequence of lpp38 is similar to those of the escherichia coli polyamine transport proteins potd (70%) and potf (33%). p. hae ...	1996	8870194
prevalence and characteristics of escherichia coli serotype o157:h7 and other verotoxin-producing e. coli in healthy cattle.	from february to july of 1994, 328 faecal samples from 32 herds were collected and verotoxin-producing escherichia coli (vtec) found on 84% of the farms. the proportion of animals infected varied from 0-63%. vtec were recovered from 52 (20%) of 257 cows and from 16 (23%) of 71 calves. although the vtec belonged to 25 different serogroups, 7 (o8, o20, o22, o77, o113, o126 and o162) accounted for 46% of strains. nearly 45% of the strains. nearly 45% of the 83 bovine vtec strains belonged to serogr ...	1996	8870622
structure and function in rhodopsin: expression of functional mammalian opsin in saccharomyces cerevisiae.	the yeast saccharomyces cerevisiae has been investigated for expression of mammalian opsin as an alternative to the currently used expression in cos-1 mammalian cells. the synthetic opsin gene was placed under the control of the inducible promoter gal1 in the multicopy yeast/ escherichia coli shuttle vector yeprf1. transformation of a gal+ s. cerevisiae strain with the vector and growth of galactose-induced cultures to saturation showed the production of 2.0 +/- 0.5 mg of opsin from about 10(10) ...	1996	8876161
emerging foodborne pathogens: escherichia coli o157:h7 as a model of entry of a new pathogen into the food supply of the developed world.	there would appear to be little argument that the large outbreaks of e. coli o157:h7 which have occurred since the early 1980s represent a distinct, new phenomenon. the number of reported cases have increased dramatically, starting from zero in 1981; however, it is also clear that this increase in reported cases is in part an artifact of improved surveillance and reporting. available data suggest that e. coli o157:h7 infections were present prior to 1982, although numbers appear to have been sma ...	1996	8877329
expression of carbohydrate-binding protein p33/41 in human tumor cell lines.	we previously reported a new type of lectin, p33/41 (annexin iv), which was isolated from a bovine tissue extract [kojima, k. et al. (1992) j. biol. chem. 267, 20536-20539]. when the expression of p33/41 (annexin iv) was surveyed in the lysates of 39 human tumor cell lines by sds-page, followed by western blot analysis with polyclonal anti-bovine p33/41 and monoclonal anti-annexin iv (z016, zymed) antibodies, 21 cell lines were found to be reactive with the polyclonal antibody, whereas all 39 ce ...	1996	8882729
incidence of clinical mastitis in a random sample of dairy herds in the southern netherlands.	the incidence of clinical mastitis and distribution of pathogens in dairy cows was estimated in 171 randomly selected dairy herds in the southern netherlands. a total of 1103 quarter cases were reported. the average annual incidence rate was 12.7 quarter cases per 100 cows per year. the most frequent isolates from clinical cases were escherichia coli (16.9 per cent), staphylococcus aureus (14.4 per cent), streptococcus uberis (11.9 per cent) and streptococcus dysgalactiae (8.9 per cent). most ca ...	1996	8883335
synthesis, processing, and oligomerization of bovine herpesvirus 1 ge and gi membrane proteins.	this study reports the identification and initial characterization of the precursors, modified forms, and oligomers of bovine herpesvirus 1 (bhv-1) gi and ge proteins with polyvalent rabbit serum specific for gi or ge. our experiments used the colorado strain of bhv-1 and mutant viruses with insertions of the escherichia coli lacz gene into the predicted ge and gi reading frames. we also translated the ge and gi open reading frames in vitro and expressed them in uninfected cells using eukaryotic ...	1996	8892910
detection of enterohaemorrhagic escherichia coli o157:h7 in minced beef using immunomagnetic separation.	enterohaemorrhagic escherichia coli (ehec) o157:h7 has been recently recognized as a human pathogen associated with haemorrhagic colitis and haemolytic uraemic syndrome. most outbreaks of haemorrhagic colitis resulted from the consumption of undercooked minced beef or raw milk. dairy cattle have been identified as a reservoir of ehec o157: h7. in this study e. coli o157 specific antibody, coated on magnetic beads, was used to concentrate and release ehec o157:h7 from meat samples. a survey of re ...	1996	8897418
in vitro renaturation of bovine beta-lactoglobulin a leads to a biologically active but incompletely refolded state.	when bovine beta-lactoglobulin (beta-lg) was refolded after extensive denaturation in 4.8 m guanidine hydrochloride (guhcl), the functional activity of the protein, retinol binding, as measured by the enhancement of this ligand's fluorescence, was completely recovered. in contrast, the room-temperature tryptophan phosphorescence lifetime of the refolded protein, a local measure of the residue environment, was approximately 10 ms, significantly shorter than the phosphorescence lifetime of the unt ...	1996	8897609
osteopontin n-terminal domain contains a cryptic adhesive sequence recognized by alpha9beta1 integrin.	osteopontin is an adhesive glycoprotein implicated in numerous diseases associated with inflammation and remodeling. there are several structural domains in osteopontin that are of particular interest. the rgd motif is a cell attachment sequence shown to be critical for cell adhesion through alphav-containing integrins. in close proximity to the rgd domain is the thrombin cleavage site. previous observations suggest that thrombin cleavage of osteopontin occurs in vivo and may be physiologically ...	1996	8910476
bacterial growth, inflammatory cytokine production, and neutrophil recruitment during coliform mastitis in cows within ten days after calving, compared with cows at midlactation.	to define causes of increased susceptibility to coliform mastitis after parturition.	1996	8915431
structure-biological activity relationships of 11-residue highly basic peptide segment of bovine lactoferrin.	the antimicrobial peptide, lactoferricin, is generated upon the gastric pepsin cleavage of lactoferrin and has many basic and hydrophobic amino acid residues essential for its biological activity. to investigate the structure-antimicrobial activity relationships, the basic amino acid-rich region of bovine lactoferricin (blfc), rrwqwrmkklg, was selected. using chemically synthesized blfc and its substituted peptides, the antimicrobial activities of the peptides were tested by determining the mini ...	1996	8919056
a pcr-based assay for the detection of escherichia coli shiga-like toxin genes in ground beef.	a detection system based on the pcr has been developed for escherichia coli strains which harbor the shiga-like toxin genes. this quantitative detection system involves the 5'-->3' nuclease activity of thermus aquaticus dna polymerase, which cleaves an internal oligonucleotide probe that has been labeled with both a fluorescent reporter dye (6-carboxy-fluorescein [fam]) and a quencher dye (6-carboxytetramethyl-rhodamine [tamra]). parameters which affected the performance of the assay included pr ...	1996	8919796
the restriction-modification system of pasteurella haemolytica is a member of a new family of type i enzymes.	genes encoding the type i restriction-modification (r-m) system of the bovine pathogen, pasteurella haemolytica, have been identified immediately downstream of a locus that encodes a transcriptional activator of p. haemolytica leukotoxin expression. type i enzymes are encoded by three genes called hsdm, hsds and hsdr, and have fallen into three groups, called ia, ib and ic. hsds provides a sequence recognition function which in concert with hsdm forms an active methyltransferase (mtase). inclusi ...	1996	8921897
equilibrium constants for nonspecific binding of proteins to dna may be obtained by quantitative zonal dna affinity chromatography.	quantitative zonal dna affinity chromatography may be used to determine accurate equilibrium constants for the binding of proteins nonspecifically to dna. zonal quantitative affinity chromatography has not previously been applied to the determination of binding constants of proteins to dna, although its use is quite commonplace for determination of affinity constants for protein-protein or protein-small ligand interactions. equilibrium constants were measured for the nonspecific binding of bovin ...	1996	8937566
cholesterol 7alpha-hydroxylase activities from human and rat liver are modulated in vitro posttranslationally by phosphorylation/dephosphorylation.	purified cholesterol 7alpha-hydroxylases (c7alphah) from human and rat liver microsomes, and from transformed escherichia coli expression systems, were incubated with 0.3 mmol/l [gamma-32p] adenosine triphosphate (atp) in the presence and absence of bacterial alkaline phosphatase (ap) or rabbit muscle adenosine 3',5'-cyclic monophosphate (camp)-dependent protein kinase. the amounts of 32p incorporation after separation of human and rat c7alphah proteins by sodium dodecyl sulfate-polyacrylamide g ...	1996	8938182
binding of an n-terminal rhodanese peptide to dnaj and to ribosomes.	a peptide corresponding to the n-terminal 17 amino acids of bovine rhodanese was fluorescently labeled with a coumarin derivative at its primary amino group(s) and then purified by high performance liquid chromatography. this peptide interacted with the molecular chaperone dnaj in the absence of other chaperones and atp. in the presence of atp, the molecular chaperone dnak bound to the dnaj-peptide complex, but not to the peptide alone. the chaperone grpe appeared to cause the release of the pep ...	1996	8940114
rapid and specific detection of f17-related pilin and adhesin genes in diarrheic and septicemic escherichia coli strains by multiplex pcr.	the f17-related adhesins are prevalent in escherichia coli strains isolated from calves with diarrhea or septicemia and from lambs with nephropathy. the f17 family includes the f17a, f17b, f17c, and f111 fimbriae produced by bovine e. coli strains and the g agglutinin produced by human uropathogenic e. coli strains. an easy and inexpensive multiplex pcr method was developed to detect all the f17-related fimbriae and to identify four subtypes of structural subunit genes and two distinct subfamili ...	1996	8940423
replication protein-a mediates the association of calf thymus dna polymerase alpha-dna primase complex with guanine-rich dna sequence.	we have shown that calf thymus dna polymerase alpha-dna primase complex (pol alpha-primase) preferentially binds to pyrimidine-rich sequences and initiates rna primer synthesis [suzuki, m. et al. (1993) biochemistry 32, 12782-12792]. here we tested the association of pol alpha-primase with a guanine-rich dna fragment (svg, 30-mer) containing in vivo initiation sites of simian virus 40 dna replication. while pyrimidine-rich fragment (ctpps 1, 30-mer), that is a preferred sequence for calf thymus ...	1996	8947839
human endothelial nitric oxide synthase: expression in escherichia coli, coexpression with calmodulin, and characterization.	human endothelial nitric oxide synthase (enos) has been cloned and expressed in escherichia coli. the spectroscopic properties and specific activity (100-130 nmol x min(-1) x mg(-1) at 37 degrees c) of the recombinant protein are similar to those of the bovine enzyme. fplc and low-temperature sds-page indicate that the protein is mostly dimeric in both the absence and presence of tetrahydrobiopterin. human enos thus has a higher tendency to dimerize than the bovine enzyme. a chloramphenicol-resi ...	1996	8951046
interpretive criteria for antimicrobial susceptibility testing of ceftiofur against bacteria associated with swine respiratory disease.	ceftiofur, an extended-spectrum cephalosporin, is active against a variety of animal pathogens, including organisms associated with swine respiratory disease. however, minimum inhibitory concentration (mic) breakpoint and disk diffusion interpretive criteria have not been established for swine pathogens. susceptibility tests were performed by broth microdilution mic and disk diffusion methods on 246 bacterial species that cause swine respiratory disease. ceftiofur was active against salmonella s ...	1996	8953533
rnas mediating cotranslational insertion of selenocysteine in eukaryotic selenoproteins.	selenocysteine, a selenium-containing analog of cysteine, is found in the prokaryotic and eukaryotic kingdoms in active sites of enzymes involved in oxidation-reduction reactions. its biosynthesis and cotranslational insertion into selenoproteins is performed by an outstanding mechanism, implying the participation of several gene products. the trna(sec) is one of these. in eukaryotes, its transcription mode by rna polymerase iii differs from that of classical trna genes, both at the level of the ...	1996	8955902
the atpase inhibitor protein from bovine heart mitochondria: the minimal inhibitory sequence.	the mitochondrial atpase inhibitor subunit is a basic protein of 84 amino acids that helps to regulate the activity of f1f0-atpase. in order to obtain structural information on the mechanism of inhibition, the bovine inhibitor subunit has been expressed in escherichia coli and purified in high yield. the recombinant protein has a similar inhibitory activity to the inhibitor subunit isolated from bovine mitochondria. progressive n-terminal and c-terminal deletion mutants of the inhibitor subunit ...	1996	8961923
[synthesis in escherichia coli and certain properties of hybrid protein consisting of a modified precursor of adrenodoxin reductase and shortened precursor of adrenodoxin].	some aspects of formation and functioning of the cholesterol hydroxylase system were studied. a hybrid protein was synthesized in e. coli composed of the modified form of the (nadph)adrenodoxin reductase precursor (n-terminal domain) and the shortened adrenodoxin precursor (c-terminal domain). the modified reductase precursor contained 12 extra amino acid residues at the n-terminus and the n-terminally shortened adrenodoxin precursor had 17 c-terminal amino acids of its targeting presequence. th ...	1996	8962919
the use of electrochemistry for the synthesis of 17 alpha-hydroxyprogesterone by a fusion protein containing p450c17.	a method has been developed for the commercial application of the unique oxygen chemistry catalyzed by various cytochrome p450s. this is illustrated here for the synthesis of hydroxylated steroids. this method requires the preparation of large amounts of enzymatically functional p450 proteins that can serve as catalysts and a technique for providing electrons at an economically acceptable cost. to generate large amounts of enzymatically active recombinant p450s we have engineered the cdnas for v ...	1996	8969926
the use of selective and differential agars in the isolation of escherichia coli o157 from dairy herds.	the use of selective and differential agars for the isolation of escherichia coli o157 from the faeces of dairy herds was investigated. out of the 614 isolates which were positive for one or more of the selective criteria employed in the media only four proved to be e. coli o157. ninety-nine per cent of the isolates were false positives. the procedure which resulted in the isolation of e. coli o157 from faecal samples was enrichment in modified tryptone soya broth supplemented with novobiocin an ...	1996	8972093
site-directed mutagenesis and spectroscopic characterization of human ferrochelatase: identification of residues coordinating the [2fe-2s] cluster.	the five cysteines closest to the carboxyl terminus of human ferrochelatase have been individually mutated to serine, histidine, or aspartate residues in an attempt to identify the protein ligands to the [2fe-2s] cluster. mutations of cysteines at positions 403, 406, and 411 (c403d, c403h, c406d, c406h, c406s, c411h, and c411s mutants) all resulted in inactive enzyme that failed to assemble the [2fe-2s] cluster as judged by whole-cell epr studies. in contrast, mutation of the cysteines at positi ...	1996	8973195
fimbriae extracts from enterotoxigenic escherichia coli strains of bovine and porcine origin with k99 and/or f41 antigens.	fimbriae extracts obtained using the thermal shock method, from bovine and porcine enterotoxigenic escherichia coli strains with k99 and/ or f41 antigens, were analyzed by sds-page, immunoblotting and haemagglutinating activity. three major protein bands with molecular weights 17 kda, 29.3 kda and 30.9 kda were detected depending on the strain assayed. a 17 kda band was identified as the fimbrial subunit for k99 fimbriae and was detected in strains of bovine and porcine origin. the 30.9 kda band ...	1996	9054120
[low molecular weight gtpase of the rac family from the bovine retina. structure of cdna and its expression in a prokaryotic system].	cdna with a high degree of homology to the rac1 gene of the ras-like protein from the hl-60 cell line was isolated from a bovine retina cdna library. the homologies of the nucleotide and deduced amino acid sequences were 90 and 98%, respectively. the product of cdna expression in e. coli cells purified by chromatography displayed gtp-binding and gtpase activities. the data suggest that the rac1 protein is one of the low-molecular gtp-binding proteins in photoreceptor membranes of vertebrates.	1996	9054337
influence of ozone on the susceptibility of escherichia coli k1 to the bactericidal action of serum.	the susceptibility to the bactericidal action of normal bovine serum of twenty two escherichia coli k1 strains, isolated from the urine of patients with urinary tract infections, was determined. only four strains were resistant. ozonization of bacterial suspensions enhanced the sensitivity of the strains to the action of both normal serum and a serum in which the alternative pathway of complement activation was thermally blocked.	1996	9131796
thermal activation of the bovine hsc70 molecular chaperone at physiological temperatures: physical evidence of a molecular thermometer.	differential scanning calorimetry was used to monitor the thermal transitions of the 70 kda heat shock cognate protein (hsc70). hsc70 had endothermic transitions with midpoints (tm) at 59 degrees c and 63 degrees c in the absence and presence of atp, respectively, and a similar increase in tm was observed using intrinsic fluorescence of tryptophan. combined with increased exposure at 60 degrees c of non-polar residues of hsc70 to which the hydrophobic, fluorescent probe ans bound, these data ind ...	1996	9222592
immunochemical detection of adp-ribosylating enzymes in the archaeon sulfolobus solfataricus.	polyclonal antibodies raised against eukaryotic mono-(adpribose)transferase and poly(adpribose)polymerase were used to test the presence of antigenic determinants in a crude extract of sulfolobus solfataricus, a thermophilic archaeon. samples from eukaryotic (bull testis) and bacterial (e. coli) sources were analysed for comparison. all tested antibodies reacted with the sulfolobal sample with a specificity comparable to that of the eukaryotic preparation, as revealed by elisa test, activity ass ...	1996	8549833
isoleucine 368 is involved in low-affinity binding of n6-modified camp analogues to site b of the regulatory subunit of camp-dependent protein kinase i.	the regulatory (r) subunit of camp-dependent protein kinase has a well-defined domain structure including the two in-tandem camp-binding sites that constitute the c-terminus of the protein. the n-terminal binding site (a) has a considerably higher affinity for analogues of camp that are substituted with bulky and hydrophobic substituents at the 6-amino group of the adenine ring compared to the affinity observed at the second site (b). on the basis of the crystal structure of the catabolite gene ...	1996	8645227
cloning, sequencing and expression of the bovine cd3 epsilon and tcr-zeta chains, two invariant components of the t-cell receptor complex.	cd3 epsilon and the zeta-chain of the bovine t-cell receptor (tcr) are two invariant molecules with an important role in signal transduction via the tcr/cd3 complex. the nucleotide sequence of a bovine cd3 epsilon cdna clone containing the complete coding sequence was determined and the deduced amino acid (aa) sequence compared to that of other species. the cytoplasmic domains of the different cd3 epsilon clearly show a higher degree of conservation than the extracellular domains. bovine cd3 eps ...	1996	8647441
complementary dna cloning of the predominant allergen of bovine dander: a new member in the lipocalin family.	a number of allergenic proteins in animal danders have been characterized at the molecular level, but little is known of their biologic functions. we have found that the prevalence of ige antibodies among patients with cattle-associated asthma is highest against a dander protein referred to as bda20.	1996	8648026
enterohaemorrhagic escherichia coli: a new problem, an old group of organisms.	all mammals are colonised by escherichia coli generally at birth and these organisms become part of their intestinal flora for the rest of their lives. new types are acquired generally by an oral route. some e coli are pathogenic and some may have a far more enhanced ability to colonise the human intestine than most others. recently enterohaemorrhagic e coli have emerged. they can cause a number of intestinal illnesses in humans including bloody diarrhoea and haemolytic uraemic syndrome. these o ...	1996	8660186
inhibition of nad+ glycohydrolase and adp-ribosyl cyclase activities of leukocyte cell surface antigen cd38 by gangliosides.	we have recently reported that gangliosides act as inhibitors of adp-ribosyltransferases and nad+ glycohydrolases (nadase) of pertussis toxin and the c3 exoenzyme from clostridium botulinum (hara-yokoyama, m., hirabayashi, y., irie, f., syuto, b., moriishi, k., sugiya, h., and furuyama, s. (1995) j. biol. chem. 270, 8115-8121). here, we investigated the effect of gangliosides on the enzymatic activity of leukocyte cell surface antigen cd38, which is identified as an ecto-nadase (kontani, k., nis ...	1996	8662799
identification of topoisomerase i as the cytotoxic target of the protoberberine alkaloid coralyne.	protoberberine alkaloids (coralyne and its derivatives), which exhibit antileukemic activity in animal models, have been shown to be potent inducers of topoisomerase (topo) i-dna cleavable complexes using purified recombinant human dna topo i. different from the structurally similar benzophenanthridine alkaloid nitidine (a dual poison of both topos i and ii), coralyne and its derivatives have marginal poisoning activity against dna topo ii. yeast cells expressing human dna topo i are shown to be ...	1996	8665516
the delta- and epsilon-subunits of bovine f1-atpase interact to form a heterodimeric subcomplex.	the delta-subunit of bovine f1-atpase was expressed from a bacterial vector at fairly high level in escherichia coli, but the yield of bovine epsilon-subunit was rather low under similar conditions. however, co-expression of the proteins from a dicistronic operon delta-epsilon in the same expression vector, produced both of them in good yield in a soluble form in the bacterial cytoplasm, and by chromatography it was found that the delta- and epsilon-subunits were associated in a stable complex. ...	1996	8670087
alpha-tocopherol concentrations in milk and plasma during clinical escherichia coli mastitis.	eighteen cows were challenged by intramammary infusion with escherichia coli 727 to determine the effects of acute clinical mastitis on alpha-tocopherol concentrations in plasma and milk. cows were fed diets supplemented with 1000 iu of vitamin e/d from calving through the experimental period. at challenge, geometric mean dim was 33 d. each mammary quarter was diagnosed with an imi and clinical mastitis at 24 and 48 h after challenge. the alpha-tocopherol concentrations in milk from challenged q ...	1996	8675784
native complex formation between apolipoprotein e isoforms and the alzheimer's disease peptide a beta.	to explore whether the genetic linkage between apolipoprotein e (apoe) alleles and susceptibility to alzheimer's disease might be attributable to a direct molecular interaction between apoe and the amyloid peptide a beta, we have produced apoe variants in escherichia coli and studied their interactions with a beta under native conditions. when incubated with a beta at 20-40 microm concentrations, all three isoforms of apoe (2, 3, and 4) readily form complexes with a beta which can be isolated by ...	1996	8679539
gabexate mesilate, a synthetic protease inhibitor, attenuates endotoxin-induced pulmonary vascular injury by inhibiting tumor necrosis factor production by monocytes.	in order to determine whether gabexate mesilate, a synthetic protease inhibitor with anticoagulant properties, is useful for the treatment of adult respiratory distress syndrome, we examined its effect on endotoxin-induced pulmonary vascular injury in rats.	1996	8681572
quantification of staphylococcus aureus and escherichia coli in the liquid medium by fluorimetry and its use in phagocytosis assay.	a fluorimetric technique was compared with the plate counting method for quantification of viable cells of staphylococcus aureus and escherichia coli in the liquid medium. the fluorimetric assay measures the release of fluorogenic 4-methylumbelliferone (4-mu) from 4-methylumbelliferyl phosphate by the bacterial phosphatases. the increase in fluorescence was dependent on the size of bacterial inocula. setting the fluorescence threshold at the middle of the logarithmic growth phase resulted in goo ...	1996	8698658
rapid physicochemical detachment, separation and concentration of bacteria from beef surfaces.	a simple, rapid, physicochemical treatment for the removal of viable bacteria from the surface of raw and cooked beef is described. the detachment method was linked to a differential centrifugation step which removed large amounts of particulate food matter and concentrated the detached bacteria. the method increased the numbers of bacteria released from beef surfaces and increased the numbers detected by at least one and a half orders of magnitude, when compared to the traditional 'stomaching' ...	1996	8698669
s-nitrosoglutathione is cleaved by the thioredoxin system with liberation of glutathione and redox regulating nitric oxide.	in activated human neutrophils a burst of nitric oxide (no) converts intracellular gsh to s-nitrosoglutathione (gsno) which is subsequently cleaved to restore gsh by an unknown mechanism. we discovered that gsno is an nadph oxidizing substrate for human or calf thymus thioredoxin reductase (tr) with an apparent km value of 60 microm and a kcat of 0.6 x s-1. addition of human thioredoxin (trx) stimulated the initial nadph oxidation rate severalfold but was accompanied by progressive inactivation ...	1996	8702596
membrane topography and near-neighbor relationships of the mitochondrial atp synthase subunits e, f, and g.	the well characterized subunits of the bovine atp synthase complex are the alpha, beta, gamma, delta, and epsilon subunits of the catalytic sector, f1; the atpase inhibitor protein; and subunits a, b, c, and d, oscp (oligomycin sensitivity-conferring protein), f6, and a6l, which are present in the membrane sector, f0, and the 45-a-long stalk that connects f1 to f0. it has been shown recently that bovine atp synthase preparations also contain three small polypeptides, designated e, f, and g, with ...	1996	8702768
inhibitor probes of the quinone binding sites of mammalian complex ii and escherichia coli fumarate reductase.	the structural and catalytic properties of beef heart succinate dehydrogenase (succinate-ubiquinone oxidoreductase, complex ii) and escherichia coli fumarate reductase are remarkably similar. one exception is that whereas electron exchange between the mammalian enzyme and its quinone pool is inhibited by thenoyltrifluoroacetone and carboxanilides, the enzyme from e. coli is not sensitive to these inhibitors. the lack of good inhibitors has seriously hampered the elucidation of the mechanism of q ...	1996	8702865
cloning and expression of human mitochondrial deoxyguanosine kinase cdna.	mammalian mitochondrial deoxyguanosine kinase (dgk) is responsible for phosphorylation of purine deoxyribonucleosides in the mitochondrial matrix. using a rt-pcr-generated probe, based on amino acid sequence information from proteolytic fragments of purified bovine dgk, we have cloned a cdna from a human brain cdna library that encodes a 30 kda protein. the deduced amino acid sequence of this protein included the sequence of all six peptides isolated and sequenced from purified dgk. expression a ...	1996	8706825
effects of freezing on the viability of nine pathogens from quarters with subclinical mastitis.	milk samples from 45 quarters containing mastitis pathogens were collected from lactating cows to determine the viability of those pathogens after freezing. an initial bacteria count was conducted, and samples were divided into 2-ml portions and frozen. weekly bacteria counts were conducted for 6 wk. viability after freezing was determined on five isolates of nine bacterial species: staphylococcus aureus, staphylococcus hyicus, staphylococcus chromogenes, staphylococcus xylosus, streptococcus ag ...	1996	8708092
[enterohaemorrhagic escherichia coli o157:h7 infection].	escherichia coli o157:h7 differs from previously described diarrheagenic e. coli classes (enteropathogenic, enteroinvasive, enterotoxigenic) by distinct clinical symptoms, production of verotoxin (vt) and a specific plasmid. cattle are the primary reservoirs of e. coli o157:h7. the organism may be transmitted through the consumption of contaminated foods (mainly of bovine origin) and by person-to-person contact. the most typical clinical manifestations of e. coli o157:h7 infection are hemorrhagi ...	1996	8710429
intramammary defense against infections induced by escherichia coli in cows.	to examine escherichia coli lipopolysaccharide (lps) effects on expression of cd14 and cd18 cell surface receptors and lectin/carbohydrate-mediated nonopsonic phagocytosis of e coli.	1996	8712510
efficacy of florfenicol for treatment of clinical and subclinical bovine mastitis.	to evaluate efficacy of florfenicol treatment for bovine mastitis caused by streptococcus agalactiae, staphylococcus aureus nonagalactiae streptococci, coagulase-negative staphylococci, escherichia coli, klebsiella sp, and others.	1996	8712519
biochemical and immunological identification and enrichment of poly(a) polymerase from human thymus.	human thymus poly(a) polymerase (ec 2.7.7.19) activity has been investigated using poly(a) and oligo(a) as initiators. all obtained fractions reveal more than one polypeptide as detected by immunoblotting after sds-page. in addition to the homogeneously purified (tsiapalis et al., j biol chem 250: 4486-4496, 1975 and wahle, j biol chem 266: 3131-3139, 1991), about 60 kda polypeptide, a larger polypeptide, about 80 kda, that comigrates in the region of poly(a) polymerase activity was detected, en ...	1996	8717411
epidemiological characteristics of bovine clinical mastitis caused by staphylococcus aureus and escherichia coli studied by dna fingerprinting.	to study the epidemiology of clinical mastitis caused by escherichia coli and staphylococcus aureus by differentiating isolates with dna fingerprinting techniques, using polymerase chain reaction.	1996	8720235
chemosensory responses of acanthamoeba castellanii: visual analysis of random movement and responses to chemical signals.	a visual assay slide chamber was used in conjunction with time-lapse videomicroscopy to analyze chemotactic behavior of axenically grown acanthamoeba castellanii. data were collected and analyzed as vector scatter diagrams and cell tracks. amebas responded to a variety of bacterial products or potential bacterial products by moving actively toward the attractant. responses to the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (fmlp), lipopolysaccharide, and lipid a were statistically ...	1996	8720945
sensitization of bacteria to danofloxacin by temperate prophages.	danofloxacin (cp-76,136) is in a class of agents that inhibit dna gyrase and trigger induction of the sos response and temperate bacteriophages. killing studies against the bovine pathogen pasteurella haemolytica demonstrated that danofloxacin exhibits particularly rapid killing kinetics. here, lysogenic escherichia coli bearing lambda is found to be more sensitive to danofloxacin than nonlysogenic e. coli. danofloxacin exposure also induced a prophage in p. haemolytica. the potency of danofloxa ...	1996	8726041
bacterial lipopolysaccharide-stimulated nitric oxide generation is unrelated to concurrent cytotoxicity of bovine alveolar macrophages.	nitric oxide (no) is a reactive metabolite produced by stimulated macrophages, and it has been demonstrated to exert cytotoxic actions on a number of microbes, parasites, and tumor cells. in addition, no has been reported to have an autotoxic effect on murine macrophages, its site of synthesis. we have investigated the relationship of no generation to cytotoxicity of bovine alveolar macrophages (am) in vitro, and have also assessed the effects of several modulators of cellular function on this r ...	1996	8728020
the infectious bronchitis virus nucleocapsid protein binds rna sequences in the 3' terminus of the genome.	the infectious bronchitis virus (ibv) nucleocapsid protein was expressed as a bacterial fusion protein which differed from the native protein only in the addition of six amino terminus histidine residues. using rna overlay protein blot assays, the recombinant protein was shown to bind to rna fragments specific for the positive sense 3' noncoding end of the ibv genome. at greater concentrations of sodium chloride, the native and fusion nucleocapsid proteins similarly bound to g rna, representing ...	1996	8599203
characterization of bovine endothelial nitric oxide synthase expressed in e. coli.	bovine endothelial constitutive nitric oxide synthase (enos) was expressed in e. coli as a soluble, catalytically active enzyme using the pcw expression vector coexpressed with a plasmid, pgroels, encoding the chaperonins groel and groes. the e. coli bl21 cultures reproducibly synthesized 6-10 mg of recombinant enzyme per liter of culture. the enos protein was purified using 2'5'-adp sepharose 4b and appeared as a single band of apparent molecular mass 135 kda on sds/page. the recombinant restin ...	1996	8604992
the role of the insertion loop around tryptophan 148 in tthe activity of thrombin.	thrombin has trypsin-like specificity for arg-xaa and lys-xaa peptide bonds; however, it is much more specific than trypsin, cleaving far fewer peptide bonds in macromolecular substrates. to probe the nature of the specificity of thrombin, a mutant has been constructed in which the trp148 loop of thrombin has been replaced with the same loop of bovine trypsin. this mutant was expressed in escherichia coli as prethrombin-2(148) using a t7 expression system previously described for wild-type preth ...	1996	8605192
characterization of the active intermediate of a groel-groes-mediated protein folding reaction.	recent studies of groe-mediated protein folding indicate that substrate proteins are productively released from a cis ternary complex in which the nonnative substrate is sequestered within the groel channel underneath groes. here, we examine whether protein folding can occur in this space. stopped-flow fluorescence anisotropy of a pyrene-rhodanese-groel complex indicates that addition of groes and atp (but not adp) leads to a rapid change in substrate flexibility at groel. strikingly, when groes ...	1996	8608602
purification of a mammalian homologue of escherichia coli endonuclease iii: identification of a bovine pyrimidine hydrate-thymine glycol dnase/ap lyase by irreversible cross linking to a thymine glycol-containing oligoxynucleotide.	we purified a homologue of the escherichia coli dna repair enzyme endo nuclease iii 5000-fold from calf thymus which, like endonuclease iii, demonstrates dna-glycosylase activity against pyrimidine hydrates and thymine glycol and ap lyase activity (dna strand cleavage at ap sites via beta-elimination). the functional similarity between the enzymes suggested a strategy for definitive identification of the bovine protein based on the nature of its enzyme-substrate (es) intermediate. prokaryotic dn ...	1996	8611553
pathogenesis of corneal infection: binding of pseudomonas aeruginosa to specific phospholipids.	clinical isolates of pseudomonas aeruginosa were examined for binding interactions with phospholipids of corneal epithelium. thin-layer chromatography (tlc) of lipids extracted from corneal epithelia followed by staining with an ammonium molybdate spray reagent revealed three phospholipid components, pl1, pl2, and pl3. the chromatographic mobility of pl1 was similar to that of the phospholipid standards phosphatidylinositol (pi) and phosphatidylserine (ps), which were not well resolved from one ...	1996	8613396
antagonism of lactic acid bacteria towards staphylococcus aureus and escherichia coli on agar plates and in milk.	the antagonistic effect of lactic acid bacteria (lab, including lactobacillus acidophilus, l. bulgaricus, l. casei and streptococcus thermophilus) on staphylococcus aureus and escherichia coli was evaluated on mrs agar with the deferred and cross-streaking techniques, and in milk with the plate counting method. all lab were repressive to s aureus and e coli on the agar medium. however, their suppressive activity was significantly reduced when the agar medium was buffered to ph 7.2. in normal mil ...	1996	8620186
physiological responses to repeated endotoxin challenge are selectively affected by recombinant bovine somatotropin administration to calves.	the study determined 1) whether the pretreatment of calves with recombinant bovine somatotropin (bst, sometribove) would alter the change in packed cell volume (pcv), rectal temperature (rt), and the plasma concentrations of ca2+, fe2+, glucose (g), urea n (pun), nonesterified free fatty acids (nefa), albumin (alb), and blood cell populations after endotoxin challenge (ec) and 2) whether the natural development of physiologic tolerance to repeated ec was affected by bst. twelve steer beef calves ...	1996	8625619
prevalence of the eaea gene in verotoxigenic escherichia coli strains from dairy cattle in southwest ontario.	this study determined the prevalence of the eaea gene and its relationship to serotype and type of verotoxin produced in a collection of 432 verotoxigenic escherichia coli (vtec) obtained from the faeces of healthy cows and calves in a systematic random survey involving 80 dairy farms in southwest ontario. a pcr amplification procedure involving primer pairs which target the conserved central region of the o157:h7 eaea gene showed that 151 (35.2%) strains were positive for the eaea gene. all iso ...	1996	8625998
assessment of the atp binding properties of hsp90.	hsp90, one of the most prominent proteins in eucaryotic cells under physiological and stress conditions, chaperones protein folding reactions in an atp-independent way. surprisingly, atp binding and atpase activity of hsp90 has been reported by several groups. to clarify this important issue, we have reinvestigated the potential atp binding properties and atpase activity of highly purified hsp90 using a number of different techniques. hsp90 was compared to the well characterized atp-binding chap ...	1996	8626558
endothelial nitric-oxide synthase. expression in escherichia coli, spectroscopic characterization, and role of tetrahydrobiopterin in dimer formation.	bovine endothelial nitric-oxide synthase (enos) expressed in escherichia coli does not have the post-translational modifications found in the native enzyme and is free of tetrahydrobiopterin (bh4). in the presence of bh4, enos has an absorption maximum at 400 nm that shifts to 395 nm when the substrate l-arginine is added. the low-spin component of the spectrum of the bh4-free protein is decreased by the addition of bh4 without a corresponding increase in the high-spin component. addition of bh4 ...	1996	8626704
the carboxyl-terminal domain (111-165) of vascular endothelial growth factor is critical for its mitogenic potency.	vascular endothelial growth factor (vegf) is a potent and specific mitogen for endothelial cells. vegf is synthesized and secreted by many differentiated cells in response to a variety of stimuli including hypoxia. vegf is expressed in a variety of tissues as multiple homodimeric forms (121, 165, 189, and 206 amino acids/monomer) resulting from alternative rna splicing. vegf121 is a soluble mitogen that does not bind heparin; the longer forms of vegf bind heparin with progressively higher affini ...	1996	8631822
eukaryotic protein disulfide isomerase complements escherichia coli dsba mutants and increases the yield of a heterologous secreted protein with disulfide bonds.	eukaryotic protein disulfide isomerase (pdi) is a 55-kda enzyme with cysteine oxidoreductase, chaperone, and antichaperone activities that catalyzes disulfide formation and rearrangement in the eukaryotic endoplasmic reticulum. in gram-negative bacteria, the formation of disulfide bonds in the periplasm is mediated by dsba, a strong cysteine oxidase but an inefficient catalyst of disulfide bond isomerization with no known chaperone activity. we show that rat pdi (rpdi) secreted in the periplasmi ...	1996	8631865
prevalence and clonal nature of escherichia coli o157:h7 on dairy farms in wisconsin.	a survey was conducted between march and october of 1994 to determine the prevalence and identify the sources of serotype o157:h7 isolates of escherichia coli in wisconsin dairy herds. a stratified sample of 400 farms was identified, and 70 farms with weaned calves less than 4 months old were included in the study. during the prevalence study, 5 of the 70 farms (herd prevalence, 7.1 +/- 4.5%) and fecal samples from 10 of 560 calves (animal prevalence, 1.8%) tested positive for serotype o157:h7. ...	1996	8633851
nucleotide sequence and expression in e. coli of the complete p4 type vp4 from a g2 serotype human rotavirus.	the complete sequence of a p4 type vp4 gene from a g2 serotype human rotavirus, is2, isolated in india has been determined. although the is2 vp4 is highly homologous to the other p4 type alleles, it contained acidic amino acid substitutions at several positions that make it acidic among the p4 type alleles that are basic. moreover, comparative sequence analysis revealed unusual polymorphism in members of the p4 type at amino acid position 393 which is highly conserved in members of other vp4 typ ...	1996	8634023
the processing proteases prohormone thiol protease, pc1/3 and pc2, and 70-kda aspartic proteinase show preferences among proenkephalin, proneuropeptide y, and proopiomelanocortin substrates.	proteases of cysteine, aspartic, and subtilisin classes have been indicated as candidate prohormone processing enzymes. the chromaffin granule proenkephalin processing proteases have been characterized as the novel cysteine protease prohormone thiol protease (ptp), a 70-kda aspartic proteinase, and the subtilisin-like pc1/3 and pc2 enzymes. the goal of this study was to assess whether these processing proteases possess preference(s) for prohormone substrates. the recombinant prohormones proenkep ...	1996	8638918
carboranyl oligonucleotides. 3. biochemical properties of oligonucleotides containing 5-(o-carboranyl-1-yl)-2'-deoxyuridine.	boronated oligonucleotides are potential candidates for boron neutron capture therapy, antisense technology, and as tools in molecular biology. the biological properties of dodecathymidylic acids containing one or more 5-(o-carboran-1-yl)-2'-deoxyuridine residues at different locations within the oligonucleotide chain were studied. 5-(o-carboran-1-yl)-2'-deoxyuridine containing oligonucleotides manifested marked increased lipophilicity and resistance to 3'- or 5'-phosphodiesterases compared to t ...	1996	8639534
glycans of bovine lactoferrin function as receptors for the type 1 fimbrial lectin of escherichia coli.	bovine lactoferrin strongly inhibited the hemagglutination activity of type 1 fimbriated escherichia coli. in addition, it agglutinated these bacteria. the agglutination reaction was specifically inhibited by glycopeptides derived from bovine lactoferrin or alpha-methyl-d-mannoside. these observations indicate that the glycans of bovine lactoferrin can serve as receptors for type 1 fimbrial lectin.	1996	8641764