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