Publications

phospholipase a2 engineering. x-ray structural and functional evidence for the interaction of lysine-56 with substrates.	site-directed mutagenesis studies of bovine pancreatic phospholipase a2 (pla2, overproduced in escherichia coli) showed that replacement of surface residue lys-56 by a neutral or hydrophobic amino acid residue resulted in an unexpected and significant change in the function of the enzyme. the kcat for phosphatidylcholine micelles increases 3-4-fold for k56m, k56i, and k56f and ca. 2-fold for k56n and k56t but does not change for k56r. these results suggest that the side chain of residue 56 has s ...	1991	1751497
[synthesis of the e-antigen of the hepatitis b virus (hbeag) in eukaryotic cells by a recombinant strain of the vaccinia virus].	the recombinant plasmids containing the gene for hepatitis b viral core-antigen with the pre-core-sequence controlled by the early-late promoter of the 7.5' k protein gene were constructed. the recombinant strains of vaccinia virus were obtained on their basis (vhbe42-1 and vhbe42-3) selectively expressing hbeag of hepatitis b virus. the kinetics of hbeag synthesis was studied in infected cells as well as secretion of the protein into culturing medium. three proteins were found by blotting techn ...	1991	1745274
stimulation of dna polymerase alpha activity by microtubule-associated proteins.	microtubule-associated protein 2 (map2) isolated from porcine brains stimulated the activity of dna polymerase alpha immunopurified from calf thymus or human lymphoma cells, in a dose-dependent manner. this stimulation was pronounced when activated dna or poly(da).(dt)10 was used as the template-primer. dna polymerase alpha bound to a map2-immobilized column, whereas preincubation of the enzyme with unbound map2 prevented binding to the column. these events suggested that a physical binding occu ...	1991	1742279
cloning and expression in escherichia coli of mature e1 beta subunit of bovine mitochondrial branched-chain alpha-keto acid dehydrogenase complex. mapping of the e1 beta-binding region on e2.	a cdna encoding the mature e1 beta subunit of the bovine branched-chain alpha-keto acid dehydrogenase complex was isolated from a lambda zap expression library. the bovine e1 beta cdna is 1,393 base pairs in length. it encodes the entire mature e1 beta subunit consisting of 342 amino acid residues and a partial mitochondrial targeting presequence of 26 residues. the calculated molecular mass of the mature bovine e1 beta subunit is 37,776 daltons, and the calculated isoelectric point is pi 5.04. ...	1992	1730724
the g226a mutant of gs alpha highlights the requirement for dissociation of g protein subunits.	adenylylcyclase cannot be activated by hormones or guanine nucleotide analogs in membranes from cells that express the g226a mutant form gs alpha instead of the wild-type protein. the mutant gs alpha protein appears incapable of undergoing the conformational change necessary for guanine nucleotide-induced dissociation of the g protein alpha subunit from the beta gamma subunit complex (miller, r.t., masters, s.b., sullivan, k.a., beiderman, b., and bourne, h.r. (1988) nature 334, 712-715). g226a ...	1992	1730644
cloning, sequencing, expression, and functional studies of a 15,000-molecular-weight haemophilus somnus antigen similar to escherichia coli ribosomal protein s9.	haemophilus somnus is a gram-negative bacterium capable of causing a number of disease syndromes in cattle. this article describes the cloning and characterization of a gene coding for a 15,000-molecular-weight (15k) polypeptide which reacts strongly with antiserum against h. somnus. analysis of plasmid-encoded polypeptides by polyacrylamide gel electrophoresis showed that the corresponding gene is the second in a transcriptional unit. the first gene codes for a protein with a molecular weight o ...	1992	1729207
nitric oxide from vascular smooth muscle cells: regulation of platelet reactivity and smooth muscle cell guanylate cyclase.	1. incubation of smooth muscle cells (smc) from bovine aorta for 3 min with human washed platelets treated with indomethacin (10 microm) promoted a cell number-related inhibition of platelet aggregation induced by thrombin (40 mu ml-1). this inhibition was not attributable to products of the cyclo-oxygenase pathway for the smc were also treated with indomethacin (10 microm). 2. the inhibitory activity of the smc on platelet aggregation was enhanced by incubating the smc with e. coli lipopolysacc ...	1991	1724627
swine and cattle enterotoxigenic escherichia coli-mediated diarrhea. development of therapies based on inhibition of bacteria-host interactions.	enterotoxigenic escherichia coli (etec) frequently occurs in diarrheal disease afflicting domestic animals. in this paper is summarized the research carried out over the last decade on the two important determinants of virulence that plays a role in the development of the infection, namely the colonizing ability of the small intestine mediated by specific fimbrial adhesins acting as lectins and the production of enterotoxins. recent progress in knowledge of the phenomenon led to alternative stra ...	1991	1723693
glycosylation of insulin-like growth factor binding protein-3 (igfbp-3) is not required for potentiation of igf-i action: evidence for processing of cell-bound igfbp-3.	insulin-like growth factor binding protein-3 (igfbp-3) is unique among the igf binding proteins in its extensive glycosylation in the native state. to determine the functional significance of carbohydrate moieties on igfbp-3, we examined the effects of nonglycosylated escherichia coli-derived recombinant human igfbp-3 (higfbp-3e. coli) and glycosylated chinese hamster ovary cell-derived higfbp-3 (higfbp-3cho) on igf-i action in cultured bovine fibroblasts. both higfbp-3 preparations bound igf-i ...	1991	1720092
isolation of a cdna encoding a mammalian multiubiquitinating enzyme (e225k) and overexpression of the functional enzyme in escherichia coli.	the ubiquitin (ub)-conjugating enzyme e2(25k) catalyzes the synthesis of multi-ub chains in which successive ub units are linked by an isopeptide bond involving the epsilon-amino group of lys-48 of ubn, and the cooh-terminal gly residue of ubn+1 (chen, z., and pickart, c. m. (1990) j. biol. chem., 265, 21835-21842). we now describe the polymerase chain reaction (pcr)-based cloning of an e2(25k)-encoding cdna from a bovine thymus library, using degenerate oligonucleotide primers based on the sequ ...	1991	1714895
hp 0.35, a cephalosporin degradation product is a specific inhibitor of lentiviral rnases h.	penicillins, cephalosporins and other betalactam antibiotics are widely used antibacterial drugs. recently it was found that some of them also have effects on proliferating eukaryotic cells (neftel, k.a. and hübscher, u. (1987) antimicrob. agents chemother. 31, 1657-1661), and one such effect was shown to be the inhibition of dna polymerase alpha (huynh do,u., neftel, k.a., spadari, s. and hübscher, u. (1987) nucl. acids res. 15, 10495-10506). the data suggested that degradation products of beta ...	1991	1714562
molecular cloning and expression of biologically active human glia maturation factor-beta.	glia maturation factor-beta, a protein found in the brains of all vertebrates thus far examined, appears to play a role in the differentiation, maintenance, and regeneration of the nervous system. using oligonucleotide probes based on the sequences of three tryptic peptides derived from bovine glia maturation factor-beta, we screened a human brainstem cdna library in lambda gt11. a 0.7-kb clone was isolated, sequenced in its entirety, and found to encode a polypeptide of 142 amino acids which co ...	1991	1712830
cloning, expression and characterization of the human transcription elongation factor, tfiis.	the cdna for the human elongation factor, tfiis, has been cloned and expressed in e. coli with the t7 expression system. this 280-amino acid tfiis protein is shorter by 21 residues than that of the mouse. the missing 21 residues are located in the amino-terminal region, which is not thought to be required for transcriptional stimulation. apart from this gap, human and mouse proteins reveal 96% overall identity and 98.5% sequence similarity if conservative substitutions are taken into account. th ...	1991	1708494
covalent binding studies on the 14c-labeled antitumor compound 2,5-bis(1-aziridinyl)-1,4-benzoquinone. involvement of semiquinone radical in binding to dna, and binding to proteins and bacterial macromolecules in situ.	2,5-bis(1-aziridinyl)-1,4-benzoquinone (babq) is a compound from which several antitumour drugs are derived, such as trenimone, carboquone and diaziquone (azq). the mechanism of dna binding of babq was studied using 14c-labeled babq and is in agreement with reduction of the quinone moiety and protonation of the aziridine ring, followed by ring opening and alkylation. the one-electron reduced (semiquinone) form of babq alkylates dna more efficiently than two-electron reduced or non reduced babq. ...	1990	1699678
denaturant-dependent folding of bovine pancreatic trypsin inhibitor mutants with two intact disulfide bonds.	the equilibrium and kinetic behavior of the guanidine hydrochloride (gdn-hcl) induced unfolding/refolding of four bovine pancreatic trypsin inhibitor (bpti) mutants was examined by using ultraviolet difference spectroscopy. in three of the mutants, we replaced the buried 30-51 disulfide bond with alanine at position 51 and valine (val30/ala51), alanine (ala30/ala51), or threonine (thr30/ala51) at position 30. for the fourth mutant, the solvent-exposed 14-38 disulfide was substituted by a pair of ...	1990	1693524
a genetic screen to identify variants of bovine pancreatic trypsin inhibitor with altered folding energetics.	a genetic screening procedure has been developed to identify mutant forms of bovine pancreatic trypsin inhibitor (bpti) that can fold to an active conformation but are inactivated more rapidly than the wild-type protein. small cultures of escherichia coli containing plasmids with mutagenized bpti genes were grown in microtiter plates, lysed, and treated with dithiothreitol (dtt). under these conditions, unfolding and inactivation of the wild-type protein has a half-time of about 10 hours. varian ...	1990	1691852
expression of bovine leukaemia virus antigens fused to ms2 polymerase in e. coli.	the main core protein and segments of envelope proteins of bovine leukaemia virus fused to ms2 polymerase were expressed in e. coli. the synthesis rate varied between 3 and 25% of the total cellular proteins. blv-ms2 polymerase fusion proteins were detected immunologically using rabbit anti-blv sera, monoclonal antibodies and a serum of a blv-infected cow.	1991	1686964
[antibody response in mice, rabbits and pigs in response to vaccination with an inactivated oil vaccine containing rotavirus and escherichia coli strains with k88, k99 and 987p fimbrial antigens].	in trials with mice, rabbits and weanling piglets, four experimental charges of a combined inactivated oil vaccine against diarrhoeas in mammals were tested: the vaccine was to be implanted to sows and it contained porcine rotavirus (prv); two charges also contained bovine rotavirus and bacterins of enterotoxicogenic strains of e. coli with protective antigens k88, k99 and 987p. at low starting antibody titres the twofold i.m. implantation of 0.2 ml vaccine stimulated in mice the production of a ...	1991	1686823
characterization of a new fimbrial antigen present in escherichia coli strains isolated from calves.	thirteen escherichia coli strains isolated from calves with diarrhoea, supposed to carry a common antigen were examined for their hemagglutinating activity and compared by bacterial agglutination, double diffusion in two dimensions and by crossed immunoelectrophoresis (cie). two of the strains were examined also in the electron microscope. most of the strains agglutinated red blood cells of horse, ox, guinea pig and chicken, of which the agglutination of ox erythrocytes was mannose-resistant (mr ...	1991	1686345
isolation and characterization of the adenylate cyclase structural gene of neurospora crassa.	a single gene (nac) encoding an adenylate cyclase was cloned from the genomic dna library of neurospora crassa, using the dna fragment encoding the catalytic domain of adenylate cyclase of saccharomyces cerevisiae as a probe. the open reading frame of this gene (6900 base pairs) was interrupted three time by introns. the protein encoded consists of 2300 amino acids and has adenylate cyclase activity. n. crassa adenylate cyclase has a high degree of homology with the catalytic domains of yeast an ...	1991	1680356
use of probes to detect virulence factor dna sequences in escherichia coli strains isolated from foods.	escherichia coli strains were isolated from 96 food samples (32 milks, 4 dairy products, 36 raw meats, 7 meat products, 7 sandwiches and 10 ready-to-eat meals). a total of 306 colonies was submitted to hybridization assays with dna probes for the following virulence factors: heat-labile toxins (lt-i and lt-ii), heat-stable toxins (st-h and st-p). shiga-like toxins (slt-i and slt-ii), adherence factor of enteropathogenic e. coli (eaf) and invasive factor (inv). six colonies isolated from 4 food s ...	1991	1677256
an f41-k88-related genetic determinant of bovine septicemic escherichia coli mediates expression of cs31a fimbriae and adherence to epithelial cells.	a genetic determinant related to that encoding the f41 fimbrial adhesin was cloned from a bovine septicemic isolate of escherichia coli. this determinant was found to mediate expression of morphologically distinct fimbriae in e. coli hb101. the gene encoding the fimbrial subunit protein was identified, and the nucleotide sequence was determined. homology with the amino-terminal amino acid sequence of cs31a (j. girardeau, m. der vartanian, j. ollier, and m. contrepois, infect. immun. 56:2180-2188 ...	1991	1675627
dietary selenium effects on milk eicosanoid concentration in dairy cows during coliform mastitis.	the effect of selenium deficiency on the product profile of arachidonic acid oxidation by enzymatic pathways in holstein cows with experimentally-induced coliform mastitis was investigated. the animals were fed dairy rations containing 0.05 mg se/kg dry matter, with the supplemented group receiving additional se to increase the dietary concentration to approximately 0.35 mg se/kg dry matter. cows were inoculated intracisternally with 30 colony-forming-units of escherichia coli at 14-16 weeks of ...	1991	1665569
modulation of function of bovine polymorphonuclear leukocytes and lymphocytes by high temperature in vitro and in vivo.	function of polymorphonuclear leukocytes (pmnl) and proliferation of lymphocytes after stimulation with mitogens were evaluated in vitro at incubation temperatures of 38.5 and 42 c, and after in vivo heat stress of lactating holstein cows. cytochrome-c reduction and random migration of pmnl were reduced when cells were preincubated or incubated at 42 c, but high incubation temperature had little or no effect on phagocytosis and killing of escherichia coli. proliferation of lymphocytes was reduce ...	1991	1662922
diverse proteins homologous to inositol monophosphatase.	bovine inositol monophosphatase (imp) and several homologous proteins were found to share two sequence motifs with bovine inositol polyphosphate 1-phosphatase (ipp). these motifs may correspond to binding sites within imp and ipp for inositol phosphates or for lithium, since both substances are bound by these proteins. this suggests that the proteins homologous to imp, which have diverse biological roles but whose function is not clear, may act by enhancing the synthesis or degradation of phosph ...	1991	1660408
cdna cloning and expression of bauhinia purpurea lectin.	bauhinia purpurea lectin (bpa) was purified from seeds of b. purpurea alba. the purified lectin was digested with an endoproteinase, asp-n, or trypsin and then the amino acid sequences of the resultant fragments were analyzed. furthermore, a cdna library for bpa was constructed using rna isolated from germinated bauhinia purpurea seeds. by gene cloning, the nucleotide sequence of bpa cdna and its deduced amino acid sequence were analyzed. the cloned bpa cdna comprised 1,152 nucleotides and the o ...	1991	1657898
a bovine papillomavirus e1-related protein binds specifically to bovine papillomavirus dna.	the e1 open reading frame of bovine papillomavirus (bpv) was expressed as a reca-e1 fusion protein in escherichia coli. the bacterially expressed reca-e1 protein exhibited sequence-specific dna binding activity; strong binding to the region from nucleotides 7819 to 93 on the bpv genome (designated region a) and weak binding to the adjacent region from nucleotides 7457 to 7818 (region b) were observed. the interaction between the bpv-derived reca-e1 protein and region a appeared to be highly spec ...	1991	1654443
meningitis in neonatal calves: 32 cases (1983-1990).	case records of 32 neonatal calves with the antemortem diagnosis of meningitis were reviewed. mean age at admission was 6 days (range, 11 hours to 30 days), and the most common concurrent clinical problem was diarrhea (16/32). twenty-seven of the calves were available for necropsy. at postmortem, there was evidence of septicemia in 22 (81%) of these calves. escherichia coli was the organism most frequently isolated (11/16; 69%) from the cns. the major clinical signs of cns disturbance observed o ...	1992	1644635
activation of mammalian dna ligase i through phosphorylation by casein kinase ii.	mammalian dna ligase i has been shown to be a phosphoprotein. dephosphorylation of purified dna ligase i causes inactivation, an effect dependent on the presence of the n-terminal region of the protein. expression of full-length human dna ligase i in escherichia coli yielded soluble but catalytically inactive enzyme whereas an n-terminally truncated form expressed activity. incubation of the full-length preparation from e. coli with purified casein kinase ii (ckii) resulted in phosphorylation of ...	1992	1639065
the signal for golgi retention of bovine beta 1,4-galactosyltransferase is in the transmembrane domain.	the expression and localization of bovine beta 1,4-galactosyltransferase (gal t) has been studied in mammalian cells transfected with gal t cdna constructs, and the role of the amino-terminal domains of gal t in golgi localization examined. here we demonstrate that the transmembrane (signal/anchor) domain of bovine gal t contains a positive golgi retention signal. bovine gal t was characterized in transfected cells with anti-bovine gal t antibodies, affinity-purified from a rabbit antiserum usin ...	1992	1637374
functional interactions of stimulatory and inhibitory gdp/gtp exchange proteins and their common substrate small gtp-binding protein.	smg gds and rho gdi are stimulatory and inhibitory gdp/gtp exchange proteins, respectively, for a group of ras p21-related small gtp-binding proteins (g proteins). rho p21 is a common substrate small g protein for both gdp/gtp exchange proteins. we examined here the functional interactions of these gdp/gtp exchange proteins with rho p21 as a substrate. smg gds and rho gdi interacted with the gdp-bound form of rho p21 and thereby stimulated and inhibited, respectively, the dissociation of gdp. th ...	1992	1634508
plasmid content and localisation of the stai (stap) gene in enterotoxigenic escherichia coli with a non-radioactive polynucleotide gene probe.	enterotoxigenic escherichia coli (etec) strain p2200 of porcine origin possessed eight possibly plasmid-determined characters (k88+ raf+ hly+ col+ smr tcr su(r) sta+) and six plasmid dna bands of 4.2-93 kb. analysis of the spontaneous loss of characters and the results of matings with other e. coli strains revealed that the k88, raf, hly, smr, tcr and su(r) characters could be transferred, and that the presence of the k88 and raf characters was associated with an 83-kb plasmid. the presence and ...	1992	1629900
the effect of cyclosporin a on cationized bovine serum albumin-induced nephropathy in nzw rabbits.	the effect of cyclosporin a (cya) was studied on the morphology and protein excretion of a rabbit chronic serum sickness nephritis using cationized bovine serum albumin (cbsa). one group of rabbits was given intravenous (i.v.) immunizing doses of cbsa and escherichia coli endotoxin. one week later, these animals began a 6-week i.v. injection schedule of cbsa only. a second group followed the same injection protocol, but was given intramuscular (i.m.) cya for 3 days prior to the immunizing dose o ...	1992	1625057
dipstick immunoassay to detect enterohemorrhagic escherichia coli o157:h7 in retail ground beef.	a sensitive and easy-to-perform dipstick immunoassay to detect escherichia coli o157:h7 in retail ground beef was developed by using a sandwich-type assay (with a polyclonal antibody to e. coli o157 as the capture antibody and a monoclonal antibody to e. coli o157:h7 as the detection antibody) on a hydrophobic polyvinylidine difluoride-based membrane. e. coli o157:h7 in ground beef could be detected within 16 h, including incubation for 12 h in enrichment broth and the immunoassay, which takes 4 ...	1992	1622249
preparation, purification, and determination of the biological activities of 12 n terminus-truncated recombinant analogues of bovine placental lactogen.	removal of 13 to 15 amino acids from the n terminus of bovine placental lactogen (bpl), which according to the three-dimensional structure of pgh corresponds to a nonhelical part of bpl, did not effect its secondary structure or change the monomer content of the protein preparation. however, it remarkably decreased the binding of the prolactin (prl) type of receptors on nb2 cells with subsequent reduction in bioactivity. the binding to the growth hormone (somatogen) receptors either did not chan ...	1992	1618771
characterization of bovine bladder mucin fractions that inhibit escherichia coli adherence to the mucin deficient rabbit bladder.	we have previously shown that a dialyzed, lyophilized saline extract from bovine urothelium can restore the antiadherence activity of the rabbit bladder following mucin removal with 50% acetone. this report describes results of initial purification of antiadherence factor(s) from bovine bladder mucin and describes results of biochemical analysis in an attempt to elucidate possible mechanism of this antiadherence activity. separation performed by gel filtration (spectra gel aca 34, 20-350 kd rang ...	1992	1613864
the relation between the soluble factor associated with h(+)-transhydrogenase of rhodospirillum rubrum and the enzyme from mitochondria and escherichia coli.	although in mitochondria, escherichia coli and rhodobacter capsulatus the h(+)-transhydrogenases are intrinsic membrane proteins, in rhodospirillum rubrum a water-soluble component (ths) and a membrane-bound component are together required for activity. ths was selectively removed from chromatophore membranes of rhs. rubrum and was purified to homogeneity by precipitation with (nh4)2so4 and ion-exchange, affinity dye and gel exclusion chromatography. the latter indicated an mr of approx. 74,000 ...	1992	1610876
distribution and characteristics of verocytotoxigenic escherichia coli isolated from ontario dairy cattle.	faecal swabs obtained from a random sample of 1131 cows and 659 calves on 100 southern ontario dairy farms were examined for verocytotoxigenic escherichia coli (vtec) using a vero cell assay. five isolates from each positive culture were tested similarly. positive colonies were examined with dna probes for shiga-like toxin i (slt-i) and slt-ii sequences. probe-negative colonies were tested for neutralization of verocytotoxicity using anti-slt-i and anti-slt-iiv antisera. colonies showing no neut ...	1992	1601076
identification of the bactericidal domain of lactoferrin.	we report the existence of a previously unknown antimicrobial domain near the n-terminus of lactoferrin in a region distinct from its iron-binding sites. a single active peptide representing this domain was isolated following gastric pepsin cleavage of human lactoferrin, and bovine lactoferrin, and sequenced by automated edman degradation. the antimicrobial sequence was found to consist mainly of a loop of 18 amino acid residues formed by a disulfide bond between cysteine residues 20 and 37 of h ...	1992	1599934
intramammary administration of gentamicin as treatment for experimentally induced escherichia coli mastitis in cows.	in 8 holstein cows, 50 colony-forming units (cfu) of escherichia coli was administered into 1 mammary gland. infections were established in all inoculated glands. in 4 of the 8 cows, 500 mg of gentamicin sulfate was administered by intramammary infusion 14 hours after inoculation; the other 4 cows were untreated controls. infusions of gentamicin also were given after each of the 3 successive milkings after the initial infusion, so that a total dose of 2 g of gentamicin was given to each of the t ...	1992	1595964
adaptation of two commercially available dna probes for the detection of e. coli and staphylococcus aureus to selected fields of dairy hygiene--an exemplary study.	the application of two commercially available colorimetric dna hybridization tests (gene-trak e. coli and staphylococcus aureus) to selected aspects of dairy hygiene was investigated. bacterial isolates of different origin, naturally contaminated cheese varieties, nonfat dry milk, milk concentrates, artificially contaminated milk and raw milks from udder quarters were examined. based on the observation that the sensitivity of the e. coli dna probe was comparable to that of the beta-d-glucuronida ...	1992	1575881
immune response of cattle to haemophilus somnus lipid a-protein conjugate vaccine and efficacy in a mouse abortion model.	immunogenicity of the lipid a component of haemophilus somnus lipooligosaccharide in cattle and mice was examined after purification, detoxification, and covalent conjugation to a protein carrier. after 2 inoculations, a substantial antibody response was induced in most cattle to lipid a and the protein carrier. to determine whether antibodies to lipid a would be protective, 5 x 10(7) colony-forming units of h somnus strain 649 were administered iv to endotoxin-responsive (c3h/hen) mice. in one ...	1992	1575380
a cloned dna probe for cowdria ruminantium hybridizes with eight heartwater strains and detects infected sheep.	the dna probe pcs20, which was cloned from the dna of the crystal springs heartwater strain from zimbabwe, cross-reacted with dnas of heartwater strains from all endemic areas, including four heartwater strains from zimbabwe, two strains from south africa, one strain from nigeria, and the gardel strain from the caribbean island of guadeloupe. by nucleic acid hybridization, the pcs20 dna probe detected cowdria ruminantium dna in all dna preparations made from plasma samples from infected sheep be ...	1992	1572987
high cortisol concentrations and mediation of the hypogalactia during endotoxin-induced mastitis.	a series of experiments was conducted to define the role of cortisol in the hypogalactia during endotoxin-induced mastitis. in the first experiment, three of six nonmastitic cows were given a continuous infusion of trilostane, a 3 beta-hydroxysteroid dehydrogenase inhibitor that blocks enhanced cortisol synthesis. trilostane had no effect in these cows. in the second experiment, six midlactation cows were given 10 micrograms of endotoxin in each of two homolateral quarters to induce mastitis. th ...	1992	1569265
enhancement of the immune response by intradermal vaccination in cattle with enterotoxigenic escherichia coli (etec) vaccine without adjuvant.	a comparison was made of the efficiency of intradermal and subcutaneous routes of vaccination in immunizing cattle with a killed vaccine of escherichia coli k99+. in this study 135 heifers aged 8-18 months were included. the effect of the number of intradermal injection sites was examined, as well as the optimal time interval between the initial and booster intradermal vaccination. the intradermal route was found to be significantly superior to the subcutaneous route. the optimal time for intrad ...	1992	1561828
the economic implications of bioengineered mastitis control.	this paper estimates the cost of mastitis for the new york dairy sector. the average cost is found to be $125 per cow from reduced milk production, treatment, and increased culling. at the 1988 cow inventory, this translates to approximately $100 million annually for the entire dairy farm sector. when quality and production losses for the processing sector are added, the cost to the new york industry alone is nearly $150 million annually. two promising new treatments, a bacteriocin and a vaccine ...	1992	1560148
cell selection strategies for making antibodies from variable gene libraries: trapping the memory pool.	the b cells of immunized animals can be used as a source of variable region (v) gene libraries. such libraries offer a way of making antibodies directly in bacteria: rearranged v genes are amplified using the polymerase chain reaction, cloned and expressed as soluble fragments in bacteria, and then screened for antigen binding. here we have used a model system to investigate antigen-selected b cells as a source of v gene libraries. mice were immunized with (4-hydroxyl-3-nitrophenyl)acetyl (np)-c ...	1992	1547829
field trial to determine efficacy of an escherichia coli j5 mastitis vaccine.	efficacy of an escherichia coli (o111:b4) j5 bacterin for preventing naturally occurring imi and clinical mastitis was tested in a 2.5-yr field trial in a 225-cow commercial herd. cows with odd-numbered identification were vaccinated, and cows with even-numbered identification served as unvaccinated controls for each lactation during the study. immunizations were subcutaneous on the upper part of the rib cage just posterior to the scapula at drying off, 30 d after drying off, and at calving. per ...	1992	1541745
opsonic activity of bovine serum and mammary secretion after escherichia coli j5 vaccination.	six pairs of cows were used to determine the effects of immunization with an escherichia coli (o111:b4) j5 bacterin on in vitro opsonization of a smooth heterologous strain of e. coli. one cow in each pair was either immunized with the vaccine or sham-immunized at drying off, 30 d after drying off, and at calving. opsonizing bacteria with serum collected from vaccinated cows 21 d after calving resulted in higher mean number of intracellular bacteria per phagocytosing neutrophil than opsonizing b ...	1992	1541744
high-level expression of functional human cytochrome p450 1a2 in escherichia coli.	enzymatically active human cytochrome p450 1a2 was expressed in escherichia coli utilizing the pcwori+ vector containing a modified cdna. the coding sequence for the nh2-terminal region of the protein was modified by the alignment and substitution of a 27 bp segment from a modified bovine p450 17a1 cdna onto the 5' end of the open reading frame of p450 1a2 at amino acid 21. the expressed chimeric p450 was produced at a high level in a functionally intact form, as assayed by the formation in vivo ...	1992	1537466
the antibacterial peptide seminal plasmin alters permeability of the inner membrane of e. coli.	seminal plasmin (spln) a 47-residue peptide, isolated from bovine seminal plasma, exhibits antibacterial activity against gram-positive and gram-negative bacteria. although spln strongly inhibits the transcription of various natural and synthetic templates by e. coli rna polymerase in vitro, it also associates with model membranes of phosphatidylcholine and phosphatidic acid. we have undertaken experiments to ascertain whether spln permeabilizes the bacterial inner membrane and thereby exerts it ...	1992	1535050
molecular cloning of a cdna encoding chicken t-protein of the glycine cleavage system and expression of the functional protein in escherichia coli. effect of mrna secondary structure in the translational initiation region on expression.	dna clones encoding chicken t-protein of the glycine cleavage system were isolated from chicken liver lambda gt10 cdna libraries. three overlapping clones provided an open reading frame of 1176 nucleotides that predicts a polypeptide of 392 amino acids (m(r) 42,056) comprised of a 16-residue mitochondrial targeting sequence and a 376-residue mature protein (m(r) 40,292). the amino acid sequence predicted for the mature protein showed 67% identity with that of bovine t-protein. a cdna encoding ma ...	1992	1526969
function of the post-translationally modified c-terminal region of rho p21.	rhoa p21, a ras p21-like small gtp-binding protein, purified from bovine aortic smooth muscle is similarly modified at its c-terminal region as described for ras p21s. in this study, i examined the role of the post-translational modifications of the c-terminal region of rhoa p21 by comparing bovine rhoa p21 with bacterially-produced rhoa p21 because the bacterial protein was not modified. bovine rhoa p21 bound to plasma membranes, but bacterial rhoa p21 did not. both the stimulatory and inhibito ...	1992	1518273
three-dimensional structure in solution of acyl-coenzyme a binding protein from bovine liver.	the three-dimensional structure of acyl-coenzyme a binding protein as encoded by the recombinant gene in escherichia coli has been determined using nuclear magnetic resonance (n.m.r.) spectroscopy. the structure consists of four alpha-helices a1 (residues 3 to 15), a2 (residues 20 to 36), a3 (residues 51 to 60), and a4 (residues 65 to 85). a1 and a4, and a2 and a3, run in parallel pairs. a2 runs anti-parallel to a1 and a4. the three-dimensional structure of the protein is reminiscent of a shallo ...	1992	1518047
determination of the carbohydrate-binding site of bauhinia purpurea lectin by affinity chromatography.	to determine the carbohydrate-binding site of bauhinia purpurea lectin (bpa), a d-galactose- and lactose-binding lectin, a peptide which interacts with lactose was purified from endoproteinase asp-n digests of bpa by chromatography on a lactose-sepharose column. it consists of nine amino acids and its amino acid sequence is asp-thr-trp-pro-asn-thr-glu-trp-ser. a tryptic fragment with the ability to interact with lactose was also purified and found to contain this sequence, consisting of nine ami ...	1992	1517321
fate of escherichia coli o157:h7 as affected by ph or sodium chloride and in fermented, dry sausage.	the influence of ph adjusted with lactic acid or hcl or sodium chloride concentration on survival or growth of escherichia coli o157:h7 in trypticase soy broth (tsb) was determined. studies also determined the fate of e. coli o157:h7 during the production and storage of fermented, dry sausage. the organism grew in tsb containing less than or equal to 6.5% nacl or at a ph of 4.5 to 9.0, adjusted with hcl. when tsb was acidified with lactic acid, the organism grew at ph 4.6 but not at ph 4.5. a co ...	1992	1514799
polymerase chain reaction for detection of verocytotoxigenic escherichia coli isolated from animal and food sources.	animals and their by-products have been implicated as important sources of verocytotoxigenic escherichia coli (vtec) associated with disease in humans. vtec comprise a wide range of serotypes and produce a variety of closely related verocytotoxins (vt). a pair of oligonucleotide primers, targeting conserved sequences found in vt1, vt2 and vte genes, was used to develop a polymerase chain reaction (pcr) procedure to detect all types of vtec. supernatants of boiled broth cultures of vtec (223 stra ...	1992	1513344
small acidic peptides from wheat germ chromatin. i. isolation and biochemical characterization.	rna synthesis in cell and cell-free systems is inhibited by a family of acidic, low molecular weight chromatin peptides (cps). these peptides were extracted from deproteinized dna of prokaryotic and eukaryotic cells, but the low yield of purified material by this procedure hinders efforts aimed at understanding their action mechanism in gene regulation. in this report we describe two purification methods of cps from an easily available source, wheat germ. a comparison is made between the method ...	1992	1508994
two linked genes for outer membrane proteins are absent in four non-disease strains of haemophilus somnus.	linked genes encoding two outer membrane proteins (p76 and a family of proteins, p120) of the bovine pathogen, haemophilus somnus, were investigated. the p120 group was previously shown to have immunoglobulin-binding activity and to react with polyclonal antiserum specific for a 270 kda antigen (p270) which also had immunoglobulin fc-binding activity. by western blotting we showed that the p76 antigen also reacted with this antiserum. the p270, p120, and p76 antigens were undetectable in four se ...	1992	1508038
the spectrum of antibiotic resistance in human and veterinary isolates of escherichia coli collected from 1984-86 in northern india.	this study was undertaken to assess the spectrum of drug resistance prevalent in escherichia coli isolates from human and animal populations in northern india. three hundred and two isolates of escherichia coli isolated from various infections of humans (47 from diarrhoea; 101 from urinary tract infection) and veterinary animals (17 from poultry septicaemia; 75 from bovine diarrhoea; 14 from ovine diarrhoea and 48 from equine metritis) were studied for their susceptibility to ampicillin, cephalo ...	1992	1506331
ubiquitous presence in mammalian cells of enzymatic activity specifically cleaving 8-hydroxyguanine-containing dna.	here we report the finding of enzymatic activity that specifically cleaves dna containing 8-hydroxyguanine (oh8gua) residues in various mammalian cells. to detect this activity, we used a synthetic double-stranded dna containing a single oh8gua at a defined position as the substrate, and analyzed the products of enzymatic digestion by polyacrylamide gel electrophoresis. two cleavage sites near the oh8gua residue were detected with partially purified fractions from cow brain and rat liver, and al ...	1992	1506269
adherence and colonization by bacterial pathogens in explant cultures of bovine mammary tissue.	explant cultures of bovine mammary tissue taken from virgin heifers were used to examine adherence, colonization and cytopathogenesis of streptococcus uberis, streptococcus agalactiae, streptococcus dysgalactiae, staphylococcus aureus and escherichia coli in the putative target tissue. none of the five bacteria was able to adhere to healthy ductular epithelium but all showed a marked tropism for exposed connective tissue. s. aureus and e. coli induced a marked cytopathic effect in ductular epith ...	1992	1496817
a specific dna probe which identifies babesia bovis in whole blood.	a genomic library of babesia bovis dna from the mexican strain m was constructed in plasmid pun121 and cloned in escherichia coli. several recombinants which hybridized strongly to radioactively labeled b. bovis genomic dna in an in situ screening were selected and further analyzed for those which specifically hybridized to b. bovis dna. it was found that pmu-b1 had the highest sensitivity, detecting 25 pg of purified b. bovis dna, and 300 parasites in 10 microliters of whole infected blood, or ...	1992	1496779
fever and acute phase response induced in dwarf goats by endotoxin and bovine and human recombinant tumour necrosis factor alpha.	tumour necrosis factor (tnf), a polypeptide produced by mononuclear phagocytes, has been implicated as an important mediator of inflammatory processes and of clinical manifestations in acute infectious diseases. to study further the potential role of tnf in infectious diseases, recombinant escherichia coli (e. coli) derived human (r.hutnf-alpha) and bovine tnf (r.botnf-alpha) were intravenously (i.v.) administered in dwarf goats. rectal temperature, heart rate, rumen motility, plasma zinc and ir ...	1992	1487832
glutathione reductases from a variety of sources are inhibited by physiological levels of glutathione.	1. glutathione reductase from human platelets, bovine intestinal mucosa, yeast and e. coli were inhibited in vitro by physiological levels of reduced glutathione with ic50s of 6.61 mm, 2.92 mm, 2.40 mm and 12.11 mm, respectively. 2. a steady-state kinetic examination revealed that glutathione inhibited the nadph oxidation (at constant [glutathione-disulphide]) catalysed by the eucaryotic enzymes uncompetitively, whereas the e. coli enzyme appeared unaffected by glutathione concentrations of up t ...	1992	1478068
rapid and sensitive method for detection of shiga-like toxin-producing escherichia coli in ground beef using the polymerase chain reaction.	a rapid and sensitive method for detection of shiga-like toxin (slt)-producing escherichia coli (slt-ec) with the polymerase chain reaction (pcr) is described. two pairs of oligonucleotide primers homologous to slti and sltii genes, respectively, were used in multiplex pcr assays. the first pair generated a ca. 600-bp pcr product with dna from all slti-producing e. coli tested but not from e. coli strains that produce sltii or variants of sltii. the second pair generated a ca. 800-bp pcr product ...	1992	1476425
susceptibility of escherichia coli isolated from intramammary infections to phagocytosis by bovine neutrophils.	thirteen escherichia coli isolated from naturally occurring imi were tested for susceptibility to phagocytosis by bovine blood neutrophils. isolates were opsonized in pooled serum collected from nine healthy lactating cows. bacteria isolated from imi first diagnosed within 3 d after calving were more resistant to phagocytosis than were isolates from imi originating during either the first half of the dry period or later during lactation. duration of the imi was negatively correlated with both ph ...	1992	1474201
[virulence factors in strains of escherichia coli isolated in urinary tract infections].	in 21 strains of e. coli isolated from patients with cystitis and asymptomatic bacteriuria the authors assessed the group, mannososensitive and mannoresistant agglutination of human, bovine, chick, guinea pig, sheep and pig erythrocytes, the production of haemolysis, colicins, aerobactin, the capacity of strains to induce oedema on mouse paws, the lethal effect in mice, the resistance to tetracycline, streptomycin, chloramphenicol, kanamycin, ampicillin and sulfamethoxidine, the transmission of ...	1992	1464082
bacterial expression, purification, and in vitro n-myristoylation of hiv-1 p17gag.	the coding region of the n-terminal 17-kda portion of hiv-1 pr55gag (p17gag) was cloned into the pet-3c expression vector and was used to overexpress hiv-1 p17gag in escherichia coli. induction of the transformed bacteria caused the accumulation of a 17-kda polypeptide in the soluble cell fraction which was released by sonication in hypotonic nondetergent buffer. the 17-kda polypeptide was purified by ammonium sulfate precipitation and successive chromatography on g-75 sephadex, deae-sephacel, a ...	1992	1458053
protein engineering of the milk-clotting aspartic proteinases.	calf chymosin and a fungal protease from mucor pusillus (mucor rennin) are members of the aspartic proteinases used as milk-coagulants in cheese industry. a system for production of recombinant chymosin as inclusion bodies in escherichia coli cells and its refolding into the active form was established. another expression system for production of mucor rennin in saccharomyces cerevisiae was also established. mucor rennin was efficiently excreted from the yeast host as a heavily glycosylated form ...	1992	1455180
genetic control of resistance to enterotoxigenic escherichia coli in infant mice.	dba/2 and cba infant mice orally challenged with bovine enterotoxigenic escherichia coli (etec) strain b80 presented resistance and susceptibility respectively, as measured by mortality rates 6 days after inoculation. serum antibodies agglutinating etec strain b80 had very low titers in both mouse strains. mendelian analysis of resistance on f1 and on segregating back-crosses showed that resistance is genetic and dominant. dominance may be explained either by a mixed control with an overdominant ...	1992	1453928
ovine pulmonary surfactant induces killing of pasteurella haemolytica, escherichia coli, and klebsiella pneumoniae by normal serum.	pulmonary surfactant has been shown to play an increasingly important role in bacterial clearance at the alveolar surface in the lung. this study describes a bactericidal mechanism in which ovine pulmonary surfactant induces killing of pasteurella haemolytica by normal serum. to demonstrate killing, six bacterial species were incubated first with pulmonary surfactant for 60 min at 37 degrees c and then with serum for an additional 60 min at 37 degrees c. p. haemolytica type a1 strains 82-25 and ...	1992	1452351
demonstration of similar calcium dependencies by mammalian high and low molecular mass phospholipase a2.	the in vitro ca2+ dependencies of arachidonyl (aa)-selective high molecular mass phospholipase a2 (hmm, 85 kda-pla2) and human low molecular mass (lmm-type ii, 14 kda)-pla2 were compared. when the lmm-pla2 and hmm-pla2 enzymes were examined for hydrolysis against [3h]aa escherichia coli in an ethyleneglycol-bis(beta-aminoethyl ether) n,n,n',n'-tetraacetic acid (egta)-free buffer system, neither enzyme demonstrated activity below 10 microm free ca2+. beyond 11 microm ca2+ both enzyme activities i ...	1992	1449538
high-level expression in escherichia coli of enzymatically active fusion proteins containing the domains of mammalian cytochromes p450 and nadph-p450 reductase flavoprotein.	this report describes the properties of two mammalian cytochromes p450 that have been expressed at high levels in escherichia coli as enzymatically active fusion proteins containing the flavoprotein domain of rat nadph-cytochrome p450 reductase (ec 1.6.2.4). fusion proteins were prepared by engineering the cdnas for the steroid-metabolizing bovine adrenal p450 17a with the cdna for rat liver nadph-p450 reductase with the introduction of a ser-thr linker to give a protein we have named rf450[mbov ...	1992	1438282
identification and expression of a fasciola hepatica gene encoding a gut antigen protein bearing repetitive sequences.	a fasciola hepatica cdna clone of about 2 kb was isolated from an expression library by immunological screening using blood serum from an experimentally infected calf. the cdna clone hybridised to a rna of about 3 kb in a northern blot experiment. the nucleotide sequence of the cdna revealed the presence of an open reading frame of 1636 bp which encoded 24 tandemly arranged 20-amino acid-long repeats, followed by 65 non-repeated residues preceding the stop codon. this antigen was expressed in es ...	1992	1435867
[isolation of vero-cytotoxin-producing escherichia coli from cattle and serotyping and toxin-typing of the isolated strains].	two hundred and sixty-six piglets with diarrhea (from 4 farms), 73 healthy pregnant pig (from 2 farms), 27 calves with diarrhea (from 9 farms) and 47 healthy milk cows (from 1 farm) were examined for vero-cytotoxin-producing escherichia coli (vtec), and 52, 11, 15 and 67 strains of vtec were isolated from 17 piglets, 11 pregnant pigs, 6 calves and 23 milk cows, respectively. all vtec strains from the piglets produced only vt2vp, while the strains from the healthy pigs did not produce vt2vp, but ...	1992	1431372
effect of intrauterine bacterial infusions and subsequent endometritis on prostaglandin f2 alpha metabolite concentrations in postpartum beef cows.	multiparous angus and crossbred angus cows were used to determine the effect of induced endometritis on plasma concentrations of 13,14-dihydro-15-keto-prostaglandin f2 alpha (pgfm) and progesterone (p4) and on duration of the estrous cycle of treatment. beginning on the day of calving (d 0), blood samples were collected on alternate days. on three consecutive days, ranging from d 8 to 14 of the first postpartum estrous cycle, uterine horns were inoculated transcervically with either 3 x 10(9) co ...	1992	1429292
[p26--a calcium binding protein from photoreceptor cells in the bovine retina: primary structure and expression in e. coli].	the primary structure of the bovine retinal calcium binding protein p26 has been determined by the parallel analysis of the protein and the corresponding cdna. this protein is identical to recovering and shares 59% homology with visinin, a cone specific calcium binding protein from chicken retina. p26 was expressed in e. coli as a fusion protein and, after purification by affinity chromatography on igg-sepharose 6, cleaved off with enteropeptidase.	1992	1417990
antibacterial effect of bovine milk antibody against escherichia coli in a mouse indigenous infection model.	a skim-milk fraction and a whey-protein concentrate (wpc) fraction were prepared from the cows that had been immunized with e. coli isolated from the mouse intestine. the antibacterial effect of these fractions against e. coli was examined. they contained antibody with a high affinity for e. coli strain 48, a representative strain in the mouse intestine, which is composed of a large amount of igg and smaller amounts of iga and igm. although these fractions showed no bactericidal or bacteriostati ...	1992	1406460
effect of interferon-gamma on the production of tumor necrosis factor during acute escherichia coli mastitis.	the effects of recombinant interferon-gamma on the production of tumor necrosis factor in 10 dairy cows with escherichia coli mastitis were determined. prophylactic administration of recombinant interferon-gamma prior to the experimental e. coli challenge was effective in modifying the production of endogenous tumor necrosis factor during acute stages of disease. elevated tumor necrosis factor concentrations were especially evident in cows that developed severe clinical symptoms and eventually d ...	1992	1401365
the heat-shock protein clpb in escherichia coli is a protein-activated atpase.	the clpb gene in escherichia coli encodes a heat-shock protein that is a close homolog of the clpa gene product. the latter is the atpase subunit of the multimeric atp-dependent protease ti (clp) in e. coli, which also contains the 21-kda proteolytic subunit (clpp). the clpb gene product has been purified to near homogeneity by deae-sepharose and heparin-agarose column chromatographies. the purified clpb consists of a major 93-kda protein and a minor 79-kda polypeptide as analyzed by polyacrylam ...	1992	1400361
serogroups of escherichia coli strains producing cytotoxic necrotizing factors cnf1 and cnf2.	the serogroups of 396 necrotizing escherichia coli of human and bovine origin isolated in spain between 1979 and 1991 have been determined. the 270 cytotoxic necrotizing factor strains belonged to 22 different o serogroups; however, 84% (226 of 270) were of one of seven serogroups (o2, o4, o6, o14, o22, o75 and o83). although necrotizing e. coli producing cytotoxic necrotizing factor 2 belonged to 28 different serogroups, only six of them (o1, o3, o15, o55, o88 and o123) accounted for 60% (76 of ...	1992	1398031
mapping of sequential epitopes recognized by monoclonal antibodies on the bovine leukaemia virus external glycoproteins expressed in escherichia coli by means of antipeptide antibodies.	a lambda gt11 cdna library prepared from bovine leukaemia virus (blv)-producing ovine cells was screened with a cocktail of anti-blv gp51 monoclonal antibodies (mabs). four recombinant phages with inserts of about 2-5 kbp were isolated. one, lambda blv-gp51-1, was sequenced and shown to encode the c-terminal part of gp51 and all of gp30. this insert was subcloned into pev-vrf1 and expressed in escherichia coli n-4830-1 cells. the blv product and a series of antipeptide antibodies were used to lo ...	1992	1383413
mip protein of legionella pneumophila exhibits peptidyl-prolyl-cis/trans isomerase (pplase) activity.	legionella pneumophila is an intracellular parasite which is able to survive and multiply in human monocytes and alveolar macrophages. the mip (macrophage infectivity potentiator) protein has been shown to be an essential virulence factor. a search of translated nucleic acid data bases has shown that the mip protein from strain wadsworth possesses regions homologous to those found in the fk506-binding proteins (fkbps) of several different eukaryotic organisms. fkbps are able to bind to the immun ...	1992	1379319
molecular cloning, stage-specific expression and cellular distribution of a putative protein kinase from plasmodium falciparum.	a putative protein kinase gene (pfpk2) has been isolated from the human parasite plasmodium falciparum by using a mixed oligonucleotide pool which corresponds to a highly conserved region of serine/threonine protein kinases. the complete nucleotide sequence of 5 kb suggests the existence of a second transcriptional unit besides that of the pfpk2 gene, separated by a highly (a+t)-rich region and transcribed in a different orientation. no intron sequence exists in pfpk2. the predicted amino acid s ...	1992	1378403
the sequence of the flavoprotein subunit of bovine heart succinate dehydrogenase.	the cdna sequence of the flavoprotein subunit of bovine heart succinate dehydrogenase is reported. this is the first complete eukaryotic sequence of the flavoprotein subunit to be characterized, and it encodes a 665-amino acid protein that consists of a presequence and a 621-residue mature protein. the deduced bovine sequence shows homology to the corresponding peptides of prokaryotic succinate dehydrogenase and the related fumarate reductases; in particular, there is good overall homology (48%) ...	1992	1375942
molecular and immunological characterization of adp-ribosylarginine hydrolases.	mono-adp-ribosylation is a reversible modification of proteins with nad:arginine adp-ribosyltransferases and adp-ribosylarginine hydrolases catalyzing the forward and reverse reactions, respectively. hydrolase activities were present in a variety of animal species, with the highest specific activities found in rat and mouse brain, spleen, and testis. rat and mouse hydrolases were dithiothreitol- and mg(2+)-dependent, whereas the bovine and guinea pig enzymes were dithiothreitol-independent. a ra ...	1992	1375222
the stability and functional properties of proteoliposomes mixed with dextran derivatives bearing hydrophobic anchor groups.	liposomes composed of escherichia coli phospholipid were coated with polysaccharides bearing hydrophobic palmitoyl anchors. the effect on the stability of liposomes without or with integral membrane proteins was investigated. a high concentration of hydrophobized dextrans protected the liposomes against detergent degradation, decreased the fluidity of the membranes, prevented fusion of the liposomes and enhanced their stability. proteoliposomes containing beef heart cytochrome-c oxidase and the ...	1992	1374644
renaturation of citrate synthase: influence of denaturant and folding assistants.	citrate synthase (cs), which has been denatured in either guanidine hydrochloride (gdnhcl) or urea can be assisted in its renaturation in a variety of ways. the addition of each of the assistants--bovine serum albumin (bsa), oxaloacetate (oaa), and glycerol--promotes renaturation. in combination, the effect of these substances is additive with respect to the yield of folded cs. the report of buchner et al. (buchner, j., schmidt, m., fuchs, m., jaenicke, r., rudolph, r., schmid, f.x., & kiefhaber ...	1992	1363914
regulation of intestinal guanylate cyclase by the heat-stable enterotoxin of escherichia coli (sta) and protein kinase c.	the heat-stable enterotoxin of escherichia coli (sta) stimulates membrane-bound guanylate cyclase in intestinal epithelium and induces fluid and ion secretion. using the t84 human colon carcinoma cell line as a model, we observed that phorbol esters markedly enhanced sta-stimulated cyclic gmp accumulation in t84 cells (c. s. weikel, c. l. spann, c. p. chambers, j. k. crane, j. linden, and e. l. hewlett, infect. immun. 58:1402-1407, 1990). in this study we document that the phorbol ester treatmen ...	1992	1360449
purification and characterization of the blue-green rat phaeochromocytoma (pc12) tyrosine hydroxylase with a dopamine-fe(iii) complex. reversal of the endogenous feedback inhibition by phosphorylation of serine-40.	tyrosine hydroxylase (th) was purified from tumours of rat phaeochromocytoma (pc12) cells by a three-step purification procedure giving 30 mg of pure enzyme in 3 days. the enzyme sedimented with an s(eo),w value of 9.2 s and revealed an apparent subunit molecular mass of 62 kda with a minor 60 kda component. two-dimensional gel isoelectric focusing/electrophoresis and tryptic digestion revealed that the heterogeneity could be accounted for by limited proteolysis of the 62 kda component and the p ...	1992	1352446
chaperonins groel and groes promote assembly of heterotetramers (alpha 2 beta 2) of mammalian mitochondrial branched-chain alpha-keto acid decarboxylase in escherichia coli.	we have investigated the possible role of chaperonins groel and groes in the folding and assembly of heterotetramers (alpha 2 beta 2) of mammalian mitochondrial branched-chain alpha-keto acid decarboxylase (e1) in escherichia coli. the mature e1 alpha subunit fused to maltose-binding protein (mbp) was coexpressed with mature e1 beta on the same vector in es- and el- mutant strains. only small or trace amounts of active e1 component were obtained. cotransformation of the es- mutant host with a se ...	1992	1352285
coob is required for assembly but not transport of cs1 pilin.	cs1 pili are filamentous proteinaceous appendages found on many enterotoxigenic escherichia coli (etec) strains isolated from human diarrhoeal disease. they are thought to effect colonization of the upper intestine by facilitating binding to human ileal epithelial cells. we have identified a gene, coob, which lies directly upstream of cooa, the gene that encodes the major structural cs1 protein. when translated in vitro, the protein product of coob migrates in sodium dodecyl sulphate/polyacrylam ...	1992	1348100
mammalian mitochondrial chaperonin 60 functions as a single toroidal ring.	chaperonins are thought to participate in the process of protein folding in bacteria and in eukaryotic mitochondria and chloroplasts. while some chaperonins are relatively well characterized, the structures of the mammalian chaperonins are unknown. we have expressed a mammalian mitochondrial chaperonin 60 in escherichia coli and purified the recombinant protein to homogeneity. structural and biochemical analyses of this protein establish a single toroidal structure of seven subunits, in contrast ...	1992	1346131
effects of thermostable escherichia coli enterotoxin peptide on the isolated rat kidney.	in order to compare the function of sodium transport between the intestine and renal tubule, we studied the effect of thermostable e. coli enterotoxin on rat kidneys. isolated kidneys from adult male hooded rats weighing 240-335 g were perfused with krebs-henseleit solution containing 60 mg/ml dialyzed bovine serum albumin. the effects of e. coli enterotoxin (sta; molecular weight approximately 2000; 18 amino acids with three disulfide bonds) were studied on glomerular filtration rate (gfr), net ...	1992	1342238
susceptibility to antibiotics of escherichia coli strains isolated from diarrhoeic and non-diarrhoeic livestock in trinidad.	the sensitivity of strains of escherichia coli isolated from calves, piglets, lambs and kids in trinidad to seven antibiotics was determined. two hundred and sixty-four (91.3%) of 289 strains isolated from diarrhoeic animals and 173 (87.4%) of 198 strains from non-diarrhoeic animals exhibited resistance to one or more antibiotics. the difference was not statistically significant (p > or = 0.05; x2). regardless of health status, isolates from lambs were least resistant (75.0%) and those from pigl ...	1992	1339991
molecular cloning and functional expression of cdna encoding a mammalian inorganic pyrophosphatase.	extracts of soluble proteins from bovine retina contain multiple species of inorganic pyrophosphatase (ppase) that can be resolved by hydroxylapatite or ion exchange chromatography. we have purified one of these isoforms by a combination of chromatography and electrophoresis under denaturing conditions and have partially sequenced four peptides generated from it by cnbr digestion. this sequence information was used to clone ppase cdna from a retinal cdna library. of five cdna inserts, three were ...	1992	1339444
cloning, expression, and catalytic mechanism of the low molecular weight phosphotyrosyl protein phosphatase from bovine heart.	the first representative of a group of mammalian, low molecular weight phosphotyrosyl protein phosphatases was cloned, sequenced and expressed in escherichia coli. using a 61-mer oligonucleotide probe based on the amino acid sequence of the purified enzyme, several overlapping cdna clones were isolated from a bovine heart cdna library. a full-length clone was obtained consisting of a 27-bp 5' noncoding region, an open reading frame encoding the expected 157 amino acid protein, and an extensive 3 ...	1992	1339287
cis proline mutants of ribonuclease a. i. thermal stability.	a chemically synthesized gene for ribonuclease a has been expressed in escherichia coli using a t7 expression system (studier, f.w., rosenberg, a.h., dunn, j.j., & dubendorff, j.w., 1990, methods enzymol. 185, 60-89). the expressed protein, which contains an additional n-terminal methionine residue, has physical and catalytic properties close to those of bovine ribonuclease a. the expressed protein accumulates in inclusion bodies and has scrambled disulfide bonds; the native disulfide bonds are ...	1992	1338975
characterization of the human calmodulin-like protein expressed in escherichia coli.	the protein-coding region of an intronless human calmodulin-like gene [koller, m., & strehler, e. e. (1988) febs lett. 239, 121-128] has been inserted into a pkk233-2 expression vector, and the 148-residue, m(r) = 16,800 human protein was purified to apparent homogeneity by phenyl-sepharose affinity chromatography from cultures of escherichia coli jm105 transformed with the recombinant vector. several milligrams of the purified protein were obtained from 1 l of bacterial culture. a number of pro ...	1992	1334432