Publications

high levels of circulating neutralizing antibody in normal animals to recombinant mouse interferon-beta produced in yeast.	plasma from normal outbred swiss mice not previously given interferon (ifn) neutralized a glycosylated (high-mannose) recombinant murine ifn-beta made in yeast (rmuifn-beta y). the neutralization antibody titers were as high as 1:6,000 versus rmuifn-beta y, whereas the titers obtained with native murine ifn-beta (muifn-beta) were 100-fold less; a recombinant murine ifn-beta make in escherichia coli (rmuifn-beta ec) was not neutralized at 1:10 dilution of plasma. an elisa using rmuifn-beta y demo ...	1989	2809279
cloning and expression of a pili gene of corynebacterium renale in escherichia coli.	a plasmid gene library of corynebacterium renale piliated strain no. 109p+ was prepared in escherichia coli in order to study the chemical structure of the pili of c. renale. of 3,000 recombinant clones tested, 5 reacted with anti-pili anti-serum. the gene products of these clones reacted with anti-pili monoclonal antibodies 8/4, 5/2 and b20/3 but lacked the reactivity with 13/4. sds-page analysis revealed that the expressed protein had a molecular mass of 48 kilodalton and deletion analysis sho ...	1990	1968989
identification of plasmid-encoded mannose-resistant hemagglutinin and hep-2 and hela cell adherence factors of two diarrheagenic escherichia coli strains belonging to an enteropathogenic serogroup.	two escherichia coli strains (b/m 369 and c-35) belonging to enteropathogenic serogroup o86 were isolated from patients with infantile diarrhea and studied with respect to their cellular adherence properties. both strains exhibited adherence (ad+) to hep-2 and hela cell monolayers in vitro and expressed mannose-resistant hemagglutinating (mrha+) activity towards human, chicken, and sheep (but not mouse, rabbit, or guinea pig) erythrocytes. cellular adherence properties of both strains could be s ...	1990	1969390
evidence for positive and negative regulation of the hox-3.1 gene.	the region-specific patterns of expression of mouse homeobox genes are considered important for establishing the embryonic body plan. a 5-kilobase (kb) dna fragment from the hox-3.1 locus that is sufficient to confer region-specific expression to a beta-galactosidase reporter gene in transgenic mouse embryos has been defined. the observed reporter gene expression pattern closely parallels endogenous hox-3.1 expression in 8- to 9.5-day postcoitum (p.c.) embryos. at 10.5 days p.c. and later, the p ...	1990	1978325
nucleotide sequence of mouse hsp60 (chaperonin, groel homolog) cdna.	the cdna sequence of the 60 kda heat-shock protein from mouse 3t3 cells has been determined. the deduced amino acid sequence of mouse hsp60 protein differs from the corresponding proteins from chinese hamster and human cells in 7 and 13 residues, respectively, most of which are conservative replacements.	1990	1979012
structure, localization and function of fanf, a minor component of k99 fibrillae of enterotoxigenic escherichia coli.	the dna sequence of the k99 fanf gene, encoding fanf, was determined. an open reading frame of 999 bp was found. the primary structure of fanf was deduced and analysis revealed the presence of a signal sequence of 22 amino acid residues. the mature protein contains 311 amino acid residues (mr 33,905 d). the amino acid sequence of fanf showed similarity with the k88ab major subunit faeg. a specific mouse antiserum against fanf was prepared by constructing and purifying a hybrid cro-lacz-fanf prot ...	1990	1982454
[flow cytometric analysis of oxidative burst of phagocytes with small amount of peripheral blood].	we have developed a simple method for assessing the oxidative metabolic burst of peripheral blood leukocytes with a minute amount of whole peripheral blood by flow cytometry according to the method of bass et al. with some modification. by this method, we can measure the h2o2 production by both granulocytes and monocytes in the same blood sample. the oxidative product formation by peripheral blood neutrophils can be monitored sequentially in the same mouse infected with e. coli. the mice infecte ...	1990	2082028
isolation, characterization and expression of the complementary dna for human tumor necrosis factor (tnf-alpha).	a cdna for human tnf-alpha (615bp) was isolated by means of polymerase chain reaction (pcr) using first strand cdna from pma-induced hl-60 cells as template. the result from sequencing the 615 bp cdna fragment indicated that it corresponded to the entire sequence of mature human tnf coding region. direct expression of mature human tnf was achieved using a plasmid pht-1 constructed by ligation of the cdna and a synthetic dna. the iptg-induced bacterial product (htnf) showed cytotoxicity to mouse ...	1990	2085391
expression of the envelope antigen of dengue virus in vaccine strains of salmonella.	the envelope gene of dengue 4 virus (den) was cloned in a plasmid under the control of escherichia coli expression signals. a clone that expressed 93% of the gene was found to be detrimental to the bacterial host. another clone which carried only 76% of the e gene was found to be quite stable in vitro as well as in vivo. the killed recombinant bacteria induced antibodies in mice which recognized native den virus. attenuated salmonella typhimurium (sal) strains carrying the den-e plasmid were tes ...	1990	2101475
cell lineage analyses of epithelia and blood vessels in chimeric mouse embryos by use of an embryonic stem cell line expressing the beta-galactosidase gene.	we have established an embryonic stem (es) cell line, ms1-el4, which has the potential to make various tissues in chimeric embryos and, at the same time, expresses the beta-galactosidase gene which was introduced as a good cell marker. to examine cell behavior and lineage during embryogenesis, we injected ms1-el4 cells into host blastocysts and recovered chimeric embryos at various developmental stages. we examined the distribution of the ms1-el4 cell derivatives by staining whole embryos with x ...	1990	2112420
genomic analysis of sequence variation in tandemly repeated dna. evidence for localized homogeneous sequence domains within arrays of alpha-satellite dna.	as a model to examine the local distribution of sequence variation within large arrays of tandemly repeated dna in complex genomes, the long-range organization of alpha-satellite dna from human chromosome 17 was investigated. three individual chromosomes, representing different alpha-satellite haplotypes, were segregated into mouse and human somatic cell hybrids and their arrays sized by pulse-field gel electrophoresis. an inventory of the higher-order repeat units found in multiple separate reg ...	1990	2122000
anti-il-6 monoclonal antibodies protect against lethal escherichia coli infection and lethal tumor necrosis factor-alpha challenge in mice.	potentially fatal physiologic and metabolic derangements can occur in response to bacterial infection in animals and man. recently it has been shown that alterations in the levels of circulating cytokines such as il-6 and tnf-alpha occur shortly after bacterial challenge. to understand better the role of il-6 in inflammation, we investigated the effects of in vivo anti-mouse il-6 antibody treatment in a mouse model of septic shock. rat anti-mouse il-6 neutralizing mab was produced from splenocyt ...	1990	2124237
effects of bacterial lipopolysaccharide and calmodulin on ca2(+)-atpase and calcium in human natural killer cells, studied by a combined technique of immunoelectron microscopy and ultracytochemistry.	our previous studies indicate that bacterial lipopolysaccharide (lps) enhances natural killer (nk) cell-mediated cytotoxicity and increases intracellular calcium (ca2+) in hepatocytes. calmodulin (cam) regulates ca2(+)-atpase activity, intracellular ca2+, and is also implicated in nk cell-mediated cytolysis. in the present work, the effects of lps and cam on ca2(+)-atpase and intracellular ca2+ in human nk cells were studied by a combined technique of immunogold electron microscopy and ultracyto ...	1990	2137483
transport of the yeast atp synthase beta-subunit into mitochondria. effects of amino acid substitutions on targeting.	we have isolated the yeast atp2 gene encoding the beta-subunit of mitochondrial atp synthase and determined its nucleotide sequence. a fusion between the n-terminal 15 amino acid residues of beta-subunit and the mouse cytosolic protein dihydrofolate reductase (dhfr) was transcribed and translated in vitro and found to be transported into isolated yeast mitochondria. a fusion with the first 35 amino acid residues of beta-subunit attached to dhfr was not only transported but also proteolytically p ...	1990	2138017
the use of transgenic mice for short-term, in vivo mutagenicity testing.	in order to develop a short-term, in vivo assay to study the mutagenic effects of chemical exposure, transgenic mice were generated using a lambda shuttle vector containing a lacz target gene. following exposure to mutagens, this target can be rescued efficiently from genomic dna prepared from tissues of the treated mice using restriction minus, in vitro lambda phage packaging extract and restriction minus escherichia coli plating cultures. mutations in the target gene appear as colorless plaque ...	1990	2151115
comparison of methods in the detection of enterotoxigenic escherichia coli in a malaysian laboratory.	the prevalence of enterotoxigenic escherichia coli (etec) in 433 stool samples from diarrhoeal cases of all ages was studied using two commercially available test kits for the detection of heat labile toxin (lt) and the infant mouse assay for the heat stable toxin (st). 16 samples (3.7%) were positive for etec, of which nine were producing st alone, six lt alone and only one was producing both lt and st. although the percentage of isolation rate was low, its occurrence was almost as common as th ...	1990	2152068
cloning and sequence analyses of cdnas for interferon- and virus-induced human mx proteins reveal that they contain putative guanine nucleotide-binding sites: functional study of the corresponding gene promoter.	the human protein p78 is induced and accumulated in cells treated with type i interferon or with some viruses. it is the human homolog of the mouse mx protein involved in resistance to influenza virus. a full-length cdna clone encoding the human p78 protein was cloned and sequenced. it contained an open reading frame of 662 amino acids, corresponding to a polypeptide with a predicted molecular weight of 75,500, in good agreement with the mr of 78,000 determined on sodium dodecyl sulfate gels for ...	1990	2154602
proton-linked sugar transport systems in bacteria.	the cell membranes of various bacteria contain proton-linked transport systems for d-xylose, l-arabinose, d-galactose, d-glucose, l-rhamnose, l-fucose, lactose, and melibiose. the melibiose transporter of e. coli is linked to both na+ and h+ translocation. the substrate and inhibitor specificities of the monosaccharide transporters are described. by locating, cloning, and sequencing the genes encoding the sugar/h+ transporters in e. coli, the primary sequences of the transport proteins have been ...	1990	2172229
homologous recombination in a mammalian plasmid.	bovine papillomavirus (bpv) shuttle vectors replicate as a circular plasmid in mouse cell nuclei without impairing host cell viability. we used these vectors to analyze homologous recombination in mammalian cells. when several bpv-based plasmids carrying direct repeats were introduced into c127 cells, we detected many recombinant plasmid molecules that have lost the sequence between the repeats. many recombinant type molecules as well as parental type molecules were detected in all the cell clon ...	1990	2177135
differences in susceptibility of inbred and outbred infant mice to enterotoxigenic escherichia coli of bovine, porcine and human origin.	infant mice from outbred swiss of1 and from inbred dba/2, c57bl/6, balb/cby and cba strains were screened for usefulness in the diarrhoea model with enterotoxigenic escherichia coli (etec) strains of bovine, porcine and human origin. mouse strains were either weakly susceptible or not susceptible to etec strains of porcine or human origin bearing antigen k88, 987p, cfa/i or cfa/ii. in contrast, some mouse strains were highly susceptible to bovine and porcine etec strains bearing k99 or f41 or bo ...	1990	2179554
specific interaction of the murine transcription termination factor ttf i with class-i rna polymerases.	the 18-base-pair sequence element aggtcgaccagtactccg (the sal box) signals termination of mouse ribosomal gene transcription. this sequence is recognized by a sequence-specific dna-binding protein, ttf i, which mediates the termination of transcription by rna polymerase i (pol i). subsequently, the ends of the primary transcripts are trimmed by 10 nucleotides in a sequence-dependent 3'-terminal processing reaction. we have now investigated whether ttf i bound to its target sequence will block el ...	1990	2181320
analysis of gpt activity in mammalian cells with a chromosomally integrated shuttle vector containing altered gpt genes.	the molecular mechanisms of reversion in mammalian cells were studied utilizing the pzipgptneo shuttle vector, with the bacterial gpt gene in the vector integrated into the chromosomal dna of mouse cells. from mutant cell lines containing gpt genes with single base changes, revertants were selected for the reappearance of gpt activity. the copy number and expression of the gpt genes in such revertants were analyzed, and the gpt activity encoded by revertant genes in both mammalian cells and bact ...	1990	2181699
escherichia coli f-18 makes a streptomycin-treated mouse large intestine colonization factor when grown in nutrient broth containing glucose.	escherichia coli f-18 fima-, a type 1 fimbria-less derivative of a normal human fecal isolate, e. coli f-18, has previously been shown to be as good a colonizer of streptomycin-treated mouse large intestine as its parent, suggesting that type 1 fimbriae are not necessary in this process. in this study it was found that when e. coli f-18 fima- was grown standing overnight at 37 degrees c in nutrient broth, it remained uniformly suspended; however, when grown in nutrient broth containing 1% (wt/wt ...	1990	2182545
retropseudogenes constitute the major part of the human elongation factor 1 alpha gene family.	the elongation factor 1 alpha (ef-1 alpha) is a protein which promotes the gtp-dependent binding of aminoacyl-trna to ribosomes in the protein synthesis process. a human gene coding for ef-1 alpha has previously been cloned and sequenced along with a pseudo-gene. here, we have further analyzed the family of human ef-1 alpha genes. using an ef-1 alpha cdna as probe twelve genomic ef-1 alpha-like clones were isolated and analyzed. four of these were sequenced and found to contain ef-1 alpha retrop ...	1990	2183196
cloning and sequencing of mammalian glutathione reductase cdna.	the molecular cloning of a partial cdna to mouse glutathione reductase mrna and of a full-length cdna to the mrna of the human enzyme is described. an initial cdna clone designated lambda grm-b11 was isolated by plaque-screening of an induced mouse cdna expression library in the lambda gt11 vector with a rabbit antibody probe to human glutathione reductase. 125iodine-labelled whole anti-rabbit immunoglobulin was used as second antibody. ecori digestion of the lambda grm-b11 clone released a 720- ...	1990	2185014
evidence for two types of cytotoxic necrotizing factor in human and animal clinical isolates of escherichia coli.	we have characterized the in vitro and in vivo toxic properties of cell sonic extracts from 22 animal and human clinical isolates of escherichia coli that caused both necrosis in the rabbit skin and multinucleation in tissue cultures, two toxic properties previously reported as being specific for e. coli cytotoxic necrotizing factor (cnf). two distinct toxic phenotypes were observed. type 1, which was displayed by originally described cnf strains, was characterized by extensive multinucleation a ...	1990	2185259
escherichia coli and klebsiella vaccines and immunotherapy.	a polyvalent vaccine has been prepared from the capsular polysaccharide of 24 different serotypes of klebsiella spp. nearly 200 volunteers have received this vaccine. it is very well tolerated and elicits both binding (elisa) and functional antibody to 21 of 24 antigens. antibodies were also detected against 10 serotypes not included in the vaccine. an immunoglobulin for intravenous use (ivig) was more protective in mouse lethality assays and enhanced opsonophagocytic killing of bacteria more th ...	1990	2189000
mutagenicity evaluation of hc blue no. 1 and hc blue no. 2. iii. effects in the salmonella typhimurium/escherichia coli reversion assay and the mouse lymphoma l5178y tk+/- forward mutation assay.	the hair-dye ingredients, hc blue no. 1 (hcb1) and hc blue no. 2 (hcb2), were tested for the induction of bacterial mutation using salmonella typhimurium strains ta1535, ta1537, ta98 and ta100; and escherichia coli strains wp2uvra-. in addition, both dyes were evaluated in the mouse lymphoma l5178y tk+/- assay (mla) for the potential to induce forward mutation. a liver homogenate (s9) prepared from aroclor 1254-induced male fischer 344 rats was used to provide a means for metabolic activation. h ...	1990	2189067
expression and biochemical characterization of human immunodeficiency virus type 1 nef gene product.	nef genes from human immunodeficiency virus type 1 isolates bh10 and lav1 (lymphadenopathy-associated virus type 1) were expressed in escherichia coli under the deo operon promoter. the two proteins found in the soluble compartment of the bacterial lysate were purified by ion-exchange column chromatography to apparent homogeneity. determination of the amino-terminal sequence revealed glycine as the first amino acid in the nef protein, indicating removal of the initiator methionine during express ...	1990	2191151
[the selective isolation of cosmid clones by homologous recombination in escherichia coli--a cosmid clone containing t complex linkage dna sequence of mouse was isolated].	a procedure for the selective isolation of specific cosmid clones by homologous recombination between cosmid clones of genomic library and a probe dna sequence cloned in a plasmid in vivo has been developed. the cosmid library was constructed in a rec- host cell strain and packaged into phage particles in vivo. the rec+ host cells containing a dna sequence used as selection probe cloned in the puc plasmid were infected by packaged cosmid phage particles. there is no homology between cosmid and t ...	1990	2196913
effects of endothelin on the lethality induced by platelet-activating factor or endotoxin in the mouse.	platelet-activating factor (paf) is a mediator that decreases cardiac output and total peripheral resistances leading to profound hypotension. it seems to be involved in shock states and in the deleterious effects of endotoxin. as the natural peptide endothelin (et) shows potent vasoconstrictor properties, we evaluate its activity towards paf and endotoxin-induced lethality in mice. et, at doses which per se did not exert any effects on mice vitality, produced a dose-dependent decrease in the pa ...	1990	2208608
reverse guanine phosphoribosyltransferase selection of recombinant vaccinia viruses.	we have developed a procedure for the selection of recombinant vaccinia viruses with applicability to poxvirus mutagenesis studies and to the use of vaccinia virus as an expression vector. the method depends on the specific inability of a recombinant vaccinia virus expressing the escherichia coli guanine phosphoriboxyltransferase gene (gpt) to form plaques on a hypoxanthine-guanine phosphoribosyltransferase-negative line of mouse fibroblasts in the presence of 6-thioguanine. recombinant viruses ...	1990	2219714
embryonic neural cell adhesion molecule in cerebrospinal fluid of younger children: age-dependent decrease during the first year.	poly-alpha-2,8-n-acetylneuraminic acid (poly-alpha-2,8-neuac) is developmentally expressed in neural tissue of higher animals, where it is covalently attached to the neural cell adhesion molecule (ncam), a large integral membrane glycoprotein mediating cell-cell adhesion during neuronal development. ncam exists in several molecular forms, of which only embryonic ncam carries lengthy chains (n greater than 5) of poly-alpha-2,8-neuac. chemically identical poly-alpha-2,8-neuac of bacterial origin i ...	1990	2230809
molecular cloning, sequencing, and expression of mouse ferrochelatase.	the cdna encoding mouse ferrochelatase (protoheme ferrolyase, ec 4.99.1.1) was isolated from a mouse erythroleukemia (mel) cell cdna library in lambda gt11 expression vector, by immunoscreening with a polyclonal antibody. two full-length clones containing cdna inserts of 2.2 and 2.90 kilobases were obtained. these clones have the same entire enzyme coding region, but alternative putative polyadenylation sites in the 3'-noncoding regions. from the deduced primary structure, a putative leader sequ ...	1990	2246229
interactions of escherichia coli and proteus mirabilis with mouse mononuclear phagocytes.	five strains of enterobacteria (three of escherichia coli and two of proteus mirabilis) were studied to assess and compare their phagocytic uptake and intracellular killing by mouse macrophages. each strain was injected intraperitoneally into separate groups of mice and peritoneal exudate cells were harvested after 3 min for phagocytosis to occur in vivo. acridine orange staining showed that there were approximately 10-fold fewer intracellular p. mirabilis than e. coli cells. the average numbers ...	1990	2250285
hepatotoxic activity of campylobacter jejuni.	hepatotoxic factor(s) were isolated from whole-cell lysates of campylobacter jejuni gifu 8734 and purified by chromatography. a single intravenous injection of 10 micrograms of this factor reproducibly produced hepatitis in mice, as determined by histology and liver function tests. the hepatic lesions were very similar to those evoked by c. jejuni infection. tissue-culture studies with mouse hepatocytes demonstrated that low concentrations of the factor caused release of hepatic enzymes into the ...	1990	2250287
heat shock response of murine chlamydia trachomatis.	we have investigated the heat shock response in the mouse pneumonitis strain of chlamydia trachomatis. the kinetics of the chlamydial heat shock response resembled that of other procaryotes: the induction was rapid, occurring over a 5- to 10-min time period, and was regulated at the level of transcription. immunoblot analysis and immunoprecipitations with heterologous antisera to the heat shock proteins dnak and groel demonstrated that the rate of synthesis, but not the absolute amount of these ...	1990	2254267
detection of the aerolysin gene in aeromonas hydrophila by the polymerase chain reaction.	synthetic oligonucleotide primers were used in a polymerase chain reaction (pcr) technique to detect the gene for aerolysin in strains of aeromonas hydrophila and to screen for identical genes in a. caviae, a. sobria, and a. veronii isolated from patients with diarrheal disease. primers targeted a 209-bp fragment of the aer gene coding for the beta-hemolysin and detected template dna only in the pcr using nucleic acid (na) from hemolytic strains of a. hydrophila which were also cytotoxic to vero ...	1990	2254423
association of virulence markers with animal pathogenicity of escherichia coli in different models.	employing chicken and several strains of mice, different routes (intraperitoneal, subcutaneous) of infections and isogenic pairs of strains, association of virulence markers with animal pathogenicity was studied in escherichia coli. mouse virulence of avian strains was less significant than the lethality for chicks of human strains. ld50 in various animals did not differ significantly. strains with antigen k1 were more virulent for mice than their k1- derivatives. loss of haemolysin (hly), manno ...	1990	2270740
thrombopoietic activity of human interleukin-6.	thrombopoietin (tpo), a regulatory factor in platelet production, was purified from the conditioned medium of tnk-01 cells cultured in the presence of human interleukin-1. the n-terminal sequence of purified tpo was determined to be vppgedskdvaaphrqplt, identical to that of the n-terminal region of human interleukin-6 (il-6). two forms of tpo with molecular masses of 24 and 27 kda were identified as il-6 by western analysis using an anti-il-6 antibody. commercial recombinant human il-6 produced ...	1990	2298297
expression and secretion of aequorin as a chimeric antibody by means of a mammalian expression vector.	a fusion protein has been expressed from the relevant genes in mammalian cells consisting of the photoprotein aequorin and an anti-4-hydroxy-3-nitrophenacetyl antibody gene. this chimeric antibody has allowed the development of a sensitive luminescent immunoassay. initially the cdna of the photoprotein aequorin from aequorea victoria was cloned and expressed in escherichia coli. the gene was expressed as apoaequorin and, by using luciferin isolated from renilla reniformis, its activity was found ...	1990	2315301
glucose phosphorylation in tumor cells. cloning, sequencing, and overexpression in active form of a full-length cdna encoding a mitochondrial bindable form of hexokinase.	in rapidly growing tumor cells exhibiting high glucose catabolic rates, the enzyme hexokinase is markedly elevated and bound in large amounts (50-80% of the total cell activity) to the outer mitochondrial membrane (arora, k.k., and pedersen, p.l. (1988) j. biol. chem. 263, 17422-17428; parry, d.m., and pedersen, p.l. (1983) j. biol. chem. 258, 10904-10912). in extending these studies, we have isolated a cdna clone of hexokinase from a lambda gt11 library of the highly glycolytic, c37 mouse hepat ...	1990	2318862
the primary structure of rat ribosomal protein s16.	the amino acid sequence of rat ribosomal protein s16 was deduced from the sequence of nucleotides in a recombinant cdna and confirmed from the nh2-terminal amino acid sequence of the protein. s16 contains 145 amino acids (the nh2-terminal methionine is removed after translation of the mrna) and has a molecular mass of 16,304. hybridization of the cdna to digests of nuclear dna suggests that there are 11-13 copies of the s16 gene. the mrna for the protein is about 700 nucleotides in length. rat s ...	1990	2332055
the gene sequence and some properties of protein h. a novel igg-binding protein.	the gene for protein h, a novel bacterial cell wall protein with specific affinity for human igg fc, was cloned from a group a streptococcus and expressed in escherichia coli. recombinant e. coli cells produced two forms of a human igg fc-binding protein, one with an apparent mr of 42 kda in a periplasmic fraction and the other with an apparent mr of 45 kda in a mixed fraction of cytoplasms and membranes. both 42-kda and 45-kda protein preparations similarly bound to human igg1 to igg4, human ig ...	1990	2332638
interleukin 6 perfusion stimulates reconstitution of the immune and hematopoietic systems after 5-fluorouracil treatment.	effects of interleukin 6 (il-6) on the functional capacity of the immune and hematopoietic systems in 5-fluorouracil (5-fu)-treated mice were determined. il-6 (5 x 10(4) units/mouse/day) was administered s.c. for 7 days by implantation of an osmotic pump, since it was demonstrated that a much higher increase in the primary response to sheep rbc was observed by administration of slowly released rather than daily s.c. injection of il-6. il-6 perfusion significantly augmented anti-sheep rbc antibod ...	1990	2334892
detection of dna strand breaks in hela cells in vitro and in mouse sarcoma 180 cells in vivo induced by an alkylating agent, carboquone, using in situ nick translation.	we determined the correlation between dna strand breaks and the toxicity of carboquone (cq) in hela cells in vitro and in mouse sarcoma 180 (s-180) cells in vivo, using in situ nick translation. the break sites in the dna were translated artificially in the presence of escherichia coli dna polymerase i and [3-h]-labelled dttp, and sites in the dna were visualized by autoradiographic observation of grains in the nuclei. these breaks appeared as early as 5 min in the cq-treated hela cells and incr ...	1990	2342771
27 amino acid residues can be deleted from the n-terminus of human lymphotoxin without impairment of its cytotoxic activity.	in order to study the relationship between activity and structure of human lymphotoxin (hlt, 171 aa), we synthesized the gene (519 bp) for hlt and expressed it in escherichia coli. purification of the recombinant hlt from crude extracts was difficult because of the low level of expression of the gene. to improve the yield of the recombinant protein, we prepared five truncated genes for mutant proteins in which 25, 26, 27, 28 and 37 amino acid residues, respectively, were missing from the n-termi ...	1990	2361063
isolation and structural characterization of a cdna clone encoding the human dna repair protein for o6-alkylguanine.	o6-methylguanine-dna methyltransferase (mgmt; dna-o6-methylguanine:protein-l-cysteine s-methyltransferase, ec 2.1.1.63), a unique dna repair protein present in most organisms, removes the carcinogenic and mutagenic adduct o6-alkylguanine from dna by stoichiometrically accepting the alkyl group on a cysteine residue in a suicide reaction. the mammalian protein is highly regulated in both somatic and germ-line cells. in addition, the toxicity of certain alkylating drugs in tumor and normal cells i ...	1990	2405387
comparison of biological and immunological activities of human monocyte-derived interleukin 1 beta and human recombinant interleukin 1 beta.	recombinant human interleukin 1 beta (rhil-1 beta) and supernatants of escherichia coli lipopolysaccharides-stimulated human monocyte (mo) cultures, containing native human il-1 beta (nhil-1 beta), demonstrate significant differences when tested in the mouse co-stimulatory thymocyte (lymphocyte activating factor [laf]) assay. the aims of the present study were to investigate this characteristic difference between rhil-1 beta and mo culture supernatants (mo supernatants), and to compare the biolo ...	1990	2408138
direct gene transfer into mouse muscle in vivo.	rna and dna expression vectors containing genes for chloramphenicol acetyltransferase, luciferase, and beta-galactosidase were separately injected into mouse skeletal muscle in vivo. protein expression was readily detected in all cases, and no special delivery system was required for these effects. the extent of expression from both the rna and dna constructs was comparable to that obtained from fibroblasts transfected in vitro under optimal conditions. in situ cytochemical staining for beta-gal ...	1990	1690918
the fine specificity of anti-la antibodies induced in mice by immunization with recombinant human la autoantigen.	because of increasing evidence suggesting that anti-la autoantibodies are induced in humans by an ag-specific mechanism, we investigated the antibody response of animals immunized with the human la ag and studied its relationship to the anti-la response of autoimmune patients. anti-la antibodies were raised in 6- to 8-wk-old male mrl(-)+/+, c57bl/6j, balb/c, and a/j mice by immunizing with authentic human la protein obtained by recombinant expression in escherichia coli. as we have shown previou ...	1990	1692063
identification and mapping of an immunogenic region of mycoplasma hyopneumoniae p65 surface lipoprotein expressed in escherichia coli from a cloned genomic fragment.	a previously characterized lipid-modified amphiphilic surface protein of mycoplasma hyopneumoniae, p65, has been defined by its reaction with a surface-binding monoclonal antibody (mab) and by its exclusive partitioning into the detergent phase during triton x-114 phase fractionation (k. s. wise and m. f. kim, j. bacteriol. 169:5546-5555, 1987). in the current study, polyclonal mouse antibody (pab) to gel-purified p65 was used to identify recombinant phage plaques expressing p65-related epitopes ...	1990	1695206
synthesis and expression in escherichia coli of dna encoding the murine lambda 1 chain of a monoclonal antibody specific for salmonella serotype b o-antigen.	a 658 bp dna sequence corresponding to the murine lambda 1 chain of a monoclonal antibody, se155-4, specific for the salmonella serotype b o-antigen, was designed using escherichia coli preferred codons and chemically synthesized by ligation of synthetic fragments into a linearized plasmid followed by transformation into e. coli. a synthetic signal peptide (ompa) was fused to express the l chain as a free polypeptide into the periplasm of e. coli cells. after isolation and purification, heterolo ...	1990	1695376
patterns of expression of position-dependent integrated transgenes in mouse embryo.	the abilities to introduce foreign dna into the genome of mice and to visualize gene expression at the single-cell level underlie a method for defining individual elements of a genetic program. we describe the use of an escherichia coli lacz reporter gene fused to the promoter of the gene for hypoxanthine phosphoribosyl transferase that is expressed in all tissues. most transgenic mice (six of seven) obtained with this construct express the lacz gene from the hypoxanthine phosphoribosyltransfera ...	1990	1696727
antigenic conservation of the 15,000-dalton outer membrane lipoprotein pcp of haemophilus influenzae and biologic activity of anti-pcp antisera.	a gene from haemophilus influenzae encoding an outer membrane lipoprotein of about 15,000 daltons and which comigrates with the peptidoglycan-associated lipoprotein (pal) of h. influenzae on sodium dodecyl sulfate-polyacrylamide gel electrophoresis has been previously reported and designated pcp gene, and its product has been designated pcp. in order to obtain specific immunologic probes for the analysis of pcp expression, cellular location, and antigenic conservation in h. influenzae, pcp was f ...	1990	1698180
aromatic-dependent salmonella as live vaccine presenters of foreign epitopes as inserts in flagellin.	synthetic oligonucleotides specifying amino acid sequences identified as epitopes of various foreign antigens (cholera toxin subunit b, hepatitis b surface protein and others) have been inserted at an ecorv-ecorv deletion site in a cloned salmonella flagellin gene; the resulting plasmids, when placed in flagellin-negative escherichia coli or salmonella sp. strains, caused production of flagellin expressing the epitope. if the chimeric flagellin allowed formation of flagella, the epitope was expo ...	1990	1714093
production of a protein a--lymphotoxin hybrid protein for cancer-targeted therapy.	a hybrid protein between staphylococcal protein a and human lymphotoxin, alt, was produced in escherichia coli by expression of a recombinant plasmid containing the respective genes. igg-binding activity of alt was confirmed by western blotting analysis and by affinity purification with igg-sepharose column chromatography. the purified alt had cytotoxicity on mouse l-929 cells and its specific activity was approximately 3.5-5.0 x 10(6) u mg-1. alt was partially degraded by a protease including i ...	1990	1366561
immunological characterization of in vitro forms of homoserine dehydrogenase from carrot suspension cultures.	multiple forms of homoserine dehydrogenase (hsdh) from carrot (daucus carota l.) have been identified. one form of hsdh (t-form) has a relative molecular weight of 240,000 and is strongly inhibited by threonine. another form (k-form) has a relative molecular weight of 180,000 and is insensitive to inhibition by threonine. the interconversion of these two forms is dependent upon the presence or absence of threonine and potassium. polyacrylamide electrophoretic gels stained for hsdh activity and p ...	1990	16667288
purification and characterization of the cytokine-induced macrophage nitric oxide synthase: an fad- and fmn-containing flavoprotein.	a soluble nitric oxide (no) synthase activity was purified 426-fold from a mouse macrophage cell line activated with interferon gamma and bacterial lipopolysaccharide by sequential anion-exchange, affinity, and gel filtration chromatography. sds/page of the purified no synthase gave three closely spaced silver-staining protein bands between 125 and 135 kda. when assayed in the presence of l-arginine, nadph, tetrahydrobiopterin, fad, and reduced thiol, purified no synthase had a specific activity ...	1991	1715579
immunoadjuvant activity of oral lactobacillus casei: influence of dose on the secretory immune response and protective capacity in intestinal infections.	lactobacilli, often used as effectors of host functions, could play an important role in maintaining human health by controlling other intestinal microorganisms capable of producing harmful effects. using an experimental model, we studied the effect of different oral doses of lactobacillus casei on the secretory iga response and the protective capacity of the microorganism in preventing intestinal infections. the optimization of the protective dose of lb. casei by previous feeding and the use of ...	1991	1722492
constitutive production of granulocyte colony-stimulating factor by hybrids of a sv40-transformed mouse macrophage and a renal adenocarcinoma cell line.	mouse macrophage bam3 cells produced colony-stimulating factors (csfs) after stimulation with bacterial lipopolysaccharide (lps). by assaying the csf using various interleukin 3-dependent cell lines, it was shown that most of the csfs produced by bam3 cells were granulocyte csf (g-csf). the granulocyte-macrophage csf (gm-csf) gene was also expressed in bam3 cells after stimulation with lps. when bam3 cells were fused with the mouse renal adenocarcinoma cell line rag which does not produce g-csf, ...	1991	1723285
cloning, expression and characterization of the human transcription elongation factor, tfiis.	the cdna for the human elongation factor, tfiis, has been cloned and expressed in e. coli with the t7 expression system. this 280-amino acid tfiis protein is shorter by 21 residues than that of the mouse. the missing 21 residues are located in the amino-terminal region, which is not thought to be required for transcriptional stimulation. apart from this gap, human and mouse proteins reveal 96% overall identity and 98.5% sequence similarity if conservative substitutions are taken into account. th ...	1991	1708494
production of a recombinant human t-cell leukemia virus type-i trans-activator (tax1) antigen and its utilization for generation of monoclonal antibodies against various epitopes on the tax1 antigen.	a 42-kda recombinant protein, px141, consisting of the trans-activator protein encoded by human t-cell leukemia virus (htlv-1) (tax1 antigen) and the amino-terminal fusion peptide of 12 amino acid residues of the alpha-peptide encoded by the plasmid puc19 was produced. in order to investigate the immunogenicity of the tax1 antigen, mice were immunized with the purified px141 and 4 anti-tax1 monoclonal antibodies (mabs) designated taxy-1, taxy-6, taxy-7 and taxy-8 were generated, and their reacti ...	1991	1710610
synthesis and expression in escherichia coli of cistronic dna encoding an antibody fragment specific for a salmonella serotype b o-antigen.	a 1460-bp dna encoding the two chains of the antigen-binding fragment (fab) portion of a monoclonal antibody have been chemically synthesized and expressed in escherichia coli. the antibody, se155-4, is specific for a salmonella serogroup b o-antigen and its crystal structure is under investigation. the genes were synthesized according to a strategy that allows for easy manipulation in genetic engineering studies of the fab-binding site. each gene is preceded by the ompa secretory signal and a r ...	1991	1711496
expression, purification, and crystallization of the adipocyte lipid binding protein.	the murine adipocyte lipid binding protein (albp/ap2) has been cloned and expressed in escherichia coli, purified to homogeneity, biochemically characterized, and crystallized for x-ray diffraction study. in the cloning, the albp coding region was placed under control of the reca promoter and downstream of the phage t7 g-10 translation enhancer sequence. nalidixic acid (50 micrograms/ml) induced the expression of albp 20-fold over that attained using the pt7 system previously reported (chinander ...	1991	1650358
altering the conserved nucleotide binding motif in the salmonella typhimurium muts mismatch repair protein affects both its atpase and mismatch binding activities.	the salmonella typhimurium and escherichia coli muts protein is one of several methyl-directed mismatch repair proteins that act together to correct replication errors. muts is homologous to the streptococcus pneumoniae hexa mismatch repair protein and to the duc1 and rep1 proteins of human and mouse. homology between the deduced amino acid sequence of both muts and hexa, and the type a nucleotide binding site consensus sequence, suggested that atp binding and hydrolysis play a role in their mis ...	1991	1651234
nucleoside diphosphate kinase from escherichia coli; its overproduction and sequence comparison with eukaryotic enzymes.	the gene encoding nucleoside diphosphate (ndp) kinase of escherichia coli was identified by polymerase chain reaction using oligodeoxyribonucleotide primers synthesized on the basis of consensus sequences from myxococcus xanthus and various eukaryotic ndp kinases. the gene (ndk), mapped at 54.2 min on the e. coli chromosome, was cloned and sequenced. the e. coli ndp kinase was found to consist of 143 amino acid residues that are 57, 45, 45, 42, 43, and 43% identical to the m. xanthus, dictyostel ...	1991	1657712
interferon-inducible mouse mx1 protein that confers resistance to influenza virus is gtpase.	the murine mx1 protein is an interferon-inducible nuclear protein and confers resistance to influenza virus infection even though the resistance mechanism is yet unclear. the mx1 protein contains a tripartite gtp-binding domain consisting of gxxxxgks, dxxg, and t/nkxd motifs. in the gtpase gene superfamily such as p21ras protein, signal-transducing g protein, and translation elongation factor, the gtpase activity plays a key role in each protein function. here we show that gtpase activity is ind ...	1991	1657964
unusual cell specific expression of a major human cytomegalovirus immediate early gene promoter-lacz hybrid gene in transgenic mouse embryos.	transgenic mice carrying the human cytomegalovirus immediate early gene promoter driving the e. coli lacz gene displayed an unusual cell specific expression of beta-galactosidase during development. lacz expression was first detected in cells lining the apex of the neural fold of day 8.5 embryos. by day 10 of gestation, expression was prominent in the spinal ganglia, the ganglia of cranial nerves v, vii, viii, ix, and x, in a line of cells marking the ventrolateral pathway adjacent to the dermam ...	1991	1659441
expression of escherichia coli f-18 type 1 fimbriae in the streptomycin-treated mouse large intestine.	escherichia coli f-18, isolated from the feces of a healthy human, makes type 1 fimbriae and is an excellent colonizer of the streptomycin-treated mouse large intestine. recently, it was shown that the inability to produce type 1 fimbriae had no effect on the ability of e. coli f-18 to colonize the streptomycin-treated mouse large intestine, suggesting the possibility that e. coli f-18 does not express type 1 fimbriae in vivo. however, we show here that e. coli f-18 does express type 1 fimbriae ...	1991	1672304
expression of mouse tyrosine hydroxylase in escherichia coli.	enzymatically active mouse tyrosine hydroxylase (th) was successfully expressed at a high level in escherichia coli using a t7 rna polymerase directed expression system. the specific activity of mouse th in e. coli cell lysate was 7.5 nmol/mg protein/min. kinetic characteristics of recombinant th were examined. km for tyrosine and (6r)-tetrahydrobiopterin (6r-bh4) cofactor were determined to be 7.2 microm (420 microm 6r-bh4), 19 microm [( 6r-bh4] less than 55 microm, 20 microm tyrosine) and 54 m ...	1991	1677565
protective effect of tissue ferritins in experimental escherichia coli infection of mice in vivo.	the effect of ferritins from horse (fh) and bovine (fb) spleen and murine liver (fm) on the survival rate of cfw mice lethally infected with escherichia coli (strain 8440-78 k 80/b) was evaluated. ferritins given intravenously 24 h before intravenous inoculation of bacteria, protected mice most effectively from death due to infection. the effect was dose dependent. at 500 micrograms of ferritin per mouse, the maximum survival rates were 86% (fh), 81% (fm) and 79% (fb), while only 5% of the contr ...	1991	1768608
[cloning of the gene and immunochemical specificity of recombinant immunoglobulin binding protein v7 from streptococcus valente (group g)].	the fragment of the structural gene coding for the fc-receptor of streptococcus valente (g group) has been cloned. the resulting recombinant plasmid ppgsv1 contains the o, kb hindiii fragment of streptococcal chromosomal dna inserted into the vector plasmid puc19 and determines the expression of the 31 kd protein in escherichia coli cells. the protein binds the immunoglobulins of human, rabbit, guinea pig origin, but in contrast to the g protein of another g group streptococcus it is nonreactive ...	1991	1784298
[cloning and detailed mapping of the fra-ymt region of the yersinia pestis pfra plasmid].	the genetical libraries of the pfra plasmid of yersinia pestis genes were obtained by insertion into the psti, salgi, ecori, xhoi restriction sites of the cosmid vector phc79. the immunochemical analysis of the recombinant clones has revealed the clones synthesizing the antigen fl (fraction i) and mouse toxin (ymt--yersinia pestis murine toxin). the restriction analysis of the plasmids from antigen synthesizing clones has permitted to construct the detailed physical map of the fra-ymt region of ...	1991	1787840
positive regulator for the expression of mba protein of the virulence plasmid, pkdsc50, of salmonella choleraesuis.	a positive regulator was identified within a 2.3 kb fragment of the 6.4 kb mouse bacteremia region (mba region) of the virulence pkdsc50 plasmid of salmonella choleraesuis. sodium dodecyl sulphate polyacrylamide gel electrophoresis showed that escherichia coli k-12 carrying the recombinant plasmids of the 2.3 kb fragment produced mba1 protein with a molecular mass of 32 kda. the recombinant plasmids carrying a 4.1 kb fragment, the other part of 6.4 kb region, produced mba2 (32 kda), mba3 (70 kda ...	1991	1795622
rapid detection of antigen binding by antibody fragments expressed in the periplasm of escherichia coli.	bacterial expression systems can greatly facilitate protein engineering of antibodies. we have developed a system for high-level expression of antibodies, antibody fragments, or hybrid antibodies with novel effector functions in the periplasm of escherichia coli. from 5 ml of cells, a simple extraction yields sufficient material for sds-gel electrophoresis, detection and characterization of hapten binding. to demonstrate our system, heavy-chain variable regions and lambda 1 light chains of a mou ...	1991	1817258
expression of honeybee prepromelittin as a fusion protein in escherichia coli.	strategies for the expression of precursors of eukaryotic secreted proteins as part of fused proteins in escherichia coli have been explored. a fusion protein with beta-galactosidase at the n-terminal end and honeybee prepromelittin at the c-terminal end (beta-gal-pm) was expressed in low amounts as a cleaved polypeptide, from which the promelittin portion had been removed. inclusion in the induction culture of 10 mm mgcl2 or 8.3% (v/v) ethanol, inhibitors of signal peptidase, gave rise to the f ...	1991	1821810
some requirements for the secretion of tumour necrosis factor and its cytotoxic activity in vitro.	the bacterial lipopolysaccharide (lps) in vitro-induced secretion of tumour necrosis factor (tnf) by mouse macrophages is described. the highest amount of tnf was produced by dba/2 mouse peritoneal macrophages activated by a dose of 1 microgram lps for 3 h. the yield of tnf was higher in the presence of indomethacin during lps induction. a markedly positive effect on tnf yield of the priming of macrophages in vivo was manifested by proliferation of peritoneal macrophages and increased secretion ...	1991	1822445
effect of il-3 and e. coli lps on the proliferation of mouse bone marrow cells in vitro.	bone marrow cells from adult balb/c mice were cultured at 37 degrees c, with 5% co2 in air, in rpmi 1640 medium complemented with fetal calf serum. the addition of il-3 (5% of wehi-3-conditioned medium) or e. coli lipopolysaccharides (lps, 50 micrograms/ml) to the cultures stimulated cell proliferation (1.29- and 1.22-fold, respectively, relative to control culture), whereas the simultaneous addition of the two factors reduced the number of cells recovered by 38% relative to those from control c ...	1991	1823004
an interleukin-1 receptor antagonist blocks lipopolysaccharide-induced colony-stimulating factor production and early endotoxin tolerance.	in this report, administration of a recombinant interleukin-1 receptor antagonist protein to mice was found to inhibit induction of colony-stimulating factor as well as induction of early endotoxin tolerance by lipopolysaccharide. these findings provide direct evidence that interleukin-1 is an intermediate in these two lipopolysaccharide-induced phenomena.	1991	1825485
production and characterization of a monoclonal antibody to the v-mos oncogene protein.	valuable information about proto-oncogenes and their physiological functions has been obtained by studying their expression in normal cells. however, the protein product of the c-mos gene, the cellular homologue of the transforming gene (v-mos) of moloney murine sarcoma virus, has not been detected in normal mouse cells or tissues. here, we have constructed a v-mos expression vector, pri-delta mos, which directs the synthesis of a truncated v-mos gene product, a protein a fusion protein. using t ...	1991	1833309
accumulation of the c-mos protein is correlated with post-natal development of skeletal muscle.	previously we reported that c-mos proto-oncogene rna was developmentally up-regulated during post-natal maturation of the rat skeletal muscle. using two different site-directed affinity-purified antipeptide antibodies we can observe that c-mos product (p43 c-mos) accumulates increasingly during post-natal development of the skeletal muscle and exhibits protein kinase activity. we find that in adult rat p43 c-mos is 10-fold higher in skeletal muscle than in ovaries, and 20- to 40-fold higher than ...	1991	1833718
analysis of n-methyl-n-nitrosourea-induced mutations in a shuttle vector plasmid propagated in mouse o6-methylguanine-dna methyltransferase-deficient cells in comparison with proficient cells.	a shuttle vector plasmid, pyz289, was constructed from the pz189 plasmid and polyoma virus dna. the plasmid contains a supf gene as a marker of mutation and can replicate in both escherichia coli and mouse cells. the pyz289 plasmids treated with n-methyl-n-nitrosourea were passed through mouse cells originating from skin tumors, which are either proficient (hl18) or deficient (hl8) in o6-methylguanine-dna methyltransferase activity, and mutations in the supf gene were analyzed. in the repair-def ...	1991	1834328
differential effects of retinoids on pokeweed mitogen induced b-cell proliferation vs immunoglobulin synthesis.	the effects of three retinoids, all-trans-retinoic acid (ra), 13-cis-retinoic acid (cra), and n-(4-hydroxyphenyl) retinamide (4-hpr) on mouse splenocyte responses to pokeweed mitogen (pwm) and escherichia coli lipopolysaccharide (lps) in vitro were evaluated. all three retinoids caused a dose dependent increase in the proliferative response to pwm. the retinoids hierarchy of efficacy based on potentiation of pwm-induced splenocyte proliferation was ra greater than cra greater than 4-hpr. 13-cis- ...	1991	1837010
bacterial translocation from the gastrointestinal tract in various mouse models for human diabetes.	bacterial translocation from the gastrointestinal (gi) tract to other internal organs was examined in multiple low-dose streptozotocin-injected (m-stz), single large-dose streptozotocin-injected (s-stz), alloxan-injected (alloxan), and non-obese diabetic (nod) mice. the incidence of bacterial translocation from the gi tract to the tested organs among diabetic mice was in the order of m-stz mice greater than s-stz mice greater than nod, alloxan, and control mice. the injections of insulin to m-st ...	1991	1839693
mutations that alter the charge of type i regulatory subunit and modify activation properties of cyclic amp-dependent protein kinase from s49 mouse lymphoma cells.	mutations in regulatory (r) subunit of camp-dependent protein kinase were analyzed from camp-resistant mutants of s49 mouse lymphoma cells by direct sequencing of amplified regions of mutant r subunit cdnas. eight distinct single base-change lesions were identified in 24 independent mutants that were hemizygous for expression of mutant r subunits with altered protein charge. cg----ta transitions predominated, but at----gc transitions and gc----ta transversions were also observed. four of five sp ...	1991	1847378
endogenous incorporation of nitric oxide from l-arginine into n-nitrosomorpholine stimulated by escherichia coli lipopolysaccharide in the rat.	the endogenous formation of nitrate in the rat, mouse and human occurs through cellular processes involving the oxidation of the guanido group of arginine. these processes proceed from arginine to nitric oxide with subsequent conversion to electrophilic nitrosating agents capable of forming carcinogenic nitrosamines. we have now demonstrated that endogenous nitrosamine formation can occur via cells stimulated in vivo by bacterial lipopolysaccharide (lps). the nitrosation of morpholine given to r ...	1991	1849054
investigation of herpes simplex virus type 1 genes encoding multiply inserted membrane proteins.	the herpes simplex virus type 1 genome contains four open reading frames (orfs) which are predicted to encode hydrophobic proteins with the potential to cross a membrane several times. the products of these genes (genes ul10, ul20, ul43 and ul53) have not previously been identified. to investigate the role of these proteins in the virus life cycle, we attempted to inactivate the genes individually by inserting the lacz gene from escherichia coli within the orfs. using this approach we have isola ...	1991	1849972
amino acid misincorporation during high-level expression of mouse epidermal growth factor in escherichia coli.	to determine whether the high-level expression of foreign proteins in escherichia coli can lead to frequent translational errors, we analyzed amino acid misincorporation in mouse epidermal growth factor (megf) produced as a trpe fusion protein. the megf dna does not encode phenylalanine and determining the phenylalanine content of the purified protein will measure missense errors. using this approach, we found an error frequency of about 1 in 40 for codons differing by a single base from those f ...	1991	1852602
novel immunomodulators with pronounced in vivo effects caused by stimulation of cytokine release.	beta-1,3-d-polyglucose derivatives protect mice against otherwise lethal bacterial infections. this protective effect has been considered to be mediated through mononuclear phagocytes. by using radioactive labelling, we localized the beta-1,3-d-polyglucose derivatized microbeads (gdm) during the period following injection. the gdm was recovered mainly in the milky spots of the omentum. in animals treated with gdm, the total white cell number was significantly increased in peritoneal fluid of mic ...	1991	1874801
leukotriene a4 hydrolase: determination of the three zinc-binding ligands by site-directed mutagenesis and zinc analysis.	three mutants of recombinant mouse leukotriene a4 (lta4) hydrolase (3.3.2.6) were produced by site-directed mutagenesis on cdna. the codons corresponding to his-295, his-299, or glu-318 were replaced by codons encoding tyrosine, tyrosine, and glutamine, respectively. the mutated cdnas were expressed in escherichia coli, and the three mutated proteins were purified to apparent homogeneity. none of these mutants contained significant amounts of zinc, as determined by atomic absorption spectrometry ...	1991	1881903
substrate analogues and divalent cations as inhibitors of glutamate decarboxylase from escherichia coli.	to examine the idea that glutamate decarboxylase from e. coli can be a convenient source for the study of the effects of compounds on gaba synthesis in the nervous system, a series of substrate analogues and divalent cations were tested as potential inhibitors of the bacterial enzyme. those analogues exhibiting inhibitor activity did so in a competitive manner. the most effective inhibitors were 3-mercaptopropionic acid, 4-bromoisophthalic acid and isophthalic acid which exhibited ki values of 0 ...	1991	1883399
bacterial ribosomal immunostimulants prime alveolar macrophages in vivo to produce interleukin 1 in vitro.	alveolar macrophages (ams) may play a key role in human respiratory immune defenses, partially by synthesizing and releasing interleukin 1 (il = 1). d53 (ribomunyl), a composite bacterial ribosomal immunostimulant, has been recognized as an efficient prevention of respiratory tract infections. in vitro, d53 enhances the il-1 production by mouse spleen adherent cells. a thymocyte proliferative response assay was used to evaluate the in vitro il-1 production by ams in healthy subjects who received ...	1991	1889248
identification of phosphorylation sites in the repetitive carboxyl-terminal domain of the mouse rna polymerase ii largest subunit.	the largest subunit of eukaryotic rna polymerase ii contains a carboxyl-terminal domain (ctd) which is comprised of repetitive heptapeptides with a consensus sequence tyr1-ser2-pro3-thr4-ser5-pro6-ser7. we demonstrate here that the mouse ctd expressed in and purified from escherichia coli can be phosphorylated in vitro by a p34cdc2/cdc28-containing ctd kinase from mouse ascites tumor cells. the product of this reaction, a phosphorylated form of the ctd, contains phosphoserine and phosphothreonin ...	1991	1899239
reduction of ribonucleotides by the obligate intracytoplasmic bacterium rickettsia prowazekii.	rickettsia prowazekii, an obligate intracellular parasitic bacterium, was shown to have a ribonucleotide reductase that would allow the rickettsiae to obtain the deoxyribonucleotides needed for dna synthesis from rickettsial ribonucleotides rather than from transport. in the presence of hydroxyurea, r. prowazekii failed to grow in mouse l929 cells or sc2 cells (a hydroxyurea-resistant cell line), which suggested that r. prowazekii contains a functional ribonucleotide reductase. this enzymatic ac ...	1991	1899861
mouse hepatocytes migrate to liver parenchyma and function indefinitely after intrasplenic transplantation.	one approach to gene therapy for hepatic diseases is to remove hepatocytes from an affected individual, genetically alter them in vitro, and reimplant them into a receptive locus. although returning hepatocytes to the liver itself would be advantageous, the feasibility of this approach has never been evaluated due to the inability to distinguish donor from host hepatocytes. to unambiguously identify transplanted hepatocytes after transplantation, and to better quantitate their number and degree ...	1991	1899924
expression of the vibrio cholerae gene encoding aldehyde dehydrogenase is under control of toxr, the cholera toxin transcriptional activator.	the toxr gene of vibrio cholerae encodes a transcriptional activator required for the expression of the cholera toxin genes (ctxab) and more than 15 other genes encoding secreted or membrane proteins. the latter group includes virulence genes involved in the biogenesis of the tcp pilus, the accessory colonization factor, and such toxr-activated genes as taga, mutations in which cause no detectable virulence defect in the suckling mouse model. to analyze the regulation of expression and the struc ...	1991	1902210
immunogenicity and protective efficacy of pertussis toxin subunit s1 produced by bacillus subtilis.	pertussis toxin (pt) subunit s1 was produced in bacillus subtilis as a secretory protein designated bacs1. bacs1 was partially purified and used to immunize mice. the sera were tested for pt-neutralizing antibodies and for protective capacity in a mouse model. unlike previous findings with recombinant s1 from escherichia coli, the recombinant bacs1 protein induced antibodies that were both neutralizing and protective. an adjuvant was necessary for efficient immunization with bacs1 but not with p ...	1991	1909767
role of bacterial hemolysin production in induction of macrophage ia expression during infection with listeria monocytogenes.	the production of a hemolytic exotoxin (hly) termed listeriolysin o (llo) is a major determinant of the virulence of the gram-positive bacterium listeria monocytogenes. as determined by lethal inoculum size, llo- strains of l. monocytogenes generally are several orders of magnitude less virulent than their llo+ counterparts. the generation of protective anti-listeria t cell immunity also has been shown to depend on the llo phenotype of the bacteria present during primary infection, although the ...	1991	1918964
identification and expression of the cdna of kin17, a zinc-finger gene located on mouse chromosome 2, encoding a new dna-binding protein.	we report the cloning of kin17 cdna, 1414 bp long with an orf of 391 residues showing a zinc finger and nuclear localization signals. by recloning the cdna into an appropriate vector, we produced kin17 protein in e. coli, purified it partially and shown that kin17 protein binds to double-stranded dna. the kin17 gene was localized by cytogenetic mapping in mouse chromosome 2, band a. genomic sequences homologous to kin17 cdna were detected also in rat and human dnas. kin17 mrna is highly expresse ...	1991	1923796