Publications

dietary supplementation with lactobacillus- and propionibacterium-based direct-fed microbials and prevalence of escherichia coli o157 in beef feedlot cattle and on hides at harvest.	the objective of this study was to describe the prevalence of escherichia coli o157 in the feces and on the hides of finishing beef cattle fed a standard diet and those fed diets supplemented with direct-fed microbials. two hundred forty steers received one of four treatments throughout the feeding period: (i) control: no added microbials; (ii) hnp51: high dose of lactohacillius acidophilus strain np 51 (10(9) cfu per steer daily) and propionibacterium freudenreichii (10(9) cfu per steer daily); ...	2004	15151223
purification of recombinant bovine normal prion protein prp(104-242) by hphic.	purification of the prion protein (prp) is a major concern for biological or biophysical analysis as are the structural specificities of this protein in relation to infectivity. a simple and efficient method for purification of recombinant bovine normal prion protein containing residues 104-242, prp(104-242) expressed in escherichia coli by high performance hydrophobic interaction chromatography (hphic) was presented in this work. the solution containing denatured and reduced protein in 8.0 mol/ ...	2004	15171928
phenoxazine compounds produced by the reactions with bovine hemoglobin show antimicrobial activity against non-tuberculosis mycobacteria.	we studied the anti-microbial effects of phenoxazines produced by the reaction of o-aminophenol or its derivatives with bovine hemoglobin, on seven species of mycobacteria such as mycobacterium tuberculosis, mycobacterium marinum, mycobacterium intracellulare, mycobacterium scrofulaceum, mycobacterium fortuitum, mycobacterium kansasii and mycobacterium smegmatis and some bacteria such as escherichia coli, pseudomonas aeruginosa, salmonella enterica serovar typhimurium, staphylococcus aureus, lis ...	2004	15185971
high prevalence of enterohemorrhagic escherichia coli (ehec) o157 from cattle in selected regions of japan.	the prevalence of enterohemorrhagic escherichia coli (ehec) o157 was examined in bovine faeces. ehec o157 was isolated from the faeces of 42 (13.0%) of 324 cattle. of the 4 farms and the facilities tested, the 3 farms and the facilities were found positive for ehec o157. the highest isolation rate among the farms was 33.7%. the prevalence of ehec o157 in heifers was higher than that in calves and other cattle. no cattle positive for ehec o157 showed any clinical signs except 2 calves with diarrh ...	2004	15187376
effect of recombinant cytokines on leucocytes and physiological changes in bovine mammary glands during early involution.	we examined the effects of administering recombinant bovine cytokines to non-lactating dairy cows and measured mammary gland leucocytes and the involution process. after the final milking, groups of cows were given an intramammary infusion of cytokine in two quarters. these cytokines were recombinant bovine interleukin-2 (rboll-2) (2 x 10(5) units, n = 6), recombinant bovine granulocyte-macrophage colony stimulating factor (rbogm-csf) (500 microg, n = 4) and recombinant bovine interleukin-1beta ...	2004	15190942
mild to moderate clinical mastitis: efficacy of intramammary amoxicillin, frequent milk-out, a combined intramammary amoxicillin, and frequent milk-out treatment versus no treatment.	study objectives were to determine the efficacy of 4 methods (intramammary amoxicillin, frequent milk-out, a combined intramammary amoxicillin and frequent milk-out, and no treatment) of managing mild to moderate clinical mastitis in a university dairy herd. clinical and microbiological cures, milk production, disease progression, and california mastitis tests scores were evaluated. cows meeting the study criteria were assigned to one of four treatment options based on a systematic randomization ...	2004	15202642
crystallization and preliminary x-ray diffraction study of mammalian mitochondrial seryl-trna synthetase.	the mitochondrial seryl-trna synthetase (mt serrs) from bos taurus was overexpressed in escherichia coli and crystallized using the sitting-drop vapour-diffusion method. crystals grew in a very narrow range of conditions using peg 8000 as precipitant at room temperature. an appropriate concentration of lithium sulfate was critical for crystal nucleation. crystals diffracted well beyond a resolution of 1.6 a and were found to belong to the orthorhombic space group c222(1), with unit-cell paramete ...	2004	15213404
prevalence estimation and risk factors for escherichia coli o157 on dutch dairy farms.	to estimate the prevalence of escherichia coli o157 on dutch dairy herds, faecal samples were collected once from 678 randomly selected dairy farms in the period october 1996-december 2000. samples were cultured for e. coli o157. thirty-eight isolates were tested for virulence genes (eae, vt1 and vt2). a questionnaire about farm characteristics was taken from the farm manager, resulting in variables that could be analysed to identify and quantify factors associated with presence of e. coli o157. ...	2004	15219969
effect on bovine lactoferrin on the activation of the enteroinvasive bacterial type iii secretion system.	shigella and enteroinvasive escherichia coli (eiec) strains secrete virulence proteins by a complex machinery called the type iii secretion (tts) apparatus. secretion of virulence proteins is a tightly-regulated phenomenon such that the tts system is weakly active when bacteria are grown in common laboratory media. activation of the tts system is triggered by contact with eukaryotic cells, or can be artificially stimulated by the addition of congo red dye to the growth medium. exploiting the abi ...	2004	15222475
bacterial growth during the early phase of infection determines the severity of experimental escherichia coli mastitis in dairy cows.	the aim of this study was to investigate the importance of bacterial growth for the severity of experimental escherichia coli mastitis, indirectly expressed as the area under the curve of bacterial counts in milk over time. the association of pre-infusion somatic cell count and post-infusion influx of inflammatory cells in milk with severity of infection was also examined. bacterial growth was studied through culture in milk samples (in vitro) and through monitoring of bacterial counts in milk d ...	2004	15223122
quantity and quality of runoff from a beef cattle feedlot in southern alberta.	southern alberta, which has a cold climate dominated by strong chinook winds, has the highest density of feedlot cattle in canada. however, the quantity and quality of runoff from beef cattle (bos taurus) feedlots in this unique region has not been investigated. our objectives were to compare runoff quantity (1998-2002) with catch-basin design criteria; determine concentrations of selected inorganic chemical parameters (1998-2000) in runoff in relation to water quality guidelines and the potenti ...	2004	15224948
heme a synthase does not incorporate molecular oxygen into the formyl group of heme a.	heme a is an obligatory cofactor in all eukaryotic and many prokaryotic cytochrome c oxidases. the final step in heme a biosynthesis requires the oxidation of the c8 methyl substituent on pyrrole ring d to an aldehyde, a reaction catalyzed by heme a synthase. to effect this transformation, heme a synthase is proposed to utilize a heme b cofactor, oxidizing the substrate via successive monooxygenase reactions. consistent with this hypothesis, the activity of heme a synthase is found to be strictl ...	2004	15236569
prevalence and characteristics of escherichia coli o157 from major food animals in korea.	escherichia coli o157:h7/nm (e. coli o157) is now recognized as an important cause of diarrhea, hemorrhagic colitis and hemolytic-uremic syndrome worldwide. there have been several cases of human e. coli o157 infection in korea since it was first isolated from a patient with hemolytic-uremic syndrome in 1998. meat, other foods, and recreational and drinking water contaminated with animal feces are probably the major sources of the e. coli o157 infection. in this study, we investigated the preval ...	2004	15240073
microbiological conditions of mechanically tenderized beef cuts prepared at four retail stores.	a group of 25 retail cuts of mechanically tenderized beef prepared during usual commercial operations at store facilities were obtained from each of four retail stores. aerobes, coliforms, escherichia coli and organisms that formed black or grey colonies on harlequin agar (ha), a medium formulated for the recovery of listeria, were enumerated in samples from the surfaces and the deep tissues of cuts. for the four groups of cuts, the mean numbers of aerobes on the surfaces of cuts differed by <1 ...	2004	15240079
characterization of immunodominant linear b-cell epitopes on the carboxy terminus of the rinderpest virus nucleocapsid protein.	the nucleocapsid (n) protein of rinderpest virus (rpv) is one of the most abundant and immunogenic viral proteins expressed during natural or experimental infection. to identify immunogenic epitopes on the n protein, different forms of rpv n protein, including the full-length protein (n(1-525)), an amino-terminal construct (n(1-179)), and a carboxy-terminal construct (n(414-496)), were expressed in escherichia coli as glutathione s-transferase (gst) fusion proteins. the antigenicity of each reco ...	2004	15242937
three-dimensional solution structure of the cytoplasmic b domain of the mannitol transporter iimannitol of the escherichia coli phosphotransferase system.	the solution structure of the cytoplasmic b domain of the mannitol (mtl) transporter (ii(mtl)) from the mannitol branch of the escherichia coli phosphotransferase system has been solved by multidimensional nmr spectroscopy with extensive use of residual dipolar couplings. the ordered iib(mtl) domain (residues 375-471 of ii(mtl)) consists of a four-stranded parallel beta-sheet flanked by two helices (alpha(1) and alpha(3)) on one face and helix alpha(2) on the opposite face with a characteristic ...	2004	15258141
short communication: influence of staphylococcus aureus intramammary infection on serum copper, zinc, and iron concentrations.	the goal of the present study was to characterize changes in serum trace mineral concentrations in cattle with experimentally induced staphylococcus aureus mastitis. nine primiparous holstein-friesian cattle were challenged with approximately 150 cfu of staph. aureus atcc29740 by intramammary infusion on d 6, 7, and 8 of lactation. serum cu, zn, and fe concentrations were determined immediately before and at 24, 48, and 72 h after the final intramammary infusion of staph. aureus. infection statu ...	2004	15259232
all-trans retinoic acid is increased in the acute phase-related hyporetinemia during escherichia coli mastitis.	blood vitamin a profiles, including concentrations of retinol and its active metabolite retinoic acid, were assessed during the peripartum period and during experimentally induced escherichia coli mastitis in heifers. serum retinol decreased in all animals in the immediate postpartum period and normalized within 1 wk after parturition. no significant changes were detectable in the concentrations of retinoic acid isomers during puerperium. following intramammary e. coli infusion, all cows showed ...	2004	15259233
localization of a 56-kda antigen that is present in multiple developmental stages of neospora caninum.	the purpose of the present study was to characterize the intracellular distribution of a native neospora caninum 56-kda protein that is recognized by sera from n. caninum-infected dairy cattle. the complementary dna coding for this protein was expressed in escherichia coli as a polyhis fusion protein to which antiserum was prepared and used to localize the antigen in n. caninum tachyzoites and bradyzoites. by sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoblotting, antirecomb ...	2004	15270121
persistence of enterohemorrhagic escherichia coli o157:h7 in soil and on leaf lettuce and parsley grown in fields treated with contaminated manure composts or irrigation water.	outbreaks of enterohemorrhagic escherichia coli o157:h7 infections associated with lettuce and other leaf crops have occurred with increasing frequency in recent years. contaminated manure and polluted irrigation water are probable vehicles for the pathogen in many outbreaks. in this study, the occurrence and persistence of e. coli o157:h7 in soil fertilized with contaminated poultry or bovine manure composts or treated with contaminated irrigation water and on lettuce and parsley grown on these ...	2004	15270487
microbiological quality of ground beef from conventionally-reared cattle and "raised without antibiotics" label claims.	the contamination of the food supply with pathogens and antimicrobial-resistant bacteria has emerged as an important health concern. we compared the microbiological quality of 77 samples of ground beef from conventionally raised cattle with 73 samples of ground beef from cattle raised without antimicrobial agents. contamination with coliforms (1.7 log cfu/g) and escherichia coli (0.51 log cfu/g) and shiga toxin 2-producing e. coli (6% prevalence) was similar in both sample groups. neither salmon ...	2004	15270497
campylobacter prevalence in lactating dairy cows in the united states.	the objective of the present study was to determine the prevalence of intestinal campylobacter in lactating dairy cows from various regions of the united states. participating commercial dairy farms were chosen at random and were part of a national survey to determine e. coli o157:h7 and salmonella prevalence in dairy cows. farms had no previous history of campylobacter problems. fecal samples were collected rectally from 720 cows on farms in the northeast (four farms), in the desert southwest ( ...	2004	15270504
endocrine aspects in pathogenesis of mastitis in postpartum dairy cows.	in well-managed dairy herds some environmental pathogens including gram-negative (gn) strains (e. coli and others) have been recognized recently as the predominant causative microbes of mastitis in the peri-parturient period. in early weeks of lactation hyperketonaemia may predispose the high-producing cows for gn mastitis. in gn mastitis cytokines, eicosanoids and oxygen radicals are released, which are responsible for the local and systemic symptoms. experimental administration of endotoxin in ...	2004	15271468
direct binding of visual arrestin to microtubules determines the differential subcellular localization of its splice variants in rod photoreceptors.	proper function of visual arrestin is indispensable for rapid signal shut-off in rod photoreceptors. dramatic light-dependent changes in its subcellular localization are believed to play an important role in light adaptation of photoreceptor cells. here we show that visual arrestin binds microtubules. the truncated splice variant of visual arrestin, p44, demonstrates dramatically higher affinity for microtubules than the full-length protein (p48). enhanced microtubule binding of p44 underlies it ...	2004	15272005
characterization of the human mitochondrial methionyl-trna synthetase.	human mitochondrial methionyl-trna synthetase (human mtmetrs) has been identified from the human est database. the cdna encodes a 593 amino acid protein with an 18 amino acid mitochondrial import signal sequence. sequence analysis indicates that this protein contains the consensus motifs characteristic of a class i aminoacyl-trna synthetase but lacks the zn(2+) binding motif and c-terminal dimerization region found in metrss from various organisms. the mature form of human mtmetrs has been clone ...	2004	15274629
resonance raman detection of the fe2+-c-n modes in heme-copper oxidases: a probe of the active site.	resonance raman spectroscopy has been employed to investigate the reduced cyano complexes of cytochrome aa(3) from bovine heart and rhodobacter sphaeroides and of cytochrome bo(3) from e. coli. in the aa(3)-type oxidases, the frequency of the fe-cn stretching mode is located at 468 cm(-1), and the bending fe-c-n vibration, at 500 cm(-1). the fully reduced cytochrome bo(3)-cn complex gives rise to a stretching vibration at 468 cm(-1), a bending vibration at 491 cm(-1), and a stretching c-n vibrat ...	2004	15285666
characterisation of attaching-effacing escherichia coli isolated from animals at slaughter in england and wales.	escherichia coli isolates were recovered from faecal samples taken from cattle, sheep and pigs at slaughter in england and wales. isolates (n = 1227) selected at random from this collection were each hybridised in colony dot-blot experiments with an eae gene probe that presumptively identified attaching-effacing e. coli (aeec). of the 99 (8.1%) eae positive isolates 72 were of ovine origin, 24 were of bovine origin and three of porcine origin. none were typed as o157:h7 whereas 78 were assigned ...	2004	15288926
differential effects of steroids and retinoids on bovine myelopoiesis in vitro.	pregnancy and parturition are associated with physiological changes caused by steroid hormones. alterations in number, maturity, and function of polymorphonuclear leukocytes observed in dairy cows at parturition suggest a common causative relationship with steroid hormones. this study was designed to investigate the effects of progesterone, 17-beta-estradiol, and hydrocortisone on the proliferation of bovine progenitor cells. an in vitro culturing system was used, and colonies were scored after ...	2004	15290966
sample size, library composition, and genotypic diversity among natural populations of escherichia coli from different animals influence accuracy of determining sources of fecal pollution.	a horizontal, fluorophore-enhanced, repetitive extragenic palindromic-pcr (rep-pcr) dna fingerprinting technique (hferp) was developed and evaluated as a means to differentiate human from animal sources of escherichia coli. box a1r primers and pcr were used to generate 2,466 rep-pcr and 1,531 hferp dna fingerprints from e. coli strains isolated from fecal material from known human and 12 animal sources: dogs, cats, horses, deer, geese, ducks, chickens, turkeys, cows, pigs, goats, and sheep. hfer ...	2004	15294775
rectal administration of escherichia coli o157:h7: novel model for colonization of ruminants.	escherichia coli o157:h7 causes hemorrhagic colitis and life-threatening complications. because healthy cattle are reservoirs for the bacterium, ruminant infection models have applications in analyzing the relationship between cattle and this human pathogen and in testing interventions to reduce or prevent bovine colonization with this bacterium. current approaches often do not reliably mimic natural, long-term bovine colonization with e. coli o157:h7 in older calves and adult animals (ages that ...	2004	15294790
a three-year study of enterohemorrhagic escherichia coli o157 on a farm in japan.	a long-term study was performed on the prevalence of enterohemorrhagic escherichia coli (ehec) o157 in bovine faeces. the present study was conducted on heifers raised on a farm showing a high isolation rate of ehec o157 in previous years. the prevalence of ehec o157 isolated from faecal samples was 10.6% (222/2104), 5.6% (181/3225), and 5.6% (153/2744) from 1998 to 2000, respectively. the numbers of ehec o157-positive heifers for the same 3 years were 46.3% (185/400), 36.8% (147/399), and 31.7% ...	2004	15297748
the efficacy of bovine lactoferrin in the treatment of cows with experimentally induced escherichia coli mastitis.	the effect of bovine lactoferrin (lf) was studied in experimental escherichia coli mastitis, using enrofloxacin as a comparator. mastitis was induced in six clinically healthy primiparous dairy cows by infusing 1500 colony-forming units of e. coli into a single udder quarter. the challenge was repeated into a contralateral quarter of the same cows 3 weeks later. at the first challenge, three cows were treated with 1.5 g of bovine lactoferrin intramammarily three times (12, 20 and 36 h postchalle ...	2004	15305847
effect of antibiotics in milk replacer on fecal shedding of escherichia coli o157:h7 in calves.	the objective of this study was to compare the concentration and duration of fecal shedding of escherichia coli o157:h7 between calves fed milk replacer with or without antibiotic (oxytetracycline and neomycin) supplementation. eighteen 1-wk-old holstein calves were orally inoculated with a strain of e. coli o157:h7 (3.6 x 10(8) cfu/calf) made resistant to nalidixic acid (na). rectal samples were obtained three times weekly for 8 wk following oral inoculation. fecal shedding of na-resistant e. c ...	2004	15309963
antibiotic resistance among enterotoxigenic escherichia coli from piglets and calves with diarrhea.	in vitro resistance to 8 antimicrobials among enterotoxigenic escherichia coli from piglets and calves over a 13-year period was evaluated. least resistance occurred against ceftiofur for all, followed by apramycin and gentamicin for porcine and florfenicol for bovine isolates. no significant differences were found between the first 8 and last 5 years.	2004	15317393
iron uptake by escherichia coli cultured with antibodies from cows immunized with high-affinity ferric receptors.	the synergistic effects of immunoglobulin g (igg) from cows vaccinated with ferric citrate receptor (feca) and igg from cows vaccinated with ferric enterobactin receptor (fepa) were measured in an in vitro iron uptake assay. serum was isolated and pooled within treatment from five cows each vaccinated with fepa or feca or not vaccinated. immunoglobulin g was isolated by ammonium sulfate precipitation and protein g affinity chromatography. six escherichia coli isolates from bovine intramammary in ...	2004	15328222
influence of animal origin and lineage on survival of escherichia coli o157:h7 strains in strong and weak acid challenges.	twenty-five strains of escherichia coli o157:h7 isolated from humans, cattle, and pigs were maintained in hcl (ph 2.5) and in a volatile fatty acid (vfa) mixture (ph 4.0) for up to 6 h at 37 degrees c to assess their ability to survive in acidic conditions that simulate those of the stomach and ileum, respectively. in hcl, the average group survival of bovine strains was significantly higher than that of porcine and human strains, whereas in vfas, porcine strains were significantly more resistan ...	2004	15330520
high numbers of shiga toxin-producing escherichia coli found in bovine faeces collected at slaughter in japan.	the prevalence and concentration of shiga toxin-producing escherichia coli (stec) in cattle faeces (n=605) at the time of slaughter was studied in shimane prefecture, japan on a monthly basis between april 2000 and march 2001. screening with stx-pcr determined a prevalence of 37.5%. after analysis of spread faeces and enriched samples on cefixime, tellurite and sorbitol-macconkey agar using hcl treatment, 114 stec strains were singly or concomitantly isolated from 97 cattle (15.9%). of the 605 c ...	2004	15336421
cryopreserved bovine mammary cells to model epithelial response to infection.	mammary gland epithelial cells are likely to be important effectors in defending against mastitis, yet little is known about their response mechanisms. here, we describe a cryopreserved bovine mammary epithelial cell model to study the infection response. primary cell cultures from four holstein cows were prepared, and frozen after two passages. the cell cultures from each cow were then thawed and maintained separately, yet simultaneously, and exposed to treatments that included infection with s ...	2004	15350749
changes of physicochemical indicators during mastitis and the effects of milk ejection on their sensitivity.	we examined the relationship between physicochemical indicators and somatic cells in the milk of dairy cows during experimentally induced mastitis and their significance as indicators for use in controlling udder health. we were concerned particularly with the effect of alveolar milk ejection on the sensitivity of these indicators. in expt 1, escherichia coli lipopolysaccharide (esch. coli lps) was injected into the left rear quarter to induce an inflammatory reaction in one quarter in each of s ...	2004	15354578
identification of intracellular target proteins of the calcium-signaling protein s100a12.	in this report, we have focused our attention on identifying intracellular mammalian proteins that bind s100a12 in a ca2+-dependent manner. using s100a12 affinity chromatography, we have identified cytosolic nadp+-dependent isocitrate dehydrogenase (idh), fructose-1,6-bisphosphate aldolase a (aldolase), glyceraldehyde-3-phosphate dehydrogenese (gapdh), annexin v, s100a9, and s100a12 itself as s100a12-binding proteins. immunoprecipitation experiments indicated the formation of stable complexes be ...	2004	15355353
fecal contamination of pastoral wetlands.	near-channel hill-country wetlands draining steep pastoral land in new zealand exhibit high levels of fecal contamination at a range of flows. this contamination is attributed to both the transport of bacteria into a wetland from the surrounding catchment and the direct excretion of fecal material onto wetlands by grazing cattle. e. coli concentrations observed at low to moderate flow at 20 sites varied between 0.5 x 10(1) and 2 x 10(4) most probable number (mpn) 100 ml(-1). high flow concentrat ...	2004	15356253
the incidence of shiga toxin-producing escherichia coli in cattle with mastitis in brazil.	to determine the prevalence and molecular characteristics of shiga toxin-producing escherichia coli (stec) isolates from bovine mastitic milk in brazil.	2004	15357736
the distribution of, and antibody response to, the core lipopolysaccharide region of escherichia coli isolated from the faeces of healthy humans and cattle.	there are five different core types of escherichia coli lipopolysaccharide (lps), and enterohaemorrhagic e. coli tend to have the r3 core type. it has been hypothesized that increased carriage of bacteria with a specific core type will induce higher levels of antibodies and protect against disease caused by bacteria carrying that specific lps core. approximately 320 isolates of e. coli, half from healthy human faeces and half from healthy bovine faeces have been core typed both by core-specific ...	2004	15358817
phage types and genotypes of shiga toxin-producing escherichia coli o157:h7 isolates from humans and animals in spain: identification and characterization of two predominating phage types (pt2 and pt8).	phage typing and dna macrorestriction fragment analysis by pulsed-field electrophoresis (pfge) were used for the epidemiological subtyping of a collection of shiga toxin-producing escherichia coli (stec) o157:h7 strains isolated in spain between 1980 and 1999. phage typing distinguished a total of 18 phage types among 171 strains isolated from different sources (67 humans, 82 bovines, 12 ovines, and 10 beef products). however, five phage types, phage type 2 (pt2; 42 strains), pt8 (33 strains), p ...	2004	15364983
modulating effect of serum on the stimulation of plasminogen activator inhibitor 2 production in human gingival fibroblasts by lipopolysaccharide and interleukin-1beta.	plasminogen activator inhibitor-2 (pai-2) is an important counter proteolysis factor which helps protect tissues from inflammatory stress. the expression of pai-2 can be modulated by various inflammatory stimulants and mediators. the aim of the present study was to investigate how serum factors, might modulate the effects of lipopolysaccharide (lps) and interleukin-1beta on pai-2 production by human gingival fibroblasts.	2004	15368874
comparison of a membrane surface adhesion recovery method with an ims method for use in a polymerase chain reaction method to detect escherichia coli o157:h7 in minced beef.	in this study, enrichment procedures and two recovery methods, a membrane surface adhesion technique and an immunomagnetic separation (ims), were compared for use in conjunction with a multiplex polymerase chain reaction (pcr) method with a view to describing a fast (24 h) and economical test for detection of escherichia coli o157:h7 in meat samples. the study showed no significant difference between three different enrichment media (bhi, e. coli (e.c.) broth+novobiocin, modified tryptone soya b ...	2004	15369860
a european survey of antimicrobial susceptibility among zoonotic and commensal bacteria isolated from food-producing animals.	to study antimicrobial resistance in zoonotic bacteria isolated from food animals in different countries using uniform methodology.	2004	15375107
water and ion permeation in baqp1 and glpf channels: a kinetic monte carlo study.	the kinetic monte carlo reaction-path-following technique is applied to determine the lowest-energy water pathway and the coordinating amino acids in baqp1 and glpf channels, both treated as rigid. in baqp1, water molecules pass through the pore between the asparagine-proline-alanine (npa) and selectivity filter (sf) sites one at a time. the water chain is interrupted at the sf where one water forms three stable hydrogen bonds with protein atoms. in this sf, water's conformation depends on the p ...	2004	15377535
dose dependency and individual variability of the lipopolysaccharide-induced bovine acute phase protein response.	in order to investigate the dose dependency and the individual variability of the lipopolysaccharide (lps)-induced acute phase protein response in cattle, 8 nonlactating, nonpregnant danish holstein cows were challenged 3 times each by intravenous injection of increasing doses (10, 100, and 1000 ng/kg, consecutively) of escherichia coli lps with 3-wk intervals. all 3 lps doses resulted in a rapid increase in serum concentrations of haptoglobin and serum amyloid a (saa) and a decrease in serum co ...	2004	15377612
effect of pathogen-specific clinical mastitis on milk yield in dairy cows.	our objective was to estimate the effects of the first occurrence of pathogen-specific clinical mastitis (cm) on milk yield in 3071 dairy cows in 2 new york state farms. the pathogens studied were streptococcus spp.,staphylococcus aureus, staphylococcus spp., escherichia coli, klebsiella spp., arcanobacterium pyogenes, other pathogens grouped together, and "no pathogen isolated." data were collected from october 1999 to july 2001. milk samples were collected from cows showing signs of cm and wer ...	2004	15377615
intergenic regions in the rhoptry associated protein-1 (rap-1) locus promote exogenous gene expression in babesia bovis.	members of the babesiarap-1 gene family are expressed during multiple parasite stages, and are regulated by both transcriptional and post-transcriptional mechanisms. in all babesia species, tandemly arranged rap-1 gene copies are separated by an intergenic (ig) region that is hypothesized to regulate gene expression. in this study, we tested that hypothesis by determining whether the babesia bovisrap-1 ig region could promote extra-chromosomal expression of exogenous genes introduced into merozo ...	2004	15380689
a nonradiometric, high-throughput assay for poly(adp-ribose) glycohydrolase (parg): application to inhibitor identification and evaluation.	the enzyme poly(adp-ribose) glycohydrolase (parg) catalyzes the hydrolysis of glycosidic bonds of adp-ribose polymers, producing monomeric adp-ribose units. thus, in conjunction with poly(adp-ribose) polymerase (parp), parg activity regulates the extent of in vivo poly(adp-ribosyl)ation. small molecule inhibitors of parp and parg have shown considerable promise in cellular models of ischemia-reperfusion injury and oxidative neuronal cell death. however, currently available parg inhibitors are no ...	2004	15450800
ascophyllum nodosum supplementation: a preharvest intervention for reducing escherichia coli o157:h7 and salmonella spp. in feedlot steers.	ascophyllum nodosum (tasco-14) decreased the prevalence of enterohemorrhagic escherichia coli (ehec) o157:h7 in animals fed prior to harvest. tasco-14 was supplemented on a 2% dry matter basis 14 days prior to harvest to determine its effects on ehec and salmonella spp. prevalence. two hundred mixed crossbred steers and heifers (bos indicus x bos taurus), in a large commercial finishing facility, served as experimental units. treatment (trt, n = 100) animals received a steam-rolled corn-based di ...	2004	15453570
detection and characterization of verocytotoxin-producing escherichia coli (vtec) o157 and non-o157 in cattle at slaughter.	between september 2001 to june 2002, 145 samples of bovine caecal content were collected at slaughter for verocytotoxin-producing escherichia coli (vtec) serogroups o157 and non-o157 detection. for e. coli o157 the immunomagnetic-separation technique was performed. the enterohaemolytic phenotype was the target for non-o157 vtec identification. the vero cell assay (vca) was performed for toxic activity detection. the genomic sequence for vt1, vt2 and intimin (vt1, vt2, eae genes) were identified ...	2004	15460528
distribution of escherichia coli o157 in bovine fecal pats and its impact on estimates of the prevalence of fecal shedding.	the distribution of escherichia coli o157 in bovine feces was examined by testing multiple samples from fecal pats and determining the density of e. coli o157 in immunomagnetic separation (ims)-positive fecal samples. the density of e. coli o157 in bovine feces was highly variable, differing by as much as 76,800 cfu g(-1) between samples from the same fecal pat. the density in most positive samples was <100 cfu g(-1), the limit of reliable detection by ims. testing only one 1-g sample of feces p ...	2004	15466508
optimization of an escherichia coli system for cell-free synthesis of selectively n-labelled proteins for rapid analysis by nmr spectroscopy.	cell-free protein synthesis offers rapid access to proteins that are selectively labelled with [15n]amino acids and suitable for analysis by nmr spectroscopy without chromatographic purification. a system based on an escherichia coli cell extract was optimized with regard to protein yield and minimal usage of 15n-labelled amino acid, and examined for the presence of metabolic by-products which could interfere with the nmr analysis. yields of up to 1.8 mg of human cyclophilin a per ml of reaction ...	2004	15479237
intermittent and persistent shedding of escherichia coli o157 in cohorts of naturally infected calves.	we conducted two short-term studies of cohorts of naturally infected calves to determine the prevalence and concentrations of escherichia coli o157 shed in faeces.	2004	15479421
molecular cloning of bovine cardiac muscle heat-shock protein 70 kda and its phosphorylation by camp-dependent protein kinase in vitro.	the 70-kda heat-shock protein (hsp70) has been cloned and sequenced from bovine cardiac muscle. on the basis of sequence features, the gene corresponds to the cytoplasmic form of hsp70. this cardiac hsp70 cdna clone has an open reading frame of 1926 bp coding for 641 amino acids and a predicted molecular mass of 70.25 kda. comparison of the amino acid sequence revealed an extensive sequence identity with other species of hsp70. escherichia coli expressed cardiac hsp70 stimulated a 2-fold increas ...	2004	15491140
chaperone activity and prodan binding at the self-associating domain of erythroid spectrin.	spectrin, the major constituent protein of the erythrocyte membrane skeleton, exhibits chaperone activity by preventing the irreversible aggregation of insulin at 25 degrees c and that of alcohol dehydrogenase at 50 degrees c. the dimeric spectrin and the two subunits, alpha-spectrin and beta-spectrin prevent such aggregation appreciably better, 70% in presence of dimeric spectrin at an insulin:spectrin ratio of 1:1, than that in presence of the tetramer of 25%. our results also show that spectr ...	2004	15492010
[cloning and expression of pituitary prolactin gene in ailuropoda melanoleuca].	the giant panda (ailuropoda melanoleuca) is an endangered species and indigenous to china. it has been proposed that it has a highly specialized reproductive pattern with low fecundity, but little is known about its basic reproductive biology at molecular level. in this study,the pituitary prolactin (prl) cdna of giant panda was amplified by rt-pcr from pituitary total rna and then cloned, sequenced and submitted to genbank (genbank accession no. ay161285). the sequence analysis revealed that th ...	2004	15493147
[relationship between morphological characteristics of the teat duct and prevalence of intramammary infections with streptococcus agalactiae in dairy cows].	the objective of the study was to determine the relationship between morphological findings of the surface of teat duct particularly the level of ceratosis and the prevalence of intramammary infections (imi). the study was conducted on a commercial dairy herd housing about 3000 lactating dairy cows. we examined 891 quarters in the middle of lactation. duplicate samples of quarter foremilk were collected monthly. the bacteriological status of quarters was determined according to the recommendatio ...	2004	15503535
escherichia coli o157 cluster evaluation.	we investigated a multistate cluster of escherichia coli o157:h7 isolates; pulsed-field gel electrophoresis subtyping, using a single enzyme, suggested an epidemiologic association. an investigation and additional subtyping, however, did not support the association. confirmating e. coli o157 clusters with two or more restriction endonucleases is necessary before public health resources are allocated to follow-up investigations.	2004	15504278
distribution of escherichia coli o157:h7 within and among cattle operations in pasture-based agricultural areas.	to determine the distribution of escherichia coli o157:h7 in pasture-based cattle production areas.	2004	15524323
age-related decline in carriage of ampicillin-resistant escherichia coli in young calves.	the presence of ampicillin-resistant escherichia coli (amp(r) e. coli) in the fecal flora of calves was monitored on a monthly basis in seven cohorts of calves. calves were rapidly colonized by amp(r) e. coli, with peak prevalence in cohort calves observed in the 4 months after the calves were born. the prevalence of calves yielding amp(r) e. coli in cohorts consistently declined to low levels with increasing age of the calves (p < 0.001).	2004	15528566
the sensitivity of escherichia coli strains with k1 surface antigen and rods without this antigen to the bactericidal effect of serum.	the susceptibility of escherichia coli strains with k1 surface antigen (k1+) and rods without this antigen (k1-) to the bactericidal action of normal bovine serum and human normal cord serum was determined. seventy e. coli strains (35 k1+ and 35 k1-) were isolated from urine obtained from children with urinary tract infections. the strains investigated showed variable sensitivity to the bactericidal action of the sera. e. coli k1+ strains were characterized by lower sensitivity to bactericidal e ...	2004	15530015
serum amyloid a and tnf alpha in serum and milk during experimental endotoxin mastitis.	a cross-over study was conducted to investigate the effect of intramammarily infused lipopolysaccharide (lps) on the acute phase reaction in early (el) and in late (ll) lactation. nine cows received intramammary injections of 100 microg of escherichia coli 0111:b4 lps during el and ll. the severity of each cows systemic and local signs and change in milk appearance were recorded and scored throughout the experiment. systemic and local signs were found to be more serious in el cows. tumor necrosi ...	2004	15535955
efficacy of an escherichia coli j-5 mutant strain bacterin in the protection of calves from endotoxin disease caused by subcutaneous challenge with endotoxins from escherichia coli.	the purpose of this trial was to examine the potential of a new escherichia (e) coli j-5 mutant strain bacterin to reduce the severity of clinical disease caused by subcutaneous challenge with endotoxins of gram-negative bacteria in calves. day-old to 3-day old calves (n = 40 per study phase) were randomly assigned to either of two treatment groups, i.e. a vaccinated or a placebo group. calves in the vaccinated group received an inactivated bacterin containing a j-5 mutant strain of e. coli via ...	2004	15542194
viability of milk neutrophils and severity of bovine coliform mastitis.	to study the host-pathogen interactions during escherichia coli mastitis, we first determined whether e. coli infection would change blood and milk polymorphonuclear neutrophil (pmn) chemiluminescence (cl) and viability. we then hypothesized that when e. coli invade the mammary gland, the viable pmn in milk would efficiently phagocytose and destroy e. coli before establishment of infection. we observed that the phagocytosis-dependent and independent cl were closely linked to pmn viability and we ...	2004	15545377
genetic diversity of escherichia coli recovered from the oral cavity of beef cattle and their relatedness to faecal e. coli.	to determine the genetic diversity of generic escherichia coli recovered from the oral cavities of beef cattle and their relatedness to e. coli isolated from the faeces of cattle during pasture grazing and feedlot finishing.	2004	15548305
nmr structure of the bovine prion protein isolated from healthy calf brains.	nmr structures of recombinant prion proteins from various species expressed in escherichia coli have been solved during the past years, but the fundamental question of the relevancy of these data relative to the naturally occurring forms of the prion protein has not been directly addressed. here, we present a comparison of the cellular form of the bovine prion protein isolated and purified from healthy calf brains without use of detergents, so that it contains the two carbohydrate moieties and t ...	2004	15568016
characterization of lactobacillus coryniformis dsm 20001t surface protein cpf mediating coaggregation with and aggregation among pathogens.	phenotypic characterization of aggregation phenotypes of lactobacillus coryniformis revealed that strain dsm 20001t coaggregated with escherichia coli k88, campylobacter coli, and campylobacter jejuni but not with other human pathogens. in addition, cells of these pathogens aggregated in the presence of the spent culture supernatant (scs) of strain dsm 20001t. cells of e. coli k88 remained viable in the coaggregates and aggregates for up to 24 h. both coaggregation and aggregation (co/aggregatio ...	2004	15574903
association of escherichia coli o157:h7 with houseflies on a cattle farm.	the ecology of escherichia coli o157:h7 is not well understood. the aims of this study were to determine the prevalence of and characterize e. coli o157:h7 associated with houseflies (hf). musca domestica l. hf (n = 3,440) were collected from two sites on a cattle farm over a 4-month period and processed individually for e. coli o157:h7 isolation and quantification. the prevalence of e. coli o157:h7 was 2.9 and 1.4% in hf collected from feed bunks and a cattle feed storage shed, respectively. e. ...	2004	15574966
roles of individual enzyme-substrate interactions by alpha-1,3-galactosyltransferase in catalysis and specificity.	the retaining glycosyltransferase, alpha-1,3-galactosyltransferase (alpha3gt), is mutationally inactivated in humans, leading to the presence of circulating antibodies against its product, the alpha-gal epitope. alpha3gt catalyzes galactose transfer from udp-gal to beta-linked galactosides, such as lactose, and in the absence of an acceptor substrate, to water at a lower rate. we have used site-directed mutagenesis to investigate the roles in catalysis and specificity of residues in alpha3gt tha ...	2003	14621997
effect of chemical dehairing on the prevalence of escherichia coli o157:h7 and the levels of aerobic bacteria and enterobacteriaceae on carcasses in a commercial beef processing plant.	the objective of this experiment was to test the hypothesis that cleaning cattle hides by removing hair and extraneous matter before hide removal would result in improved microbiological quality of carcasses in commercial beef processing plants. to test this hypothesis, we examined the effect of chemical dehairing of cattle hides on the prevalence of escherichia coli o157:h7 and the levels of aerobic bacteria and enterobacteriaceae on carcasses. samples from 240 control (conventionally processed ...	2003	14627275
optimization of enrichment and plating procedures for the recovery of escherichia coli o111 and o26 from minced beef.	optimization of enrichment media and selective agars for the detection of escherichia coli o26 and o111 from minced beef.	2003	14633022
antimicrobial susceptibility and factors affecting the shedding of e. coli o157:h7 and salmonella in dairy cattle.	to examine factors affecting faecal shedding of the foodborne pathogens escherichia coli o157:h7 and salmonella in dairy cattle and evaluate antimicrobial susceptibility of these isolates.	2003	14633110
implication of virulence factors in escherichia coil o157:h7 pathogenesis.	since the first documented outbreak of hemorrhagic colitis caused by escherichia coli o157:h7 in 1982, numerous publications have demonstrated or proposed putative components implicated in the pathogenesis of this gastrointestinal infection. indeed, escherichia coli o157:h7 pathogenesis is linked to several potential virulent factors such as verotoxins (or shiga-like toxins), components implicated in attaching/effacing of microvilli, and the enterohemolysin phenotypes. defining the precise molec ...	2003	14636040
the influence of organic load on the antimicrobial activity of different concentrations of naocl and chlorhexidine in vitro.	to evaluate bacterial growth after contact with sodium hypochlorite (naocl; 1 and 5%) and chlorhexidine (chx; 0.12, 0.5 and 1%) in vitro with or without the addition of organic material (bovine serum albumin (bsa) 0.5%).	2003	14641423
steroidal regulation of uterine resistance to bacterial infection in livestock.	postpartum uterine infections reduce reproductive efficiency and have significant animal welfare and economic consequences. postpartum uterine infections are classified as nonspecific, but arcanobacterium pyogenes and escherichia coli are usually associated with them in cattle and sheep. pyometra is the most common type of uterine infection in dairy cattle, and it is detected almost exclusively in cows with active corpora lutea. luteal progesterone typically down-regulates uterine immune functio ...	2003	14641941
protein refolding assisted by self-assembled nanogels as novel artificial molecular chaperone.	molecular chaperone-like activity for protein refolding was investigated using nanogels of self-assembly of cholesterol-bearing pullulan. nanogels effectively prevented protein aggregation (i.e. carbonic anhydrase and citrate synthase) during protein refolding from gdmcl denaturation. enzyme activity recovered in high yields upon dissociation of the gel structure in which the proteins were trapped, by the addition of cyclodextrins. the nanogels assisted protein refolding in a manner similar to t ...	2003	14572636
removal of endotoxin by reverse phase hplc abolishes anti-endothelial cell activity of bacterially expressed plasminogen kringle 5.	the success of recombinant protein expression/purification in escherichia coli depends on a high-fidelity system rendering purified proteins free of confounding contaminants such as endotoxin. here we report on the expression and purification of a cryptic plasminogen-derived domain, kringle 5, which was previously reported to specifically inhibit endothelial cell growth and, therefore, angiogenesis. using a histidine (his)-tag expression and ni(+)-nta agarose purification system identical to pre ...	2003	14579737
selection and characterization of peptide memitopes binding to ricin.	a combinatorial random peptide display library expressed in e. coli was employed to identify short, linear peptide sequences that showed affinity for ricin and could be used as reagents for detection and identification of ricin. one peptide, p3, from a collection of four short peptides showed specific binding to ricin. the kinetic analysis of this peptide binding to the ricin showed lower equilibrium binding constants for the peptide p3 than monoclonal antibody. this is attributed due to both sl ...	2003	14584927
increased levels of lps-binding protein in bovine blood and milk following bacterial lipopolysaccharide challenge.	several species of gram-negative bacteria, including escherichia coli, klebsiella pneumoniae, and various species of enterobacter, are common mastitis pathogens. all of these bacteria are characterized by the presence of endotoxin or lipopolysaccharide (lps) in their outer membrane. the bovine mammary gland is highly sensitive to lps, and lps has been implicated, in part, in the pathogenesis of gram-negative mastitis. recognition of lps is a key event in the innate immune response to gram-negati ...	2003	14594231
c-reactive protein and antibacterial activity in blood plasma of colostrum-fed calves and the effect of lactulose.	several milk proteins are very important for immunological defense and can be absorbed in the intestine of calves in the first hours after birth. the influence of colostrum intake and the effect of additional lactulose application on the concentration of c-reactive protein (crp) in blood were investigated. the crp is known as a mediator of innate immunity. results were compared to the bovine acute phase protein haptoglobin, and to lactalbumin, lactoferrin, and immunoglobulins in plasma from calv ...	2003	14594250
phenotypic and molecular characteristics of typical and atypical escherichia coli o157, clinical and food isolates.	enrichment, colony isolation and confirmation are three general phases of a standard diagnostic method. e. coli o 157 (the main member of ehec group) differs metabolically from other strains of e. coli in a number of ways. most isolates are slow- or non-fermenters of sorbitol and lack the enzyme beta-glucuronidase (gud). but, a variety of atypical strains of e. coli o157 (sorbitol-fermenting variants, nonmotile and gud-positive) have been reported. the discovery of these atypical pathogenic stra ...	2003	14594402
correlation between geographic distance and genetic similarity in an international collection of bovine faecal escherichia coli o157:h7 isolates.	evidence from epidemiological and molecular studies of bovine escherichia coli o157:h7 suggests that strains are frequently transmitted across wide geographic distances. to test this hypothesis, we compared the geographic and genetic distance of a set of international bovine escherichia coli o157:h7 isolates using the mantel correlation. for a measure of genetic relatedness, pulsed-field gel electrophoresis of six different restriction enzyme digests was used to generate an average dice similari ...	2003	14596534
precise and efficient cleavage of recombinant fusion proteins using mammalian aspartic proteases.	expression of recombinant proteins as translational fusions is commonly employed to enhance stability, increase solubility and facilitate purification of the desired protein. in general, such fusion proteins must be cleaved to release the mature protein in its native form. the usefulness of the procedure depends on the efficiency and precision of cleavage and its cost per unit activity. we report here the development of a general procedure for precise and highly efficient cleavage of recombinant ...	2003	14600208
immune response to a mucosally administered aflatoxin b1 vaccine.	in the present study, a mucosal vaccine was used in an effort to elicit serum igg and intestinal secretory iga against the mycotoxin aflatoxin b1 (afb) in chickens. afb was coupled to carrier proteins (bsa and porcine thyroglobulin) for use as a vaccine and elisa coating antigen, respectively. seven-day-old broiler chicks were divided into groups of 10 and immunized with one of four vaccine preparations: 1) afb-bsa conjugate alone, 2) afb-bsa linked to the b subunit of the recombinant heat-labil ...	2003	14601734
rectoanal mucosal swab culture is more sensitive than fecal culture and distinguishes escherichia coli o157:h7-colonized cattle and those transiently shedding the same organism.	enrichment and direct (nonenrichment) rectoanal mucosal swab (rams) culture techniques were developed and compared to traditional fecal culture for the detection of escherichia coli o157:h7 in experimentally infected and naturally infected cattle. holstein steers (n = 16) orally dosed with e. coli o157:h7 were sampled after bacterial colonization starting 15 days postinoculation. enrichment rams cultures (70.31% positive) were more sensitive than enrichment fecal cultures with 10 g of feces (46. ...	2003	14605119
distribution of intimin subtypes among escherichia coli isolates from ruminant and human sources.	the intimin gene eae, located within the locus of enterocyte effacement pathogenicity island, distinguishes enteropathogenic escherichia coli (epec) and some shiga toxin-producing e. coli (stec) strains from all other pathotypes of diarrheagenic e. coli. epec is a leading cause of infantile diarrhea in developing countries, and intimin-positive stec isolates are typically associated with life-threatening diseases such as hemolytic-uremic syndrome and hemorrhagic colitis. here we describe the dev ...	2003	14605134
toward an international standard for pcr-based detection of escherichia coli o157. part 1. assay development and multi-center validation.	as part of a major european research project, a diagnostic pcr assay, including an internal amplification control, was developed and validated in a collaborative trial for the detection of escherichia coli o157. the assay is based on amplification of sequences of the rfbe o157 gene. the collaborative trial, including 12 international laboratories, was carried out in two phases: phase (a) was performed with identical pcr reagents, including the internal control, provided by the sending laboratory ...	2003	14607420
dissemination of pheu- and phev-located genomic islands among enteropathogenic (epec) and enterohemorrhagic (ehec) e. coli and their possible role in the horizontal transfer of the locus of enterocyte effacement (lee).	we have recently shown that the locus of enterocyte effacement (lee) of the bovine enterohemorrhagic e. coli rw1374 (o103:h2) resides within a large pathogenicity island (pai), integrated in the vicinity of the phenylalanine trna gene phev. here we describe an additional, but lee-negative genomic island in rw1374 in the vicinity of another phenylalanine trna gene, pheu, the sequence of which is identical to phev. these two genomic islands revealed identity of the left, but a relative variability ...	2003	12635929
viability of clostridium perfringens, escherichia coli, and listeria monocytogenes surviving mild heat or aqueous ozone treatment on beef followed by heat, alkali, or salt stress.	the threat of pathogen survival following ozone treatment of meat necessitates careful evaluation of the microorganisms surviving under such circumstances. the objective of this study was to determine whether sublethal aqueous ozone treatment (3 ppm of o3 for 5 min) of microorganisms on beef surfaces would result in increased or decreased survival with respect to subsequent heat, alkali, or nacl stress. a mild heat treatment (55 degrees c for 30 min) was used for comparison. reductions in three- ...	2003	12636289
microbiological effects of hand washing at a beef carcass-breaking facility.	the hands of workers in the carcass-breaking facility at a beef packing plant were sampled by rinsing. total aerobes, coliforms, and escherichia coli were enumerated for each sample. the numbers of bacteria recovered from duplicate groups of 25 hand samples collected before and after hands were washed with an antibacterial gel, rinsed in a disinfectant solution, washed with the gel and rinsed with the disinfectant, or washed in the disinfectant for 20 s were similar for samples collected before ...	2003	12636307
rapid detection of escherichia coli o157:h7 inoculated in ground beef, chicken carcass, and lettuce samples with an immunomagnetic chemiluminescence fiber-optic biosensor.	a biosensor was evaluated with regard to its usefulness in the rapid detection of escherichia coli o157:h7 inoculated in ground beef, chicken carcass, and romaine lettuce samples. the biosensor consisted of a chemiluminescence reaction cell, a fiber-optic light guide, and a luminometer linked to a personal computer in conjunction with immunomagnetic separation. the samples inoculated with e. coli o157:h7 were first centrifuged and suspended in buffered peptone water and then incubated with anti- ...	2003	12636312
periparturient endocrine and metabolic changes in healthy cows and in cows affected by mastitis.	transition from pregnancy to lactation in dairy cows involves considerable metabolic adaptation. additional stress is incurred during infections such as periparturient mastitis. multiparous holstein-friesian cows kept under normal production conditions (n = 15) were used to evaluate changes in circulating metabolite and hormone concentrations from 5 days before to 5 days after calving. insulin-like growth factor binding protein (igfbp) profiles were also monitored. marked time-related changes we ...	2003	12650505
rapid purification of recombinant betab2-crystallin using hydrophobic interaction chromatography.	betab2-crystallin, the major subunit of beta-crystallins, is difficult to purify either from lens homogenate or from betah-or betal-crystallins. it has been prepared by heterologous expression in escherichia coli. most often, the methods used for purifying a recombinant globular protein employ the combination of ion-exchange with gel filtration chromatography. in the case of betab2-crystallin too, different approaches have been used to obtain the purified protein, majority of which use a combina ...	2003	12651120
novel bioluminescent assay of alkaline phosphatase using adenosine-3'-phosphate-5'-phosphosulfate as substrate and the luciferin-luciferase reaction and its application.	this paper describes a novel bioluminescent assay of alkaline phosphatase (alp) utilizing atp-sulfurylase and the luciferin-luciferase reaction. the principle governing the assay is as follows. adenosine-3'-phosphate-5'-phosphosulfate, which serves as the substrate for alp, is hydrolyzed enzymatically to produce adenosine-5'-phosphosulfate (aps). aps is converted into atp by atp-sulfurylase in the presence of pyrophosphate. the atp produced is detected by the luciferin-luciferase reaction. the m ...	2003	12654306
recombinant adeno-associated virus vectors efficiently and persistently transduce chondrocytes in normal and osteoarthritic human articular cartilage.	successful gene transfer into articular cartilage is a prerequisite for gene therapy of articular joint disorders. in the present study we tested the hypothesis that recombinant adeno-associated virus (raav) vectors are capable of effecting gene transfer in isolated articular chondrocytes in vitro, articular cartilage tissue in vitro, and sites of articular damage in vivo. using an raav vector carrying the escherichia coli beta-galactosidase gene (lacz) under the control of the cytomegalovirus ( ...	2003	12659680
cloning, sequencing, and heterologous expression of the murine peroxisomal flavoprotein, n1-acetylated polyamine oxidase.	the aminoacyl sequences of three regions of pure bovine n1-acetylated polyamine oxidase (pao) were obtained and used to search genbanktm. this led to the cloning and sequencing of a complete coding cdna for murine pao (mpao) and the 5'-truncated coding region of the bovine pao (bpao) gene. a search of genbanktm indicated that mpao maps to murine chromosome 7 as seven exons. the translated amino acid sequences of mpao and bpao have a -pro-arg-leu peroxisomal targeting signal at the extreme c term ...	2003	12660232