Br J Rheumatol, 1995 Dec, 34(12), 1112 - 6 Presence of bacterial flora-derived antigen in synovial tissue macrophages and dendritic cells; Melief MJ et al.; In previous studies using and animal model human bacterial flora-derived peptidoglycan Polysaccharides were shown to be arthropathic after a single subcutaneous injection . A prerequisite for proof of the hypothesis that bacterial products from the normal resident flora are involved in the immune reaction of human chronic polyarthritis of unknown aetiology is the presence of these antigens in synovial tissue . 2E9, a monoclonal antibody we developed against intestinal peptidoglycan polysaccharides was used in a histochemical study in rats and stained macrophages in the spleen red pulp . In this study human synovial tissues from 10 rheumatoid arthritis (RA) and 20 non-RA patients were stained with 2E9 . We found that eight out of 10 RA patients had 2E9-positive macrophages and dendritic cells in their synovia . A significant difference was observed with the control group in which seven out of 20 were positive . No positive cells or staining of the matrix was found in the cartilage of six RA patients . These results show that exogenous bacterial antigens are present in synovial tissue macrophages and dendritic cells . It was concluded that the unknown antigen in the immune reaction in RA is not necessarily endogenous. Free Radic Biol Med, 1995 Dec, 19(6), 903 - 9 Melatonin reduces both basal and bacterial lipopolysaccharide-induced lipid peroxidation in vitro; Sewerynek E et al.; The protective effect of melatonin against lipopolysaccharide (LPS)-induced oxidative damage was examined in vitro . Lung, liver, and brain malonaldehyde (MDA) plus 4-hydroxyalkenals (4-HDA) concentrations were measured as indices of induced membrane peroxidative damage . Homogenates of brain, lung, and liver were incubated with LPS at concentrations of either 1, 10, 50, 200, or 400 micrograms/ml for 1 h and, in another study, LPS at a concentration of 400 micrograms/ml for either 0, 15, 30, or 60 min . Melatonin at increasing concentrations from 0.01-3 mM either alone or together with LPS (400 micrograms/ml) was used . Liver, brain, and lung MDA + 4-HDA levels increased after LPS at concentrations of 10, 50, 200 or 400 micrograms/ml; this effect was concentration-dependent . The highest levels of lipid peroxidation products were observed after tissues were incubated with an LPS concentration of 400 micrograms/ml for 60 min; in liver and lung this effect was totally suppressed by melatonin and partially suppressed in brain in a concentration-dependent manner . In addition, melatonin alone was effective in brain at concentrations of 0.1 to 3 mM, in lung at 2 to 3 mM, and in liver at 0.1 to 3 mM; in all cases, the inhibitory effects of melatonin on lipid peroxidation were always directly correlated with the concentration of melatonin in the medium . The results show that the direct effect of LPS on the lipid peroxidation following endotoxin exposure is markedly reduced by melatonin. J Neuroimmunol, 1995 Dec, 63(1), 63 - 8 Anti ICAM-1 (CD 54) monoclonal antibody reduces inflammatory changes in experimental bacterial meningitis; Weber JR et al.; We investigated whether monoclonal antibodies directed against intracellular adhesion molecule 1 (ICAM-1 mAb) inhibit brain edema, increase of intracranial pressure (ICP), regional cerebral blood flow (rCBF) and recruitment of white blood cells (WBC) into the cerebrospinal fluid (CSF) in the rat model of the early phase of bacterial meningitis . Brain edema was assessed by brain water content determinations . rCBF measured by laser Doppler flowmetry and ICP were recorded continuously for 6 h after intracisternal challenge . Meningitis was induced with pneumococcal cell walls (PCW) . Increase of ICP and brain water content were significantly inhibited (P <0.05) by intravenous treatment with ICAM-1 mAb (TM-8, 1 mg/kg) . Furthermore, ICAM-1 mAb treatment profoundly attenuated (P <0.05) rCBF increase and WBC invasion into the CSF . These results suggest that the ICAM-1 pathway is critically involved in the early phase of bacterial meningitis. Br J Biomed Sci, 1995 Dec, 52(4), 321 - 2 Bacterial capsules: a simple method for demonstration under the light microscope; Okeke IN et al.; It is sometimes desirable to demonstrate bacterial capsules during the routine examination of clinical isolates . Apart from the Indian ink method, methods of demonstrating bacterial capsules are not only tedious but are often non-reproducible . A combined positive-negative capsule staining procedure which is simple, rapid and reproducible is described. Plant Mol Biol, 1995 Dec, 29(5), 959 - 68 Characterization of tobacco plants expressing a bacterial salicylate hydroxylase gene; Friedrich L et al.; Transgenic tobacco plants that express the bacterial nahG gene encoding salicylate hydroxylase have been shown to accumulate very little salicylic acid and to be defective in their ability to induce systemic acquired resistance (SAR) . In recent experiments using transgenic NahG tobacco and Arabidopsis plants, we have also demonstrated that salicylic acid plays a central role in both disease susceptibility and genetic resistance . In this paper, we further characterize tobacco plants that express the salicylate hydroxylase enzyme . We show that tobacco mosaic virus (TMV) inoculation of NahG tobacco leaves induces the accumulation of the nahG mRNA in the pathogen infected leaves, presumably due to enhanced stabilization of the bacterial mRNA . SAR-associated genes are expressed in the TMV-infected leaves, but this is localized to the area surrounding necrotic lesions . Localized acquired resistance (LAR) is not induced in the TMV-inoculated NahG plants suggesting that LAR, like SAR, is dependent on SA accumulation . When SA is applied to nahG-expressing leave's SAR gene expression does not result . We have confirmed earlier reports that the salicylate hydroxylase enzyme has a narrow substrate specificity and we find that catechol, the breakdown product of salicylic acid, neither induces acquired resistance nor prevents the SA-dependent induction of the SAR genes. Postgrad Med J, 1995 Dec, 71(842), 745 - 6 Bacterial meningitis after MMR immunisation; Riordan FA et al.; Two children developed bacterial meningitis within five days of measles-mumps-rubella (MMR) immunisation . Diagnosis was delayed because symptoms were attributed to the vaccine, although both had a raised C-reactive protein . Fever or rash within five days of MMR vaccination are unlikely to be due to the vaccine and a raised C-reactive protein suggests bacterial infection. Br J Surg, 1995 Dec, 82(12), 1663 - 7 Preoperative total parenteral nutrition is not associated with mucosal atrophy or bacterial translocation in humans; Sedman PC et al.; Concerns have recently been expressed at suggestions that postoperative sepsis may be more common in patients who have received preoperative total parenteral nutrition (TPN) . The mechanism suggested for this is that TPN causes intestinal mucosal atrophy leading to increased bacterial translocation from the gut as a source of systemic sepsis . This hypothesis was examined in 203 patients who had an elective laparotomy, 28 of whom required at least 10 days of preoperative TPN . Neither mucosal atrophy nor bacterial translocation was more common in parenterally fed patients than in enterally fed controls . In humans theoretical concerns about the adverse effects of TPN on intestinal integrity are unfounded. J Neurochem, 1995 Dec, 65(6), 2690 - 8 Induction of prostanoid biosynthesis by bacterial lipopolysaccharide and isoproterenol in rat microglial cultures; Minghetti L et al.; We have used purified microglial cultures obtained from neonatal rat cerebral cortex to investigate the ability of microglia to release prostanoids after exposure to bacterial lipopolysaccharide, a classic macrophage activator . Release of prostaglandin E2, prostaglandin D2, and thromboxane A2 was low in basal conditions and increased in a dose- and time-dependent way upon lipopolysaccharide treatment (1-100 ng/ml), by a mechanism requiring de novo protein synthesis . When compared with astrocytes, microglial cells appeared to respond more effectively to lipopolysaccharide, being able to release prostanoids after exposure to a 100-fold lower concentration of lipopolysaccharide . In addition to prostanoids, we also measured the release of leukotriene B4; although lipopolysaccharide failed to stimulate leukotriene B4 release by microglial cells, it doubled the basal production in astrocytes . Lipopolysaccharide enhanced the release of preloaded {3H}arachidonic acid from microglial membrane phospholipids by a mechanism inhibited by the protein synthesis inhibitor cycloheximide, which suggests that the increased availability of arachidonic acid contributed to the enhanced prostanoid production . Lipopolysaccharide, however, also stimulated prostanoid synthesis by inducing cyclooxygenase activity, as shown by determining the activity of newly synthesized enzyme after inactivating the endogenous enzyme with aspirin and by assessing the level of the inducible form of cyclooxygenase by western blot analysis . Among the mechanisms potentially involved in the regulation of microglial prostanoid production, we studied the effect of beta-adrenergic receptor activation . The beta-agonist isoproterenol was inactive by itself but doubled the effect of lipopolysaccharide.(ABSTRACT TRUNCATED AT 250 WORDS) Int Arch Allergy Immunol, 1995 Dec, 108(4), 327 - 33 Genetic detoxification of bacterial toxins: a new approach to vaccine development; Rappuoli R et al.; Chemically detoxified bacterial toxins (toxoids) have been successfully used as vaccines for the prevention of many bacterial infectious diseases . Today, nontoxic derivatives of bacterial toxins can be obtained by mutagenesis of the toxin genes . These genetically inactivated toxins are superior to the classical toxoids both in safety and in immunogenicity and therefore they should replace the old toxoids in the existing vaccines . In addition, they represent a novel class of immunogens with unique properties, some of which may be used for innovative approaches to vaccination. J Am Dent Assoc, 1995 Dec, 126(12), 1634 - 9 Reducing bacterial aerosol contamination with a chlorhexidine gluconate pre-rinse; Logothetis DD et al.; The authors compared the effects of chlorhexidine gluconate, an antiseptic mouthwash with essential oils and water on the bacterial aerosol contamination generated by an air polishing device . Patients rinsed with one of the three solutions before treatment . Bacterial counts collected during the treatment indicate that the chlorhexidine pretreatment rinse was significantly more effective than the other solutions in reducing bacterial aerosols. Cancer Res, 1995 Dec 1, 55(23 Suppl), 5968s - 5972s Bacterial expression of a kemptide fusion protein facilitates 32P labeling of a humanized, anti-carcinoembryonic antigen (hMN-14) antibody fragment; Leung SO et al.; Despite the potential advantages of 32P over other isotopes for radioimmunotherapy, its development as a therapeutic has been hindered by the difficulty of the labeling chemistry . Recently, a heptapeptide {Kemptide (KPT)} has been chemically conjugated to antibodies, and the conjugates have successfully been labeled with 32P enzymatically by using bovine protein kinase . By using genetic engineering, we have produced a chimera (Fab.KPT) consisting of the Fab' moiety of the complementarity-determining region-grafted anti-carcinoembryonic antigen-monoclonal antibody, MN14, and a heptapeptide derivative of KPT (Trp-Arg-Arg-Ala-Ser-Leu-Gly) . The recombinant protein was expressed in Escherichia coli as a soluble secretory product . The presence of the KPT derivative downstream of the COOH terminus of the hinge region did not impair the binding affinity of the antibody fragment . The Fab.KPT was enzymatically phosphorylated with 32P by bovine protein kinase, without significant effect on the resultant immunoreactivity; 100% of the 32P-labeled Fab.KPT was complexed with liquid carcinoembryonic antigen . The 32P-labeled humanized MN-14 Fab.KPT is expected to have longer blood circulation half-life, allowing for an improved therapeutic efficacy in radioimmunotherapy. Blood, 1995 Dec 1, 86(11), 4184 - 93 Critical involvement of transmembrane tumor necrosis factor-alpha in endothelial programmed cell death mediated by ionizing radiation and bacterial endotoxin; Eissner G et al.; In this report, we show that ionizing radiation (IR) at a clinically relevant dose (4 Gy) causes apoptosis in macrovascular and microvascular human endothelial cells . Treatment of irradiated cells with a low dose of bacterial endotoxin (LPS), similar to the levels observed in serum during endotoxemia, enhanced the rate of apoptosis, although LPS alone was unable to induce programmed cell death . The cytokine and endotoxin antagonist interleukin-10 (IL-10) reduced the rate of LPS + IR-induced apoptosis to levels obtained with irradiation alone . Using neutralizing antibodies against tumor necrosis factor-alpha (TNF), we could show crucial involvement of TNF in the LPS-mediated enhancement of IR-induced apoptosis, but not in the IR-induced apoptosis per se . However, further analysis strongly suggested the transmembrane form of TNF (mTNF), but not soluble TNF, to be accountable for the LPS-mediated cytotoxic effects . Studies with anatagonistic receptor specific antibodies clearly showed that TNF receptor type I (TR60) is essential and sufficient to elicit this effect . These findings are of potential clinical importance because they may disclose a relevant mechanism that leads to endothelial damage after radiotherapy or total body irradiation used for conditioning in bone marrow transplantation and that may thus contribute to transplant related complications, especially in association with endotoxemia or related inflammatory states. EMBO J, 1995 Nov 15, 14(22), 5679 - 89 A yeast transcription factor bypassing the requirement for SBF and DSC1/MBF in budding yeast has homology to bacterial signal transduction proteins; Morgan BA et al.; The transcription factors SBF and DSC1/MBF bind SCB and MCB promoter elements, respectively, and are essential for the cell cycle progression of Saccharomyces cerevisiae through the control of G1 cyclin gene expression . We isolated a gene (BRY1; Bacterial Response regulator in Yeast) able to activate either MCB or SCB promoter elements on a reporter plasmid which, when overexpressed, can bypass the normally essential requirement for SBF and DSC1/MBF by the stimulation of CLN1 and CLN2 expression . In the case of CLN2 at least, this expression depends upon the MCB and SCB promoter elements . In wild-type yeast, the disruption of BRY1 has no apparent phenotype, but under conditions where the activities of SBF and DSC1/MBF are reduced, BRY1 becomes essential . Our data imply the existence of a third pathway affecting cyclin expression . BRY1 is the same gene as SKN7 which has significant sequence homology to the receiver domains found in response regulator proteins from the bacterial two-component signal transduction pathways . SKN7 is thought to affect cell wall structure, and when highly overexpressed we find that BRY1/SKN7 is lethal perhaps because of perturbations in cell wall biosynthesis . The lethality is partially rescued by genes from the protein kinase C pathway, but genetic data imply that BRY1/SKN7 and protein kinase C are not in the same pathway . Our results suggest that Bry1/Skn7 can influence the expression of MCB- and SCB-driven gene expression in budding yeast, perhaps including genes involved in cell wall metabolism, via a two-component signal transduction pathway which activates Bry1/Skn7 in response to an unidentified signal. Arch Biochem Biophys, 1995 Nov 10, 323(2), 438 - 46 Comparison of isoniazid oxidation catalyzed by bacterial catalase-peroxidases and horseradish peroxidase; Hillar A et al.; The physical properties and activities of the purified catalase-peroxidase hydroperoxidase I (HPI) of Escherichia coli (EcHPI) and HPI with a carboxyl-terminal extension of Mycobacterium tuberculosis (MtHPI-e) are compared to those of commercial preparations of horseradish peroxidase (HRP) . The catalase-peroxidase proteins had similar absorption spectra and differed primarily in that MtHPI-e has a higher peroxidatic to catalatic activity ratio than EcHPI . Trypsin cleavage of MtHPI-e resulted in the formation of an active catalase-peroxidase lacking the carboxyl-terminal extension . The three enzymes, HRP, MtHPI-e, and EcHPI, mediated the isoniazid- and H2O2-dependent production of radical species, as detected by nitroblue tetrazolium reduction . A constant flux of H2O2, generated in situ from glucose oxidase and glucose was used . MtHPI-e was more effective at isoniazid-dependent radical production than EcHPI and HRP . Similar qualitative results were obtained by staining nondenaturing polyacrylamide gels for activity with nitroblue tetrazolium in the presence of isoniazid and H2O2 . The absorbance spectrum of HRP exhibited changes during incubation with isoniazid and H2O2 consistent with the formation of several typical reaction intermediates, whereas the catalase-peroxidases exhibited no distinct spectral changes . The results suggest that the sensitivity of M . tuberculosis to isoniazid may be the result of isoniazid-dependent radical formation by the catalase-peroxidase in the absence of other catalase activities to remove substrate H2O2. Antonie Van Leeuwenhoek, 1995 Nov, 68(4), 317 - 27 Stimulation of conversion rates and bacterial activity in a silage-fed two-phase biogas process by initiating liquid recirculation; Jarvis A et al.; The effects of liquid recirculation on a liquefaction-acidogenic reactor in an anaerobic two-phase digesting system operating with grass-clover silage was studied during 40 days after initiating recirculation of effluent from the methanogenic reactor to the liquefaction-acidogenic reactor . An increase in alkalinity and, thus, an increase in pH from 5.2 to 6.0 occurred in the liquefaction-acidogenic reactor . During the same period, a 10-fold increase (from 0.2 to 1.9 g.l-1.h-1) in the degradation rate of mannitol and an almost 9-fold increase in the activity of hydrogenotrophic methanogens was observed . The estimated number of these bacteria increased by one order of magnitude . The average degradation rate of lactate increased 3-fold, probably as a consequence of the more efficient hydrogen consumption by the hydrogenotrophic methanogens . An observed increase in net mineralization of organic nitrogen compounds was probably the main reason for an enhanced net production of organic acids (from 0.2 to 0.9 g.l-1.d-1) . The liquefaction of cellulose and hemicellulose was low from the start of recirculation (3% and 20% reduction, respectively) and did not seem to be affected by the liquid recirculation . This was in accordance with the low number of cellulose degraders (4.0 x 10(2) counts.ml-1) observed . The results from this investigation show that the initiation of liquid recirculation in silage-fed two-phase biogas processes will stimulate the activity of hydrogenotrophic methanogens in the liquefaction-acidogenic reactor . This will lead to more thermodynamically favourable conditions for acidification reactions which are dependent upon interspecies transfer of reducing equivalents. Dev Comp Immunol, 1995 Nov-Dec, 19(6), 473 - 82 Macrophage activating factor(s) secreted by mitogen stimulated goldfish kidney leukocytes synergize with bacterial lipopolysaccharide to induce nitric oxide production in teleost macrophages; Neumann NF et al.; Recent studies in our laboratory demonstrated that fish macrophages produce nitric oxide . To elucidate the mechanisms which regulate nitric oxide production in teleosts, we examined whether macrophage activating factors (MAFs) secreted by mitogen stimulated leukocytes, induced nitric oxide production in a long-term cultured macrophage cell line and in primary cultures of kidney macrophages from the goldfish . The results indicate that both primary and long term cultured goldfish macrophages produce nitric oxide in response to MAF or bacterial lipopolysaccharide (LPS), and co-stimulation with both factors results in a synergistic induction of nitric oxide production . MAF that induced nitric oxide production were present in leukocyte supernatants as early as 24 h after addition of mitogens to cell cultures . The production of MAF was dependent upon the incubation temperature, presence of serum in the culture medium and duration of incubation: maximal MAF activity was detected in 72-96 h supernatants raised in media with serum at 30 degrees C . MAF-induced nitric oxide production by long term cultured macrophages was inhibited by 1000 microM NG-monomethyl-L-arginine or amino-guanidine, indicating an L-arginine-dependent metabolic pathway for the production of the reactive nitrogen intermediates in teleosts . The biochemical events of cytokine induced nitric oxide production by teleost macrophages appear to be similar to those of mammalian macrophages. Vopr Virusol, 1995 Nov-Dec, 40(6), 282 - 4 {Detection of viral-bacterial factors responsible for infertility in couples}; L'vov ND et al.; A high prevalence of genital infections was revealed in patients suffering for a long time from sterility . The inflammatory process was found to predominate in tubal sterility . In other forms of sterility with asymptomatic urogenital infections the couples are frequently unaware of the disease and are not properly examined . Genital inflammations not diagnosed for many years augment the endocrine disorders and stimulate the development of autoimmune states . Today, a mixed viral/bacterial urogenital infection is the principal cause of reproductive disorders. Kekkaku, 1995 Nov, 70(11), 639 - 44 {The role of some cellular components of bacterial parasites in determining the incidence of tuberculosis: studies on mycobacterial antigens, with special reference to mycobacterial immunoreactive ribosomal and secreted proteins}; Yamada T et al.; Tuberculosis remains as major disease, affecting more than 20 million people . The elimination of the disease with vaccination, rapid diagnosis, and and efficient therapy is an important objective of our study . To realize the objective, the characterization of antigens is essential . We have chosen two kinds of antigens for our study, the ribosomal antigens and and an antigenic proteins secreted by mycobacteria . The biochemical and immunological characterization of ribosomal fraction was carried out . Ribosomal proteins were purified and assessed for DTH reaction . The N-terminal amino acids sequences were determined . Total structures of S19, S7 and S12 in 30S and L7/L12 in 50S subunits were elucidated . L7/L12 had 66% homology with analogue from S . griseus which showed GTPase activity in protein synthesis . This protein was secreted in culture medium and induced strong DTH . Secreted antigenic proteins are of great interest for us . Secreted antigens may be recognized rapidly by immune system and therefore may induce rapid and high level immune response . It is also expected that it may contain protective antigens, since live BCG protect disease more efficiently than heat killed BCG . We have determined and published the total structure of four proteins (MPB64, MPB70, MPB57 and alpha antigen) . We attempted to utilize this antigen for the diagnosis and the design of vaccine . The structures of alpha antigens from M . avium, M . intracellulare, M . scrofulaceum, M . kansasii and BCG were determined and its potential for application to diagnosis was presented . Using the operon of M . kansasii, alpha antigen and V3 region of HIV-1 were expressed by recombinant BCG which induced CTL in mice. Vet Immunol Immunopathol, 1995 Nov, 49(1-2), 51 - 60 Interleukin-6 secretion by bacterial lipopolysaccharide-stimulated bovine alveolar macrophages in vitro; Jian ZJ et al.; Interleukin-6 (IL-6) is a pluripotent cytokine that may play a role in pulmonary defense against bacterial pathogens . We have quantitated the response of bovine alveolar macrophages (bAM) to bacterial lipopolysaccharide (LPS; E . coli 055: B5) in vitro using the IL-6 sensitive 7TD1 cell line . Bacteria LPS in the absence of serum induced IL-6 secretion from bAM (1 x 10(6) ml-1) over a range of LPS concentrations from 10 ng ml-1 to 10 micrograms ml-1 . This resulted in IL-6 levels ranging from approximately 5 to over 200 U ml-1.IL-6 secretion by from approximately 5 to over 200 U ml-1.IL-6 secretion by LPS-stimulated bAM was increased by 24 h poststimulation, and continued to increase up to 72 h after stimulation . Fetal bovine serum (FBS, 1% vol/vol; 320 micrograms ml-1) enhanced IL-6 secretion from macrophages in the presence of LPS by approximately 10-fold compared with LPS alone . A bovine serum fraction (1 microgram ml-1 protein) prepared using ion-exchange chromatography also markedly enhanced IL-6 secretion versus LPS alone . The stimulatory effect of IL-6-like activity in the bAM supernatants was neutralized by an anti-human IL-6 polyclonal antibody . Northern blot analysis revealed increased IL-6 mRNA at 2 h poststimulation with LPS + FBS, peak levels at 4 h, and levels were decreased by 6 h poststimulation . Results suggest that IL-6 is secreted by bovine alveolar macrophages, and that bacterial LPS and serum components synergize to produce this response. J Clin Immunol, 1995 Nov, 15(6), 338 - 48 CD4+ Th2 cell response cytokine production in bacterial meningitis; Raziuddin S et al.; There has been a growing body of evidence suggesting that CD4+ Th1/Th2 cell responses participate in pathologic and immunologic processes in infectious disease . Bacterial meningitis is a fatal disease of children and is associated with a spectrum of clinical syndromes . This study provides evidence of CD4+ enhanced interleukin (IL)-4 and IL-6 but decreased IL-2 and interferon-gamma (IFN-gamma) production, the induction of characteristic Th2 cell response cytokines in bacterial meningitis, which may play an important role in disease mechanism . Additionally, monocyte-induced enhanced IL-6, IL-8, and tumor necrosis factor-alpha production may be associated with distinct clinical features such as fever, seizures, and neurological sequelae . A striking finding was also the highly deficient monocyte-induced granulocyte-macrophage colony-stimulating factor production . Of particular interest, the CD(8+)-enhanced IFN-gamma production may be required for the cytolytic activity or protective response to be maintained in this disease . Taken together, these data reveal that monocytes and CD4+ (Th2) and CD8+ subsets produce distinct cytokines in bacterial meningitis, which may exert an immunoregulatory and immunopathologic effect and thus mediate some of the clinical manifestations of the disease. Trends Microbiol, 1995 Nov, 3(11), 441 - 5 In vitro models of the blood-brain barrier to study bacterial meningitis; Townsend GC et al.; In vitro models of the blood-brain barrier involving culturing cerebral microvascular endothelial cells may provide information critical to understanding diseases of the central nervous system, such as bacterial meningitis, that would be difficult to obtain from clinical studies or from in vivo models . These models may also identify targets for therapeutic intervention. J Invertebr Pathol, 1995 Nov, 66(3), 287 - 92 In vivo chemoactivation of oyster hemocytes induced by bacterial secretion products; Alvarez MR et al.; Movements of tissue hemocytes in the Eastern oyster Crassostrea virginica were monitored and quantified by image analysis of sections following inoculation with agar cores containing Escherichia coli or cell-free medium on which the bacteria had previously grown . Hemocytes respond to the presence of live bacteria by accumulating in widely dispersed areas of tissue surrounding the gut and digestive diverticula . The response is rapid and evident within 40 min, is maximal at 1 hr, and declines by 3 hr after inoculation . Sterile implanted agar cores do not produce a response . Bacteria killed with ozone elicit a response when inoculated together with the medium on which they had grown while bacteria killed by heat or formalin do not . Killed bacteria suspended in saline fail to stimulate hemocyte chemokinesis . Cell-free medium applied externally produces a response equal to that measured with live bacteria inoculated internally . Extraction of bacteria-free medium with hexane does not significantly reduce hemocyte chemokinesis . Digestion of bacteria-free medium with pronase completely eliminates chemokinesis . Molecular filtrates of bacteria-free medium induce maximal chemokinetic response at molecular weight as low as 1 kDa . These data show that the oyster hemocyte activators produced by E . coli are most likely low-molecular-weight polypeptides which diffuse from the site of inoculation and can pass through the intact external surface epithelium to induce a chemokinetic response. Burns, 1995 Nov, 21(7), 521 - 5 Influence of a changed care environment on bacterial colonization of burn wounds; Adeniran A et al.; This study investigated the influence of a conditioned care environment per se on bacterial colonization of burn wounds . Two cohorts of burn patients were treated in the successive years 1992 and 1993, the first group in a (permanent) purpose-designed unit and the second in wards of traditional 'open' design, during renovation of the unit . Patients who were admitted to the permanent and temporary units numbered 224 and 231 respectively, the groups being similar in features that generally influence the course and outcome of burn injuries . The principles and practice of treatment by the burn care team remained the same in both years . No significant difference in wound colonization rates was found between the two groups . We conclude that while the other known advantages of managing burn patients in purpose-designed units remain valid, a conditioned care environment per se does not influence bacterial colonization rates of burn wounds. Microbiology, 1995 Nov, 141 ( Pt 11), 2839 - 48 Chimeric FimH adhesin of type 1 fimbriae: a bacterial surface display system for heterologous sequences; Pallesen L et al.; The FimH adhesin of type 1 fimbriae has been tested as a display system for heterologous protein segments on the surface of Escherichia coli . This was carried out by introduction of restriction site handles (BglII sites) in two different positions in the fimH gene, followed by in-frame insertion of heterologous DNA segments encoding two reporter sequences . In the selected positions such insertions did not significantly alter the function of the FimH protein with regard to surface location and adhesive ability . The system seemed to be quite flexible, since chimeric versions of the FimH adhesin containing as many as 56 foreign amino acids were transported to the bacterial surface as components of the fimbrial organelles . Furthermore, the foreign protein segments were recognized by insert-specific antibodies when expressed within chimeric proteins on the surface of the bacteria . The results from this feasibility study point to the possibility of using the FimH adhesin as a general surface display system for sizeable protein segments. Eur J Biochem, 1995 Nov 1, 233(3), 766 - 71 The functional integration of a polytopic membrane protein of Escherichia coli is dependent on the bacterial signal-recognition particle; Macfarlane J et al.; In eukaryotes, the cotranslational targeting of proteins to the endoplasmic reticular membrane is initially mediated by the signal-recognition particle (SRP), a ribonucleoprotein complex consisting of the 7SL RNA and six protein subunits . Since the discovery of sequence homology between (a) the Escherichia coli 4.5S RNA (Ffs) and 7SL RNA, and (b) the E . coli P48 (Ffh) and SRP 54-kDa subunit, more evidence has been obtained that E . coli also possesses an SRP-type pathway that acts in the translocation of secreted proteins . Such a pathway could possibly be involved in the cotranslational integration of hydrophobic membrane proteins that cannot be effectively targeted post-translationally due to folding and aggregation . In this study, we report that disruption of the E . coli SRP complex with a dominant lethal 4.5S RNA mutant in vivo prevents functional membrane integration of the E . coli lactose permease (LacY) . Likewise, depletion of the P48 (Ffh) protein also results in a decrease in the amount of functional LacY inserted into the E . coli plasma membrane . In direct contrast, inhibition of SecA function does not affect LacY integration . These results suggest a major function of the bacterial SRP in the targeting and subsequent integration of hydrophobic membrane proteins as opposed to SecA mediating the post-translational targeting of secretory proteins. J Fam Pract, 1995 Nov, 41(5), 443 - 9 Treatment of bacterial vaginosis: a comparison of oral metronidazole, metronidazole vaginal gel, and clindamycin vaginal cream; Ferris DG et al.; BACKGROUND . Treatment options for bacterial vaginosis are numerous . The purpose of this study was to compare the efficacy of oral metronidazole, metronidazole vaginal gel, and clindamycin vaginal cream for the treatment of bacterial vaginosis using traditional clinical and laboratory methods, as well as a new DNA probe test . We also determined the percentage of patients receiving each treatment who developed posttreatment vaginal candidiasis, a potential complication of treating bacterial vaginosis . METHODS . One hundred one women in whom bacterial vaginosis was diagnosed by standard criteria were randomly assigned to receive: oral metronidazole 500 mg twice daily for 1 week, 0.75% metronidazole vaginal gel 5 g twice daily for 5 days, or 2% clindamycin vaginal cream 5 g once daily for 7 days . Women with coexisting vulvovaginal candidiasis or vaginal trichomoniasis were excluded . Tests of cure by vaginal saline wet prep and potassium hydroxide microscopic examinations, Gram's stain, pH and DNA probe tests for Gardnerella vaginalis and Candida species were scheduled 7 to 14 days following treatment . RESULTS . There were no statistically significant differences in cure rates for oral metronidazole (84.2%), metronidazole vaginal gel (75.0%), or clindamycin vaginal cream (86.2%) (chi 2 = 1.204, df = 2, P = .548) using traditional clinical and laboratory criteria . Cure rates were lower based on DNA testing, indicating that Gardnerella vaginalis may remain after a clinical cure . This would explain cases of recurrent disease . Posttreatment vulvovaginal candidiasis was experienced by 12.5% of subjects treated with oral metronidazole, 14.8% of subjects treated with clindamycin vaginal cream, and 30.4% of subjects treated with metronidazole vaginal gel (chi 2 = 2.607, df = 2, P = .272) . CONCLUSIONS . Oral metronidazole, metronidazole vaginal gel, and clindamycin vaginal cream achieved nearly equivalent cure rates for the treatment of bacterial vaginosis . Patients treated with these agents experienced similar rates of posttreatment vulvovaginal candidiasis, but those using the intravaginal products reported being more satisfied with the treatment. Crit Care Med, 1995 Nov, 23(11), 1829 - 34 Elastin fibers and the diagnosis of bacterial pneumonia in the adult respiratory distress syndrome; Shepherd KE et al.; OBJECTIVE: It has been inferred from previous work that 40% potassium hydroxide preparations of lower respiratory tract secretions that demonstrate elastin fibers have a 100% specificity and positive predictive value in diagnosing bacterial pneumonia in intubated, mechanically ventilated patients without the adult respiratory distress syndrome (ARDS) . Our aim was to assess the specificity of 40% potassium hydroxide preparations in diagnosing bacterial pneumonia in patients with ARDS and suspected pneumonia . DESIGN: Prospective, case-referral clinical study . SETTING: Referral hospital . PATIENTS: Of 24 patients with ARDS who were intubated and mechanically ventilated with suspected bacterial pneumonia, 22 were assessable and evaluated for this report . INTERVENTIONS: Tracheo-bronchial aspirates were obtained from all patients and analyzed for elastin fibers using 40% potassium hydroxide . MEASUREMENTS AND MAIN RESULTS: Of the 22 assessable patients, ten patients did not have a complicating bacterial pneumonia . Six of these ten patients had potassium hydroxide preparations that demonstrated elastin fibers (false positives) . The other four patients had preparations that did not demonstrate elastin fibers (true negatives) . Specificity was 40% . CONCLUSION: Elastin fiber preparations are not specific for diagnosing bacterial pneumonia in patients with ARDS. Am J Respir Crit Care Med, 1995 Nov, 152(5 Pt 1), 1549 - 54 GRO alpha and interleukin-8 in Pneumocystis carinii or bacterial pneumonia and adult respiratory distress syndrome; Villard J et al.; Polymorphonuclear leukocytes (PMN) are the predominant inflammatory cells recruited in acute lung injury . This study compares the concentration of interleukin-8 (IL-8) to those of GRO alpha, both of which are CXC chemokines, in bronchoalveolar lavage fluid (BALF) in three acute pathologic states: bacterial pneumonia (BPN); adult respiratory distress syndrome (ARDS); and Pneumocystis carinii pneumonia (PCP) . Levels of both IL-8 and GRO alpha were below 5 pg/ml in 16 nonsmoking volunteers who served as controls . Despite more than twice as many neutrophils in the BALF of the BPN group (n = 12) than in the group with ARDS (n = 13), both groups had similar levels of IL-8, of 569 +/- 120 pg/ml and 507 +/- 96 pg/ml, respectively . The GRO alpha concentrations in the BPN and ARDS patients were respectively 3.3 and 3.4 times those of IL-8, reaching 1,870 +/- 314 pg/ml for the BPN and 1,699 +/- 377 for the ARDS patients . In the PCP group (n = 48, 45 human immunodeficiency virus {HIV}-positive, 3 HIV-negative), GRO alpha levels (897 +/- 172 pg/ml) were sevenfold higher than IL-8 levels (123 +/- 40 pg/ml) . In all pathologic states there was a good correlation between GRO alpha and IL-8 (r = 0.53, p = 0.0001) . GRO alpha or IL-8 both correlate with the absolute neutrophil number/ml when all groups were studied together (r = 0.52, p = 0.0001) . Only in the PCP and ARDS groups did IL-8 correlate with the PMN number.(ABSTRACT TRUNCATED AT 250 WORDS) Exp Parasitol, 1995 Nov, 81(3), 284 - 91 Wolbachia pipientis: bacterial density and unidirectional cytoplasmic incompatibility between infected populations of Aedes albopictus; Sinkins SP et al.; Unidirectional cytoplasmic incompatibility is seen when certain Wolbachia-infected insect populations are crossed . Two hypotheses might explain this phenomenon: superinfections with mutually incompatible strains of Wolbachia producing incompatibility when crossed to individuals infected with only a single bacterial strain or, alternatively, a bacterial dosage model, with differences in Wolbachia densities responsible for the incompatibility . A quantitative PCR assay was set up as a general method to compare Wolbachia densities between populations . Using this assay in unidirectionally incompatible stocks of the mosquito Aedes albopictus, we have determined that densities are significantly higher in Houston than in the Mauritius and Koh Samui stocks . This is consistent with a dosage model for the observed crossing patterns, but does not rule out the possibility that superinfection is the primary cause of the incompatibility. J Surg Res, 1995 Nov, 59(5), 596 - 600 Fibronectin on the surface of biliary drain materials--a role in bacterial adherence; Yu JL et al.; The present study deals with the demonstration of deposited fibronectin (Fn) on the surfaces of implanted biliary drain materials and the role of deposited Fn in promotion of bacterial adherence . Rubber pieces that had been implanted in the biliary tracts of rats for 4 weeks were retrieved and the following approaches employed for further investigations: (1) adherence of {methyl-3H}thymidine-labeled Escherichia coli to implanted and unimplanted rubber pieces; (2) blocking the adherence of radiolabeled bacteria with anti-Fn antibodies; (3) detection of deposited Fn by 125I-labeled anti-Fn IgG; and (4) immunoblotting of the surface eluate from implanted rubber pieces . The results show that in the presence of serum, plasma, or bile, the number of E . coli cells adherent to implanted rubber pieces was 10 times higher than that adherent to the unimplanted pieces (P < 0.001) and that the adherence was reduced by pretreatment of implanted pieces with anti-Fn antibodies . Furthermore, the implanted pieces appeared to have a high affinity for 125I-labeled rabbit anti-Fn IgG rather than the 125I-IgG without anti-Fn fraction . Fn was also found in the surface eluate of implanted pieces by immunoblotting of the eluate . The results in the present study suggest that Fn may be involved in implant-associated infections in the biliary tract. FEBS Lett, 1995 Oct 30, 374(2), 257 - 61 The GA module, a mobile albumin-binding bacterial domain, adopts a three-helix-bundle structure; Johansson MU et al.; We present the first study of the secondary structure and global fold of an albumin-binding domain . Our data show that the GA module from protein PAB, an albumin-binding protein from the anaerobic bacterial species Peptostreptococcus magnus, is composed of a left-handed three-helix bundle . The helical regions were identified by sequential and medium range NOEs, values of NH-C alpha H coupling constants, chemical shift indices, and the presence of slowly exchanging amide protons, as determined by NMR spectroscopy . In addition, circular dichroism studies show that the module is remarkably stable with respect to both pH and temperature. J Biol Chem, 1995 Oct 27, 270(43), 25328 - 31 Increase of solubility of foreign proteins in Escherichia coli by coproduction of the bacterial thioredoxin; Yasukawa T et al.; Eukaryotic proteins are frequently produced in Escherichia coli as insoluble aggregates . This is one of the barriers to studies of macromolecular structure . We have examined the effect of coproduction of the E . coli thioredoxin (Trx) or E . coli chaperones GroESL on the solubility of various foreign proteins . The solubilities of all eight vertebrate proteins examined including transcription factors and kinases were increased dramatically by coproduction of Trx . Overproduction of E . coli chaperones GroESL increased the solubilities of four out of eight proteins examined . Although the tyrosine kinase Lck that was produced as an insoluble form and solubilized by urea treatment had a very low autophosphorylating activity, Lck produced in soluble form by coproduction of Trx had an efficient activity . These results suggest that the proteins produced in soluble form by coproduction of Trx have the native protein conformation . The mechanism by which coproduction of Trx increases the solubility of the foreign proteins is discussed. Gene, 1995 Oct 16, 164(1), 153 - 6 Aspartyl-tRNA synthetase of the hyperthermophilic archaeon Pyrococcus sp . KOD1 has a chimerical structure of eukaryotic and bacterial enzymes; Imanaka T et al.; The aspartyl-tRNA synthetase (AspRS)-encoding gene from the archaeon, Pyrococcus sp . KOD1 (KOD1), was cloned and sequenced, and expressed in Escherichia coli . The purified AspRS possessed an aminoacyation activity for tRNA extracted from KOD1 . Analysis of the deduced amino-acid sequence (438 aa, 50,893 Da) revealed that the AspRS of KOD1 is a chimerical protein of bacteria and eukarya . Regional analysis showed high sequence similarity to higher eukaryotic enzymes in the central and C-terminal regions which are important for catalytic activity of the enzyme . In contrast, the N-terminal portion exhibits bacterial features and does not possess the higher eukaryotic sequence which is involved in high molecular weight (HMW) complex formation . These results suggest that archaeon AspRS has a eukaryotic-type catalytic mechanism without forming the HMW complex . This is the first example which shows that an archaeal protein possesses eukaryotic and bacterial features. Eur J Biochem, 1995 Oct 15, 233(2), 506 - 13 Bacterial expression of the catalytic domain of 3-hydroxy-3-methylglutaryl-CoA reductase (isoform HMGR1) from Arabidopsis thaliana, and its inactivation by phosphorylation at Ser577 by Brassica oleracea 3-hydroxy-3-methylglutaryl-CoA reductase kinase; Dale S et al.; The catalytic domain of 3-hydroxy-3-methylglutaryl-CoA reductase isoform 1 (HMGR1cd) from Arabidopsis thaliana has been expressed in Escherichia coli in a catalytically active form and purified . The high efficiency of the bacterial expression system together with the simplicity of the purification procedure used in this study resulted in the attainment of large quantities of pure enzyme (about 5 mg/l culture) with a final specific activity of up to 17 U/mg . This specific activity is higher than that reported to date for any 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) purified from a plant source . HMGR1cd activity was completely blocked by the HMGR inhibitor mevinolin (IC50 = 12.5 nM) . No significant differences were observed between the Km values of HMGR1cd for NADPH (71 +/- 7 microM) and (S)-3-hydroxy-3-methylglutaryl-CoA (8.3 +/- 1.5 microM) and those of pure HMGR preparations obtained from different plant sources . The purified HMGR1cd was reversibly inactivated by phosphorylation at a single site by Brassica oleracea HMGR kinase A, which is functionally related to the mammalian AMP-activated protein kinase . The site of phosphorylation is Ser577 in the complete sequence of A . thaliana HMGR1 . The results in this paper represent the first evidence that a higher plant HMGR is regulated by direct phosphorylation, at least in a cell-free system . Our results also reinforce the view that the AMP-activated protein kinase/SNF1 family is an ancient and highly conserved protein kinase system. J Biol Chem, 1995 Oct 13, 270(41), 24038 - 42 The N-terminal cytoplasmic tail of the aspartate receptor is not essential in signal transduction of bacterial chemotaxis; Chen X et al.; To determine the role in transmembrane signaling of the N-terminal peptide of the first transmembrane region of the aspartate receptor, it was subjected to extensive mutagenesis . Drastic changes did not alter the chemotactic ability of the receptor to aspartate significantly . Thus the cytoplasmic N terminus of the first transmembrane region does not play an essential role in transmembrane signaling, and the entire signal that is transmitted to the cytoplasmic domain must be sent through the second transmembrane region . This eliminates the models requiring an interaction of this N-terminal peptide with the remaining cytoplasmic portion of the receptor. Biochem Biophys Res Commun, 1995 Oct 13, 215(2), 517 - 23 cDNA cloning and bacterial expression of the human type I keratin 16; Paladini RD et al.; The human type I keratin 16 is constitutively expressed in a number of complex epithelial tissues, including skin, but is better known for its induction under conditions favoring enhanced proliferation or abnormal differentiation, including wound healing, psoriasis, and cancer . We cloned the coding sequence of human K16 by applying a coupled reverse transcription-polymerase chain reaction procedure to mRNAs prepared from cultured human skin keratinocytes . We then expressed the human K16 coding sequence in E . coli and purified the solubilized protein by anion-exchange chromatography . The recombinant protein recovered behaves similarly to human K14 (a related acidic keratin) on the anion-exchanger, co-migrates with native human K16 on SDS-PAGE (M(r) 48 kD), and reacts with antisera directed against human K16 . Based on the nucleotide sequence obtained and the properties of the corresponding recombinant protein, we conclude that we have cloned the coding portion of the human K16 cDNA . The sequence data obtained in this study is compared to earlier reports of the human K16 sequence, which are conflicting in many respects . The availability of K16 in a purified recombinant form will allow us to study how its properties may relate to its function during wound healing and in skin diseases. Biochemistry, 1995 Oct 10, 34(40), 13074 - 81 A bacterial luciferase reaction with a negative temperature coefficient attributable to protein-protein interaction; Sirokman G et al.; A yellow fluorescent protein (YFP) present in a strain of bioluminescent bacteria is shown here not only to modify the color and intensity of the emission, as already known and attributed to the interaction of YFP with a luciferase intermediate, but also remarkably to confer a negative temperature dependence to the in vitro system . The in vitro bioluminescence decay rate is actually independent of temperature in the range 5-25 degrees C, at approximately 1 microM YFP concentration . Several hypotheses are considered to explain this effect, based either on inactivation of YFP itself at higher temperatures or on its binding equilibrium with the luciferase intermediate . The first hypothesis is favored . Fluorescence anisotropy measurements show that YFP loses its chromophore at higher temperatures, but this alone cannot account for the negative temperature dependence . Gel chromatography shows the existence of an inactive YFP dimer, and the formation of more dimer at higher temperatures cannot be ruled out but is unlikely in our experimental conditions . Conformational changes may contribute to YFP inactivation . To our knowledge, there is no prior example of an enzymatic reaction in which the rate is slower at higher temperatures, within a physiological range. Indian J Lepr, 1995 Oct-Dec, 67(4), 363 - 74 Effect of T-cell depletion on bacterial multiplication and pattern of nerve damage in M . leprae-infected mice; Shetty VP et al.; Various mechanisms for nerve damage in tuberculoid leprosy have been proposed . A common feature amongst them is the crucial role played by T-cells . Therefore, the present study was designed to determine the role of T-cells in the induction of nerve damage in leprosy using two different protocols for obtaining graded levels of T-cell depletion: (i) Cyclosporine A, for depletion of T-helper cells and (ii) Anti Thy 1.2, for total depletion of T-cells . The findings indicate that the early changes seen in the unmyelinated fibres may not involve T-cells . However, the later stages of nerve damage associated with demyelination are dependent on T-cell responses. Eur Heart J, 1995 Oct, 16(10), 1448 - 50 Atypical presentation of adult Still's disease mimicking acute bacterial endocarditis; Zenagui D et al.; Adult Still's disease is a chronic, systemic disease of unknown origin . We describe the case of an otherwise healthy man with an uncommon presentation of Still's disease . A 38-year-old man presented with sore throat, fever, rash and arthritis . Laboratory findings showed that both erythrocyte sedimentation rate and ferritin had increased . Transoesophageal echocardiography revealed a vegetation involving the aortic leaflet . The diagnosis of Still's disease was made after the exclusion of infectious endocarditis, based upon the clinical picture, the high level of ferritin and the follow-up . The patient markedly improved after treatment with prednisone 1 mg . kg-1 . This controlled and then progressively reduced the disease; the drug was then withdrawn . This case illustrates that Still's disease can present with endocardial involvement mimicking acute bacterial endocarditis as a first clinical manifestation . The observation suggests that the presence of high ferritinaemia in a patient with some clinical criteria of Still's disease could lead to an early diagnosis. Zhonghua Nei Ke Za Zhi, 1995 Oct, 34(10), 680 - 2 {A randomized controlled clinical trial of sulperazone as compared with cefotaxime in the treatment of bacterial infections}; Li J et al.; A randomized controlled clinical trial of sulperazone as compared with cefotaxime was conducted . 207 patients with bacterial infections entered the study . The overall clinical efficacy rate of sulperazone and cefotaxime was 95.15% and 90.38% respectively . The bacterial clearance rate in the two groups was 84.7% and 80.6% respectively . The adverse drug reactions in both groups were mild with the rate of 7.77% and 8.65% respectively; there was no statistical difference between the two groups . The results of disc susceptibility test showed that the sensitive rate of the clinical isolates to sulperazone was 90.9%, being significantly higher than that of cefotaxime 69.3% (P < 0.001). J Ind Microbiol, 1995 Oct, 15(4), 339 - 46 Structure, function and immunochemistry of bacterial exopolysaccharides; Weiner R et al.; There has been much written on bacterial exopolysaccharides (EPS) and their role in virulence . Less has been published regarding EPS in free living species . This review focuses on that subject, emphasizing their functions in the environment and the use of antibody probes to study them. Proteins, 1995 Oct, 23(2), 241 - 55 Three-dimensional model of the alpha-subunit of bacterial luciferase; Sandalova T et al.; The predicted secondary structure of both subunits of bacterial luciferase is in accordance with a regular 8-fold alpha/beta-barrel structure . The 3D profile confirmed that luciferase subunits are compatible with the alpha/beta-barrel despite the absence of sequence similarity with any alpha/beta-barrel protein . The three-dimensional structure of 260 residues of the alpha-chain of luciferase was modeled from coordinates of glycolate oxidase and then energy minimized . The model obtained satisfies the criteria for the structure of a globular protein and is in accordance with known experimental data . From the model it is possible to predict active site residues involved in binding and catalysis . These predictions, and thus also the model, can be tested by protein engineering experiments. FEMS Immunol Med Microbiol, 1995 Oct, 12(2), 137 - 42 Localization of viable bacteria and bacterial antigens in arthritic joints of Erysipelothrix rhusiopathiae-infected pigs; Franz B et al.; Chronic polyarthritis was induced in pigs by infection with Erysipelothrix rhusiopathiae (serovar 2, strain T28) . Viable bacteria could be reisolated as long as 5 months post-infection from synovial fluid, synovial tissue and from isolated chondrocytes . The number of viable bacteria could be increased by hypotonic shock of the chondrocytes indicating a substantial intracellular amount of bacteria . Bacterial antigens were shown by immunohistochemistry to be present on the surface of both chondrocytes and synovial cells in arthritic joints . Neither viable bacteria nor bacterial antigen were detected in unaffected joints. Vaccine, 1995 Oct, 13(14), 1294 - 9 Gliding bacterial adjuvant stimulates feline cytokines in vitro and antigen-specific IgG in vivo; Zeidner NS et al.; Gliding bacterial adjuvant (GBA) has been previously characterized as a potent immune modulator, stimulating the growth of murine B lymphocytes, inducing murine NK cell activity, and promoting the release of several murine cytokines . Based on these studies and our interest in potentiating the effectiveness of feline vaccines, GBA was tested for its ability to stimulate feline T cells in vitro and act as a vaccine adjuvant in vivo . In vitro, GBA stimulated feline PBLs to proliferate and release interferon (IFN) and IL-2 . Unlike IFN, the release of IL-2 appeared to be unaffected by prior depletion of macrophages, indicating GBA directly stimulated feline T cells . In vivo GBA was co-administered with Keyhole Limpet Hemacyanin (KLH) and the anti-KLH antibody response was compared to cats receiving KLH emulsified in complete Freund's adjuvant (CFA) . Fourteen days after the third immunization and continuing for a 30-day observation period, KLH-specific IgG titers in cats receiving GBA were significantly higher than those given CFA . However, when cats were subsequently boosted with KLH alone, those cats receiving CFA demonstrated significantly higher antibody titers throughout a second 30-day observation period . The anti-KLH antibody memory response was greatly enhanced when GBA was emulsified with incomplete Freunds adjuvant (IFA) prior to injection . Serum titers of cats given KLH in an oil-based GBA preparation were significantly higher than cats receiving KLH adjuvanted with IFA or CFA, an effect which persisted 38 days after boosting with KLH alone . Finally, GBA significantly enhanced the feline humoral response to a recombinant protein of Dirofilaria immitis, the causative agent of feline heartworm . Serum titers of cats inoculated with recombinant antigen in GBA were significantly greater than cats given recombinant antigen adjuvanted with Titermax, alum, or NAGO . These studies indicate that GBA induces T cell proliferation and the release of IL-2 and IFN in vitro and can be used to enhance the recall antibody response to both a T cell dependent antigen and an immunogen derived from Dirofilaria immitis. Vet Immunol Immunopathol, 1995 Oct, 48(3-4), 287 - 98 Production and characterisation of ovine GM-CSF expressed in mammalian and bacterial cells; O'Brien PM et al.; A cDNA encoding ovine granulocyte-macrophage colony-stimulating factor (GM-CSF) was isolated and two forms of recombinant ovine GM-CSF were produced . A glycosylated form was produced in mammalian cells infected with a recombinant vaccinia virus encoding ovine GM-CSF . Recombinant ovine GM-CSF was also produced in Escherichia coli and purified by affinity chromatography . Both forms of the protein were detected by ovine GM-CSF-specific monoclonal antibodies, and exhibited activity on ovine bone marrow haemopoetic progenitor cells. Pharmacol Biochem Behav, 1995 Oct, 52(2), 347 - 54 Paradoxical conditioning of the plasma copper and corticosterone responses to bacterial endotoxin; Exton MS et al.; The cascade of physiologic mechanisms in response to infection, the acute phase response, is recognized as having a major role in host defense . Two such responses are an increase in plasma copper and activation of the hypothalamic-pituitary-adrenal axis, which are consistently reported to occur during bacterial infection . We aimed to determine whether the alterations in plasma copper and corticosterone were conditionable using the conditioned taste aversion paradigm . The regime involved the pairing of a novel-tasting saccharine solution (the conditioned stimulus) with lipopolysaccharide (the unconditioned stimulus) . Seven days after the initial pairing of these stimuli (the test day), the saccharine solution was represented . Animals exposed to this condition displayed a significant decrease in plasma copper levels . In addition, these rats experienced a reduction in plasma corticosterone that was time dependent . Paradoxically, the conditioned response of both these variables were in a direction contrary to that reported during bacterial infection . These results suggest that some acute phase responses may condition as a rebound response, or in an opposing trend to that occurring as the initial reaction. Genet Anal, 1995 Oct, 12(2), 73 - 9 Characterization of a human chromosome 22 enriched bacterial artificial chromosome sublibrary; Kim UJ et al.; Selection of chromosomal sublibraries from total human genomic libraries is critical for chromosome-based physical mapping approaches . We have previously reported a method of screening total human genomic library using flow sorted chromosomal DNA as a hybridization probe and selection of a human chromosome 22-enriched sublibrary from a total human bacterial artificial chromosome (BAC) library (Nucleic Acids Res 1995; 23: 1838-39) . We describe here further details of the method of construction as well as characterization of the chromosome 22-enriched sublibrary thus constructed . Nearly 40% of the BAC clones that have been mapped by fluorescence in situ hybridization (FISH) analysis were localized to chromosome 22 . By screening the sublibrary using chromosome 22-specific hybridization probes, we estimated that the sublibrary represents at least 2.5 x coverage of chromosome 22 . This is in good agreement with the results from FISH mapping experiments . FISH map data also indicate that chromosome 22-specific BACs in the sublibrary represent all the subregions of chromosome 22. Mol Biol Cell, 1995 Oct, 6(10), 1367 - 80 Computer analysis of the binding reactions leading to a transmembrane receptor-linked multiprotein complex involved in bacterial chemotaxis; Bray D et al.; The chemotactic response of bacteria is mediated by complexes containing two molecules each of a transmembrane receptor and the intracellular signaling proteins CheA and CheW . Mutants in which one or the other of the proteins of this complex are absent, inactive, or expressed at elevated amounts show altered chemotactic behavior and the phenotypes are difficult to interpret for some overexpression mutants . We have examined the possibility that these unexpected phenotypes might arise from the binding steps that lead to active complex formation . A limited genetic algorithm was used to search for sets of binding reactions and associated binding constants expected to give mutant phenotypes in accord with experimental data . Different sets of binding equilibria and different assumptions about the activity of particular receptor complexes were tried . Computer analysis demonstrated that it is possible to obtain sets of binding equilibria consistent with the observed phenotypes and provided a simple explanation for these phenotypes in terms of the distribution of active and inactive complexes formed under various conditions . Optimization methods of this kind offer a unique way to analyze reactions taking place inside living cells based on behavioral data. Lupus, 1995 Oct, 4(5), 339 - 47 Enhancement of renal disease in BXSB lupus prone mice after prior exposure to bacterial lipopolysaccharide; Granholm NA et al.; The mechanism, or mechanisms, responsible for enhancement of renal disease after episodes of infection are poorly understood . We used the BXSB mouse as a lupus model of autoimmune disease and we used bacterial lipopolysaccharide (LPS) as a surrogate infectious agent to gain some insight into the mechanism by which infections promote enhancement of autoimmune disease to chronicity . BXSB mice were exposed to LPS for 5 weeks, LPS was withdrawn and various tests and measurements were performed 6 weeks thereafter . Matched BXSB mice exposed to vehicle injections for 5 weeks served as controls . We verified that previous exposure to LPS enhances polyclonal B cell activation, impairs carrier function of blood cells for immune complexes, increases deposition of immune complexes in the microcirculation and promotes glomerular inflammation and sclerosis . These changes occurred at 6 weeks after withdrawal of LPS in the presence of unimpaired function of mononuclear phagocytes . Some of the effects of LPS are reversible, others are partially so and others are irreversible . Altered immune functions elicited by prior exposure to LPS can result in enhanced involvement of various renal compartments and can result in renal insufficiency. J Neuroendocrinol, 1995 Oct, 7(10), 791 - 9 Bacterial lipopolysaccharide-induced changes in FOS protein expression in the rat brain: correlation with thermoregulatory changes and plasma corticosterone; Hare AS et al.; In the present study the regions of the brain showing an increase in the number of FOS protein stained cells 180 min following intravenous saline or bacterial lipopolysaccharide (LPS) treatment were investigated and correlated with changes in body temperature and plasma corticosterone levels . Particular attention was given to the possible involvement of the circumventricular organs and regions of the brainstem containing central noradrenergic neurones . LPS at doses of 0.35, 3.5 and 50 micrograms caused highly significant increases in FOS protein expression in the organum vasculosum of lamina terminalis, the area postrema and the subfornical organ compared with saline controls . Marked increases in bacterial lipopolysaccharide-induced FOS protein expression were observed in the ventrolateral medulla, the nucleus of the solitary tract and the locus coeruleus which contain the A1, A2 and A6 noradrenergic neurones respectively . The changes in body temperature induced by LPS were found to be dependent upon the dose of LPS administered; the lowest dose employed (0.35 micrograms) induced an immediate and sustained fever, 3.5 micrograms LPS caused a biphasic response consisting of a hypothermic response followed by a febrile response, whereas 50 micrograms LPS induced a hypothermic response which then normalised by 160 min post-injection . Intravenous saline injection had no significant effect on body temperature . The occurance of LPS-induced hypothermia was coincident with increased FOS expression in the bed nucleus of stria terminalis, which houses vasopressinergic neurones involved in antipyresis, whereas in animals showing an LPS-induced febrile response there was no significant difference in the number of FOS stained cells in the bed nucleus of stria terminalis compared with saline treated animals . LPS also caused marked increases in FOS protein expression in the parvocellular regions of the paraventricular nucleus (pPVN) of the hypothalamus, the central nucleus of the amygdala and the ventral septal area . Plasma corticosterone was unaffected by the lowest dose of LPS (0.35 micrograms), however the higher doses employed (3.5 and 50 micrograms) caused significant increases in plasma corticosterone which correlated with the increases in the number of FOS stained cells in the pPVN . The results of the present study suggest that, in addition to the organum vasculosum of lamina terminalis, the area postrema and subfornical organ may be important in the responses to antigenic challenge that are mediated by the central nervous system . They also add support to the possible involvement of the bed nucleus of stria terminalis in LPS-induced hypothermia and of the involvement of the of the major noradrenergic cell groups (A1, A2 & A6) and a number of hypothalamic and extrahypothalamic forebrain regions in the interaction of immune and central nervous systems. Harefuah, 1995 Oct, 129(7-8), 229 - 32, 296 {Serodiagnosis of atypical bacterial respiratory infections}; Ben-Yaakov M et al.; Since prevalence of antibodies to bacteria causing atypical respiratory infections in Israel is as yet unknown, a 5-year antibody prevalence study was performed . Seroreactivity to Chlamydia pneumoniae (TWAR), with titers > or = 1:16 by microimmunofluorescence assay (MIF) was detected in 725/1305 (55.5%) of patients . 47/1012 ((4.6%) of adult patients had MIF results indicating recent infection with TWAR, (IgG titers of > or = 1:512, and/or IgM titers of > or = 1:16, and/or seroconversion) . Antibody prevalence and titers were low in children aged 1-10 years, increased in teenagers, and peaked in adults and the elderly, in whom prevalence was up to 79% and mean geometric titer up to 1:163 . Unlike the consistency in TWAR antibody prevalence and serological evidence of recent infection during the study period, a significant decrease in those variables was observed for Chlamydia trachomatis during the first 3 study years . Antibodies to M . pneumoniae were detected in 53 and to Legionella sp . in 47 out of 763 patients . There was serological evidence of recent infection with M . pneumoniae in 10 (including 7 children) and with Legionellae in 8 . Improved diagnosis of atypical respiratory infection might be achieved by the combined use of these proposed serological procedures. Zhonghua Yi Xue Za Zhi (Taipei), 1995 Oct, 56(4), 239 - 43 Elevation of serum IL-6 levels in patients with acute bacterial infection; Chen YM et al.; BACKGROUND . Serum cytokine levels have been reported to elevate in acute bacterial infection, but the relationship of differential elevation in cytokine levels to patients' clinical parameters and prognosis remains controversial . The present study was designed to evaluate whether serum interleukin-1 alpha (IL-1 alpha) and IL-6 levels were raised in patient with acute bacterial infection, and were correlated with patients clinical parameters . METHODS . Thirty patients, aged from 20 to 91 years, calling our emergency room with clinical evidence of acute bacterial infection and marked leukocytosis, were enrolled in this study . Sera were collected immediately and analyzed for IL-1 alpha and IL-6 levels with Enzyme-Linked Immunosorbent Assay (ELISA) method . RESULTS . All patients with acute bacterial infection had measurable higher levels of serum IL-6 than normal volunteers . Patients with higher serum IL-6 level were more likely to have fever, though without statistical significance (p = 0.09) . Serum IL-6 levels did not correlate significantly with positive blood culture result, septic shock, or fatal outcome . Serum IL-1 alpha levels were below minimal detectable concentrations in all patients checked . CONCLUSIONS . Serum IL-6 levels were elevated in patients with acute bacterial infection, and were possibly associated with the occurrence of fever . IL-1 alpha played no obvious role as systemic effector molecule in acute bacterial infection in our study. J Periodontol, 1995 Oct, 66(10), 864 - 9 Bacterial colonization of the external and internal sulci and of cellulose membranes at time of retrieval; Novaes AB Jr et al.; Membrane exposure with bacterial contamination is often considered one of the main reasons for the lack of predictability in achieving complete regeneration of periodontal defects . Ten cellulose membranes, retrieved from 7 patients treated for Class II furcation lesions in lower molars with at least 4 mm of exposure at time of retrieval were studied . Contamination of exposed membranes was studied using SEM analysis of four surfaces of the membrane, upper external, lower external, upper internal, and lower internal surfaces . DNA probe analysis of three periodontopathic bacteria, Porphyromonas gingivalis, Prevotella intermedia, and Actinobacillus actinmycetemcomitans, was carried out for specimens collected from the external and internal sulci . The results suggest that bacterial contamination of the membranes could be controlled if proper pre- and postoperative care is followed, since significant amounts of any of the three periodontopathogenic bacteria studied were not found . The SEM analysis corroborated the DNA probe analysis since the predominant morphotypes detected were not suggestive of periodontopathogenic bacteria . The importance of membrane contamination and of root concavities in the lack of predictability of the GTR procedure is discussed. Mol Ecol, 1995 Oct, 4(5), 627 - 31 DNA-based monitoring of total bacterial community structure in environmental samples; Holben WE et al.; Determining the structure of bacterial communities and their response to stimuli is key to understanding community function and the interactions that occur between micro-organisms and the environment . However, bacterial communities often comprise complex assemblages of large numbers of different bacterial populations . An approach is presented which allows bacterial community structure to be determined by fractionation of the complex mixture of total bacterial community DNA using the DNA-binding dye bisbenzimidazole which imposes G+C-dependent changes in the buoyant density of DNA . Bacterial community structure presented as percentage of total DNA vs . percentage G+C content of DNA is an indication of the relative abundance of phylogenetic groups of bacteria . Changes in the composition of a soil bacterial community in response to perturbations in the form of carbon amendment and altered water status were monitored. Arthritis Rheum, 1995 Oct, 38(10), 1513 - 8 Lack of detection of enteroviral RNA or bacterial DNA in magnetic resonance imaging-directed muscle biopsies from twenty children with active untreated juvenile dermatomyositis; Pachman LM et al.; OBJECTIVE . To investigate for the presence of increased titers of circulating antibody to putative infectious agents and for detectable viral RNA or bacterial DNA in children with active recent-onset juvenile dermatomyositis (DM) . METHODS . Magnetic resonance imaging-directed muscle biopsies were performed in 20 children with active, untreated, recent-onset juvenile DM and in age-matched children with neurologic disease . Sera were tested for complement-fixing antibody to Coxsackievirus B (CVB), influenza A and B, parainfluenza 1 and 3, Mycoplasma pneumoniae, mumps, respiratory syncytial virus, and Reovirus; and by immunofluorescence for IgG antibody to Toxoplasma gondii cytomegalovirus and IgM antibody to Epstein-Barr virus . Muscle from juvenile DM patients and control children, CD-1 Swiss mice with and without CVB1 infection, and viral stock positive for CVB1-6 were tested using reverse-transcriptase polymerase chain reaction with 5 primer sets, 4 probes (1 Coxsackievirus, 3 Enterovirus), and universal primers for DNA . RESULTS . No increased antibody, viral RNA, or bacterial DNA was present in the juvenile DM patients or the control children . CONCLUSION . Juvenile DM may be triggered by unidentified agent(s) in the genetically susceptible host. J Med Microbiol, 1995 Oct, 43(4), 251 - 7 In-vivo induction of apoptosis in murine lymphocytes by bacterial lipopolysaccharides; Norimatsu M et al.; The effect of bacterial lipopolysaccharide (LPS) on the lymphoid organs in C3H/HeN and C3H/HeJ mice was investigated . In C3H/HeN mice, LPS induced apoptosis, characterised by morphological nuclear condensation and DNA fragmentation resulting in thymic atrophy . Similar but less severe changes were also observed in the spleen and lymph nodes . In C3H/HeJ mice, only a slight depletion of lymphocyte numbers was observed in the lymphoid organs . The plasma endotoxin levels were dependent on the LPS dose regardless of mouse strain . On the other hand, the plasma TNF-alpha levels were significantly elevated in C3H/HeN mice 1 h post-injection and the time course of plasma corticosterone concentration correlated well with the development of apoptosis . These findings suggest that TNF-alpha and corticosterone may play an important role in LPS-induced apoptosis of lymphocytes. Cell Immunol, 1995 Oct 1, 165(1), 7 - 11 Effects of cyclosporin A and FK506 on nitric oxide and tetrahydrobiopterin synthesis in bacterial lipopolysaccharide-treated J774 macrophages; Hattori Y et al.; We evaluated the effects of the immunosuppressants cyclosporin A (CsA) and FK506 on the synthesis of nitric oxide (NO) induced by bacterial lipopolysaccharide (LPS) in J774 macrophages . CsA and FK506 each inhibited NO production by LPS in a concentration-dependent fashion, but the cytotoxicity was also evident at higher concentrations (100 microM) . Neither CsA nor FK506 had any effect on the activity of NO synthase (NOS) that had already been induced . Findings indicated that CsA and FK506 inhibit the induction of NOS, rather than its catalytic activity . CsA and, to a lesser extent, FK506 increased the synthesis of tetrahydrobiopterin (BH4), an essential cofactor of NOS . Thus, inhibition of NO formation by CsA or FK506 is unlikely to associated with a change in BH4 synthesis caused by these agents in LPS-treated J774 macrophages. Leukemia, 1995 Oct, 9 Suppl 1, S61 - 3 Hematological reconstitution and gene therapy: retroviral transfer of the bacterial beta-galactosidase activity into human hematopoietic CD34+ cell populations and into T lymphocytes derived from the peripheral blood; Bagnis C et al.; We report the possibility to transfer marker genes coding for beta-galactosidase activity using retroviral vectors into human peripheral blood CD34+ cells, peripheral blood T-lymphocytes and into the growth factor-dependent human hematopoietic cell line TF-1 . Using the MFG-nisLacZ and the FLac vector and various packaging cell lines, we demonstrated retroviral transfer and high expression of a bacterial beta-galactosidase activity induced by the nisLacZ gene or the Sh-ble/LacZ gene . Kinetics of expression of the transgenes were analyzed both in primary cells and cell lines . Absence of cytotoxicity related to the expression of the bacterial beta-galactosidase was assessed in both cell types . These results open interesting prospectives for the use of the beta-galactosidase activity to mark and follow the fate of genetically modified cells isolated from patients prior to reimplantation. N Engl J Med, 1995 Sep 28, 333(13), 845 - 51 Bacterial pneumonia in persons infected with the human immunodeficiency virus . Pulmonary Complications of HIV Infection Study Group; Hirschtick RE et al.; BACKGROUND . Patients with human immunodeficiency virus (HIV) infection are at increased risk for bacterial pneumonia in addition to opportunistic infection . However, the risk factors for bacterial pneumonia and its incidence in this population are not well defined . METHODS . In a multicenter, prospective, observational study, we monitored 1130 HIV-positive and 167 HIV-negative participating adults for up to 64 months for pulmonary disease . The HIV-positive group comprised 814 homosexual or bisexual men, 261 injection-drug users, and 55 female partners of HIV-infected men . RESULTS . There were 237 episodes of bacterial pneumonia among the HIV-positive participants (rate, 5.5 per 100 person-years), as compared with 6 episodes among the HIV-negative participants (rate, 0.9 per 100 person-years; P < 0.001) . The rate of bacterial pneumonia increased with decreasing CD4 lymphocyte counts (2.3, 6.8, and 10.8 episodes per 100 person-years in the strata with more than 500, 200 to 500, and fewer than 200 cells per cubic millimeter, respectively; P < or = 0.022 for each comparison) . Injection-drug users had a higher rate of bacterial pneumonia than did homosexual or bisexual men or female partners . In the stratum with the fewest CD4 lymphocytes, cigarette smoking was associated with an increased rate of pneumonia . Mortality was almost four times higher among participants with an episode of pneumonia than among the others . Prophylaxis with trimethoprim-sulfamethoxazole was associated with a 67 percent reduction in confirmed episodes of bacterial pneumonia (P = 0.007) . CONCLUSIONS . Bacterial pneumonia is more frequent in HIV-positive persons than in seronegative controls, and the risk is highest among those with CD4 lymphocyte counts below 200 per cubic millimeter and among injection-drug users. Nucleic Acids Res, 1995 Sep 25, 23(18), 3673 - 7 Identification of the yeast nuclear gene for the mitochondrial homologue of bacterial ribosomal protein L16; Pan C et al.; An open reading frame encoding a member of the L16 family of ribosomal proteins is adjacent to the URA7 gene on the left arm of chromosome II in Saccharomyces cerevisiae . The predicted L16-like polypeptide is basic (pl 11.12), contains 232 amino acids (26.52 kDa) and has 36% amino acid sequence identity to E . coli L16 . Immunoblot analysis with polyclonal antibodies to the L16-like polypeptide showed specific cross-reaction with a 22,000 Mr mitochondrial polypeptide that co-sediments with the large subunit of the mitochondrial ribosome in sucrose density gradients . The levels of the L16 mRNA and protein varied in response to carbon source . In {rho degree} cells lacking mitochondrial rRNA, the L16 mRNA accumulated at normal levels, but the protein was barely detectable, indicating RNA-dependent accumulation of the L16 protein . Gene disruption experiments demonstrated that the yeast mitochondrial L16 is an essential ribosomal protein in vivo. Genomics, 1995 Sep 20, 29(2), 413 - 25 Construction and characterization of a bovine bacterial artificial chromosome library; Cai L et al.; A bacterial artificial chromosome (BAC) library has been constructed for use in bovine genome mapping using constructed for use in bovine genome mapping using the pBeloBAC11 vector . Currently, the library consists of 23,040 clones, which achieves a 70% probability (P=0.70) of the library containing a specific unique DNA sequence . Sixty thousand clones, or about three haploid bovine genomes, will be required to achieve a 95% probability (P=0.95) of containing a unique sequence . An average insert size of 146 kb was estimated from the analysis of 77 randomly selected BAC clones produced by one or two rounds of size selection . The bovine DNA inserts proved to be very stable for at least 100 cell generations . No chimeric clones were detected among 11 large, size-selected BAC clones using fluorescence in situ hybridization (FISH) on metaphase bovine chromosomes . Thirty-three of 46 (72%) sequences were present in the library in at least one copy, which is consistent with the estimated 70% probability of this library containing a unique DNA sequence . A BAC clone as sequence-tagged sites for genetic mapping . These markers cosegregated, and no recombinants were detected in 193 informative meioses . Plasmid end rescue and the inverse polymerase chain reaction methods were used to rescue both ends of this BAC clone, and chromosome walking was performed using PCR primers designed within the end region sequences . Based on our experimental results, the BAC system provides a very useful tool for complex genome analysis. Genes Dev, 1995 Sep 15, 9(18), 2305 - 13 A novel bacterial transcription cycle involving sigma 54; Tintut Y et al.; sigma 54 is the promoter recognition subunit of the form of bacterial RNA polymerase that transcribes from promoters with enhancer elements . DNase footprinting experiments show that sigma 54 is attached selectively to the template strand, which must be single-stranded for transcription initiation . sigma 54 remains bound at the promoter after core polymerase begins elongation, in contrast to the well-established sigma 70-holoenzyme transcription cycle . Permanganate footprinting experiments show that the bound sigma 54 and the elongating core RNA polymerase downstream of it are each associated with a single-strand DNA region . Template commitment assays show that the promoter-bound sigma 54 must be reconfigured before reinitiation of transcription can occur . This unexpected pathway raises interesting possibilities for transcriptional regulation, especially with regard to control at the level of reinitiation. FEMS Microbiol Lett, 1995 Sep 15, 131(3), 235 - 42 Structure and partitioning of bacterial DNA: determined by a balance of compaction and expansion forces? Woldringh CL, Jensen PR, Westerhoff HV. The mechanisms that determine chromosome structure and chromosome partitioning in bacteria are largely unknown . Here we discuss two hypotheses: (i) the structure of the Escherichia coli nucleoid is determined by DNA binding proteins and DNA supercoiling, representing a compaction force on the one hand, and by the coupled transcription/translation/translocation of plasma membrane and cell wall proteins, representing an expansion force on the other hand; (ii) the two forces are important for the partitioning process of chromosomes. Biochemistry, 1995 Sep 12, 34(36), 11606 - 16 Binding sites of quinones in photosynthetic bacterial reaction centers investigated by light-induced FTIR difference spectroscopy: symmetry of the carbonyl interactions and close equivalence of the QB vibrations in Rhodobacter sphaeroides and Rhodopseudomonas viridis probed by isotope labeling; Breton J et al.; The photoreduction of the secondary quinone acceptor QB in reaction centers (RCs) of the photosynthetic bacteria Rhodobacter sphaeroides and Rhodopseudomonas viridis has been investigated by light-induced FTIR difference spectroscopy of RCs reconstituted with several isotopically labeled ubiquinones . The labels used were 18O on both carbonyls and 13C either uniformly or selectively at the 1- or the 4-position, i.e., on either one of the two carbonyls . The QB-/QB spectra of RCs reconstituted with the isotopically labeled and unlabeled quinones as well as the double differences calculated from these spectra exhibit distinct isotopic shifts for a number of bands attributed to vibrations of QB and QB- . The vibrational modes of the quinone in the QB site are compared to those of ubiquinone in vitro, leading to band assignments for the C = O and C = C vibrations of the neutral QB and for the C***O and C***C of the semiquinone . The C = O frequency of each of the carbonyls of the unlabeled quinone is revealed at 1641 cm-1 for both species . This demonstrates symmetrical and weak hydrogen bonding of the two C = O groups to the protein at the QB site . In contrast, the C = C vibrations are not equivalent for selective labeling at C1 or at C4, although they both contribute to the approximately 1617-cm-1 band in the QB-/QB spectra of the two species . Compared to the vibrations of isolated ubiquinone, the C = C mode of QB does not involve displacement of the C4 carbon atom, while the motion of C1 is not hindered . Further analysis of the the spectra suggests that the protein at the binding site imposes a specific constraint on the methoxy and/or the methyl group proximal to the C4 carbonyl . The FTIR observations provide compelling evidence for almost identical conformation and identical interactions of the ubiquinone in the QB binding site of Rb . sphaeroides and Rp . viridis in contrast to the X-ray structures, which yield different descriptions for the hydrogen-bonding pattern of QB binding . In the semiquinone state, the bonding interactions of the C***O groups are also symmetrical and the C***C are inequivalent at C1 and C4 . However, the interactions are almost the same in the RCs of both species. Nucleic Acids Res, 1995 Sep 11, 23(17), 3554 - 62 Detection of new genes in a bacterial genome using Markov models for three gene classes; Borodovsky M et al.; We further investigated the statistical features of the three classes of Escherichia coli genes that have been previously delineated by factorial correspondence analysis and dynamic clustering methods . A phased Markov model for a nucleotide sequence of each gene class was developed and employed for gene prediction using the GeneMark program . The protein-coding region prediction accuracy was determined for class-specific Markov models of different orders when the programs implementing these models were applied to gene sequences from the same or other classes . It is shown that at least two training sets and two program versions derived for different classes of E . coli genes are necessary in order to achieve a high accuracy of coding region prediction for uncharacterized sequences . Some annotated E . coli genes from Class I and Class III are shown to be spurious, whereas many open reading frames (ORFs) that have not been annotated in GenBank as genes are predicted to encode proteins . The amino acid sequences of the putative products of these ORFs initially did not show similarity to already known proteins . However, conserved regions have been identified in several of them by screening the latest entries in protein sequence databases and applying methods for motif search, while some other of these new genes have been identified in independent experiments. FEBS Lett, 1995 Sep 11, 371(3), 303 - 6 Interaction of the bacterial protein toxin alpha-haemolysin with model membranes: protein binding does not always lead to lytic activity; Ostolaza H et al.; alpha-Haemolysin interaction with model membranes has been investigated by a 2-fold procedure . First, protein binding has been measured, by a direct method as well as through changes in the intrinsic fluorescence of the protein when incubated with liposomes and divalent cations . Then, the above results have been correlated with the protein lytic activity . The extent of protein binding is not significantly modified by the presence or absence of Ca2+, or by changes in lipid composition, although these factors influence greatly the membrane lytic activity of the protein . Moreover, Ca2+ binding to the toxin must occur prior to protein binding to the bilayer, for a lytic effect to take place. Biochemistry, 1995 Sep 5, 34(35), 11176 - 85 Effects of bacterial endotoxin on human cross-linked and native hemoglobins; Kaca W et al.; Previous investigations have demonstrated that hemoglobin (Hb) is a binding protein for bacterial endotoxin (lipopolysaccharide, LPS) and that the structure and biological activity of LPS are altered in the presence of Hb . In the present study, the influence of LPS on the structure of native human HbA0 and covalently cross-linked Hb (alpha alpha Hb) was studied by analyzing the absorption and circular dichroic spectra of Hb in the wavelength region of 200-650 nm . Incubation of oxyHb with each of several LPSs resulted in a decrease in the intensity of the major Soret band at 414 nm with a shift in the maximum peak to 410 nm, decreases in the intensities of the major visible region peaks at 541 and 577 nm, and the appearance of increased absorbance in the visible region in the range of 630 nm . The resultant spectra are characteristic of methemoglobin formation . These spectral changes were time-dependent and LPS-concentration-dependent . Production of methemoglobin was prominent with chemically modified, partially deacetylated rough LPS, and was observed to a lesser extent both with native, complete rough and with native smooth LPSs . The influence of LPS on the absorption spectrum of methemoglobin also was directly tested . The conversion of methemoglobin to hemichrome in the presence of LPS was demonstrated and was shown to be reversible . Analysis of circular dichroic spectra of Hb demonstrated LPS-induced spectral changes in the visible and Soret regions consistent with the production of a substantial quantity of metHb, but did not demonstrate any alteration in the far-UV region (210-240 nm) . Moreover, Hb oxygen affinity was only slightly altered after incubation with any of several LPSs . In conclusion, analyses of absorption and circular dichroic spectra reveal the potential of LPS to produce a facilitated oxidation of both alpha alpha-cross-linked human Hb and native human HbA0, without substantial changes in the secondary structure of the globin. Biochemistry, 1995 Sep 5, 34(35), 10970 - 5 Secondary structure and stability of the bacterial carbohydrate-specific recognition proteins K88ab, AFA-1, NFA-1, and CFA-1; Knorle R et al.; The conformation and thermodynamic stability of the four polymeric carbohydrate-specific bacterial recognition proteins K88ab, AFA-1, NFA-1, and CFA-1 and their monomeric subunits that can be obtained by variation of pH were studied by infrared spectroscopy and differential scanning microcalorimetry . For NFA-1, a pH-dependent dissociation of the polymeric form cannot be achieved due to the stronger interactions of the neighboring subunits . Generally, no alterations in secondary structure are observed between the monomeric and the polymeric proteins . All adhesins reveal a high degree of beta-sheet structure (40-55%), while the alpha-helix component is of minor importance (10-20%) . The adhesins investigated in this study revealed unusually high denaturation temperatures (69-104 degrees C) and stabilizing Gibbs energies, delta G (40-125 kJ/mol), compared to common globular proteins . Statistical deconvolution of the DSC curves yields a two-state transition of K88ab, NFA-1, and the monomeric CFA-1 and the existence of intermediate states for AFA-1 and polymeric CFA-1 during the denaturation process . The irreversible denaturation of K88ab, AFA-1, and CFA-1 is explained by aggregation of the polypeptide chains forming a three-dimensional network of intermolecular beta-sheet-type structures . In contrast, denaturation of NFA-1 is completely reversible . At a physiologically relevant temperature of approximately 40 degrees C, we observe predenaturational events in the DSC curves of polymeric K88ab and NFA-1 with no concomittant changes in the secondary structure of these proteins. Indian Pediatr, 1995 Sep, 32(9), 989 - 93 Cranial sonography in bacterial meningitis; Mahajan R et al.; Cranial ultrasonography was performed on 61 infants with acute bacterial meningitis (ABM) . Thirty nine, infants (64%) had acute meningitis with no clinical evidence of complications (Group-I) and 22 infants (36%) had clinical evidence of complications of ABM (Group-II) . Cranial ultrasound was normal in 20 infants (32.8%) . The spectrum of sonographic abnormalities included echogenic sulci (60.6%), sulcal separation (49.8%), abnormal parenchymal echoes (42.6%), ventriculomegaly (34.4%), ventriculitis (19.7%), abscess (3.3%), subdural empyema (1.63%) and hemorrhagic infarct (1.63%) . Various abnormal findings were seen in all 22 patients of Group II (100%) and in only 19 out of 39 patients in Group I (31.9%) . Cranial sonography was comparable to CT scan done in 10 cases of Group II . Our study suggests that ultrasound is a quick, reliable and effective diagnostic tool in diagnosis and management of infants with or without evidence of complications. Protein Eng, 1995 Sep, 8(9), 859 - 63 Bacterial secretion of the Fab fragment of a mouse monoclonal IgM that reacts with IgG variable regions; Adib-Conquy M et al.; In this report, we describe the expression system that enabled us to produce in Escherichia coli the Fab fragment of a mouse IgM that has previously been shown to inhibit the binding of IgG to autoantigens by interacting with their variable regions . In our system, both light chain and heavy chain fragments were put under the control of the malE promoter . The light chain was fused to the MalE signal sequence, while the heavy chain variable and first constant region were fused to the alkaline phosphatase signal sequence . In this system, after induction of the promoter with maltose, the Fab fragment could be detected in a periplasmic extract of the bacteria by Western blotting and also by ELISA . This Fab fragment was purified on a goat anti-mouse immunoglobulin immunoadsorbent and biotinylated . The Fab fragment produced by E.coli reacted with the trinitrophenyl (TNP) hapten and F(ab')2 fragments of mouse IgG and these reactivities could be specifically inhibited by the corresponding soluble antigens . The dissociation constants of this Fab were 1.65 x 10(-6) M for TNP and 5 x 10(-6) M for IgG F(ab')2 fragments, indicating that the affinity of the Fab fragment compared with that of the whole IgM molecule was similar for TNP but was lower for IgG F(ab')2 fragments. Vet Pathol, 1995 Sep, 32(5), 489 - 97 An animal model of gastric ulcer due to bacterial gastritis in mice; Eaton KA et al.; Conventional female BalbC mice were inoculated with Gastrospirillum-like bacteria in mouse gastric homogenate or in 5.0-microns filtrate of gastric homogenate . The bacteria were originally isolated from cheetahs with gastritis . The mice were killed 6 months, 7 months, or 1 year after inoculation . All mice became infected with Gastrospirillum-like bacteria that were confined to the gastric mucosa . Control mice, given either sterile Brucella broth, 0.22-microns filtrate of infected gastric homogenate, or uninfected gastric homogenate did not become infected with bacteria . Lesions in infected mice included severe lymphoplasmacytic gastritis (26/26 infected mice), gastric epithelial hyperplasia (25/26 infected mice), and gastric ulceration (11/26 infected mice) . Neutrophilic inflammatory cell infiltrates were inconsistent . None of the uninfected control mice had Gastrospirillum-like bacteria, gastritis, gastric epithelial hyperplasia, or gastric ulceration . These results implicate Gastrospirillum-like bacteria from cheetahs in the pathogenesis of gastric ulceration . This model will be useful in investigating the mechanisms of gastric ulceration associated with bacterial gastritis. Drug Metab Dispos, 1995 Sep, 23(9), 945 - 50 Bacterial expression and kinetic characterization of the human monoamine-sulfating form of phenol sulfotransferase; Ganguly TC et al.; The cDNA for the human monoamine-sulfating form of phenol sulfotransferase (hM-PST) was isolated from a T47D human breast carcinoma lambda gt10 cDNA library, and the active enzyme was expressed in Escherichia coli . Expressed hM-PST was very similar to the brain, intestinal, and platelet forms of the enzyme in its physical, immunological, and kinetic properties . The ability of hM-PST to sulfate a number of xenobiotics was examined and compared with the bacterially expressed human phenol-sulfating form of PST (hP-PST) . The translation product of the T47D hM-PST cDNA was 92% identical to that of liver hP-PST . Monoamine neurotransmittors, such as epinephrine and dopamine, were maximally conjugated at lower concentrations by expressed hM-PST (2 and 20 microM, respectively) than by hP-PST (1 and 1 mM, respectively) . In contrast, simple phenols--such as p-nitrophenol, acetaminophen, and alpha-naphthol--were maximally conjugated at lower concentrations (4 microM, 20 microM, and 0.5 microM, respectively) by hP-PST than by hM-PST (1 mM, 1.5 mM, and 50 microM, respectively) . Minoxidil was sulfated at similar rates and concentrations (7 mM) by both forms of PST . None of the estrogens or related compounds, such as beta-estradiol, 17 alpha-ethinylestradiol, diethylstilbestrol, equilenin, or genistein tested as substrates were sulfated by hM-PST; however, all of these compounds were substrates for hP-PST . As with hP-PST, the hydroxysteroids dehydroepiandrosterone and cortisol were not sulfated by hM-PST.(ABSTRACT TRUNCATED AT 250 WORDS) Mutagenesis, 1995 Sep, 10(5), 447 - 8 The use of a streamlined bacterial mutagenicity assay, the MINISCREEN; Brooks TM; A streamlined bacterial mutagenicity assay, the MINISCREEN, was developed to enable the rapid screening of a large number of chemical compounds on a purely qualitative basis . Experiments with a series of known carcinogens/mutagens and non-carcinogens/mutagens showed a good correlation with conventional bacterial mutagenicity assays (Ames tests), and the subsequent testing of over 300 candidate agricultural chemicals and over 100 industrial chemicals has proven its value as a screening method. Protein Sci, 1995 Sep, 4(9), 1768 - 79 Structure and function of microplasminogen: II . Determinants of activation by urokinase and by the bacterial activator streptokinase; Wang J et al.; We have used a group of human microplasminogens (mPlg), modified by residue substitutions, insertions, deletions, and chain breaks (1) to study the determinants of productive interactions with two plasminogen activators, urokinase (uPA), and streptokinase (SK); (2) to explore the basis of species specificity in the zymogen-SK complex activity; and (3) to compare active SK complex formation in mPlg and microplasmin (mPlm) . Modifications within the disulfide-bonded loop containing the activation site and the adjacent hexadecapeptide upstream sequence showed that uPA recognition elements encompassed R29 at the activation site and multiple elements extending upstream to perhaps 13 residues, all maintained in specific conformational register by surrounding pairs of disulfide bonds . A generally parallel pattern of structural requirements was observed for active zymogen-SK complex formation . Changes within the loop downstream of the activation site were tolerated well by uPA and poorly by SK . The introduction of selected short bovine (Plg) sequences in human mPlg reduced the activity of the resulting SK complexes . The requirements for active SK complex formation are different for mPlg and mPlm. AIDS, 1995 Sep, 9(9), 1093 - 7 Bacterial vaginosis and HIV seroprevalence among female commercial sex workers in Chiang Mai, Thailand; Cohen CR et al.; OBJECTIVE: To investigate the relationship between HIV seropositivity and bacterial vaginosis (BV) in a population at high risk for sexual acquisition of HIV . DESIGN: A cross-sectional study was conducted among 144 female commercial sex workers in Chiang Mai, Thailand . METHODS: The participants were tested for cervical gonorrhea and Chlamydia infection, syphilis, Trichomonas vaginitis, Candida vaginitis, BV, and HIV infection . BV was diagnosed by clinical criteria (pH > 4.5, positive amine test, and presence of clue cells) and using Gram stains . RESULTS: Thirty-three per cent of participants had BV, and 43% were HIV-positive . Using clinical criteria, the association of BV and HIV seropositivity was significant {odds ratio (OR), 2.7; 95% confidence interval (CI), 1.3-5.0} . Although the association between BV and HIV prevalence was not significant using Gram stains alone for diagnosis of BV, an association was found between abnormal vaginal flora and HIV (OR, 2.1; 95% CI, 1.0-4.8) . In multiple logistic regression analysis, adjusting for age, number of sexual encounters per week, current condom use, and currently having a sexually transmitted disease (STD), both BV and a history of an STD were independently associated with HIV seropositivity (adjusted OR for BV, 4.0 and 95% CI, 1.7-9.4; adjusted OR for history of an STD, 6.9 and 95% CI, 2.1-22.9) . CONCLUSIONS: When diagnosed clinically, BV is independently associated with HIV seroprevalence . HIV infection may promote abnormal vaginal flora, or BV may increase susceptibility to sexual transmission of HIV . Alternatively, the association seen here may result from intervening variables; in this case BV may be a marker or a cofactor of HIV transmissionPIP: To explore a possible association between bacterial vaginosis and human immunodeficiency virus (HIV) infection, 144 consecutively enrolled commercial sex workers from a sexually transmitted disease clinic (STD) in Chiang Mai, Thailand, were interviewed and underwent serologic testing and genital examination . 62 (43%) of sex workers were HIV-positive . A self-reported history of syphilis, chancroid, herpes, gonorrhea, or Chlamydia was significantly associated with HIV infection . Bacterial vaginosis, detected in 49 (34%), was also associated with HIV infection . Sex workers reporting 10-19 and 20 or more sexual encounters per week were 2.2 and 3.5 times, respectively, more likely to be infected with HIV than those reporting under 10 encounters . A clinically established diagnosis of bacterial vaginosis was independently associated with HIV seropositivity even when age, number of sexual encounters per week, current condom use, and past and current STD infection were controlled (odds ratio, 4.0; 95% confidence interval, 1.7-9.4) . When the bacterial vaginosis diagnosis was based on Gram stain (score 7-10), however, the association with HIV seropositivity disappeared, but having abnormal vaginal flora (gram stain score 4-10) was related to HIV status . Further epidemiologic studies are recommended to investigate the possibility that bacterial vaginosis--the most prevalent genital infection in Thailand--acts as a cofactor for the heterosexual transmission of HIV . Biosci Biotechnol Biochem, 1995 Sep, 59(9), 1790 - 2 Synthesis and mitogenic activity of nonreducing-sugar subunit analogs of bacterial lipid A composed only of 3-hydroxytetradecanoic acid and its homologs; Ishida H et al.; Novel analogs of the nonreducing-sugar subunit of bacterial lipid A, which were composed only of 3-hydroxytetradecanoic acid and its homologs, were synthesized . These analogs exhibited significant mitogenic activity. Biosci Biotechnol Biochem, 1995 Sep, 59(9), 1679 - 87 Two-dimensional gel electrophoresis of ribosomal proteins as a novel approach to bacterial taxonomy: application to the genus Arthrobacter; Ochiai K et al.; Ribosomal proteins from 22 strains of 15 different species belong to the genus Arthrobacter were analyzed by an improved two-dimensional gel electrophoresis . Electrophoretograms of ribosomal proteins from 15 type strains had species-specific patterns . Similarity coefficients (SAB values) of ribosomal proteins with mol . wt . of greater than about 20,000, among strains of the same species (DNA relatedness values of more than 61%) were greater than 0.85, but the SAB values among strains of different species were less than 0.60 . The N-terminal amino acid sequences of the AL2 proteins, which migrated into similar positions in this method, from 5 type strains were shown to be highly homologous . Our results indicated that ribosomal proteins have been conserved within species during evolution and that the members of the genus Arthrobacter are phylogenetically homogeneous . Thus, ribosomal protein profiles by this method are a potential tool for strain identification. Minerva Gastroenterol Dietol, 1995 Sep, 41(3), 227 - 30 {Diagnostic aspects of spontaneous bacterial peritonitis in hepatic cirrhosis with ascites}; Breda A et al.; Spontaneous bacterial peritonitis (SBP) is a severe complication appearing in 8-22% of hepatic cirrhosis with ascitic decompensation . The authors describe 6 cases of SBP in hepatic cirrhosis . SBP diagnosis has been confirmed by isolation of the aetiological agents in the ascitic fluid, and PMN count above 250/ml in 12% of the studies cases . SBP diffusion and the high mortality risk justify the examine of the ascitic fluid also in asymptomatic patients. Nutr Hosp, 1995 Sep-Oct, 10(5), 272 - 8 {Nutrition and bacterial translocation}; Lozano Sanchez F et al.; The present work is part of a presentation given at the Scientific Meeting of the Association for Surgical Nutrition and Metabolism, during the XX National Congress for Surgery (Madrid, November 1994) . The authors, prior to presenting their experiences, define and high light the importance of the phenomenon of "Bacterial Translocation" (BT) . Afterwards, and based on several experimental studies performed by them, they attempt to answer two questions: 1) Is the term BT correct? 2) Is BT a physiological or a pathological state? Finally they review the relationship which exists between bacterial translocation and nutrition, both from a causative point of view as from the prevention and therapy of the same. J Ind Microbiol, 1995 Sep, 15(3), 243 - 7 Bacterial adhesion measurements on soft contact lenses using a Modified Vortex Device and a Modified Robbins Device; Schultz CL et al.; S . marcescens 8100 and P . aeruginosa 15442 were used to study bacterial adhesion to hydrogel contact lenses which had not been worn . Bacterial removal from unworn lens materials was assessed with a calibrated vortex device modified with a digital rpm readout and fitted with a test tube attachment (MVD) . The MVD, which relies on a whirlpool-like force to remove the bacteria, showed that bacteria adhered to the same degree to etafilcon A, vifilcon A and polymacon lenses under standardized conditions . Tracking the isoenzyme patterns of these bacterial species over time showed instability of S . marcescens upon repeated passage . This instability was not evident with P . aeruginosa . Bacterial adhesion of P . aeruginosa 15442, to human worn and unworn etafilcon A materials was determined with a Modified Robbins Device . The MRD was closed off at both ends stopping medium and bacterial movement after 1 h of fluid flow over the lens surface . The results show that immediately following this 1-h period more bacteria adhere to unworn contact lenses than to worn lenses . However, bacterial counts were equivalent on worn and unworn lenses following 5 h of static incubation. Br J Cancer, 1995 Sep, 72(3), 634 - 6 High-dose interleukin 2 promotes bacterial translocation from the gut; Reynolds JV et al.; Toxicity associated with high-dose recombinant interleukin 2 (rIL-2) therapy simulates a sepsis syndrome, but the mechanism remains unclear . We hypothesised that translocated gut-origin bacteria may be important . Fifty-one male rats were randomised to receive rIL-2 by intraperitoneal injection at doses (IU) of 10(5) (n = 15), 10(4) (n = 8), 10(3) (n = 8) or 10(2) (n = 8) twice daily, or a saline bolus (n = 12) . After 5 days, ileal histomorphology was assessed and the mesenteric lymph node complex cultured . Results showed that colonisation of mesenteric lymph nodes with Escherichia coli occurred in all rats treated with 10(5) IU of rIL-2, and in 62%, 37% and 12% of rats treated with decreasing doses of rIL-2 . No translocation was observed in control animals . An increase in submucosal lymphatics and occasional mucosal disruption was seen only in the group receiving 10(5) IU . These data show that rIL-2 promotes bacterial translocation and suggests a mechanism that may fuel high-dose rIL-2 toxicity in man. Biochem J, 1995 Sep 1, 310 ( Pt 2), 709 - 14 Effect of adrenaline and phorbol myristate acetate or bacterial lipopolysaccharide on stimulation of pathways of macrophage glucose, glutamine and O2 metabolism . Evidence for cyclic AMP-dependent protein kinase mediated inhibition of glucose-6-phosphate dehydrogenase and activation of NADP+-dependent 'malic' enzyme; Costa Rosa LF et al.; Adrenaline has recently been shown to stimulate both glucose metabolism and H2O2 release by macrophages but the activity of the key pentose phosphate pathway enzyme, glucose-6-phosphate dehydrogenase (which generates the NADPH crucial for the reduction of molec