| simultaneous expression of leukocyte-type 12-lipoxygenase and reticulocyte-type 15-lipoxygenase in rabbits. | in rabbit reticulocytes an arachidonic acid 15-lipoxygenase (15-lox) is expressed at high yield. rescreening a rabbit reticulocyte cdna library for alternative 15-lox transcripts, a full length cdna which encodes a novel lipoxygenase was isolated. the predicted amino acid sequence of this enzyme shared a high degree (99%) of identity with the reticulocyte-type 15-lipoxygenase. among the six amino acid residues different in both enzymes a phe-leu exchange was detected at position 353. recently, s ... | 1998 | 9600854 |
| prostaglandin e2 alterations during sepsis are partially mediated by endotoxin-induced inhibition of prostaglandin 15-hydroxydehydrogenase. | prostaglandin e2 (pge2) is significantly elevated in the plasma of septic or injured patients and is thought to be a component of the resultant immune suppression associated with augmented rates of infection and mortality. many studies have examined the effect of burn injury and sepsis on pge2 synthesis. however, the effect of sepsis or burn injury on the expression of prostaglandin 15-hydroxydehydrogenase (pgdh), the key enzyme responsible for pge2 degradation, has not been explored. the aim of ... | 1998 | 9603077 |
| immunisation with recombinant ama-1 protects mice against infection with plasmodium chabaudi. | the plasmodium merozoite surface antigen apical membrane antigen-1 (ama-1) has previously been shown to provide partial protection to saimiri and rhesus monkeys immunised with recombinant plasmodium fragile or parasite-derived plasmodium knowlesi ama-1, respectively. in the study reported here we have used the plasmodium chabaudi/mouse model system to extend our pre-clinical assessment of an ama-1 vaccine. we describe here the expression of the full-length plasmodium chabaudi adami ama-1 and the ... | 1998 | 9607037 |
| normal levels of the vitamin d receptor and its message in psoriatic skin. | treatment with vitamin d3 analogs improves psoriasis. the vitamin d receptor (vdr) mediates most, if not all, the effects of vitamin d3. the purpose of this study was to determine the levels of the vdr mrna and vdr protein in normal and in involved and uninvolved psoriatic skin. although vdr mrna was not detected by northern analysis of human skin samples, it was readily detectable by use of the more sensitive ribonuclease protection assay. the vdr mrna levels were normal in acute guttate as wel ... | 1996 | 9627688 |
| purification, characterization, and cloning of a cytosolic aspartyl aminopeptidase. | an aminopeptidase with a preference for n-terminal aspartyl and glutamyl residues but distinct from glutamyl aminopeptidase (ec 3.4. 11.7) was purified to near homogeneity from rabbit brain cytosol. its properties were similar to an enzyme described previously (kelly, j. a., neidle, e. l., and neidle, a. (1983) j. neurochem. 40, 1727-1734). aspartyl aminopeptidase had barely detectable activity toward simple aminoacyl-naphthylamide substrates. its activity was determined with the substrate asp-a ... | 1998 | 9632644 |
| novel recombinant analogues of bovine placental lactogen. g133k and g133r provide a tool to understand the difference between the action of prolactin and growth hormone receptors. | two new analogues of bovine placental lactogen (bpl), bpl(g133k) and bpl(g133r), were expressed in escherichia coli, refolded, and purified to a native form. binding experiments, which are likely to represent the binding to site 1 only, to intact fdc-p1 cells transfected with rabbit (rb) growth hormone receptor (ghr) or with human (h) ghr, to nb2 rat lymphoma cells, or to rabbit mammary gland membranes prolactin receptor (prlr), revealed only small or no reduction in binding capacity. the comple ... | 1998 | 9632658 |
| binding of immobilized fibronectin by biliary drain isolates. | occlusion of biliary stents, as the result of bacterial adhesion and colonization onto biliary stents, still remains a major problem. biliary proteins, such as fibronectin (fn) and vitronectin (vn), have been presumed to be involved in the process of bacterial adhesion to biliary biomaterial. in the present study, fn binding by 5 strains of e. coli isolated from biliary drains or from bile was studied. all strains did not bind detectable amounts of soluble fn but bound to immobilized plasma fn. ... | 1998 | 9638875 |
| two membrane anchors of wolinella succinogenes hydrogenase and their function in fumarate and polysulfide respiration. | wolinella succinogenes can grow by anaerobic respiration with fumarate or polysulfide as the terminal electron acceptor, and h2 or formate as the electron donor. a deltahydabc mutant lacking the hydrogenase structural genes did not grow with h2 and either fumarate or polysulfide. in contrast to the wild-type strain, the mutant grown with fumarate and with formate instead of h2 did not catalyze the reduction of fumarate, polysulfide, dimethylnaphthoquinone, or benzyl viologen by h2. growth and en ... | 1998 | 9639603 |
| correlation of the expansion segments in mammalian rrna with the fine structure of the 80 s ribosome; a cryoelectron microscopic reconstruction of the rabbit reticulocyte ribosome at 21 a resolution. | samples of 80 s ribosomes from rabbit reticulocytes were subjected to electron cryomicroscopy combined with angular reconstitution. a three-dimensional reconstruction at 21 a resolution was obtained, which was compared with the corresponding (previously published) reconstruction of escherichia coli 70 s ribosomes carrying trnas at the a and p sites. in the region of the intersubunit cavity, the principal features observed in the 70 s ribosome (such as the l1 protuberance, the central protuberanc ... | 1998 | 9642046 |
| expression of a gene for a porin-like protein of the ompa family from mycobacterium tuberculosis h37rv. | an open reading frame in the genomic database of mycobacterium tuberculosis h37rv was identified as having homology with an outer membrane protein. we found that the gene specified a protein belonging to the ompa family, which includes some porins of gram-negative organisms. the gene was amplified by pcr and cloned into escherichia coli. overexpression of the gene was toxic to the host, but limited amounts could be purified from cells before growth ceased. a truncated gene devoid of the code for ... | 1998 | 9657995 |
| expression and refolding of truncated recombinant major outer membrane protein antigen (r56) of orientia tsutsugamushi and its use in enzyme-linked immunosorbent assays. | the variable 56-kda major outer membrane protein of orientia tsutsugamushi is the immunodominant antigen in human scrub typhus infections. the gene encoding this protein from karp strain was cloned into the expression vector pet11a. the recombinant protein (r56) was expressed as a truncated nonfusion protein (amino acids 80 to 456 of the open reading frame) which formed an inclusion body when expressed in escherichia coli bl21. refolded r56 was purified and compared to purified whole-cell lysate ... | 1998 | 9665960 |
| expression of equine morbillivirus (emv) matrix and fusion proteins and their evaluation as diagnostic reagents. | full-length cdna clones coding for the matrix (m) and fusion (f) proteins of equine morbillivirus (emv) were isolated by rt-pcr, and expressed in escherichia coli using two different expression systems. western blot analysis indicated that the m and f proteins, expressed either by itself or as fusion proteins with glutathione s-transferase (gst), were insoluble and degraded after expression. analysis of the degradation pattern of recombinant m protein suggested that the n-terminus of the matrix ... | 1997 | 9672592 |
| antigenicity of vp60 structural protein of rabbit haemorrhagic disease virus. | five overlapping segments of the vp60 capsid protein gene of rabbit haemorrhagic disease virus have been expressed in e. coli under the control of the t7 rna polymerase. after purification, the antigenicity of these denatured protein segments has been studied by reactivity with sera from both naturally infected and vaccinated animals in western blot analysis. the amino terminus segments of the protein (comprising the first 175 amino acids) are highly reactive with the tested sera, between 10 and ... | 1997 | 9672641 |
| purification and partial characterization of glyceraldehyde-phosphate dehydrogenase from electric organ of electrophorus electricus (l.). | the glyceraldehyde-phosphate dehydrogenase (gapdh, ec 1.2.1.12) was purified to homogeneity from electric organ of electrophorus electricus (l.) by a hydrophobic chromatography method on deacetylcolchicine-sepharose. the purification resulted in a 162 fold increase in specific activity of the gapdh and final yield was approximately 37%. the purified enzyme showed a single band in sds-page, with an apparent molecular mass of 36 kda. the purity of the colchicine-sepharose isolated material was ana ... | 1998 | 9679332 |
| secretion of scfv antibody fragments. | | 1998 | 9680640 |
| development of a magnetic microplate chemifluorimmunoassay for rapid detection of bacteria and toxin in blood. | a magnetic microplate chemifluorimmunoassay (mmcia) is described using an immunomagnetic separation and a fluorescent microplate technique for rapid detection of low-level escherichia coli o157:h7, bacillus subtilis var. niger spores, and staphylococcal enterotoxin type b from whole blood. in general, the mmcia has at least several-fold more sensitivity than the conventional enzyme-linked immunosorbent assay. in addition, the assay sensitivities using direct fluorochrome label as the reporter, o ... | 1998 | 9683505 |
| ca2+-sensitizing effects of the mutations at ile-79 and arg-92 of troponin t in hypertrophic cardiomyopathy. | several mutations in human cardiac troponin t (tnt) gene have been reported to cause hypertrophic cardiomyopathy (hcm). to explore the effects of the mutations on cardiac muscle contractile function under physiological conditions, human cardiac tnt mutants, ile79asn and arg92gln, as well as wild type, were expressed in escherichia coli and exchanged into permeabilized rabbit cardiac muscle fibers, and ca2+-activated force was determined. the free ca2+ concentrations required for tension generati ... | 1998 | 9688851 |
| effects of mk-886, a leukotriene biosynthesis inhibitor, in a rabbit model of endotoxic shock. | leukotrienes are one of the biological mediators that play a role in endotoxic shock. in this study, we investigated the effects of a leukotriene biosynthesis inhibitor, mk-886, in a rabbit model of endotoxic shock. lipopolysaccharide (escherichia coli serotype 055:b5) infusion (1 mg kg(-1) h(-1)) to rabbits caused a biphasic decline in arterial blood pressure and decreased the vasoresponsiveness to phenylephrine, potassium chloride, sodium nitroprusside and acetylcholine in abdominal aortic rin ... | 1998 | 9696411 |
| control of bacterial infections in the hard tick dermacentor variabilis (acari: ixodidae): evidence for the existence of antimicrobial proteins in tick hemolymph. | the ability of hard ticks to prevent infection by bacteria was investigated. during a 72-h period, virtually all dermacentor variabilis females survived inoculation with bacillus subtilis, escherichia coli, and staphyloccocus aureus but few survived infection with pseudomonas aeruginosa. the hemocyte population increased to peak abundance at 48 h to approximately 6 times that of the uninfected controls. in contrast, the soluble hemolymph protein content decreased inversely as the hemocytes incre ... | 1998 | 9701927 |
| antigenic structure of the capsid protein of rabbit haemorrhagic disease virus. | rabbit haemorrhagic disease virus (rhdv) causes an important disease in rabbits. the virus capsid is composed of a single 60 kda protein. the capsid protein gene was cloned in escherichia coli using the pet3 system, and the antigenic structure of rhdv vp60 was dissected using 11 monoclonal antibodies (mabs) and 12 overlapping fragments of the protein expressed in e. coli. two antigenic regions were found. ten out of the 11 mabs recognized different discontinuous epitopes in the most immunodomina ... | 1998 | 9714238 |
| serp2, an inhibitor of the interleukin-1beta-converting enzyme, is critical in the pathobiology of myxoma virus. | recently, myxoma virus was shown to encode an additional member of the serpin superfamily. the viral gene, called serp2, was cloned, and the serp2 protein was shown to specifically bind to interleukin-1beta (il-1beta)-converting enzyme (ice), thus inhibiting the cleavage of pro-il-1beta by the protease (f. petit, s. bertagnoli, j. gelfi, f. fassy, c. boucraut-baralon, and a. milon, j. virol. 70:5860-5866, 1996). here, we address the role of serp2 in the development of myxomatosis, a lethal infec ... | 1998 | 9733819 |
| identification of sigma 32-like factors and ftsx-rpoh gene arrangements in enteric bacteria. | western blot analyses using anti-escherichia coli k-12 sigma 32 antibodies and southern blot analyses using rpoh and ftsx dna probes were performed using different enteric bacteria. results show that the bacterial strains analysed have sigma 32-like transcription factors and ftsx and rpoh homologs in a similar map position. although the presence of sigma 32-like factors seems to be extended to all proteobacteria, rpoh and ftsx homologs seem to be present as neighbors in the genome only in the en ... | 1998 | 9734306 |
| anchoring antibodies to membranes using a diphtheria toxin t domain-zz fusion protein as a ph sensitive membrane anchor. | we have constructed a fusion protein, t-zz, in which the igg-fc binding protein zz was fused to the c-terminus of the diphtheria toxin transmembrane domain (t domain). while soluble at neutral ph, t-zz retained the capacity of the t domain to bind to phospholipid membranes at acidic ph. once anchored to the membrane, the zz part of the protein was capable of binding mouse monoclonal or rabbit polyclonal igg. our results show that the t-zz protein can function as a ph sensitive membrane anchor fo ... | 1998 | 9738938 |
| immunodiagnosis of clonorchiasis using a recombinant antigen. | a cdna expression library of clonorchis sinensis adult worm was constructed, and screened out immunologically. one clone, pbcs31, was selected in view of its predominant reactivity with an experimentally infected rabbit serum. recombinant c. sinensis antigen with 28 kda as a beta-galactosidase fusion protein produced in escherichia coli was identified by immunoblot analysis. the cloned gene was composed of 16 copies of a 30 base pair repeat and an additional 320 bases. the deduced amino acid seq ... | 1998 | 9755589 |
| partial liquid ventilation reduces pulmonary neutrophil accumulation in an experimental model of systemic endotoxemia and acute lung injury. | to determine whether pulmonary neutrophil sequestration and lung injury are affected by partial liquid ventilation with perfluorocarbon in a model of acute lung injury (ali). | 1998 | 9781729 |
| [the corrective action of antibodies in experimental intestinal dysbacteriosis]. | as shown in this work, antisera obtained after the immunization of animals with vaccines, prepared from salmonella minnesota strain r595 (re mutant) or escherichia coli o14 having enterobacterial common antigen (eca), as well as human antisera with elevated titers of antibodies to re glycolipid or to lps o14 (eca), inhibited the development of experimental intestinal dysbacteriosis in white mice, induced by the administration of ampyox in large doses. the degree of the inhibiting action of the a ... | 1998 | 9783390 |
| identification of 30 kda calsequestrin-binding protein, which regulates calcium release from sarcoplasmic reticulum of rabbit skeletal muscle. | in a previous study [yamaguchi, kawasaki and kasai (1995) biochem. biophys. res. commun. 210, 648-653], we showed that the stilbene derivative 4,4'-di-isothiocyanostilbene-2,2'-disulphonic acid activates the ca2+ channel in the sarcoplasmic reticulum (sr) in rabbit skeletal muscle, and it does not bind to the channel protein itself but to the sr 30 kda protein. furthermore, the 30 kda protein was shown to bind to calsequestrin (csq), which is one of the regulators of the ca2+ release channel in ... | 1998 | 9794793 |
| serological analysis of c-terminal region of alpha antigen from mycobacterium avium-intracellulare complex and mycobacterium tuberculosis. | the alpha antigen, which is a 30 kda protein secreted by mycobacterial species, is an immunodominant antigen. the c-terminal regions of alpha antigens are highly divergent, though there are regions where the amino acid sequence of alpha antigen is conserved. we investigated whether the c-terminal regions of the mycobacterium avium alpha antigen, m. intracellulare alpha antigen and m. tuberculosis alpha antigen contain sequence-specific b-cell epitopes. the c-terminal regions of m. avium alpha an ... | 1998 | 9808416 |
| western blotting analysis of heat shock proteins of rickettsiales and other eubacteria. | heat shock proteins (hsp) of four rickettsia species, three bartonella species, two ehrlichia species, orientia tsutsugamushi and seventeen other eubacterial species were characterized by the enhanced chemiluminescence western blotting (wb) technique with antibodies raised against recombinant hsp from escherichia coli and purified groes from r. typhi. although e. coli dnak and groel have epitopes that are highly conserved among the homologous proteins found in rickettsia, ehrlichia, o. tsutsugam ... | 1998 | 9809424 |
| function and protective capacity of treponema pallidum subsp. pallidum glycerophosphodiester phosphodiesterase. | infectious syphilis, caused by the spirochete bacterium treponema pallidum subsp. pallidum, remains a public health concern worldwide. the immune-response evasion mechanisms employed by t. pallidum are poorly understood, and prior attempts to identify immunoprotective antigens for subsequent vaccine design have been unsuccessful. previous investigations conducted in our laboratory identified the t. pallidum glycerophosphodiester phosphodiesterase as a potential immunoprotective antigen by using ... | 1998 | 9826352 |
| at least three linear regions but not the zinc-finger domain of u1c protein are exposed at the surface of the protein in solution and on the human spliceosomal u1 snrnp particle. | no structural information on u1c protein either in its free state or bound to the spliceosomal u1 small nuclear ribonucleoprotein (snrnp) particle is currently available. using rabbit antibodies raised against a complete set of 15 u1c overlapping synthetic peptides (16-30 residues long) in different immunochemical tests, linear regions exposed at the surface of free and u1 snrnp-bound u1c were identified. epitopes within at least three regions spanning residues 31-62, 85-103 and 116-159 were rec ... | 1998 | 9826776 |
| structural determination of the o-antigenic polysaccharide from escherichia coli o35 and cross-reactivity to salmonella arizonae o62. | the structure of the o-antigenic polysaccharide from escherichia coli o35 has been investigated with the aid of nmr spectroscopy, sugar and methylation analyses. the sequence of the sugar residues could be determined by noesy and heteronuclear-multiple-bond-connectivity nmr experiments. the polysaccharide is composed of hexasaccharide repeating units with the following structure, where rha and galnacan represent rhamnose and 2-acetamido-2-deoxy-galacturonamide, respectively: carbohydrate sequenc ... | 1998 | 9851702 |
| ex vivo gene transfer of endothelial nitric oxide synthase to atherosclerotic rabbit aortic rings improves relaxations to acetylcholine. | cholesterol feeding results in impaired endothelium dependent vasorelaxation. the role of nitric oxide in this process is unclear. the aim of this study was to evaluate the role of nitric oxide in cholesterol-induced vasomotor dysfunction by examining the effect of overexpression of enos in the hypercholesterolemic rabbit aorta on vascular reactivity. vascular rings from the thoracic aorta of hypercholesterolemic rabbits were exposed ex vivo either to an adenoviral vector encoding endothelial ni ... | 1998 | 9862175 |
| [estimation of probiotics by infection model of infant rabbit with enterohemorrhagic escherichia coli o157:h7]. | we examined the preventative effect against enterohemorrhagic escherichia coli o157:h7 (ehec) infection in rabbits on administration of probiotics contained in streptococcus faecalis, clostridium butyricum and bacillus mesentericus. the probiotics were administered to 5 days old japanese white rabbits throughout the experiment. inoculation of 10(6) colony forming units (cfus) of ehec per rabbit was tried 8 days after birth. three days after ehec inoculation, in the control group (not administere ... | 1998 | 9916417 |
| synthetic peptides corresponding to the beta-hairpin loop of rabbit defensin np-2 show antimicrobial activity. | mammalian defensins, a class of antibacterial peptides, are composed of 29-35 amino acids with six cysteines which form three disulfide bonds. structural studies indicate a triple stranded beta-sheet structure with a well defined beta-hairpin loop at the c-terminal region. it is demonstrated in this report that 18 and 26 residue synthetic peptides corresponding to the beta-hairpin region, constrained by a single disulfide bond, have potent antimicrobial activity without hemolytic activity. circu ... | 1999 | 9918829 |
| immunoglobulin m-enriched human intravenous immunoglobulin prevents complement activation in vitro and in vivo in a rat model of acute inflammation. | an important antiinflammatory mechanism of intravenous immunoglobulin preparations (ivig) is their ability to block complement activation. the purpose of this study was to compare the complement-inhibitory activity of four ivig preparations differing in isotype composition. the preparations were: (1) ivigg (48 g/l igg, 2 g/l iga; intraglobin f); (2) pentaglobin (38 g/l igg, 6 g/l igm, 6 g/l iga); (3) ivigm (35 g/l igm, 12 g/l iga, 3 g/l igg); and (4) iviga (41 g/l iga, 9 g/l igg), all from biote ... | 1999 | 9920844 |
| the nucleotide sequence of the 3'-terminal region of dasheen mosaic virus (caladium isolate) and expression of its coat protein in escherichia coli for antiserum production. | a caladium isolate of dasheen mosaic virus (dsmv-ch) was cloned as cdna from genomic rna. the sequence of the 3'-terminal 3158 nucleotides, which consisted of the 3'-terminus of the nia gene, the nib gene, the coat protein (cp) gene, and a 246-nucleotide non-coding region, was between 57-68% similar at the nucleotide level and 72-82% similar at the amino acid level when compared with other potyviruses. phylogenetic analysis of aligned, selected potyviral cp sequences indicate that dsmv-ch is sim ... | 1998 | 9930202 |
| immunogenicity of actinobacillus apxia toxin epitopes fused to the e. coli heat-labile enterotoxin b subunit. | peptides kdygastgssl (epil). sllrrrrngedvsv (epi3) and ddeiygndghp (epi6), predicted to constitute immunogenic epitopes of the hemolysin-cytotoxin apxia of actinobacillus pleuropneumoniae were inserted into a surface-exposed loop of the b subunit of the e. coli heat-labile enterotoxin (etxb). the resulting chimeric proteins were recognized by monospecific antibodies against purified native apxi and by convalescent sera of pigs that were positive for a. pleuropneumoniae serotype 1. mice anti-sera ... | 1999 | 10073721 |
| construction and characterization of an equine herpesvirus 1 glycoprotein c negative mutant. | an equine herpesvirus 1 (ehv-1) strain racl 11 mutant was constructed that carries the escherichia coli lacz gene instead of the open reading frame encoding glycoprotein c (gc). the engineered virus mutant (l11(delta)gc) lacked codons 46-440 of the 1404 bp gene. on rabbit kidney cell line rk13 and equine dermal cell line edmin337, the l11(delta)gc virus grew to titers which were reduced by approximately 5- to 10-fold compared with wild-type racl11 virus or a repaired virus (r-l11(delta)gc). howe ... | 1999 | 10082388 |
| ca2+ sensitization and potentiation of the maximum level of myofibrillar atpase activity caused by mutations of troponin t found in familial hypertrophic cardiomyopathy. | human wild-type cardiac troponin t, i, c and five troponin t mutants (i79n, r92q, f110i, e244d, and r278c) causing familial hypertrophic cardiomyopathy were expressed in escherichia coli, and then were purified and incorporated into rabbit cardiac myofibrils using a troponin exchange technique. the ca2+-sensitive atpase activity of these myofibrillar preparations was measured in order to examine the functional consequences of these troponin mutations. an i79n troponin t mutation was found to cau ... | 1999 | 10085122 |
| synergistic effect of hyperoxia and immunoglobulin a on mucosal barrier defense. | hyperoxia has been reported to be protective against gut-derived sepsis. although secretory immunoglobulin a (iga) is primarily responsible for humoral defense of mucosal surfaces, a potential synergistic effect with hyperoxia is unknown. an asanguineous cell monolayer system was used to study these aspects in vitro. | 1999 | 10088836 |
| up-regulation of microsphere transport across the follicle-associated epithelium of peyer's patch by exposure to streptococcus pneumoniae r36a. | transport of antigens through the follicle-associated epithelium (fae) of peyer's patch (pp) is the critical first step in the induction of mucosal immune responses. we have previously described that short-term exposure to streptococcus pneumoniae r36a induced dramatic morphological alterations of the fae in rabbit pp. these results prompted us to investigate whether the pneumococci-induced modifications were accompanied by enhanced ability of the fae to transport antigens. we addressed this pro ... | 1999 | 10094922 |
| characterization of the cardiac holotroponin complex reconstituted from native cardiac troponin t and recombinant i and c. | cardiac troponin i (ctni), the inhibitory subunit of cardiac troponin (ctn), is phosphorylated by the camp-dependent protein kinase a at two adjacently located serine residues within the heart-specific n-terminal elongation. four different phosphorylation states can be formed. to investigate each monophosphorylated form ctni mutants, in which each of the two serine residues is replaced by an alanine, were generated. these mutants, as well as the wild-type cardiac troponin i (ctni-wt) have been e ... | 1999 | 10103031 |
| essential role of helicobacter pylori gamma-glutamyltranspeptidase for the colonization of the gastric mucosa of mice. | constitutive expression of gamma-glutamyltranspeptidase (ggt) activity is common to all helicobacter pylori strains, and is used as a marker for identifying h. pylori isolates. helicobacter pylori ggt was purified from sonicated extracts of h. pylori strain 85p by anion exchange chromatography. the n-terminal amino acid sequences of two of the generated endo-proteolysed peptides were determined, allowing the cloning and sequencing of the corresponding gene from a genomic h. pylori library. the h ... | 1999 | 10200957 |
| the crystal structure of the escherichia coli maltodextrin phosphorylase-acarbose complex. | acarbose is a naturally occurring pseudo-tetrasaccharide. it has been used in conjunction with other drugs in the treatment of diabetes where it acts as an inhibitor of intestinal glucosidases. to probe the interactions of acarbose with other carbohydrate recognition enzymes, the crystal structure of e. coli maltodextrin phosphorylase (malp) complexed with acarbose has been determined at 2.95 a resolution and refined to crystallographic r-values of r (rfree) = 0.241 (0.293), respectively. acarbo ... | 1999 | 10220320 |
| ultrasonic enhancement of antibiotic action on escherichia coli biofilms: an in vivo model. | biofilm infections are a common complication of prosthetic devices in humans. previous in vitro research has determined that low-frequency ultrasound combined with aminoglycoside antibiotics is an effective method of killing biofilms. we report the development of an in vivo model to determine if ultrasound enhances antibiotic action. two 24-h-old escherichia coli (atcc 10798) biofilms grown on polyethylene disks were implanted subcutaneously on the backs of new zealand white female rabbits, one ... | 1999 | 10223938 |
| effects of endothelin-1 on bacterial clearance in rabbits. | as elevated endothelin-1 (et-1) levels have been reported in systemic inflammatory diseases, the role of et-1 as a promoter of inflammatory reactions is currently under investigation. the purpose of this study was to investigate the potential influence of et-1 on systemic vascular pressure and immune function in terms of blood clearance and organ distribution of injected escherichia coli in a rabbit model. to enable quantification of the clearance process, defined numbers of exogenous e. coli (1 ... | 1999 | 10225166 |
| the effect of the lipid a analog, e5531 on fever induced by endotoxin from escherichia coli. | e5531 is a specific lipid a antagonist and is expected to be a drug for the treatment of septic shock. the lal (limulus amebocyte lysate) activity and pyrogenicity of e5531 were determined. the lal activity of e5531 is large and confirmed that e5531 had a high affinity to the lipopolysaccharide receptor. the pyrogenic activity of e5531 is weak and the pyrogenic profile of e5531 is different from that of usp (united states pharmacopoeia) reference standard endotoxin (etx). e5531 suppressed the py ... | 1999 | 10328569 |
| antigenic profile of african horse sickness virus serotype 4 vp5 and identification of a neutralizing epitope shared with bluetongue virus and epizootic hemorrhagic disease virus. | african horse sickness virus (ahsv) causes a fatal disease in horses. the virus capsid is composed of a double protein layer, the outermost of which is formed by two proteins: vp2 and vp5. vp2 is known to determine the serotype of the virus and to contain the neutralizing epitopes. the biological function of vp5, the other component of the capsid, is unknown. in this report, ahsv vp5, expressed in insect cells alone or together with vp2, was able to induce ahsv-specific neutralizing antibodies. ... | 1999 | 10329555 |
| biliary pancreatic reflux-induced acute pancreatitis--myth or possibility? | the mechanism whereby gallstone passage through the choledochoduodenal junction initiates acute pancreatitis is not known. we mimicked different patterns of stone impaction at the choledochoduodenal junction in a rabbit model and studied whether these result in biliary pancreatic reflux and the initiation of pancreatic inflammation. | 1999 | 10333208 |
| prevention of lethal acute hepatic failure by antimacrophage migration inhibitory factor antibody in mice treated with bacille calmette-guerin and lipopolysaccharide. | during the past few years, the biological functions of macrophage migration inhibitory factor (mif) have been extensively re-evaluated. this has been found to be protein involved in broad-spectrum pathophysiological states as an inflammatory cytokine, pituitary-derived hormone, and glucocorticoid-induced immunomodulator. in this study, we investigated the involvement of this cytokine in the pathogenesis of lethal liver injury. injecting a small dose of lipopolysaccharide (lps) into bacille calme ... | 1999 | 10347118 |
| isolation and purification of a campylobacter upsaliensis autolysin. | autolytic activity in the soluble and sediment fractions of sonicates of the spiral and the coccoid form of campylobacter upsaliensis could not be demonstrated by native (nondenaturing) polyacrylamide gel electrophoresis (page). autolysins were detected, however, by using denaturing sodium dodecyl sulfate (sds)-page gels containing either purified escherichia coli peptidoglycan or whole cells of micrococcus luteus (micrococcus lysodeikticus) as the turbid substrate, with subsequent renaturation ... | 1999 | 10349717 |
| comparison of escherichia coli and rabbit reticulocyte ribosome display systems. | ribosome display is a technology for library selection and simultaneous molecular evolution in vitro. we present here a comparison between an optimized escherichia coli system and different rabbit reticulocyte ribosome display systems, optimized in a number of parameters, as a coupled eukaryotic system had been suggested to result in high enrichment factors [he and taussig (1997) nucleic acids res. 25, 5132-5134]. with all systems, antibody scfv fragments, complexed to the ribosomes and the corr ... | 1999 | 10350066 |
| efficacy of beta-lactam and inhibitor combinations in a diffusion chamber model in rabbits. | using a diffusion chamber in rabbits, we evaluated therapy with the combination of ceftriaxone plus the beta-lactamase inhibitor tazobactam in comparison with ceftriaxone alone. one sensitive and one resistant strain of escherichia coli, enterobacter cloacae and klebsiella pneumoniae were inoculated into one of the six diffusion chambers, implanted in the same animal. in order to simulate pharmacokinetics in humans, both substances were administered in decreasing doses. ceftriaxone was given 0, ... | 1999 | 10350378 |
| videomicroscopy provides accurate in vivo assessment of pulmonary microvascular reactivity in rabbits. | when defining the mechanism of hypoxic pulmonary vasoconstriction (hpv), investigators have employed ex vivo preparations because of the belief that accurate, quantitative assessment of pulmonary microvessels could not be obtained in vivo. we hypothesize that accurate, quantitative assessment of pulmonary microvascular reactivity can be performed using a simple, in vivo preparation. our aim was to provide this quantitative assessment in a defined animal model, and to confirm that the chosen prep ... | 1999 | 10353544 |
| the antiplatelet activity of escherichia coli lipopolysaccharide is mediated through a nitric oxide/cyclic gmp pathway. | in this study, escherichia coli lps dose-dependently (100-500 microg/ml) and time-dependently (10-60 min) inhibited platelet aggregation in human and rabbit platelets stimulated by agonists. lps also dose-dependently inhibited the intracellular ca2+ mobilization in human platelets stimulated by collagen. in addition, lps (200 and 500 microg/ml) significantly increased the formation of cyclic gmp but not cyclic amp in platelets. lps (200 microg/ml) significantly increased the production of nitrat ... | 1999 | 10359060 |
| brainstem mechanisms of autonomic dysfunction in encephalopathy-associated shiga toxin 2 intoxication. | acute encephalopathy is the major determinant of death in an early stage of shiga toxin (stx)-producing escherichia coli infection. rapid progress toward refractory hypotension and dysfunction of breathing implies autonomic center dysfunction of patients. to clarify whether autonomic dysfunction becomes an ultimate cause of death in shiga toxemia, we injected purified stx2 (20 microg/kg) intravenously into rabbits, and monitored changes in cardiovascular and respiratory function together with re ... | 1999 | 10360763 |
| bacterial expression and functional characterization of a rat thyroid hormone sulfotransferase, st1b1. | at least three forms of phenol sulfotransferase (st) st1b1, st1a1 and st1c1 are contained in rat livers. to identify the form contributing to the metabolism of 3,3',5-triiodothyronine (t3), functional characterization of these forms was performed by expression in escherichia coli. st1b1 and st1c1 were shown to be active on sulfation towards t3 with high affinity (km: 44.4 and 25.8 microm, respectively), whereas st1a1 had low affinity. in western blotting using antibodies raised against the indiv ... | 1999 | 10361887 |
| a novel mutant, apoa-i nichinan (glu235-->0), is associated with low hdl cholesterol levels and decreased cholesterol efflux from cells. | a novel variant of apolipoprotein (apo) a-i associated with low high density lipoprotein (hdl) cholesterolemia has been identified in a japanese family during screening for apoa-i variants by isoelectric focusing (ief) gel analysis. apoa-i (glu235-->0) nichinan was caused by a 3-bp deletion of nucleotides 1998 through 2000 in exon 4 of the apoa-i gene. four subjects in the family were heterozygous carriers for this mutation; the mean plasma concentrations of apoa-i and hdl cholesterol of affecte ... | 1999 | 10364075 |
| domains of rinderpest virus phosphoprotein involved in interaction with itself and the nucleocapsid protein. | the yeast two-hybrid system was used to identify domains involved in specific in vivo interactions between the rinderpest virus (rpv) phosphoprotein (p) and nucleocapsid protein (n). n and p genes were cloned in both the yeast gal4 dna-binding and gal4 activation domain vectors, which enabled analysis of self and interprotein interactions. mapping of the domain of p protein involved in its association with itself revealed that the cooh-terminal 32 amino acids (316-347) that forms a part of the h ... | 1999 | 10366579 |
| fractionation of polyclonal antibodies to fragments of a neuroreceptor using three increasingly chaotropic solvents. | we have developed specific antibodies against fragments of anaplastic lymphoma kinase (alk) in order to develop tools for characterizing the expression and biological function of this orphan receptor. the first fragment consisted of residues 280 to 480 of the murine extracellular domain, was expressed in escherichia coli (e. coli), purified in the presence of urea from the pellet of mechanically lysed cells and injected into rabbits as an unfolded protein in urea. the second fragment consisted o ... | 1999 | 10379656 |
| distribution of two isoforms of nadp-dependent isocitrate dehydrogenase in soybean (glycine max). | two different cdnas that encode nadp-specific isocitrate dehydrogenase (nadp-idh) isozymes of soybean (glycine max) were characterized. the nucleotide sequences of the coding regions of these cdnas have 74% identity to each other and give predicted amino acid sequences that have 83% identity to each other. using pcr techniques, their coding regions were subcloned into a protein overexpression vector, pqe32, to yield pidh4 and pidh1, respectively. both idh4 and idh1 enzymes were expressed in esch ... | 1999 | 10394941 |
| growth phase-dependent regulation of vsr endonuclease may contribute to 5-methylcytosine mutational hot spots in escherichia coli. | using rabbit polyclonal antibodies, we have shown that the dcm cytosine methylase of escherichia coli is maintained at a constant level during cell growth, while vsr endonuclease levels are growth phase dependent. decreased production of vsr relative to dcm during the log phase may contribute substantially to the mutability of 5-methylcytosine. | 1999 | 10400606 |
| functional consequences of a carboxyl terminal missense mutation arg278cys in human cardiac troponin t. | a carboxyl terminal missense mutant arg278cys of human cardiac troponin t that causes familial hypertrophic cardiomyopathy was expressed in escherichia coli, purified, and exchanged into rabbit cardiac skinned muscle fibers using a troponin exchange technique. compared to the fibers exchanged with human cardiac wild-type troponin t, the fibers exchanged with the mutant arg278cys developed less maximum force with a decreased cooperativity and a slightly increased ca(2+) sensitivity, resulting in ... | 1999 | 10405326 |
| evaluation of commercially available escherichia coli j5 bacterin as protection against experimental challenge with pasteurella multocida in rabbits. | to evaluate the ability of commercially available escherichia coli j5 bacterin to protect rabbits from experimental challenge with pasteurella multocida. | 1999 | 10407479 |
| metabolism of benzo[a]pyrene to trans-7,8-dihydroxy-7, 8-dihydrobenzo[a]pyrene by recombinant human cytochrome p450 1b1 and purified liver epoxide hydrolase. | recombinant human enzymes expressed in membranes obtained from escherichia coli transformed with cytochrome p450 (p450) and nadph-p450 reductase cdnas were used to identify the human p450 enzymes that are most active in catalyzing the oxidative transformation of benzo[a]pyrene in vitro. activation of benzo[a]pyrene to genotoxic products that cause induction of umu gene expression in salmonella typhimurium nm2009 by p450 1a1 and p450 1b1 enzymes was found to be enhanced by inclusion of purified e ... | 1999 | 10409402 |
| characterization of the bacillus macerans cyclodextrin glucanotransferase overexpressed in escherichia coli. | cyclodextrin glucanotransferase (cgtase, ec 2. 4. 1. 19) converts starch and related alpha-1,4-glucans to cyclodextrin (cd). our previous studies of the enzyme have suggested that e344 on the polypeptide is crucial to the enzyme activity. mutational analysis of cgtase was performed to confirm this idea. three mutant cgtases containing either e344d, e344k or e344l substitution were overexpressed in escherichia coli. however, only the wild-type and e344d cgtases became soluble when expressed at 20 ... | 1999 | 10420654 |
| preparation and characterization of recombinant dolphin fish (coryphaena hippurus) growth hormone. | dolphin fish (coryphaena hippurus) growth hormone (dfgh) cdna encoding the mature protein was cloned in a pet11a expression vector and expressed in escherichia coli bl21 cells upon induction with isopropyl-1-thio-beta-d-galactopyranoside as an insoluble protein. the expressed protein, contained within the inclusion-body pellet, was solubilized in 4.5 m urea, refolded at ph 11.3 in the presence of catalytic amounts of cysteine, and purified to homogeneity, as evidenced by sds-page. gel filtration ... | 1999 | 10425163 |
| does bile of patients with acute gallstone pancreatitis cause pancreatic inflammatory lesions? a study of the pancreatic toxicity of choledochal secretions collected at ercp. | bile flow into the pancreatic duct has been proposed as the cause of acute biliary pancreatitis. however, the pancreatic toxicity of choledochal bile from patients with acute gallstone pancreatitis has not been studied. | 1999 | 10425414 |
| role of the c-terminal extremities of the smooth muscle myosin heavy chains: implication for assembly properties. | the two light meromyosin isoforms from rabbit smooth muscle were prepared as recombinant proteins in escherichia coli. these species which differed only by their c-terminal extremity showed the same circular dichroism spectra and endotherms in measurements of differential scanning calorimetry. their solubility properties were different at ph 7.0 in the absence of monovalent salts. their paracrystals formed at low ph differed by their aspect and number. these data suggest a role for the c-termina ... | 1999 | 10431827 |
| minicircle: an improved dna molecule for in vitro and in vivo gene transfer. | minicircles are a new form of supercoiled dna molecule for nonviral gene transfer which have neither bacterial origin of replication nor antibiotic resistance marker. they are thus smaller and potentially safer than the standard plasmids currently used in gene therapy. they were obtained in e. coli by att site-specific recombination mediated by the phage lambda integrase, which was used to excise the expression cassette from the unwanted plasmid sequences. we produced two minicircles containing ... | 1999 | 10435105 |
| dynamic measurement of myosin light-chain-domain tilt and twist in muscle contraction. | a new method is described for measuring motions of protein domains in their native environment on the physiological timescale. pairs of cysteines are introduced into the domain at sites chosen from its static structure and are crosslinked by a bifunctional rhodamine. domain orientation in a reconstituted macromolecular complex is determined by combining fluorescence polarization data from a small number of such labelled cysteine pairs. this approach bridges the gap between in vitro studies of pr ... | 1999 | 10440371 |
| molecular characterization of a human dna kinase. | human polydeoxyribonucleotide kinase is an enzyme that has the capacity to phosphorylate dna at 5'-hydroxyl termini and dephosphorylate 3'-phosphate termini and, therefore, can be considered a putative dna repair enzyme. the enzyme was purified from hela cells. amino acid sequence was obtained for several tryptic fragments by mass spectrometry. the sequences were matched through the dbest data base with an incomplete human cdna clone, which was used as a probe to retrieve the 5'-end of the cdna ... | 1999 | 10446193 |
| thermal unfolding of phosphorylating d-glyceraldehyde-3-phosphate dehydrogenase studied by differential scanning calorimetry. | thermal unfolding parameters were determined for a two-domain tetrameric enzyme, phosphorylating d-glyceraldehyde-3-phosphate dehydrogenase (gapdh), and for its isolated nad(+)-binding domain. at ph 8.0, the transition temperatures (t(max)) for the apoforms of the native bacillus stearothermophilus gapdh and the isolated domain were 78.3 degrees c and 61.9 degrees c, with calorimetric enthalpies (deltah(cal)) of 4415 and 437 kj/mol (or 30.7 and 22.1 j/g), respectively. in the presence of nearly ... | 1999 | 10446379 |
| characterization of acholeplasma laidlawii ftsz gene and its gene product. | the ftsz gene was found among representatives of all bacterial groups. ftsz protein is an essential component of cell division ring. contraction of this cytoskeleton-like ring is believed to be the universal way of bacterial division. acholeplasma laidlawii possesses all features of the minimal mycoplasma cell and some traits of cell-wall bacteria and seems to be a promising object for study of basic principles of the bacterial division process. we cloned an a. laidlawii chromosomal fragment con ... | 1999 | 10448065 |
| facile characterization of translation initiation via nonsense codon suppression. | a new strategy for studying the mechanism of translation initiation in eukaryotes has been developed. the strategy involves the use of an in vitro translation system to incorporate a non-natural fluorescent amino acid into a protein from a suppressor trnaphecua misacylated with that amino acid. it is thereby possible to monitor translation initiation efficiency at an aug codon in different contexts; this is illustrated for three constructs encoding escherichia coli dihydrofolate reductase mrna w ... | 1999 | 10454635 |
| macrophage tnf mrna expression induced by lps is regulated by sphingomyelin metabolites. | metabolism of macrophage (mo) membrane phospholipids produces key mediators of inflammation and major second messengers that modulate inflammatory responses during sepsis. sphingomyelin is a major class of phospholipid that releases ceramide and sphingosine. this study was designed to investigate the involvement of sphingomyelin metabolites in mo activation by lipopolysaccharide (lps). rabbit alveolar mo were obtained by bronchoalveolar lavage and exposed to c6-ceramide, a cell-permeable analogu ... | 1999 | 10454830 |
| cloning and molecular characterization of plasmid-encoded antigens of borrelia burgdorferi. | thirteen independent clones that encode borrelia burgdorferi antigens utilizing antiserum from infection-immune rabbits were identified. the serum was adsorbed against noninfectious b. burgdorferi b31 to enrich for antibodies directed against either infection-associated antigens of b. burgdorferi b31 or proteins preferentially expressed during mammalian infection. the adsorption efficiency of the immune rabbit serum (irs) was assessed by western immunoblot analysis with protein lysates derived f ... | 1999 | 10456881 |
| induction of cellular immunosuppression by the human papillomavirus type 16 e7 oncogenic protein. | the human papillomavirus type 16 (hpv-16) e7 oncogenic protein is found in the culture supernatant of siha cells, a cervical carcinoma cell line. extracellular e7 protein, acting as a viral toxin in human immune cells, induces the overproduction of the immune suppressive ifn alpha cytokine by apcs, and inhibits the t-cell response to recall and allogenic antigens. these effects should be taken into account for the design of anti-human cervical carcinoma vaccines. | 1999 | 10472433 |
| a reverse-sandwich enzyme-linked immunosorbent assay for verocytotoxin 1 and 2 antibodies in human and bovine sera. | a reverse-sandwich enzyme-linked immunosorbent assay (elisa), in which an antibody is sandwiched by antigens, was established for the titration of antibodies to verocytotoxins (vt) in human and animal sera. this assay has two advantages over a conventional indirect elisa: (i) higher specificity and sensitivity and (ii) the ability to comparably titrate antibodies from different species. the vt1 (shiga-like toxin 1) antibody-positive rates were 5% in 202 normal adult humans and 99% in 93 normal c ... | 1999 | 10473521 |
| rp463: a stabilized technetium-99m complex of a hydrazino nicotinamide derivatized chemotactic peptide for infection imaging. | a hynic-conjugated chemotactic peptide (fmlfk-hynic) was labeled with (99m)tc using tricine and tppts as coligands. the combination of fmlfk-hynic, tricine, and tppts with (99m)tc produced a ternary ligand complex [(99m)tc(fmlfk-hynic)(tricine)(tppts)] (rp463). rp463 was synthesized either in two steps, in which the binary ligand complex [(99m)tc(fmlfk-hynic)(tricine)(2)] (rp469) was formed first and then reacted with tppts, or in one step by direct reduction of [(99m)tc]pertechnetate with stann ... | 1999 | 10502357 |
| the identification of a second cofilin binding site on actin suggests a novel, intercalated arrangement of f-actin binding. | the cofilins are members of a protein family that binds monomeric and filamentous actin, severs actin filaments, and increases monomer off-rate from the pointed end. here, we characterize the cofilin-actin interface. we confirm earlier work suggesting the importance of the lower region of subdomain 1 encompassing the n and c termini (site 1) in cofilin binding. in addition, we report the discovery of a new cofilin binding site (site 2) from residues 112-125 that form a helix toward the upper, re ... | 1999 | 10506133 |
| prevalence of virulence factors of escherichia coli strains isolated from diarrheic and healthy piglets after weaning. | this study determined the prevalence of f4, f5, f6, f17 and f41 fimbriae and the genes for feda (f18 fimbriae), lt and st enterotoxins, and shiga toxins stx1, stx2 and stx2e among e. coli isolated from 372 weaned pigs with diarrhea and 46 healthy pigs of the same age. agglutination tests showed that most isolates were negative for all five fimbrial antigens. the f4 antigen was found in 71 (19.1%) and the f5, f6, or f41 antigen was detected in 6.4% of isolates from diseased pigs. genes for the f1 ... | 1999 | 10510040 |
| adenovirus-mediated suicide gene transduction: feasibility in lens epithelium and in prevention of posterior capsule opacification in rabbits. | the most common complication of cataract surgery is the development of posterior capsule opacification (pco). hyperplasia of the lens epithelium is one of the main cellular events following phacoemulsification, and has been found to be an important feature contributing to opacification of the posterior capsule. adenoviral vector-mediated transfer is a suitable method for transducing the herpes simplex virus thymidine kinase gene (hsv-tk) into proliferating cells, allowing for the selective killi ... | 1999 | 10515456 |
| a role for the phobr regulatory system homologue in the vibrio cholerae phosphate-limitation response and intestinal colonization. | to survive and multiply in different environments, vibrio cholerae has to coordinately regulate the expression of genes involved in adaptive responses. in many pathogens, adaptive responses, including pathogenic responses, are regulated by two-component regulator (tcr) systems. it is likely that members of a tcr family play a role in the regulation of processes involved in intestinal colonization, and therefore pathogenesis, in v. cholerae. we have identified and characterized a tcr system of v. ... | 1999 | 10517599 |
| molecular cloning and characterization of a birch pollen minor allergen, bet v 5, belonging to a family of isoflavone reductase-related proteins. | birch pollen is a major cause of pollinosis and is responsible for cross-reactive oral allergies to fruits, nuts, and vegetables. the major allergen, bet v 1, has been extensively characterized, and 3 minor allergens, bet v 2, bet v 3, and bet v 4, have been cloned and sequenced. recently, another birch pollen protein with an apparent mass of 35 kd was described as a new ige-binding protein in birch pollen with cross-reacting homologues in plant foods. | 1999 | 10550744 |
| autoantibodies to brain components and antibodies to acinetobacter calcoaceticus are present in bovine spongiform encephalopathy. | bovine spongiform encephalopathy (bse) is a neurological disorder, predominantly of british cattle, which belongs to the group of transmissible spongiform encephalopathies together with creutzfeldt-jakob disease (cjd), kuru, and scrapie. autoantibodies to brain neurofilaments have been previously described in patients with cjd and kuru and in sheep affected by scrapie. spongiform-like changes have also been observed in chronic experimental allergic encephalomyelitis, at least in rabbits and guin ... | 1999 | 10569779 |
| molecular cloning, characterization, and expression of a novel human neutral sphingomyelinase. | neutral sphingomyelinase (n-smase) has emerged as an important cell membrane-associated enzyme that participates in several signal transduction and cell regulatory phenomena. using expression cloning, we have identified a 3.7-kilobase pair cdna transcript for n-smase whose open reading frame predicts a 397-amino acid polypeptide. transfection of cos-7 cells with cdna for n-smase resulted in a marked increase in n-smase activity. recombinant n-smase (r-n-smase) had the following physical-chemical ... | 1999 | 10601312 |
| adeno-associated virus type 2 nonstructural protein rep78 suppresses translation in vitro. | adeno-associated virus type 2 nonstructural protein rep78 [621 amino acids (aa) long] affects the expression of various cellular and viral genes. in this study we examined the effects of rep78 on expression of the luciferase gene from the human cytomegalovirus immediate-early promoter in hela cells and on translation of rna encoding luciferase in rabbit reticulocyte lysate. when rep78 and luciferase were coexpressed, the luciferase activity decreased despite increased levels of luciferase mrna i ... | 2000 | 10612674 |
| translocation of group 1 capsular polysaccharide to the surface of escherichia coli requires a multimeric complex in the outer membrane. | surface expression of the group 1 k30 capsular polysaccharide of escherichia coli strain e69 (o9a:k30) requires wza(k30), a member of the outer membrane auxiliary (oma) protein family. a mutation in wza(k30) severely restricts the formation of the k30 capsular structure on the cell surface, but does not interfere with the biosynthesis or polymerization of the k30 repeat unit. here we show that wza(k30) is a surface-exposed outer membrane lipoprotein. wza(k30) multimers form ring-like structures ... | 2000 | 10619844 |
| the risk of nosocomial pneumonia is not increased during partial liquid ventilation. | to determine whether partial liquid ventilation (plv) affects the risk of nosocomial pneumonia. | 1999 | 10628620 |
| a small heat shock protein cooperates with heat shock protein 70 systems to reactivate a heat-denatured protein. | small heat shock proteins (shsps) are a diverse group of heat-induced proteins that are conserved in prokaryotes and eukaryotes and are especially abundant in plants. recent in vitro data indicate that shsps act as molecular chaperones to prevent thermal aggregation of proteins by binding non-native intermediates, which can then be refolded in an atp-dependent fashion by other chaperones. we used heat-denatured firefly luciferase (luc) bound to pea (pisum sativum) hsp18.1 as a model to define th ... | 2000 | 10631262 |
| the n-terminal region of the heme-regulated eif2alpha kinase is an autonomous heme binding domain. | the n-terminal domain (ntd) of the heme-regulated eukaryotic initiation factor (eif)2alpha kinase (hri) was aligned to sequences in the ncbi data base using entrez and a pam250 matrix. significant similarity was found between amino acids 11-118 in the ntd of rabbit hri and amino acids 16-120 in mammalian alpha-globins. several conserved amino acid residues present in globins are conserved in the ntd of hri. his83 of hri was predicted to be equivalent to the proximal heme ligand (hisf8) that is c ... | 2000 | 10632719 |
| cloning and expression of a cell wall associated protein gene of mycobacterium tuberculosis. | genomic library of mycobacterium tuberculosis (sihv) was constructed into t7 promoter based vector pbluescript ii ks (+/-) in escherichia coil. the expression of mycobacterial antigens from recombinants was detected by immunoscreening using e-coli-preabsorbed hyperimmune rabbit anti-m tuberculosis cell wall associated proteins serum. among 980 clones tested by immunoscreening, a clone pjj5 was found to produce mycobacterial antigen. this clone was further characterized by immunoblotting and foun ... | 1999 | 10639772 |
| characterization of the african swine fever virus protein p49: a new late structural polypeptide. | the open reading frame b438l, located within the ecori b fragment of the african swine fever virus genome, is predicted to encode a protein of 438 amino acids with a molecular mass of 49.3 kda. it presents a cell attachment rgd (arg-gly-asp) motif but no other significant similarity to protein sequences in databases. northern blot and primer extension analysis showed that b438l is transcribed only at late times during virus infection. the b438l gene product has been expressed in escherichia coli ... | 2000 | 10640542 |
| engineering microsomal cytochrome p450 2c5 to be a soluble, monomeric enzyme. mutations that alter aggregation, phospholipid dependence of catalysis, and membrane binding. | deletion of the n-terminal membrane-spanning domain from microsomal p450s 2c5 and 2c3 generates the enzymes, 2c5dh and 2c3dh, that exhibit a salt-dependent association with membranes indicating that they retain a monofacial membrane interaction domain. the two proteins are tetramers and dimers, respectively, in high salt buffers, and only 2c5dh requires phospholipids to reconstitute fully the catalytic activity of the enzyme. amino acid residues derived from p450 2c3dh between residues 201 and 2 ... | 2000 | 10644712 |
| engineered salt-insensitive alpha-defensins with end-to-end circularized structures. | we designed a retro-isomer and seven circularized "beta-tile" peptide analogs of a typical rabbit alpha-defensin, np-1. the analogs retained defensin-like architecture after the characteristic end-to-end, cys(3,31) (c i:c vi), alpha-defensin disulfide bond was replaced by a backbone peptide bond. the retro-isomer of np-1 was as active as the parent compound, suggesting that overall topology and amphipathicity governed its antimicrobial activity. a beta-tile design with or without a single cross- ... | 2000 | 10660548 |
| [western blotting of rstv gene products in rice and insects]. | the ns3 and nc protein genes encoded by rna3 of rstv, the ncp and nsvc4 protein genes encoded by rna4 were subcloned into the e. coli expression vector pgex3x to express four groups of fusion protein under iptg induction. these fusion proteins were used to immunize rabbits to raise antisera. the antisera against the e. coli-expressed proteins were available for probing the presence of the viral gene products in both rice plant and insect hosts. the expected gene products can be probed only in di ... | 1999 | 10665229 |
| purification and characterization of chitin-binding proteins from the hemolymph of sweet potato hornworm, agrius convolvuli. | three chitin-binding proteins (cbps: cbp9, cbp15, cbp66) were identified from the larval hemolymph of sweet potato hornworm, agrius convolvuli. two (cbp9 and cbp15) of them have been isolated and purified by gel filtration (superdex hr 75), cation-exchange chromatography (mono s), and reverse-phase chromatography (murpc pc 2.1/3). in experiments to detect cbps in hemolymph, we examined whether ionic strength and existence of bovine serum albumin in the incubation solution influenced binding affi ... | 1999 | 10665375 |
| partial liquid ventilation with perflubron attenuates in vivo oxidative damage to proteins and lipids. | to determine the impact of partial liquid ventilation on the degree of pulmonary damage by reactive oxygen species in a model of acute lung injury caused by systemic endotoxemia. | 2000 | 10667523 |