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| Cell, 1992 Apr 3, 69(1), 5 - 8 DNA replication, the bacterial cell cycle, and cell growth; Zyskind JW et al.; The coupling of replication to the cell cycle and cell growth involves events that occur at oriC . Immediately after initiation, there is an eclipse phase during which reinitiation from the newly synthesized origins is prevented . GATC sites in oriC remain in a hemimethylated state longer than other sites because of their association with the outer membrane, which prevents DnaA from binding and activating additional rounds of initiation . After the origins are methylated and released from the outer membrane, the concentration of newly synthesized DnaA and the activation of oriC by transcription from the nearby mioC and gid promoters determine when the next rounds of replication initiate . If growth rate is reduced, the synthesis of (p)ppGpp will increase, and this will lead to a decrease in dnaA, mioC, and gid transcription . On the other hand, if growth rate is increased by access to a tasty meal, synthesis of (p)ppGpp will decrease, expression of dnaA, mioC, and gid genes will increase, and a shortening of the interinitiation time will result . The participation of all these control features ensures rapid and precise coordination of DNA replication with cell growth. Thromb Haemost, 1992 Apr 2, 67(4), 428 - 33 Bacterial expression of biologically active high molecular weight kininogen light chain; Kunapuli SP et al.; Human high molecular weight kininogen (HK), a single chain plasma glycoprotein, serves as a cofactor in the contact system of blood coagulation . After cleavage by human plasma kallikrein, the nonapeptide bradykinin is released . The HK light chain (LC) contains coagulant activity, which requires both the ability to bind the contact system zymogens, prekallikrein and factor XI, and the ability to interact with negatively charged surfaces . Since bacterial expression might not be successful if carbohydrate was required for activity, we evaluated that possibility by incubating plasma HK with endoglycosydase F . Although the procedure removed detectable N-linked carbohydrate, no change in specific activity occurred . We then developed a bacterial expression system to produce recombinant HK LC . The cDNA coding for the HK LC was prepared by polymerase chain reaction (PCR), digested with restriction enzymes EcoRI and PstI, and introduced into the bacterial expression vector pKK223-3 . E . coli harboring this recombinant plasmid (pSK1) expressed HK LC upon induction with isopropylthio-galactoside (IPTG) . The recombinant protein (27 kDa), when transferred onto a PVDF membrane, was recognized by monospecific polyclonal anti-HK LC-antibodies . The recombinant HK LC was purified by heparin agarose affinity chromatography to homogeneity and found to have a specific activity of 28 coagulant units per mg protein, similar to the specific activity of the LC derived by proteolytic digestion of human plasma HK . We conclude: 1) The HK LC synthesized in bacteria is biologically active, and 2) the 40% carbohydrate content of the HK LC is not required for its cofactor activity. Singapore Med J, 1992 Apr, 33(2), 125 - 30 Clinical features and haematological indices of bacterial infections in young infants; Hiew TM et al.; The clinical features and haematologic indices of 100 young infants aged 3 months and below, admitted with suspected bacterial infections, were analysed . Fever, lethargy, hepatomegaly, poor feeding and irritability were the commonest features for suspecting a bacterial infection in these infants . However, the features significantly associated with bacterial infections were respiratory distress and cyanosis . Of the haematologic indices commonly associated with bacterial infections, only C-reactive protein and erythrocyte sedimentation rate were significantly predictive compared to leukocyte counts, absolute neutrophil counts and nitro-blue tetrazolium tests . When used in combination, a raised C-reactive protein with erythrocyte sedimentation rate, a raised erythrocyte sedimentation rate with abnormal leukocyte counts and a raised C-reactive protein with abnormal leukocyte counts were significantly associated with bacterial infections. Vet Immunol Immunopathol, 1992 Apr, 32(1-2), 139 - 48 The intestinal and serum humoral immune response of mice to orally administered antigens in liposomes: II . The response to liposome-entrapped bacterial proteins; Clarke CJ et al.; Effective oral adjuvants are needed to improve the intestinal immune responses to oral vaccines that are based on relatively low molecular weight antigens refined from veterinary pathogens . Liposomes prepared by different methods and composed of phospholipids of varying transition temperature were used to entrap cholera toxin (CT) and fed to mice . No significant increase in the intestinal antibody nor the serum IgA antibody response was detected but levels of serum IgG anti-CT antibody were significantly elevated in the group fed CT in phosphatidylcholine-based liposomes . Levels of antibody were significantly reduced in the groups fed CT in dipalmitoylphosphatidylcholine liposomes . Escherichia coli wall extract (ECWE) entrapped in certain liposome types and fed to mice elicited significantly increased serum anti-ECWE antibody responses but intestinal antibody responses were insignificantly different from the controls . These results suggest that orally administered liposomes fail to act as potent intestinal adjuvants for the entrapped antigens of bacterial origin used in this study. J Comp Pathol, 1992 Apr, 106(3), 201 - 11 Pathology of the mucous coat of trout skin during an erosive bacterial dermatitis: a technical advance in mucous coat stabilization for ultrastructural examination; Speare DJ et al.; A fixation regime which combined cryopreservation, freeze drying and vapour fixation with osmium tetroxide, was found to preserve the mucous coat of trout skin for ambient temperature scanning electron microscopy . The regime was used to study changes to the mucous coat of trout skin during a spontaneous outbreak of "columnaris" disease--a common dermatitis of commercial salmonids associated with the bacterial pathogen Cytophaga columnaris . Infected and damaged regions of skin were covered by a mucous coat which differed from that which covered adjacent unaffected areas . In unaffected areas, the mucous coat topography was smooth and relatively featureless . In contrast, the mucous coat which covered damaged areas was fissured, cratered and contained exfoliated epithelial cells . Nevertheless, this study showed that even sites of extensive dermal ulceration, in which mucous cells had been destroyed, retained a partial mucous coat . This suggests that mucus, after secretion, flows over the skin surface, rather than functioning only near the site of production . Because of the various protective functions attributed to the mucous coat, its partial presence over areas of skin damage would contribute to defence against secondary pathogens and to the prevention of excess movement of ions and water at these sites . The technical development in mucous stabilization, described in this paper, will provide a means for examining morphological changes to the mucous coat of fish skin in response to a range of stimuli in future studies. Biol Chem Hoppe Seyler, 1992 Apr, 373(4), 171 - 6 Evidence for tyrosine-linked glycosaminoglycan in a bacterial surface protein; Peters J et al.; The S-layer protein of Acetogenium kivui was subjected to proteolysis with different proteases and several high molecular mass glycosaminoglycan peptides containing glucose, galactosamine and an unidentified sugar-related component were separated by molecular sieve chromatography and reversed-phase HPLC and subjected to N-terminal sequence analysis . By methylation analysis glucose was found to be uniformly 1,6-linked, whereas galactosamine was exclusively 1,4-linked . Hydrazinolysis and subsequent amino-acid analysis as well as two-dimensional NMR spectroscopy were used to demonstrate that in these peptides carbohydrate was covalently linked to tyrosine . As all of the four Tyr-glycosylation sites were found to be preceded by valine, a new recognition sequence for glycosylation is suggested. Sci Total Environ, 1992 Apr, 114, 25 - 36 Long-range changes in oxytetracycline concentration and bacterial resistance toward oxytetracycline in a fish farm sediment after medication; Samuelsen OB et al.; Following 10 days medication with oxytetracycline, marine sediment was sampled beneath three selected cages (cages 1, 2 and 3) at a fish farm over a period of 18 mnd., in order to detect any change in the sediment oxytetracycline concentration, bacterial number and bacterial resistance towards the drug . The bulk of oxytetracycline disappeared during the first weeks, but it persisted in the sediment at lower concentrations for quite some time after the medication . Half-life (t(1/2)) of oxytetracycline in the sediment was measured as: 125, 144 and 87 days under cages 1, 2 and 3, respectively . At the end of the medication, all three sediments had greater than 100% oxytetracycline-resistant bacteria . This value dropped to 20% after 72 days and stabilised at levels of between 10 and 50% . The change in bacterial numbers, described as total and plate counts, was due to seasonal variations rather than to the medication. Trends Biochem Sci, 1992 Apr, 17(4), 150 - 4 Herbicide binding in the bacterial photosynthetic reaction center; Sinning I; The ureas and phenolics are two major classes of herbicides that act on Photosystem II (PSII) and are normally inactive in the photosynthetic reaction centers of purple bacteria . However, the triazine-resistant mutant T4 from Rhodopseudomonas (Rps.) viridis, which has the tyrosine residue at position 222 on the L subunit substituted for phenylalanine (TyrL222Phe), is sensitive to both ureas and phenolics . Since for the first time structural data on urea binding are available, T4 is a particularly interesting model for the herbicide-binding site of PSII. An Med Interna, 1992 Apr, 9(4), 175 - 7 {Bacterial pneumonia in the acquired immunodeficiency syndrome}; Hernandez-Flix S et al.; Sixty five patients with AIDS and clinical and/or radiological evidence of pulmonary infection underwent 78 bronchofibroscopies (BF) with protected brushing and bronchoalveolar washing-out . Out of the 78 BF, bacterial infection was diagnosed in 30 cases and associated opportunistic infection in 12 cases . The 18 cases of exclusively bacterial infection accounted for 23% of the total and most of them were due by H . influenzae and pneumococcus . Just in one patient, the thoracic radiography showed a localized infiltration . Given the high incidence of bacterial infections observed, along with the relevance of myxoid infections (opportunistic and pyogenic bacteria) and the low specificity of the thoracic radiography, bronchoalveolar washing-out and protected brushing in the same BF is a recommended practice. Neurology, 1992 Apr, 42(4), 739 - 48 Bacterial meningitis in children: pathophysiology and treatment; Ashwal S et al.; Recent studies of the pathophysiology of bacterial meningitis have suggested that the development of neuronal injury is related to the release of vasoactive substances or alteration of blood-brain barrier permeability . Cerebral edema, increased intracranial pressure (ICP), systemic hypotension, decreased cerebral perfusion pressure, vascular inflammation, thrombosis, and a variety of other vascular changes may result in global or regional reductions in cerebral blood flow (CBF), which contribute to this insult . Approximately one-third of infants and children with bacterial meningitis will have markedly reduced CBF, and even in those children with normal total flow, regional hypoperfusion is common . Reduced CBF is associated with cerebral edema and a poor prognosis . A poor prognosis also is associated with reduced cerebral perfusion pressure . This occurs early in the course of meningitis and is primarily due to increased ICP rather than systemic hypotension . Autoregulation is preserved, suggesting that local ischemic tissue injury is more related to factors such as regional edema formation, focal vascular pathology, or specific intrinsic flow/metabolic abnormalities than to a reduction in systemic blood pressure . In contrast with other acute CNS insults, CBF/PCO2 reactivity is well preserved in many patients with meningitis; this raises the possibility that hyperventilation may cause further ischemic injury in those patients with marginal CBF . Although it is still unclear that treatment of increased ICP will affect outcome, we propose a treatment paradigm based on the results of neuroimaging studies and ICP measurements. EMBO J, 1992 Apr, 11(4), 1251 - 9 Control of gene expression in tobacco cells using a bacterial operator-repressor system; Wilde RJ et al.; We have investigated the efficacy of using the Escherichia coli lac operator-repressor system to control plant gene expression . The lacI gene was modified to allow optimal expression in plant cells and then placed downstream of the cauliflower mosaic virus (CaMV) 35S RNA promoter . This construct was introduced into tobacco plants by leaf disc transformation . Transgenic tobacco plants synthesized significant quantities of LacI protein (up to 0.06% of total soluble protein) . We have used the E.coli beta-glucuronidase gene (gus) as the reporter gene by placing it downstream of the maize chlorophyll a/b binding protein (CAB) gene promoter . Lac operators were introduced into several positions within the CAB promoter and operator-free plasmid was used as control . Repression was assessed by comparing the transient expression from CAB-operator-gus reporter constructs in protoplasts expressing lac protein, with that in control cells not expressing the repressor . Repression varied between 10 and 90% with different operator positions . Transient assays were also performed in the presence of the inducer, isopropyl-beta-D-thiogalactoside (IPTG) . In lacI protoplasts the presence of IPTG manifested itself in a 4.2-fold relief of repression . The study was extended to show regulation of expression in stable transformants . Tobacco transformants harbouring a CAB-operator-gus reporter construct and the lacI gene were shown to have repressed GUS levels, but in the presence of IPTG, repression was relieved 15-fold . We conclude that the lac repressor can enter the plant cell nucleus, find its cognate operator sequence in the chromatin to form a repressor--operator complex and effectively block transcription of a downstream gene. Clin Exp Immunol, 1992 Apr, 88(1), 124 - 8 Effects of haemorrhage on bacterial antigen specific pulmonary plasma cell function; Robinson A et al.; Nosocomial pneumonia is frequent after haemorrhage and trauma, and often contributes to multiple organ system failure, morbidity and mortality in this setting . Although the percentages and numbers of bacterial polysaccharide antigen-specific pulmonary B cell clonal precursors are markedly decreased after haemorrhage, the effects of haemorrhage on pulmonary plasma cells actually producing antibody to these antigens are unknown . To investigate this question, the numbers of intraparenchymal pulmonary plasma cells producing antibody against the bacterial polysaccharide antigen levan (from Aerobacter levanicum) as well as bacterial antigen specific secretory IgA (sIgA) titres in the lungs were determined at various time points after 30% blood volume haemorrhage . Reduced numbers of bacterial antigen specific pulmonary plasma cells were found for more than 21 days following haemorrhage . An almost complete disappearance from the lungs of levan specific plasma cells occurred between 3 and 21 days after blood loss . Titres of bacterial antigen specific sIgA in the lungs were decreased starting at 3 days post-haemorrhage and remained significantly depressed for more than 35 days after blood loss . These results demonstrate that haemorrhage produces profound and long-lasting suppression in bacterial antigen-specific pulmonary plasma cell function . Because these effects do not occur immediately post-haemorrhage, immunization techniques able to enhance bacterial antigen specific sIgA titres at pulmonary surfaces may be able to increase resistance to nosocomial pneumonia if administered shortly after injury and blood loss. J Trop Med Hyg, 1992 Apr, 95(2), 87 - 94 Cerebrospinal fluid levels of lysozyme, IgM and C-reactive protein in the identification of bacterial meningitis; Ribeiro MA et al.; Lysozyme (LZM), immunoglobulin M (IgM) and C-reactive protein (CRP) levels were determined in cerebrospinal fluid (CSF) from patients classified on the basis of clinical and laboratory findings into three groups: bacterial meningitis (n = 33), lymphocytic meningitis (n = 21) and controls (n = 54) . IgM and CRP levels were determined by enzyme-linked immunosorbent assay (ELISA) and LZM by the lysoplate method . Discriminant analysis demonstrated that 93.94% (31/33) and 96.97% (32/33) of patients with bacterial meningitis were correctly classified on the basis of CSF determinations of IgM and LZM, respectively . However, the measurement of CRP levels in CSF correctly classified 100% of these patients (33/33), thus representing a useful additional marker for the screening of bacterial meningitis . Moreover, no more than 4% (3/75) of patients were incorrectly classified as belonging to the bacterial group on the basis of the CRP test . Thus, CRP titres less than or equal to 80 identify cases belonging to one of the non-bacterial groups, whereas titres greater than or equal to 640 classify the bacterial group, with a very low chance of misclassification . The authors recommend that CSF IgM or LZM levels be also measured for patients with CSF CRP titres of 160 and 320, for a more accurate diagnosis . The probability of these cases being of bacterial aetiology, as calculated from the combined results of these measurements, is presented. Proc Natl Acad Sci U S A, 1992 Apr 1, 89(7), 2699 - 702 16S rRNA phylogenetic analysis of the bacterial endosymbionts associated with cytoplasmic incompatibility in insects; O'Neill SL et al.; Bacterial endosymbionts of insects have long been implicated in the phenomenon of cytoplasmic incompatibility, in which certain crosses between symbiont-infected individuals lead to embryonic death or sex ratio distortion . The taxonomic position of these bacteria has, however, not been known with any certainty . Similarly, the relatedness of the bacteria infecting various insect hosts has been unclear . The inability to grow these bacteria on defined cell-free medium has been the major factor underlying these uncertainties . We circumvented this problem by selective PCR amplification and subsequent sequencing of the symbiont 16S rRNA genes directly from infected insect tissue . Maximum parsimony analysis of these sequences indicates that the symbionts belong in the alpha-subdivision of the Proteobacteria, where they are most closely related to the Rickettsia and their relatives . They are all closely related to each other and are assigned to the type species Wolbachia pipientis . Lack of congruence between the phylogeny of the symbionts and their insect hosts suggest that horizontal transfer of symbionts between insect species may occur . Comparison of the sequences for W . pipientis and for Wolbachia persica, an endosymbiont of ticks, shows that the genus Wolbachia is polyphyletic . A PCR assay based on 16S primers was designed for the detection of W . pipientis in insect tissue, and initial screening of insects indicates that cytoplasmic incompatibility may be a more general phenomenon in insects than is currently recognized. Proc Natl Acad Sci U S A, 1992 Apr 1, 89(7), 2600 - 4 Expression of a bacterial mtlD gene in transgenic tobacco leads to production and accumulation of mannitol; Tarczynski MC et al.; A bacterial gene encoding mannitol-1-phosphate dehydrogenase, mtlD, was engineered for expression in higher plants . Gene constructions were stably incorporated into tobacco plants . The mtlD gene was expressed and translated into a functional enzyme in tobacco, resulting in the synthesis and accumulation of mannitol, which was identified by NMR and mass spectroscopy . Mannitol concentrations exceeded 6 mumol/g (fresh weight) in the leaves and in the roots of some transformants, whereas this sugar alcohol was not detected in these organs of wild-type tobacco plants or of untransformed tobacco plants that underwent the same regeneration scheme . These experiments demonstrate that branch-points in plant carbohydrate metabolism can be generated by which novel gene products can utilize endogenous substrates to divert metabolic energy into novel compounds . Additionally, the system described here allows for physiological studies in which the responses of wild-type and transgenic tobacco to various environmental stimuli can be compared directly . Such studies will facilitate our understanding of the roles of sugar alcohols (e.g., in stress tolerance) in higher plants. Pediatrics, 1992 Apr, 89(4 Pt 1), 640 - 2 No increased risk for invasive bacterial infection found following diphtheria-tetanus-pertussis immunization; Griffin MR et al.; During the acellular pertussis vaccine trial in Sweden, 4 children who were randomly assigned to receive the vaccine died of suspected or confirmed bacterial infections compared to 1 expected . There were no deaths in the placebo arm . This raised concern about the role of pertussis immunization in the development of serious infections . Through linking computerized immunization records with an active surveillance system for serious bacterial infections in children, the authors studied a cohort of 64,591 children immunized through Tennessee county health clinics who had a total of 158 episodes of invasive bacterial infections after a diphtheria and tetanus toxoids and pertussis (DTP) immunization . There were 8 invasive bacterial infections that occurred within the first 7 days following DTP immunization, yielding an age-adjusted relative risk (95% confidence interval) of 1.0 (0.5 to 2.0), compared to the interval 29 or more days following immunization . There were 7 and 20 infections in the 8- through 14- and 15- through 28-day intervals following DTP immunization, giving relative risks of 0.8 (0.4 to 1.7) and 1.2 (0.7 to 1.9), respectively . These data provide reassurance that the use of DTP vaccine is not followed by a large increased risk of serious bacterial infections. Hybridoma, 1992 Apr, 11(2), 225 - 37 Monoclonal antibodies against bacterial glucosamine 6-phosphate synthase: production and use for structural studies; Cochet O et al.; Fifteen mouse x rat hybridoma cell lines producing rat monoclonal antibodies (MAbs) directed to Escherichia coli Glucosamine 6-P Synthase (GlmS) were established and characterized . Most of them (13/15) are IgG2a while 2 were typed as IgG1 . Their Kaff ranged from 1.5 x 10(6) to 9.6 x 10(8) M-1 as determined by Beatty et al . (1) . The epitopes recognized by these MAbs were assigned to one of the two catalytical domains of the enzyme (CT1 and CT2) as demonstrated both by ELISA and Western-blotting using purified GlmS proteolytic fragments . The binding of the MAbs on either the native or denatured forms of GlmS, CT1 and CT2 was further analyzed by competitive immunoassay and most of the MAbs were found to bind preferentially to the denatured proteins . The study of the antigenic topography of GlmS by competitive radioimmunoassay demonstrated the existence of at least 10 independent epitopes on GlmS, divided into three groups . The first one (3/15) includes MAbs whose binding was not inhibited by any of the other MAbs . The second group (9/15) is comprised of MAbs that exhibit reciprocal binding inhibitory activity while the third group includes MAbs (3/15) presenting asymmetric inhibitory activity . Finally, since most of the isolated antibodies (10/15) bind to the 27 kDa amino-terminal glutamine binding domain (CT2), the capacity of these MAb to interfere with the associated glutaminase activity was analyzed. J Clin Microbiol, 1992 Apr, 30(4), 1039 - 41 Chemiluminescent universal probe for bacterial ribotyping; Gustaferro CA et al.; We describe a novel procedure for direct covalent coupling of horseradish peroxidase to rRNA and ribotyping by using enhanced chemiluminescence . Compared with their 32P-end-labeled counterparts, chemiluminescent rRNA probes are stable and easy to synthesize and provide results as good as or superior to those obtained with isotopic labeling . Direct chemiluminescent labeling of Escherichia coli rRNA produces a sensitive, universal probe suitable for clinical laboratory use in the investigation of nosocomial outbreaks. Int J Radiat Biol, 1992 Apr, 61(4), 511 - 8 Radioprotection of mice by the bacterial extract Broncho-Vaxom: haemopoietic stem cells and survival enhancement; Fedorocko P et al.; Pretreatment of mice with 50-1000 micrograms of the bacterial extract Broncho-Vaxom (BV, free of endotoxin) before sublethal irradiation induced an increase in the number of endogenous haemopoietic stem cells (E-CFU) . The degree of radioprotection was dependent on both the time of administration and the dose of BV . An optimal E-CFU survival was observed when 500 micrograms of BV was administered i.p . 24 h before irradiation . BV did not affect the day 9 CFU-S survival in the bone marrow directly after irradiation . However, 5, 9 and 12 days after irradiation, the number of day 9 CFU-S was almost 2-fold higher in the bone marrow of BV injected mice . Pretreatment with BV protected C57B1/6 mice in a dose-dependent manner from the lethal effect of ionizing radiation . A single dose (50, 100, 250, or 500 micrograms) of bacterial lysate injected i.p . 24 h before 9.5 Gy gamma-rays (LD100/21) protected 16%, 25%, 80%, and 94% of C57B1/6 mice, respectively . The dose reduction factor in the case when the BV (500 micrograms per mouse) was administered at that time was 1.18 (95% CL 1.12, 1.25). Pneumoftiziologia, 1992 Apr-Sep, 41(2-3), 135 - 9 {The characteristics of bacterial respiratory infections in patients with humoral immunodeficiencies}; Dobre V; The paper deals with 8 cases of different forms of bacterial respiratory infections (bronchiectasis, pleuropneumonia, tuberculosis), in which humoral immune deficits were noted: 2 dysgammaglobulinemia with IgA absence, 3 hypogammaglobulinemia with IgG deficit as well as IgA and IgM absence, 2 hypogammaglobulinemia with IgG and IgM deficit and IgA absence, and 1 hypogammaglobulinemia with IgA absence and IgM deficit . Stress is laid on the indications given by the antibody titer research, the pleural exudate hypoproteinemia being also added to those already known . A correct chemotherapy associated with the substitution treatment proved to by efficient. J Endod, 1992 Apr, 18(4), 166 - 71 Dentin as inhibitor of bacterial toxicity on pulpal cells in vitro; Pissiotis E et al.; Many studies have shown that the major cause of pulpal disease is the presence of bacteria or their by-products in the dentinal tubules . The purpose of this investigation was to develop an in vitro model, simulating the pulp chamber, that would permit the study of the transport of bacterial by-products through dentin and their effect on pulpal cells . Human pulpal cells were cultured in a modified Sykes-Moore chamber and exposed through dentin to sonicated extracts of Porphyromonas gingivalis ATCC 33277 . The cell response was evaluated with the thymidine incorporation method . The results were compared with the cell response obtained after direct exposure to the same irritant . It was found that dentin significantly restricts the diffusion of bacterial proteins in a 24-h experimental period . The time needed for the first bacterial protein molecule to cross the dentin barrier was 6 h . The "diffusion velocity" of the bacterial proteins was 0.023 microns/s . The proposed model has further applications in biocompatibility and microleakage research. Oral Microbiol Immunol, 1992 Apr, 7(2), 111 - 2 Bacterial synergy of Treponema denticola and Porphyromonas gingivalis in a multinational population; Simonson LG et al.; Treponema denticola and Porphyromonas gingivalis have been associated with human adult severe periodontitis . In this study, we quantified these putative pathogens in subgingival plaque samples collected from 74 Fijians, 74 Colombians and 73 U.S . Americans stationed at the Multinational Force and Observers encampment in the Sinai Desert, Egypt . A contingency table of T . denticola and P . gingivalis frequency revealed a highly significant synergistic relationship . We discovered that the occurrence of T . denticola apparently requires the presence of P . gingivalis . This represents the first observation of a synergistic relationship between these putative oral pathogens associated with adult severe periodontal disease. Virus Res, 1992 Apr, 23(1-2), 27 - 38 Bacterial expression of the capsid antigen domain and identification of native gag proteins in spumavirus-infected cells; Bartholoma A et al.; A bacterial expression plasmid containing the central part of the gag gene of the human spumaretrovirus (HSRV) was constructed and expressed in E . coli . The expected protein product consisting of the complete region of the HSRV capsid antigen and part of the matrix protein was expressed in relatively large amounts . Polyclonal antisera raised against this recombinant protein were used to identify authentic gag precursors of 78 and 74 kDa and processed gag proteins of 60, 58, and 33 kDa in HSRV-infected human embryonal fibroblast cells by radioimmunoprecipitation . The recombinant antigen will be useful for the detection of antibodies against HSRV gag proteins in human sera. Infect Immun, 1992 Apr, 60(4), 1455 - 64 Porphyromonas gingivalis virulence in mice: induction of immunity to bacterial components; Kesavalu L et al.; Selected cell envelope components of Porphyromonas gingivalis were tested in a BALB/c mouse model in an attempt to elucidate further the outer membrane components of this putative oral pathogen that might be considered as virulence factors in host tissue destruction . Lipopolysaccharide (LPS), outer membrane, and outer membrane vesicles of P . gingivalis W50, ATCC 53977, and ATCC 33277 were selected to examine an immunological approach for interference with progressing tissue destruction . Mice were actively immunized with heat-killed (H-K) or Formalin-killed (F-K) whole cells or with the outer membrane fraction, LPS, or outer membrane vesicles of the invasive strain P . gingivalis W50 . The induction of invasive spreading lesions with tissue destruction and lethality were compared among different immunization groups in normal, dexamethasone-treated (dexamethasone alters neutrophil function at the inflammatory site), and galactosamine-sensitized (galactosamine sensitization increases endotoxin sensitivity) mice after challenge infection with the homologous strain (W50) and heterologous strains (ATCC 53977 and ATCC 33277) . Enzyme-linked immunosorbent assay analyses revealed significantly elevated immunoglobulin G and M antibody responses after immunization with H-K or F-K cells or the outer membrane fraction compared with those of nonimmunized mice . The killed whole-cell vaccines provided significantly greater protection against challenge infection in normal mice (decreased lesion size and death) than did either the outer membrane fraction or LPS immunization . The lesion development observed in dexamethasone-pretreated mice was significantly enhanced compared with that of normal mice after challenge with P . gingivalis . Immunization with P . gingivalis W50 provided less protection against heterologous challenge infection with P . gingivalis ATCC 53977; however, some species-specific antigens were recognized and induced protective immunity . Only viable P . gingivalis induced a spreading lesion in normal, dexamethasone-treated, or galactosamine-sensitized mice; F-K or H-K bacteria did not induce lesions . The F-K and outer membrane vesicle immunization offered greater protection from lesion induction than did the H-K immunogen after challenge infection simultaneous with galactosamine sensitization . The H-K cell challenge with galactosamine sensitization produced 100% mortality without lesion induction, suggesting that LPS or LPS-associated outer membrane molecules were functioning like endotoxin . Likewise, P . gingivalis W50 LPS (1 micrograms per animal) administered intravenously produced 80% mortality in galactosamine-sensitized mice . In contrast to the effects of immunization on lesion development, immunization with H-K or F-K cells or LPS provided no protection against intravenous challenge with LPS; 100% of the mice died from acute endotoxin toxicity.(ABSTRACT TRUNCATED AT 400 WORDS) J Immunol, 1992 Apr 1, 148(7), 2186 - 93 Bacterial lipopolysaccharide up-regulates platelet-activating factor-stimulated Ca2+ mobilization and eicosanoid release in human Mono Mac 6 cells; Aepfelbacher M et al.; When human monocytic Mono Mac 6 cells were treated with bacterial LPS (10 ng/ml, 72 h), they showed an increase in phagocytic activity, superoxide anion production, and expression of monocyte/macrophage-associated cell surface Ag . In these more mature (LPS-treated) cells but not in untreated cells, platelet-activating factor (PAF) (100 nM) produced a three- to fourfold increase in cytosolic free Ca2+ concentration . The cytosolic free Ca2+ concentration increase was inhibited by the PAF receptor antagonist L-659,989 (10 microM) and by EGTA (2 mM), indicating receptor-dependent Ca2+ influx . Furthermore, L-659,989 (10 microM), as well as PAF (1 microM), inhibited specific {3H}PAF binding in LPS-treated but not in untreated cells . Consistent with these results, PAF (100 nM) stimulated release of arachidonic acid and thromboxane B2 only in LPS-treated cells, and this could be inhibited by L-659,989 (10 microM) and EGTA (2 mM) . Our data indicate that LPS up-regulates PAF-induced Ca2+ influx, resulting in arachidonic acid and eicosanoid release in Mono Mac 6 cells. Am J Med, 1992 Apr, 92(4), 391 - 5 Increased risk of bacterial endocarditis in inflammatory bowel disease; Kreuzpaintner G et al.; PURPOSE: The purpose of this retrospective as well as prospective case-control study was to analyze a possible overrepresentation of inflammatory bowel diseases among patients with native valve endocarditis as well as the factors that predispose patients with inflammatory bowel disease to infective endocarditis . PATIENTS AND METHODS: Among 213 consecutive patients treated for proven native valve endocarditis, six (2.8%) had inflammatory bowel diseases (three with ulcerative colitis and three with Crohn's disease) . Three patients with inflammatory bowel disease were from the retrospective group, and three were from the prospective group . The prevalence of inflammatory bowel diseases has been determined to be 0.0641% in the Dusseldorf area . RESULTS: On the basis of these data, a 44-fold overrepresentation of inflammatory bowel diseases among the 213 patients with endocarditis was calculated with a statistical significance of p much less than 0.001 . CONCLUSIONS: Inflammatory bowel disease may be considered an independent risk factor for bacterial endocarditis . Reasons may be more frequent bacteremias as a result of the higher incidence of diagnostic and therapeutic interventions, as well as increased permeability of the damaged mucosa for bacteria and the therapeutic immunosuppression in patients with active inflammatory bowel disease . Prophylaxis for bacterial endocarditis should be carefully considered before expected bacteremias in patients with highly active inflammatory bowel disease even in the absence of cardiac factors predisposing to bacterial endocarditis. J Biol Chem, 1992 Mar 15, 267(8), 5614 - 20 Active site topologies of bacterial cytochromes P450101 (P450cam), P450108 (P450terp), and P450102 (P450BM-3) . In situ rearrangement of their phenyl-iron complexes; Tuck SF et al.; The reactions of cytochromes P450101 (P450cam), P450108 (P450terp), and P450102 (P450BM-3) with phenyldiazene result in the formation of phenyl-iron complexes with absorption maxima at 474-478 nm . Treatment of the cytochrome P450 complexes with K3Fe(CN)6 decreases the 474-478 nm absorbance and shifts the phenyl group from the iron to the porphyrin nitrogens . Acidification and extraction of the prosthetic group from each of the ferricyanide-treated enzymes yields a different mixture of the four possible N-phenylprotoporphyrin IX regioisomers . The ratios of the regioisomers with the phenyl ring on pyrrole rings B, A, C, and D (in order of elution from the high performance liquid chromatography column) are, respectively: cytochrome P450cam, 0:0:1:4; P450terp, 0:0:0:1; and P450BM-3, 2:10:2:1 . The isomer ratio for recombinant cytochrome P450BM-3 without the cytochrome P450 reductase domain (2:9:2:1) shows that the reductase domain does not detectably perturb the active site topology of cytochrome P450BM-3 . Potassium ions modulate the intensity of the spectrum of the phenyl-iron complex of cytochrome P450cam, but do not alter the N-phenyl isomer ratio . Computer graphics analysis of the crystal structure of the cytochrome P450cam phenyl-iron complex indicates that the active site of cytochrome P450cam is open above pyrrole ring D and, to a small extent, pyrrole ring C, in complete agreement with the observed N-phenylprotoporphyrin IX regioisomer pattern . The regioisomer ratios indicate that the active site of cytochrome P450terp is only open above pyrrole ring D, whereas that of cytochrome P450BM-3 is open to some extent above all the pyrrole rings but particularly above pyrrole ring A . The bacterial enzymes thus have topologies distinct from each other and from those of the mammalian enzymes so far investigated, which have active sites that are open to a comparable extent above pyrrole rings A and D. Science, 1992 Mar 13, 255(5050), 1437 - 40 Inhibition of development of Kaposi's sarcoma-related lesions by a bacterial cell wall complex; Nakamura S et al.; In vitro and in vivo model systems for the study of human immunodeficiency virus (HIV)-associated Kaposi's sarcoma (KS) were used to evaluate compounds for their potential as therapeutic agents . A sulfated polysaccharide-peptidoglycan compound (SP-PG) produced by bacteria controlled the in vitro growth of acquired immunodeficiency syndrome (AIDS)-associated, KS-derived spindle-shaped cells (AIDS-KS cells) at noncytotoxic concentrations . Angiogenesis induced by AIDS-KS cells in the chicken chorioallantoic membrane assay was blocked by SP-PG, which also inhibited the vascular hyperpermeability response and the angiogenesis associated with the induction of KS-like lesions that develop after subcutaneous inoculation of AIDS-KS cells into nude mice . Suramin, pentosan polysulfate, and interferon alpha, which are currently in use for therapy of KS, were either less effective than SP-PG or much more cytotoxic, or both. Biochim Biophys Acta, 1992 Mar 12, 1119(3), 322 - 6 Evidence that the methylesterase of bacterial chemotaxis may be a serine hydrolase; Krueger JK et al.; CheB, the methylesterase of chemotactic bacteria, catalyzes the hydrolysis of glutamyl-methyl esters in bacterial chemoreceptor proteins . The two cysteines predicted by the amino acid sequence of CheB were replaced by alanine residues . The resulting mutants, Cys207-Ala, Cys309-Ala and a double cysteine mutant Cys207-Ala/Cys309-Ala, retained methylesterase activity, indicating that sulfhydryls are not crucial for CheB mediated catalysis . A homology search revealed a conserved serine active-site region between residues 162 and 166 which is homologous to the active-site region of acetylcholine esterases, suggesting that Ser164 of CheB is the active-site nucleophile . Oligonucleotide-directed mutagenesis was used to change the serine to a cysteine . This Ser164-Cys mutant had less than 2% of the wild-type activity . Unlike the serine proteinases which utilize a 'catalytic triad' mechanism, CheB does not have the conserved histidine and aspartic acid residues located in positions N-terminal to the active-site serine . In addition, CheB is not labeled with di-isopropylfluorophosphate, a potent inhibitor of other serine hydrolases . A novel mechanism is proposed for CheB involving substrate-assisted catalysis to account for these apparent anomalies. Genetics, 1992 Mar, 130(3), 677 - 83 Biased gene conversion, copy number, and apparent mutation rate differences within chloroplast and bacterial genomes; Birky CW Jr et al.; We investigate the possibility that differences between synonymous substitution rates of organelle and bacterial genes differing only in copy number may be due to conversion bias . We find that the rather large observed difference in the synonymous rates between genes in the single copy and inverted-repeat regions of chloroplasts can be accounted for by a very small bias against new mutants . More generally, differences in the within-organelle fixation probability result in different apparent mutation rates as measured by the expected rate of appearance of cells homoplasmic for new mutants . Thus, differences in intracellular population parameters rather than molecular mechanisms can account for some variation in the apparent mutation rates of organelle genes, and possibly in other systems with variable numbers of gene copies . On the other hand, our analysis suggests that conversion bias is not a likely explanation for relatively low mutation rates observed near the replication origin of bacterial chromosomes. J Biol Chem, 1992 Mar 5, 267(7), 4664 - 71 Molecular cloning, chromosomal localization, and bacterial expression of a murine macrophage metalloelastase; Shapiro SD et al.; Murine macrophages have previously been shown to secrete a zinc-dependent proteinase that can degrade elastin . In this report, we identify murine macrophage elastase (MME) cDNA and show that it is a distinct member of the metalloproteinase gene family . Small amounts of MME were purified to homogeneity, and N-terminal amino acid sequence was obtained . This sequence was used to obtain a partial cDNA clone by the polymerase chain reaction; a cDNA library derived from a mouse macrophage-like cell line (P388D1) was screened with this probe . A full-length MME cDNA spanning approximately 1.8 kilobases contained an open reading frame of 1386 base pairs; the predicted molecular mass of the MME proenzyme is 53 kDa . The gene encoding MME is represented only once in the mouse genome and is located on chromosome 9 . Despite a size that is similar to other metalloproteinases, MME is distinct, sharing only 33-48% amino acid homology with other metalloproteinases . In contrast to other metalloenzymes, MME appears to be rapidly processed to an active truncated form (N-terminal and C-terminal cleavage) . We expressed recombinant MME in Escherichia coli and demonstrated that it has significant elastolytic activity that is specifically inhibited by the tissue inhibitor of metalloproteinases . MME is therefore a true metalloproteinase that may be involved in tissue injury and remodeling. Exp Cell Res, 1992 Mar, 199(1), 169 - 73 Mitogenic effects of bacterial neuroaminidase and lactosylceramide on human cultured fibroblasts; Ogura K et al.; Exogenously added bacterial neuraminidase and lactosylceramide both stimulated the growth of cultured human skin fibroblasts . Neuraminidase (100 units/ml) increased DNA synthesis 1.9-fold and cell density 1.4-fold after 24 and 48 h, respectively, in culture . Treated fibroblasts contained less ganglioside NeuAc alpha 2-3Gal beta 1-4GlcCer (GM3), presumably due to neuraminidase-catalyzed hydrolysis to lactosylceramide . Addition of lactosylceramide (100 microM) to the fibroblast culture medium also increased DNA synthesis threefold within 24 h and cell density twofold after 48 h . These findings are compatible with a mechanism by which the proliferation of human fibroblasts is regulated by the relative levels of GM3 and lactosylceramide in the plasma membrane. J Chir (Paris), 1992 Mar, 129(3), 155 - 9 {Bacterial endocarditis complicated by popliteal and mesenteric aneurysms}; Christides C et al.; The authors report about one case of bacterial endocarditis complicated by fungal aneurysms in a superior mesenteric and a popliteal site . While the diagnosis was easy for the popliteal aneurysm, it was not so for the mesenteric aneurysm . Arteriography must have wide indications . The treatment of such aneurysms must always be medical, but surgical as well . The surgical tactics must be carefully discussed, and the restoration of vascular continuity with autologous venous material through an extra-anatomic course should be preferred. Clin Ther, 1992 Mar-Apr, 14(2), 306 - 13 A noncomparative study of cefprozil at two dose levels in the treatment of acute uncomplicated bacterial sinusitis; van den Wijngaart W et al.; The efficacy and safety of cefprozil at two dose levels were evaluated in 110 patients with acute uncomplicated bacterial sinusitis in an uncontrolled, noncomparative, Phase II trial . Ninety patients received 250 mg of cefprozil (low-dose group) and 20 patients received 500 mg of cefprozil (high-dose group) every 12 hours for ten days . Evaluable patients had symptoms consistent with acute sinusitis, pathogens isolated at pretreatment susceptible to cefprozil, and a radiograph positive for sinusitis within 48 hours before treatment . A satisfactory clinical response was achieved in 34 of 39 evaluable patients (87%) in the low-dose group and in all 16 evaluable patients (100%) in the high-dose group . Pathogens were eradicated in 35 of 39 patients (90%) in the low-dose group and in 15 of 16 patients (94%) in the high-dose group . A total of 140 of 155 pathogens (90%) isolated pretreatment were susceptible to cefprozil . Six patients (7%) in the low-dose group and one patient (5%) in the high-dose group reported at least one adverse clinical event. Am J Perinatol, 1992 Mar, 9(2), 69 - 74 The effects of pH and osmolality on bacterial growth in amniotic fluid in a laboratory model; Silver HM et al.; In studying the effects of amniotic fluid on bacterial growth in a laboratory model, we noted that the pH of the fluid appeared to exert an independent effect . This study was designed to test the ability to control the growth of Escherichia coli in amniotic fluid simply by controlling two important growth conditions, pH and osmolality . The effects of pH and osmolality on growth of E . coli were systematically studied in a standard media and in amniotic fluid . Optimal ranges in standard media were pH of 5.6 to 6.6 and osmolality of 150 to 215 mOsm . When the results of growth at 24 hours were corrected for pH by analysis of covariance, the presence of amniotic fluid or phosphate had no effect . We found pH to be the only variable predictive of bacterial growth in amniotic fluid in this laboratory model. Pediatr Neurol, 1992 Mar-Apr, 8(2), 142 - 4 Uncal herniation secondary to bacterial meningitis in a newborn; Feske SK et al.; A newborn with bilateral uncal herniation secondary to acute bacterial meningitis is reported . The findings of previous neuropathologic studies of neonatal bacterial meningitis are reviewed and the factors most likely responsible for the relative rarity of herniation in this disease in newborns are discussed. Gut, 1992 Mar, 33(3), 307 - 11 Activation of the classical complement pathway in spontaneous bacterial peritonitis; Bird G et al.; To investigate the possibility that low complement concentrations in the plasma and ascites of patients with severe liver disease could be secondary to complement consumption, complement activation was studied in 32 patients with severe liver disease, 11 of whom had spontaneous bacterial peritonitis (SBP) . In patients with SBP, plasma C3 and C4 were significantly lower than in uninfected patients (mean values 0.74 v 1.13 g/l, p less than 0.01 and 0.20 v 0.28 g/l, p less than 0.05 respectively) . Plasma complement activation via the classical pathway, as shown by C4d/C4, was significantly increased in patients with SBP compared with uninfected patients (37.3 v 22.2, p less than 0.01) as was C3d/C3 (14.0 v 8.11, p less than 0.01), but there was no significant difference in Ba/B between SBP and uninfected patients . Ascitic C3 concentrations were higher in patients without SBP than in infected patients (0.37 v 0.08 g/l, p less than 0.05), as were factor B values (0.11 v 0.03 g/l, p less than 0.05) . There was no significant difference in ascitic C4 concentrations in patients with SBP compared with uninfected patients (0.03 v 0.07 g/l) . Although consumption of C3, as shown by C3d/C3 in ascites, was increased in infected patients compared with uninfected patients (79.1 v 36.1, p less than 0.05), there was no difference in ascitic complement activation between the groups for either the classical or alternative pathways . In SBP, decreased plasma C3 and C4 are primarily caused by increased activation of the classical pathway and not impaired hepatic synthesis . Activation and consumption of C3 is one factor causing the low ascitic C3 concentrations observed in SBP. Int J Pediatr Otorhinolaryngol, 1992 Mar, 23(2), 117 - 24 Bacterial quantification--a necessary complement for the comprehension of middle ear inflammations; Raisanen S et al.; Quantification of bacteria in various types of middle ear effusion (MEE) obtained during current acute otitis media (AOM), otitis media with effusion (OME) and chronic suppurative otitis media (COM) was performed . The bacteria were stained with acridine orange and their number per ml effusion evaluated under the fluorescence microscope according to a method described in detail elsewhere . During AOM, 53% of the MEE samples were culture-positive and contained 10(6)-10(8) bacteria per ml (median value 10(7) per ml) . During OME, serous effusion and 78% of the mucoid effusions contained no bacteria whatsoever, whereas the remaining mucoid effusions contained 10(4)-5 x 10(5) bacteria per ml (median value 10(4) per ml) . Mucopurulent effusions contained 6 x 10(5)-10(8) bacteria per ml (median value 5 x 10(6) per ml) . During COM, purulent MEE had 6 x 10(6)-10(9) bacteria per ml (median value 10(8) per ml) . Quantification of bacteria involved in middle ear diseases provides further information about the etiopathogenesis and appropriate management of various pathological conditions of the middle ear. Genomics, 1992 Mar, 12(3), 534 - 41 The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries; Bentley DR et al.; An automated gridding procedure for the inoculation of yeast and bacterial clones in high-density arrays has been developed . A 96-pin inoculating tool compatible with the standard microtiter plate format and an eight-position tablet have been designed to fit the Biomek 1000 programmable robotic workstation (Beckman Instruments) . The system is used to inoculate six copies of 80 x 120-mm filters representing a total of approximately 20,000 individual clones in approximately 3 h . High-density arrays of yeast artificial chromosome (YAC) and cosmid clones have been used for rapid large-scale hybridization screens of ordered libraries . In addition, an improved PCR library screening strategy has been developed using strips cut from the high-density arrays to prepare row and column DNA pools for PCR analysis . This strategy eliminates the final hybridization step and allows identification of a single clone by PCR in 2 days . The development of automated gridding technology will have a significant impact on the establishment of fully versatile screening of ordered library resources for genomic studies. Am Rev Respir Dis, 1992 Mar, 145(3), 626 - 31 Increased adherence of monocytes to fibronectin in bronchiectasis . Regulatory effects of bacterial lipopolysaccharide and role of CD11/CD18 integrins; Owen CA et al.; Regulated adherence of monocytes to extracellular matrix is a prerequisite for accumulation of mononuclear phagocytes during pulmonary infection and inflammation . We have obtained monocytes from patients with an inflammatory lung disease (bronchiectasis) and from control subjects and have compared their adherence to fibronectin . Spontaneous adherence of monocytes from the control subjects was 20 +/- 2%, whereas that of patients' cells was markedly higher and correlated with the severity of airway inflammation: 65 +/- 5% and 40 +/- 8% in patients with purulent and mucoid sputum, respectively . Endotoxin and cytokines from areas of airway disease are likely to be responsible for the observed monocyte activation, since: (1) endotoxin was detectable in all of the patients but in none of the control subjects; (2) LPS produced a dose-related increase in adherence of normal monocytes in vitro (maximal 65 +/- 2% adherence at 1 microgram/ml of LPS); (3) recombinant cytokines and LPS produced additive effects on monocyte adherence in vitro . The adherence of the patients' monocytes to fibronectin was substantially mediated by CD11/CD18 integrins, via both RGD-dependent and RGD-independent mechanisms . These data indicate that signals arising from foci of infection and inflammation can influence the adherence of monocytes, and they are likely to be determinants of the accumulation of mononuclear phagocytes in the lungs of patients with bronchiectasis. Lab Invest, 1992 Mar, 66(3), 347 - 61 Neutrophil depletion protects against liver injury from bacterial endotoxin; Hewett JA et al.; Neutrophil (PMN) infiltration is an early occurrence in the liver after exposure to hepatotoxic doses of endotoxin lipopolysaccharide (LPS) . The purpose of this study was to test the hypothesis that PMNs contribute to the pathogenesis of LPS hepatotoxicity . The immunoglobulin fraction from serum of rabbits immunized with rat PMNs (anti-PMN Ig) was administered intravenously to rats 18 and 6 hours before exposure to an hepatotoxic dose of LPS (Escherichia coli 0128:B12) . This protocol caused a greater than 95% reduction in circulating PMNs, which was maintained for the duration of the study . The immunoglobulin fraction from nonimmunized rabbits was used as a control (control Ig) . Rats pretreated with control Ig exhibited a marked increase in the number of PMNs in the liver 1.5 hours after LPS exposure . This increase in hepatic PMNs was significantly reduced by pretreatment with anti-PMN Ig . Marked elevations in both alanine and aspartate aminotransferase activities (1086 +/- 311 and 880 +/- 183 SF units/ml, respectively) were observed in plasma from control Ig-treated rats 6 hours after intravenous administration of LPS (3.0 mg/kg) . The response to LPS was greatly attenuated in animals receiving anti-PMN Ig (145 +/- 111 and 224 +/- 49 SF units/ml alanine and aspartate aminotransferase activities, respectively) . Pretreatment of rats with immunoglobulins to rat lymphocytes reduced numbers of circulating lymphocytes but did not afford protection against the hepatotoxic effects of LPS . These results suggest that PMNs contribute to the pathogenesis of LPS hepatotoxicity. Circ Shock, 1992 Mar, 36(3), 208 - 16 Induction of early-phase tolerance to endotoxin-induced mucosal injury, xanthine oxidase activation, and bacterial translocation by pretreatment with endotoxin; Deitch EA et al.; The goal of this study was to determine whether tolerance would develop to endotoxin-induced mucosal injury, xanthine oxidase activation, and bacterial translocation . To accomplish this goal, four groups of mice were studied: 1) mice receiving ip injections of saline 96 and 24 hr prior to sacrifice, 2) mice receiving ip injections of saline 96 and endotoxin (0.1 mg) 24 hr prior to sacrifice, 3) mice receiving ip injections of endotoxin 96 and 24 hr prior to sacrifice, and 4) mice receiving ip injections of endotoxin 96 hr and saline 24 hr prior to sacrifice . In contrast to the saline control animals or mice sacrificed 96 hr after a single dose of endotoxin, mice sacrificed 24 hr after receiving a single dose of endotoxin had evidence of mucosal injury, elevated levels of ileal xanthine oxidase activity, and an 81% incidence of bacterial translocation . Mice sacrificed 24 hr after a second dose of endotoxin were largely protected against the toxic effects of endotoxin . Thus tolerance to endotoxin-induced bacterial translocation does develop and is associated with tolerance to endotoxin-induced ileal mucosal injury and xanthine oxidase activation. Indian J Med Res, 1992 Mar, 95, 88 - 92 Variations in isoenzymes of cloned & uncloned axenic Entamoeba histolytica without bacterial association; Romana S et al.; Five clones of axenic E . histolytica (HMI) grown as discrete colonies in semisolid agar medium were adapted in liquid medium and labelled as HMI-C121, HMI-C131, HMI-C143, HMI-C144 and HMI-C145 . Isoenzymes of these 5 clones of E . histolytica (HMI) were investigated in starch gel electrophoresis . There were no differences in the electromobility of maleate NADP oxidoreductase and glucosephosphoisomerase amongst the five clones and uncloned cultures of axenic E . histolytica . The relative electromobility (rf) of a single phosphoglucomutase (PGM) band of uncloned Mexican E . histolytica (HMI) and Indian axenic E . histolytica (KCG: 0986: 11) cultures and cloned E . histolytica HMI-C121, HMI-C145 was 0.087 while a single PGM band of uncloned E . histolytica (NIH: 200) and cloned E . histolytica HMI-C131, HMI-C143 and HMI-C144 cultures had rf of 0.075 . Isoenzyme characterization of four cloned HMI-C121, HMI-C131, HMI-C143, HMI-C144 cultures of axenic E . histolytica (HMI) revealed existence of three bands of hexokinase (HK) . The additional third band of HK was located close to the place of application of lysate and had rf ranging from 0.11-0.14 . The data indicated that parent axenic E . histolytica (HMI) consisted of several populations and each population expressed different isoenzyme pattern without an association of amoebic cultures with any bacterial species. Lik Sprava, 1992 Mar, (3), 50 - 3 {Mineral metabolism in bacterial meningoencephalitis}; Iarosh OO; The authors investigated the concentration of some metals in the cerebrospinal fluid in 70 patients with bacterial meningoencephalitis in the dynamics of the inflammatory process and in different regions of the brain in 10 autopsy cases . There was a correlation between potassium, sodium, manganese, magnesium, copper and zinc in the brain and quantity of water, as well as their concentration in cerebrospinal fluid . It is shown that determination of some metals in the cerebrospinal fluid allows to diagnose edema swelling of the brain and to predict the disease sequels. Hua Xi Yi Ke Da Xue Xue Bao, 1992 Mar, 23(1), 91 - 3 {Plasma fibronectin determination in acute bacterial pneumonia}; Liu L et al.; Plasma fibronectin (PFn) level was measured with immunoelectrophoresis in 40 healthy adults and 30 patients with acute bacterial pneumonia . The results showed that PFn was considerably lowered in acute bacterial pneumonia (214.49 +/- 77.84 micrograms/ml) when compared with that of healthy controls (292.48 +/- 43.11 micrograms/ml), (P less than 0.001) . In the pneumonia group (10 severe cases) the level of PFn was significantly lowered than that in the moderate or mild cases (127.45 +/- 31.03 micrograms/ml vs . 255.11 +/- 54.16 micrograms/ml), (P less than 0.001) . In 11 cases of recovering pneumonia, PFn was significantly higher than that in exacerbation period (213.13 +/- 41.32 micrograms/ml vs . 154.52 +/- 51.27 micrograms/ml) (P less than 0.001) . We think that PFn level is helpful to evaluate the clinical course and prognosis of acute bacterial pneumonia. Vet Pathol, 1992 Mar, 29(2), 169 - 74 Immune complex-mediated glomerulonephritis associated with bacterial kidney disease in the rainbow trout (Oncorhynchus mykiss); Sami S et al.; Rainbow trout (Oncorhynchus mykiss) developed a post-infectious chronic membranous glomerulonephritis 15 months after they had been experimentally infected with Renibacterium salmoninarum . Histologically, peritubular and periglomerular fibrosis, hypercellular glomeruli with occluded Bowman's space, and partial or complete adhesion to Bowman's capsule were constant features . Electron microscopy revealed thickened glomerular basement membranes with spikes accompanied by finely granular electron-dense deposits at the epithelial side and dense material in the mesangial matrix . Indirect immunofluorescence indicated linear immunoglobulin deposits along the glomerular basement membrane . The presence of R . salmoninarum was demonstrated by culture and by indirect immunofluorescence . Low serum hemagglutination-inhibiting antibody titers were demonstrated. Klin Monatsbl Augenheilkd, 1992 Mar, 200(3), 178 - 81 {Incidence of postoperative bacterial infections after planned intraocular interventions}; Rummelt V et al.; Between August 1982 and August 1984 3059 intraocular operations were performed with topical prophylactic antibiotics . Results of conjunctival cultures did not influence the surgical schedule . 8179 intraocular operations were performed between September 1984 and August 1988 . An intraocular operation was postponed until conjunctival cultures were negative using topical antibiotics administered at hourly intervals . The rate of postoperative intraocular infections decreased significantly (p less than 0.0001) from 21 (0.69%) of 3059 during the first to 9 (0.11%) of 8179 intraocular operations during the second observation period . In the first period 11 vitrectomies and 2 enucleations due to bacterial endophthalmitis had to be performed . In the second period 2 vitrectomies and no enucleations were necessary (p less than 0.0001) . Our results indicate, that decontamination of the conjunctiva may be an import factor for the prevention of postoperative endophthalmitis following elective intraocular surgery. J Clin Microbiol, 1992 Mar, 30(3), 642 - 8 High levels of Gardnerella vaginalis detected with an oligonucleotide probe combined with elevated pH as a diagnostic indicator of bacterial vaginosis; Sheiness D et al.; We have demonstrated a new approach to diagnosing bacterial vaginosis (BV) that is based on measuring the concentration of Gardnerella vaginalis in vaginal fluid with DNA probes . G . vaginalis is virtually always present at high concentrations in women who have BV but is also detected frequently in normal women, usually at concentrations of less than 10(7) CFU/ml of vaginal fluid . Elevated vaginal pH is another sensitive indicator of BV, although it can occur in conjunction with other conditions . We have proposed that quantitative measurements of G . vaginalis using specific DNA probes can serve as a useful aid in diagnosing BV, provided the vaginal pH is above 4.5 . To test this hypothesis, a group of 113 women were first evaluated for BV by the standard set of clinical signs . Vaginal washes were collected, and aliquots were analyzed by quantitative culture for the concentration of G . vaginalis . Portions of these same samples were immobilized on nylon filters, along with standards for quantitation . The filters were incubated with a radiolabelled oligonucleotide specific for G . vaginalis 16S rRNA, and the subsequent autoradiographs were examined to determine levels of G . vaginalis in each sample . G . vaginalis at concentrations of greater than or equal to 2 x 10(7) CFU/ml and vaginal pH of greater than 4.5 were then analyzed for concurrence with the diagnoses based on clinical criteria . Results of this slot blot analysis gave a sensitivity of 95%, correctly categorizing 41 of 43 BV-positive specimens, and a specificity of 99%, correctly identifying 69 of 70 BV-negative specimens, compared with diagnosis based on clinical criteria. Mutat Res, 1992 Mar, 281(3), 157 - 61 Differential effect of the amino acid cystine in cultured mammalian and bacterial cells exposed to oxidative stress; Brandi G et al.; The effect of cystine in the cytotoxic response of cultured Chinese hamster ovary and Escherichia coli cells to challenge with hydrogen peroxide has been investigated . It was found that this amino acid could either protect or sensitize cells, depending on the cellular system . In fact, although a reduction in the growth-inhibitory effect of hydrogen peroxide was observed in mammalian cells, a marked increase in the susceptibility to oxidative stress was induced by cystine in bacteria . None of the amino acid precursors of glutathione, e.g., glutamate, glycine or cysteine, afforded protection in the mammalian cell system, whereas cysteine, but not glycine or glutamate, markedly sensitized bacteria to hydrogen peroxide-induced cell killing . In mammalian cells, methionine, an amino acid which is converted to cysteine, was also unable to modify the oxidative response . The results presented indicate that cystine displays differential effects in oxidatively injured mammalian or bacterial cells and suggest that the mechanism whereby the amino acid modulates the lethal action of hydrogen peroxide differs in the two cellular systems. Vet Immunol Immunopathol, 1992 Mar, 31(3-4), 241 - 53 Secretory activity of equine polymorphonuclear leukocytes: stimulus specificity and priming effects of bacterial lipopolysaccharide; Bochsler PN et al.; Neutrophil (PMN) contributions to the acute inflammatory process and host defense include generation of bioreactive oxygen metabolites and secretion of granule enzymes . We assessed equine PMN secretion using several PMN stimuli, singly and in combination with bacterial lipopolysaccharide (LPS) . LPS avidly associated with equine PMN, as shown by strong PMN labeling with FITC-conjugated LPS . LPS alone (1 or 10 micrograms ml-1) was a weak stimulus for PMN superoxide anion (O2-) generation, but preincubation with LPS followed by phorbol ester (PMA, 10 ng ml-1) significantly augmented (P less than 0.01) secretion of O2- (19.38 nmol O2- per 2 x 10(6) PMN per 5 min) over the amount generated by PMA stimulation alone (13.75 nmol O2-) . A qualitatively similar, but smaller O2(-)-generation response occurred when either opsonized zymosan or recombinant human C5a was used as the PMN stimulus . Arachidonic acid (ArA; 50-200 microM) was a potent stimulus, with secreted O2- levels similar to those from PMA-stimulated PMN . Preincubation of PMN with either the formyl peptide, fMLP, or platelet-activating factor before stimulation with ArA did not significantly increase O2- generation over levels obtained using ArA alone . Release of PMN granule enzymes was also quantitated . A small amount of lysozyme secretion resulted when PMN were exposed to LPS alone (8.20% of total cell content), and PMA stimulation caused marked release of PMN lysozyme (44.45%) . Non-specific proteolytic activity in PMN supernatants, assessed by cleavage of a collagen-rich substrate, was minimal with LPS as a sole stimulus (5.08%) . There was significant proteolytic activity (P less than 0.01) in supernatants from PMA-stimulated PMN (27.21%), and preincubation with LPS followed by PMA stimulation slightly enhanced (P less than 0.05) the release of PMN proteases (34.62%) . The activities of beta-glucuronidase, acid phosphatase, and alkaline phosphatase were minimal in PMN supernatants when using LPS and PMA as stimuli . The activity of PMN granule enzymes was found to be sensitive to the presence of normal equine serum, and proteolytic activity was markedly reduced (80.13% reduction) in the presence of 10% pooled serum. Arch Biochem Biophys, 1992 Mar, 293(2), 241 - 5 The bacterial hemoglobin from Vitreoscilla can support the aerobic growth of Escherichia coli lacking terminal oxidases; Dikshit RP et al.; Two Escherichia coli mutants that lack both cytochrome o and d terminal oxidases are able to grow with glucose as the carbon source but not with the aerobic substrates succinate or lactate . One of these, GV101, is a deletion mutant of cytochrome o and a point mutation of cytochrome d . The other, GK100, is a total deletion mutant of all the genes for both cytochromes . When these mutants were transformed with a plasmid containing the gene for the bacterial hemoglobin from Vitreoscilla, they were capable of growth in the presence of succinate or lactate and showed aerobic respiration in the presence of these substrates, unlike the parent strains . Cells transformed with a plasmid containing the gene for the hemoglobin but lacking the native promoter did not express the hemoglobin and did not respire . Membrane vesicles prepared from the cells consumed oxygen in the presence of succinate . This succinate-supported respiration decreased with successive washings of the vesicles but was restored by adding E . coli cytosol containing the hemoglobin or by adding the hemoglobin purified from Vitreoscilla . This respiration was inhibited by cyanide. Vet Rec, 1992 Feb 29, 130(9), 175 - 8 Effect of premilking teat disinfection on mastitis incidence, total bacterial count, cell count and milk yield in three dairy herds; Blowey RW et al.; An iodophor teat disinfectant was applied before milking by dip or spray to 50 cows and 50 cows were left untreated in each of three commercial herds . The mean incidence of clinical mastitis was reduced by 57 per cent, the total bacterial count by 70 per cent and the count of thermoduric organisms by 32 per cent . These results were not statistically significant, except that one herd showed a significant decrease in total bacterial count . There was no effect on somatic cell count, milk production or milk iodine residues . Atmospheric iodine concentrations increased in the two herds which applied the treatment as a spray, but the levels attained were not likely to be detrimental to human health. Eur J Biochem, 1992 Feb 15, 204(1), 93 - 8 The amino acid sequence of glutamate decarboxylase from Escherichia coli . Evolutionary relationship between mammalian and bacterial enzymes; Maras B et al.; The amino acid sequence of glutamate decarboxylase from Escherichia coli was solved by a combination of automated Edman degradation of peptide fragments derived by proteolytic and chemical cleavage and sequencing of DNA . Correct alignment of three peptides, for which no peptide overlaps were available, was achieved by sequencing a 1.1-kbp fragment of DNA produced by a polymerase-chain reaction using primers corresponding to sequences known to be in amino-terminal and carboxy-terminal regions of the protein . Sequence similarity (24% identity) with mammalian glutamate decarboxylase was found to be limited to a 55-residue sequence around the lysine residue that binds the coenzyme . Stronger similarity (38% identity), again confined to the same region, is seen with bacterial pyridoxal-phosphate-dependent histidine decarboxylase. Hybridoma, 1992 Feb, 11(1), 41 - 51 Enhanced transfection of a bacterial plasmid into hybridoma cells by electroporation: application for the selection of hybrid hybridoma (quadroma) cell lines; Bos R et al.; A procedure was investigated to transduce a bacterial plasmid containing a specific drug resistance marker (pSV2-neo), into a hybridoma cell line using electroporation . The effect of several buffers and the form of plasmid DNA (circular or linearized) on the stable transfection frequency were examined . When complete cell culture medium (DMEM) was used as electroporation buffer, we observed a two-fold increase in post-pulse viability and a ten- to thirty-fold increase in the transfection frequency of pSV2-neo, as compared with HEPES buffered 0.15 M sodium chloride . Supplementing DMEM with fetal bovine serum (DMEM + FBS) had some beneficial effect on post-pulse viability of the cells after electroporation, but did not markedly increase stable transfection frequency as compared with DMEM alone . Furthermore, with DMEM + FBS, the intact plasmid was transfected as effectively as linearized PSV2-neo . However, when using HEPES buffered saline, the transfection frequency of pSV2-neo increased two-fold after linearization as compared with intact plasmid . The drug resistance was used successfully as a marker for the selection of hybrid hybridoma (quadroma) cell lines after fusing two different hybridoma cell lines, producing anti-fibrin and anti-plasminogen activator antibodies respectively . The quadroma cells produced bispecific antibodies that are capable of accumulating plasminogen activator on a fibrin surface. Urol Clin North Am, 1992 Feb, 19(1), 25 - 34 Update on bacterial sexually transmitted disease; Zenilman JM; Sexually transmitted diseases (STDs) are the most frequently reported bacterial infections . In 1990, slightly more than 700,000 cases of gonococcal disease were reported to the Centers for Disease Control, and the estimated incidence of chlamydial infections is 3 million to 4 million annually . Bacterial genital ulcer diseases such as syphilis and chancroid are increasing in epidemic proportions . The author reviews bacterial STDs in two categories: exudative mucosal infections and genital ulcer diseases. Dig Dis Sci, 1992 Feb, 37(2), 248 - 56 Hepatobiliary excretion of bacterial formyl-methionyl peptides in rat . Structure activity studies; Anderson RP et al.; The bacterial chemotactic peptide formyl-met-leu-phe and its radioiodinated analog formyl-met-leu-{125I}tyr are rapidly excreted by the liver into bile following portal or systemic venous infusions in rats or after absorption from the gut lumen . To determine the molecular structural requirements for hepatobiliary excretion of formyl-methionyl peptides, structure-activity studies using portal venous infusions of 24 structural analogs of formyl-met-leu-tyr were performed in rats with biliary cannulae . Hepatic extraction of peptides was studied in vivo using external gamma counting after portal infusion . Efficient hepatobiliary excretion was not restricted to bioactive formyl peptides, but showed a broad specificity for different amino-acylated (formyl, acetyl, propionyl, carbobenzoxy) di- and tripeptides and no requirement for methionine in position one or for a free carboxy terminus . However, nonacylated peptides and an acyl-amino acid showed little excretion . Hepatic extraction of peptide was also related to N-acylation . Hepatic extraction and excretion of N-acyl peptides were also related to hydrophobicity . Thus, the presence of an N-acyl group is the key determinant of biliary excretion of inflammatory bacterial f-met peptides in the rat. Surg Gynecol Obstet, 1992 Feb, 174(2), 125 - 32 Effects of thromboxane synthetase inhibition on postburn mesenteric vascular resistance and the rate of bacterial translocation in a chronic porcine model; Tokyay R et al.; It is known that thromboxane (TX)B2, the metabolite of the potent vasoconstrictor TXA2, is elevated markedly in the serum of the patients immediately postburn . We had shown that extensive thermal injury causes a reduction in mesenteric blood flow that can lead to bacterial translocation from the intestine . In this study, we tested the hypothesis that the TX synthetase inhibitor, OKY-046, prevents increased mesenteric vascular resistance (MVR) and decreases the rate of translocation of bacteria seen after extensive thermal injury . Pigs in groups 1 (n = 6) and 2 (n = 6) had third degree burns of 40 per cent total body surface area under general anesthesia and were resuscitated according to the Parkland formula . Pigs in group 2 received 10 milligrams per kilogram of OKY-046 as a bolus just before the burn and 10 grams per kilogram per minute for 16 hours as a continuous infusion . Pigs in group 3 (control, n = 6) underwent general anesthesia only and received daily maintenance fluids of lactated Ringer's solution, 2 milliliters per kilogram per hour . OKY-046 prevented the significant increase in MVR seen during the first eight hours after burn . The total peripheral resistance (TPR) showed an early increase and a late decrease in the burn group, while the cardiac index (CI) and temperature (T) significantly increased after 24 hours . Administration of OKY-046 kept TPR, Cl, and T remarkably stable . OKY-046 reduced the rate of translocation of bacteria seen in the burn group from 67 to 17 per cent . Our results show that the blockade of thromboxane synthesis by OKY-046 prevented the early mesenteric vasoconstriction and the late hyperdynamic response seen after thermal injury and was useful in reducing the incidence of postburn translocation of bacteria. Int J Biochem, 1992 Feb, 24(2), 317 - 23 Bacterial lipopolysaccharide stimulates phospholipid synthesis and phosphatidylcholine breakdown in cultured human leukemia monocytic THP-1 cells; Chu AJ; 1 . De novo synthesis of phospholipid and its catabolism in human leukemia monocytic THP-1 cells were investigated . 2 . Radiolabelled precursors: {methyl-3H}chloride, {1,2-14C}ethanolamine and myo-{2-3H}inositol were readily incorporated into CHCl3-MEOH extractable lipid fraction as a function of time . 3 . The radiolabels derived from choline, ethanolamine and inositol were preferentially incorporated into PC, PE and PI fraction, respectively . The data indicate that de novo PL synthesis takes place, and the CDP-choline pathway is operative as a major pathway for PC synthesized in THP-1 cells . 4 . Bacterial endotoxin dose-dependently stimulated the incorporation of radiolabelled precursors . Approximately 50% stimulation in PC and PE synthesis was obtained in 20 hr, while the incorporation of {3H}inositol was rapidly stimulated by 170% within 4 hr, and the stimulation declined drastically thereafter . 5 . LPS did not alter the radiolabel distribution into PL in any of the three cases . 6 . In pulse-chase studies, the cells prelabelled with radioactive PL were exposed to LPS (1 micrograms/ml) . The breakdown of PC was enhanced about 30% within the first 2 hr followed by a stimulated PC synthesis observed in the next 4 hr . In contrast, LPS did not induce the hydrolysis of PE and PI . 7 . The data indicate that LPS produces a broad spectrum of stimulatory effects on PL synthesis and selectively stimulates the hydrolysis of PC via phospholipase C/D reaction in THP-1 cells. Infect Immun, 1992 Feb, 60(2), 596 - 601 In vitro inactivation of bacterial endotoxin by human lipoproteins and apolipoproteins; Emancipator K et al.; A chromogenic Limulus amebocyte lysate assay was used to measure the recovery of 1 endotoxin unit of endotoxin per ml . Purified human high-density lipoprotein, low-density lipoprotein, and apolipoprotein A1 (apo A1) at a maximum concentration of 1 g of protein per liter reduced the recovery to less than 40% of baseline in a both dose- and time-dependent manner and in the absence of other serum components . Furthermore, the lapine fever response to a dose of 1 ml of 5-ng/ml endotoxin per kg was reduced by greater than 0.5 degrees C (P less than 0.005) when the solution was preincubated in vitro with 0.5 g of apo A1 per liter . By the Limulus test, a maximum concentration of 0.01 g of apolipoprotein B (apo B) per liter (which contained deoxycholate, a known endotoxin-disaggregating agent) reduced recovery to 0% in a dose- but not time-dependent manner . In heat-inactivated (56 degrees C, 1 h) normal human serum, high-density lipoprotein cholesterol (P less than 0.005) and apo A1 (P less than 0.05) correlated inversely with endotoxin recovery, but, paradoxically, apo B correlated directly with endotoxin recovery (P less than 0.05), while low-density lipoprotein cholesterol showed no significant correlation . INTRALIPID alone had no effect on endotoxin recovery . Addition of a maximum of 10 g of INTRALIPID per liter to 0.0042 g of apo B per liter increased endotoxin recovery from approximately 30 to 80% (P less than 0.001), but addition of INTRALIPID to 0.25 g of apo A1 per liter decreased recovery from approximately 30 to 20% (P less than 0.001) . We conclude that (i) lipoproteins are endotoxin inactivators; (ii) this ability of lipoproteins may be modulated by their lipid component (lipid-endotoxin interaction); (iii) apo A1 is capable of directly inactivating endotoxin (protein-endotoxin interaction). Blood Coagul Fibrinolysis, 1992 Feb, 3(1), 19 - 23 Bacterial lipopolysaccharide induces phosphatidylcholine breakdown in human leukaemia monocytic U937 cells; Chu AJ et al.; We investigated the effect of bacterial lipopolysaccharide (LPS) on phospholipid (PL) turnover in human monocytic leukaemia U937 cells . Cells were pre-labelled with {3H}choline, {14C}ethanolamine and {3H}inositol for 24 h . By monitoring the radiolabel association with cellular PL, the data indicated that LPS (10 micrograms/ml) drastically altered the catabolism of choline-containing PL; it induced their breakdown by 50% within 20 min . The reutilization of choline or its phosphates for PL synthesis was also suggested as a result of regaining radiolabel in the next 40 min . Choline-containing PL then underwent a second degradation after 60 min; 50% decline in radiolabel was detected at 120 min . In contrast, LPS did not induce the breakdown of phosphatidylethanolamine and phosphatidylinositol through phospholipase C/phospholipase D (PLC/PLD) . No significant redistribution of the radiolabel in PL was detected in any cases during chasing . The data clearly indicate that LPS stimulates phosphatidylcholine breakdown, implying that the liberation of phosphatidic acid or diacylglycerol via PLC/PLD reaction may be relevant to the initiation of LPS-induced monocytic activation. Jpn J Antibiot, 1992 Feb, 45(2), 123 - 35 {Therapeutic evaluation of imipenem/cilastatin sodium for bacterial infections in patients with hematological diseases}; Sawae Y et al.; 1 . To evaluate the efficacy and tolerance of imipenem/cilastatin sodium (IPM/CS) in severe infections associated with hematopoietic disorders, IPM/CS was administered to a total of 105 patients . 2 . Out of 96 patients evaluable for efficacy, clinical responses were excellent in 23 patients, good in 30, fair in 15, poor in 19 and unknown in 9, and the overall response rate was 60.9% . 3 . The most common underlying hematopoietic disease was acute non-lymphocytic leukemia and the most common infections were sepsis and suspected sepsis . 4 . Daily dose, severity of infection and neutrophil count had effects on the clinical response . 5 . The overall eradication rate of bacteria was 83.7% . 6 . Side effects were observed in 10 patients (9.5%) and abnormal laboratory test results in 12 (11.4%) . From the above findings, we have concluded that IPM/CS is very useful for the treatment of severe infections in compromised patients with hematopoietic diseases. Appl Environ Microbiol, 1992 Feb, 58(2), 754 - 7 Detection of low numbers of bacterial cells in soils and sediments by polymerase chain reaction; Tsai YL et al.; Polymerase chain reaction was used to amplify the low copy number of two 16S ribosomal gene fragments from soil and sediment extracts . Total DNA for polymerase chain reaction was extracted from 1 g of seeded or unseeded samples by a rapid freeze-and-thaw method . Amplified DNA fragments can be detected in DNA fractions isolated from seeded soil containing less than 3 Escherichia coli cells and from seeded sediments containing less than 10 cells . This research demonstrated that coupling polymerase chain reaction to direct DNA extraction improves sensitivity by 1 and 2 orders of magnitude for sediments and soils, respectively . This technique could become a powerful tool for genetic ecology studies. Nippon Jinzo Gakkai Shi, 1992 Feb, 34(2), 181 - 9 {Survival of bacteria, and release of the endotoxin from the bacterial cells in the dialysates}; Koga N et al.; Survival of bacteria and release of the endotoxin from the bacteria with and without ultraviolet irradiation in three kinds of dialysate were investigated . The results obtained are as follows: (1) No growth of S . aureus, E . coli, P . aeruginosa, Aspergillus and C . albicans in the saturated dialysates tested were observed . (2) All of the bacterial cells tested here is gradually, and naturally spontaneously inactivated in all the dialysates . (3) Among the dialysates tested, the saturated dialysate, AF-2, is the most effective for inactivating P . aeruginosa ATCC, but the effect depends upon the isolates of P . aeruginosa . (4) The inactivating effect was somewhat decreased when the saturated AF-2 solution was diluted, but the killing effect was still maintained . (5) The bacterial cells are constantly and significantly inactivated by UV irradiation, especially by the direct irradiation . The indirect irradiation, i.e., through glass, has remarkably less effective than the direct one . However, a tendency of the decrease of bacterial cells by the indirect irradiation is maintained with the killing effect of the dialysated, especially in the case of AF-2 solution . (6) No significant increase of endotoxin was observed, even when the bacterial cells were killed by UV irradiation . Therefore, it is recommended to use UV irradiation for inactivating the bacteria . From the results obtained here, it is indicated that there is no possibility of the growth of naturally contaminated bacteria in the dialysates, and is an effectiveness of the use of UV irradiation for inactivating the bacteria cells, in terms of release of the endotoxin from the dead cells. Acta Neurol (Napoli), 1992 Feb, 14(1), 6 - 9 Cerebrospinal fluid interleukin-6 and IgE in bacterial and viral meningitis; Perrella O et al.; The aim of the present study was to assess the significance of IgE and interleukin-6 (IL-6) in paired CSF and serum of patients with viral and bacterial infections of the central nervous system . The results suggest that the detection of IL-6 and IgE in CSF is an useful marker for monitoring course and prognosis of these patients. Int J Exp Pathol, 1992 Feb, 73(1), 95 - 8 The HID50 (hypothermia-inducing dose 50): an alternative to the LD50 for measurement of bacterial virulence; Soothill JS et al.; A group of 40 mice involved in bacterial LD50 estimations was monitored over a period of 4 days by two teams of observers . One team measured the colonic temperature of the mice, the other assessed them clinically and killed those they judged terminally ill . The temperatures of all mice considered terminally ill dropped before this judgement was made . Measurement of such infection-induced hypothermia provides a more objective, reproducible, and earlier endpoint than that used in conventional LD50 estimation. J Otolaryngol, 1992 Feb, 21(1), 48 - 53 Hospitalized croup (bacterial and viral): the role of rigid endoscopy; Tan AK et al.; This is a retrospective study of 500 cases of hospitalized patients with the diagnosis of croup (laryngotrachitis), admitted between January 1986 and August 1988, at the Montreal Children's Hospital . The patient's age, sex, clinical history, physical examination, number of admissions, season of admission, method of diagnosis, treatment and management were reviewed . Two per cent of these patients were diagnosed as bacterial croup . All of them required intubation and endoscopic evaluation in their management . Of all the viral croup patients, less than one-third were severe enough to require an intensive care setting for their management . In these patients, 6% required endotracheal intubation, and on endoscopy, a significant number of these patients had an endoscopic airway abnormality in addition to croup (subglottic edema) . According to our findings, we suggest diagnostic micro-laryngoscopy and bronchoscopy be performed on certain groups of croup patients because of their higher yield of airway abnormality on endoscopy. Mol Microbiol, 1992 Feb, 6(4), 443 - 9 Evolutionary origins of bacterial bioluminescence; O'Kane DJ et al.; In bacteria, most genes required for the bioluminescence phenotype are contained in lux operons . Sequence alignments of several lux gene products show the existence of at least two groups of paralogous products . The alpha- and beta-subunits of bacterial luciferase and the non-fluorescent flavoprotein are paralogous, and two antennae proteins (lumazine protein and yellow fluorescence protein) are paralogous with riboflavin synthetase . Models describing the evolution of these paralogous proteins are suggested, as well as a postulate for the identity of the gene encoding a protobioluminescent luciferase. J Appl Bacteriol, 1992 Feb, 72(2), 139 - 45 Comparison of definitions of the lag phase and the exponential phase in bacterial growth; Zwietering MH et al.; Different definitions for the lag time and of the duration of the exponential phase can be used to calculate these quantities from growth models . The conventional definitions were compared with newly proposed definitions . It appeared to be possible to derive values for the lag time and the duration of the exponential phase from the growth models and differences between the various definitions could be quantified . All the different values can be calculated from the growth parameters microm, lambda and alpha . Therefore, it appeared to be unnecessary to use complicated mathematical equations; simple equations were adequate . For the Gompertz model the conventional definition of the lag time did not differ appreciably from the newly proposed definition . The end-point of the exponential phase and thus the duration of the exponential phase differed considerably for the two definitions . For the logistic model the two definitions lead to considerable differences for all quantities . It is recommended that the conventional definition is used for calculating the lag time . For the duration of the exponential phase it is recommended that the new definition is used . The value can be calculated, however, directly from the conventional growth parameters. Am J Physiol, 1992 Feb, 262(2 Pt 1), G312 - 8 {15N}- and {14C}glutamine fluxes across rabbit ileum in experimental bacterial diarrhea; Nath SK et al.; L-Glutamine (Gln) fluxes and the effects of Gln on Na and Cl transport were studied across the ileum of healthy and rabbit diarrheagenic Escherichia coli (RDEC-1)-infected weanling rabbits . Stable ({alpha-15N}Gln) and radioisotopic ({U-14C}Gln) tracers provided identical estimates of Gln transport both in healthy (H) and infected (I) rabbits . RDEC-1 infection, however, decreased net Gln flux {Jnet{14C}Gln = 682 +/- 147 (H) vs . 278 +/- 63 (I); Jnet{15N}Gln = 739 +/- 160 vs . 225 +/- 110 nmol.h-1.cm-2} due to a reduction in mucosal-to-serosal flux . After addition of Gln, increases in net Na absorption {delta Jnet{15N}Gln = 1.87 +/- 0.45 (H) vs . 0.70 +/- 0.27 (I) microeq.h-1.cm-2} and short-circuit current (delta Isc) {1.80 +/- 0.40 (H) vs . 0.74 +/- 0.14 (I) microeq.h-1.cm-2} were also reduced in infected rabbits . Addition of glucose after Gln, however, stimulated Na absorption further . These results indicate that 1) Gln is actively absorbed as intact Gln molecule across rabbit ileum; 2) Gln stimulates an electrogenic Na absorption in a 1:2 ratio that may be further stimulated by glucose; and 3) in RDEC-1 infection electroneutral NaCl absorption, intact Gln absorption, and electrogenic stimulation of Na absorption by glutamine are reduced. Yakugaku Zasshi, 1992 Feb, 112(2), 129 - 34 {Antipyretic effects of traditional Chinese medicines in bacterial endotoxin-induced febrile rabbits}; Itami T et al.; The antipyretic effects of oral administration of eight traditional Chinese medicines (dried extracts) were tested in febrile rabbits injected with bacterial pyrogen, lipopolysaccharide (LPS) 0.05 micrograms/kg (i.v.) . The traditional Chinese medicines were given 0.6-2 g/10 ml/kg (p.o.) simultaneously with LPS . The most potent antipyretic effect was observed in Dai-jyoki-to (Ta-chen-chi-tang) . The moderate antipyretic effects were observed in Toki-syakuyaku-san (Tang-kuei-shao-yao-san) and Syo-saiko-to (Hssiao-chai-hu-tang) . Koso-san (Hsiang-su-san), Oren-gedoku-to (Huang-lien-chieh-tu-tan), Gorei-san (Wu-ling-san), Kakkon-to (Ko-ken-tang) and Byakkoka-ninjin-to (Pai-hu-chia-jen-sheng-tang) showed no effects. Lik Sprava, 1992 Feb, (2), 53 - 7 {A structural analysis of hemato-encephalic barrier function in bacterial meningoencephalitis}; Iarosh OA; A correlation analysis of the content of metals in the cerebrospinal fluid in 70 patients with bacterial meningoencephalitis showed that an increase of permeability of microvessel walls for macro- and microelements is accompanied by a reduction of the regulating activity of barrier structures . Pathomorphological examination of the brain by the method of scanning electron microscopy revealed structural changes of the microvessel walls as the basis of disorders of the barrier-transport function. Protein Expr Purif, 1992 Feb, 3(1), 41 - 9 Rice cystatin: bacterial expression, purification, cysteine proteinase inhibitory activity, and insect growth suppressing activity of a truncated form of the protein; Chen MS et al.; A cDNA clone that encodes oryzacystatin, a cysteine protease inhibitor from rice, was isolated and expressed in Escherichia coli BL-21 (DE3) using an expression plasmid under the control of a T7 RNA polymerase promoter . The construct pT7OC 9b encoded a fusion protein containing 11 amino acid residues of the NH2 terminus of the bacterial protein phi 10 and 79 residues of oryzacystatin lacking 23 NH2-terminal residues of the wild-type protein . Recombinant oryzacystatin (ROC) constituted approximately 10% of the total bacterial protein mass and was purified in a single step by anion-exchange chromatography . The inhibitory activity of ROC toward papain (Ki = 3 x 10(-8) M) was comparable with that of the naturally occurring protein isolated from rice . Caseinolytic activity in midgut homogenates from seven species of stored product insects was inhibited from 18 to 85% by ROC, whereas the same activity was inhibited from 14 to 69% by the serine proteinase inhibitor phenylmethylsulfonyl fluoride . Midguts of stored product insects apparently contain both cysteine proteinases and serine proteinases, but the relative amounts vary with the species . When fed to the red flour beetle, Tribolium castaneum, 10 wt% ROC in the diet suppressed growth approximately 35% relative to that of the control group of insects. Res Microbiol, 1992 Feb, 143(2), 199 - 209 Heterogeneity in restriction patterns of Gardnerella vaginalis isolates from individuals with bacterial vaginosis; Nath K et al.; This study was undertaken to resolve the genetic make up of Gardnerella vaginalis present in bacterial vaginosis (BV) . DNA from several G . vaginalis isolates from within and between individual BV patients were compared by BamHI, ClaI and EcoRI restriction endonuclease analysis (REA) followed by a restriction fragment length polymorphism (RFLP) study, utilizing a 5.7-kb BamHI G . vaginalis ATCC14018 DNA probe . Four G . vaginalis isolates from one patient (GVP-062) were composed of 3 different biotypes (biotypes 3, 5 and 8), and while the REA mirrored the biotype, in RFLP studies at least 3 isolates had DNA fragments in common . All of the isolates from 2 other patients (GVP-063 and GVP-072) represented a single biotype (biotype 2), but under REA and in RFLP studies, the isolates GVP-063 differed from GVP-072 . An opposite case existed with the isolates GVP-072 (biotype 2) and GVP-065 (biotype 5), which appeared similar under REA and in RFLP studies . Finally, reisolates after 8 weeks (GVP-080) from a BV patient (isolates GVP-065) representing the same biotype (biotype 5) differed under REA and in RFLP studies . Thus, lacking any unique DNA fingerprint, G . vaginalis occurring in BV represents a (genetically) mixed population. Mol Microbiol, 1992 Feb, 6(4), 435 - 42 Porins and specific channels of bacterial outer membranes; Nikaido H; Porins and specific channels both produce water-filled pores that allow the transmembrane diffusion of small solutes, but the latter contain specific ligand-binding sites within the channels . Recent structural studies show that many or most of these proteins exist as beta-barrels with the beta-strands traversing the thickness of the outer membrane . The channels often have diameters in the range of 1 nm, and thus the penetration rates of solutes through porin channels are likely to be affected strongly by what appear to be minor differences in the size, shape, hydrophobicity or charge of the solute molecule . With the specific channels, the presence of binding sites can accelerate very significantly the diffusion of some ligands when they are present at low concentrations . Thus these simple channels can sometimes achieve a surprising degree of real or apparent specificity . Recent data tend to favour the idea that these proteins are first exported into the periplasm, and then inserted into the outer membrane . Although lipopolysaccharides seem to play a significant role in the final assembly of the trimeric porins, the details of the targeting process still remain to be elucidated. Mol Microbiol, 1992 Feb, 6(4), 425 - 33 Control of bacterial DNA supercoiling; Drlica K; Two DNA topoisomerases control the level of negative supercoiling in bacterial cells . DNA gyrase introduces supercoils, and DNA topoisomerase I prevents supercoiling from reaching unacceptably high levels . Perturbations of supercoiling are corrected by the substrate preferences of these topoisomerases with respect to DNA topology and by changes in expression of the genes encoding the enzymes . However, supercoiling changes when the growth environment is altered in ways that also affect cellular energetics . The ratio of {ATP} to {ADP}, to which gyrase is sensitive, may be involved in the response of supercoiling to growth conditions . Inside cells, supercoiling is partitioned into two components, superhelical tension and restrained supercoils . Shifts in superhelical tension elicited by nicking or by salt shock do not rapidly change the level of restrained supercoiling . However, a steady-state change in supercoiling caused by mutation of topA does alter both tension and restrained supercoils . This communication between the two compartments may play a role in the control of supercoiling. Prog Urol, 1992 Feb, 2(1), 93 - 8 {Non-specific bacterial ureteritis: a rare entity}; Jichlinski P et al.; Based on a recent case of non-specific bacterial ureteritis, the authors review the clinical features of what has now become a rare disease . They also evaluate the role of computed tomography in the differential diagnosis of this disease. Nutr Clin Pract, 1992 Feb, 7(1), 31 - 6 Limiting bacterial contamination of enteral nutrient solutions: 6-year history with reduction of contamination at two institutions; Fagerman KE; Running enteral nutrient solution (ENS) bacterial contamination logs that were collected over a 6-year time frame are presented . At hospital A, reconstituted ENS were prepared by the pharmacy department in bulk and were frozen in the final container . These solutions were cultured and counted after 12 hours of storage at room temperature to stimulate this institution's hang time . Major reductions in ENS bacterial counts occurred after improvements in sanitation, a reduction in solution hang time, the conversion to the use of sterile water for dilution and reconstitution of ENS, and most dramatically, after the incorporation of a preservative (potassium sorbate) to reconstituted ENS . At hospital B, ENS usage consisted of canned feedings that were prepared by nursing personnel and were transferred to the feeding container at the bedside . Reductions in final counts of contamination of ENS occurred after procedural changes, which included container changes every 24 hours, use of sterile water for dilution, cleansing of can lids with alcohol swabs before use, rinsing and air drying of intermittent feeding containers between feedings, and limiting feeding container fills to 4-hour hang time quantities . At both institutions, the value of an ongoing three-class enteral quality control program with a defined acceptance/rejection criteria was demonstrated in that ENS contamination was reduced to acceptable levels comparable with federal standards for milk and dairy products. MMWR Morb Mortal Wkly Rep, 1992 Jan 24, 41(3), 36 - 7 Bacterial contamination of platelet pools--Ohio, 1991; Phosphorylation of bacterial response regulator proteins by low molecular weight phospho-donors; Department of Molecular Biology, Princeton University, NJ 08544Bacterial motility and gene expression are controlled by a family of phosphorylated response regulators whose activities are modulated by an associated family of protein-histidine kinases . In chemotaxis there are two response regulators, CheY and CheB, that receive phosphoryl groups from the histidine kinase, CheA . Here we show that the response regulators catalyze their own phosphorylation in that both CheY and CheB can be phosphorylated in the complete absence of any auxiliary protein . Both CheY and CheB use the N-phosphoryl group in phosphoramidate (NH2PO3(2-)) as a phospho-donor . This enzymatic activity probably reflects the general ability of response regulators to accept phosphoryl groups from phosphohistidines in their associated kinases . It provides a general method for the study of activated response regulators in the absence of kinase proteins . CheY can also use intermediary metabolites such as acetyl phosphate and carbamoyl phosphate as phospho-donors . These reactions may provide a mechanism to modulate cell behavior in response to altered metabolic states. Proc Natl Acad Sci U S A, 1992 Jan 15, 89(2), 613 - 7 Femtosecond spectral evolution of the excited state of bacterial reaction centers at 10 K; Vos MH et al.; The femtosecond spectral evolution of reaction centers of Rhodobacter sphaeroides R-26 was studied at 10 K . Transient spectra in the near infrared region, obtained with 45-fs pulses (pump pulses centered at 870 nm and continuum probe pulses), were analyzed with associated kinetics at specific wavelengths . The t = 0-fs transient spectrum is very rich in structure; it contains separate induced bands at 807 and 796 nm and a bleaching near 760 nm, reflecting strong changes in interaction between all pigments upon formation of the excited state . A complex spectral evolution in the 800-nm region, most notably the bleaching of the 796-nm band, takes place within a few hundred femtosecond--i.e., on a time scale much faster than electron transfer from the primary donor P to the bacteriopheophytin acceptor HL . The remarkable initial spectral features and their evolution are presumably related to the presence of HL, as they were not observed in the DLL mutant of Rhodobacter capsulatus, which lacks this pigment . A simple linear reaction scheme with an intermediate state cannot account for our data; the initial spectral evolution must reflect relaxation processes within the excited state . The importance for primary photochemistry of long distance interactions in the reaction center is discussed. Commun Dis Rep CDR Rev, 1992 Jan 3, 2(1), R7 - 9 Bacterial infection and human cancer: association or causation? Caygill CP. Environmental factors such as diet, industrial pollution, smoking and viruses account for some 80% of cancers and more than one-quarter of all non-accidental deaths . Little attention has been paid to the relationship between bacterial infection and cancer but there is growing evidence to support this link . For some sites, such as stomach and colorectum, the evidence is relatively strong, whereas for others, such as biliary tract and bladder, it is more circumstantial. Am J Obstet Gynecol, 1992 Jan, 166(1 Pt 1), 100 - 3 Incidence of pelvic inflammatory disease after first-trimester legal abortion in women with bacterial vaginosis after treatment with metronidazole: a double-blind, randomized study; Larsson PG et al.; OBJECTIVE: The purpose of this study was to evaluate the effect of metronidazole treatment on the incidence of postoperative pelvic inflammatory disease after first-trimester abortion in women with bacterial vaginosis . STUDY DESIGN: A double-blind, randomized, multicenter study was conducted on 231 women undergoing first-trimester legal abortion and fulfilling the criteria for bacterial vaginosis . The women were randomized to either metronidazole 500 mg three times daily for 10 days or placebo . Treatment was started at the outpatient visit the week before the operation . RESULTS: Among the 174 women who could be evaluated, pelvic inflammatory disease developed in 14 after the abortion . In the treatment group there were three infections (3.8%) compared with 11 (12.2%) in the placebo group (p less than 0.05) . CONCLUSION: These data suggest that patients with bacterial vaginosis should be treated in conjunction with first-trimester abortion because treatment with metronidazole reduces the postoperative infection rate more than three times. Plant Mol Biol, 1992 Jan, 18(1), 47 - 53 A plant signal sequence enhances the secretion of bacterial ChiA in transgenic tobacco; Lund P et al.; When the secreted bacterial protein ChiA is expressed in transgenic tobacco, a fraction of the protein is glycosylated and secreted from the plant cells; however most of the protein remains inside the cells . We tested whether the efficiency of secretion could be improved by replacing the bacterial signal sequence with a plant signal sequence . We found the signal sequence and the first two amino acids of the PR1b protein attached to the ChiA mature protein directs complete glycosylation and secretion of the ChiA from plant cells . Glycosylation of this protein is not required for its efficient secretion from plant cells. J Cell Physiol, 1992 Jan, 150(1), 204 - 13 Regulation of NF-kappa B activity in murine macrophages: effect of bacterial lipopolysaccharide and phorbol ester; Vincenti MP et al.; Nuclear factor kappa-B (NF-kappa B) has been shown to play an important role in LPS-mediated induction of several genes in macrophages . Several studies have implicated protein kinase C (PKC) or cAMP-dependent protein kinase in the regulation of NF-kappa B activity . In this study we have investigated the mechanism of NF-kappa B induction in murine macrophages . A chloramphenicol acetyl transferase (CAT) expression vector containing multiple copies of the TNF-alpha NF-kappa B element was transfected into the RAW264 macrophage-like cell line and assessed for inducible CAT activity . LPS treatment of the transfected cells resulted in a significant induction of CAT activity . CAT activity was not induced by treatment with phorbol myristate acetate (PMA) or the cAMP analogue 8-bromo cAMP . To further study NF-kappa B induction, nuclear extracts were prepared from RAW264 cells . Extracts from RAW264 cells that were treated from 30 min to 2 hr with LPS had a significant increase in NF-kappa B binding activity as determined by the electrophoresis mobility shift assay (EMSA) . Treatment of these cells from 30 min to 2 hr with PMA did not result in such binding activity . U.V . crosslinking analysis of the DNA-binding activity confirmed these results and indicated that LPS induced a 55 KD DNA-binding protein . Induction of this NF-kappa B binding activity was not inhibited by pretreatment with the PKC inhibitor H-7 . H-7 did inhibit induction of TPA responsive element binding by either LPS or PMA . Prolonged exposure to phorbol ester, a treatment which down-regulates PKC, had no effect on LPS induction of NF-kappa B activity in these cells . These results suggest that the induction of NF-kappa B in macrophages by LPS is independent of PKC. Proc Natl Acad Sci U S A, 1992 Jan 1, 89(1), 33 - 7 Transfer of the bacterial gene for cytosine deaminase to mammalian cells confers lethal sensitivity to 5-fluorocytosine: a negative selection system; Mullen CA et al.; Expression of the bacterial gene for cytosine deaminase (CD; EC 3.5.4.1) in mammalian cells was evaluated as a negative selection system or suicide vector for potential use in gene transfer studies and therapies . Mammalian cells, unlike certain bacteria and fungi, do not contain the enzyme CD and do not ordinarily metabolize cytosine to uracil . Nor do they metabolize the innocuous compound 5-fluorocytosine to the highly toxic compound 5-fluorouracil . The Escherichia coli CD gene underwent PCR oligonucleotide-directed mutagenesis to enhance its expression in a eukaryotic system and it was then cloned into an expression vector, pLXSN, that also contains a neomycin-resistance gene . Murine fibroblast lines were transfected with the plasmid and subjected to brief selection in the neomycin analogue G418 . Lysates from these cell populations exhibited significant CD activity detected by conversion of radiolabeled cytosine to uracil . In clonogenic assays transfected cells expressing CD were selectively killed by incubation in 5-fluorocytosine, whereas control cell lines were not . Dose-response studies evaluating {3H}thymidine incorporation or cloning efficiency demonstrated profound inhibition at and above 65 micrograms of 5-fluorocytosine per ml . Mixed cellular assays showed that CD-positive cells could be eliminated without bystander killing of other cells . Retrovirus-mediated CD gene transfer into various tissues was also demonstrated . Thus CD, with its ability to produce the toxic antimetabolite 5-fluorouracil from 5-fluorocytosine, may be useful as a negative selection system for studies and treatments employing gene transfer techniques. Arch Intern Med, 1992 Jan, 152(1), 99 - 104 IgG subclass deficiency and sinopulmonary bacterial infections in patients with alcoholic liver disease; Spinozzi F et al.; Abnormalities in IgG subclass distribution were sought in serum samples and bronchoalveolar lavage fluid from 15 patients with alcoholic liver disease to explain their increased susceptibility to bacterial respiratory infections . Serum IgG4 deficiency alone or in association with low IgG2 levels was revealed in approximately 30% of patients with alcoholic liver disease . This fact prompted us to further investigate the immunoglobulin concentrations in broncho-alveolar lavage fluid, paying special attention to the distribution of IgA and IgG subclasses . IgA levels were found to be normal or slightly elevated . However, there were substantial defects in total IgG and IgG1 concentrations, often associated with reduced IgG2 and IgG4 levels, in approximately 70% of patients with alcoholic liver disease, which proved to be closely correlated with the number and type (pneumonia) of bacterial respiratory infections . A prospective study of intravenous immunoglobulin substitutive therapy involving two patients with recurrent pneumonia and very low serum IgG2 values demonstrated a reduction in the number of respiratory infectious episodes as well as an increase in both serum and, to a lesser extent, bronchoalveolar lavage fluid IgG1 and IgG2 levels . We identified immune defects that may represent an important pathogenetic mechanism that, when considered together with the alcohol-related suppression of alveolar macrophage and ciliary functions and the inhibition of leukocyte migration into the lungs, should help clarify the complex relationships between alcohol and immune defense. J Thorac Cardiovasc Surg, 1992 Jan, 103(1), 124 - 8; discussion 128-9 Mitral valve repair for bacterial endocarditis; Hendren WG et al.; Twenty-two patients with mitral insufficiency resulting from native valve endocarditis underwent mitral valve repair . Six patients had acute endocarditis with positive blood cultures and active valve infection . Sixteen patients were cured of active infection, but mitral insufficiency developed as a result of prior infection . Mean age was 48.5 +/- 21.7 years; 13 (59%) were male . Mean New York Heart Association functional class was 2.6 +/- 1.2 . Multiple valve lesions were present in 11 (50%) patients . Valve abnormalities included leaflet perforation in 13 patients, chordal rupture or elongation in 14, vegetations in 5; and annular abscess in 1 . In patients with acute endocarditis all macroscopically infected tissue was excised . Multiple techniques were required to achieve valve competence . Suture or patch closure of perforation was done in 14 patients, chordal shortening or transfer in 9, leaflet resection and closure in 4, leaflet resection with pericardial patching in 5, and annuloplasty in 15 . Mitral valvuloplasty was combined with other procedures in 11 (50%) patients . There were two (9%) hospital deaths, both occurring in patients with healed endocarditis . There was one (9%) death in a patient undergoing an isolated procedure and one (9%) in a patient undergoing a combined procedure . Mean follow-up was 24 +/- 16.8 months and was complete . Seventeen (85%) were in New York Heart Association functional class I, and three (15%) were in class II . There were no late deaths, reoperations, recurrent endocarditis, thromboembolic events, or other valve-related morbidity . We conclude that mitral valve repair for insufficiency resulting from bacterial endocarditis (1) is possible in acute and healed disease, (2) has a low operative mortality, and (3) has resulted in patients free of recurrent infection and valve-related morbidity and mortality . Mitral valve repair is an attractive alternate to valve replacement in bacterial endocarditis. Am J Gastroenterol, 1992 Jan, 87(1), 28 - 30 H . pylori, the most common bacterial infection in Africa: a random serological study; Holcombe C et al.; This study was carried out to determine the prevalence of antibodies to Helicobacter pylori in northern Nigeria, a region with a low incidence of peptic ulceration . In a random, serological survey of 268 subjects, 228 (85%) of the population studied had IgG antibodies to H . pylori . Fifty-eight of these subjects had experienced dyspepsia in the preceding 6 months . The majority of the population (82%) is infected between the ages of 5 and 10 . Despite the high prevalence of antibodies to H . pylori, peptic ulcer is uncommon, suggesting that H . pylori is not important in the etiology of peptic ulcer in this population . Indeed, most patients infected by H . pylori are asymptomatic . The possible reasons for this are discussed. Arch Biochem Biophys, 1992 Jan, 292(1), 266 - 72 Kinetics of inhibition of peptide bond formation on bacterial ribosomes; Theocharis DA et al.; A cell-free system derived from Escherichia coli has been used in order to study the kinetics of inhibition of peptide bond formation with the aid of the puromycin reaction in solution . A similar study has been carried out earlier on a solid support matrix with the same inhibitors . We find that the overall pattern of the kinetics of inhibition is the same in the two systems . At low concentrations of inhibitor there is a competitive phase of inhibition, whereas at higher concentrations of inhibitor the type of inhibition becomes mixed noncompetitive . The values of Ki of the competitive phase in the system in solution are: 5.8 microM (amicetin), 0.2 microM (blasticidin S), 0.5 microM (chloramphenicol), and 0.5 microM (tevenel) . The inhibitors amicetin, blasticidin S, and tevenel interact with the ribosome in a reaction which is slower than that of the substrate puromycin, showing clear-cut characteristics of slow-onset inhibition in both systems . Chloramphenicol, on the other hand does not easily show such a delay in solution . It interacts with the ribosome relatively faster than the other three antibiotics . Despite this, chloramphenicol too shows characteristics of time-dependent inhibition. Med Clin North Am, 1992 Jan, 76(1), 173 - 203 Medical management of AIDS patients . Bacterial and fungal infections; Daar ES et al.; The rapid and thus far generally inexorable rise in HIV infections has led to a series of opportunistic infection that includes those caused by bacteria, yeasts, and members of the Eumycetes . The infections range in prevalence from occasional to highly prevalent, in severity from trivial to fatal, and in anatomic areas involved from local to disseminated . They occur as isolated, concurrent, or sequential infections with regard to other opportunistic diseases . Some vary in their geographic distribution . They may be newly acquired or reactivated and occur early or late in the course of HIV infection . Bacterial infections are usually easily treated, although they frequently disseminate and often recur after seemingly appropriate treatment . In contrast, all but the mildest fungal infections are difficult to treat and even more difficult or impossible to eradicate . The diagnosis of bacterial and fungal infections begins with clinical suspicion and involves relatively standard methodology . Treatment of the systemic mycoses and some bacterial infections in HIV infected patients is punctuated by exaggerated side effects of therapy, frequent relapses, and the need for maintenance suppressive therapy. Scand J Infect Dis, 1992, 24(2), 189 - 95 Loss of erythrocyte complement receptors (CR1; CD35) in patients with acute episodes of septicaemia or bacterial meningitis; Olesen B et al.; 14 patients with septicaemia or bacterial meningitis were examined for serum levels of erythrocyte complement receptors CR1 (C3b/C4b; CD35) by an enzyme-linked immunosorbent assay and levels of circulating immune complexes (IC) by a polyethylene glycol precipitation complement consumption method, and concentrations of complement C3d split products in plasma by intermediate gel rocket immunoelectrophoresis . The CR1 receptor levels were significantly lower on day 7-8 after admission than on day 1-2 (p = 0.01), and than the levels 3-4 months later (p = 0.002) . Both the levels of IC in serum and the C3d concentrations were significantly higher on day 7-8 than 3-4 months later (p = 0.02 and p = 0.004) . The mechanism behind a temporary decreased expression of CR1 in patients with acute episodes of septicaemia or purulent meningitis is not fully known, and further investigations are needed to clarify whether this reduction in erythrocyte CR1 causes an impaired disposal of IC in patients with infections. Biotech Histochem, 1992 Jan, 67(1), 27 - 9 Rapid identification of viable bacterial spores using a fluorescence method; Sharma DK et al.; A method has been devised to differentiate viable and nonviable bacterial spores . "Germination-like" changes are initiated in spores with performic acid and lysozyme . The germinated spores are stained with aqueous acridine orange, a fluorescent dye . Nonviable spores fluoresce lemon-green and viable spores orange-red . It is proposed that with the use of a membrane filter resistant to performic acid and lysozyme, the method may be used for spore enumeration in foods in about 4 hr compared to conventional plating methods, which usually require up to 72 hr. Int Arch Allergy Immunol, 1992, 97(1), 78 - 82 Effect of cow's milk protein absorption on the anaphylactic and systemic immune responses of young rabbits during bacterial diarrhoea; Gotteland M et al.; In vivo and in vitro studies recently showed that intestinal permeability to cow's milk proteins such as beta-lactoglobulin increased transiently in infant rabbits infected by the enteroadherent Escherichia coli strain RDEC-1 at weaning . The consequences of this enhanced permeability for local anaphylactic and/or systemic immune responses were studied in infected and age-matched control rabbits at weaning, given either water or cow's milk diluted in water in addition to their solid diet . The systemic immune response was determined by measuring IgG, IgA, IgM, and IgE antibodies to cow's milk proteins . A rise in IgG and IgM, but not IgA or IgE, antibody titres to milk protein was observed . Further, infected milk-drinking rabbits had significantly higher beta-lactoglobulin and casein IgG titres than milk-drinking controls . The anaphylactic response was tested by mounting ileal segments in Ussing chambers, adding the milk proteins to the serosal side of the tissue and recording the variation in short-circuit current in order to detect any electrogenic chloride secretion . No change in short-circuit current was observed in the presence of milk proteins in infected or age-matched control milk-drinking rabbits, indicating the absence of any immediate hypersensitivity reaction . These results indicate that at weaning, the rabbit is not prone to developing anaphylactic responses to milk proteins . The rise in milk protein IgG antibodies that followed infection with RDEC-1 might favour the elimination of milk protein-IgG immune complexes from the systemic circulation . The present results emphasize the evidence that genetic background and/or animal species greatly influence the immune response to an antigenic load. Indian J Lepr, 1992 Jan-Mar, 64(1), 14 - 27 Reaction of peripheral nerves to vascular and bacterial injuries; Rasouli I et al.; Mouse sciatic nerves were subjected to devascularization, M . leprae inoculation, and combined insult of devascularization + footpad inoculation (FPI) . Changes were seen in FPI nerves only after eight months, but in cases of combined insult, changes were evident in hours . Both the groups showed initial loss of small myelinated fibres . No proliferation of Schwann cells was in FPI nerves, but in combined insult it was maximum after two weeks . Presence of M . leprae seems to be arresting Schwann cell activity after two weeks . Blood vessels showed increased endothelial cell cytoplasm, basement membrane proliferation and villi formation . These changes seem to be specific of endoneurial blood vessels of leprosy nerves . Increased number of mast cells seems to be specific of devascularized and FPI nerves . Increased number of macrophages expressed low immunity of devascularized nerves . Eosinophils migrated to endoneurium as a result of leakage of axoplasm. Age Ageing, 1992 Jan, 21(1), 5 - 12 Bacterial contamination of the small bowel in elderly people: is it necessarily pathological? Lipski PS, Kelly PJ, James OF. Bacterial contamination of the small bowel is probably the commonest cause of occult malabsorption in the elderly . It may occur in patients without a 'blind loop' or suggestive symptoms of diarrhoea and weight loss . We have prospectively studied the apparent prevalence of presumed bacterial contamination of the small bowel and its effect on nutritional state . Subjects were divided into three groups: (A) 54 young fit subjects; (B) 103 fit community elderly subjects; (C) 73 elderly long-stay hospital patients . All subjects had simultaneous lactulose hydrogen breath test and 14C-glycocholic acid breath test . Nutritional state was assessed by anthropometry, haematology and biochemistry . There were significantly fewer positive 14C-glycocholic acid breath tests in the young than in the elderly subjects (3% vs . 20% and 17%, p less than 0.0001) but no difference in the number of positive hydrogen breath tests between groups . There was no association between positive breath tests and anthropometry, haematology and biochemistry except for a lower albumin in group B and a lower red blood cell folate in group C with positive breath tests . These abnormal breath tests indicate that bacterial contamination of the small bowel may be common in normal fit elderly people and in elderly long-stay hospital patients and may be a concomitant of 'normal' ageing, not necessarily leading to ill-health. Cytometry, 1992, 13(2), 155 - 62 In vivo measure of average bacterial cell size from a polarized light scattering function; Bronk BV et al.; A particular combination of elements of the Mueller matrix for scattering of polarized light given by (S34 + S14)/(S11 + S13) identical to (S34/S11)++ is measured vs angle at a wavelength of 633 nm for randomly oriented suspensions of several species of bacteria in different stages of growth . (This combination of elements is dominated in the present measurements by the behavior of the normalized S34 matrix element, as is indicated by the notation defined on the right side of the equation.) The resulting graph in each case shows an oscillating function of angle . This function is compressed toward smaller angles when the bacteria are in the exponential phase of growth in comparison with results for a suspension of the same bacteria in the stationary (starving-smaller cells) phase of growth . Microscopic measurements were made to determine, for each case, the average dimensions of the bacterial population . Graphs were then plotted of the peak positions from the Mueller matrix function plots vs either cell length or cell diameter . The function was shown to be strongly correlated with cell diameter under the conditions of this experiment and poorly correlated with cell length . The measurements were shown to have a sensitivity to changes in average diameter of about 20 nm. Br J Obstet Gynaecol, 1992 Jan, 99(1), 63 - 6 Diagnosis of bacterial vaginosis in a gynaecology clinic; Hay PE et al.; OBJECTIVE: To estimate the prevalence of bacterial vaginosis in women referred to a gynaecology clinic, and to compare two methods of diagnosing bacterial vaginosis . SETTING: Gynaecology Clinic at Northwick Park Hospital SUBJECTS: 114 women aged 16 to 65 referred consecutively to the gynaecology clinic of one consultant . MAIN OUTCOME MEASURES: Detection of bacterial vaginosis by standard compound criteria and by examination of a Gram stained smear of fluid from the posterior vaginal fornix . RESULTS: Bacterial vaginosis was detected by both the Gram stain and the compound criteria in 13 women . There was no correlation between the symptom of vaginal discharge and the diagnosis of bacterial vaginosis in this population, but the presence of discharge noted by the clinician was associated with bacterial vaginosis . CONCLUSIONS: The prevalence of bacterial vaginosis was 11% . The Gram stain provides a simple and inexpensive method for laboratory confirmation of bacterial vaginosis where facilities for using the compound criteria are not available. Mol Microbiol, 1992 Jan, 6(2), 165 - 70 Direct visualization of plasmid DNA in bacterial cells; Eliasson A et al.; The direct visualization of plasmid DNA inside Escherichia coli cells is demonstrated using phase-fluorescence microscopy of DAPI (4',6-diamidino-2-phenylindole)-stained bacteria . Small as well as large plasmids could be detected, both in minicells and in cells of larger size . For large plasmids, even single molecules appeared to be within the detection limit . The fluorescence generated from monomers of small plasmids was probably below this limit, and for these plasmids the observed signals may represent aggregates . The distribution of the fluorescence foci might reflect specific plasmid positioning during partition and/or replication. Mamm Genome, 1992, 2(2), 87 - 95 The murine MHC encodes a mammalian homolog of bacterial ribosomal protein S13; MacMurray AJ et al.; The mouse major histocompatibility complex (MHC) contains many genes in addition to the classical immune response genes . We have screened overlapping cosmid clones covering 170 kb of the H-2K region for genes expressed in embryonal carcinoma (EC) cells . The Ke-3 gene (Abe et al . 1988) found in this region was further studied by Southern, Northern, and sequence analysis . It is an expressed, intron-containing locus encoding a mouse homolog of the bacterial ribosomal protein S13 . This is the first non-organelle S13 homolog identified in metazoans, and its genomic location has been determined precisely. Appl Environ Microbiol, 1992 Jan, 58(1), 111 - 8 Changes in bacterial composition and enzymatic activity in ileostomy and ileal reservoir during intermittent occlusion: a study using dogs; Ruseler-van Embden JG et al.; Bacterial flora, activities of 10 potential mucus- and dietary polysaccharide-degrading enzymes, blood group antigenicity of the intestinal glycoproteins, and proteolytic activity in the output from experimentally colectomized dogs with conventional ileostomies and dogs with valveless ileal reservoirs (pouches) were determined . The ileostomies of dogs with conventional surgery (group II) and with pouches (group III) were occluded intermittently during a 6-week period . The duration of occlusion was progressively increased . Group I, five dogs with conventional ileostomies, served as a control group . After occlusion of the ileal pouch for 7 h, total numbers of bacteria increased threefold, glycosidase activity increased fivefold, and blood group antigenicity of the intestinal glycoproteins, which was high in the output from the nonoccluded pouch, was no longer detectable . Proteolytic activity was not influenced by occlusion of the pouch . Significantly lower numbers of bacteria, only minor glycosidase activity, high blood group antigenicities of the intestinal glycoproteins, and higher proteolytic activity were found in ileostomy effluents from groups I and II . Histopathological examination showed chronic inflammation and changes in crypt-villus ratio in all dogs with ileal reservoirs; the ileal mucosa from the dogs with conventional ileostomies did not show any abnormalities . Consequences of the flora-related enzyme activities for the ileal mucosa are discussed. Scand J Thorac Cardiovasc Surg, 1992, 26(1), 79 - 82 Bacterial sedimentation during cardiac surgery reduced by disposable clothing; Lippert S et al.; The effect of three types of operating-room clothing on bacterial sedimentation was studied during cardiac valve surgery . The 24 operations were divided into three groups, according to type of clothing worn by all personnel--standard cotton, open style non-woven or closed style non-woven . Sedimentation on open agar plates was greatest when cotton clothing was worn, less with open style non-woven garments (difference not significant), and still less with closed style non-woven clothing (difference statistically significant) . The sedimentation corresponded to estimated airborne concentrations (cfu/m3) of 79.6 (cotton clothing), 59.7 (open style non-woven) and 7.2 (closed style non-woven) . Use of closed style non-woven operation clothing is recommended for cardiac surgery. Infection, 1992, 20 Suppl 1, S54 - 7 Cefodizime stimulates subpopulations of cells mediating spontaneous or antibody-dependent cytotoxicity in patients with bacterial infections; Auer IO et al.; The influence of cefodizime (CDZ) on spontaneous cell-mediated cytotoxicity (SCMC) and antibody-dependent cell-mediated cytotoxicity (ADCC) was investigated in nine patients with conditions predisposing to infection (T-cell deficiency, humoral immune deficiency, myeloproliferative syndrome, kidney or liver damage, or chronic pulmonary disease) . SCMC, using the K562 and LIK cell lines as targets, and ADCC, using the LIK cell line, were measured at baseline and after ten days of therapy with CDZ for lower respiratory tract infections . Six patients were cured; clinical outcome was not evaluable in the other three . SCMC lysis of K562 cells did not change significantly . SCMC and ADCC lysis of LIK (lymphoblastoid) cells both increased significantly . CDZ selectively stimulated a subpopulation of NK cells in this population of immunocompromised patients with lower respiratory tract infections. Scand J Immunol Suppl, 1992, 11, 91 - 5 Serodiagnosis of bacterial diseases: problems and developments; Terpstra WJ; Serodiagnosis is useful when pathogens cannot be detected quickly or easily by conventional methods or have been eliminated by the patient . Taking the example of leptospirosis, the microscopic agglutination test to detect specific antibodies after leptospires have disappeared from the patient's bloodstream is of unsurpassed reliability . Unfortunately the test is used mainly in specialized laboratories that can culture leptospires to serve as live antigens . Freeze-dried leptospires may allow the application of the test on a wide scale . The ELISA is useful for the detection of IgM antibodies in humans as a sign of current or recent infection . ELISA results must be confirmed by the microscopic agglutination test . A simplified easy-to-read ELISA using broadly reactive antigen for the detection of all types of leptospires might be useful for a quick detection of acute human leptospirosis . Using an appropriate antigenic preparation the ELISA is useful for the detection of antibodies in animals, e.g . cattle . As for pathogen detection, DNA-based methods such as PCR are promising for an early, quick and specific diagnosis . Antigenic analysis by monoclonal antibodies and genetic analysis by restriction fragment length polymorphism are for the time being complementary methods for the characterization of leptospiral isolates. Folia Microbiol (Praha), 1992, 37(2), 153 - 6 Functional and metabolic changes of guinea pig peritoneal macrophages induced by bacterial vaccines; Takacova J et al.; Phagocytic activity of peritoneal macrophages of guinea pigs injected subcutaneously with bacterial vaccines was found to be increased . The phagocytic index remained unchanged or was decreased . In addition, a decreased candidacidal activity was observed . Metabolic activation of macrophages, measured by the INT test, was inhibited in unstimulated cells while cells stimulated with zymosan or opsonized zymosan exhibited higher values of the INT test as compared with control animals . After injection of vaccines the number of peritoneal macrophages was increased, but the spleen mass decreased. Scand J Gastroenterol, 1992, 27(3), 253 - 6 Diagnosis of bacterial overgrowth after culturing proximal small-bowel aspirate obtained during routine upper gastrointestinal endoscopy; Bardhan PK et al.; The purpose of this study was to evaluate the method of obtaining aspirated fluid for culture from the small intestine through a fiberoptic gastrointestinal endoscope for diagnosing small-bowel overgrowth . The study population consisted of 10 healthy volunteers and 26 patients with various gastrointestinal problems referred for routine endoscopic examination . The material to be cultured was obtained under direct visualization approximately 25 to 30 cm distal to the pylorus or from the afferent loop (in Billroth-II patients) with a sterilized sheathed wash pipe passed through the suction channel of the endoscope . Cultures were considered positive for bacterial overgrowth if total counts of organisms were 10(5)/ml or more . All healthy volunteers and 16 of 21 unoperated patients had sterile or insignificant growth, whereas all 5 patients who had Billroth-II operations had positive overgrowth . The endoscopic method for collection of proximal gastrointestinal fluid for culture is simple and can be performed during routine endoscopy. Zh Mikrobiol Epidemiol Immunobiol, 1992, (9-10), 2 - 5 {Associations of the soil amoeba Hartmannella rhysodes with the bacterial causative agents of plague and pseudotuberculosis in an experiment}; Nikul'shin SV et al.; Some aspects of relationships between soil ameba and the causative agents of plague and pseudotuberculosis, capable of forming natural associations, are considered . Ameba can phagocytose bacteria causing plague and pseudotuberculosis . Cases of the preservation of individual bacterial cells at the stationary phase and in precysts of amebae have been registered. Scand J Rheumatol, 1992, 21(6), 283 - 8 Synovial fluid leukocytosis in bacterial arthritis vs . reactive arthritis and rheumatoid arthritis in the adult knee; Kortekangas P et al.; In this comparative analysis of laboratory data, we examined the characteristics of synovial fluid leukocytosis in eighty adult patients with bacterial arthritis, reactive arthritis or rheumatoid arthritis of the knee joint . Synovial fluid leukocyte count and the percentage of polymorphonuclear cells seemed to perform well as a discriminator between bacterial infection and acute flare of the underlying disease in patients with rheumatoid arthritis . In contrast, there were no definite difference in the intensity of synovial fluid leukocytosis between patients with bacterial arthritis caused by living bacteria and patients with reactive arthritis probably caused by bacterial antigens. Vaccine, 1992, 10(14), 977 - 90 Bacterial vaccines: old and new, veterinary and medical; Walker PD; Developments in veterinary and medical bacterial vaccines are outlined . In the former case, economic considerations are paramount, and cruder, less purified products of proven efficacy continue to be employed . For human use, however, safety and absence of side effects are increasingly demanded . Various examples of human and veterinary vaccines are discussed, and interaction between the two fields is illustrated by reference to the pig-bel disease in New Guinea and the possible aetiology of Sudden Infant Death Syndrome. HPB Surg, 1992, 6(1), 23 - 32; discussion 33 Primary bile duct stones and bacterial activity; Vitetta L et al.; The results of this study suggest that infection with beta-glucuronidase active bacteria is the initial event in the nucleation of primary bile duct stones (PBDS) . PBDS from five patients were morphologically fragile and "earthy" with alternating light and dark brown pigment layers with no evidence of a distinct central nucleus that may have been reminiscent of a different structure . Chemically, calcium bilirubinate and calcium palmitate were prominent throughout their structure . All bile duct biles had a positive culture and were always associated with at least one bacterial species which was beta-glucuronidase active . Moreover, fragments of PBDS nuclear areas had positive cultures that were comparable with those present in their individual bile duct bile . Microscopic examination of bile showed abundant precipitation of calcium bilirubinate granules in all samples . Thus, bile duct bile infection with beta-glucuronidase active bacteria (e.g . E . coli, C . perfringens) appears to be a key factor in PBDS pathogenesis, having a precursor role, rather than being a consequence . Bile stasis is likely to be a co-factor which must have a supportive role in subsequent stone growth. Acta Physiol Scand Suppl, 1992, 607, 193 - 9 Kdp, a bacterial P-type ATPase whose expression and activity are regulated by turgor pressure; Epstein W; The Kdp ATPase is a P-type ATPase consisting of three large protein subunits in a complex that probably contains 2 copies of each subunit . A small hydrophobic peptide, encoded in the same operon as the large subunits, may also participate . Kdp has very high affinity for K+ and serves to scavenge this ion when its concentration is low . Kdp responds to turgor pressure at two levels, at the level of activity and of expression . Kdp mediates net uptake when turgor is low, but mediates exchange without net change when turgor is normal . Kdp is expressed only when turgor is low . This control is mediated by a pair of regulatory proteins, members of the class of 'sensor-effector' regulators widely distributed in bacteria . It is suggested that low turgor changes the conformation of the KdpD 'sensor' protein, activating its presumed kinase activity to phosphorylate KdpE, the 'effector' protein, and phospho-KdpE in turn turns on expression of the operon that encodes the Kdp complex. Annu Rev Microbiol, 1992, 46, 277 - 305 The electron-transport proteins of hydroxylating bacterial dioxygenases; Mason JR et al.; The degradation of aromatic compounds by aerobic bacteria frequently begins with the dihydroxylation of the substrate by nonheme iron-containing dioxygenases . These enzymes consist of two or three soluble proteins that interact to form an electron-transport chain that transfers electrons from reduced nucleotides (NADH) via flavin and {2Fe-2S} redox centers to a terminal dioxygenase . The dioxygenases may be classified in terms of the number of constituent components and the nature of the redox centers . Class I consists of two-component enzymes in which the first protein is a reductase containing both a flavin and a {2Fe-2S} redox center and the second component is the oxygenase; Class II consists of three-component enzymes in which the flavin and {2Fe-2S} redox centers of the reductase are on a separate flavoprotein and ferredoxin, respectively; and Class III consists of three-component enzymes in which the reductase contains both a flavin and {2Fe-2S} redox center but also requires a second {2Fe-2S} center on a ferredoxin for electron transfer to the terminal oxygenase . Further subdivision is based on the the type of flavin (FMN or FAD) in the reductase, the coordination of the {2Fe-2S} center in the ferredoxin, and the number of terminal oxygenase subunits . From the deduced amino acid sequence of several dioxygenases the ligands involved in the coordination of the nucleotides, iron-sulfur centers, and mononuclear nonheme iron active site are proposed . On the basis of their spectroscopic properties and unusually high redox potentials, the {2Fe-2S} clusters of the ferredoxins and terminal oxygenases have been assigned to the class of Rieske-type iron-sulfur proteins . The iron atoms in the Rieske iron-sulfur cluster are coordinated to the protein by two histidine nitrogens and two cysteine sulfurs. Agents Actions, 1992, Spec No, C112 - 4 Desferrioxamine potentiated SOD antiinflammatory activity in rat adjuvant arthritis: role of iron and bacterial toxins; Benoni G et al.; In this paper we studied the modulating inflammatory activity of iron in the adjuvant arthritis, taking indomethacin as a standard antiinflammatory drug and a superoxide dismutase derivative (MPEG-SOD) as a scavenger of free radicals . Moreover, we evaluated the changes in potential intestinal pathogens requiring iron for growth, in order to study the role of bacteria in the altered gastrointestinal functions observed during arthritis . We observed a 50% arthritis inhibition on the 14th day with MPEG-SOD plus desferrioxamine, a significant decrease in serum iron in arthritic rats compared to controls, and a significant Cl . perfringens increase on the 28th day in the presence of MPEG-SOD . Our data demonstrate that hypoferremia, in arthritis, is a protective mechanism overall in the early phase and could protect the intestinal tract by inhibiting the development of potential pathogens. Acta Otolaryngol, 1992, 112(3), 530 - 8 Nasopharyngeal bacterial flora in otitis prone children treated with immunoglobulin; Jorgensen F et al.; The present study was undertaken to evaluate possible beneficial effects of regularly given, long term immunoglobulin prophylaxis of children below 2 years of age with recurrent acute otitis media (RAOM) . The nasopharyngeal bacterial flora and the frequency of acute otitis media (AOM) and secretory otitis media SOM were studied . Every second of 44 children with 3 or more periods of AOM during the last year received immunoglobulin intramuscularly (Gammaglobulin Kabi 165 mg/l, 0.45 ml/kg b.w.) every third week for 6 months, while the other 22 children served as controls . All children were followed for 12 months . Immunoglobulin prophylaxis neither influenced the nasopharyngeal flora, nor the frequency of AOM or SOM periods . Children with AOM or SOM more often harbored bacterial pathogens in their nasopharynx than children with normal middle ear status . Also, the immunoglobulin prophylaxis did not influence the increased frequency of bacterial pathogens in the nasopharynx of children attending public day care or family day care as compared to those taken care of at home. Nutr Cancer, 1992, 18(2), 157 - 64 Bile salt/acid induction of DNA damage in bacterial cells: effect of taurine conjugation; Zheng ZY et al.; Bile salts and acids have been implicated in the etiology of colon cancer, possibly through their ability to cause DNA damage . Taurine-conjugated and nonconjugated forms of three bile salts and one bile acid were tested for DNA repair-inducing potential and for cellular toxicity in a recently developed Escherichia coli chromotest system . The taurine-conjugated forms of sodium deoxycholate and lithocholic acid had reduced ability to induce DNA repair . Also the taurine-conjugated form of lithocholic acid had a reduced lethal effect . These observations suggest that the biotransformation step, whereby bacteria in the intestine remove the taurine added to bile salts in the liver, may be significant in the etiology of colon cancer. Dev Biol Stand, 1992, 77, 49 - 56 Synthetic lipopeptide immunomodulators derived from bacterial lipoprotein: tools for the standardization of in vitro assays; Bessler WG; For the evaluation of immunomodulators by in vitro assays, agents working reproducibly are difficult to obtain; conventional preparations of bacterial immunomodulators tend to vary for different preparations . Here we suggest that synthetic lipopeptide analogues derived from bacterial lipoprotein can be used as standards for various in vitro assays: studying B lymphocyte activation, lipopeptides act as potent mitogens and polyclonal activators inducing immunoglobulin synthesis . In monocytes/macrophages, lipopeptide stimulate the secretion of IL-1, IL-6, tumor necrosis factor (TNF) and nitrogen oxide (NO); they also induce tumor cytotoxicity . Lipopeptides also constitute potent immuno-adjuvants in vitro and in vivo, either in combination with or covalently bound to antigen . These activities are displayed in various species; in mice they are found in LPS non-responder and responder strains . The novel synthetic lipopeptides described here can be synthesized readily in gram amounts with high purity and reproducibility; they are non-toxic and can be stored for a long time even at room temperature . Thus, lipopeptides meet the requirements to serve as effective standards for a multitude of relevant biological assays. J Virol Methods, 1992 Jan, 36(1), 35 - 49 Use of bacterial trpE fusion vectors to express and characterize the bovine immunodeficiency-like virus core protein; Atkinson B et al.; The gag coding region from Bovine Immunodeficiency-like Virus (BIV) was cloned into E . coli and expressed as a bacterial fusion protein . Six different clones spanning various regions of the gag open reading frame were generated . The resulting fusion proteins were expressed at high concentrations and readily purified . A panel of bovine immune sera specifically recognized the recombinant Gag proteins, as did immune sera from animals infected or immunized with lentiviruses related to BIV, such as Equine Infectious Anemia Virus (EIAV) and Human Immunodeficiency Virus (HIV) . Analysis of the deletion clones, using the bovine immune sera panel, enabled us to identify at least one major epitope which was specifically recognized by all bovine sera examined . The ease of expression, purification, and specificity of these fusion proteins should enable a thorough study of the epidemiology of BIV infection. J Leukoc Biol, 1992 Jan, 51(1), 93 - 6 Synergistic interaction of bacterial lipopolysaccharide and the monocyte-macrophage colony-stimulating factor: potential quantitative and qualitative changes in macrophage-produced cytokine bioactivity; Evans R et al.; In this brief definitive report, we show that over a 6-h period and under serum-free conditions, recombinant monocyte-macrophage colony-stimulating factor 1 (rCSF-1) and lipopolysaccharide (LPS) synergize and induce macrophages to express higher levels of mRNA for interleukin 1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor alpha (TNF-alpha), and IL-6 and to release more bioactivity than macrophages treated with LPS alone . This synergy was regulated by the amount of LPS in the culture medium . Paraformaldehyde-fixed macrophages like-wise showed augmentation of IL-1 activity, but whereas all of the bioactivity associated with the fixed macrophages could be neutralized by anti-IL-1 alpha antibody only approximately 40% of the supernate activity could be attributed to IL-1 alpha . Preliminary data suggest that the augmenting effect induced by CSF-1 cannot be explained solely on a quantitative basis because the addition of rIL-1 alpha to supernates of macrophages treated with LPS alone or with the combination of LPS and CSF-1 resulted in an increase in thymocyte mitogenic activity to a level that could not be explained on an additive basis . Although the supernates contained TNF and IL-6, antibody neutralization assays made it unlikely that these were directly responsible for the augmenting effect . These results suggest that CSF-1 not only enhances basic genetic responses induced by LPS alone but also may induce a mechanism that amplifies cytokine bioactivity. Teratog Carcinog Mutagen, 1992, 12(3), 97 - 112 Further in vitro and in vivo mutagenicity assays with thiram and ziram fungicides: bacterial reversion assays and mouse micronucleus test; Crebelli R et al.; The fungicides thiram and ziram have been assayed in a battery of nine bacterial strains of different genetic specificity . The results obtained suggest the induction of excisable DNA lesion(s), and indicate similar mutability of strains with AT or GC base pairs at target sites . This mutagenic profile is clearly distinct from that of oxidative mutagens, and it does not support the proposed role of oxidative stress in the mechanism of dithiocarbamates mutagenicity in bacteria . Furthermore, the bone marrow micronucleus test has been carried out in B6C3F1 mice with intraperitoneal administration of high grade thiram (12.5-50 mg/kg) and ziram samples (2.5-10 mg/kg in males, and 5-20 mg/kg in females) . Thiram produced a significant increase of micronucleated PCEs in male mice sampled 48 h after treatment with 25, 37.5, and 50 mg/kg . No significant increase was detected in treated females . Ziram, tested in a lower range of doses because of its higher toxicity, resulted negative in both sexes . Both the acute toxicity and the ratio polychromatic/normochromatic erythrocytes indicated some sex specificity in the toxic effects induced by these dithiocarbamates in the B6C3F1 mouse. Adv Exp Med Biol, 1992, 319, 31 - 8 Bacterial polysaccharides, endotoxins, and immunomodulation; Baker PJ et al.; These studies show that at least some--though certainly not all--of the adjuvant effects of LPS and its derivatives can be attributed to its ability to eliminate the inhibitory effects of Ts which are activated during the course of a normal immune response . The ability of nontoxic MPL to act in this fashion suggests that it can be used as a safe and acceptable alternative to Freund's complete adjuvant to increase the immunogenicity of poorly immunogenic antigens . More important, the ability of MPL to eliminate the expression of Ts activity, without adversely influencing other T cell functions (e.g., Th, Ta, and Tc activity) makes its use as an adjuvant even more promising since it can then permit those T cell functions to be expressed in a much more efficient manner . Obviously, this would have great significance for the development of tumor immunity. Acta Physiol Hung, 1992, 79(4), 355 - 63 CRF-dependent and CRF-independent mechanisms involved in hypophysial-adrenal system activation by bacterial endotoxin; Elenkov IJ et al.; The immune system and the hypothalamic-pituitary-adrenal (HPA) axis play important role in the overall inflammatory response . The mechanism through which lipopolysaccharide (LPS, endotoxin) stimulates the HPA axis is not well understood . In order to clarify the role of hypophysiotropic peptides of paraventricular origin in the effect of LPS on ACTH and corticosterone secretion, the effect of LPS was studied on rats with lesions of hypothalamic paraventricular nucleus (PVN) . It was shown that 90 min after 2 mg/kg LPS i.p . the ACTH, but not the corticosterone response was effectively blunted in PVN-lesioned rats, as compared to sham operated animals . However, in PVN-lesioned rats 240 min after treatment with LPS a significantly higher plasma ACTH and corticosterone level was monitored . It is, therefore, suggested that in response to LPS activation of HPA both CRF(s)-dependent and CRF(s)-independent mechanisms are involved, even a direct effect of the adrenal cortex should be taken into account. Acta Neurochir (Wien), 1992, 119(1-4), 121 - 7 The influence of bacterial collagenase on regeneration of severed rat sciatic nerves; Wehling P et al.; Regeneration of peripheral nerve fibers is impeded by the formation of scar tissue at the site of injury . The possible beneficial effect of collagenase on nerve regeneration was studied using clinical, neurophysiological (evoked potentials) and histological (nerve fiber counts) methods . The sciatic nerves of rats were transected and the severed ends abutted and sewn together . In one series, the area about the lesion was covered with fibrin adhesive and infused with either isotonic saline (controls) or collagenase (treatment group) . In the other series, the severed ends of the nerve were inserted into a silicone tube and separated by a collagen plug, which was infused with either saline or collagenase . Compared to the controls, the treated animals showed a significant improvement of clinical and neurophysiological parameters . After 3 months of observation, the collagen content of the transection site was reduced, and in the silicone series, the total number of myelinated axons 5 mm distal to the site of transection was increased, while the fiber diameter distribution was unchanged. Zh Mikrobiol Epidemiol Immunobiol, 1992, (5-6), 25 - 7 {The rapid differential diagnosis of bacterial and viral meningitis by using the lysozyme test}; Babich EM et al.; The authors have modified the technique of the lysozyme test by adding polimixin M sulfate into the gel bacterial medium . Rapid diagnosis with the use of this test is based on different time of the appearance of the lysis areas: in bacterial meningitides the CSF lysozyme activity is detectable within 15-120 min, whereas in viral meningitides it manifests 40-50 min later or does not manifest at all . The results were found to depend on the time of the CSF collection: the earlier the CSF samples were obtained, the higher was the share of positive results. Zh Nevropatol Psikhiatr Im S S Korsakova, 1992, 92(2), 36 - 8 {Permeability of the blood-brain barrier for metals in bacterial meningoencephalitis}; Iarosh OA; A study was made of blood-brain barrier permeability for metals in 50 patients depending on the period and gravity of bacterial meningoencephalitis . Selective permeability of the blood-brain barrier for metals is aimed in different periods of the inflammatory process at the retention of the mineral composition of the cerebrospinal fluid within the limits required for normal function of the central nervous system. Zh Nevropatol Psikhiatr Im S S Korsakova, 1992, 92(2), 33 - 6 {Mathematical analysis of permeability of the blood-brain barrier in bacterial meningoencephalitis}; Iarosh OA; Atomic-absorption spectrophotometry was used to measure the concentration of certain metals in the blood and CSF in 70 patients suffering from bacterial meningoencephalitis depending on the gravity and stage of the inflammatory process . Probability statistical and correlation analysis of the content of metals in the blood and CSF made in possible to depict the status of permeability and different transformations that regulate the functions of the blood-brain barrier in patients seen over time. Zh Nevropatol Psikhiatr Im S S Korsakova, 1992, 92(1), 48 - 53 {Clinico-morphological characteristics of cardiogenic non-bacterial cerebral embolism}; Shmushkevich LS et al.; Clinical and morphological studies were carried out in 194 patients who died in the acute period of ischemic brain stroke . Disorders of the heart rhythm were found to play the leading part in the origin of cardiogenic embolisms of cerebral vessels . It has been recorded that left atrial thrombus formation was prevalent in heart rhythm disorders whereas left ventricular was prevalent in lack of those disorders . In different heart diseases complicated by atrial fibrillation and cerebral thromboembolism, the one-third of cases did not show any heart thromboses . The content of embolisms in cerebral arteries corresponded to the material of their source, whereas the site was specified by mechanical obstruction, with no changes in the vascular wall . It has been disclosed that in the majority of cases, stable embolism precipitates white infarction or white infarction with the hemorrhagic component whereas embolus lysis is responsible for hemorrhagic infarction and (or) a transitory ischemic attack . The clinical manifestations of hemorrhagic infarction differ from white infarction in less gravity and regression of the neurological symptomatology. Endocr Res, 1992, 18(1), 31 - 40 Cyclic guanosine monophosphate analogs do not reverse bacterial toxin modulation of lactogen-stimulated NB2 cell mitogenesis; Larsen JL; Pertussis toxin (PT) and cholera toxin (CT) have been shown to modulate lactogenic hormone-stimulated Nb2 cell mitogenesis, a lactogen-dependent cell line . As both toxins have been shown to alter guanylate cyclase activity in other cell systems, cyclic guanosine monophosphate (cGMP) analogs, 8-bromo or dibutyryl cGMP, were added to determine if they could reverse the toxin-mediated effects . In the absence of bacterial toxins, both cGMP analogs enhanced lactogen-stimulated Nb2 cell mitogenesis in a multiphasic pattern . At maximal enhancement, the effect was statistically significant but not marked (113 +/- 5%; p less than 0.01) . Neither cGMP analog increased lactogenic binding site number or affinity so cGMP must affect lactogen action following receptor binding . Neither analog could stimulation Nb2 cell mitogenesis in the absence of lactogens so cGMP is not a second messenger for lactogens in this cell system . Finally, neither cGMP analog reversed the inhibitory effects of either bacterial toxin on lactogen-stimulated Nb2 cell proliferation . In summary, although bacterial toxins may be capable of altering guanylate cyclase activity, as addition of cGMP analogs do not reverse toxin-mediated effects on lactogen-stimulated mitogenesis, these toxins' actions must be mediated predominantly through other mechanisms that may have significant importance to lactogen signal transduction. Eur J Cardiothorac Surg, 1992, 6(5), 272 - 3 Surgical treatment of persistent ductus arteriosus complicated by bacterial endocarditis; Stejskal L et al.; A seven-year-old girl with persistent ductus arteriosus developed acute bacterial endocarditis with large vegetations in the ductus and the left pulmonary artery . Conservative management was not successful: repeated embolisation to the lungs necessitated operation during the acute endocarditis . Details of the operative technique are presented and discussed. Nauchnye Doki Vyss Shkoly Biol Nauki, 1992, (4), 87 - 9 {The antiviral action of a modified bacterial ribonuclease}; Indulen MK et al.; The antiviral activity of a bacterial ribonuclease conjugate with chitosane of Kamchatka crab (in a form of water soluble chito-oligosaccharides) has been studied . The conjugate inhibitory activity for A and B viruses as well as to Sindbis arbovirus in tissue cultures is shown . The preparation efficiency at intramuscular and intranasal administration was observed at experimental influenza infection of white mice. Nauchnye Doki Vyss Shkoly Biol Nauki, 1992, (2), 71 - 7 {The heat denaturation of bacterial ribonuclease studied by infrared spectroscopy}; Strupishina EA; The thermal denaturation of bacterial ribonuclease in the interval of pH 2.5-7.0 has been investigated by means of infra-red spectroscopy method . The protein melting for pH 2.5 begins at the temperature 25 degrees C and is accompanied by secondary protein structure reconstruction, partially destroying native beta-structure and leading to new denatured conformation appearance of different types of beta-turns . Spectral changes for pH 3.5 and 7.0 are significantly less in the same frequency areas . At the temperature more than 50 degrees C protein aggregation takes place with inter-molecule-beta-form formation. Zh Mikrobiol Epidemiol Immunobiol, 1992, (11-12), 43 - 6 {The use of solid-phase liposomal immunoassay for determining a bacterial antigen of lipopolysaccharide nature}; Tret'iakov SI et al.; The solid-phase liposomal immunoassay procedure for the determination of Francisella tularensis lipopolysaccharide (LPS) has been developed . This assay has been made with the use of monolayer liposomes, on the average, 360 nm in diameter with their phospholipid bilayer modified with F . tularensis LPS and their internal space filled with calcein used as fluorescent marker . The assay is based on the principle of the competitive immunosorption of liposomes and antigenic LPS on the surface of polystyrene plates sensitized with specific monoclonal antibodies . The dynamic range of the method is 50-2,500 mg/ml, the variation index being 2.3-11.5% . Optimization of this method and its comparison with the enzyme immunoassay system for the given antigen-antibody pair has been made. Acta Med Austriaca, 1992, 19(5), 133 - 6 {The value of echocardiography in diagnosis of bacterial endocarditis}; Spinka R et al.; In recent years echocardiography has become the most important method for the diagnosis of bacterial endocarditis . Though identification of valve vegetations by M-mode echocardiography was rather limited, two dimensional echocardiography--especially with transoesophageal application--has a sensitivity of more than 90% and a specificity of 98% . In addition, echocardiography is able to supply valuable contributions for the evaluation of the patient's prognosis and in the identification of complications of infective endocarditis, such as: the extent of valve destruction, rupture of chordeae and other parts of the valves, as well as the presence of intramyocardial and paravalvular abscesses . In addition, the application of Doppler-echocardiography allows the semiquantitative evaluation of the degree of any valve insufficiency. Med Dosw Mikrobiol, 1992, 44(1-2), 29 - 34 {The role of bacterial metabolism in transformation of non-mutagenic compounds into mutagens . I . Participation of Escherichia coli nitroreductase in creation of mutagens from non-mutagenic new pesticides}; Szarapinska-Kwaszewska J et al.; Investigations concerned Escherichia coli nitroreductase in creation of mutagens from non-mutagenic pesticides-derivatives of urea . Three new compounds were studied: N-phenyl-N'-methylurea (IPO 4328), N-methyl,N-(2-hydroxyethyl)-N'phenylurea (IPO 2363), N-(2-hydroxyethyl), N-methyl-N'-(3,4 dichloroethyl) urea, and diurone-3-(3,4 dichlorophenyl)-1,1 dimethylurea . These compounds were incubated in anaerobic conditions with cells of E . coli K-12 (KF) strain and nitrate or nitrite . Using Ames test, mutagenicity of resulting metabolites was investigated . It was found that during incubation of herbicide IPO 4328 with cells of E . coli K-12 (KF) and nitrate, mutagenic product for strain of S . typhimurium TA 1537 is created . Very weak mutagenic metabolite for the same strain was appearing during incubation of herbicide IPO 2363 with cells of E . coli K-12 (KF) in presence of nitrite . Incubation of investigated compounds with E . coli K-12 (KF) cells alone did not result in appearance of mutagenic substances . Thus, role of Escherichia coli in creation of mutagenic compounds from non-mutagenic derivatives of urea consisted of nitrite from nitrate production with participation of nitroreductase, which afterwards in absence of bacteria or action of their enzymes reacted with investigated pesticides. Invasion Metastasis, 1992, 12(3-4), 197 - 209 Altered establishment/clearance mechanisms during experimental micrometastasis with live and/or disabled bacterial lacZ-tagged tumor cells; Lin WC et al.; To study micrometastasis at its earliest stages, the bacterial lacZ marker gene was introduced into human EJ Ha-ras-transformed BALB/c 3T3 cells (LZEJ), followed by their intravenous injection into nude mice . Lung micrometastases were easily identified by blue staining of lacZ-tagged cells minutes/hours after injection, permitting effective evaluation of establishment/clearance mechanisms of LZEJ cells . Different treatments were used to disable LZEJ cells (fixation, irradiation, or mitomycin C) to determine modulation of these processes--although unable to divide, these cells stain for lacZ expression for days after treatment . Fixation-killed cells generated large microfoci (> 13-15 cells/focus) with well-rounded morphologies while live, irradiated, or mitomycin-treated cells generated smaller, irregularly shaped foci (3-7 cells/focus) . Fixed-cell foci were cleared more slowly from lungs than the other three classes, even when prefiltered to remove large aggregates . All foci of disabled cells were eventually cleared while a basal level of live-cell foci persisted . Co-injection of fixed and live cells (or preinjection of fixed cells, followed by live cells) resulted in complete clearance of live-cell microfoci; in contrast, preinjection of live cells (then injection of fixed cells) led to survival of live-cell micrometastases . Therefore, altered deformability and/or cell surface interactions of tumor cells modulate the effectiveness of host-clearing mechanisms in the lung and in some situations these altered cells facilitate clearance of live tumor cells that are normally tumor-progressing. Ann Biol Clin (Paris), 1992, 50(8), 587 - 91 Evaluation of bronchoalveolar lavage for the diagnosis of bacterial pneumonia in ventilated patients; Roger-Moreau I et al.; Fourty-six patients requiring mechanical ventilation and suspected of bacterial pneumonia were examined by fiberoptic bronchoscopy . Specimens recovered by bronchoalveolar lavage (BAL) and using a protected specimen brush (PSB) were quantitatively cultured and the results compared . An assessment of the percentage of cells with intracellular organisms present on cytocentrifuged preparations made from lavage fluid was made to evaluate the utility of this method in early diagnosis of pneumonia . BAL cultures made a correct diagnosis in 43 out of 46 patients and detected 10 false-negative cultures of the PSB . With a threshold of more than 3% of cells with intracellular bacteria, direct microscopic examination diagnosed bacterial pneumonia in 36 out of 46 patients and allowed appropriate antibiotherapy to be instituted earlier . These results demonstrate the ability of BAL to diagnose bacterial pneumonia in ventilated patients. Rocz Akad Med Bialymst, 1992, 37, 64 - 70 The serum immunoglobulins level in maxillofacial bacterial infections; Zolkowski K et al.; The IgG, IgA and IgM levels in the serum of 23 patients with maxillofacial bacterial infections were determined . The high IgG level noted on admission further increased in the serum of 14 patients with acute inflammation and decreased in the serum of 10 patients with a chronic course . The level of IgM in the serum of patients with acute inflammation was higher than in the patients with chronic inflammation and decreased in both groups on the 7th day of treatment . The IgA level was high on admission in both groups and fell on the 7th day of treatment . Our results suggest that analysis of the serum level of IgG, IgA and IgM may be of diagnostic and prognostic value in the treatment of maxillofacial bacterial inflammation. J Cardiovasc Pharmacol, 1992, 20 Suppl 12, S135 - 8 Endothelium-derived kinins account for the immediate response of endothelial cells to bacterial lipopolysaccharide; Fleming I et al.; We have recently shown that cultured endothelial cells produce kinins that can stimulate endothelial nitric oxide (NO) production in an autocrine manner . Because both the kallikrein-kinin system and the L-arginine/NO pathway have been implicated in the pathogenesis of septic shock, we investigated the possible involvement of endothelium-derived kinins in the response of cultured endothelial cells to bacterial lipopolysaccharide (LPS) . In primary cultures of human umbilical vein and porcine aortic endothelial cells, LPS (0.3 to 3 micrograms/ml) induced significant concentration-dependent increases in cyclic GMP and 6-keto-PGF1 alpha, both of which were abolished in the presence of the selective bradykinin B2-receptor antagonist HOE 140 (0.1 microM) . These LPS-induced increases in cyclic GMP and 6-keto-PGF1 alpha were short lived, being maximal after 5 min but were not apparent after 60 min . In parallel with these effects, LPS (30 micrograms/ml) induced a distinct, HOE 140-sensitive increase in the intracellular calcium concentration of human endothelial cells loaded with indo-1 . In summary, these data suggest that the release of endothelium-derived kinin and subsequent stimulation of endothelial cells, followed by the enhanced production of NO and prostacyclin (PGI2), are implicated in the immediate hypotension induced by LPS in vivo. Proc Natl Acad Sci U S A, 1991 Dec 15, 88(24), 11207 - 11 Rapid-flow resonance Raman spectroscopy of bacterial photosynthetic reaction centers; Shreve AP et al.; Rapid-flow resonance Raman vibrational spectra of bacterial photosynthetic reaction centers from the R-26 mutant of Rhodobacter sphaeroides have been obtained by using excitation wavelengths (810-910 nm) resonant with the lowest energy, photochemically active electronic absorption . The technique of shifted excitation Raman difference spectroscopy is used to identify genuine Raman scattering bands in the presence of a large fluorescence background . The comparison of spectra obtained from untreated reaction centers and from reaction centers treated with the oxidant K3Fe(CN)6 demonstrates that resonance enhancement is obtained from the special pair . Relatively strong Raman scattering is observed for special pair vibrations with frequencies of 36, 94, 127, 202, 730, and 898 cm-1; other modes are observed at 71, 337, and 685 cm-1 . Qualitative Raman excitation profiles are reported for some of the strong modes, and resonance enhancement is observed to occur throughout the near-IR absorption band of the special pair . These Raman data determine which vibrations are coupled to the optical absorption in the special pair and, thus, probe the nuclear motion that occurs after electronic excitation . Implications for the interpretation of previous hole-burning experiments and for the excited-state dynamics and photochemistry of reaction centers are discussed. Proc Natl Acad Sci U S A, 1991 Dec 15, 88(24), 11202 - 6 Mechanism of the initial charge separation in bacterial photosynthetic reaction centers; Chan CK et al.; The initial electron transfer in reaction centers from Rhodobacter sphaeroides R26 was studied by a subpicosecond transient pump-probe technique . The measured kinetics at various wavelengths were analyzed and compared with several mechanisms for electron transfer . An unambiguous determination of the initial electron transfer mechanism in reaction centers cannot be made by studying the anion absorption region (640-690 nm), due to the spectral congestion in this region . However, correlations between the stimulated emission decay of the excited state of the special pair, P*, at 926 nm and bleaching of the bacteriopheophytin Qx absorption at 545 nm suggest that the electron transfer at 283 K is dominated by a two-step sequential mechanism, whereas one-step superexchange and the two-step sequential mechanism have about equal contributions at 22 K. Eur J Obstet Gynecol Reprod Biol, 1991 Dec 13, 42(3), 217 - 20 Bacterial vaginosis and the vaginal leucocyte/epithelial cell ratio in women attending an outpatient gynaecology clinic; Larsson PG et al.; In order to investigate the usefulness of vaginal leucocytosis as a sign of genital infection, vaginal wet smears were obtained from 230 consecutive non-pregnant women attending an outpatient gynaecology clinic . Although 52 (22.6%) of the women were diagnosed as having bacterial vaginosis, none had symptoms of cervicitis or genital infection . Of the patients with bacterial vaginosis 19 (36.5%) had vaginal leucocytosis compared to 37.1% of the women without bacterial vaginosis . In a further study, bacterial vaginosis was also diagnosed in 104 of 384 women (33.3%) attending for first trimester legal abortions . Of the women with bacterial vaginosis 37.5% had vaginal leucocytosis compared to 31.8% for those without bacterial vaginosis (P greater than 0.05) . There was no correlation between vaginal leucocytosis and the presence of cervical Chlamydia trachomatis; however, the presence of C . trachomatis did correlate with bacterial vaginosis . Vaginal leucocytes varies with factors unrelated to genital infection . This fact must be taken into consideration when wet smears from patients with suspected genital tract infection are evaluated. Science, 1991 Dec 13, 254(5038), 1627 - 30 Molecular architecture and electrostatic properties of a bacterial porin; Weiss MS et al.; The integral membrane protein porin from Rhodobacter capsulatus consists of three tightly associated 16-stranded beta barrels that give rise to three distinct diffusion channels for small solutes through the outer membrane . The x-ray structure of this porin has revealed details of its shape, the residue distributions within the pore and at the membrane-facing surface, and the location of calcium sites . The electrostatic potential has been calculated and related to function . Moreover, potential calculations were found to predict the Ca2+ sites. Protein Seq Data Anal, 1991 Dec, 4(6), 349 - 53 Bacterial cellulose-binding domain-like sequences in eucaryotic polypeptides; Meinke A et al.; The catalytic domain of endoglucanase II of Trichoderma reesei and the spore germination-specific polypeptide 270-11 of Dictyostelium discoideum contain amino acid sequences which share identity with the sequences of the cellulose-binding domains of several bacterial beta-1,4-glycanases . This is the first report of the presence of such sequences in eucaryotic polypeptides. J Gynecol Surg, 1991 Winter, 7(4), 233 - 7 Bacterial contamination at abdominal hysterectomy: a comparison of staple closure with regular suture closure of the vaginal wall; Beresford JM et al.; Bacterial contamination of the pelvis from the vagina at abdominal hysterectomy is assumed to be a major cause of infectious complications . A pilot study was completed to compare the bacterial contamination that occurred when the vaginal vault was closed using an automatic stapling technique with a regular suture method . The findings suggested that there was considerably less contamination using the staple technique and that infectious complications were associated with heavy bacterial contamination from the vagina . More extensive studies of this association are needed to confirm the presented data. J Mol Biol, 1991 Dec 5, 222(3), 465 - 77 Programmed translational frameshifting and initiation at an AUU codon in gene expression of bacterial insertion sequence IS911; Polard P et al.; The proteins expressed by insertion sequence IS911, a member of the widespread IS3 family of elements, have been analyzed . The results indicate that three major species are produced from two consecutive reading frames . A protein of Mr 11,500, ORFA, is synthesized from an upstream reading frame . A larger protein, ORFAB, uses the same initiation codon and is produced by a -1 programmed translational frameshift between orfA and a downstream frame, orfB, whose amino acid sequence shows significant homology with retroviral integrase proteins . The orfB frame is also expressed independently in two alternative forms: the first uses a rare AUU initiation codon in the orfB phase whereas the second appears to initiate in the orfA phase and is produced by a -1 frameshift mechanism similar to that used in ORFAB expression . A specific IS911 integration reaction using a minimal active junction composed of 51 base-pairs of the right inverted repeat and a flanking phase lambda sequence resembling a second end in inverted orientation has been developed to analyze the functions of these proteins by transcomplementation in vivo . The orfA and orfB frames are shown to be essential and production of ORFAB is shown to stimulate integration in this system, suggesting that this fusion protein is the IS911 transposase. Proc Natl Acad Sci U S A, 1991 Dec 1, 88(23), 10470 - 4 Characterization of the interaction of plant transcription factors using a bacterial repressor protein; Frohberg C et al.; Transcription initiation from a eukaryotic polymerase II promoter requires a functional interaction of regulatory transcriptional activators with at least one of the basal transcription factors binding in the vicinity of the TATA box . To characterize this type of interaction in vivo, we have inserted the bacterial Tet repressor-operator complex in nine different positions between an enhancer element (as-1) and the TATA box of the cauliflower mosaic virus (CaMV) 35S RNA promoter . A direct contact between the transcriptional activator ASF-1, which binds to as-1, and the transcriptional machinery should be affected by a repressor protein bound between them, as the spacing of only 34 base pairs (bp) between as-1 and the TATA box is too short to allow looping of the DNA around the repressor . In each construct, the distance of 34 bp was kept constant, while the position of the 19-bp tet operator relative to the TATA box differed by 2 bp . Thus, the position of the Tet repressor relative to the plant transcription factors was consecutively changed by 72 degrees, which allowed us to investigate whether repression depended on the stereospecific alignment of the repressor with the transcription factors . Binding of the Tet repressor to the operator blocked transcription only when the operator was inserted less tha 5 bp from the TATA box . In all other promoter derivatives, no inhibitory effect of the repressor was observed, which suggests that ASF-1 does not directly interact with the general transcription machinery. Infect Immun, 1991 Dec, 59(12), 4599 - 605 Nuclease-sensitive binding of an Actinobacillus actinomycetemcomitans leukotoxin to the bacterial cell surface; Ohta H et al.; A leukotoxin of Actinobacillus actinomycetemcomitans 301-b was solubilized from cell-associated membrane vesicles by treatment with externally added DNase and RNase and was further purified by a procedure which included ammonium sulfate fractionation, gel filtration chromatography, and ion-exchange chromatography . The purified toxin had a molecular mass of 113,000 Da by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a high isoelectric point (approximately 8.8) . From these characteristics, it was to be expected that the membrane vesicle toxin was almost identical to the leukotoxin extracted with polymyxin B in an earlier study (C.-C . Tsai, B . J . Shenker, J . M . DiRienzo, D . Malamud, and N . S . Taichman, Infect, Immun . 43:700-705, 1984) . The treatment with DNase and RNase was also highly effective for solubilizing the leukotoxin directly from whole cells, suggesting that the toxin is secreted extracellularly but retained in nucleic acids on the outermost surface of bacterial cells. Scanning Microsc, 1991 Dec, 5(4), 1129 - 33; discussion 1133-4 The effect of bacterial endotoxin upon the morphology of the tectorial membrane and stereocilia in the guinea pig cochlea; Comis SD et al.; Endotoxin of E coli was microperfused into scala tympani or injected into the cerebrospinal fluid in anaesthetised pigmented guinea pigs . The effects of endotoxin on the cochlea were studied using electrophysiological techniques and scanning electron microscopy . We found a drop in the amplitude of the cochlear microphonics and compound action potentials 2 to 2.5 hours after injection . There were also changes in the morphology of stereocilia and the tectorial membrane . The stereocilia lost their rigidity and the tectorial membrane appeared swollen . These effects were less severe in animals which were pretreated with dexamethasone. Protein Eng, 1991 Dec, 4(8), 981 - 7 High-level bacterial expression, purification and characterization of human calreticulin; Rokeach LA et al.; To investigate its cellular function and role in autoimmune disease pathogenesis, we have bacterially expressed human calreticulin, a major calcium-binding protein in the endoplasmic reticulum and a human autoantigen . This is the first report describing the heterologous expression of calreticulin from any source . The recombinant calreticulin constituted approximately 32% of the soluble Escherichia coli proteins, and was purified to apparent homogeneity by ion exchange and hydrophobic liquid chromatography . As does the bona fide protein, the recombinant calreticulin binds calcium and undergoes changes in its conformation upon Zn2+ binding . We take this as a strong indication that the folding of the E.coli-expressed calreticulin is very similar, if not identical, to that of the authentic protein . Moreover, the bacterially expressed calreticulin readily reacted with anti-human and anti-rabbit antibodies, and the anti-recombinant calreticulin antibodies immunoreacted with HeLa calreticulin . The availability of this expression system will allow us to carry out site-specific and deletion mutagenesis analysis in structure--function studies of calreticulin. Cell Struct Funct, 1991 Dec, 16(6), 495 - 502 Opposite effects of bacterial lipopolysaccharide on Fc-receptor-mediated phagocytosis of two bone marrow-derived macrophage cell lines, BDM-1 and BDM-1W3; Ohki K et al.; We have reported the isolation and characterization of three factor-dependent macrophage cell lines from bone marrow cells of C3H/HeN mice . We have since isolated a subclone, BDM-1W3, from one of these cell lines . We found previously that BDM-1W3 has a different sensitivity to bacterial lipopolysaccharide (LPS) for growth than its parental cell line, BDM-1 . In this report, we show that LPS inhibits BDM-1W3 phagocytosis of antibody-coated sheep erythrocytes (Fc-mediated phagocytosis), whereas it enhanced Fc-mediated phagocytosis by BDM-1 . It was observed that a loss of Fc-receptor capacity parallels a loss of phagocytic activity in LPS-treated BDM-1W3 cells . LPS stimulated phagocytosis of latex beads by BDM-1 and BDM-1W3, suggesting that Fc-mediated phagocytosis and phagocytosis of latex beads differ in their regulatory mechanisms . When BDM-1 cells were cultured with LPS, they underwent drastic morphological changes, whereas LPS-treated BDM-1W3 cells did not change significantly . Gamma interferon enhanced FC-mediated phagocytosis by BDM-1, while it has no significant effect on that by BDM-1W3 . These cell lines should be useful for studying signal transduction mechanisms in LPS-mediated macrophage activation. FEMS Microbiol Lett, 1991 Dec 1, 68(3), 279 - 83 Formation of demarcation zones when bacterial population waves are drawn together; Medvinsky AB et al.; Many motile bacteria (for instance, Escherichia coli) inoculated at some point in a semisolid nutrient medium can form population waves: bands or rings . The formation of these motile structures is due to chemotaxis . The population waves when they are drawn together can form two types of non-motile structures . Firstly, the population waves can collide . Secondly, in certain conditions, the waves can slow down and stop without coming into contact directly with each other . In this way demarcation zones are formed . The mechanism of the occurrence of the demarcation zones has been unknown . In this paper we show that formation of these zones is due to lack of nutrients (which at the same time act as attractants) within the narrow gap between individual bacterial populations. Differentiation, 1991 Dec, 48(3), 183 - 9 A stable cellular marker for the analysis of mouse chimeras: the bacterial chloramphenicol acetyltransferase gene driven by the human elongation factor 1 alpha promoter; Hanaoka K et al.; We have developed a method of marking of mouse cells by means of transfection of a foreign gene . The transgene chosen here was the plasmid pEF321CAT which contains the bacterial chloramphenicol acetyl transferase (CAT) gene linked to the promoter region of the human polypeptide chain elongation factor 1 alpha (hEF1 alpha) gene . Evaluation of the plasmid pEF321CAT as a cellular marker for mouse cells involved intensive examination of a transgenic mouse carrying pEF321CAT . The CAT gene was expressed in all tissues examined, demonstrating that the hEF1 alpha promoter was active in a wide range of mouse cells . The plasmid itself did not exert any harmful effect on the normal development of mice, and the CAT activity was immunohistologically detectable on sectioned tissues by the use of anti-CAT serum . When the plasmid was transferred into embryonal carcinoma (EC) cells and embryonic stem (ES) cells, the CAT gene was also found to be expressed constantly irrespective of their differentiation . These results demonstrated that the plasmid pEF321CAT can be used as a reliable and feasible cellular marker that would distinguish unequivocally the cells of each of genotype in chimeric tissues. Int J Lepr Other Mycobact Dis, 1991 Dec, 59(4), 598 - 604 Effects of cyclosporin A on bacterial growth and immunological responsiveness in BALB/c mice infected with Mycobacterium leprae; Tanaka M et al.; When BALB/c mice were infected with Mycobacterium leprae and orally treated 6 times weekly with a dose of 8 mg/kg cyclosporin A (CsA) for 19 months, the number of organisms was slightly higher at 19 months as compared with mice in which the dose of CsA was gradually decreased after 6 months and discontinued at the 8th month (p less than 0.01 for the 15th and 19th months) . Lymphocyte blast transformation (LBT) showed that spleen cells from CsA-treated mice 4 weeks after infection with M . leprae and 3 weeks after CsA treatment was stopped responded to the sonicated supernatant of M . leprae suspension (SS), M . leprae (Ml), and concanavalin A (ConA) less than those cells from mice not treated with CsA . This response was dose-dependent . At week 15, 14 weeks after CsA administration was stopped, the LBT response to SS and Ml by cells from M . leprae-infected mice exceeded that of mice without CsA treatment, and the response to ConA in M . leprae-infected mice was less than that in uninfected mice without CsA-treatment . Thus, if CsA was administered, the T-cell functions were suppressed . However, when CsA treatment was discontinued for longer periods, the T-cell function was activated . From these results, we speculate that M . leprae would have the capability of growing more abundantly in mice treated with CsA 100 mg/kg for 1 week every month. Microb Pathog, 1991 Dec, 11(6), 423 - 31 Protective activity of Bordetella adenylate cyclase-hemolysin against bacterial colonization; Guiso N et al.; Bordetella pertussis synthesizes several factors . It has been suggested that one of these factors, the adenylate cyclase-hemolysin (AC-Hly), directly penetrates target cells and impairs their normal functions by elevating intracellular cAMP . In the present study, we show that active immunization with purified B . pertussis AC-Hly or AC (a fragment of the AC-Hly molecule carrying only the adenylate cyclase activity but no toxin activity in vitro) protects mice against B . pertussis intranasal infection . Immunization with AC-Hly or AC significantly shortens the period of bacterial colonization of the mouse respiratory tract . Furthermore, B . parapertussis AC-Hly or AC are also protective antigens against B . parapertussis colonization; their protective activities are equivalent to that of the whole-cell vaccine . These results suggest that AC-Hly may play an important role in Bordetella pathogenesis, in a murine model . If this factor plays a similar role in the human disease, its use as a protective antigen could reduce not only the incidence of the disease, but also the asymptomatic human reservoir by limiting bacterial carriage. Vrach Delo, 1991 Dec, (12), 58 - 60 {A statistical analysis of the cellular and protein indices of the cerebrospinal fluid in patients with bacterial meningoencephalitis}; Iarosh OO; Clinico-liquorological comparisons were carried out in 300 patients with bacterial meningoencephalitis . The method of mathematical analysis proved able to reveal the prognostic significance of criteria of protein dispersion in the cerebrospinal fluid as one of the factors of clinical evaluation of the possible sequels of the disease. Genetics, 1991 Dec, 129(4), 1021 - 32 Ability of a bacterial chromosome segment to invert is dictated by included material rather than flanking sequence; Mahan MJ et al.; Homologous recombination between sequences present in inverse order within the same chromosome can result in inversion formation . We have previously shown that inverse order sequences at some sites (permissive) recombine to generate the expected inversion; no inversions are found when the same inverse order sequences flank other (nonpermissive) regions of the chromosome . In hopes of defining how permissive and nonpermissive intervals are determined, we have constructed a strain that carries a large chromosomal inversion . Using this inversion mutant as the parent strain, we have determined the "permissivity" of a series of chromosomal sites for secondary inversions . For the set of intervals tested, permissivity seems to be dictated by the nature of the genetic material present within the chromosomal interval being tested rather than the flanking sequences or orientation of this material in the chromosome . Almost all permissive intervals include the origin or terminus of replication . We suggest that the rules for recovery of inversions reflect mechanistic restrictions on the occurrence of inversions rather than lethal consequences of the completed rearrangement. Biotechnol Appl Biochem, 1991 Dec, 14(3), 347 - 56 Murine red blood cells as efficient carriers of three bacterial antigens for the production of specific and neutralizing antibodies; Polvani C et al.; Three bacterial toxoids, CRM 197 (mutagenized diphtheria toxin), tetanus toxoid (formaldehyde-treated tetanus toxin), and PT-9K/129G (double mutant of pertussin toxin) were encapsulated within red blood cells (RBCs) of B6D2F1 and Balb/C mice according to a mild procedure based on hypotonic dialysis-isotonic resealing that yielded undamaged RBCs . The toxoid-loaded RBCs were injected intravenously in order to immunize animals and their effects were compared to those of identical amounts (30-95 micrograms per mouse subdivided into multiple injections) of the corresponding free toxoids injected intravenously in saline . Sera from treated mice were collected and tested for titers of specific antibodies against each of the three antigens and also for titers of neutralizing antibodies, i.e., affording protection from toxic effects induced by the corresponding native toxins . In all experiments, significant seroconversion was observed with both immunization systems . Titers of both specific and neutralizing antibodies against CRM 197 and tetanus toxoid were several-fold higher upon immunization with the RBC-encapsulated toxoids, than with the free toxoids . These differences were not due to qualitatively different recognition patterns of antigenic determinants by the two types of sera . Conversely, intravenous immunization with pertussis toxoid either as RBC-encapsulated or as free antigen elicited a comparably high production of specific and of neutralizing antibodies . These data demonstrate that properly engineered RBCs behave as natural carriers and possibly adjuvants for antigens of vaccinal interest. Am J Physiol, 1991 Dec, 261(6 Pt 1), G1065 - 71 Bacterial deconjugation and enterohepatic circulation of norursocholic acid conjugates in rats; Lillienau J et al.; Experiments were performed to define the metabolism of norusocholic acid (nUC) conjugates and to quantify to what extent the bile acid pool can be enriched in these bile acids . In vitro incubations of norusocholylglycine (nUCG) and -taurine (nUCT) with small intestinal or cecal content showed deconjugation with only cecal content . Cholylglycine (CG) was deconjugated by small intestinal and cecal content . Infusion of nUCG and CG showed that only a small proportion of nUCG was deconjugated after 24 h of enterohepatic circulation, whereas all CG was deconjugated . When nUCT was administered orally, deconjugation was shown to take place mainly in the cecum . Chronic feeding of nUCT enriched the bile acid pool with only 20% nUCT . We conclude that nUC conjugates are deconjugated primarily by bacteria in the cecum and colon, in contrast to CG, which, in addition to cecum and colon, is deconjugated in the distal small intestine . nUCT and its metabolites do not enrich in the circulating bile acid pool mainly for the following reasons: 1) nUC conjugates have a low affinity for the ileal transport system; 2) nUC, even if formed by deconjugation, is not passively absorbed at a sufficient rate; 3) the small amount of norursodeoxycholic acid formed from nUC is glucuronidated in the liver and glucuronide conjugates do not undergo enterohepatic circulation; and 4) nUC conjugates do not suppress bile acid biosynthesis. Toxicol Lett, 1991 Dec, 59(1-3), 187 - 95 Binding of the strong bacterial mutagen, 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX) to bovine serum albumin; Haataja L et al.; Binding of 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX) to bovine serum albumin (BSA) was studied . MX bound mainly reversibly to BSA but, for a minor part, also irreversibly . It was possible to extract the main part of the reversibly bound MX with ethyl acetate and the extractable compound was chromatographically identical to MX . The affinity-binding characteristics of the interaction with albumin were K = 1.6 x 10(7) M-1, n = 3.4 . Furthermore, mutagenicity studies indicated that reversibly bound MX remained mutagenic but that irreversibly bound MX was no longer mutagenic in the Ames test . These results suggest that the binding of MX to albumin is an important factor for both the toxicological effects and the toxicokinetics of MX. Am J Physiol, 1991 Dec, 261(6 Pt 2), R1431 - 7 Mechanisms involved in the rapid dissipation of plasma epinephrine response to bacterial endotoxin in conscious rats; Qi M et al.; Conscious rats treated with bolus endotoxin (ET; 4.0 mg/kg) increased plasma epinephrine (Epi) 48-fold (from 134 +/- 5 to 6,545 +/- 1,607 pg/ml) at 30 min, but by 6 h this elevation was less than 9-fold above control (1,174 +/- 166 pg/ml) . In contrast, plasma norepinephrine (NE) elevation (6.5-fold above control) was maintained during the protocol . The present study was designed to test the hypothesis that the decreased Epi response following ET was due to 1) depletion of adrenal Epi content such that adrenomedullary stimulation would not release Epi, 2) decreased Epi release with direct stimulation, i.e., desensitization of release, or 3) decreased afferent signals generated by ET itself . In these experiments an initial low ET dose (0.5 mg/kg) was followed 3 h later by a second dose of either the same (0.5 mg/kg) or greater (4.0 mg/kg) magnitude . Plasma Epi was elevated following the initial (low) dose but not following the same second (0.5 mg/kg) dose, whereas the second higher dose (4.0 mg/kg) resulted in elevated plasma Epi . This response to high dose was 60% less than that observed with 4.0 mg/kg as an initial dose . Just before the second ET bolus, there was no depletion of adrenal Epi content (P greater than 0.05 saline vs . ET treated), and direct nerve stimulation demonstrated enhanced rather than attenuated Epi release from the adrenal medulla (P less than 0.05 saline vs . ET treated) . These results suggest that the decline in Epi following the ET-induced elevation may be mediated by a decrease in the afferent signal that initiates Epi release. Carcinogenesis, 1991 Dec, 12(12), 2305 - 9 Comparison of the inactivation of mammalian and bacterial O6-alkylguanine-DNA alkyltransferases by O6-benzylguanine and O6-methylguanine; Dolan ME et al.; The inactivation of human and Escherichia coli O6-alkylguanine-DNA alkyltransferase by O6-methylguanine and O6-benzylguanine was compared . When HT29 cell extracts or E . coli Ada protein were incubated in the presence of 200 microM O6-methylguanine for 1 h, alkyltransferase activity was reduced to 44 and 39% of control levels respectively . However, under the same conditions O6-benzylguanine completely depleted alkyltransferase activity in the extract from human cells but had virtually no effect on the Ada protein . Incubation of the HT29 cell alkyltransferase with O6-benzyl{3H}guanine resulted in a time-dependent production of {3H}guanine . No similar production of {3H}guanine was observed in the presence of the Ada protein . In CHO cells transfected with the bacterial ada gene (CHO-ada) or the human alkyltransferase cDNA (CHO-MGMT), treatment with 500 microM O6-methylguanine inhibited both alkyltransferases by greater than 85% . In contrast, 2 microM O6-benzylguanine inhibited human alkyltransferase expressed in CHO-MGMT cells by greater than 99% though concentrations as high as 25 microM for 24 h had no inhibitory effects on the bacterial alkyltransferase expressed in CHO-ada cells . This selective inhibition was also observed in vivo in transgenic mice expressing ada in the liver where O6-benzylguanine caused a decrease of only 40% in total hepatic alkyltransferase activity compared to 95% in non-transgenic mice, consistent with inhibition of only the mammalian alkyltransferase and maintenance of bacterial alkyltransferase activity in these animals . Thus, while O6-methylguanine at high concentrations inactivates both bacterial and mammalian alkyltransferases, O6-benzylguanine is a substrate only for the mammalian protein and is unable, perhaps due to steric hindrance, to inhibit the Ada protein. Ann Surg, 1991 Dec, 214(6), 719 - 23 The effect of glucocorticoid administration on bacterial translocation . Evidence for an acquired mucosal immunodeficient state; Alverdy J et al.; Adherence of bacteria to intestinal epithelial cells may be the crucial initiating event for translocation and is normally prevented by both specific (secretory IgA) and nonspecific (mucus, bacterial antagonism, desquamation) mucosal defense mechanisms . The purpose of this study was to examine the effect of dexamethasone administration on mucosal immunity; specifically bacterial adherence and IgA . Twenty Fischer rats were randomly assigned to two groups of 10 animals each . Group I received 0.5 mL saline injection intraperitoneally (IP); and group II, 0.8 mg/150 g body weight dexamethasone IP per day for 2 consecutive days . The cecum mesenteric lymph nodes, and bile were aseptically collected, and bacterial adherence, bacterial translocation, and IgA concentration were determined . Results indicate that, compared with saline-treated animals, dexamethasone-treated animals had a fall in IgA (54 +/- 24 versus 232 +/- 41 micrograms/mg protein), an increase in bacterial adherence (8.2 +/- 0.5 versus 3.4 +/- 0.6 cfu (log10)/g cecum), and an increased incidence of bacterial translocation to the mesenteric lymph nodes (60% versus 0%) . These data suggest that glucocorticoids may promote bacterial translocation by impairment of mucosal IgA synthesis. Oral Microbiol Immunol, 1991 Dec, 6(6), 332 - 40 Distribution and immunochemical specificities of fimbriae of Porphyromonas gingivalis and related bacterial species; Ogawa T et al.; Rabbit polyclonal antibody (Poly-1) and mouse monoclonal antibodies (mAbs) TO-11, TO-14 and TO-M1 specific for Porphyromonas gingivalis 381 fimbriae were prepared . Poly-1 and the 3 mAbs were screened for their reactivity with whole cells oral and nonoral black-pigmented bacterial species by enzyme-linked immunosorbent assay (ELISA) and the binding experiment using {125I}Poly-1 and {125I}mAbs . ELISA revealed that Poly-1 definitely reacted with whole cells of all the 11 strains of P . gingivalis tested . However, 8 of 11 P . gingivalis strains reacted with mAbs TO-11, TO-14 and TO-M1 . These results were confirmed by the specific binding of radiolabelled Poly-1 and mAb TO-11 to the 8 strains . The M(r) of the fimbrial subunit protein (fimbrilin) isolated and purified from P . gingivalis strains 381, BH18/10, HW24D-1, 6/26 and OMZ 314 was 41 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis . It was found by immunoblotting that mAbs TO-11 and TO-14/TO-M1 recognized different epitopes of fimbrial protein from P . gingivalis strains . Immunoelectron micrographs of whole cells and the purified fimbriae of P . gingivalis strains visualized similar serotype-specific antibody bindings to the fimbriae . These results indicate that 11 strains of P . gingivalis could be divided into at least 2 separate groups based on the immunochemical specificities of the fimbriae. Infect Dis Clin North Am, 1991 Dec, 5(4), 753 - 79 Bacterial endotoxin: molecular relationships between structure and activity; Rietschel ET et al.; The significance of endotoxins in bacterial infection and their role as bacterial surface antigens (O antigens) have stimulated investigations into their chemical nature and the mechanisms of their biologic action during the last few decades . This article summarizes some of the recent results and emphasizes structure-activity relationships. Mutat Res, 1991 Dec, 261(4), 237 - 48 Mutagenicity spectra in bacterial strains of airborne and engine exhaust particulate extracts; Crebelli R et al.; The mutagenicity spectra of the organic extracts of both airborne particulate matter and diesel and gasoline soot particles were determined using a battery of 9 bacterial strains of different genetic specificity . The assays with crude extracts and with fractionated acidic, neutral and basic components revealed striking differences in the patterns of mutagenic responses produced by each of the complex mixtures investigated . The mutagenicity of air particulate matter was shown to depend mainly on direct-acting acidic and neutral compounds, with a lesser contribution of basic promutagens which required exogenous metabolic activation by liver S9 . The assays with a diesel soot extract indicated the prevailing contribution of direct-acting acidic and neutral compounds, and suggested an important role also for nitro derivatives other than nitropyrenes . The gasoline exhaust was characterized by powerful promutagenic compounds, belonging to either the acidic, neutral or basic fractions . The implications of these results are discussed with respect to the contribution of engine exhausts to air pollution, and the possible use of mutagenicity spectra in the analysis of environmental complex mixtures. Gastroenterology, 1991 Dec, 101(6), 1656 - 62 Spontaneous bacterial peritonitis in patients with hepatitis B-related cirrhosis and hepatocellular carcinoma; Wang SS et al.; To delineate the natural clinical course of spontaneous bacterial peritonitis in hepatitis B-related cirrhosis and to determine if it occurs in hepatocellular carcinoma, a prospective survey was conducted in 262 patients over 2 1/2 years . The in-hospital incidence and mortality rates of spontaneous bacterial peritonitis were 21.6% and 36.4%, respectively, in cirrhosis and 7.3% and 50% in hepatocellular carcinoma . In cirrhosis, the cumulative probability of annual recurrence of spontaneous bacterial peritonitis was 47.3%, which was significantly higher than the annual probability of occurrence of 11.3% in those with no previous attack (P less than 0.0001) . The cumulative probability of annual survival was 27.6% in the spontaneous bacterial peritonitis patients, significantly lower than the probability of 64.0% in the control group (P = 0.0001) . A univariate analysis, with Kaplan-Meier curves compared by the Mantel-Cox test, and subsequent multivariate analysis by stepwise Cox regression procedure were used to evaluate 37 variables recorded immediately after admission . Blood urea nitrogen concentration greater than 10.5 mmol/L urea (greater than 30 mg/dL) and ascitic fluid protein concentration less than 7.35 g/L (less than 735 mg/dL) were found to be the only significant predictors of lower annual survival; ascitic fluid protein concentration less than 7.50 g/L (less than 750 mg/dL) was the only significant predictor of higher annual recurrence . The authors conclude that spontaneous bacterial peritonitis has a high risk of recurrence in hepatitis B-related cirrhosis and that the same disease occurring in patients with hepatocellular carcinoma is related to the underlying cirrhosis rather than the hepatocellular carcinoma. Nucleic Acids Res, 1991 Nov 25, 19(22), 6119 - 22 Levels of tRNAs in bacterial cells as affected by amino acid usage in proteins; Yamao F et al.; Transfer RNAs of Mycoplasma capricolum were separated by two-dimensional polyacrylamide gel electrophoresis, and the relative abundance of each of the 28 known tRNA species was measured . There existed a correlation between the relative amount of isoacceptor tRNAs and the frequency in choosing synonymous codons that could be translated by the isoacceptors . Furthermore, it was observed that the total amount of tRNAs for a particular amino acid was paralleled by the composition of the amino acid in ribosomal proteins . A similar relationship was obtained from reexamination of the previous data on Escherichia coli tRNAs, suggesting that the amount of tRNAs for an amino acid is affected by the usage of the amino acid in proteins. J Biol Chem, 1991 Nov 25, 266(33), 22671 - 7 The yeast mitochondrial RNA polymerase specificity factor, MTF1, is similar to bacterial sigma factors; Jang SH et al.; We have purified the protein that confers selective promoter recognition on the core subunit of the yeast mitochondrial RNA polymerase . The N-terminal sequence of the 43-kDa specificity factor identified it as the product of the MTF1 gene described by Lisowsky and Michaelis (1988) . We confirmed that MTF1 encoded the specificity factor by analyzing extracts from a yeast strain bearing a disruption of the gene . The extracts contained normal levels of core RNA polymerase but lacked selective transcription activity; adding the purified 43-kDa protein restored selective transcription . Comparison of the MTF1 protein sequence to the family of bacterial sigma factors has revealed striking similarity to domains identified with--10 promoter recognition, promoter melting, and holoenzyme stability. J Biol Chem, 1991 Nov 25, 266(33), 22319 - 22 Expression of duck lens delta-crystallin cDNAs in yeast and bacterial hosts . Delta 2-crystallin is an active argininosuccinate lyase; Barbosa P et al.; The major soluble protein in the lenses of most birds and reptiles is delta-crystallin . In chickens and ducks the delta-crystallin gene has duplicated, and in the duck both genes contribute to the protein in the lens, while in the chicken lens there is a great preponderance of the delta 1 gene product . Purified delta-crystallin has previously been shown to possess the enzymatic activity of argininosuccinate lyase . In order to determine the enzymatic properties of the two duck delta-crystallins their corresponding cDNA molecules were placed in yeast and bacterial expression plasmids . In Saccharomyces cerevisiae, the activity of each crystallin was assessed by transformation of the expression plasmids into a strain deficient for argininosuccinate lyase activity . The ability of the resulting yeast to grow on arginine deficient medium was used as a measure of enzymatic activity . Yeast expressing the duck delta 2-crystallin protein grew rapidly, while those expressing delta 1-crystallin failed to grow . Enzyme activity measurements confirmed the presence of activity in the delta 2-crystallin-expressing yeast, and no detectable activity could be demonstrated in the delta 1-crystallin-expressing yeast . Northern blotting of RNA from the transformed yeast revealed equal levels of mRNA species from the two constructs . For further analysis, the delta 2-crystallin cDNA was placed in the bacterial expression plasmid, pET-3d . The delta 2-crystallin protein produced in Escherichia coli was purified to homogeneity and analyzed to determine the kinetic properties . A Km of 0.35 mM was determined for argininosuccinate and a Vm of 3.5 mumols/min/mg was determined . These data demonstrate that, following duplication of the primordial argininosuccinate lyase gene, one of the genes maintained its role as an enzyme (delta 2-crystallin) while also serving as a crystallin and the other has evolved to specialize as a structural protein in the lens (delta 1-crystallin), presumably losing most or all of its catalytic capacity. Brain Res, 1991 Nov 22, 565(1), 85 - 93 Production of the neuronal growth-associated protein GAP-43 in a bacterial expression system; Schuh SM et al.; GAP-43, a major protein of neuronal growth cones and certain presynaptic terminals, is a candidate for important functions in both axon growth and synaptic plasticity . To facilitate studies that may elucidate these functions, we have efficiently generated large quantities of GAP-43 by introducing a GAP-43 cDNA into a bacterial expression system driven by T7-RNA polymerase . Two constructs were expressed in this system: one (pT7Ava-GAP) produces a fusion protein in which the first 16 amino acids of GAP-43 are replaced by 11 amino acids of the phage T7 capsid protein; the other (pT7FL-GAP) produces full length GAP-43 . After the bacteria were lysed, both products were soluble, and could be efficiently purified by HPLC chromatography on a C4 reversed-phase column . One liter of bacterial culture yielded 50 mg of purified fusion protein or 10 mg of complete GAP-43 . When it was incubated with protein kinase C, the fusion protein was phosphorylated at the same single site (serine 41) that is phosphorylated in cultured neurons . The ability to produce large quantities of GAP-43 by this procedure should expedite future studies investigating its structure, posttranslational modification, and function. N Engl J Med, 1991 Nov 21, 325(21), 1461 - 7 Effect of octreotide on intestinal motility and bacterial overgrowth in scleroderma; Soudah HC et al.; BACKGROUND . Patients with scleroderma may have abnormal motility of the small intestine, with pseudoobstruction and bacterial overgrowth . Standard stimulatory agents are often ineffective in such patients . Because the somatostatin analogue octreotide evokes intestinal motor activity in normal subjects, we hypothesized that it might increase motility in patients with scleroderma . METHODS . We studied the effects of octreotide on intestinal motility and plasma motilin concentrations in five fasting patients with scleroderma who had bacterial overgrowth and in six fasting normal subjects . The motor effects of octreotide were correlated with its effects on abdominal symptoms and bacterial overgrowth as determined by the level of breath hydrogen excretion . RESULTS . In the normal subjects, octreotide (10 micrograms subcutaneously) increased the mean (+/- SD) frequency of intestinal migrating complexes, which reflect intestinal motility, from 1.5 +/- 1.0 to 4.1 +/- 1.1 every three hours . In the patients with scleroderma, who had no spontaneous migrating complexes, octreotide (100 micrograms) induced 3.6 +/- 2.3 complexes every three hours . These complexes propagated at the same velocity and had two-thirds the amplitude of the spontaneous complexes in normal subjects . Plasma motilin concentrations, which were higher in the patients with scleroderma (229 +/- 74 pmol per liter) than in the normal subjects (112 +/- 37 pmol per liter), were inhibited by octreotide, suggesting that intestinal activity evoked by octreotide is independent of motilin . Treatment of the patients with scleroderma with octreotide (50 micrograms every evening) for three weeks reduced breath hydrogen excretion while they were fasting from 25 +/- 5 to 4 +/- 2 ppm (P = 0.001) and breath hydrogen excretion after they ingested 50 g of glucose from 46 +/- 24 to 8 +/- 7 ppm (P = 0.015); these reductions were accompanied by a significant decrease in nausea, bloating, and abdominal pain and by less frequent emesis . CONCLUSIONS . Octreotide stimulates intestinal motility in normal subjects and in patients with scleroderma . In such patients, the short-term administration of octreotide reduces bacterial overgrowth and improves abdominal symptoms . This agent may be useful for the treatment of intestinal dysmotility in patients with scleroderma. Biochim Biophys Acta, 1991 Nov 15, 1080(3), 215 - 20 Bacterial catalase-peroxidases are gene duplicated members of the plant peroxidase superfamily; Welinder KG; Bacterial catalase-peroxidases are enzymes containing 0.5-1.0 heme per subunit . The identical subunits are generally 80 kDa in size, and the sequenced subunits of E . coli, S . typhimurium and B . stearothermophilus contain 726-731 amino acid residues per subunit . The heme-containing peroxidases of plants, fungi and yeast are monomeric, homologous and 290-350 residues in size . Analyses of the amino acid sequences indicate that the double length of the bacterial peroxidases can be ascribed to gene duplication . Each half is homologous to eukaryotic, monomeric peroxidase and can be modelled into the high-resolution crystal structure of yeast cytochrome c peroxidase . The comparisons and modelling have predicted: (1) the C-terminal half does not bind heme, and bacterial peroxidases have one heme per subunit; (2) the ten dominating helices observed in the yeast enzyme are highly conserved and connected by surface loops which are often longer in the bacterial peroxidases; and (3) yeast cytochrome c peroxidase has evolved more slowly than other known peroxidases . The study has revealed ten invariant residues and a number of highly conserved residues present in peroxidases of the plant peroxidase superfamily and provides a basis for rationally engineered peroxidases. Eur J Biochem, 1991 Nov 15, 202(1), 161 - 5 Expression of rat heme oxygenase in Escherichia coli as a catalytically active, full-length form that binds to bacterial membranes; Ishikawa K et al.; A plasmid, pKK-RHO, was constructed by incorporating the coding sequence of a cDNA for rat heme oxygenase into the expression vector pKK233-2 . Escherichia coli strain XL1-blue transformed with pKK-RHO produced a catalytically active, full-length heme oxygenase . The 32-kDa native enzyme expressed, was localized in the bacterial membranes, possibly due to the spontaneous membrane-binding properties of a hydrophobic segment in its C-terminal region . During cultivation, a few degraded forms of heme oxygenase that had lost their membrane-associative properties appeared . Probably, some bacterial proteases cut the native heme oxygenase at sites near its C-terminus and so release hydrophilic peptides of heme oxygenase from the membranes . A 30-kDa polypeptide, one of the degraded forms of heme oxygenase, retained ability to accept electrons from NADPH--cytochrome P450 reductase and also activity for catalyzing breakdown of heme to biliverdin . The cultured cells were pale green . From them we extracted green pigment(s), of which the absorption spectrum closely resembled that of biliverdin, suggesting that a large amount of the endogenous heme of E . coli was actually degraded to biliverdin by the expressed heme oxygenase. Anal Biochem, 1991 Nov 15, 199(1), 68 - 74 The compositional analysis of bacterial extracellular polysaccharides by high-performance anion-exchange chromatography; Clarke AJ et al.; A high-performance liquid chromatography (HPLC) method with pulsed-amperometric detection (PAD) was developed for the compositional analysis of the acidic, neutral, and basic monosaccharides recovered from the acid hydrolysis of bacterial cell wall polysaccharides . This HPLC-PAD method involved the chromatography of the acid hydrolysis products on a CarboPac PA-1 anion-exchange column of pellicular resin, with PAD detection following postcolumn addition of alkali . Complete resolution of a mixture of 19 monosaccharides, comprising 9 neutral, 3 basic, and 7 acidic sugars, frequently found in bacterial polysaccharides was achieved within 60 min by the system . The presence of amino acids in the mixture was shown not to affect the analysis . This protocol was applied to the compositional analysis of 2 extracellular polysaccharides produced by Escherichia coli, colanic acid, and K30 antigen, which share constituent monosaccharides . The overproduction of extracellular polysaccharide in E . coli CWG56 was shown to be a consequence of deregulation of K30 biosynthesis and not of coexpression of an additional polymer. FEBS Lett, 1991 Nov 4, 292(1-2), 9 - 12 The aromatic amino acid content of the bacterial chaperone protein groEL (cpn60) . Evidence for the presence of a single tryptophan; Price NC et al.; Studies of the absorption and fluorescence properties of the chaperone protein groEL (cpn60) from Escherichia coli show that tryptophan is present, in contrast to the proposed amino acid sequence of the protein (Hemmingsen, S.M . et al . (1988) Nature 333, 330-334) . By determining a suitable value for the specific absorption coefficient of the protein at 280 nm, it has been shown that the content of the aromatic amino acids corresponds to a single tryptophan and (most probably) seven tyrosines per subunit (Mr 57,200). APMIS, 1991 Nov, 99(11), 1049 - 54 Mobiluncus species in bacterial vaginosis: aspects of pathogenesis; Moi H et al.; Mobiluncus is an anaerobic motile rod associated with bacterial vaginosis . In this work, luminol-enhanced chemiluminescence was used to study the ability of Mobiluncus spp . from the vaginas of women with bacterial vaginosis to induce, in the presence of normal adult serum, oxidative metabolism of polymorphonuclear leukocytes, which is an indirect measure of phagocytic activity . M . curtisii induced a significantly (p less than 0.05) lower response than M . mulieris, which indicates that M . curtisii escapes phagocytosis more easily . Indirect immunofluorescence assays showed IgG antibodies to M . curtisii at significantly (p less than 0.01) higher titres than to M . mulieris in women with bacterial vaginosis . The titres were higher in patients with bacterial vaginosis than in women without vaginosis and healthy men . No antibodies to Mobiluncus spp . of secretory IgA type were found in vaginal washings . These results indicate that M . curtisii is a more virulent species than M . mulieris, and agree with reports of M . curtisii found in postoperative and extragenital infections. South Med J, 1991 Nov, 84(11), 1412 - 3 Spontaneous bacterial peritonitis during bowel preparation: an example of clinical translocation; Poole GV; During oral colonic lavage in preparation for colonoscopy, a patient had peritonitis in the absence of visceral perforation . Spontaneous peritonitis after mechanical bowel preparation has not been reported previously, and must be extremely rare . I speculate that it was due to translocation of bacteria from the gut to the peritoneal cavity. J Nutr, 1991 Nov, 121(11), 1860 - 8 Influence of intake of skim milk from cows immunized with intestinal bacterial antigens on onset of renal disease in (NZB x NZW)F1 mice fed ad libitum or restricted in energy intake; Murosaki S et al.; Autoimmune disease-prone (NZB x NZW)F1 (B/W) female mice are a model of human lupus erythematosus . Immune milk, obtained from cows immunized with various intestinal bacterial antigens, was given to B/W mice as a component of diets beginning at 8 wk of age . Diets were fed ad libitum or restricted to 60% of ad libitum energy intake . Controls were fed commercial skim milk . In B/W mice fed ad libitum, the titers of anti-single-stranded DNA antibodies were significantly lower in immune milk-fed mice at 4 and 6 mo of age . Onset of proteinuria was delayed and life span was significantly prolonged by immune milk feeding . Surface phenotypes of the T cells and levels of the responsiveness of lymphocytes to mitogens were not changed by immune milk feeding . The B/W mice restricted to 60% of ad libitum energy intake, which preserved immune responsiveness, had not developed proteinuria by 14 mo of age, irrespective of immune milk feeding or control milk feeding . However, at 10 mo of age, the level of plasma antibodies against intestinal bacteria was significantly higher in energy-restricted mice fed control milk than in those fed immune milk or in mice fed ad libitum. Circ Res, 1991 Nov, 69(5), 1196 - 206 Polymorphonuclear neutrophil contribution to induced tolerance to bacterial lipopolysaccharide; Barroso-Aranda J et al.; The objective of this study was to investigate mechanisms by which polymorphonuclear neutrophils (PMNs) contribute to the tolerance induced by repeated lipopolysaccharide (LPS) injections . Tolerance was developed by daily intraperitoneal injections of sublethal doses of LPS for 4 days (LPS-tolerant group); controls were not pretreated (LPS-control group) . Both groups were challenged with 9 mg/kg i.v . Escherichia coli LPS, a dose that resulted in 25% survival in LPS-control rats compared with 100% survival in LPS-tolerant rats . LPS injection caused an initial neutropenia in both groups . The neutropenia persisted throughout the experiment in LPS-control rats, whereas in LPS-tolerant rats the circulating PMN count increased dramatically; after 6 hours, the PMN count was 16-fold higher than that in LPS-control rats . Activation of circulating PMNs, PMN adhesion to nylon fibers, and tumor necrosis factor/cachectin activity were all increased in control rats given LPS . In contrast, LPS-tolerant rats had low activation of circulating PMNs, no trend for PMN adhesion to nylon fibers, and markedly reduced tumor necrosis factor activity . To determine whether neutropenia was associated with a trapping of PMNs in the microcirculation, we used a carbon perfusion technique 6 hours after LPS injection and examined histological sections of the myocardium . All of the arterioles and venules in both groups contained carbon; only capillaries showed evidence of obstruction . A significantly higher percentage of obstructed capillaries was observed in LPS-control rats than in LPS-tolerant rats . Obstruction of capillaries was consistently associated with trapped leukocytes . We conclude that PMN cytotoxicity induced by LPS involves microcirculatory entrapment and activation of PMNs . Repeated LPS pretreatment reduces dramatically circulating PMN activation and adhesion and is associated with an elevated circulating PMN count, a low degree of microvascular plugging, and survival after a normally lethal dose of LPS. Plant Cell, 1991 Nov, 3(11), 1187 - 93 Control of ethylene synthesis by expression of a bacterial enzyme in transgenic tomato plants; Klee HJ et al.; Synthesis of the phytohormone ethylene is believed to be essential for many plant developmental processes . The control of ripening in climacteric fruits and vegetables is among the best characterized of these processes . One approach to reduce ethylene synthesis in plants is metabolism of its immediate precursor, 1-aminocyclopropane-1-carboxylic acid (ACC) . Soil bacteria containing an enzyme, ACC deaminase, were identified by their ability to grow on ACC as a sole nitrogen source . The gene encoding ACC deaminase was cloned and introduced into tomato plants . Reduction in ethylene synthesis in transgenic plants did not cause any apparent vegetative phenotypic abnormalities . However, fruits from these plants exhibited significant delays in ripening, and the mature fruits remained firm for at least 6 weeks longer than the nontransgenic control fruit . These results indicated that ACC deaminase is useful for examining the role of ethylene in many developmental and stress-related processes in plants as well as for extending the shelf life of fruits and vegetables whose ripening is mediated by ethylene. Biofizika, 1991 Nov-Dec, 36(6), 1043 - 7 {Structure of an Escherichia coli bacterial suspension}; Kakorin SA et al.; The structure of bacterial suspensions of Escherichia coli M-17 at the counting concentrations of the cells 10(7), 10(8), 10(9) i/ml and in the temperature range of (18-50) degrees C has been investigated by means of orientational conductometric, electron microscopic and UV-spectroscopic methods . On the basis of experimental relationships of the anisotropy of suspensions electric conductivity upon the intensity of a sinusoidal electric field and relaxation of anisotropy after switching off the field the function of the distribution of bacteria with respect to their sizes was evaluated at different temperatures and concentrations . The conductometric function of bacteria distribution is in a good agreement with the analogous function obtained with the help of the electron microscope . In accordance with the functions the suspension of E . coli contained three kinds of cells: high electronic density, low electronic density bacteria and bacteria aggregates . Relative amounts of every kind of bacteria depended on temperature and concentration of cells . The minimum of bacteria aggregates and maximum of low electronic density cells were obtained in the temperature range of (32-42) degrees C . This fact could be explained by the activation of the transport membrane systems in this temperature range . This hypothesis was confirmed by the UV-spectroscopic method. Vrach Delo, 1991 Nov, (11), 69 - 73 {The use of a method of mathematical analysis in studying the permeability of the hemato-encephalic barrier in bacterial meningoencephalitis}; Iarosh OA; The authors studied the blood-brain barrier permeability in 70 patients depending on the severity and terms of bacterial meningoencephalitis . It was shown that employment of the probability-statistical analysis of quantitative content of metal in the blood and cerebrospinal fluid allows to evaluate the working regimen of the blood-brain barrier from initial manifestations of functional insufficiency to failure of barrier compensation systems. Ophthalmology, 1991 Nov, 98(11), 1703 - 10 Is chronic intraocular inflammation after lens implantation of bacterial origin? Cusumano A, Busin M, Spitznas M. In an effort to better understand the cause of chronic intraocular inflammation after intraocular lens (IOL) implantation, both scanning and transmission electron microscopy were used to compare 1 anterior chamber, 1 iris-fixated, and 3 posterior chamber IOLs removed for this condition between 2 and 16 months after implantation with 8 IOLs explanted for other reasons (decentration in 4 cases, bullous keratopathy in 4 cases) . Colonization with non-slime-producing, as well as slime-producing bacteria (1 case) in the presence of a thin membranous structure of cellular origin (multinucleated giant cells and macrophage-like cells) was demonstrated on all of the 5 IOLs explanted from inflamed eyes . Neither bacteria nor membranous structures could be identified on the IOLs removed because of dislocation or from eyes with bullous keratopathy . These observations indicate that bacterial colonization and the consequent host response may be characteristic features of many otherwise unexplained cases of intraocular inflammation after IOL implantation. Eur J Clin Microbiol Infect Dis, 1991 Nov, 10(11), 966 - 9 Determination of D-lactate concentration for rapid diagnosis of bacterial infections of body fluids; Marcos MA et al.; The value of determining D-lactate concentrations in body fluids for the rapid diagnosis of bacterial infections was investigated . A total of 336 body fluid samples were analyzed: 208 ascitic fluids, 57 synovial fluids, 40 cerebrospinal fluids and 32 pleural fluids . Using a cut-off value of 0.05 mM, the overall sensitivity was 0.96 and the specificity was 0.88 . Therefore, the measurement of D-lactate concentration in body fluids offers a rapid (2-hour) and useful method of differentiating between infectious and non-infectious body fluid diseases. Appl Environ Microbiol, 1991 Nov, 57(11), 3171 - 7 Biochemical fingerprinting of water coliform bacteria, a new method for measuring phenotypic diversity and for comparing different bacterial populations; Kuhn I et al.; A simple, automated microplate system for biochemical characterization of water isolates can be used to obtain fingerprints of the bacterial flora from various water samples . Mathematical models for calculating the diversities and similarities between bacterial populations are described for such fingerprints . The diversity may give information on whether an indigenous or allochthonous flora is present, and the similarities between bacterial populations, as calculated by using a population similarity coefficient (Sp), may indicate contaminations between different water samples . The system was demonstrated on coliform bacterial populations from various water samples, with or without suspected intercontamination . For unrelated water samples, the Sps were close to 0, whereas repeated samples of the same source showed Sps of 0.64 to 0.74 . The Sp values from several water samples were also clustered to form a dendrogram, thus indicating the relative similarities between the bacterial populations to confirm suspected common sources of pollution. Appl Environ Microbiol, 1991 Nov, 57(11), 3144 - 9 Mineralization of the sulfonated azo dye Mordant Yellow 3 by a 6-aminonaphthalene-2-sulfonate-degrading bacterial consortium; Haug W et al.; Under anaerobic conditions the sulfonated azo dye Mordant Yellow 3 was reduced by the biomass of a bacterial consortium grown aerobically with 6-aminonaphthalene-2-sulfonic acid . Stoichiometric amounts of the aromatic amines 6-aminonaphthalene-2-sulfonate and 5-aminosalicylate were generated and excreted into the medium . After re-aeration of the culture, these amines were mineralized by different members of the bacterial culture . Thus, total degradation of a sulfonated azo dye was achieved by using an alternating anaerobic-aerobic treatment . The ability of the mixed bacterial culture to reduce the azo dye was correlated with the presence of strain BN6, which possessed the ability to oxidize various naphthalenesulfonic acids . It is suggested that strain BN6 has a transport system for naphthalenesulfonic acids which also catalyzes uptake of sulfonated azo dyes . These dyes are then gratuitously reduced in the cytoplasm by unspecific reductases. Monatsschr Kinderheilkd, 1991 Nov, 139(11), 783 - 5 {Nasal encephalocele as a cause of recurrent bacterial meningitis}; Kohler T et al.; Nasal encephaloceles are rare inborn malformations of the central nervous system . We report on a 30 months old boy with recurrent bacterial meningitis where a nasal encephalocele was diagnosed . We describe diagnostic procedures and the development and treatment of this interesting disease . The nasal encephalocele was successfully removed surgically. Exp Physiol, 1991 Nov, 76(6), 979 - 82 Segmental heterogeneity of rat colonic electrogenic secretion in response to the bacterial enterotoxin Escherichia coli STa in vitro; Young A et al.; The effects of the bacterial toxin Escherichia coli STa on electrogenic secretion in proximal, mid and distal rat colon were examined in vitro using the short-circuit current (Isc) as an index of secretion . All areas showed toxin-elevated Isc which was reduced by bumetanide . The proximal colon was the most sensitive (lowest ED50) but displayed the lowest maximal increase in Isc, the mid colon had the largest maximal electrogenic response while the distal colon proved to be the least sensitive (highest ED50) . Atropine (1 microM) and tetrodotoxin (1.25 microM) reduced the maximum Isc response to STa by up to 42%, indicating that the enteric nervous system may be partly involved in the response. J Appl Bacteriol, 1991 Nov, 71(5), 460 - 6 Analysis of quantal response data from bacterial destruction studies in sterilization: the Stumbo estimate is biased; Hachigian I et al.; Real and simulated experimental data and theoretical data from quantal response experiments were used to make a comparison between the analysis of data from a quantal response experiment and data from a direct enumeration experiment . The method of analysis for each is differentiated, thereby enhancing the utility of the quantal response experiment in sterilization studies . From this comparison it appears that the Stumbo estimate of the D value is biased . Furthermore, the Stumbo estimate depends upon the spore load per replicate in quantal response type experiments, which makes experimental comparisons difficult . Another estimate of D is demonstrated which overcomes some of these shortcomings. Int J Pancreatol, 1991 Nov-Dec, 10(3-4), 229 - 36 The resorption of FITC-dextran 10,000 from the peritoneum in different modifications of bile-induced acute pancreatitis and in bacterial peritonitis; Tarpila E et al.; The resorption from the peritoneum of fluorescein-isothiocyanate-conjugated (FITC) dextran with a mol wt of 10,000 was studied after 6, 15, and 24 h in rats with (1) only laparotomy (LC), (2) bacterial peritonitis (BP), (3) bile-induced acute pancreatitis (AP), (4) acute pancreatitis induced with contaminated bile (AIP), and (5) cerulein administration during acute pancreatitis (CAP) . Animals in the AIP and CAP groups had a significantly higher mortality rate at 24 h and higher hematocrit at 6 h, indicating severe disease in these animals . At 6 and 15 h, all groups displayed similar peritoneal resorption . After 24 h, all groups with active inflammation showed significantly higher resorption than laparotomy controls . We conclude that peritoneal resorption as defined is independent of the severity and mode of induction of acute experimental pancreatitis and that it is the same as in bacterial peritonitis. Br J Urol, 1991 Nov, 68(5), 473 - 8 Characteristics of mast cells in normal bladder, bacterial cystitis and interstitial cystitis; Christmas TJ et al.; An analysis was made of the numbers and characteristics of mast cells in lateral bladder wall biopsies from 22 patients with interstitial cystitis, 6 with bacterial cystitis and 8 normal controls, using toluidine blue stains and computerised video image analysis techniques . A significantly greater number of mast cells were found within the detrusor muscle in interstitial cystitis than in bacterial cystitis or normal controls . Within the urothelium and submucosa, mast cell numbers were significantly greater than in normal controls in both interstitial and bacterial cystitis . In interstitial cystitis mast cells were significantly larger within the detrusor than in the urothelium/submucosa and they appeared to degranulate predominantly within the superficial layers . Differential staining techniques, using long and short toluidine blue stains, failed to reveal statistically significant evidence of mast cell heterogeneity within the bladder wall in interstitial cystitis. J Hosp Infect, 1991 Nov, 19(3), 175 - 80 The effect of surgical theatre head-gear on air bacterial counts; Humphreys H et al.; The wearing of disposable head-gear in operating theatres is currently recommended for scrubbed and non-scrubbed staff . However, there is little evidence of its effectiveness as an infection control measure in casual or non-scrubbed theatre staff . The effect of head-gear on bacterial air counts was studied, using six volunteers, in a sealed room, with and without ventilation . Using a Casella slit sampler and a SAS Air Sampler, air counts ranged from 0.08 to greater than 2.9 colony forming units (cfu) m-3 . The wearing of head-gear was not associated with a reduction in air counts but counts were lower with ventilation . We recommend that non-scrubbed staff no longer wear head-gear as effective ventilation probably counteracts any possible increased bacterial shedding . Scrubbed staff should continue to wear disposable head-gear because of their proximity to the operative field. Mol Gen Mikrobiol Virusol, 1991 Nov, (11), 17 - 20 {Obtaining transgenic Solanum tuberosum potato plants with active bacterial genes xyl and T-cyt effecting the phytohormone balance}; Krasheninnikova LV et al.; The genetic constructions based on integrative vector pGV3850 were used to introduce bacterial genes xyl and T-cyt into potato cells . The transformation was carried out using the leaf-disc method with modifications . A special system for obtaining regenerants from explants of potato in vitro plants or calli has been designed that permitted the selection of transgenic shoots . The presence of the genes in potato genome has been proved by testing the NPTII and glucoisomerase activities . The transgenic plants expressing T-cyt gene differed from the wild type in sharp decrease of the apical dominance. Proc Natl Acad Sci U S A, 1991 Oct 15, 88(20), 8885 - 9 Direct observation of vibrational coherence in bacterial reaction centers using femtosecond absorption spectroscopy; Vos MH et al.; It is shown that vibrational coherence modulates the femtosecond kinetics of stimulated emission and absorption of reaction centers of purple bacteria . In the DLL mutant of Rhodobacter capsulatus, which lacks the bacteriopheophytin electron acceptor, oscillations with periods of approximately 500 fs and possibly also of approximately 2 ps were observed, which are associated with formation of the excited state . The kinetics, which reflect primary processes in Rhodobacter sphaeroides R-26, were modulated by oscillations with a period of approximately 700 fs at 796 nm and approximately 2 ps at 930 nm . In the latter case, at 930 nm, where the stimulated emission of the excited state, P*, is probed, oscillations could only be resolved when a sufficiently narrow (10 nm) and concomitantly long pump pulse was used . This may indicate that the potential energy surface of the excited state is anharmonic or that low-frequency oscillations are masked when higher frequency modes are also coherently excited, or both . The possibility is discussed that the primary charge separation may be a coherent and adiabatic process coupled to low-frequency vibrational modes. J Biol Chem, 1991 Oct 15, 266(29), 19688 - 96 Cloning, bacterial expression, purification, and characterization of the cytoplasmic domain of rat LAR, a receptor-like protein tyrosine phosphatase; Pot DA et al.; This report describes the cloning and characterization of rat leukocyte common antigen-related protein (rLAR), a receptor-like protein tyrosine phosphatase (PTPase) . The recombinant cytoplasmic PTPase domain was expressed at high levels in bacteria and purified to homogeneity . Kinetic properties of the PTPase were examined along with potential modulators of PTPase activity . Several sulfhydryl-directed reagents were effective inhibitors, and a surprising distinction between iodoacetate and iodoacetamide was observed . The latter compound was an extremely poor inhibitor when compared to iodoacetate, suggesting that iodoacetate may interact selectively with a positive charge at or near the active site of the enzyme . Site-directed mutants were made at 4 highly conserved cysteine residues found at positions 1434, 1522, 1723, and 1813 within the protein . The Cys-1522/Ser mutation resulted in a 99% loss of enzymatic activity of the pure protein . This observation is consistent with greater than 99% of the PTPase activity being found in the first domain of the PTPase and demonstrates the critical importance of this cysteine residue in catalysis . The recombinant C1522S mutant phosphatase could also be phosphorylated in vitro by protein kinase C and p43v-abl tyrosine kinase . When pure recombinant PTPase was mixed with 32P-labeled tyrosine substrate and then rapidly denatured, a 32P-labeled enzyme intermediate could be trapped and visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The catalytically inactive C1522S mutant did not form the phosphoenzyme intermediate. Am Rev Respir Dis, 1991 Oct, 144(4), 917 - 22 Mode of presentation and diagnosis of bacterial pneumonia in human immunodeficiency virus-infected patients; Magnenat JL et al.; Bacterial pneumonia (BP) has recently been reported to be more frequent in human immunodeficiency virus (HIV)-infected patients than in normal hosts . This study reviews the clinical and radiologic manifestations of BP in 132 consecutive pulmonary episodes over a 15-month period . BP was defined on a clinical basis as a pulmonary infiltrate accompanied by fever and improving in a few days with conventional antibiotics (trimethoprim-sulfamethoxazole excluded) . In patients undergoing bronchoscopy (97 procedures), semiquantitative cultures and cell differentials of bronchoalveolar lavage (BAL) were performed, in addition to conventional staining and cultures for opportunistic infections . BP were frequent (45%), and the usual community-acquired pathogens were found . The radiologic manifestations of BP were often unusual, however, and 47% were indistinguishable from the typical appearance of Pneumocystis carinii pneumonia . BAL cultures had a sensitivity of 83 or 23%, depending on whether antibiotics were administered before bronchoscopy, using a cutoff value of greater than or equal to 10(4) bacteria/ml . The specificity of BAL culture was of 80.5% if patients with P . carinii pneumonia were taken as a control group . We conclude that BP is frequently encountered in HIV-infected patients . The clinical and radiologic presentation of BP may be indistinguishable from that of opportunistic infections . Semiquantitative cultures of BAL appear a valuable diagnostic tool to avoid unnecessary invasive diagnostic procedures or treatments. Am J Physiol, 1991 Oct, 261(4 Pt 2), H1190 - 6 Splanchnic vasoconstriction and bacterial translocation after thermal injury; Jones WG 2nd et al.; Gut barrier failure and bacterial translocation (BT) after thermal injury may result from splanchnic vasoconstriction and intestinal ischemia . The role of the renin-angiotensin system in intestinal blood flow and BT after thermal injury was studied by pretreatment with the angiotensin-converting enzyme (ACE) inhibitor enalapril in Wistar rats before sham or 30% scald burn . Adequacy of ACE inhibition was documented by the absence of a hypertensive response to angiotensin I, and intestinal blood flow was determined using 51Cr-labeled microspheres . Small bowel blood flow was decreased by 46% at 4-h postburn (P less than 0.05) in untreated burned animals despite maintenance of normal cardiac index but returned to baseline levels by 24 h after injury . Enalapril pretreatment resulted in maintenance of small bowel blood flow after thermal injury and was associated with a significantly reduced incidence of BT (20% vs . 75% in untreated burned animals, P less than 0.01) . These findings further implicate intestinal ischemia in the etiology of gut barrier dysfunction after thermal injury, mediated in part by activation of the renin-angiotensin system. Am J Ophthalmol, 1991 Oct, 112(4 Suppl), 2S - 9S A review of bacterial keratitis and bacterial conjunctivitis; Limberg MB; Bacterial keratitis is an opportunistic infection of the avascular corneal stroma, initiated by a breakdown of the epithelial barrier . Corneal abrasion with infected material, extended-wear contact lenses, eyelid disease, and disorders of the ocular surface are predominant contributing factors . Acute bacterial conjunctivitis occurs when sufficient bacteria are introduced into the fornices to overwhelm normal bacteriostatic and flushing mechanisms . Chronic conjunctivitis is associated with a constant adnexal source of bacteria and resolves when the source is successfully treated . Rapid diagnosis and treatment of bacterial keratitis are essential to limit stromal scarring and minimize visual loss. Transfusion, 1991 Oct, 31(8), 710 - 2 Effect of blood transfusion on survival in a mouse bacterial peritonitis model; Goldman M et al.; Allogeneic blood transfusions can result in alloimmunization or immunosuppression . A previous study demonstrated a deleterious effect of allogeneic blood transfusion on tumor growth in mice that was dependent, in part, on the dose of tumor cells with which the host animal was inoculated . The current study examined the effect of a similar allogeneic blood transfusion protocol on survival in a mouse bacterial peritonitis model . C57Bl/6J mice were transfused with 0.2 mL of heparinized fresh whole blood from C57Bl/6J (syngeneic) or Balb/c (allogeneic) mice . Transfusions were given on Days -10 and -7 . On Day 0, mice were injected intraperitoneally with 10(7) Escherichia coli . Survival at Day 7 was 61 percent in the allogeneic blood transfusion group and 55 percent in the syngeneic blood transfusion group (p = 0.52) . Experiments using different strains of mice, different transfusion protocols, and different doses of bacteria also failed to demonstrate an effect of allogeneic blood transfusion on survival . The results demonstrate that blood transfusion does not influence survival after a septic challenge with bacteria . The data obtained in the present study, together with those obtained in the tumor model, suggest that the mechanisms by which the allogeneic blood transfusion impedes host defense against bacterial infections is different from the mechanisms involved in tumor growth. Surgery, 1991 Oct, 110(4), 685 - 8; discussion 688-90 The effect of hypertonic saline resuscitation on bacterial translocation after hemorrhagic shock in rats; Reed LL et al.; Translocation of enteric bacteria occurs in rats after hemorrhagic shock . A proposed mechanism involves intestinal mucosal injury by hypoperfusion . Recent work suggests that moderate hypovolemia causes gut arteriolar constriction, which is ameliorated by hypertonic saline resuscitation . Bacterial translocation should, therefore, be reduced when hypertonic saline (HS) is used as the resuscitative fluid . Seventy-eight Sprague-Dawley rats were anesthetized and subjected to 30 minutes of hemorrhagic shock (systolic blood pressure 30 to 50 mm Hg) through a modified Wigger's model . Resuscitation was performed with either shed blood (B), 3% HS + 1/2B (1:1), or with 7.5% HS + 1/2B (1:1) . Spleen, liver, and mesenteric lymph nodes were sent for quantitative culture 24 hours later . Translocation occurred if enteric organisms were cultured from at least one organ . Statistical analysis used the Fisher exact test . Compared to autotransfusion, hemodilutional resuscitation from hemorrhagic shock with hypertonic saline resulted in a significant reduction in bacterial translocation (p values were 0.03 and 0.04 for 3% and 7.5% hypertonic saline, respectively) . The reduction in translocation after hypertonic saline resuscitation may be the consequence of microcirculatory alterations preventing gut hypoperfusion. J Bacteriol, 1991 Oct, 173(19), 5999 - 6008 The Escherichia coli htrP gene product is essential for bacterial growth at high temperatures: mapping, cloning, sequencing, and transcriptional regulation of htrP; Raina S et al.; We identified and characterized a new Escherichia coli gene, htrP . The htrP gene was identified because its insertional inactivation by the Tn10 transposon results in the inability of E . coli to form colonies at temperatures above 37 degrees C and a slow growth phenotype at 30 degrees C . The htrP gene was cloned and mapped to 66.3 min on the E . coli genetic map, 4 kbp clockwise from the tolC gene . The htrP gene was sequenced and shown to code for an acidic, 27,471-Da polypeptide and to be transcribed counterclockwise with respect to the genetic map . The predicted HtrP protein has two potential transmembrane segments and shares an identity of 64.4% over a length of 210 amino acids with the LuxH protein . Despite the fact that the htrP gene is essential for E . coli growth exclusively at high temperatures, the levels of htrP-specific transcripts decrease with increasing temperature. Infect Immun, 1991 Oct, 59(10), 3646 - 50 Tumor necrosis factor and interleukin-1 activities in free lung cells after single and repeated inhalation of bacterial endotoxin; de Rochemonteix-Galve B et al.; Bacterial endotoxins (lipopolysaccharides), important components of many organic dusts, are known to induce macrophages to produce the inflammatory mediators interleukin-1 (IL-1) and tumor necrosis factor alpha (TNF-alpha) . To investigate the role of these mediators in the early inflammatory responses in the lung, guinea pigs were exposed to an aerosol of bacterial endotoxin . A bronchoalveolar lavage (BAL) was then performed, and TNF-alpha and IL-1 in lysed BAL cells and in the supernatants from BAL cell cultures were studied . The effect of single and repeated LPS inhalation exposures on the activities of TNF and IL-1 was studied, as was the effect of LPS added to the cell culture medium . A single inhalation exposure to LPS caused an increase in the TNF-alpha and IL-1 activities in cell lysate and in the cell culture supernatant . After a second inhalation exposure, cell-associated and extracellular TNF-alpha activity could not be detected, whereas IL-1 activity was markedly enhanced . IL-1 activity was increased when LPS was added to the cell culture medium with or without a prior inhalation exposure . In contrast, TNF-alpha activity was not affected after a second exposure. Rev Invest Clin, 1991 Oct-Dec, 43(4), 329 - 33 {Bacterial count as an infection prognostic factor in the delayed primary closure of abdominal surgical wounds}; Herrera Hernandez MF et al.; STUDY OBJECTIVE . To determine the prognostic value of the soft-tissue biopsy to handle contaminated or infected surgical wounds with delayed primary closure . DESIGN . Comparative, prolective, blind and observational . PLACE . Referral tertiary care center . PATIENTS . We included 70 patients distributed in two groups: Group I: 50 patients who underwent contaminated or infected abdominal surgery; and Group II: 20 patients who underwent clean or clean contaminated abdominal surgery and developed surgical wound abscess . INTERVENTIONS . Daily cleaning and water irrigation and periodic debridement were done in all the cases; after five days (excluding infection), two tissue biopsies for quantitative culture and wound closure were performed . MEASUREMENTS AND MAIN RESULTS . In Group I, nine of 26 patients (35%) with positive culture (greater than 10(5) colony forming units/g) developed infection and none of 24 with negative culture (p less than 0.01, Fisher); the negative predictive value (nPV) was 100% and the positive predictive value (pPV) was 35% . In Group II, seven of 12 patients with positive culture and one of eight with negative culture developed infection (p less than 0.05); the nPV was 87% and the pPV was 58% . CONCLUSIONS . The biopsy of soft-tissue seems to be a helpful tool for the treatment and prognosis of infected or contaminated surgical wounds. Vrach Delo, 1991 Oct, (10), 78 - 81 {The regulatory function of the hemato-encephalic barrier in bacterial meningoencephalitis}; Iarosh OO; The author studied the permeation of the blood brain barrier to metals in 70 patients depending on the severity and terms of meningoencephalitis . It is shown that use of the probability-statistical analysis of the quantitative content of metals in in the blood and cerebrospinal fluid allows to fix the regimen of the blood brain barrier work from initial manifestations of functional insufficiency up to disorders of regulatory mechanisms and their metabolism. Mikrobiyol Bul, 1991 Oct, 25(4), 387 - 90 {Bacterial classification}; Asan A; In this paper, the essential points in classification of bacteria were given . In addition, a short history of the classification and the medical importance of the bacteria were listed. Arch Tierernahr, 1991 Oct, 41(7-8), 703 - 16 {Determination of bacterial N portions in feces and differently collected ileum chymus from swine}; Wunsche J et al.; In faeces and ileal digesta samples of 31 intact (INT) as well as 73 surgically differently prepared pigs bacterial fractionations and 2.6-diaminopimelic acid (DAP) estimations were carried out in order to calculate the bacterial N proportion in faeces N and digesta N after feeding various diets . Because of a high individual variability and the analytical variation width of the DAP/N-ratios no distinct influences of the fed diets could be found . The average DAP/N-values in the faeces (0.224) of INT pigs ranged in the same magnitude as in the digesta (0.0272) of ileorectostomized (IRA) pigs with open colon descendens (IRAo), where a digesta backflow is possible . Distinct lower DAP/N-ratios (0.0125 resp . 0.0043), however, were found in the digesta of pigs with ileo-caecal cannulae (IZB) or IRA pigs with closed colon (IRAg) . On the base of various premises (N of the "bacterial fraction" C is only bacterial N; the DAP found in fraction A originates from intestinal bacteria adhering to feed particles) conducted calculations of the bacterial N proportions (in per cent of total N) led to the following data: Faeces of INT pigs: 43.0 .. . 68.2 vs . 69.6 .. . 89.0; digesta of operated pigs (except protein free diet) IRAo: 22.3 .. . 57.0 vs . 46.2 .. . 73.8; IZB: 17.0 .. . 35.7 vs . 25.2 .. . 53.6; IRAg (only 3 pigs): 23.6 vs . 24.2 . The proportion of bacterial N in the digesta N of protein free fed IRAo pigs was 22.0 vs . 22.6%. J Gen Microbiol, 1991 Oct, 137 ( Pt 10), 2313 - 9 Transcription of a stem/loop region of a tumour-amplified sequence induces bacterial aggregation; Heighway J et al.; A sequence of human DNA, amplified in a lung tumour, was shown to induce expression-dependent effects on Escherichia coli phenotype . Previous studies have demonstrated the induction of abnormal plasmid supercoiling and bacterial aggregation in host strains harbouring various constructs encoding this region . Subcloning identified a short stretch of 50 bp crucial for these effects . This study details further characterization of the active sequence . Computer analysis of the region predicted the formation of a stem/loop structure in transcribed RNA . Transcription, in the absence of translation, of a 22 bp sequence comprising this structure was sufficient to induce bacterial aggregation . Site-directed and random mutagenesis of the sub-region were carried out in order to identify critical factors in the induction of this phenotypic shift . It was found that changes in the loop sequence modulated activity, and mutations increasing predicted stem stability produced more active constructs . The wild-type sequence induced high level aggregation in only a limited number of strains but using the mutagenesis data, a sequence was synthesized that induced high level aggregation in most E . coli strains tested. Mol Biochem Parasitol, 1991 Oct, 48(2), 185 - 98 An 85-kilodalton surface antigen gene family of Trypanosoma cruzi encodes polypeptides homologous to bacterial neuraminidases; Takle GB et al.; We have determined the sequence of a cDNA (Tt34c1) encoding a Trypanosoma cruzi trypomastigote stage-specific 85-kDa surface glycoprotein (gp85) . Within the peptide sequence of Tt34c1 are two 8-amino acid motifs, Ser-X-Asp-X-Gly-X-Thr-Trp, that are characteristic of bacterial neuraminidases . Analysis of the Tt34c1 sequence predicts the presence of an amino-terminal signal sequence and a hydrophobic carboxy-terminus that is probably replaced by a glycosyl phosphatidylinositol membrane anchor . Gp85 is encoded by an extensive multigene family that is distributed throughout the genome and can be divided into subsets on the basis of oligonucleotide hybridisation patterns . By sequencing products of polymerase chain reaction (PCR) amplification of the 5' end of trypomastigote gp85 mRNA we show that multiple copies of the gene family are transcribed simultaneously in a trypanosome population . Comparison of the sequence of the PCR clones and another gp85 cDNA showed a highly conserved region 5' of the first methionine extending 180 nt into the coding sequence . Insertions and point mutations were observable outside these homologous sequences demonstrating the variant nature of the gp85 mRNAs. Biochem Biophys Res Commun, 1991 Sep 16, 179(2), 872 - 9 Bacterial toxins induce heat shock proteins in human neutrophils; Hensler T et al.; We studied the influence of different bacterial toxins (alveolysin; toxic shock syndrome toxin 1, TSST-1 and erythrogenic toxin A, ETA) on the expression of heat shock proteins (hsps) in isolated human polymorphonuclear granulocytes (PMNs) . As was shown by Western blotting (anti-hsp72) ETA and TSST-1 were potent inducers of hsps at low toxin concentrations (10 ng/ml) . Alveolysin led to the expression of hsps at hemolytic concentrations (1 HU; 700 ng/ml) whereas at subhemolytic concentrations (7 ng/ml) no heat shock response was observed . The induction of heat shock proteins was also accompanied by increased mRNA levels for hsp70 as was determined by PCR-analysis. J Biol Chem, 1991 Sep 15, 266(26), 16969 - 72 Identification of an amino acid-regulated mRNA from rat liver as the mammalian equivalent of bacterial ribosomal protein L22; Laine RO et al.; Amino acid deprivation of rat hepatoma cells induced the levels of a 612-base pair mRNA termed ASI (Shay, N . F., Nick, H . S., and Kilberg, M . S . (1990) J . Biol . Chem . 265, 17844-17848) . The ASI mRNA was present at levels equal to or greater than actin in every rat tissue tested . The corresponding full-length cDNA was cloned, and the present report demonstrates that the deduced 184-residue amino acid sequence shares greater than 30% identity to a number of bacterial and chloroplast L22 ribosomal proteins, including those from Escherichia coli and Halobacterium halobium . A monospecific anti-peptide antibody was produced that upon immunochemical analysis of subcellular fractions of rat liver recognized a band in the microsomal fraction and, more specifically, reacted with a single polypeptide in the ribosomal large subunit fraction . The antibody did not react with any proteins of the mitochondrial large subunit, but did recognize a protein in human liver homogenate at the same relative mobility (23 kDa) as that observed for rat liver. J Immunol, 1991 Sep 15, 147(6), 1759 - 64 Stimulation of in vitro murine lymphocyte proliferation by bacterial DNA; Messina JP et al.; Although DNA is generally considered to be a poor immunogen, recent evidence suggests that DNA from various species differ in their immunologic activity and that bacterial DNA, unlike mammalian DNA, can induce significant antibody responses in mice . To explore further the immunologic activities of bacterial DNA, its ability to stimulate in vitro proliferation of murine lymphocytes was tested . The stimulation of lymphocytes with highly purified ssDNA from Escherichia coli resulted in a dose-dependent response that was maximal at 48 h . Several lines of evidence indicate that DNA, rather than endotoxin contamination, induced this response: 1) LPS at doses equivalent to those detected in the DNA preparation caused significantly less proliferation than the DNA; 2) the response to DNA was insensitive to polymyxin B; 3) pretreatment of DNA with DNase completely abrogated the response; and 4) DNA induced the proliferation of cells from endotoxin-resistant C3H/HeJ mice . Furthermore, although DNA from three different bacterial species induced proliferation, mammalian DNA from three species were nonmitogenic . Depletion of T cells from lymphocytes did not reduce proliferation, suggesting that bacterial DNA directly triggered B cell proliferation . These studies provide further evidence that DNA are not uniform in their immunologic activities likely because of their content of nonconserved structural determinants. Int J STD AIDS, 1991 Sep-Oct, 2(5), 362 - 4 Is bacterial vaginosis a sexually transmitted disease? Larsson PG, Platz-Christensen JJ, Sundstrom E. The sexual behaviour of 400 women with and 400 women without bacterial vaginosis from a health screening programme was investigated by a questionnaire to see if women with bacterial vaginosis had similar behaviours to women considered at risk for STDs . These patients were randomly selected from computer records . Eight hundred questionnaires were distributed and 641 women answered . The age of first sexual intercourse was lower among women with bacterial vaginosis (17.8 years) than those without (18.6 years; P less than 0.001) . Number of lifetime sexual partners was higher among women with bacterial vaginosis (P less than 0.001) . Women with bacterial vaginosis smoked or had smoked more often . These results indicate that women with bacterial vaginosis have similar sexual behaviour to those at risk for STDs. J Emerg Med, 1991 Sep-Oct, 9(5), 317 - 21 Prevention of oral bacterial flora transmission by using mouth-to-mask ventilation during CPR; Cydulka RK et al.; The Emergency Cardiac Care Committee of the American Heart Association has recently recommended utilizing protective barrier precautions during CPR (1,2) . We assessed 17 mask and faceshield resuscitation devices for adequacy of barrier protection . Eight of the devices were faceshields (CPR Microshield, Hygenic, MedCare Mask, Resusci, Samaritan, Sealeasy, Portex); 8 were mask devices (Laerdal, Dyna Med, MTM Emergency Lung Ventilator, MTM Emergency Resuscitator, Res-Q-Flo, Rightway Mouth-to-Mask Resuscitation, Trufit), and one of the devices did not meet the criteria for either faceshield or mask (Lifesaver) . All masks were disinfected, applied to the investigator's face as directed by the manufacturers' instructions, and then cultured for oral aerobic bacterial flora on the rescuer side . No mask devices cultured positive for oral aerobic bacterial flora, while 6 of 8 faceshield devices cultured positive for oral aerobic bacterial flora (P less than 0.007) . The CPR Microshield and the Portex faceshield were the only devices that did not develop a positive culture . We conclude that all ventilation devices with a one-way valve, except the Sealeasy device, provide adequate barrier type protection from oral aerobic bacterial flora when simulating mouth-to-barrier type protection when performing mouth-to-mouth ventilation. Clin Obstet Gynecol, 1991 Sep, 34(3), 582 - 6 Bacterial vaginitis; Faro S; Bacterial vaginitis is responsible for approximately 10 million office visits per year . This condition may disrupt marriages and cause psychologic stress that may be reflected in the individual's work and social life . Bacterial vaginitis also has been thought to lead to postpartum endometritis, pelvic cellulitis, pelvic inflammatory disease, and chorioamnionitis. Clin Exp Immunol, 1991 Sep, 85(3), 510 - 4 Analysis of the molecular mimicry between HLA-B27 and a bacterial OmpA protein using synthetic peptides; Yu DT et al.; In spite of a lack of sequence 'homology' between HLA-B27 and the bacterial OmpA outer membrane proteins, they both react with the Ye-2 monoclonal anti-HLA-B27 antibody . The Ye-2 antibody also reacted positively in ELISA with a synthetic peptide derived from the segment spanning residues 63-84 of B*2705 . The critical peptide residues were determined by testing first with overlapping peptides, followed by a replacement set made according to the determined epitope . The results were compared with those with overlapping eight mers made to span a carboxyl fragment of the Escherichia coli OmpA protein . They indicate the reason why Ye-2 reacts with both sets of peptides is because it has a preference for polymers of arginine. Am J Obstet Gynecol, 1991 Sep, 165(3), 737 - 42 A new proline aminopeptidase assay for diagnosis of bacterial vaginosis; Schoonmaker JN et al.; Bacterial vaginosis is one of the most common occurring vaginal conditions among women of reproductive age . A rapid and reliable laboratory test for diagnosis of bacterial vaginosis would be helpful in the clinical detection of this disease . Elevated proline aminopeptidase activity has been identified as a reliable marker enzyme for bacterial vaginosis . A proline aminopeptidase assay has been shown to predict accurately women with a clinical diagnosis of bacterial vaginosis . However, this assay has significant practical disadvantages, the most notable of which is the production of a carcinogenic end product, alpha-naphthylamine . We have developed a modified assay for this bacterial vaginosis marker enzyme with L-proline p-nitroanilide, a substrate that does not yield a carcinogenic end-product . The new proline aminopeptidase assay is a one-step test that is analyzed colorimetrically with microsomal leucine aminopeptidase used as a standard enzyme (linear from 3 to 125 mU per well) . We have determined the activity of proline aminopeptidase in vaginal wet preparations from 57 patients with both assay methods . In addition, vaginal smears were examined with Gram's stain and analyzed for bacterial vaginosis with the Spiegel method . When compared with the Spiegel method, the two proline aminopeptidase assay methods were similar with respect to assay sensitivity (93%), specificity (91% to 93%), and the predictive value of a positive result (78% to 82%) or a negative result (97% to 98%) . Vaginal wash samples also were assessed for proline aminopeptidase activity . Values for samples identified as bacterial vaginosis positive were significantly different (p less than 0.0001) from those that were negative according to the Spiegel analysis of Gram's stain: negative results, 66 +/- 41 mU/ml; positive results, 704 +/- 145 mU/ml . These findings indicate that this improved proline aminopeptidase assay will offer a rapid, sensitive, and objective laboratory method for the diagnosis of bacterial vaginosis. Chest, 1991 Sep, 100(3), 644 - 8 Bacterial ribosomal immunostimulants prime alveolar macrophages in vivo to produce interleukin 1 in vitro; Pujol JL et al.; Alveolar macrophages (AMs) may play a key role in human respiratory immune defenses, partially by synthesizing and releasing interleukin 1 (IL = 1) . D53 (Ribomunyl), a composite bacterial ribosomal immunostimulant, has been recognized as an efficient prevention of respiratory tract infections . In vitro, D53 enhances the IL-1 production by mouse spleen adherent cells . A thymocyte proliferative response assay was used to evaluate the in vitro IL-1 production by AMs in healthy subjects who received D53 immunostimulant . Twelve nonsmoking healthy subjects took part in a prospective double-blind placebo control study . On day 1, a first bronchoalveolar lavage (BAL) was performed to assess IL-1 production by unstimulated and lipopolysaccharide (LPS) stimulated AM . Then, subjects were randomized to receive D53 (n = 6) or its placebo (n = 6) by both oral and subcutaneous injection routes from day 1 to day 15 . On day 15, a second BAL was done and AM IL-1 production was again tested . IL-1 production on day 15 did not significantly differ from day 1 in both D53-treated and placebo groups either when AMs were unstimulated or were stimulated with concentrations of LPS resulting in maximal IL-1 production . However, in the D53-treated group, but not in the placebo group, IL-1 production induced by low LPS concentration (5 mg/L) was significantly higher (mean +/- SEM: 1,238 +/- 287 U/10(6) AM) on day 15 in comparison with day 1 (577 +/- 113 U/10(6) AM; p less than 0.05, Wilcoxon W test) and in comparison with the control group (day 15 IL-1 production induced by 5 mg/L LPS, 758 +/- 175 U/10(6) AM; p less than 0.05, Mann-Whitney U test) . Moreover, in the D53-treated group, the optimal LPS concentration (ie, LPS concentration that induced maximal IL-1 production) was significantly lower on day 15 (mean +/- SD: 11 +/- 7 mg/L) than on day 1 (16 +/- 7 mg/L; p less than 0.05 Wilcoxon W test) . We conclude that D53 immunostimulant in vivo primes AM to produce IL-1 following low LPS concentration stimulation . This may partially explain the protective effect of D53 immunostimulant against respiratory tract infection. Am J Physiol, 1991 Sep, 261(3 Pt 2), R659 - 64 Differential feeding responses to bacterial lipopolysaccharide and muramyl dipeptide; Langhans W et al.; The present study was designed to test whether a tolerance to the hypophagic effects of bacterial lipopolysaccharide (LPS) and muramyl dipeptide (MDP) develops with repeated injections in rats and whether a relationship between the hypophagic effects of both compounds exists . Only the first of three subsequent intraperitoneal injection of LPS (100 micrograms/kg body wt each), given every 2nd day, led to a significant reduction of food intake . In contrast, MDP (1.6 mg/kg body wt) did not lose its hypophagic effect with four subsequent intraperitoneal injections . Furthermore, the LPS tolerance did not alter the hypophagic response to subsequently injected MDP . Likewise, MDP pretreatment did not alter the hypophagic response to LPS . Doses of MDP and LPS that were individually below the threshold for a reliable reduction of food intake (0.4 mg MDP/kg body wt + 25 micrograms LPS/kg body wt) reduced food intake synergistically when injected together . The hypophagia produced by combined injections of MDP plus LPS slowly diminished with repeated injections . The results indicate that separate but interacting mechanisms are involved in the feeding responses to MDP and LPS . The observed synergism between MDP and LPS suggests a synergistic role of bacterial muramyl peptides and LPS in the anorexia during bacterial infections. J Surg Res, 1991 Sep, 51(3), 267 - 70 Liver bacterial clearance following hepatic artery ligation and portacaval shunt; Katz S et al.; The reticuloendothelial system (RES) plays an important role in removing bacteria, endotoxins, and immune complexes from the circulation . Hepatic phagocytosis accounts for more than 80% of RES function . The dual hepatic blood supply (hepatic artery/portal vein) may be altered by pathologic states and surgical procedures . This study evaluates and compares the effect of hepatic artery ligation and portacaval shunt on hepatic trapping of viable Escherichia coli . Thirty rats were placed in three groups: Group I was composed of sham operated controls; Group II underwent end-to-side portacaval shunt (PCS); and in Group III, hepatic artery ligation (HAL) was performed . At 2 weeks following the operation 10(9) 35S-radiolabeled viable E . coli were injected via the tail vein . At 10 min, bacterial distribution in the different organs was determined . Tissue samples were processed for liquid scintillation counting . The final distribution of bacteria was calculated from the input specific activity (dpm/bacteria) and expressed as the mean percentage of injected viable E . coli per gram of tissue and per organ weight . There was a significant decrease of bacterial trapping by the liver in rats following PCS (Group II), 45.0 +/- 10.4% vs controls 77.1 +/- 3.73% (P less than 0.005) . This was partially compensated for by a significant increase of bacterial trapping by the lung . The decreased clearance in PCS rats is due to a reduction in liver mass compared to that in controls . Bacterial localization in HAL (Group III) rats was similar to that in controls . These data show that PCS decreases hepatic clearance and increases pulmonary localization of viable E . coli . This phagocytic dysfunction may contribute to increased susceptibility to infection following portacaval shunt. J Immunol, 1991 Sep 1, 147(5), 1463 - 9 B cell triggering by bacterial lipopeptide involves both translocation and activation of the membrane-bound form of protein kinase C; Bosca L et al.; B cell activation by the lipopeptide trispalmitoyl-cysteinyl-alanyl-glycine (TPP), the biologically active moiety of bacterial lipoprotein, results in protein kinase C (PKC) translocation from the cytosol to the plasma membrane, as well as a significant increase in the activity of membrane-associated PKC that can be observed by in vitro incubation with TPP of partially purified PKC at calcium concentrations in the range of those prevailing in unstimulated B cells . TPP does not affect either the phosphoinositide turnover or the cytosolic-free calcium concentration, but promotes an increase in the intracellular pH that can be blocked by the PKC-inhibitors staurosporine or H-7 . Moreover, incubation of B cells with staurosporine suppressed the proliferative response promoted by TPP at a half-maximum effective dose of 16 nM . Activation by TPP of PKC isoenzymes resolved after hydroxylapatite chromatography revealed that the resulting beta I/beta II isoenzyme was more sensitive than the alpha isoenzyme . These results suggest that TPP might mediate B cell activation via interaction with the membrane-associated fraction of PKC. Infect Immun, 1991 Sep, 59(9), 2994 - 8 Differential sensitivity of CD8+ suppressor and cytotoxic T lymphocyte activity to bacterial monophosphoryl lipid A; Esquivel F et al.; Treatment with a preparation of monophosphoryl lipid A, known to be capable of abolishing the expression of CD8+ suppressor T cell activity generated during the antibody response to type III pneumococcal polysaccharide (SSS-III), was found to have no adverse effect upon either induction or expression of CD8+ cytotoxic T lymphocyte activity specific for influenza A virus antigens . This suggests that suppressor T cells and cytotoxic T lymphocytes represent functionally distinct subsets of CD8+ T cells which can be differentiated on the basis of their sensitivities to inactivation by monophosphoryl lipid A. Int J Dev Biol, 1991 Sep, 35(3), 239 - 49 The influence of mouse sera, regenerating liver extracts and bacterial products on the abilities of different cells in vitro; Zarkovic N et al.; In the complexity of host tumor relations, the regeneration of the tissue in which the tumor is growing, or in some other tissue in the organism, could influence the maturation of tumor cells, i.e . tumor reversion . Clinical observations and experiments on plants, lower animals, or animal embryos, performed by several authors, and our results on the influence of regenerating mouse liver on the abilities of tumor transplanted there or elsewhere in the organism led us to study the in vitro growth of different cells or bacteria exposed to the extracts of normal or regenerating liver and/or sera from these animals . Further, sterile used bacterial media were added to bacterial or cell cultures, respectively . Depending on the model, liver extracts-particularly extracts and sera from mice with regenerating liver-were shown to inhibit radioactive thymidine incorporation in the cells . In these experiments, the number of bacteria or cells per culture was lower than in otherwise treated corresponding cultures . Further, used sterile media of bacterial cultures stimulated the growth of bacteria but inhibited thymidine incorporation into fibrosarcoma cells in vitro . Whether this means that one or several common regulators exist in nature appears as an intriguing, but still completely open question . The idea of controlling tumor growth by using such regulatory growth factors seems very provocative. Nutrition, 1991 Sep-Oct, 7(5), 358 - 63 Effect of three liquid diets on cecal bacterial flora and bacterial translocation in mice; Wells CL et al.; Separate groups of mice were fed either standard rodent chow or one of three liquid diets (Impact, Isosource HN, Fibersource HN) for 14 days to determine the effects of these liquid diets on the cecal bacterial flora and on the incidence of bacterial translocation to the mesenteric lymph nodes . Liquid Isosource and liquid Fibersource had no noticeable effect on either the cecal bacterial flora or the incidence of bacterial translocation . Liquid Impact was associated with cecal bacterial overgrowth but had no effect on the incidence of bacterial translocation . Each of the liquid diets was then lyophilized and similarly fed to mice; none of the lyophilized diets had a noticeable effect on the cecal bacterial flora or the incidence of bacterial translocation . To test these liquid and lyophilized diets in stressed mice, separate groups of mice were again fed the various diets for 14 days but given parenteral Escherichia coli lipopolysaccharide (LPS) 24 h before being killed . None of the liquid or lyophilized diets had a noticeable effect on the cecal bacterial overgrowth noticed in LPS-treated chow-fed mice . Compared with chow-fed mice, the typically elevated LPS-induced bacterial translocation was even more elevated in mice fed liquid diet but was somewhat decreased in mice fed lyophilized diet . Thus, the modulatory effects of these liquid diets on the cecal flora and the incidence of bacterial translocation appeared to depend on the composition of the diet and on prior treatment with parenteral LPS. G Ital Cardiol, 1991 Sep, 21(9), 1011 - 5 { Bacterial endocarditis of the pulmonary valve damaged by thoracic radiotherapy (in Hodgkin's disease)}; Conte MR et al.; It is well known that radiation therapy to the anterior mediastinum may induce lesions of all cardiac structures . The pericardium is most frequently involved, but atrioventricular conduction disorders, cardiomyopathy, coronary stenosis may also be produced . Aortic, mitral and tricuspid lesions have been described . However, clinical evidence of pulmonic valve involvement has not been reported . Only at necropsy has fibrotic thickening of the pulmonic cusps occasionally been found . We report a case of infective endocarditis of the pulmonic valve in a 53-year-old patient who had undergone thoracic radiation therapy for Hodgkin's disease 31 years previously . Four years prior to the endocarditis he had also been submitted to myocardial revascularisation for critical lesions of the left main and right coronary ostia, and to aortic valve replacement because of stenosis and insufficiency . At that time, the pulmonic valve was fibrotic on echo examination . It is noteworthy that, of all the cardiac valves, the infective process involved only the pulmonic one, which is seldom the target of an infection . To our knowledge this is the first case of bacterial endocarditis of a heart valve that had been previously damaged by radiation therapy. Photochem Photobiol, 1991 Sep, 54(3), 473 - 6 A new reducing agent of flavins and its application to the assay of bacterial luciferase; Lei BF et al.; Copper(I) plus ethylenediaminetetraacetic acid, Cu(I)-EDTA, reduces flavins . Using Cu(I)-EDTA reduction of riboflavin 5'-phosphate, a new assay method for bacterial luciferase is established . In previous assay methods, flavin and aldehyde substrates for luciferase have been routinely added to luciferase at different times; the new assay permits aldehyde and Cu(I)-reduced flavin to be injected simultaneously into air-saturated buffer containing luciferase . In the new assay method using Cu(I), bovine serum albumin is not required, the initial emission intensity is more than twice and the overall yield is about twice of that in the so-called standard assay for luciferase. Tubercle, 1991 Sep, 72(3), 190 - 2 Adenosine deaminase levels in cerebrospinal fluid in tuberculosis and bacterial meningitis; Chawla RK et al.; Adenosine deaminase activity was measured in cerebrospinal fluid of patients with confirmed tuberculous and bacterial meningitis . The values were compared with those of control subjects without meningitis . A statistically significant increase in the level of this enzyme was noted in the two types of meningitis, but no definite demarcation in the levels was observed between the two types . Therefore increases in adenosine deaminase activity may not be of such diagnostic significance as reported elsewhere. J Mol Evol, 1991 Sep, 33(3), 241 - 50 Molecular considerations in the evolution of bacterial genes; Lawrence JG et al.; Synonymous and nonsynonymous substitution rates at the loci encoding glyceraldehyde-3-phosphate dehydrogenase (gap) and outer membrane protein 3A (ompA) were examined in 12 species of enteric bacteria . By examining homologous sequences in species of varying degrees of relatedness and of known phylogenetic relationships, we analyzed the patterns of synonymous and nonsynonymous substitutions within and among these genes . Although both loci accumulate synonymous substitutions at reduced rates due to codon usage bias, portions of the gap and ompA reading frames show significant deviation in synonymous substitution rates not attributable to local codon bias . A paucity of synonymous substitutions in portions of the ompA gene may reflect selection for a novel mRNA secondary structure . In addition, these studies allow comparisons of homologous protein-coding sequences (gap) in plants, animals, and bacteria, revealing differences in evolutionary constraints on this glycolytic enzyme in these lineages. J Nurse Midwifery, 1991 Sep-Oct, 36(5), 289 - 96 Bacterial vaginosis and its implication in preterm labor and premature rupture of membranes . A review of the literature; Reynolds HD; Preterm delivery continues to occur in 5% to 10% of all births, with a perinatal mortality rate between 50% and 80% . In recent years, the role of infection with lower genital tract organisms in precipitating preterm labor/delivery and premature rupture of membranes has come under considerable study . This article reviews the mechanisms by which infection may play a role in these problems, with a specific focus on bacterial vaginosis . Clinical management issues are addressed, including screening of prenatal patients, diagnostic criteria, and treatment possibilities. Scand J Prim Health Care, 1991 Sep, 9(3), 197 - 202 Treatment of bacterial vaginosis with metronidazole or pivampicillin; Schmidt H et al.; In a double-blind randomized controlled trial from general practice, we assessed the efficacy of treatment with either metronidazole or pivampicillin in patients fulfilling the diagnostic criteria of bacterial vaginosis . 50 women were treated with metronidazole 500 mg twice daily and 54 with pivampicillin 700 mg twice daily, both for seven days . Evaluated four weeks after the start of treatment, we found a significantly higher cure rate in the metronidazole group (90%) than in the pivampicillin group (69%) (p = 0.01) . Cure was defined as lack of fulfillment of the diagnostic criteria of bacterial vaginosis . Test for confounding according to parity and complaint of vaginal discharge showed no influence . Side-effects were significantly commoner in the pivampicillin group (28%) than in the metronidazole group (8%) . We consider that metronidazole is a potent drug in the treatment of bacterial vaginosis, with pivampicillin as a useful alternative. Nippon Saikingaku Zasshi, 1991 Sep, 46(5), 847 - 54 {Augmentation of activities of peripheral granulocytes and of resistance against bacterial infection after administration of G-CSF into mice}; Shinomiya N et al.; We evaluated the metabolic capability of murine peripheral granulocytes after administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) by quantitative flow cytometric assay for H2O2-dependent oxidative product formation . Intraperitoneal administration of a daily dose of 10 micrograms of rhG-CSF for 5 days induced doubling of the leukocyte population . Differential counting of peripheral leukocytes and scattergram by flow cytometry showed an increased mature granulocyte population . After stimulation with phorbol myristate acetate, the granulocytes of the rhG-CSF-administered mice demonstrated some hyperresponsive population and an increased H2O2 production . The hyperresponsive population showed H2O2 production 4-6 times higher than did normal cells . Granulocytes from the G-CSF-treated mice revealed an augmented phagocytic activity and an increased expression of Mac-1 molecules . Moreover, mice treated with G-CSF showed an enhanced resistance against intravenous infection with a lethal dose of E . coli . Granulocytes showing such markedly increased oxidative metabolism may be a significant component of the host defence to various infective organisms. Eur J Surg, 1991 Sep, 157(9), 539 - 42 Bacterial load and inflammatory reaction in the bowel wall after colonic obstruction . An experimental study in rats; Tornqvist A et al.; To quantify the inflammatory reaction to obstruction, alkaline phosphatase and myeloperoxidase activities were measured in specimens of colonic wall from 60 rats . Twenty rats had undergone laparotomy and band obstruction, 20 laparotomy without obstruction and 20 had no operation . The mean activities of both enzymes were increased proximal but not distal to the obstruction, that of alkaline phosphatase significantly so . Despite increase in the number of bacteria, both proximal and distal to the obstruction, no signs of bacterial invasion of the bowel wall were seen either on histological examination or culture . These findings indicate that mechanical factors may be more important than bacteria in causing the increased inflammatory reaction in bowel wall proximal to an obstruction. Mol Gen Genet, 1991 Sep, 229(1), 27 - 30 Distribution of sequences homologous to the impCAB operon of TP110 among bacterial plasmids of different incompatibility groups; Lodwick D et al.; Mutagenic DNA repair is a function of many naturally occurring plasmids belonging to several different incompatibility groups . A DNA probe corresponding to the impCAB operon of the IncI1 plasmid TP110, which encodes such functions, was used to investigate the distribution of homologous sequences in both related and unrelated plasmids . Southern blotting was used to demonstrate considerable sequence conservation amongst a number of plasmid types, with imp-related sequences being found on plasmids belonging to the I1, I1/B, B and FIV incompatibility groups . However, no homology was detected amongst plasmids of the N and L/M incompatibility groups, many of which carry functionally similar gene clusters . It appears that sequences determining mutagenic repair functions have been largely conserved within any one incompatibility group, but that significant divergent evolution has occurred between groups. J Burn Care Rehabil, 1991 Sep-Oct, 12(5), 454 - 7 Bacterial translocation is prolonged in burned mice infected with cytomegalovirus; Erickson EJ et al.; Cytomegalovirus is seen frequently in patients with burns and may enhance morbidity and mortality rates . Burned BALB/c mice that were given murine cytomegalovirus by intraperitoneal injection demonstrate increased bacterial translocation, as evidenced by positive mesenteric lymph node culture at 5 days after burn injury . The present experiment was conducted to investigate the time course and mechanism of this effect . A significant delay in the resolution of positive mesenteric lymph node cultures was observed with thermal injury alone . This was further reduced by the addition of murine cytomegalovirus infection to thermal injury . No such delay was seen with murine cytomegalovirus infection alone or in controls. Clin Chim Acta, 1991 Aug 30, 200(2-3), 211 - 9 Increased elastase-alpha 1-antitrypsin complex in fulminant hepatic failure: relationship to bacterial infection and activation of coagulation; Langley PG et al.; To study the effect of infection, a frequent complication of fulminant hepatic failure (FHF), on the release of elastase from polymorphonuclear leucocytes and its inhibition in circulation we have measured the concentrations of alpha 1-antitrypsin, which binds and inhibits elastase in the circulation, and of elastase-alpha 1-antitrypsin complex, in 30 patients with FHF . Elastase-alpha 1-antitrypsin complex was significantly increased in FHF as compared to controls (303 +/- 51 micrograms/l compared to 37 +/- 5 micrograms/l; n = 10; P less than 0.001) demonstrating activation of leucocytes in FHF . Infection caused greater release of leucocyte elastase, complex levels were significantly greater in patients who were infected when compared to those who were not (463 +/- 84 micrograms/l; n = 13 compared to 180 +/- 46 micrograms/l; n = 17; P less than 0.01) . Also patients who survived had significantly lower complex levels than those who did not (212 +/- 49 micrograms/l; n = 18 compared to 440 +/- 94 micrograms/l; n = 12; P less than 0.02) . alpha 1-Antitrypsin activity was not significantly different from control subjects (0.99 +/- 0.06 U/ml compared to 0.97 +/- 0.05 U/ml) . However alpha 1-antitrypsin activity was significantly higher in patients who survived (1.17 +/- 0.05 U/ml; n = 18) compared to those who did not (0.71 +/- 0.03 U/ml; n = 12; P less than 0.001) and patients who died had significantly lower levels than control subjects (P less than 0.01) indicating the importance of maintenance of normal inhibitor levels in patients with FHF . The leucocyte activation and release of elastase in FHF was linked to activation of the coagulation system; elastase--alpha 1-antitrypsin complex levels correlated significantly with thrombin-antithrombin III complex levels (r = 0.68; P less than 0.001) and inversely with fibrinogen (r = -0.71; P less than 0.001). Biochemistry, 1991 Aug 27, 30(34), 8315 - 22 Investigation into the source of electron transfer asymmetry in bacterial reaction centers; McDowell LM et al.; We have investigated the primary photochemistry of two symmetry-related mutants of Rhodobacter sphaeroides in which the histidine residues associated with the central Mg2+ ions of the two bacteriochlorophylls of the dimeric primary electron donor (His-L173 and His-M202) have been changed to leucine, affording bacteriochlorophyll (BChl)/bacteriopheophytin (BPh) heterodimers . Reaction centers (RCs) from the two mutants, (L)H173L and (M)H202L, have remarkably similar spectral and kinetic properties, although they are quite different from those of wild-type RCs . In both mutants, as in wild-type RCs, electron transfer to BPhL and not to BPhM is observed . These results suggest that asymmetry in the charge distribution of the excited BChl dimer (P*) in wild-type RCs (due to differing contributions of the two opposing intradimer charge-transfer states) contributes only modestly to the directionality of electron transfer . The results also suggest that differential orbital overlap of the two BChls of P with the chromophores on the L and M polypeptides does not contribute substantially to preferential electron transfer to BPhL. J Biol Chem, 1991 Aug 25, 266(24), 16171 - 7 In vitro regulation of human phagocyte cytochrome b heavy and light chain gene expression by bacterial lipopolysaccharide and recombinant human cytokines; Newburger PE et al.; We examined the effects of bacterial lipopolysaccharide and several recombinant human cytokines (tumor necrosis factor alpha and granulocyte-, macrophage-, and granulocyte-macrophage colony-stimulating factors) on the expression of the genes for the phagocyte cytochrome b, an essential component of the superoxide-generating oxidase . In vitro treatment with lipopolysaccharide, tumor necrosis factor alpha, or macrophage- or granulocyte-macrophage colony-stimulating factors increased the levels of transcripts for the cytochrome b heavy chain (gp91phox) 9- to 22-fold and transcripts for the light chain (p22phox) 2- to 5-fold in cultured human monocyte-derived macrophages . The same agents, except for macrophage colony-stimulating factor, induced the expression of the cytochrome b heavy chain gene 2- to 12-fold and light chain gene 2- to 6-fold in human granulocytes . The expression of the cytochrome b heavy and light chain genes was coordinated in both macrophages and neutrophils with regard to stimulus specificity and dose-response pattern . The time course for induction of the two genes was parallel in both cell types for all stimuli . The macrophage response to lipopolysaccharide occurred at least in part at the transcriptional level . These results show that a variety of physiological regulators modulate the coordinated expression of the cytochrome b genes. J Biol Chem, 1991 Aug 25, 266(24), 16044 - 9 Characterization and bacterial expression of the Dictyostelium myosin light chain kinase cDNA . Identification of an autoinhibitory domain; Tan JL et al.; A full-length cDNA corresponding to the Dictyostelium myosin light chain kinase gene has been isolated and characterized . Sequence analysis of the cDNA confirms conserved protein kinase subdomains and reveals that the Dictyostelium sequence is highly homologous to those of calcium/calmodulin-dependent protein kinases, including myosin light chain kinases from higher eukaryotes . Despite the high homologies to calcium/calmodulin-dependent protein kinases, there is no recognizable calmodulin-binding domain within the Dictyostelium sequence . However, the Dictyostelium myosin light chain kinase possesses a putative auto-inhibitory domain near its carboxyl terminus . To further characterize this domain, the full-length enzyme as well as a truncated form lacking this domain were expressed in bacterial cells and purified . The full-length enzyme expressed in bacteria exhibits essentially the same biochemical characteristics as the enzyme isolated from Dictyostelium . The truncated form however exhibits a Vmax that is approximately ten times greater than that of the native enzyme . In addition, unlike the native kinase and the full-length kinase expressed in bacteria, the truncated enzyme does not undergo autophosphorylation . These results suggest that the Dictyostelium enzyme, like myosin light chain kinases from higher eukaryotes, is regulated by an autoinhibitory domain but that the specific molecular signals necessary for activation of the Dictyostelium enzyme are entirely distinct. FEBS Lett, 1991 Aug 19, 288(1-2), 109 - 13 Probing the secondary quinone (QB) environment in photosynthetic bacterial reaction centers by light-induced FTIR difference spectroscopy; Breton J et al.; The photoreduction of the secondary electron acceptor, QB, has been characterized by light-induced Fourier transform infrared difference spectroscopy of Rb . sphaeroides and Rp . viridis reaction centers . The reaction centers were supplemented with ubiquinone (UQ10 or UQ0) . The QB- state was generated either by continuous illumination at very low intensity or by single flash in the presence of redox compounds which rapidly reduce the photooxidized primary electron donor P+ . This approach yields spectra free from P and P+ contributions making possible the study of the microenvironment of QB and QB- . Assignments are proposed for the C...O vibration of QB- and tentatively for the C = O and C = C vibrations of QB. Biochem Biophys Res Commun, 1991 Aug 15, 178(3), 1188 - 93 Individual alpha and beta subunits of bacterial luciferase exhibit bioluminescence activity; Waddle J et al.; The alpha subunit of luciferase, encoded by the luxA gene, and the beta subunit, encoded by the luxB gene, have been expressed independently in Escherichia coli . We have found that the individual subunits of the heterodimeric (alpha beta) enzyme have bioluminescence activity . Overexpression of the individual subunits in Escherichia coli at 37 degrees resulted in accumulation of large amounts of the specific subunit in the insoluble fraction, while expression at lower temperatures (20 degrees-26 degrees) resulted in accumulation of high levels of the individual subunits in the soluble fraction of the cell lysates . We were surprised to find that addition of n-decanal to extracts of cells carrying either the luxA gene or the luxB gene followed by injection of FMNH2, conditions of the standard luciferase assay, resulted in emission of significant bioluminescence . The occurrence of the activity in lysates of E . coli cells carrying the gene for only one subunit showed that the activity of the isolated subunits is not due to contamination of one subunit with trace amounts of the other . The observed bioluminescence activity is the result of reactions catalyzed by the alpha subunit in the absence of beta subunit and also by the beta subunit in the absence of the alpha subunit. Proc Natl Acad Sci U S A, 1991 Aug 15, 88(16), 7204 - 8 Systemic expression of a bacterial gene by a tobacco mosaic virus-based vector; Donson J et al.; Tobacco mosaic virus (TMV) produces large quantities of RNA and protein on infection of plant cells . This and other features, attributable to its autonomous replication, make TMV an attractive candidate for expression of foreign sequences in plants . However, previous attempts to construct expression vectors based on plant RNA viruses, such as TMV, have been unsuccessful in obtaining systemic and stable movement of foreign genes to uninoculated leaves in whole plants . A hybrid viral RNA (TB2) was constructed, containing sequences from two tobamoviruses (TMV-U1 and odontoglossum ringspot virus) . Two bacterial sequences inserted independently into TB2 moved systemically in Nicotiana benthamiana, although they differed in their stability on serial passage . Systemic expression of the bacterial protein neomycin phosphotransferase was demonstrated . Hybrid RNAs containing both TMV-U1 and the inserted bacterial gene sequences were encapsidated by the odontoglossum ringspot virus coat protein, facilitating their transmission and amplification on passaging to subsequent plants . The vector TB2 provides a rapid means of expressing genes and gene variants in plants. J Mol Evol, 1991 Aug, 33(2), 179 - 93 Evolutionary relationships among the permease proteins of the bacterial phosphoenolpyruvate: sugar phosphotransferase system . Construction of phylogenetic trees and possible relatedness to proteins of eukaryotic mitochondria; Reizer A et al.; The amino acid sequences of 15 sugar permeases of the bacterial phosphoenolpyruvate-dependent phosphotransferase system (PTS) were divided into four homologous segments, and these segments were analyzed to give phylogenetic trees . The permease segments fell into four clusters: the lactose-cellobiose cluster, the fructose-mannitol cluster, the glucose-N-acetylglucosamine cluster, and the sucrose-beta-glucoside cluster . Sequences of the glucitol and mannose permeases (clusters 5 and 6, respectively) were too dissimilar to establish homology with the other permeases, but short regions of statistically significant sequence similarities were noted . The functional and structural relationships of these permease segments are discussed . Some of the homologous PTS permeases were found to exhibit sufficient sequence similarity to subunits 4 and 5 of the eukaryotic mitochondrial NADH dehydrogenase complex to suggest homology . Moreover, subunits 4 and 5 of this complex appeared to be homologous to each other, suggesting that these PTS and mitochondrial proteins comprise a superfamily . The integral membrane subunits of the evolutionarily divergent mannose PTS permease, the P and M subunits, exhibited limited sequence similarity to subunit 6 of the mitochondrial F1F0-ATPase and subunit 5b of cytochrome oxidase, respectively . These results suggest that PTS sugar permeases and mitochondrial proton-translocating proteins may be related, although the possibility of convergent evolution cannot be ruled out. J Am Acad Dermatol, 1991 Aug, 25(2 Pt 1), 287 - 99 Sexually transmitted diseases: bacterial infections . Committee on Sexually Transmitted Diseases of the American Academy of Dermatology; Buntin DM et al.; This update reviews the latest diagnostic and therapeutic methods regarding sexually transmitted diseases caused by bacteria . The following mucocutaneous bacterial disorders will be discussed: syphilis, chancroid, lymphogranuloma venereum, granuloma inguinale, and gonorrhea. Pediatr Ann, 1991 Aug, 20(8), 413 - 8 Bacterial respiratory infections and otitis media; Shapiro ED; Bacterial infections of the respiratory tract are an important potential problem for children who attend group day care . Immunization (for Hib) or chemoprophylaxis, when appropriate, should lessen the risk and help to control or prevent the problem in many instances. J Pediatr Surg, 1991 Aug, 26(8), 921 - 4 Intralipid adversely affects reticuloendothelial bacterial clearance; Katz S et al.; Lipid emulsion is a major caloric source in patients receiving total parenteral nutrition . Cleared by the Kupffer cells of the reticuloendothelial system (RES), lipid emulsion may adversely affect the RES function by decreasing its ability to remove blood-borne bacteria . This study evaluates and compares the blood clearance and organ localization of viable radiolabeled {35S} Escherichia coli following slow intraperitoneal (IP) and more rapid intravenous (IV) administration of a 20% fat emulsion (FE) . Sixty male Sprague-Dawley rats weighing 150 g were placed in 6 experimental groups (10 rats per group) . Group 1 received IP normal saline (3 mL/d for 3 days); group II received IP FE (20%) (4 g/kg/d for 3 days); and group III received normal saline IV (3 mL/d for 3 days) . The remaining animals received a slow (15-minute) IV infusion of FE (4 g/kg/dose) prior to bacterial challenge: group IV at 4 hours; group V at 24 hours; and group IV at 4 and 24 hours . E coli (10(9)/mL) were injected via the tail vein . Blood samples were obtained for clearance study . At 10 minutes, tissue samples (50 to 100 mg) of liver, spleen, kidney, and lung were obtained and processed for liquid scintillation counting . Although rapid bacterial blood clearance was found in all the groups, there was a significant change in organ localization of bacteria . Normal distribution of bacteria in group I was as follows: liver 70.1% +/- 6.2%, spleen 5.2% +/- 1.2%, kidney 0.2% +/- 0.04%, and lung 1.6% +/- 0.6% . There was a slight increase in lung localization of bacteria in rats receiving IP FE (3.7% +/- 1.5%; P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS) Nippon Yakurigaku Zasshi, 1991 Aug, 98(2), 63 - 71 {The contractile response of isolated lingual arteries from rabbits treated with bacterial lipopolysaccharide via the stimulation of B1-receptors for kinins}; Sugihara M et al.; The effect of intravenous injection of 10 micrograms of a lipopolysaccharide (endotoxin) extracted from E . coli to rabbits on the responses of isolated lingual arteries to des-Arg9-bradykinin (a specific kinins B1-receptor agonist) was studied . Endotoxin injection led to the appearance of the endothelium-independent contractile effect of des-Arg9-bradykinin in the arteries; endotoxin elicited this response when administered at 1, 5 or 20 hr before the experiment, in a time-interval dependent manner . The contractile response to des-Arg9-bradykinin of the arteries isolated from animals receiving endotoxin 20 hr before the experiment was attenuated by des-Arg9-{Leu8}-bradykinin (a specific inhibitor of kinins B1-receptor) or pretreatment of the animals with an inhibitor of protein synthesis (cycloheximide and actinomycin D) . When compared with the effect of des-Arg9-bradykinin, bradykinin (a potent kinin B2-receptor, but weak B1-receptor stimulant) caused slight contraction of the arteries; however, this effect was not endotoxin-dependent and was not modified by des-Arg9-{Leu8}-bradykinin . Effect of in vitro preincubation with endotoxin of the arteries isolated from animals receiving saline 20 hr before the experiment was further studied . The preincubation (for 1 and 5 hr) with endotoxin of the arteries in the presence or absence of plasma had no effect on the sensitivity of the arteries to des-Arg9-bradykinin; the sensitivity was also unaffected in the presence or absence of endotoxin, thus suggesting that there is no interaction between endotoxin and some plasma-related factors with the appearance of the contraction in response to the kinin B1-receptor agonist in the arteries in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) J Immunol, 1991 Aug 1, 147(3), 980 - 3 Activation by bacterial lipopolysaccharide causes changes in the cytosolic free calcium concentration in single peritoneal macrophages; Letari O et al.; Variations in the cytosolic free Ca2+ concentration {( Ca2+}i) upon LPS exposure were studied in single rat peritoneal macrophages loaded with fura-2 under carefully controlled conditions . Of a total of 60 cells examined, 47% responded to LPS (1 microgram/ml) with an increase in {Ca2+}i . Macrophages were heterogeneous with regard to the LPS response, with individual cells exhibiting single rapid and transient increases in {Ca2+}i, multiple transients, or slower and more sustained variations . In 62% of the responding cells, a second exposure to LPS elicited a {Ca2+}i rise, although usually to a slightly lower peak value . Thus, rapid desensitization to LPS does not occur in the majority of these macrophages . EGTA did not abolish the response of those cells that exhibited a single rapid transient in {Ca2+}i, indicating that the source of the initial {Ca2+}i rise was the intracellular stores . There was no obvious correlation between the type of response to LPS and the initial morphologic features (rounded vs polarized) of the cells . Our present work shows unequivocally that LPS induces increases in macrophage {Ca2+}i and, thereby, lends substantial support to the hypothesis that {Ca2+}i is a second messenger in LPS-mediated activation of the macrophage. J Cell Biol, 1991 Aug, 114(4), 609 - 21 Characterization of YDJ1: a yeast homologue of the bacterial dnaJ protein; Caplan AJ et al.; The YDJ1 (yeast dnaJ) gene was isolated from a yeast expression library using antisera made against a yeast nuclear sub-fraction termed the matrix lamina pore complex . The predicted open reading frame displays a 32% identity with the sequence of the Escherichia coli heat shock protein dnaJ . Localization of YDJ1 protein (YDJ1p) by indirect immunofluorescence reveals it to be concentrated in a perinuclear ring as well as in the cytoplasm . YDJ1p cofractionates with nuclei and also microsomes, suggesting that its perinuclear localization reflects association with the ER . YDJ1p is required for normal growth and disruption of its gene results in very slow growing cells that have pleiotropic morphological defects . Haploid cells carrying the disrupted YDJ1 gene are inviable for growth in liquid media . We further show that a related yeast protein, SIS1, is a multicopy suppressor of YDJ1. Am J Pathol, 1991 Aug, 139(2), 371 - 82 Mesangial cell activation by bacterial endotoxin . Induction of rapid cytoskeletal reorganization and gene expression; Bursten SL et al.; Cultured glomerular mesangial cells (MC) respond to low concentrations of bacterial endotoxin (ET) by secreting prostaglandins and interleukin-1 . To evaluate further the nature of ET-induced mesangial cell activation, the authors evaluated the effects of this agent on MC morphology and cytoskeletal organization . Bacterial ET, in concentrations as low as I ng/ml, induced reversible membrane ruffling, cellular rounding, and extension of many filopodia and lamellopodia . Augmented fluid-phase pinocytosis occurred in parallel, as determined by transmission electron microscopy and tritiated sucrose uptake . These cellular morphologic and functional changes were associated with an extensive, but reversible, depolymerization of actin microfilaments . Actin gene expression was also modified by ET . At 4 to 6 hours after ET exposure, Northern blot analysis showed a twofold to fourfold increase in actin mRNA levels . In situ hybridizations of ET-stimulated cells at the light and electron microscopic levels demonstrated a markedly asymmetric distribution of actin mRNA, which was localized in the cellular periphery at filopodial and lamellopodial extensions, presumably sites of new actin protein synthesis . It is concluded that ET effects on MC are distinct from the nonspecific lytic or 'toxic' actions described for other cell types . Endotoxin induces a global activation of this cell type associated with major changes in membrane structure, cytoskeletal organization, and gene expression, which resemble in many respects the responses to peptide mitogens. Clin Exp Immunol, 1991 Aug, 85(2), 270 - 7 Bacterial lipopolysaccharide enhances deposition of immune complexes and exacerbates nephritis in BXSB lupus-prone mice; Granholm NA et al.; Systemic autoimmune disease is influenced by genetic, immunological, hormonal, and environmental factors . Although environmental factors are major agents that induce or exacerbate autoimmune diseases, the mechanism(s) and the molecular events by which they operate remain poorly understood . Here we used the lupus-prone BXSB mouse as an animal model of systemic autoimmune disease, and we used a bacterial lipopolysaccharide (LPS) as a surrogate infectious agent to gain some insight into the mechanism(s) by which infectious agents exacerbate autoimmune diseases . Our experimental protocol was designed to address three questions: (i) whether spontaneous polyclonal B cell activation (PBA) that occurs in BXSB mice could be further enhanced by bacterial LPS; (ii) whether repeated exposure to LPS would exacerbate autoimmune disease, as reflected by enhanced deposits of immune complexes (ICs) in kidneys and exacerbated nephritis; and (iii) whether the mechanism by which LPS exacerbates nephritis might involve interference with blood cell carrier function, mononuclear phagocyte function, or both . BXSB mice were injected with LPS (25 micrograms) twice a week for 5 weeks; control autoimmune BXSB mice and immunologically normal (C57BL/6) mice were injected with vehicle only . The three groups of mice were then challenged with soluble ICs to assess the kinetics of their disappearance from the circulation, their uptake by the mononuclear phagocyte system (liver, spleen), their distribution in target organ (kidney), and blood cell carrier function . The results indicate that: (i) spontaneous PBA can be enhanced further by LPS; (ii) exposure to LPS results in increased deposits of endogenous ICs in kidneys and exacerbated nephritis; and (iii) defective handling of ICs by the mononuclear phagocyte system and impaired blood cell carrier function are contributory factors to exacerbated nephritis, but that mechanisms in addition to passive localization of ICs may also be operative. Hepatology, 1991 Aug, 14(2), 313 - 7 Bacterial invasion of the biliary system by way of the portal-venous system; Sung JY et al.; It has been suggested that bacteria in the intestine gain access into the biliary tract by entering the portal-venous blood . We have tested the hypothesis of hematogenous infection of the biliary system in cats . The animals were treated in three different groups: group A (no biliary obstruction), group B (acute biliary obstruction) and group C (chronic biliary obstruction) . A mutant strain of Escherichia coli was infused into the splenic vein of cats at three different dosages (10(7), 10(5) and 10(3) with sham controls . In the unobstructed biliary system, the mutant E . coli was isolated from the bile 30 min and 90 min after the infusion of 10(7) and 10(5) E . coli, respectively . No bacteria were found in the bile with the infusion of 10(3) E . coli and in the control animals . Bile flow was significantly reduced with the infusion of bacteria . The biliary excretion of E . coli in group B was similar to that in group A . In group C, the bile output in the first hour was very high but declined rapidly . E . coli was excreted into the bile at all three dosages of infusion after 30 min . Histological sections of the liver showed that the infused bacteria entered the sinusoidal blood and that some were phagocytosed by Kupffer cells . The portal-venous blood was considered an important route of bacterial invasion into the biliary system, and the penetration of bacteria was facilitated in biliary obstruction. AJR Am J Roentgenol, 1991 Aug, 157(2), 365 - 70 Bacterial osteomyelitis: findings on plain radiography, CT, MR, and scintigraphy; Gold RH et al.; Early detection of osteomyelitis is essential if appropriate therapy is to be started before bone devitalization . Although the 99mTc-methylene diphosphonate (MDP) bone scan may signify the possibility of osteomyelitis days or weeks before osseous changes are apparent on standard radiographs, the radiographic changes may provide important diagnostic clues . The 67Ga-citrate scan augments the diagnostic value of the 99mTc-MDP scan, and the 111In-labeled WBC scan is useful for detecting abscesses . CT aids in the detection of sequestra, and MR imaging is useful in defining the extent of the inflammatory process and in distinguishing osteomyelitis from cellulitis . In this article, we review and correlate the changes of bacterial osteomyelitis shown by these imaging methods. Int J Androl, 1991 Aug, 14(4), 264 - 70 Is the sperm bacterial ratio a determining factor in impairment of sperm motility: an in-vitro study in man with Escherichia coli; Auroux MR et al.; The effects of urogenital infection on male fertility are controversial . The object of this study was to assess whether contact between E . coli, one of the bacteria encountered most frequently in semen cultures, and sperm was involved in decreasing motility of the sperm . Sperm from healthy donors were therefore incubated at two concentrations (1.10(7) and 4.10(7) ml-1) with bacteria (10(4) and 10(6) bacteria ml-1 respectively) . Sperm motility was assessed as a function of time . The endotoxin effect was also evaluated . Aliquots of the sperm were used as controls . The motility of a population of 10(6) sperm ml-1 was reduced significantly more by the presence of 10(6) ml-1 E . coli than a sperm population four times more numerous . Since the endotoxin had no effect on sperm motility, it is possible this phenomenon is due to bacterial adherence to the sperm . From this study, it is therefore probable that the presence of E . coli in semen decreases sperm motility, but that this depends on the sperm:bacterial ratio ml semen-1. Prenat Diagn, 1991 Aug, 11(8), 551 - 7 Risk of miscarriage after transcervical and transabdominal CVS in relation to bacterial colonization of the cervix; Baumann P et al.; Cervico-genital colonization with micro-organisms poses a potential threat to the pregnancy when transcervical (TC) CVS is performed . In order to evaluate this threat, cervical swabs in 478 patients were obtained and cultured for bacteria, yeasts, and mycoplasmas; chlamydias were detected by an enzyme immunology test . Two hundred and seventy-one patients had CVS done transvaginally and 207 underwent transabdominal (TA) CVS . Transvaginal specimens were obtained in 61.6 per cent by forceps biopsy . Overall in 29.9 per cent of patients micro-organisms were detected, the rate and distribution of different species being the same in both groups . There were 36 (7.5 per cent) miscarriages up to 28 weeks of gestation in the combined groups, 29 (10.7 per cent) in the TC-CVS group and 7 (3.4 per cent) in the TA-CVS group . When miscarriages occurred after TC-CVS, bacteria/yeasts were involved in 10.3 per cent of cases and mycoplasmas in 37.9 per cent, this proportion being almost the same in early (less than 2 weeks) and late (greater than 2 weeks) miscarriages . After TA-CVS, in 28.6 per cent only mycoplasmas, and this only in late miscarriages (greater than 2 weeks), were involved, accounting for 40 per cent of late miscarriages.
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