Publications

identification and neuron specific expression of the s182/presenilin i protein in human and rodent brains.	many individuals with familial alzheimer disease (fad) have mutations in a gene termed s182 or presenilin i (ps-i). currently, the ps-i gene product has not been identified and its function remains unknown. here we report that affinity purified antibodies against the predicted amino acid sequence of the ps-i gene product detected in homogenates of human, mouse, and rat brains a single antigen of approximately 48 kda. this antigen was also present in immortalized human and mouse neuronal cell cul ...	1996	8841992
internalization and translocation of a new chimeric protein composed of pseudomonas aeruginosa exotoxin a and mouse dihydrofolate reductase as a model system.	in an attempt to introduce a large peptide that is not normally translocated across membranes into the cytosol of eukaryotic cells, we created a new chimeric protein termed cedh between pseudomonas aeruginosa exotoxin a (eta) and a variant enzyme of mus musculus dihydrofolate reductase (dhfr) with reduced affinity for antifolates, eta(1-413).dhfr(1-187).eta(609-613). we have defined, genetically constructed and expressed the chimeric protein in escherichia coli. we showed that the cedh chimeric ...	1996	8844833
molecular cloning of the cdna of mouse mitochondrial nadp-dependent isocitrate dehydrogenase and the expression of the gene during lymphocyte activation.	the current report documents the molecular cloning of the mouse mitochondrial nadp-dependent isocitrate dehydrogenase (mnadp-idh) cdna. the cdna was 1,863 bp in length and contained one open reading frame encoding a 523-residue polypeptide with a predicted molecular weight of 58 kda. the cdna and the deduced amino acid (aa) sequence of the mouse mnadp-idh had a high degree of homology with those of porcine, bovine, alfalfa, and yeast. the recombinant mnadp-idh expressed in escherichia coli had a ...	1996	8867815
selective response of ternary complex factor sap1a to different mitogen-activated protein kinase subgroups.	mitogenic and stres signals results in the activation of extracellular signal-regulated kinases (erks) and stress-activated protein kinase/c-jun n-terminal kinases (sapk/jnks), respectively, which are two subgroups of the mitogen-activated protein kinases. a nuclear target of mitogen-activated protein (map) kinases is the ternary complex factor elk-1, which underlies its involvement in the regulation of c-fos gene expression by mitogenic and stress signals. a second ternary complex factor, sap1a ...	1996	8876175
involvement of two basic residues (lys-17 and arg-39) of mouse lung carbonyl reductase in nadp(h)-binding and fatty acid activation: site-directed mutagenesis and kinetic analyses.	mouse lung carbonyl reductase, a member of the short-chain dehydrogenase/reductase (sdr) family, shows a strong coenzyme preference for nadp(h) over nad(h), and is uniquely activated by fatty acids. previous chemical modification and x-ray crystallography studies show that interactions responsible for the coenzyme specificity include salt linkages between the 2'-phosphate of nadph and side-chains of lys-17 and arg-39 of the enzyme. although arg-39 is highly conserved in nadp(h)-dependent enzymes ...	1996	8889808
control by h-2 genes of the th1 response induced against a foreign antigen expressed by attenuated salmonella typhimurium.	attenuated salmonellae represent an attractive vehicle for the delivery of heterologous protective antigens to the immune system. here, we have investigated the influence of the genetic background of the host which regulates the growth and elimination of salmonella cells on the cellular response induced against a foreign antigen delivered by an aroa salmonella strain. we have tested cd4+ t-cell responses (cell proliferation and cytokine production) in various mouse strains following immunization ...	1996	8890187
the amino acids of escherichia coli enterotoxin b subunit involved in binding to bio-gel a-5m or to the glycoprotein from mouse intestinal epithelial cells.	we determined whether arg13, met31, and ser95 of the heat-labile enterotoxin b subunit (lt-b) might be involved in lt-b binding to oligosaccharides, which did not bind to the b subunit of the cholera toxin (ct-b). three lt-b mutants, r13h, m31l, and s95a were prepared by substituting three amino acid residues that differ in ct-b. these mutants formed a pentamer and exhibited the same binding ability to the gm1 ganglioside as native lt-b. although these mutants did not bind to bio-gel a-5m, they ...	1996	8890479
synthesis, processing, and oligomerization of bovine herpesvirus 1 ge and gi membrane proteins.	this study reports the identification and initial characterization of the precursors, modified forms, and oligomers of bovine herpesvirus 1 (bhv-1) gi and ge proteins with polyvalent rabbit serum specific for gi or ge. our experiments used the colorado strain of bhv-1 and mutant viruses with insertions of the escherichia coli lacz gene into the predicted ge and gi reading frames. we also translated the ge and gi open reading frames in vitro and expressed them in uninfected cells using eukaryotic ...	1996	8892910
interleukin-1beta and ibuprofen effects on cd4/cd8 cells after endotoxic challenge.	multiple-organ failure is considered a consequence of autodestructive inflammatory response during which a state of immunosuppression is produced. alterations in cd4 and cd8 lymphocytes after experimental endotoxic challenge and their correlation with the protective effects of interleukin-1beta (il-1beta) and ibuprofen pretreatment were investigated. cba/h mice were injected with lipopolysaccharide (lps) of escherichia coli (125 mg/kg); 40 mice were pretreated with il-1beta (80 ng/mouse, 24 hr p ...	1996	8895611
overcoming the crypticity of a viral t cell determinant by insertion into a chimeric bacterial protein.	among the potential t cell determinants contained in a protein antigen, the t cell response only focuses on a few immunodominant t cell determinants, whereas cryptic epitopes remain hidden to the immune system. in the present work, we have studied the antigen processing and presentation of the c3:93-115 sequences of mahoney poliovirus vp1 protein, which is immunodominant in h-2d but cryptic in h-2s and h-2q mouse mhc haplotypes. for this purpose, we genetically inserted the c3 determinant into f ...	1996	8918694
cloning and expression of the cdna for mouse nad(+)-dependent 15-hydroxyprostaglandin dehydrogenase.	the cdna for mouse nad+ dependent 15-hydroxyprostaglandin dehydrogenase (15-pgdh) was isolated from a lung cdna library. the cdna contains a 798 bp open reading frame that codes for a protein of 266 amino acids (m(r) 28775) which shares 87% identity with the human 15-pgdh protein. the regions that are believed to form the nad+ binding site and the active site are conserved in the mouse and human enzymes. the authenticity of the mouse cdna was confirmed by expression of an active 15-pgdh in esche ...	1996	8950170
adrenocortical-specific transgene expression directed by steroid hydroxylase gene promoters.	the 5'-flanking regions of genes for three mouse adrenal steroid hydroxylases were analyzed for their ability to direct adrenal cortex-specific beta-galactosidase (beta-gal) reporter expression both in cell culture and transgenic mice. the 5'-flanking regions chosen were from the genes for steroid 21-hydroxylase (21-ohase), expressed throughout the adrenal cortex and mediating both glucocorticoid and mineralocorticoid synthesis, and aldosterone synthetase (as) and steroid 11 beta-hydroxylase (11 ...	1996	8969922
removal of an intron with unique 3' branch site creates an amino-terminal protein sequence directing the scerv1 gene product to mitochondria.	the yeast scerv1 gene is of special interest because it has a dual function in mitochondrial biogenesis and in the regulation of the cell cycle. the recent discovery that the yeast scerv1 gene has a structural and functional human homologue initiated a detailed comparison of the genes and their structures. in addition the homologous alr (augmenter of liver regeneration) genes from rat and mouse have just been identified and it has been found that the mammalian proteins have a specific function i ...	1996	8972573
heterologous expression and characterisation of mouse brain fatty acid binding protein.	a novel brain-type member of the fatty acid binding protein family (b-fabp) was heterologously expressed in escherichia coli, either as inclusion bodies at 37 degrees c or in soluble form at 22 degrees c. both b-fabp renatured from inclusion bodies and the solubly expressed protein could be purified to homogeneity by anion exchange chromatography and gel filtration in a functional conformation as they bound oleic acid with high affinity. none of the five cysteines of b-fabp was involved in disul ...	1996	8722323
lipopolysaccharide stimulates differential expression of myristoylated protein kinase c substrates in murine microglia.	microglia rapidly respond to lipoplysaccharide (lps) by transformation from resting to active states and secretion of several neuro- and immuno-regulators including tumour necrosis factor alpha (tnf-alpha), interleukin 1 beta (il-1 beta), and interleukin 6 (il-6). with longer lps treatment, microglia are converted to reactive or phagocytic states with characteristics similar to macrophages in inflammation and injury processes. we have investigated lps-mediated changes in two myristoylated substr ...	1996	8723762
in vivo activities of u-100592 and u-100766, novel oxazolidinone antimicrobial agents, against experimental bacterial infections.	the upjohn oxazolidinones, u-100592 and u-100766, are orally bioavailable synthetic antimicrobial agents with spectra of activity against antibiotic-susceptible and -resistant gram-positive pathogens. in several mouse models of methicillin-resistant staphylococcus aureus infection, u-100592 and u-100766 yielded oral 50% effective doses (ed50) ranging from 1.9 to 8.0 mg/kg of body weight, which compared favorably with vancomycin subcutaneous ed50 values of 1.1 to 4.4 mg/kg. similarly, both compou ...	1996	8726028
a biological method for the quantitative measurement of tetrodotoxin (ttx): tissue culture bioassay in combination with a water-soluble tetrazolium salt.	a tissue culture bioassay, using the mouse neuroblastoma cell line (neuro2a), was improved to provide a simple and sensitive bioassay for ttx or sodium channel-blocking toxins (scb). the water-soluble tetrazolium salt, 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2h-tetraz olium, monosodium salt (wst-1), was applied to replace the time-consuming and subjective cell-counting procedure of the cells with automatic measurement, using a microplate reader. it was also confirmed that this m ...	1996	8735249
human interleukin-2 (il-2) expressed by transfected mammalian cells.	cdna for human interleukin-2 (il-2) was cloned into the prc/rsv vector for expression in animal cells. baby hamster kidney (bhk-21) cells and chinese hamster ovary (cho) cells were transfected several times using calcium phosphate and electroporation methods with the construct prc/rsv sigil2. different transfection efficiencies were obtained. the biological test on ctll-2 (mouse cytotoxic lymphocytes) showed that the kinetics of cell proliferation were different from those of ril-2 (recombinant ...	1996	8739347
searching for new tnf-alpha analogs having potential application in cancer therapy.	two new tnf-alpha analogs were prepared and tested for their anti-tumor activity on fibrosarcoma sa-1 tumor model in vivo. in analog lk-801 two histidines (his107his108) were introduced into the surface loop thus enabling efficient purification by metal-affinity chromatography. this analog showed less side effects and can serve as a lead compound to look for other useful mutations. another analog lk-802 was designed by introduction of additional pair of mutations (cys95cys148) into lk-801 in ord ...	1996	8739350
cocirculation of multiple hantaviruses in texas, with characterization of the small (s) genome of a previously undescribed virus of cotton rats (sigmodon hispidus).	an environmental and laboratory investigation was conducted after a fatal childhood case of hantavirus pulmonary syndrome occurred in deaf smith county, texas in may 1995. a trapping campaign was conducted to identify possible rodent carriers. six species of murid and heteromyid rodents were collected, and at least one hantavirus-seropositive specimen was found in each of the five murid species. tissues from a selection of 11 seropositive specimens were examined by the polymerase chain reaction ...	1996	9025697
[the dna-binding activity of the membrane protein annexin ii and its interaction with antibodies to the chromatin nuclear ribonucleoprotein (alpha-rnp)].	by screening with labeled alu dna, a clone was isolated from cdna expression library, which appeared identical in sequence to the well-known ca-phospholipid-binding protein annexin ii. to evidence the dna-binding activity of recombinant annexin ii and its presence in the cell nucleus, we have expressed full-length mouse annexin ii cdna in bacteria with pgemex vector. the expressed protein was studied with electrophoretic mobility shift assay and for its reaction with polyclonal antibody to chrom ...	1996	9045426
variation in repeat number within the alpha c protein of group b streptococci alters antigenicity and protective epitopes.	variable expression of repeating units of the protective alpha c proteins among clinical isolates of group b streptococci (gbs) may have implications for vaccine development. in this study, alpha c protein genes containing various numbers of repeats (1,2,9, and 16) were cloned in a t7 overexpression vector in escherichia coli. expression was induced by isopropyl-beta-d-thiogalactopyranoside, and proteins were purified by anion-exchange, gel filtration, or affinity chromatography or by isoelectri ...	1996	8751902
[localization of reca-like proteins in preparations of spread nuclei of mouse spermatocytes at meiotic prophase i].	earlier, polyclonal antibodies to escherichia coli reca protein were used to identify immunologically related proteins in meiotic spermatocytes of different eukaryotes. at least one such protein proved to be a component of the synaptonemal complex (sc) [1]. subsequent experiments on localization of reca-like antigens in scs of spermatocytes were performed by indirect immunocytochemical methods and electron microscopy, which showed that reca-like protein(s) at early leptotene are largely associat ...	1996	8754067
mouse and rat cystatin c: escherichia coli production, characterization and tissue distribution.	recombinant mouse (mus musculus) and rat (rattus norvegicus) cystatin c were produced by expression in escherichia coli, isolated and functionally characterized. the mouse and rat inhibitors were both fully active in titrations of papain. determination of equilibrium constants for dissociation (ki) for their complexes with the target proteinase, cathepsin b, produced values not largely different from that for human cystatin c (ki 0.07-0.13 nm). rabbit antisera against mouse and rat cystatin c we ...	1996	8761177
hypothalamic and pituitary leukemia inhibitory factor gene expression in vivo: a novel endotoxin-inducible neuro-endocrine interface.	we have recently shown expression of leukemia inhibitory factor (lif) in human fetal pituitary tissue and its in vitro induction of pomc transcription. we now use qualitative and semiquantitative rt-pcr to demonstrate that lif and lif-receptor (lif-r) are constitutively expressed in the normal mouse hypothalamus and pituitary. hypothalamic and pituitary lif and lif-r are significantly induced (up to 6- and 4-fold, respectively) in vivo in response to lipopolysaccharide endotoxin (lps) administer ...	1996	8770918
bi-directional dideoxy fingerprinting (bi-ddf): rapid and efficient screening for mutations in the big blue transgenic mouse mutation detection system.		1996	8780870
eotaxin triggers eosinophil-selective chemotaxis and calcium flux via a distinct receptor and induces pulmonary eosinophilia in the presence of interleukin 5 in mice.	understanding the processes that control selective eosinophilia is of fundamental importance in a variety of human diseases (e.g., allergies, parasitic infections, malignancy). interleukin 5, an eosinophil-specific growth and activating factor, and eotaxin appear to collaborate in this process. eotaxin is a recently described chemotactic factor that belongs to the c-c (or beta) chemokine family and has been implicated in animal and human eosinophilic inflammatory states. we have recently reporte ...	1996	8784786
in vitro binding of type 1-fimbriated escherichia coli to uroplakins ia and ib: relation to urinary tract infections.	urinary tract infections, caused mainly by escherichia coli, are among the most common infectious diseases. most isolates of the uropathogenic e.coli can express type 1 and p fimbriae containing adhesins that recognize cell receptors. while p fimbriae recognize kidney glycolipid receptors and are involved in peyelonephritis, the urothelial for type 1 fimbriae were not identified. we show that type 1-fimbriated e. coli recognize uroplakins ia and ib, two major glycoproteins of urothelial apical p ...	1996	8790381
type 1 fimbrial expression enhances escherichia coli virulence for the urinary tract.	type 1 fimbriae are adhesion organelles expressed by many gram-negative bacteria. they facilitate adherence to mucosal surfaces and inflammatory cells in vitro, but their contribution to virulence has not been defined. this study presents evidence that type 1 fimbriae increase the virulence of escherichia coli for the urinary tract by promoting bacterial persistence and enhancing the inflammatory response to infection. in a clinical study, we observed that disease severity was greater in childre ...	1996	8790416
high field epr studies of mouse ribonucleotide reductase indicate hydrogen bonding of the tyrosyl radical.	ribonucleotide reductase catalyzes by free radical chemistry the reduction of ribonucleotides to deoxyribonucleotides. the r2 protein of a class 1 ribonucleotide reductase contains a stable tyrosyl radical of neutral phenoxy character, which is necessary for normal enzymatic activity. here we present the epr spectra from the tyrosyl free radical in the r2 protein from mouse at 9.62, 115, and 245 ghz. we show that the g-value anisotropy of the mouse r2 radical, when precisely determined from high ...	1996	8798575
localization of the effector-specifying regions of gi2alpha and gqalpha.	heterotrimeric g proteins transmit hormonal and sensory signals received by cell surface receptors to effector proteins that regulate cellular processes. members of the highly conserved family of alpha subunits specifically modulate the activities of a diverse array of effector proteins. to investigate the determinants of alpha subunit-effector specificity, we localized the effector-specifying regions of alphai2, which inhibits adenylyl cyclase, and alphaq, which stimulates phosphoinositide phos ...	1996	8798740
identification of histone h2a.x as a growth factor secreted by an androgen-independent subline of mouse mammary carcinoma cells.	shionogi carcinoma 115 (sc 115) cells and chiba subline 2 (cs 2) cells are clones of an androgen-responsive mouse tumor cell line and its autonomous subline, respectively. we have shown previously that cs 2 cells produce a heparin-binding growth factor that stimulates the growth of sc 115 cells as well as the growth of themselves. in this study, a growth factor was purified from serum-free conditioned media of cs 2 cells cultured without testosterone. a heparin-binding fraction showed growth- pr ...	1996	8810267
induction by chronic autoimmune arthritis in dba/1 mice by oral administration of type ii collagen and escherichia coli lipopolysaccharide.	we have developed a new model of autoimmune arthritis in dba/1 mice by feeding chick type ii collagen (cii) for 2-3 week intervals over a 15 week period. clinically evident arthritis occurred in 8/10 mice receiving native cii (ncii; 100 micrograms/mouse) alone at 9-13 weeks. arthritis was aggravated by the further ingestion of cii, while remission occurred after withdrawal of the cii. heat-denatured cii (dcii; 200 micrograms/mouse) was also arthritogenic if co-administered with ovoinhibitor (ovi ...	1996	8810665
comparison of plasmid dna preparation methods for direct gene transfer and genetic immunization.	plasmid dna is widely used for direct gene transfer in animals to study gene therapy, gene regulation, drug delivery and genetic immunization. here we compare cesium chloride and anion-exchange purified plasmid dna for direct gene transfer in mouse muscle and show no differences in efficiency of transfection with reporter genes or in humoral response to dna-based immunization.	1996	8816242
transdominant inhibition of moloney murine leukemia virus proliferation by defective mutants of reverse transcriptase.	to investigate the dominant-negative inhibition of moloney murine leukemia virus (mo-mulv) proliferation by polymerization-defective mutants of reverse transcriptase (rt), we constructed several plasmids harboring the mo-mulv rt gene in which the yvdd sequence, one of the conserved sequences in rna-dependent polymerases, was altered, and then transformed mouse nih3t3 cells and escherichia coli with the mutant plasmids. mouse nih3t3 cells expressing these mutant rt genes were highly resistant to ...	1996	8827439
separable cis-regulatory elements that contribute to tissue- and site-specific alpha 2(xi) collagen gene expression in the embryonic mouse cartilage.	type xi collagen is a structural component of the cartilage extracellular matrix and plays an important role in skeletal morphogenesis. as a step toward defining the molecular mechanisms responsible for the regulation of type xi collagen expression, we characterized the promoter region of the mouse alpha 2(xi) collagen gene (coll1a2). we also generated transgenic mice harboring various fragments of the promoter and the first intron of coll1a2 linked to the escherichia coli beta-galactosidase gen ...	1996	8830784
survival rate of mice after transient colonization with escherichia coli clones carrying variant shiga-like toxin type ii operons.	we have previously employed a streptomycin-treated mouse model to demonstrate that escherichia coli clones expressing particular variant shiga-like toxin type ii operons differ in oral virulence, as judged by median survival time. differences in virulence were not seen between all toxin variants, including two which differed significantly in cytotoxicity for vero cells. in the present study, we have modified the animal model by withdrawing antibiotic selection and reintroducing normal mouse inte ...	1996	8831832
parameters affecting the use of the lac repressor system in eukaryotic cells and transgenic animals.	elements of the lactose operon were used to study parameters affecting gene expression in cultured cells and transgenic animals. a lac repressor protein containing a nuclear transport signal was shown to inhibit expression of a reporter gene by interacting with lac operator sequences. in cultured cells, operator sequence, operator placement and induction parameters were all shown to be important for obtaining tight repression of a reporter gene followed by high level expression upon induction. i ...	1996	8991077
dna repair functions in heterologous cells.	our genetic information is constantly challenged by exposure to endogenous and exogenous dna-damaging agents, by dna polymerase errors, and thereby inherent instability of the dna molecule itself. the integrity of our genetic information is maintained by numerous dna repair pathways, and the importance of these pathways is underscored by their remarkable structural and functional conservation across the evolutionary spectrum. because of the highly conserved nature of dna repair, the enzymes invo ...	1996	8994804
inducible retroviral vectors regulated by lac repressor in mammalian cells.	we have developed lac repressor-regulated retroviral expression vectors that are induced by beta-galactosides upon stable transduction in mammalian cells. these vectors, derived from a moloney-virus-based vector lncx, contain an internal rous sarcoma virus (rsv) or cytomegalovirus (cmv) promoter, coupled with 2 to 4 lac operator sequences and placed in anti orientation relative to the retroviral long terminal repeat (ltr). three different vectors were tested in stably infected mass populations o ...	1996	8996098
the dutpases from herpes simplex virus type 1 and mouse mammary tumour virus are less specific than the escherichia coli enzyme.	the enzyme dutpase catalyses the hydrolysis of dutp to dump and pyrophosphate, thereby suppressing incorporation of uracil into dna and providing a pool of dump, the precursor of dttp. hydrolysis of other nucleotides similar in structure to dutp would conceivably be physiologically detrimental and high specificity of the reaction seems to be necessary. in this work, we characterize the substrate specificity of the dutpases from herpes simplex virus type 1 (hsv-1) and mouse mammary tumour virus ( ...	1996	9000104
[expression of functionally active hyman cytochrome p-450c21 (cypxxia2) in escherichia coli and single-stage purification of it using metal-affinity chromatography].	active human cytochrome p-450c21 was expressed in escherichia coli and purified to homogeneity. to increase expression, cdna encoding for the n-terminal fragment of cytochrome p-450c21 was modified. four histidine codons were added to cdna encoding for the c-terminus of the protein; thus, recombinant protein could have been rapidly and effectively purified by metal-affinity chromatography. modified human cytochrome p-450c21 was expressed (40-50 nmoles/l of culture according to spectrophotometry) ...	1996	9011227
[development of genetically engineered chimeric and single chain antibodies against human brain glioma: a preliminary report].	in order to improve the clinical usefulness of mab of mouse origin in targeting diagnosis and therapy for human brain glioma, it is necessary to humanize it and reduce its molecular size. by means of rt-pcr technique, a 348 bp heavy chain variable domain (vh), and a 318 bp light chain variable domains (vl) cdna fragments were cloned from mouse hybridoma cell line sz39 secreting mab against human brain glioma. by recombinant dna technique, the two cdna fragments were linked to human igg1 heavy ch ...	1996	9387315
modulation of endotoxin-induced histamine synthesis by cytokines in mouse bone marrow-derived macrophages.	the aim of this study was to examine the roles of various cytokines in histamine synthesis by macrophages in bone marrow. pure (> 99% nonspecific esterase-, cd14-, and mac-1-positive) macrophage populations were obtained after long term culture of bone marrow cells (bone marrow-derived macrophages). culture of the cells in the presence of escherichia coli lps caused a slight, but dose-dependent, increase in histidine decarboxylase-associated histamine synthesis with a concomitant increase in the ...	1996	8543833
spatial and temporal regulation of a lacz reporter transgene in a binary transgenic mouse system.	the transgenic mouse system is a powerful tool for the study of gene function. however, when the analysis involves genes that are critical for the normal developmental process, the usefulness of transgenic mouse systems is limited (for review see hanahan, 1989; westphal and gruss, 1989; byrne et al., 1991). this is due to potential transgene interference with development in case of ectopic or high level expression. as a result, establishing permanent transgenic mouse lines expressing these types ...	1996	8589738
late administration of a lipophilic tyrosine kinase inhibitor prevents lipopolysaccharide and escherichia coli-induced lethal toxicity.	septic shock induced by gram-negative bacteria results primarily from excessive stimulation by lipopolysaccharide (lps) of macrophages to produce tumor necrosis factor (tnf)-alpha and interleukin (il)-1. the cellular effects of lps, tnf-alpha, and il-1 are mediated via tyrosine phosphorylation pathways. a recent report indicated that selective inhibitors of tyrosine kinases, tyrphostins of the ag126 family, protect mice against lps-induced lethal toxicity in mice. protection was most effective w ...	1996	8603973
mast cell modulation of neutrophil influx and bacterial clearance at sites of infection through tnf-alpha.	although mast cells have been implicated in a variety of inflammatory conditions including immediate hypersensitivity and interstitial cystitis, their physiological role in the body is unknown. we investigated the role of mast cells in host defence against bacterial infections using a well characterized mast-cell-deficiency mouse model. we report here that mast cells, which are selectively located at portals of bacterial entry, are important to host defence. mast-cell-deficient wbb6f1-w/wv mice ...	1996	8609993
activation of shiga-like toxins by mouse and human intestinal mucus correlates with virulence of enterohemorrhagic escherichia coli o91:h21 isolates in orally infected, streptomycin-treated mice.	the enterohemorrhagic escherichia coli (ehec) o91:h21 isolates b2f1 and h414-36/89 are virulent in an orally infected streptomycin-treated mouse model. previous studies demonstrated that b2f1 and h414-36/89 grow to high levels in mucus isolated from mouse small intestine and colon and that growth in small-intestine mucus is related to virulence. we measured the levels of shiga-like toxins (slts) slt-iivha and slt-iivhb produced by b2f1 after growth in luria-bertani (lb) broth supplemented with m ...	1996	8613362
cloning and characterization of the yeast hem14 gene coding for protoporphyrinogen oxidase, the molecular target of diphenyl ether-type herbicides.	protoporphyrinogen oxidase, which catalyzes the oxygen-dependent aromatization of protoporphyrinogen ix to protoporphyrin ix, is the molecular target of diphenyl ether type herbicides. the structural gene for the yeast protoporphyrinogen oxidase, hem14, was isolated by functional complementation of a hem14-1 protoporphyrinogen oxidase-deficient yeast mutant, using a novel one-step colored screening procedure to identify heme-synthesizing cells. the hem14-1 mutation was genetically linked to ura3 ...	1996	8621563
role of egr1 in regulation of stimulus-dependent cd44 transcription in b lymphocytes.	the immediate-early gene egr-1 encodes a transcription factor (egr1) that links b-cell antigen receptor (bcr) signals to downstream activation events through the regulation of previously unidentified target genes. here we identify the gene encoding the lymphocyte homing and migration protein cd44 as a target of egr1 regulation in b cells. bcr-induced increases in cd44 mrna expression and transcription levels are shown to occur in egr1-expressing but not in nonexpressing subclones of the b-cell l ...	1996	8628295
repair-deficient 3-methyladenine dna glycosylase homozygous mutant mouse cells have increased sensitivity to alkylation-induced chromosome damage and cell killing.	in escherichia coli, the repair of 3-methyladenine (3mea) dna lesions prevents alkylation-induced cell death because unrepaired 3mea blocks dna replication. whether this lesion is cytotoxic to mammalian cells has been difficult to establish in the absence of 3mea repair-deficient cell lines. we previously isolated and characterized a mouse 3mea dna glycosylase cdna (aag) that provides resistance to killing by alkylating agents in e. coli. to determine the in vivo role of aag, we cloned a large f ...	1996	8631315
vital staining of cardiac myocytes during embryonic stem cell cardiogenesis in vitro.	mouse embryonic stem (es) cells differentiate in vitro into a variety of cell types, including spontaneously contracting cardiac myocytes. the primary aim of this work was to use vital stain techniques for real-time detection of developing cardiac myocytes in es cell differentiation cultures. the -440 to +6 human cardiac alpha-actin promoter was used to direct expression of the escherichia coli reporter gene lacz (phcactlacz) into es cell-derived cardiac myocytes during cardiogenesis in vitro. u ...	1996	8635211
enhanced protection by use of a combination of anticapsule and antilipopolysaccharide monoclonal antibodies against lethal escherichia coli o18k5 infection of mice.	to study antibody-mediated protection against escherichia coli peritonitis in balb/c mice, monoclonal antibodies (mabs) were generated against the capsule (k5) and the lipopolysaccharide (o18) of e. coli. flow cytometric analysis with two selected immunoglobulin m mabs revealed that bacteria were antigenically heterogeneous. arbitrarily, three subpopulations in e. coli o18k5 cultures could be distinguished by double immunofluorescence. a subpopulation bound only the anti-k5 mab, and another subp ...	1996	8641781
induction of antigen-specific antibodies in vaginal secretions by using a nontoxic mutant of heat-labile enterotoxin as a mucosal adjuvant.	immunization of the female reproductive tract is important for protection against sexually transmitted diseases and other pathogens of the reproductive tract. however, intravaginal immunization with soluble antigens generally does not induce high levels of secretory immunoglobulin a (iga). we recently developed safe mucosal adjuvants by genetically detoxifying escherichia coli heat-labile enterotoxin, a molecule with a strong mucosal adjuvant activity, and here we describe the use of the nontoxi ...	1996	8641809
trinucleotide repeats and long homopeptides in genes and proteins associated with nervous system disease and development.	several human neurological disorders are associated with proteins containing abnormally long runs of glutamine residues. strikingly, most of these proteins contain two or more additional long runs of amino acids other than glutamine. we screened the current human, mouse, drosophila, yeast, and escherichia coli protein sequence data bases and identified all proteins containing multiple long homopeptides. this search found multiple long homopeptides in about 12% of drosophila proteins but in only ...	1996	8643671
expression in escherichia coli of sin a 1, the major allergen from mustard.	sin a 1, the major yellow mustard allergen, is a seed storage protein that belongs to the 2s albumin family. it is composed of two disulfide-bonded polypeptide chains. the cloning of this allergen has been carried out by means of the polymerase chain reaction using non-degenerate oligonucleotides encoding the n-terminal and c-terminal regions of the mature protein as primers. five genomic nucleotide sequences have been analyzed, encoding both mature polypeptide chains linked by the internal proc ...	1996	8647131
heavy chain dominance in the binding of dna by a lupus mouse monoclonal autoantibody.	antibodies h241 and 2c10 are lupus mouse igg autoantibodies that bind native dna. in previous experiments, oligonucleotide antigens affinity-labeled both h and l chains of h241 but only the h chain of antibody 2c10. primary structures of the v regions of the 2c10 h and l chains and the h241 l chain, determined from cdna, help to explain the previous affinity-labeling experiments. the 2c10 l chain cdrs had several asp residues and a net negative charge of five, whereas the 2c10 h chain cdrs had f ...	1996	8649441
presynaptic association of rad51 protein with selected sites in meiotic chromatin.	eukaryotic homologs of escherichia coli rec-a protein have been shown to form nucleoprotein filaments with single-stranded dna that recognize homologous sequences in duplex dna. several recent reports in four widely diverse species have demonstrated the association of reca homologs with meiotic prophase chromatin. the current immunocytological study on mouse spermatocytes and oocytes shows that a eukaryotic homolog, rad5l, associates with a subset of chromatin sites as early as premeiotic s phas ...	1996	8650194
the regulation of phospholipase-a2 (pla-2) by cytokines expressing hematopoietic growth-stimulating properties.	various growth factors released by macrophages and other cell types modulate normal hematopoiesis. the physiological mechanisms whereby these molecules interact with specific target cells are ill defined. eicosanoids, the products of fatty acid metabolism, are known to regulate cell proliferation and differentiation. the release of membrane-bound phospholipid by phospholipase-a2 (pla-2) is the first critical step in the initiation of membrane remodeling and eventually eicosanoid synthesis. we re ...	1996	8650256
a mouse kidney- and liver-expressed cdna having homology with a prokaryotic parathion hydrolase (phosphotriesterase)-encoding gene: abnormal expression in injured and polycystic kidneys.	to investigate abnormalities in gene expression associated with cyst formation in polycystic kidney disease, differential cdna library screening was carried out using rna from normal and cystic kidneys of the c57bl/6j-cpk mouse. among a number of genes found to be abnormally expressed was one (cdna clone 56-1) that was significantly underexpressed in cystic kidneys. hybridization analyses revealed that the 56-1 mrna is expressed primarily in kidney and liver, and that the kidney expression begin ...	1996	8654936
a bacteria-expressed mouse interleukin-1 receptor antagonist peptide protects alginate-poly-l-lysine-alginate microencapsulated rat islets against the suppressive effect of interleukin-1 beta in vitro.		1996	8658961
molecular cloning and functional identification of a plant ornithine decarboxylase cdna.	a cdna for a plant ornithine decarboxylase (odc), a key enzyme in putrescine and polyamine biosynthesis, has been isolated from root cultures of the solanaceous plant datura stramonium. reverse transcription-pcr employing degenerate oligonucleotide primers representing conserved motifs from other eukaryotic odcs was used to isolate the cdna. the longest open reading frame potentially encodes a peptide of 431 amino acids and exhibits similarity to other eukaryotic odcs, prokaryotic and eukaryotic ...	1996	8660289
btcd, a mouse protein that binds to curved dna, can substitute in escherichia coli for h-ns, a bacterial nucleoid protein.	in an escherichia coli mutant devoid of h-ns, a bacterial nucleoid protein, mouse protein btcd was able to substitute for h-ns in two tested functions. it restored cell motility and repression of the expression of the bgl operon. btcd1, a mutant btcd protein deleted of its zinc finger and thus having reduced dna binding, failed to substitute for h-ns. mouse protein btcd was shown to repress the bgl operon at the level of transcription initiation and to bind preferentially to a curved dna fragmen ...	1996	8670903
targeted disruption of the rad51 gene leads to lethality in embryonic mice.	the mouse rad51 gene is a mammalian homologue of the escherichia coli reca and yeast rad51 genes, both of which are involved in homologous recombination and dna repair. to elucidate the physiological role of rad51 protein, the gene was targeted in embryonic stem (es) cells. mice heterozygous for the rad51 null mutation were intercrossed and their offspring were genotyped. there were no homozygous (rad51-/-) pups among 148 neonates examined but a few rad51-/- embryos were identified when examined ...	1996	8692798
molecular cloning of a rat chromosome putative recombinogenic sequence homologous to the hepatitis b virus encapsidation signal.	previously, we reported that a 61-bp subgenomic hbv dna sequence (designated as 15ab, nt 1855-1915) is a hot spot for genomic recombination and that a cellular protein binding to 15ab may be the putative recombinogenic protein. in the present study, we established the existence of a 15ab-like sequence in human and rat chromosomal dna by southern blot analysis. the 15ab-like sequence isolated from the rat chromosome demonstrated a 80.9% identity with 5'-ccaagctgtgccttgggtggc-3', at 1872-1892 of t ...	1996	8692987
construction of recombinant extended single-chain antibody peptide conjugates for use in the diagnosis of hiv-1 and hiv-2.	the construction, expression and evaluation of recombinant scfv based hiv diagnostic reagents are described. in a whole-blood, erythrocyte agglutination assay format, recombinant scfv antibodies (expressed in escherichia coli), linked to a spacer domain and hiv-gp36 or -gp41 peptides, were shown to be able to detect efficiently natural antibodies against hiv in human serum. performance in trials suggests that these single chain reagents have potential as alternatives to existing fab-peptide chem ...	1996	8699008
recombinant toxoplasma gondii surface antigen 1 (p30) expressed in escherichia coli is recognized by human toxoplasma-specific immunoglobulin m (igm) and igg antibodies.	the immunodominant surface antigen of toxoplasma gondii, surface antigen 1 (sag1), was expressed in escherichia coli as a fusion protein containing a majority of the sag1 protein supplied with six histidyl residues in the n-terminal end. the recombinant protein was purified on a ni-chelate column and then on a fast-performance liquid chromatography column and was in a nonreduced condition. it was recognized by t. gondii-specific human immunoglobulin g (igg) and igm antibodies as well as by a mou ...	1996	8705683
expression of senescence-induced protein ws3-10 in vivo and in vitro.	in our efforts to characterize cellular senescence we have shown that the mrna encoding ws3-10 protein is overexpressed in senescent human diploid fibroblasts (hdf) when compared with their younger counterparts, and that forced expression of the ws3-10 cdna in young hdf results in suppression of calcium-dependent membrane currents, presumably due in part to the presence of a calcium binding domain within the ws3-10 protein. we have now expressed this protein in e. coli and have obtained affinity ...	1996	8706785
hybrid hepatitis b virus core antigen as a vaccine carrier moiety: i. presentation of foreign epitopes.	hepatitis b virus (hbv) core antigen (hbcag) is a highly immunogenic subviral particle. here, we review recent progress in the use of hbcag as a carrier moiety for heterologous epitopes. to define surface exposed and immunogenic insertion sites for foreign epitopes in hbcag, peptidic epitopes representing binding sites for virus neutralizing antibodies on the hbv surface antigens were inserted at different positions within hbcag using genetic engineering in an escherichia coli expression system ...	1996	8717391
outer membrane proteins of pseudomonas aeruginosa as vaccine candidates.	we tested the ability of the recombinant outer membrane proteins of pseudomonas aeruginosa to serve as a protective vaccine against this gram-negative pathogen in the presence of two main pathophysiological events leading to p. aeruginosa sepsis: (i) systemic infection during immunosuppression; and (ii) bacterial translocation. a hybrid vaccine was cloned which combined the protective epitopes of outer membrane protein f (oprf) and outer membrane protein i (opri). this vaccine proved to be highl ...	1996	8717398
high expression of human amelogenin in e. coli.	a human cdna, encoding for the 175-amino-acid human amelogenin, was prepared by rt pcr from tooth bud mrna and sub-cloned into pgex-kg expression plasmid for over-expression in e. coli. the expressed protein was characterized by sds-page western blotting, and n-terminal amino acid sequencing.	1996	9206336
the n-terminal region of alpha-dystroglycan is an autonomous globular domain.	the structure of the n-terminal region of mouse alpha-dystroglycan (dgn) was investigated by expression of two protein fragments (residues 30-180 and 30-438) in escherichia coli cells. trypsin susceptibility experiments show the presence of a stable alpha-dystroglycan n-terminal region (approximately from residue 30 to 315). in addition, guanidinium hydrochloride (gdn/hcl) denaturation of dgn-(30-438)-peptide, monitored by means of tryptophan fluorescence, produces a cooperative transition typic ...	1997	9210479
an upstream, dnase i hypersensitive region of the hematopoietic-expressed transcription factor gata-1 gene confers developmental specificity in transgenic mice.	the transcription factor gata-1, which is expressed in several hematopoietic lineages and multipotential progenitors, is required for the development of red blood cells and platelets. to identify control elements of the mouse gata-1 gene, we analyzed dnase i hypersensitivity of the locus in erythroid chromatin and the expression of gata-1/escherichia coli beta-galactosidase (lacz) transgenes in mice. transgenes with 2.7 kb of promoter sequences are expressed infrequently and only within adult (d ...	1997	9223298
involvement of diacylglycerol production in activation of nuclear factor kappab by a cd14-mediated lipopolysaccharide stimulus.	exposure of chinese hamster cho-k1 transfectant cells expressing mouse cd14 (cho/cd14 cells) to lipopolysaccharide (lps) induced rapid elevation of the cellular diacylglycerol (dag) and choline/phosphocholine levels and nuclear translocation of nuclear factor kappab (nfkappab). when cells were incubated with short-chain dag analogues or bacterial phospholipase c, nfkappab activation occurred even without the lps stimulus. treatment of cho/cd14 cells with tricyclo[5.2.1.0(2.6)]decyl-(9[8])xanthog ...	1997	9224650
elongation factor ts of chlamydia trachomatis: structure of the gene and properties of the protein.	a putative structural gene cluster containing four open reading frames (orfs) located downstream of the omp1 gene of chlamydia trachomatis mouse pneumonitis (mopn) was cloned and sequenced. a genbank survey indicated that the identified cluster is similar to the rpsb-tsf-pyrh(smba)-frr region of escherichia coli. the second orf was 846 bp encoding a 282-amino-acid polypeptide with a calculated m(r) 30,824. alignment of this deduced protein sequence and e. coli elongation factor ts (ef-ts, produc ...	1997	9244380
analysis of mammalian cdc37, a protein kinase targeting subunit of heat shock protein 90.		1997	9251022
heparin binding protein-44 (hbp-44)/receptor-associated protein (rap)mediates cell-substratum adhesion of mouse nih/3t3 cells through its binding to low density lipoprotein (ldl) receptor-related protein (lrp).	the low density lipoprotein receptor-related protein (lrp) is a multifunctional endocytic receptor with the ability to bind and endocytose several structurally and functionally distinct ligands. the 39 kda receptor-associated protein (rap) is an endoplasmic reticulum (er) resident protein, which is believed to function intracellularly as a molecular chaperone for lrp and to regulate its ligand binding activity along the secretory pathway. mouse heparin binding protein-44 (hbp-44) is a homologue ...	1997	9253767
the 58 kda mouse selenoprotein is a bcnu-sensitive thioredoxin reductase.	the flavoprotein thioredoxin reductase [ec 1.6.4.5] (nadph + h+ + thioredoxin-s2 --> nadp+ + thioredoxin-(sh)2) was isolated from mouse ehrlich ascites tumour (eat) cells. like the counterpart from human placenta but unlike the known thioredoxin reductases from non-vertebrate organisms, the mouse enzyme was found to contain 1 equivalent of selenium per subunit of 58 kda. the k(m) values were 4.5 microm for nadph, 480 microm for dtnb and 36 microm for escherichia coli thioredoxin, the turnover nu ...	1997	9256243
a 6-kb promoter fragment mimics in transgenic mice the prostate-specific and androgen-regulated expression of the endogenous prostate-specific antigen gene in humans.	prostate-specific antigen (psa) is a kallikrein-like serine protease, which is almost exclusively synthesized in the luminal epithelial cells of the human prostate. psa expression is androgen regulated. previously, we characterized in vitro the proximal promoter, and a strong enhancer region, approximately 4 kb upstream of the psa gene. both regions are needed for high, androgen-regulated activity of the psa promoter in lncap cells. the goal of the present study is the in vivo characterization o ...	1997	9259317
dissection of pathways implicated in integrin-mediated actin cytoskeleton assembly. involvement of protein kinase c, rho gtpase, and tyrosine phosphorylation.	a panel of antibodies to the alphaiibbeta3 integrin was used to promote adhesion of chinese hamster ovary cells transfected with the alphaiibbeta3 fibrinogen receptor. while some alphaiibbeta3 antibodies were not able to induce p125 focal adhesion kinase (p125fak) tyrosine phosphorylation, all the antibodies equally support cell adhesion but not spreading and assembly of actin stress fibers. absence of stress fibers was also obtained by plating on antibodies directed to the hamster beta1 integri ...	1997	9268301
localization of a domain in the fimh adhesin of escherichia coli type 1 fimbriae capable of receptor recognition and use of a domain-specific antibody to confer protection against experimental urinary tract infection.	the fimh subunit of type 1-fimbriated escherichia coli has been implicated as an important determinant of bacterial adherence and colonization of the urinary tract. here, we sought to localize the functionally important domain(s) within the fimh molecule and to determine if antibodies against this domain would block adherence of type 1-fimbriated e. coli to the bladder mucosa in situ and in vivo in an established mouse model of cystitis. we generated translational fusion proteins of disparate re ...	1997	9276729
novel approaches to control of bacterial infections in animals.	bacterial infections of poultry remain of great importance world-wide in terms of economic effects and public health. they include infections caused by salmonella, escherichia coli, campylobacter and pasteurella. through the introduction of rigid hygienic measures it is possible to breed and rear poultry free of these pathogens. however, the cost to the industry would be prohibitive and economically disastrous. biological measures have been introduced albeit in a relatively empirical way. antibi ...	1997	9276992
complete cdna sequence and tissue localization of n-rap, a novel nebulin-related protein of striated muscle.	we have cloned and sequenced the full-length cdna of n-rap, a novel nebulin-related protein, from mouse skeletal muscle. the n-rap message is specifically expressed in skeletal and cardiac muscle, but is not detected by northern blot in non-muscle tissues. the full-length n-rap cdna contains an open reading frame of 3,525 base pairs which is predicted to encode a protein of 133 kda. a 587 amino acid region near the c-terminus is 45% identical to the actin binding region of human nebulin, contain ...	1997	9295142
tetracycline regulated expression of vimentin in fibroblasts derived from vimentin null mice.	fibroblast cell lines were derived from vim-/- mice that express a mouse vimentin transgene in a tetracycline regulatable manner. vimentin null mouse primary embryo fibroblasts were transformed with sv-40 early genes and vim- cell lines were isolated. a vim- cell line was then serially transfected with an expression plasmid encoding the tetracycline regulatable transactivator (tta) and a mouse vimentin cdna expression plasmid under the regulation of escherichia coli tet operator and minimal cmv ...	1997	9296393
spontaneous mutation frequencies and spectra in p53 (+/+) and p53 (-/-) mice: a test of the 'guardian of the genome' hypothesis in the big blue transgenic mouse mutation detection system.	tsg-p53/big blue double transgenic mice offer a powerful tool for examining the effect of a p53 germline mutation on spontaneous somatic mutation in vivo. after sequencing the dna-binding domain of the laci gene, we previously reported no differences in mutant frequency between p53 nullizygous (-/-) and p53 wild-type (+/+) mice in liver, spleen and brain. however, jackpot mutations elsewhere in the gene may have obscured a real difference in mutation frequency and the small sample size of mutati ...	1997	9330618
in vitro and in vivo antibacterial efficacies of cfc-222, a new fluoroquinolone.	cfc-222 is a novel fluoroquinolone containing a c-7 bicyclic amine moiety with potent antibacterial activities against gram-positive, gram-negative, and anaerobic organisms. we compared the in vitro and in vivo activities of cfc-222 with those of ciprofloxacin, ofloxacin, and lomefloxacin. cfc-222 was more active than the other fluoroquinolones tested against gram-positive bacteria. cfc-222 was particularly active against streptococcus pneumoniae (mic at which 90% of isolates are inhibited [mic9 ...	1997	9333049
molecular cloning, sequencing, and expression in escherichia coli of mouse flavin-containing monooxygenase 3 (fmo3): comparison with the human isoform.	the sequence of mouse flavin-containing monooxygenase 3 (fmo3) was obtained from several clones isolated from a mouse liver cdna library. the nucleotide sequence of mouse fmo3 was 2020 bases in length containing 37 bases in the 5' flanking region, 1602 in the coding region, and 381 in the 3' flanking region. the derived protein sequence consisted of 534 amino acids including the putative flavin adenine dinucleotide and nadp+ pyrophosphate binding sites (characteristic of mammalian fmos) starting ...	1997	9344459
involvement of nitric oxide in the in vivo effects of lipopolysaccharide on the contractile and electrical properties of mouse diaphragm.	the contractile and electrical properties of the mouse diaphragm during endotoxemia were studied, and the possible role of nitric oxide (no) on these changes was investigated. the mice were injected intraperitoneally with e. coli. lipopolysaccharide (endotoxin, lps) at various times before isolation of the diaphragm to induce endotoxemia. it was observed that direct twitch tension was slightly increased, and that there was a significant increase in tetanic tension when compared with controls. th ...	1997	9349638
genes coding for enterotoxins and verotoxins in porcine escherichia coli strains belonging to different o:k:h serotypes: relationship with toxic phenotypes.	seventy-four e. coli strains isolated from piglets with diarrhea or edema disease in spain were serotyped and examined for production of heat-labile (lt) and heat-stable (st) enterotoxins (lt-i, lt-ii, stah, stap, and stb) and verotoxins (vt1, vt2, and vt2v = vte) by phenotypic (vero cell assay and infant mouse test) and genotypic (colony hybridization and pcr) methods. in general, an excellent correlation was found between the results obtained with a pcr approach and those determined with biolo ...	1997	9350767
nucleotide sequence of the porphyromonas gingivalis w83 reca homolog and construction of a reca-deficient mutant.	degenerate oligonucleotide primers were used in pcr to amplify a region of the reca homolog from porphyromonas gingivalis w83. the resulting pcr fragment was used as a probe to identify a recombinant lambda dash phage (l10) carrying the p. gingivalis reca homolog. the reca homolog was localized to a 2.1-kb bamhi fragment. the nucleotide sequence of this 2.1-kb fragment was determined, and a 1.02-kb open reading frame (341 amino acids) was detected. the predicted amino acid sequence was strikingl ...	1997	9353038
contribution of the mn-cofactored superoxide dismutase (soda) to the virulence of yersinia enterocolitica serotype o8.	enteric pathogens harbor a set of enzymes (e.g., superoxide dismutases [sod]) for detoxification of endogenous and exogenous reactive oxygen species which are encountered during infection. to analyze the role of the mn-cofactored sod (soda) in the pathogenicity of yersiniae, we cloned the soda gene of yersinia enterocolitica serotype o8 by complementation of an escherichia coli soda sodb mutant and subsequently constructed an isogenic mutant by allelic exchange. sequence analysis revealed an ope ...	1997	9353054
study of the role of the htrb gene in salmonella typhimurium virulence.	we have undertaken a study to investigate the contribution of the htrb gene to the virulence of pathogenic salmonella typhimurium. an htrb::mini-tn10 mutation from escherichia coli was transferred by transduction to the mouse-virulent strain s. typhimurium sl1344 to create an htrb mutant. the s. typhimurium htrb mutant was inoculated into mice and found to be severely limited in its ability to colonize organs of the lymphatic system and to cause systemic disease in mice. a variety of experiments ...	1997	9353064
purification and characteristics of recombinant mouse metallothionein-i from escherichia coli.	the mouse metallothionein-i (mmt-i) cdna was amplified by polymerase chain reaction (pcr), inserted into vector pgex-4t-1, and expressed in escherichia coli as a carboxyl terminal extension of the 26-kda glutathione-s-transferase (gst). analyzed by sds-page, the amount of the expressed fusion protein gst-mt was over 50% of total cellular proteins. after the fusion protein had been digested with thrombin on a glutathione-sepharose 4b affinity chromatography column, recombinant mmt-i was purified ...	1997	9354383
the design of a new mutation model for active genes: expression of the escherichia coli lac operon in mammalian cells.	the design of a novel transgenic mouse model is described that should allow analysis of mutations at a single cell level in all tissues of a model animal. the model is based on the correct regulation of the escherichia coli lac operon in mammalian cells. induction of a mutation in the laci gene will result in the loss of transcriptional repression of the lacz gene in mutated cells. expression of beta-galactosidase can subsequently be detected at the single cell level. the model was first tested ...	1997	9360635
[the curative effects of anti-tnf monoclonal antibody in e. coli infected mice].	to evaluate effects of monoclonal antibody (mcab) against tnfa on e. coli infected mice.	1997	10453575
structure of the recombinant full-length hamster prion protein prp(29-231): the n terminus is highly flexible.	the prion diseases seem to be caused by a conformational change of the prion protein (prp) from the benign cellular form prpc to the infectious scrapie form prpsc; thus, detailed information about prp structure may provide essential insights into the mechanism by which these diseases develop. in this study, the secondary structure of the recombinant syrian hamster prp of residues 29-231 [prp(29-231)] is investigated by multidimensional heteronuclear nmr. chemical shift index analysis and nuclear ...	1997	9391046
the proofreading domain of escherichia coli dna polymerase i and other dna and/or rna exonuclease domains.	prior sequence analysis studies have suggested that bacterial ribonuclease (rnase) ds comprise a complete domain that is found also in homo sapiens polymyositis-scleroderma overlap syndrome 100 kda autoantigen and werner syndrome protein. this rnase d 3'-->5' exoribonuclease domain was predicted to have a structure and mechanism of action similar to the 3'-->5' exodeoxyibonuclease (proofreading) domain of dna polymerases. here, hidden markov model (hmm) and phylogenetic studies have been used to ...	1997	9396823
identification of a novel, non-snrnp protein complex containing u1a protein.	mouse monoclonal antibodies (mabs) were generated against escherichia coli-produced u1snrnp-a (u1a) protein. u1a-specific mabs as well as mabs that reacted with both u1a and u2snrnp-b" (u2b") were isolated. mab 12e12 was unique among the characterized mabs because it failed to immunoprecipitate u1a protein produced by in vitro transcription and translation using rabbit reticulocyte lysates. however, when u1a protein was made using a wheat germ extract, mab 12e12 could immunoprecipitate u1a quite ...	1997	9404895
control of expression of plcbeta1 by lac repressor system: relationship between nuclear plcbeta1 overexpression and growth factor stimulation.	swiss 3t3 cells have a nuclear phosphoinositide signalling system which is under the control of insulin-like growth factor i (igf-i) and acts separately from that at the plasma membrane. by using the lac repressor system we were able both to obtain the inducible overexpression of phospholipase c beta1 (plc beta1) and to determine its subcellular localisation and partitioning. moreover, by comparing the level of expression at the nucleus and the percentage of cells actively incorporating bromodeo ...	1997	9405244
the 1.25 a crystal structure of sepiapterin reductase reveals its binding mode to pterins and brain neurotransmitters.	sepiapterin reductase catalyses the last steps in the biosynthesis of tetrahydrobiopterin, the essential co-factor of aromatic amino acid hydroxylases and nitric oxide synthases. we have determined the crystal structure of mouse sepiapterin reductase by multiple isomorphous replacement at a resolution of 1.25 a in its ternary complex with oxaloacetate and nadp. the homodimeric structure reveals a single-domain alpha/beta-fold with a central four-helix bundle connecting two seven-stranded paralle ...	1997	9405351