Publications

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
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
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
protein phosphatase 2a is a specific protamine-kinase-inactivating phosphatase.	purified preparations of a protamine protein kinase from bovine kidney cytosol [damuni, amick & sneed (1989) j. biol. chem. 264, 6412-6416] were inactivated after incubation with near-homogeneous preparations of protein phosphatase 2a1 and protein phosphatase 2a2. these protein phosphatase 2a-mediated inactivations of the protamine kinase were unaffected by highly purified preparations of inhibitor 2, but were prevented when the incubations were performed in the presence of 100 nm microcystin-lr ...	1992	1332680
functional reconstitution of membrane proteins in monolayer liposomes from bipolar lipids of sulfolobus acidocaldarius.	membranes of sulfolobus acidocaldarius, an extreme thermophilic archaebacterium, are composed of unusual bipolar lipids. they consist of macrocyclic tetraethers with two polar heads linked by two hydrophobic c40 phytanyl chains which are thought to be arranged as a monolayer in the cytoplasmic membrane. fractionation of a total lipid-extract from s. acidocaldarius yielded a lipid fraction which forms closed and stable unilamellar liposomes in aqueous media. beef heart cytochrome c-oxidase could ...	1992	1309769
acute coliform mastitis in dairy cows: endotoxin and biochemical changes in plasma and colony-forming units in milk.		1992	1300679
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
the biological interaction of cis- and trans-urocanic acid and dna.	the potential for the cis and trans isomers of urocanic acid to produce dna damage was measured by assays for dna binding (32p-postlabeling assay), for induction of dna repair (unscheduled dna synthesis assay) and induction of mutations (salmonella typhimurium and escherichia coli plate-incorporation assays). these assays did not detect any evidence of a direct effect of either isomer of urocanic acid on dna over a wide range of concentrations. these results suggest that neither isomer of urocan ...	1992	1300140
[detection of verotoxin-producing escherichia coli using polymerase chain reaction from dairy cattle].	the vero cytotoxin (vt) is responsible for hemorrhagic colitis and hemolytic uremic syndrome. polymerase chain reaction (pcr) was used to detect vt-producing coliform bacteria from dairy cattle. it was found that 39 (33.3%) of the 117 fecal samples examined were recognized with vt genes in bglb enrichment broth by the pcr method (named bglb-pcr). of the vt-positive samples, 31 samples (26.5%) were found to have vt-producing escherichia coli. frequencies of isolation in younger cattle (under 5 mo ...	1992	1293218
escherichia coli f0f1-atpase. residues involved in catalysis and coupling.	the molecular biological approach has provided important information toward understanding the complexities of the f0f1 atpase. this article focuses on our recent results on the atpase catalytic site contained in the beta subunit and the role of the gamma subunit in regulation of proton transport. we used a combination of affinity labeling and mutagenesis to locate several residues of the alpha and beta subunits in the catalytic site. adenosine triphosphopyridoxal (ap3-pl) labeled beta lys-155, b ...	1992	1288330
expression of the phosphorylase kinase gamma subunit catalytic domain in escherichia coli.	the catalytic subunit of phosphorylase b kinase (gamma) and an engineered truncated form (gamma-trc, residues 1-297) have been expressed in escherichia coli. the truncated protein included the entire catalytic domain as defined by sequence alignment with other protein kinases but lacked the putative calmodulin binding domain. full-length protein was produced in insoluble aggregates. some activity was regenerated by solubilization in urea and dilution into renaturating buffer but the activity was ...	1992	1287663
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
[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
[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
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
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
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
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
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
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
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
[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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
escherichia coli o157:h7 and its significance in foods.	escherichia coli o157:h7 was conclusively identified as a pathogen in 1982 following its association with two food-related outbreaks of an unusual gastrointestinal illness. the organism is now recognized as an important cause of foodborne disease, with outbreaks reported in the u.s.a., canada, and the united kingdom. illness is generally quite severe, and can include three different syndromes, i.e., hemorrhagic colitis, hemolytic uremic syndrome, and thrombotic thrombocytopenic purpura. most out ...	1991	1854598
the amino-terminal half of rotavirus sa114fm vp4 protein contains a hemagglutination domain and primes for neutralizing antibodies to the virus.	we have previously reported the synthesis in escherichia coli of polypeptide ms2-vp8', which contains the amino-terminal half of the sa114fm vp4 protein fused to ms2 bacteriophage polymerase sequences (c. f. arias, m. lizano, and s. lópez, j. gen. virol. 68:633-642, 1987). in this work we have synthesized the carboxy-terminal half of the vp4 protein also fused to the ms2 polymerase. this protein, designated ms2-vp5', was recognized by sera to the complete virion and was able to induce antibodies ...	1991	1847459
cryptosporidiosis in neonatal calves: 277 cases (1986-1987).	from january 1986 through december 1987, 277 cases of cryptosporidiosis in calves were diagnosed by the south dakota state university diagnostic laboratory. cryptosporidium sp was the only pathogen identified in 142 (51.3%) of the calves. concurrent infections with rotavirus, coronavirus, escherichia coli, salmonella sp, bovine viral diarrhea virus, or other pathogens were identified in the remaining 135 calves. after elimination of cases involving autolyzed specimens or calves with chronic diar ...	1991	1874678
controlling acute escherichia coli mastitis during the periparturient period with recombinant bovine interferon gamma.	the efficacy of recombinant bovine interferon (rboifn)-gamma against experimentally induced escherichia coli mastitis during the periparturient period was investigated. dairy cows intramammarily treated with rboifn-gamma 24 h before the e. coli challenge had fewer infected quarters, lower clinical scores, and infections of shorter duration when compared to placebo-treated animals. all rboifn-gamma treated cows survived the experimental e. coli challenge. however, placebo treated cows had a 42% m ...	1991	1908159
rsr1 and rap1 gtpases are activated by the same gtpase-activating protein and require threonine 65 for their activation.	the rsr1 protein of saccharomyces cerevisiae has been shown to be essential for bud site selection (bender, a., and pringle, j. (1989) proc. natl. acad. sci. u.s.a. 86, 9976-9980). this protein of 272 amino acids shares approximately 50% sequence identity with both ras and rap gtpases. however, neither gtp binding nor gtpase activity of the rsr1 protein has been reported. the rsr1 protein shares with human rap1 gtpases the four specific motifs, i.e. gly-12, residues 32-40, ala-59, and residues 6 ...	1991	1910037
interactions of the bovine brain a1-adenosine receptor with recombinant g protein alpha-subunits. selectivity for rgi alpha-3.	the ability of the bovine brain a1-adenosine receptor to discriminate between different g protein subtypes was tested using g protein alpha-subunits synthesized in escherichia coli (rg alpha-subunits). when combined with a 3-fold molar excess of beta gamma-subunit purified from bovine brain and used at high concentrations, all three subtypes of rgi alpha (rgi alpha-1, rgi alpha-2, and rgi alpha-3) and rgo alpha were capable of reconstituting guanine nucleotide-sensitive high-affinity binding of ...	1991	1917922
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
[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
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
development of genomic probes to sarcocystis cruzi (apicomplexa).	a genomic library of sarcocystis cruzi sporozoite dna was constructed in bacteriophage lambda gt10. recombinant phages containing insert dna were selected by growth on escherichia coli strain c600 hfla150. of 14 clones examined, 11 contained dna inserts ranging in size from approximately 1.45 kilobase (kb) to 6.18 kb. insert dna from four of these clones specifically hybridized to 32p-labelled s. cruzi merozoite dna. one of these insert dna, clone sl41, was selected and labelled with 32p. this p ...	1991	1837193
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
simian virus 40 vp2/3 small structural proteins harbor their own nuclear transport signal.	we have used a microinjection approach to identify a domain of the simian virus 40 (sv40) structural proteins vp2 and vp3(vp2/3) responsible for their nuclear transport. by using both synthetic peptides, containing small regions of vp2/3 conjugated to bovine serum albumin (bsa), and beta-galactosidase-vp3 fusion proteins, we have narrowed this nuclear transport signal (nts) to 9 amino acids (198 to 206 of vp3 or 316 to 324 of vp2), gly-pro-asn-lys-lys-lys-arg-lys-leu. the porter proteins carryin ...	1991	1847270
thrombin and h64a subtilisin cleavage of fusion proteins for preparation of human recombinant parathyroid hormone.	human parathyroid hormone, hpth, an 84 amino acid polypeptide, was produced intracellularly in escherichia coli as a fusion protein, linked to the c-terminus of a 15 kd igg-binding protein. approximately 100 mg fusion protein was obtained per liter fermentation medium. to test the efficiency of two alternative enzymatic cleavage methods, two fusion proteins differing only in the linker region were constructed. cleavage of a phe-phe-pro-arg linker was obtained with bovine thrombin and cleavage of ...	1991	1799410
a new diarrhoeic syndrome with ataxia in young charolais calves: clinical and microbiological studies.	a new diarrhoeic syndrome was examined clinically in 19 one to two-week old charolais calves. it differs from other digestive disorders in calves of this age in the discrete diarrhoeic signs, the absence of dehydration and the presence of signs of ataxia. the microbiological study carried out for three consecutive years in 58 sick calves and nine healthy control calves demonstrated the special role of e coli possessing virulence markers from septicaemic strains (cs31a, col v). the clinical signs ...	1991	1853535
dna hybridization and latex agglutination for detection of heat-labile- and shiga-like toxin-producing escherichia coli in meat.	dna-hybridization and latex-agglutination tests were used for screening of a group of escherichia coli isolates for heat-labile enterotoxin (lt)- and shiga-like toxin (slt1 or vt1) -producing strains, respectively. strains tested originated from 162 meat samples (poultry, pigs and beef) chosen at random. additionally lt- and slt1-producing reference strains were tested. the dna-hybridization technique allowed screening of large numbers of strains, whereas large scale testing of strains by latex ...	1991	1863526
antibodies to iron-regulated outer membrane proteins of coliform bacteria isolated from bovine intramammary infections.	expression of iron-regulated outer membrane proteins (omp) by escherichia coli and klebsiella pneumoniae initially isolated from bovine intramammary infections (imi) was investigated. additionally, the presence of antibodies in bovine serum and mammary secretion directed against the iron-regulated omp was examined. outer membrane proteins were separated by sodium-dodecyl polyacrylamide electrophoresis. detection of immunoglobulin g directed against omp was by immunoblotting. all gram-negative ba ...	1991	1866891
n-acetyl-beta-d-glucosaminidase, etiologic agent, and duration of clinical signs for sequential episodes of chronic clinical mastitis in dairy cows.	this study examined effects of repeated episodes of clinical mastitis in chronically infected quarters on milk n-acetyl-beta-d-glucosaminidase activity and duration of clinical signs. milk samples were collected at each clinical onset from 49 chronic mastitis cases on a 1700-cow michigan dairy farm. there were 49 first episodes of clinical mastitis, 49 second episodes, and 13 episodes of third or more. agents isolated were staphylococcus aureus (18.4%), staphylococcus (7.3%), no growth (20.2%), ...	1991	1880262
endometrial biopsy in holstein-friesian dairy cows. iii. bacteriological analysis and correlations with histological findings.	this study examines the results of bacterial culture from 159 endometrial biopsy samples from 97 commercial dairy cows and correlations between bacteriological and histological findings. bacteria were isolated from approximately 80% of biopsies taken at day 26 and day 40 postpartum. eleven percent of biopsies were positive for both aerobic and anaerobic culture. streptococci, escherichia coli and actinomyces pyogenes were the most common isolates. isolation of a. pyogenes from a biopsy at day 26 ...	1991	1884297
clonal relationships among escherichia coli serogroup 06 isolates from human and animal infections.	the clonal relationship of thirty e. coli strains of 0 antigen serotype 06 isolated from human, dog, pig or cow infections were investigated. two main clones with serotypes 06 : h1 or 06 : h31, h- were identified. isolates from humans, dogs, pigs and cows were found in both clones, indicating that animals are a possible source for human extraintestinal escherichia coli strains. two human etec (06 : h16) and two pig isolates (06 : h10) were not related to the 06 : h1 or 06 : h31, h- e. coli clone ...	1991	1884980
interaction of animal mitochondrial ef-tu.ef-ts with aminoacyl-trna, guanine nucleotides, and ribosomes.	the mammalian mitochondrial complex consisting of elongation factors ef-tu and ef-ts (ef-tu.tsmt) is capable of efficiently binding aminoacyl-trna to the ribosome in the presence and absence of guanine nucleotides. in the presence of gtp the binding reaction is catalytic. in the absence of guanine nucleotides, or in the presence of a non-hydrolyzable gtp analog, only one round of ribosome binding occurs. ef-tu.tsmt is capable of forming a ternary complex with gtp and escherichia coli phe-trna as ...	1991	1885567
the effect of lipopolysaccharide on bovine mammary macrophage function.	the effect of escherichia coli lipopolysaccharide (lps) on the expression of major histocompatibility complex (mhc) class ii molecules by bovine mammary macrophages was examined. the ability of lps-treated mammary macrophages to support antigen-specific t-cell proliferation, as a measure of their antigen presentation ability, was also evaluated. for this purpose, control and lps-treated macrophages were pulsed with heat-killed staphylococcus aureus and then cultured with s. aureus-sensitized t-c ...	1991	1889031
the use of plasmid profiles and nucleic acid probes in epidemiologic investigations of foodborne, diarrheal diseases.	the application of nucleic acid analyses to investigations of infectious disease outbreaks has resulted in useful molecular strain markers that distinguish the epidemic clone of a particular pathogen and help identify specific vehicles of infection. we have successfully used plasmid profile analysis, restriction endonuclease digestion of plasmid and whole-cell dnas, and nucleic acid hybridization to investigate recent outbreaks of foodborne diarrheal illness. plasmid analysis has been important ...	1991	2018708
sensitive immunoassay for rat parvalbumin: tissue distribution and developmental changes.	a sensitive enzyme immunoassay for measurements of rat parvalbumin was established using antibodies raised in rabbits with parvalbumin purified from skeletal muscles. antibodies in the antiserum were purified with a parvalbumin-coupled sepharose column. the sandwich-type immunoassay system for parvalbumin was composed of polystyrene balls with immobilized purified antibodies and the same antibodies labeled with beta-d-galactosidase from escherichia coli. the assay was highly sensitive and the mi ...	1991	1892868
study of mammalian selenocysteyl-trna synthesis with [75se]hse.	the mechanisms of the synthesis of mammalian selenocysteyl-(scy)-trna were studied using [75se]h2se. h2se was prepared from [75se]selenite, glutathione, nadph and glutathione reductase, and was purified by chromatography. it was confirmed that this h2se was a se donor in the reaction of the synthesis of scy-trna. [75se]scy, liberated from aminoacyl-trna, was analyzed by tlc on silica gel an subsequent autoradiography. the activity of scy-trna synthesis was found in the supernatant at 105,000 x g ...	1991	1894009
antigenic homology among gram-negative organisms isolated from cattle with clinical mastitis.	this study examined the degree of serologic homology among mastitis pathogens. antibodies were raised against the rc mutant, escherichia coli o111:b4 (strain j5) and affinity purified against lipopolysaccharide derived from the ra mutant, salmonella typhimurium tv119. these antibodies reacted with a battery of unrelated gram-negative bacteria in whole cell elisa. bacteria with strong cross-reactions included a heterologous, smooth e. coli, salmonella dublin, s. typhimurium, salmonella newport, a ...	1991	1907303
partial budget analysis of vaccinating dairy cattle against coliform mastitis with an escherichia coli j5 vaccine.	partial budget analysis of clinical coliform mastitis prevention supported vaccination of dairy cows with an escherichia coli j5 vaccine. increased profits of $57/cow lactation were predicted using a computer spreadsheet derived partial budget with generalized herd input data. herd vaccination programs were predicted to be profitable when greater than 1% of cow lactations resulted in clinical coliform mastitis. herd vaccination programs were predicted to be profitable at all herd milk production ...	1991	1917656
immunoreactive alpha a crystallin in rat non-lenticular tissues detected with a sensitive immunoassay method.	for the quantitative analysis of the a subunit of alpha crystallin (alpha a) in the lens and for the survey of possible existence of alpha a in the non-lenticular tissues, we have established a highly sensitive and specific immunoassay method for alpha a. antisera to alpha a were raised in rabbits with alpha a purified from bovine lens, or the c-terminal decapeptide (eekpssapss) of alpha a (alpha apep). the antibodies to alpha a and alpha apep were purified by the use of an alpha a-coupled sepha ...	1991	1932094
cdna and deduced amino acid sequence of a mouse dna repair enzyme (apex nuclease) with significant homology to escherichia coli exonuclease iii.	we purified a mouse dna repair enzyme having apurinic/apyrimidinic endonuclease, dna 3'-phosphatase, 3'-5'-exonuclease and dna 3' repair diesterase activities, and designated the enzyme as apex nuclease. a cdna clone for the enzyme was isolated from a mouse spleen cdna library using probes of degenerate oligonucleotides deduced from the n-terminal amino acid sequence of the enzyme. the complete nucleotide sequence of the cdna (1.3 kilobases) was determined. northern hybridization using this cdna ...	1991	1939131
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
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
potent bactericidal activity of bovine lactoferrin hydrolysate produced by heat treatment at acidic ph.	a hydrolysate of bovine lactoferrin produced by heat treatment under acidic conditions had antibacterial activity at concentrations of 10 micrograms/ml in the culture medium. the optimal degree of hydrolysis for this activity was about 10%. heat-treated lactoferrin, treated at ph 2.0 and 120 degrees c for 15 min and degree of hydrolysis of about 10%, had no fe-binding capacity (0%) and less antigenicity (about 10(-6) than untreated lactoferrin. heat-treated lactoferrin increased in antibacterial ...	1991	1757616
protective effect of tissue ferritins in experimental escherichia coli infection of mice in vivo.	the effect of ferritins from horse (fh) and bovine (fb) spleen and murine liver (fm) on the survival rate of cfw mice lethally infected with escherichia coli (strain 8440-78 k 80/b) was evaluated. ferritins given intravenously 24 h before intravenous inoculation of bacteria, protected mice most effectively from death due to infection. the effect was dose dependent. at 500 micrograms of ferritin per mouse, the maximum survival rates were 86% (fh), 81% (fm) and 79% (fb), while only 5% of the contr ...	1991	1768608
prevalence of antibiotic-resistant escherichia coli in danish pigs and cattle.	the present paper reports on the antibiotic resistance of e. coli isolated from danish piglets and calves in 1987-1988, and compares the results with similar investigations performed during the periods 1971-1972 and 1977-1978. rectal swabs from 52 piglets and from 78 calves were examined. all the animals studied harboured resistant e. coli. this is a significant increase compared to the previously conducted investigations. the number of strains having three or more resistance markers did not dif ...	1991	1772647
virulence factors associated with cytotoxic necrotizing factor type two in bovine diarrheic and septicemic strains of escherichia coli.	forty-three bovine isolates of escherichia coli producing a second type of cytotoxic necrotizing factor (cnf2) and three k-12 strains carrying different vir plasmids coding for cnf2 were tested for the presence of several virulence factors. most of the strains were serum resistant (79%), produced an aerobactin (70%), and adhered to calf villi (53%); some of them produced a colicin (32%) and a hemolysin (9%). these strains were also tested by a colony hybridization assay with gene probes for six ...	1991	1774259
effect of milking without pulsation on teat duct colonization with streptococcus agalactiae and penetrability to endotoxin.	forty quarters of 10 cows were milked for a 9-d period with one of four treatments: 1) no liner pulsation, 2) conventional milking, 3) no pulsation for 4 d followed by conventional milking for 5 d, and 4) conventional milking for 4 d followed by milking without pulsation for 5 d. all teat orifices were inoculated with approximately .5 million cfu of streptococcus agalactiae and streptococcus dysgalactiae on d 1 and 5. recoveries of strep. agalactiae from the teat end were increased for teats mil ...	1991	1779054
conglutinin exhibits a complement-dependent enhancement of the respiratory burst of phagocytes stimulated by e. coli.	conglutinin is a mammalian c-type lectin which shows anti-bacterial activity when tested in vivo and in vitro. this study concerns the effect of conglutinin on the respiratory burst of murine spleen cells, using a chemiluminescence assay for measurement of generated reactive oxygen metabolites. conglutinin enhances, in a dose-dependent manner, the respiratory burst of spleen cells stimulated with serum-opsonized escherichia coli. the enhancement was only demonstrable in the presence of a functio ...	1991	1783427