Neuroimmunomodulation, 1994 Jul-Aug, 1(4), 251 - 8 An interleukin-1-alpha-like neuronal system in the preoptic-hypothalamic region and its induction by bacterial lipopolysaccharide in concentrations which alter pituitary hormone release; Rettori V et al.; We studied the effect of intravenous injection of lipopolysaccharide (LPS) (30-250 micrograms) on the release of several anterior pituitary hormones as indicated by changes in their concentrations in plasma . Within 30 min after intravenous injection of LPS there was a dose-related stimulation of ACTH release; prolactin (PRL) release was induced only by the highest LPS dose injected (250 micrograms) . Even the lowest dose of LPS (30 micrograms) decreased plasma growth hormone (GH) by 60 min . Higher doses lowered plasma GH by 30 min, but thyroid-stimulating hormone release was only significantly inhibited by the highest dose of LPS . The action of LPS seems to be primarily exerted on the central nervous system, since incubation of hemipituitaries with LPS for 3 h in doses ranging from 0.001 to 10 micrograms/ml had no effect on ACTH release . LPS is thought to induce its effects on hormones either by release of cytokines from immune cells which subsequently induce the hormonal changes or possibly by direct action within the hypothalamus . In this report we demonstrate the immunocytochemical localization of a population of interleukin-1 alpha (IL-1 alpha)-like cells in a region extending from the basal forebrain at the level of the diagonal band of Broca, caudally and dorsally to the dorsolateral preoptic region and the hypothalamus at the level of the paraventricular nucleus . Further caudally, IL-1 alpha-like immunoreactive cells were located in the midportion of the amygdala . Two hours after injection of the 125-micrograms dose of LPS, the number of these immunoreactive cells was dramatically increased.(ABSTRACT TRUNCATED AT 250 WORDS) Biochim Biophys Acta, 1994 Jun 30, 1222(2), 171 - 8 Monitoring of bacterial growth and structural analysis as probed by FT-IR spectroscopy; Zeroual W et al.; Fourier-transform infrared spectroscopy was used to explore structural changes in bacteria under different incubation conditions . In particular, differences between Bradyrhizobium japonicum (BRJ) grown in liquid and on solid media were investigated, as well as the rearrangement of BRJ after transfer from one medium to the other . The FT-IR absorption bands located between 1200 and 900 cm-1 region, vary in spectral shape and intensity when BRJ were suspended in solution medium or plated on solid medium . In agreement with the electronic micrograph data, these spectroscopic changes are due to the changes involving the bacterial wall (peptidoglycan) when BRJ are plated in agar medium . By means of this FT-IR ultrastructural study of Bradyrhizobium japonicum bacteria, it has been possible to follow and to evaluate the rate of the molecular change in bacteria without any destructive interference . This indicates that FT-IR spectroscopy can prove to be a valuable technique in the monitoring of metabolic events in bacterial cells relevant to agriculture as well as environmental and health sciences. Cancer Lett, 1994 Jun 15, 81(1), 99 - 109 In vitro TNF-alpha production and in vivo alteration of TNF-alpha RNA in mouse peritoneal macrophages after treatment with different bacterial derived agents; Novakovic S et al.; Since muramyl dipeptide (MDP) was recognized as a potent monocyte/macrophage activating agent, many MDP analogues were synthesized and tested for their ability to augment the host immune defence system against neoplasms . This study was performed to determine whether the newly synthesized desmuramyl N-acyl dipeptides LK 409 and LK 410 were also capable of affecting the immune system . For this purpose, the peritoneal macrophages were incubated in vitro with these two agents and TNF-alpha production was measured . In addition, the effect of LK 409 and LK 410 on TNF-alpha and IL-1 RNA levels in in vivo stimulated macrophages was determined by quantitative polymerase chain reaction (RT-PCR) . None of the LK 409 and LK 410 concentrations tested were able to render macrophages in vitro to excrete a detectable amount of TNF-alpha in the supernatant fluid . However, the TNF-alpha and IL-1 RNA levels in macrophages of in vivo treated mice (C57Bl/6) were increased in comparison to mock-treated mice . The results indicate that LK 409 and LK 410 are capable of inducing an increase in TNF-alpha and IL-1 RNA levels, yet in vitro TNF-alpha production remains under detectable levels (40 U/ml). J Immunol, 1994 Jun 15, 152(12), 5660 - 9 Molecular context of a viral T cell determinant within a chimeric bacterial protein alters the diversity of its T cell recognition; Lo-Man R et al.; We genetically introduced two different viral CD4+ T cell epitopes within two internal sites of the Escherichia coli maltose-binding (MalE) protein . Affinity-purified hybrid MalE proteins were used to analyze the influence of the molecular environment on the presentation of inserted epitope to T cells . In the first model, the 120 to 132 PreS T cell epitope was inserted alone or with its C-terminal B cell epitope (132-145) at site 133 or 303 of MalE . The maltose-binding protein with PreS peptide inserts expressing the 120 to 132 sequence were able to induce in vivo and in vitro peptide-specific T cell response, whatever the length and the position of the insert . In the second model, the 103 to 115 T cell epitope from the C3 region of poliovirus type 1 (PV1) was inserted, with various flanking sequences, either at site 133 or 303 of MalE protein . The longer C3:86 to 115 insert induced poliovirus-specific T cell responses at both sites of MalE, whereas the C3:93 to 115 insert did it only at site 303 but not at site 133 . Moreover, C3:103 to 115 specific T cell hybridomas discriminated between the processed peptides generated from the different chimeric proteins, as a result of differences in the length and the position of the inserted sequence . Therefore, in this experimental model the loss of in vivo immunogenicity of an antigenic determinant within a chimeric protein is related to the activation of a reduced T cell repertoire . These observations involve important consequences for the engineering of recombinant vaccines. Gene, 1994 Jun 10, 143(2), 171 - 7 RNA- and single-stranded DNA-binding (SSB) proteins expressed during Drosophila melanogaster oogenesis: a homolog of bacterial and eukaryotic mitochondrial SSBs; Stroumbakis ND et al.; Little is known about the identity and involvement of single-stranded (ss) DNA-binding (SSB) and RNA-binding proteins in developmental processes that occur during oogenesis in Drosophila melanogaster (Dm) . Here, we describe a molecular approach designed to identify such proteins by virtue of their ssDNA-binding activity . We have constructed a directional ovarian cDNA library and conducted expression cloning screens which identified five unique cDNAs that encode proteins capable of binding ssDNA . All five represent previously unreported sequences . The remainder of this paper focuses on one of these cDNAs which encodes a Dm protein displaying significant sequence homology to Escherichia coli ssDNA-binding protein (SSB, involved in DNA replication, repair and recombination), as well as eukaryotic SSBs isolated from the mitochondria (mt) of rats, frogs, humans and yeast . The deduced amino acid (aa) sequence of this 15.6-kDa protein, which we will refer to as Dm mtSSB, displays average identities of 38.3% with eukaryotic mtSSBs and 23.4% with bacterial SSBs . Gel retardation analysis with an affinity-purified GST fusion protein confirms that Dm mtSSB specifically binds ss, but not double stranded DNA . Dm mtSSB is encoded by a nuclear gene whose expression appears to be developmentally regulated . It is expressed as a single 600-nucleotide (nt) transcript during oogenesis and embryogenesis . A larger transcript of 1500 nt is prevalent in some later stages of Dm development. J Bacteriol, 1994 Jun, 176(11), 3140 - 7 Genetic analysis of a plasmid-encoded, host genotype-specific enhancement of bacterial fitness; Lenski RE et al.; In the absence of antibiotics, carriage of pACYC184 reduces the competitive fitness of an Escherichia coli B genotype that was not previously selected for plasmid carriage, relative to that of an isogenic plasmid-free competitor . However, a host genotype propagated with the plasmid for 500 generations evolved an unexpected competitive advantage from plasmid carriage, relative to its own isogenic plasmid-free segregant . We manipulated the pACYC184 genome in order to identify the plasmid-encoded function that was required for the enhancement of the coevolved host genotype's competitive fitness . Inactivation of the plasmid-encoded tetracycline resistance gene, by deletion of either the promoter region or the entire gene, eliminated the beneficial effect of plasmid carriage for the coevolved host . This beneficial effect for the coevolved host was also manifest with pBR322, which contains a tetracycline resistance gene identical to that of pACYC184 but is otherwise heterologous. Infect Immun, 1994 Jun, 62(6), 2195 - 201 Differential priming effects of proinflammatory cytokines on human neutrophil oxidative burst in response to bacterial N-formyl peptides; Elbim C et al.; Cytokines such as tumor necrosis factor alpha (TNF-alpha), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin 8 (IL-8), IL-6, IL-1 alpha, and IL-1 beta produced during the immune and inflammatory responses to bacterial stimuli have been reported to interact with polymorphonuclear neutrophil (PMN) activities . However, contradictory findings on their direct and priming effects on the PMN oxidative burst, which is essential for bacterial killing, have been reported . We have used a flow cytometry method to study the effects of these cytokines on the oxidative burst of PMN in whole blood to avoid PMN activation related to isolation procedures . None of the cytokines tested directly activated the PMN oxidative burst, but they did have differential priming effects on the oxidative burst in response to bacterial N-formyl peptides . TNF, GM-CSF, and IL-8 strongly primed a subpopulation of PMN to produce H2O2 in response to N-formyl-methionyl-leucyl-phenylalanine (FMLP), while IL-1 alpha, IL-1 beta, and IL-6 failed to do so . Furthermore, the addition of TNF, GM-CSF, or IL-8 to whole blood increased the capacity of a subpopulation of PMN to bind N-formyl peptides, a phenomenon that could account, at least in part, for the strong H2O2 production in response to FMLP after priming by the cytokines . The size of the primed hyperresponsive subpopulation was greater after priming with TNF or GM-CSF than after priming with IL-8 . However, GM-CSF, TNF, and IL-8 at suboptimal concentrations cooperated in the induction of a subpopulation hyperresponsive to FMLP . These results show that, of the various proinflammatory cytokines tested, TNF, GM-CSF, and IL-8 strongly prime the PMN oxidative burst in response to bacterial peptides in whole blood and suggest that these cytokines may play a critical role in bacterial killing in vivo. Pediatr Infect Dis J, 1994 Jun, 13(6), 477 - 84 Crossover of placebo patients to intravenous immunoglobulin confirms efficacy for prophylaxis of bacterial infections and reduction of hospitalizations in human immunodeficiency virus-infected children . The National Institute of Child Health and Human Development Intravenous Immunoglobulin Clinical Trial Study Group; Mofenson LM et al.; After completion of a placebo-controlled trial of intravenous immunoglobulin (IVIG) infection prophylaxis, patients were offered open label IVIG and optional participation in a follow-up study . The purpose of the follow-up study was to evaluate the IVIG effect in original placebo recipients and longevity of IVIG benefit in original IVIG recipients . Of 212 human immunodeficiency virus-infected children on study at trial closure, 148 (67 of 98 (68%) placebo and 81 of 114 (71%) IVIG patients) received open label IVIG for a mean of 16 months . When open label IVIG was begun, 45% were receiving trimethoprim-sulfamethoxazole prophylaxis for Pneumocystis carinii pneumonia (43% of placebo and 47% of IVIG patients) and 54% were receiving zidovudine (55% of placebo and 53% of IVIG patients) . In patients who received placebo during the original study, the rate of serious bacterial infections was significantly lower after change to open label IVIG (estimated 15.8 fewer episodes/100 patient years; 95% confidence interval, 3.2 to 28.5; P = 0.014) . Similar findings were observed for minor bacterial infections (estimated 61.2 fewer/100 patient years; 95% confidence interval, 29.2 to 93.3; P < 0.001) and hospitalizations (estimated 43.7 fewer/100 patient years; 95% confidence interval, 27.7 to 59.6; P < 0.001) . Decreases were observed whether or not trimethoprim-sulfamethoxazole prophylaxis was being given at the time open label IVIG was begun . In patients who received IVIG during the original study, no significant difference was seen in infections or hospitalizations after change to open label IVIG.(ABSTRACT TRUNCATED AT 250 WORDS) Curr Opin Pediatr, 1994 Jun, 6(3), 327 - 33 Bacterial translocation in the neonate; Van Camp JM et al.; Bacterial translocation has been a major topic of investigation for the past two decades . Despite recent evidence that bacterial translocation may play a significant role in the morbidity and mortality of adults faced with multiple types of stress, very little is known about the effect of bacterial translocation on the neonate . Recently more and more evidence has suggested that normal as well as premature or ill neonates experience spontaneous bacterial translocation quite commonly . This article reviews the recent literature on bacterial translocation in stressed adults, the development of an intact intestinal mucosal barrier in the newborn as a protection against bacterial translocation, and the role of spontaneous bacterial translocation in the development of systemic sepsis and its accompanying morbidity and mortality. Plant J, 1994 Jun, 5(6), 873 - 84 Carbohydrate binding activities of Bradyrhizobium japonicum . III . Lectin expression, bacterial binding, and nodulation efficiency; Ho SC et al.; In previous studies, evidence that the Bradyrhizobium japonicum lectin, designated BJ38, mediated the observed carbohydrate-specific binding activities of the bacteria, including the saccharide-specific adhesion to soybean root cells was presented . In the present study, it is found that both B . japonicum, as well as the purified BJ38, bind predominantly to young emergent root hairs of soybean roots and, to a much lesser extent, to the root cap, mature root hairs, epicotyl or hypocotyl regions . Thus, the region of preferential binding for both the bacteria and the isolated lectin coincide with the region of the soybean root most susceptible to B . japonicum infection . The importance of bacterial binding for the nodulation process was studied by comparing the nodulation efficiency of binding-deficient mutants N4 and N6 to the wild-type . These mutants had been shown to be defective in carbohydrate recognition, as represented by their diminished ability to bind to soybean roots . BJ38 was immunolocalized to one pole of the cell surface of wild-type B . japonicum, but no surface labeling could be detected on either mutant . Moreover, both N4 and N6 showed a substantial decrease in nodulation activity, relative to the wild-type . These results provide additional evidence that the carbohydrate binding activity of B . japonicum, most probably mediated by BJ38, may play an important role(s) in the initial phases of the infection process. Burns, 1994 Jun, 20(3), 220 - 5 Supplementation of an elemental enteral diet with alanyl-glutamine decreases bacterial translocation in burned mice; Tenenhaus M et al.; Although there are many reports of the importance of early enteral feeding in maintaining gastrointestinal integrity and preventing bacterial translocation (BT) following burn injury, no diet has been shown clinically to protect the GI tract postburn . Several studies suggest that glutamine (GLN) may benefit gut integrity following injury, shock and other stress . Unfortunately, the free amino acid GLN is unstable in solution . Alanyl-glutamine (ALA-GLN), a soluble form of GLN, maintains long-term stability in solution and could be supplemented to conventional liquid enteral diets . We studied the effects of ALA-GLN supplementation of the elemental diet Vivonex TEN on effecting BT in mice following 32 per cent TBSA full skin thickness burns . Groups A-D were burned . Group A (30 mice) was fed standard rodent chow, which contains extremely high (clinically non-useable) levels of protein . Group B (51 mice) was fasted 24 h, then fed chow 24 h . Group C (64 mice) was fed Vivonex TEN, and Group D (65 mice) received Vivonex TEN plus ALA-GLN (GLN equivalent, 14 g/l) . A control group (Group E) consisted of 22 normal mice (no burn injury, chow diet) . Mice were assessed for BT by sterile harvesting and plating of mesenteric lymph node tissue, 48 h postburn . Plates were considered positive if any bacterial growth was noted . Non-burned mice exhibited no BT, while burn-fasted mice showed a 64.3 per cent incidence of BT (P = NS).(ABSTRACT TRUNCATED AT 250 WORDS) Med Sci Sports Exerc, 1994 Jun, 26(6), 687 - 94 Heat stress does not sensitize rats to the toxic effects of bacterial lipopolysaccharide; Ryan AJ et al.; To determine whether heat stress sensitizes rats to lipopolysaccharide (LPS), four groups were examined: saline, LPS, heat stressed+saline, and heat stressed+LPS treated rats . Saline or LPS (Escherichia coli, 5 mg.kg-1 body weight, i.v.) was given after exposure to heat and at the same time of day for nonheated rats . Survival was monitored for 24 or 48 h; samples of liver and small intestine were obtained at 24 h for histological analysis . Thermal responses were similar (P > 0.05) for the heat stressed saline and LPS treated rats: mean values for maximum colon temperature were 43.0 +/- 0.1 and 42.9 +/- 0.1 degrees C, respectively . Mortality was similar for rats exposed to heat stress+saline (11%, 2/19) and heat stress+LPS (32%, 6/19) . No lethality was observed in nonheated rats given saline or LPS . Tissue damage was similar in heat stress+saline and heat stress+LPS treated rats . Liver showed mild to severe degrees of coagulative necrosis while duodenum exhibited damage to the villous tips . These findings show that severe heat stress does not markedly sensitize the rat to the lethal activity of LPS. Can J Microbiol, 1994 Jun, 40(6), 426 - 31 Virulence of patient and water isolates of Legionella pneumophila in guinea pigs and mouse L929 cells varies with bacterial genotype; Bezanson G et al.; Thirteen isolates of Legionella pneumophila serogroup 1 (five from patients, eight from water) were screened for their virulence in guinea pigs after intraperitoneal injection and for their infectivity in L929 mouse cells . Since these isolates included three monoclonal antibody subtypes and four genotypes, the relative influence of these parameters on the pathogenicity of naturally occurring L . pneumophila could be assessed . There was no correlation between infectivity in the L929 assay and virulence for guinea pigs . The source of the isolate, patient or environmental, as well as the isolate's monoclonal antibody subtype did not correlate with virulence . At the p < 0.05 level, isolates with genotype IIb (20-MDa plasmid and EcoRI fragmentation pattern b) were significantly more virulent (mean log LD50 6.84) than genotype VIb (100-MDa plasmid, pattern b), IIId (72- and 96-MDa plasmids, pattern d) or Oc (no detectable plasmid, pattern c) isolates . Genotype IIId isolates were the least virulent (mean log LD50 9.49) . Plasmid-containing isolates were more infective than plasmidless ones in L929 cells (p = 0.0001) . We conclude that our strain types of L . pneumophila exhibit a gradation in virulence for guinea pigs and that infectivity in L929 cells does not correlate with virulence for guinea pigs. J Appl Bacteriol, 1994 Jun, 76(6), 608 - 15 A substrate-mediated assay of bacterial proton efflux/influx to predict the degree of spoilage of beef mince stored at chill temperatures; Seymour IJ et al.; A method was developed to predict spoilage of minced meat at chill temperatures, based on the difference in proton efflux from and influx into bacterial cells . This difference depends on the number of organisms present, the available glucose in the meat sample and the ability of the organisms to metabolize amino acids . The proton efflux/influx of a meat filtrate containing bacteria was measured at 25 degrees C with a pH/ion meter in the presence of peptone with or without glucose . There was a noticeable rate of change of mV h-1 of the meat filtrate prior to the organoleptic detection of spoilage which may be used semi-predictively to determine the remaining shelf-life of meat at different storage temperatures . The method could be investigated further, encompassing type and relative numbers of organisms, incubation temperature, meat type and composition (i.e . available glucose) to produce a spoilage prediction model . The method does not require sophisticated equipment, only a standard pH/ion meter, is cheap, needing only peptone and glucose, is relatively simple, and takes less than 2 h to perform. Clin Exp Immunol, 1994 Jun, 96(3), 466 - 9 Monoclonal IgM from patients with peripheral demyelinating neuropathies cross-react with bacterial polypeptides; Brouet JC et al.; Human monoclonal IgM associated with a demyelinating peripheral neuropathy often feature a distinct antibody activity directed against a glucuronyl sulphate epitope shared by myelin-associated glycoprotein (MAG), nerve glycolipids and low molecular weight peripheral nerve polypeptides . Earlier studies showed that these IgM use a diverse repertoire of VH and VL genes which exhibit somatic mutations, possibly indicative of an antigen-driven process . Here, we investigated whether such monoclonal IgM may react with environmental bacterial antigens . We found that six patients' sera and purified monoclonal IgM, as well as IgM from supernatants of three clonal anti-MAG-secreting cell lines reacted with unique 90-100 kD polypeptides from extracts of two out of 10 bacterial species . Purified MAG was able to inhibit this reactivity . These results indicate molecular mimicry as a possible mechanism of this immunomediated neuropathy and associated clonal lymphoid disease. Eur J Surg, 1994 Jun-Jul, 160(6-7), 357 - 62 Association between transfusion of stored blood and bacterial infective complications after biliary operations; Edna TH et al.; OBJECTIVE: To examine the association between blood transfusion and infective complications after biliary operations . DESIGN: Retrospective cohort study . SETTING: District hospital . SUBJECTS: 875 consecutive patients who required biliary operations . MAIN OUTCOME MEASURES: Postoperative infective morbidity in hospital . RESULTS: 73 patients (8%) developed postoperative infections in hospital . Univariate analysis showed that the development of infections was significantly associated with blood transfusion (p < 0.001), stones in the common bile duct (p < 0.001), operations on the common bile duct (p < 0.001), T-tube drainage (p < 0.001), duration of operation (p = 0.008), and age (p = 0.03) . Multivariate logistic regression analysis showed that only blood transfusion and stones in the common bile duct were independent predictors of infection . The corrected odds ratios for infection were 4.7 (95% confidence interval (CI) 2.4 to 9.3) when 1-3 units of blood were given and 5.6 (95% CI 2.3 to 13.6) when more than three units were given . CONCLUSION: Transfusion is an independent risk factor in the development of postoperative infection in hospital in patients who have had biliary operations. Eur J Surg, 1994 Jun-Jul, 160(6-7), 345 - 50 Thymopentin increases the survival of mice after allogeneic blood transfusion, bacterial gavage, and burn injury; Braga M et al.; OBJECTIVE: To investigate the effect of blood transfusion on mortality and the incidence of bacterial translocation in mice subjected to thermal burn or bacterial gavage, or both, and to assess the influence of thymopentin on mortality . DESIGN: Randomly controlled experiments . SETTING: University departments of surgery, immunology and nuclear medicine . MATERIAL: 235 Balb/c (H-2d) and C3H/HeJ (H-2k) mice . INTERVENTIONS: 8 groups of 20 mice each received: saline infusion (controls), blood transfusion (BT) alone, 20% burn alone, gavage with 1 x 10(10) Escherichia coli alone, BT and gavage, BT and burn, burn and gavage, or BT, burn, and gavage . A further 3 groups of 10 mice were all gavaged with 111In-biotin labelled E coli and randomised to additional BT and burn, BT alone, or burn alone . 98 mice that had had BT, burn, and gavage, were then randomised to receive thymopentin 0, 0.1, 1, or 5 mg/kg/day for 15 days . The impact of the pretreatment with thymopentin on PGE2 concentration was also evaluated in a separate group of 45 mice that received BT, burn, and gavage; or burn and gavage . MAIN OUTCOME MEASURES: Survival, degree of translocation . RESULTS: The highest mortality (75%) was in the BT, burn, and gavage group . BT alone significantly reduced survival in burned mice, whereas BT alone or associated with gavage had no effect . Thermal injury had the most influence on bacterial translocation, whereas BT did not increase it . Thymopentin significantly improved survival, particularly in the higher doses . The pretreatment with thymopentin significantly reduced PGE2 concentration after BT, burn and gavage . CONCLUSION: Burn injury significantly increased mortality in the presence of immune deficiency caused by BT . Thymopentin reduced mortality, possibly by immunomodulation. Am J Hosp Pharm, 1994 Jun 1, 51(11), 1429 - 32 Gentamicin pharmacokinetics in adults with bacterial endocarditis; Rosell-Rovira ML et al.; The pharmacokinetics of gentamicin in adult patients with endocarditis were studied . The records were reviewed for 64 patients treated for bacterial endocarditis and for whom serum gentamicin concentrations had been requested between May 1990 and May 1993 . The patients were divided into those with serum creatinine concentration (SCr) <1.2 mg/dL and those with SCr > or = 1.2 mg/dL . The measured serum gentamicin concentrations, patient demographic information, dosage interval, and SCr were entered into a pharmacokinetics program for analysis . The pharmacokinetic values evaluated were steady-state distribution volume (V), clearance (CL), elimination rate constant (k), and half-life (t1/2) . The mean +/- S.D . t1/2 and V of gentamicin were 4.7 +/- 2.4 hr and 0.29 +/- 0.11 L/kg . Half-life was correlated with SCr and V . These two variables may explain 66% of the variation in t1/2 . No difference in V was observed between patients with normal versus abnormal SCr . Men had a lower k than women . V and CL were lower in patients under age 60 than in older patients . Adult patients with endocarditis may have expanded gentamicin V . V and CL accounted for most of the variation in gentamicin t1/2. Mol Microbiol, 1994 Jun, 12(6), 993 - 1004 Bacterial binding protein-dependent permeases: characterization of distinctive signatures for functionally related integral cytoplasmic membrane proteins; Saurin W et al.; Bacterial binding protein-dependent transport systems belong to the superfamily of ABC transporters, which is widely distributed among living organisms . Their hydrophobic membrane proteins are the least characterized components . The primary structures of 61 integral membrane proteins from 35 uptake systems were compared in order to characterize a short conserved hydrophilic segment, with a consensus EAA---G---------I-LP, located approximately 100 residues from the C-terminus . Secondary structure predictions indicated that this conserved region might be formed by two amphipathic alpha-helices connected by a loop containing the invariant G residue . We classified the conserved motifs and found that membrane proteins from systems transporting structurally related substrates specifically display a greater number of identical residues in the conserved region . We determined a consensus for each class of membrane protein and showed that these can be considered as signatures. J Chir (Paris), 1994 Jun-Jul, 131(6-7), 291 - 5 {Hepatic tumoral form of bacterial infections . Diagnostic and therapeutic considerations apropos of 3 cases}; Berthet B et al.; When the clinical picture and complementary tests reveal a tumoural formation in the liver suggesting cancer, it is often difficult to diagnose such formations resulting from bacterial infection without surgical exploration . The diagnosis is particularly difficult since exclusive hepatic formations are extremely rare, the latency period is often long and specific markers are often lacking . We observed 3 such cases with local hepatic tumour-like formations due to tuberculosis, brucellosis and actinomycosis . Pathology examination of a liver biopsy can contribute to the diagnosis and avoid unnecessary exploratory laparotomy allowing successful medical treatment. Clin Infect Dis, 1994 Jun, 18(6), 958 - 62 Detection of culture-resistant bacterial pathogens by amplification and sequencing of ribosomal DNA; Wilson KH; Molecular phylogeny is profoundly influencing the field of bacterial evolution . New knowledge in this area has led to an exciting ability to detect and classify bacteria without culturing them . The process involved consists of either amplification or cloning of ribosomal DNA from a bacterial population, sequencing of this ribosomal DNA, and phylogenetic analysis of the sequences obtained . This approach has so far been applied successfully to four infectious diseases: bacillary angiomatosis, human ehrlichiosis, Whipple's disease, and Tyzzer's disease . Interpretation of data obtained by this method has been straightforward. Mutat Res, 1994 Jun, 312(3), 217 - 33 Recommendations for the performance of bacterial mutation assays; Gatehouse D et al.; At the International Workshop on the Standardisation of Genotoxicity Test Procedures, in Melbourne (27-28 February 1993), the current international guidelines for the correct conduct of bacterial mutation assays were considered, and the major differences between them were examined . An attempt was made to construct a scientifically based, internationally harmonized protocol . The main points of agreement were as follows . The consensus opinion was that there are currently insufficient data to justify a preference for either the preincubation or plate-incorporation methodologies as the initial test . Whichever method is used there was consensus agreement that the bacterial test battery should consist of S . typhimurium TA1537, TA1535, TA98 and TA100 . There was also consensus that the 3 strains TA97a, TA97 and TA1537 could be used interchangeably . Although it was not possible to achieve a consensus, the majority of the working group members agreed that strains for the detection of mutagens acting specifically on AT base pairs should be routinely included within the test battery . These strains may be S . typhimurium TA102 or E . coli WP2 strains (WP2 pKM101 and WP2 uvrA or WP2 uvrA pkM101) . With regard to study design it was universally agreed that 5 doses of test compound should be used in each experiment, and a majority agreement was obtained for 3 plates per dose . The use of 2 plates per dose is acceptable ONLY if the experiment is repeated . It is recommended that the negative controls may consist of solvent control alone provided that historical data are available to demonstrate lack of effect of the solvent in question . Positive control compounds should be included in all experiments, although the nature of these control compounds need not be specified in the guidelines . There was consensus agreement that for non-toxic freely soluble test agents, an upper limit of 5 mg/plate should be tested (5 microliters per plate for liquids) . For insoluble or toxic compounds, the recommendations were the same as those for other in vitro tests (see appropriate paper) . A consensus agreement was reached on the need to carry out further tests if equivocal results are obtained in the initial test, although it was generally agreed that the design of the repeat study should be left flexible . As there are little or no data to support the use of an exact repeat assay, a majority of the group recommended that negative results in the first test should be further investigated by either conducting a modified repeat (e.g . S9 titration) or by conducting the alternative methodology.(ABSTRACT TRUNCATED AT 400 WORDS) J Immunol, 1994 Jun 1, 152(11), 5468 - 76 IL-6 production by rat peritoneal mast cells is not necessarily preceded by histamine release and can be induced by bacterial lipopolysaccharide; Leal-Berumen I et al.; Mast cells produce a number of cytokines including IL-6 . In view of the large amounts of de novo synthesis induced by the activation of rat peritoneal mast cells and previous observations of expression of this cytokine by human lung mast cells, we have studied the regulation of IL-6 production . We examined the hypothesis that mast cell IL-6 production is not related to previous histamine release . Highly purified rat peritoneal mast cells were activated with anti-IgE, calcium ionophore A23187, or LPS . Histamine was used as a marker of preformed mediator release and IL-6 production was assessed by using the B9 hybridoma growth factor bioassay . Anti-IgE activation of rat peritoneal mast cells induced IL-6 production and histamine release . In contrast, LPS activation induced substantial, serum-dependent, IL-6 production without a significant level of histamine release . No preformed IL-6 was detected in the cells . Calcium ionophore induced histamine release from mast cells to a greater extent than did anti-IgE, but no A23187-induced IL-6 production was observed . A23187-treated cells retained high viability and produced a significant amount of TNF-alpha . To further examine the concordance of IL-6 production and histamine release we used mast cell stabilizing drugs . Dexamethasone and nedocromil significantly inhibited IL-6 production in response to anti-IgE . Our results demonstrate that there is not a direct relationship between mast cell degranulation and IL-6 production . Our observations are important for understanding the role of mast cells in inflammation and for developing strategies to modulate mast cell function in disease. Biochemistry, 1994 May 31, 33(21), 6564 - 70 The serine receptor of bacterial chemotaxis exhibits half-site saturation for serine binding; Lin LN et al.; Ligand binding to the serine receptor of Escherichia coli has been studied using isothermal titration calorimetry . Bacterial inner membranes enriched in the serine receptor (Tsr) were titrated as sonicated membrane samples and after solubilization in octyl beta-D-glucopyranoside (OG) to determine the number of moles of ligand bound per mole of receptor (n), the binding constant (Ka), and the enthalpy of binding (delta H) of serine to the receptor . The n value for serine binding to OG-solubilized Tsr protein (n = 0.5) was consistent with one molecule of serine binding to a receptor dimer, but in sonicated inner membrane samples, the n value was smaller (n approximately equal to 0.25), indicating that not all of the binding sites were accessible to added serine . At 7 and 27 degrees C, the values for Ka and delta H were equivalent for the membrane and OG-solubilized samples and were found to be 4.7 x 10(4) M-1 and -15 kcal/mol, and 3.6 x 10(4) M-1 and -18 kcal/mol, respectively . The influence of covalent modification at the sites of methylation on the affinity of the receptor for serine was also investigated, and found to have only a modest effect . The property of half-site saturation is suggestive of models for transmembrane signaling where the receptor subunit interactions are modulated by ligand binding. J Biol Chem, 1994 May 27, 269(21), 14951 - 6 The S12 ribosomal protein of Podospora anserina belongs to the S19 bacterial family and controls the mitochondrial genome integrity through cytoplasmic translation; Dequard-Chablat M et al.; In the filamentous fungus Podospora anserina, two nuclear genes are involved in the premature death syndrome associated with a site-specific deletion of the mitochondrial DNA: a mutant allele of the AS1 gene, encoding the cytoplasmic ribosomal protein S12, and an uncharacterized gene closely linked to the mating-type locus . We describe here the cloning and the sequencing of the wild-type and two mutant alleles of the AS1 gene . The P . anserina S12 protein belongs to the bacterial S19 ribosomal protein family and shows 72% identity with the S15 human ribosomal protein . Transformation experiments have shown that the AS1-4 mutation itself is responsible for the premature death phenotype and that it corresponds to a Gly to Asp change in the highly conserved COOH-terminal part of the protein . Use of antibodies directed against S12 did not permit detection of the mutant ribosomal protein inside the mitochondria . However, cross-reactions were observed with at least one mitochondrial ribosomal protein displaying a higher molecular weight than S12 . The mitochondrial protein does not seem to be a by-product of the AS1 gene but is more likely the mitochondrial homologue of S12 . These results strongly suggest that the mutant S12 protein acts indirectly to promote the mitochondrial deletion, via the cytoplasmic translation. Sci Total Environ, 1994 May 23, 146-147, 141 - 7 Inhibition of bacterial enzyme activity and luminescence by urban river sediments; Wei C et al.; The toxicological and ecological effects of pollutants in urban river sediments were studied . The sediments were chemically or physically fractionated, using selective extractants to separate the effects of metal and organic contaminants, and subsequently tested for the inhibition of bacterial enzyme activity and luminescence . In many cases the enzyme activity of the sediment-dwelling bacteria was inhibited by metals . The variations in inhibition were attributed to differences in sediment complexation of, rather than bacterial community tolerance to, metals . Non-polar organic compounds significantly increased the toxicity of urban river sediments, and it is proposed that polyaromatic hydrocarbons from storm-water are an important source of sediment toxicity. J Biol Chem, 1994 May 20, 269(20), 14553 - 8 Requirements of the secondary structures in the primary transcript for multicopy single-stranded DNA synthesis by reverse transcriptase from bacterial retron-Ec107; Shimada M et al.; Multicopy single-stranded DNA (msDNA) is produced by bacterial retroelements called retrons . It consists of single-stranded DNA that is linked to an internal G residue of an RNA molecule by a 2',5'-phosphodiester linkage . It has been demonstrated that specific primary sequences, as well as the secondary structures immediately downstream of the G residue, are essential for the cDNA priming reaction (Shimamoto, T., Hsu, M.-Y., Inouye, S., and Inouye, M . (1993) J . Biol . Chem . 268, 2684-2692) . We have now examined the requirement of the structures in the region corresponding to DNA for msDNA synthesis . The upper stem region consisting of 71 bases of msDNA-Ec107 was found not to be essential, and this region could be deleted to efficiently produce a truncated msDNA containing only a 36-base single-stranded DNA . Various mutations including base replacements, deletions, and insertions were constructed in the lower stem region . It was found that any mutations resulting in more stable secondary structures caused reduction in msDNA synthesis . The results indicated that reverse transcriptase requires a loose secondary structure in the template RNA near the cDNA priming site for cDNA elongation. Blood, 1994 May 15, 83(10), 2860 - 5 Hemoglobin enhances the production of tissue factor by endothelial cells in response to bacterial endotoxin; Roth RI; Human endothelial cells respond to bacterial endotoxin (lipopolysaccharide {LPS}) with changes that transform the endothelium into a surface with prominent procoagulant properties . Production of tissue factor (TF) in response to LPS is a major alteration that favors coagulation . Biologic activities of LPS have previously been shown to be enhanced by the presence of hemoglobin . Therefore, the ability of human hemoglobin (Hb) to modulate TF production by cultured human umbilical vein endothelial cells (HUVEC) was investigated . Cell-free Hb (10 mg/mL), either purified native (HbAo) or chemically cross-linked (alpha alpha Hb), was incubated with LPS (0.1 microgram/mL), and the mixtures then were added to HUVEC in culture . TF activity was quantified with a clotting assay and TF protein was measured with an enzyme-linked immunosorbent assay . Hb preparations greatly enhanced the production of TF activity (11- to 25-fold greater than TF produced by HUVEC alone) compared with minimal TF activity generated by LPS alone (only twofold greater than HUVEC alone) . The enhancement of LPS-induced TF activity was Hb concentration-dependent over a range of 1 to 100 mg/mL . Cross-linked alpha alpha Hb also greatly enhanced the production of TF protein compared with TF protein generated by LPS alone (12-fold greater v 3.5-fold greater than HUVEC alone, respectively) . The enhancement of LPS-induced TF protein was Hb concentration-dependent over a range of 0.1 to 2 mg/mL . Enhancement of TF activity by Hb required new protein synthesis . These results show that human Hb can augment the ability of LPS to induce endothelial cell TF and suggest that hemolysis associated with disseminated intravascular coagulation during sepsis may further stimulate coagulation . In addition, these results suggest a potential mechanism for generalized thrombosis in animals that has been associated with the infusion of cell-free Hb for resuscitation. J Immunol, 1994 May 15, 152(10), 4893 - 902 MHC-specific recognition of a bacterial superantigen by weakly reactive T cells; Surman S et al.; Previous studies have suggested that MHC class II polymorphism can influence the recognition of retroviral superantigen by murine T cells that have an intrinsically weak avidity for the superantigen . The aim of the present study was to determine whether bacterial superantigen recognition also is influenced by MHC polymorphism . Therefore, we screened for TCR with a low avidity for the bacterial superantigen SEB, and identified two V beta elements (V beta 14 and V beta 16) that had not been associated previously with SEB recognition . This finding extends the number of previously identified SEB-reactive V beta elements (V beta 6, V beta 7, V beta 8.1, V beta 8.2, and V beta 8.3) to at least seven . A detailed comparison of SEB recognition by V beta 14+ and V beta 8.2+ T cell hybridomas revealed two interesting features . First, SEB recognition by V beta 14+ hybridomas was relatively weak compared with V beta 8.2+ hybridomas . Second, in contrast to V beta 8.2+ hybridomas, individual V beta 14+ hybridomas responded differentially to SEB presented by either I-Ed or I-Ek molecules on the surface of L cell transfectants, indicating a role for polymorphic residues of the MHC in superantigen presentation . These findings demonstrate that T cell recognition of bacterial superantigens can be influenced by MHC polymorphism in a manner analogous to that of retroviral superantigen recognition, and that this characteristic is a feature of low avidity T cells . Taken together, these data support the hypothesis that there is a direct interaction between the TCR and MHC molecules during superantigen recognition. Proc Natl Acad Sci U S A, 1994 May 10, 91(10), 4249 - 53 Ribosome recycling factor (ribosome releasing factor) is essential for bacterial growth; Janosi L et al.; Ribosome releasing factor, product of the frr gene in Escherichia coli, is responsible for dissociation of ribosomes from mRNA after the termination of translation . It functions to "recycle" ribosomes and is renamed ribosome recycling factor in this paper . An E . coli strain was constructed (MC1061-2), which carried frame-shifted frr in the chromosome and wild-type frr on a temperature-sensitive plasmid . MC1061-2 is temperature-sensitive in its growth and does not segregate its frr-carrying plasmid under the plasmid incompatibility pressure . In contrast, isogenic E . coli carrying wild-type frr in the chromosome and mutated frr on the temperature-sensitive plasmid is not temperature-sensitive in its growth and segregates its plasmid from incompatible plasmids . All spontaneously formed thermoresistant colonies derived from MC1061-2 carried wild-type frr that resided either in the bacterial chromosome by re-exchange or in the plasmid, which became temperature-resistant . These observations establish that frr is an essential gene for cell growth. J Bone Joint Surg Am, 1994 May, 76(5), 664 - 6 Comparison of the results of bacterial cultures from multiple sites in chronic osteomyelitis of long bones . A prospective study; Patzakis MJ et al.; We evaluated the results of aerobic, anaerobic, and fungal cultures of specimens that had been obtained from multiple sites in thirty patients who had traumatic osteomyelitis with a sinus track . In each patient, we obtained specimens of material from the sinus track; specimens of purulent fluid, of soft tissue, and of bone obtained from curettage; and specimens from the bed of the involved bone . More than one organism grew on culture of the specimens from twenty-one of the patients; more than three organisms, from those of eleven patients; and ten organisms, from those of two patients . The same organisms grew on culture of the specimens from every site in only fourteen (47 per cent) of the thirty patients . We recommend that specimens of material from the sinus track; specimens of purulent material, of soft tissue, and of bone obtained from curettage; and specimens from the bed of the involved bone be obtained for culture before the treatment of chronic osteomyelitis with a draining sinus, so that as many of the infecting organisms as possible will be identified. J Immunol, 1994 May 1, 152(9), 4300 - 9 Control of the rat T cell response to retroviral and bacterial superantigens by class II MHC products and Tcrb-V8.2 alleles; Herrmann T et al.; The in vitro response of unprimed rat T cells to retroviral and bacterial superantigens (SAg) was analyzed with TCR V beta 8.2-, 8.5-, 10-, and 16-specific mAbs . Specific stimulation of V beta 8.2 and 8.5 CD4 cells was observed in the response to Mls1a, the retroviral SAg encoded by integrated provirus Mtv-7 (Mtv-7 SAg), which was presented by mouse B cells or mouse fibroblasts transfected with DR1 genes and the Mtv-7 SAg . Additionally, a strong response of V beta 16 CD4 cells to an as yet unidentified mouse SAg was found . Only some of the bacterial SAg known to stimulate mouse and human T cells also activated rat lymph node cells . SEA, SEE, and TSST-1 stimulated rat T cells well; SEB, SEC1, and SED did not . This defect was apparently a result of weak binding to rat MHC class II molecules because presentation by human MHC class II molecules restored T cell activation . Under these conditions, SEB stimulated V beta 8.2+ and 8.5+ CD4 and CD8 cells from Lewis rats . A comparison of several rat strains revealed an unresponsiveness to SEB or Mtv-7 SAg for V beta 8.2 cells from F344 and DA rats . Determination of the nucleotide sequences of the Tcrb-V8.2 of these strains revealed differences between SAg-responsive and SAg-unresponsive Tcrb-V8.2 in seven amino acids, four of them located in the putative SAg contact site . The significance of these findings for the evolution of TCR-SAg interactions is discussed. Biotechniques, 1994 May, 16(5), 932 - 7 A bacterial toxicity assay performed with microplates, microluminometry and Microtox reagent; Blaise C et al.; We have developed a procedure for undertaking a Microtox-based test by coupling microplate and microluminometric technologies . Sample dilutions are prepared in a 96-well polystyrene microplate kept at 15 degrees C, while the Microtox reagent and diluent are placed in an opaque, microluminometry-compatible 96-well microplate also kept at 15 degrees C . Exposure begins when sample aliquots are brought into contact with bacterial reagents in the opaque microplate . After specific exposure times (5, 15, 30 and 60 min), bacterial luminescence is rapidly measured by placing the opaque microplate in a microluminometer . Reproducibility of the procedure, as well as general agreement of EC50 end-point values with published reports, is demonstrated herein after toxicity trials with six metals (Ni2+, Cd2+, Zn2+, Pb2+, Cu2+ and Cr6+) . Results suggest that a 60-min exposure time may have value in getting more "sensitivity mileage" out of this Microtox-based assay . This microplate procedure possesses attractive features that augment sample throughput and information output . Further refinement and validation studies are ongoing in our laboratories. Am J Vet Res, 1994 May, 55(5), 654 - 9 Effects of dietary supplementation of fructo-oligosaccharides on small intestinal bacterial overgrowth in dogs; Willard MD et al.; Sixteen IgA-deficient German Shepherd Dogs with small intestinal bacterial overgrowth were randomized into 2 groups . One group was fed a chicken-based kibble diet; the other was fed the same diet, but with 1% fructo-oligosaccharides supplemented at the expense of cornstarch . After being exposed to the diets for 46 to 51 days, the group that ate the supplemented diet had significantly (P = 0.04) fewer aerobic/facultative anaerobic bacterial colony-forming units in fluid from the duodenum/proximal part of the jejunum, as well as in the duodenal mucosa . We could not detect significant differences in the species of bacteria found in the intestine of these 2 groups of dogs . We conclude that at least some dietary carbohydrates can affect small intestinal bacterial populations in dogs with small intestinal bacterial overgrowth. Mol Microbiol, 1994 May, 12(3), 351 - 7 Why are mammalian alkaline phosphatases much more active than bacterial alkaline phosphatases? Murphy JE, Kantrowitz ER. Mammalian alkaline phosphatases are 20-30-fold more active than the corresponding bacterial enzymes even though their amino acid sequences are 25-30% absolutely conserved . In the active-site region there are two noticeable differences between the sequences of the bacterial and mammalian enzymes . In the Escherichia coli enzyme positions 153 and 328 are Asp and Lys, respectively, but in the mammalian enzymes His is observed at both of these positions . Site-specific mutagenesis, genetic and X-ray crystallographic data, which will be summarized here, suggest that the His substitutions at positions 153 and 328 are primarily responsible for the differences in properties between the bacterial and mammalian alkaline phosphatases. J Perinatol, 1994 May-Jun, 14(3), 182 - 6 Pneumonia in the neonatal intensive care unit . Diagnosis by quantitative bacterial tracheal aspirate cultures; Ruderman JW et al.; Growth of > or = 10(5) colonies of bacteria per milliliter obtained at bronchoscopy in children and adults correlates with bacterial pneumonia . To determine whether quantitative tracheal aspirate cultures aid in diagnosis of pneumonia in the neonatal intensive care unit setting, tracheal aspirates were obtained from 25 infants who had recently undergone endotracheal intubation; 15 of the infants had suspected pneumonia and 10 control infants had undergone intubation for suspected apnea of prematurity (4 infants) or elective surgery (6 infants) . Studies also were performed to detect Mycoplasma, Ureaplasma, viruses, and Pneumocystis . Tracheal aspirates from 2 of 15 infants with suspected pneumonia grew > or = 10(5) bacteria, and 1 was positive for respiratory syncytial virus . These infants were considered to have pneumonia . In 12 infants whose tracheal aspirates grew < 10(5) bacteria, respiratory decompensation later was explained by other causes in 11 infants, and there was one false-negative culture . There were three false-positive tracheal aspirates in the control group . We conclude that tracheal aspirates of infants who have recently had an endotracheal tube placed may be useful for diagnosing pneumonia and for identifying the causative agent. Pediatr Neurol, 1994 May, 10(3), 259 - 61 Myelopathy secondary to neonatal bacterial meningitis; Coker SB et al.; Myelopathy is an infrequently reported complication of bacterial meningitis . Four patients with neonatal meningitis and cervical myelopathy are reported . This complication may be more frequent than presumed and should be closely assessed during evaluation . The conditions of most previously reported survivors improved or resolved and the majority involved the cervical spinal cord. Rev Med Interne, 1994 May, 15(6), 406 - 8 {Cerebrovascular complications of non-bacterial thrombotic endocarditis: value of transesophageal echocardiography}; Berger E et al.; Non bacterial thrombotic endocarditis is a frequent cause of cerebrovascular complications in patients with cancer . Clinical features are often misleading and symptoms are hardly linked to valvular thrombosis in the absence of systemic embolism . We describe here a case mainly illustrating two aspects: clinical and neuroradiological manifestations of multiple distal brain infarcts and the value of transoesophageal echocardiography for the diagnosis. J Clin Periodontol, 1994 May, 21(5), 347 - 50 Evaluation of a new toothbrush concept with regard to bacterial elimination . Imprint study using scanning electron microscopy; Apiou J et al.; The efficacy of a new toothbrush filament layout concept (Topix, Peridental, France) was compared to that of a standard vertical-tuft toothbrush . Bacterial and exogenous deposit elimination were used as parameters of efficacy . 30 dental surgery students took part in the study . Plaque index scores were calculated according to a pre-defined protocol . Imprints of the 6 anterior teeth were taken before and after brushing with the 2 types of brushes, without toothpaste or rinsing . Imprints were examined by scanning electron microscopy (SEM) . 12 h after brushing, imprint examination revealed bacterial flora polymorphism and the amount of dental plaque accumulated at the cervical third zone of teeth . Automated quantification in this zone of exogenous bodies showed that after brushing with vertical-tuft and cross-tuft brushes, there remained 1.26 mm2 and 0.83 mm2 of dental plaque, squamae, and blood residues, respectively . The plaque index values correlated to scanning electron microscopic observations . There was no significant difference in terms of efficacy between the cross-tuft and vertical-tuft toothbrushes. Br J Urol, 1994 May, 73(5), 508 - 15 Lymphocyte sub-populations in the bladder wall in normal bladder, bacterial cystitis and interstitial cystitis; Christmas TJ; OBJECTIVE: To identify lymphocyte sub-populations within the bladder wall in normal controls, in patients with bacterial cystitis (BC) or interstitial cystitis (IC) . PATIENTS AND METHODS: Bladder biopsies were taken from 21 patients with IC, four with BC and five normal controls . Immunofluorescent staining of biopsies was performed using monoclonal antibodies to the T cell markers CD3, CD4, CD8, TCR delta 1, delta TCS1, beta F1 and IML . Plasma cells were identified using monoclonal antibodies to IgA, IgG and IgM . The location and numbers of cells were determined . Peripheral blood lymphocyte counts were also performed in each case . RESULTS: T cells (CD4+ and CD8+) were significantly more abundant within the bladder biopsies from patients with interstitial cystitis than in those from the normal controls, and were present within the urothelium and submucosa but not the detrusor . There was no difference between the numbers of T cells present in the biopsies from patients with interstitial cystitis and in those with bacterial cystitis . However, the presence of gamma delta T cells was significantly associated with interstitial cystitis . IgA+ and IgM+ plasma cells were located within the urothelium and submucosa but not the detrusor in patients with IC and the numbers present in these patients were significantly greater than in those with BC or the normal controls . The peripheral blood lymphocyte counts were not altered in any of the study cases . CONCLUSION: The increased numbers of CD4+, CD8+ and gamma delta cells as well as IgA+, IgG+ and IgM+ plasma cells within the urothelium and submucosa in patients with IC suggest that these cells play an active role in the pathogenesis of the disorder . The previously described abnormal urothelial expression of HLA-DR and hyperexpression of Class I molecules in patients with IC may enable these cells to cause destruction of the urothelium--the end stage of IC. Biophys Chem, 1994 May, 50(1-2), 191 - 201 Comparative studies on ion pumps of the bacterial rhodopsin family; Mukohata Y; Bacteriorhodopsin (proton pump), halorhodopsin (anion pump), sensory rhodopsin and phoborhodopsin (photosensors) are found in Halobacterium salinarium (halobium) . In some other strains, other sets of rhodopsin pumps and sensors have been found . Here, these bacterial rhodopsins are classified according to their amino acid sequence homologies, and their host genera are assigned on the basis of 16S rRNA sequence comparison . Haloarcula is the host for cruxrhodopsins and a new genus (temporarily "Halorubra") is the host for archaerhodopsins . Difference in the all-trans:13-cis ratios of retinal in two proton pumps (bacteriorhodopsin and archaerhodopsin-2) at equilibrium states in the dark was ascribed to only one amino acid residue in the retinal pocket . This predicted methionine-145 in bacteriorhodopsin was point-mutated to phenylalanine as in archaerhodopsin-2 . The mutated bacteriorhodopsin (M145F) became to show the same dark-adapted isomer ratio that archaerhodopsin-2 shows . Chimeric proton pumps were made by exchanging genes of one or more helix regions of two similar pumps (archaerhodopsin-1 and -2) in order to know structural delicacy of the inter-helix space . Preliminary results show that some photochemical properties depend on one helix or one distinct amino acid residue on the helix . Such new lines initiated by our archaerhodopsins are discussed for studying structure and function of these unique bacterial rhodopsins. J Biolumin Chemilumin, 1994 May-Jun, 9(3), 185 - 200 Low-light image analysis of transgenic organisms using bacterial luciferase as a marker; Langridge W et al.; Methods for measurement of a novel light-emitting reporter gene system in bacteria, yeast, plant cells, plant tissues and intact plant organs are described . The principle underlying the assay procedures is the bacterial luciferase catalysed oxidation of reduced flavin mononucleotide (FMNH2) in the presence of the ten carbon aldehyde decanal, to yield FMN, decanoic acid, water and a photon of light at 490 nm which can be captured by X-ray film, a photomultiplier tube or, for in vivo measurements, an image-intensifier coupled to a video camera . This light measuring assay system is sensitive, easy to use, inexpensive, does not require radioactivity, and has been used successfully for rapid detection of bacterial transformants, the quantitative measurement of transient and stable gene expression in bacteria and yeast, and in vivo measurement of temporal and spatial gene expression throughout plant and animal development. Mol Biochem Parasitol, 1994 May, 65(1), 39 - 49 Autonomous replication of bacterial DNA plasmid oligomers in Leishmania; Papadopoulou B et al.; Extrachromosomal amplicons are frequently observed in drug-resistant Leishmania . A dominant selectable marker, the neomycin phosphotransferase gene, was introduced by gene targeting in a circular amplicon derived from the H locus of Leishmania in a mutant cell . This recombinant amplicon was isolated and transfected in a wild-type cell . The amplicon was kept in the wild-type cells, provided the selective pressure was maintained, suggesting that it was capable of autonomous replication . Novel Leishmania expression vectors suited for stable transfections were made to isolate, by a high transformation assay, the putative origin of replication in the amplicons . However, these plasmids, which did not contain a single Leishmania nucleotide, were found as extrachromosomal circular oligomers in Leishmania transfectants . Their relative stability, in addition to changes in their methylation pattern, indicated that these plasmids were most likely replicating . No specific sequences seem to be required for replication (and expression) in Leishmania, therefore precluding the isolation of origins of replication by genetic transformation. Int J Immunopharmacol, 1994 May-Jun, 16(5-6), 469 - 73 Immunomodulation by bacterial fractions; Barot-Ciorbaru R; The cells of Nocardia opaca are a source of potent immunostimulating substances, differing in solubility and in the presence or absence of peptidoglycan . Three classes of N . opaca fractions have been investigated: (1) NDCM (Nocardia delipidated cell mitogen), the starting fraction; (2) PG (peptidoglycan)-containing fractions: CW (cell walls), PG, and fractions containing soluble PG derivatives such as NWSM (Nocardia water soluble mitogen) and NSPD (Nocardia soluble peptidoglycan derivative); and (3) soluble fractions devoid of PG, such as NWSMP (NWSP-pellet) and CyI (cytoplasmic membrane) . This report summarizes our results on the relationship between the structure of N . opaca fractions and their stimulatory activities on mouse, rabbit, germ-free piglet and human lymphocytes and monocytes/macrophages . The studies of biological properties of these compounds are the result of an extensive collaborative effort of several teams. J Med Assoc Thai, 1994 May, 77(5), 266 - 70 Significance of ascitic fluid white blood cells, pH, lactate, and other chemistry in immediate diagnosis of spontaneous bacterial peritonitis; Neungton N et al.; From 1989 to 1991, 68 cirrhotic patients, 47 with uninfected ascites and 21 with SBP were studied for the significance of ascitic fluid pH, lactate, PMN count and other chemistry for immediate diagnosis of SBP . It was revealed that ascitic fluid PMN count if over 500 per mm3, the increased lactate, or decreased glucose level, strongly supported the diagnosis of SBP . In cases of suspecting SBP but with low PMN count the ascitic values of lactate, glucose and pH will guide the diagnosis . If the ascitic lactate plus glucose, or lactate plus pH are above the cut off levels (lactate > 25 mg/dl; glucose < 60 mg/dl and pH < 7.35) the diagnosis is strongly suggestive . The ascitic fluid pH and A-AF pH gradient were not of diagnostic value due to instability of pH after tapping . For other chemistry in the ascitic fluid, there was a slight increase in ADA level in SBP, but for glucose, protein and glutamine levels, there was no difference among the groups with and without SBP. J Burn Care Rehabil, 1994 May-Jun, 15(3), 207 - 12 The Caco-2 cell monolayer system as an in vitro model for studying bacterial-enterocyte interactions and bacterial translocation; Cruz N et al.; Partly because of inherent limitations of in vivo models, the cellular mechanisms underlying the process of bacterial translocation across the intestinal epithelial barrier are incompletely understood . We therefore used the Caco-2 intestinal cell line as an in vitro model to examine the bacterial translocation process under controlled conditions . Caco-2 cells were grown on porous membranes in the upper compartment of a two-compartment system . Caco-2 cells were cultured for 7, 14, 21, or 28 days . Cellular confluence and tight junction integrity were verified by measurements of dextran permeability and transepithelial electrical resistance . Bacterial translocation was measured by culturing the bacteria (E . coli C25) that were able to cross the Caco-2 cell monolayer . The passage of E . coli C-25 and dextran across the Caco-2 monolayer was higher and the transepithelial electrical resistance lower after 7 days of culture than after 14 or 21 days of culture . The Caco-2 cells became impermeable to dextran blue after 14 days of culture with an average transepithelial electrical resistance of 173.1 +/- 9.24 ohms.cm2 . When increasing doses of (10(2)-10(9) colony-forming units) of E . coli were tested in 14-day-old Caco-2 monolayers, bacterial translocation occurred in a time- and dose-dependent fashion . Once cellular confluence and tight junction integrity have been established, bacterial translocation across Caco-2 cells appears to be a time- and dose-dependent process. Biochem Biophys Res Commun, 1994 Apr 29, 200(2), 908 - 15 On the structure of mitochondrial porins and its homologies with bacterial porins; Rauch G et al.; By use of computer modelling, we have predicted a model of 16 transmembrane beta-strands for mitochondrial porins structure from human, Saccharomyces cerevisiae, Neurospora crassa and Dictyostelium discoideum . The proposed model takes into account biochemical and immunological data reported in the literature, as well as electrophysiological results obtained with yeast mitochondrial porins with mutations at selected amino acids . The predicted structure is very similar to that of some bacterial porins, as apparent from the homology of their hydropathic profiles. J Mol Biol, 1994 Apr 29, 238(2), 187 - 98 pIV, a filamentous phage protein that mediates phage export across the bacterial cell envelope, forms a multimer; Kazmierczak BI et al.; Filamentous phage pIV is an outer membrane protein required for phage assembly and secretion . Chemical cross-linking and sedimentation experiments have been used to demonstrate that pIV from f1-infected Escherichia coli exists as a homo-multimer, probably composed of 10 to 12 subunits . pIV secreted from spheroplasts remains soluble and does not form multimers . Synthesis of pIV from distantly related filamentous phages or from a bacterial homolog that participates in a specialized form of extra-cellular protein secretion in the same cell with pIVf1 resulted in the formation of mixed multimers . This suggests that the homologous proteins themselves form homo-multimers . These structures could form gated channels that conduct assembling phage or specific substrate proteins across the outer membrane to the extracellular milieu. Proc Natl Acad Sci U S A, 1994 Apr 26, 91(9), 3867 - 71 Redox modulation of the expression of bacterial genes encoding cysteine-rich proteins in plant protoplasts; Pineiro M et al.; Activity of neomycin phosphotransferase II (NPTII; gene, neo; five cysteines) in tobacco protoplasts transfected with fusions of the octopine TR2' or cauliflower mosaic virus 35S promoter and the neo gene, with or without a signal peptide, increased up to 8-fold in response to externally added dithiothreitol at concentrations that did not affect protoplast viability (up to 2.5 mM) . Activity of phosphinothricin acetyltransferase (PAT; gene, bar; one cysteine) expressed under control of the TR1' or 35S promoter was not similarly affected, thus excluding a redox modulation of transcription as the mechanism of NPTII activation by dithiothreitol . Western-blot analyses showed an increase in the amount of protein in response to dithiothreitol, whereas neither the steady-state level of NPTII mRNA nor the specific activity of the purified enzyme was affected . The same type of modulation was observed for transiently expressed beta-glucuronidase (nine cysteines) produced from a fusion with the 35S promoter, with or without a signal peptide . Limitation of cotranslational and/or early posttranslational steps by excessively oxidizing sulfhydryl/disulfide redox potentials is postulated to explain the low net accumulation of cysteine-rich proteins of bacterial origin (i.e., NPTII and beta-glucuronidase) when expressed in plant protoplasts, and the marked increase in such proteins in response to externally added dithiothreitol. Biochemistry, 1994 Apr 26, 33(16), 4953 - 65 The binding sites of quinones in photosynthetic bacterial reaction centers investigated by light-induced FTIR difference spectroscopy: assignment of the QA vibrations in Rhodobacter sphaeroides using 18O- or 13C-labeled ubiquinone and vitamin K1; Breton J et al.; Light-induced FTIR difference spectra of the photoreduction of the primary quinone acceptor QA have been obtained for Rhodobacter sphaeroides RCs reconstituted with a series of isotopically labeled quinones in order to separate the contributions of the quinone from those of the protein . The isotopic shifts observed in the QA-/QA spectra of RCs reconstituted with ubiquinones (Q1, Q6) or vitamin K1 18O-labeled on their carbonyl oxygens and with fully 13C-labeled Q8 lead to a clear identification of the quinone bands from both the neutral and anion forms . Double-difference spectra from pairs of QA-/QA spectra obtained from 18O/16O Q6, 18O/16O Q1, 13C/12C Q8, 13C18O/12C16O Q8, and 18O/16O vitamin K1 allow the C = O modes of QA in vivo to be identified unambiguously for the first time . For all the investigated unlabeled quinones, two carbonyl bands are demasked, at 1660 and 1628 cm-1 for neutral ubiquinones and at 1651 and 1640 cm-1 for vitamin K1, while C = C bands are found at 1608 and 1588 cm-1 for vitamin K1 and at 1601 cm-1 for ubiquinones . Compared with the spectra of the isolated quinones, the generally smaller width observed for the C = O and C = C bands in vivo suggests precise interactions between the quinone and the contours of the protein at a single, well-defined QA site . The different frequency downshifts of the two C = O bands upon binding to the QA site underscore the inequivalence of the two carbonyls in providing asymmetrical bonding interactions with the protein . The comparison of the isotopic shifts observed for the various quinone C = O and C = C bands in vitro and in vivo demonstrates that the admixture of C = O and C = C characters in these modes is strongly affected by the binding of QA to its anchoring site . In particular, the bands at 1628 and 1601 cm-1 of Q6 in vivo exhibit highly mixed C = O and C = C characters . In contrast, the methoxy groups of the ubiquinones do not appear to suffer large strain upon binding . The closeness of the QA-/QA spectra for Q1 and Q6 indicates that a possible role of the chain in providing the proper positioning of the quinone ring in the site for both the oxidized and reduced states of QA cannot extend significantly beyond the first isoprene unit.(ABSTRACT TRUNCATED AT 400 WORDS) EMBO J, 1994 Apr 15, 13(8), 1998 - 2006 A mitochondrial homolog of bacterial GrpE interacts with mitochondrial hsp70 and is essential for viability; Bolliger L et al.; Mitochondrial hsp70 (mhsp70) is located in the matrix and an essential component of the mitochondrial protein import system . To study the function of mhsp70 and to identify possible partner proteins we constructed a yeast strain in which all mhsp70 molecules carry a C-terminal hexa-histidine tag . The tagged mhsp70 appears to be functional in vivo . When an ATP depleted mitochondrial extract was incubated with a nickel-derivatized affinity resin, the resin bound not only mhsp70, but also a 23 kDa protein . This protein was dissociated from mhsp70 by ATP . ADP and GTP were much less effective in promoting dissociation whereas CTP and TTP were inactive . We cloned the gene encoding the 23 kDa protein . This gene, termed GRPE, encodes a 228 residue protein, whose sequence closely resembles that of the bacterial GrpE protein . Microsequencing the purified 23 kDa protein established it as the product of the yeast GRPE gene . Yeast GrpEp is made as a precursor that is cleaved upon import into isolated mitochondria . GrpEp is essential for viability . We suggest that this protein interacts with mhsp70 in a manner analogous to that of GrpE with DnaK of E.coli. Gene, 1994 Apr 8, 141(1), 79 - 84 A general vector, pASK84, for cloning, bacterial production, and single-step purification of antibody Fab fragments; Skerra A; The expression vector pASK84 was designed for the convenient cloning of immunoglobulin variable domain genes, as well as periplasmic secretion of the corresponding F(ab) fragment in Escherichia coli . The plasmid provides the constant domain genes of mouse IgG1/kappa with a hexa-histidine tag fused to the C terminus of the heavy chain . This strategy enables the rapid and efficient purification of the functional recombinant F(ab) fragment via immobilized metal affinity chromatography . The versatility of this expression and purification system is demonstrated using the variable domains of the well-characterized anti-lysozyme antibody D1.3. Genitourin Med, 1994 Apr, 70(2), 121 - 3 Treatment of bacterial vaginosis with a three day course of 2% clindamycin vaginal cream: a pilot study; Dhar J et al.; OBJECTIVE--To evaluate the efficacy and safety of a 3 day course of 2% clindamycin cream in the treatment of bacterial vaginosis . DESIGN--A prospective, randomised, double blind, placebo controlled study . SETTING--Department of Genitourinary Medicine, Royal Liverpool University Hospital . SUBJECTS--55 female patients aged 18 years and over, and premenopausal, who spontaneously or after questioning complained of symptoms of bacterial vaginosis . RESULTS--55 patients were enrolled . 44 patients were evaluable at Visit 1 when among the 23 who received clindamycin cream bacterial vaginosis was not present in 22 (95.6%) and only one failed treatment . Of the 21 patients in the placebo group only one (4.8%) patient was cured and 20 (95.2%) were failures . Of the 17 patients evaluable at Visit 2 in the clindamycin group, bacterial vaginosis was not present in 14 (82.4%) and had recurred in three . No serious adverse events were noted in either group . CONCLUSION--This pilot study provides encouraging evidence of the efficacy and safety of a 3 day course of 2% clindamycin cream in bacterial vaginosis. Br J Surg, 1994 Apr, 81(4), 579 - 84 Bacterial translocation, intestinal ultrastructure and cell membrane permeability early after major liver resection in the rat; Wang XD et al.; The process and route of bacterial translocation from the gut after major liver resection remain unclear . In the present study enteric bacterial translocation, enterocyte ultrastructure in the ileum and colon, the process and route of bacterial invasion and the permeability of the cell membrane system and blood-tissue barrier were evaluated in rats receiving sham operation, and 70 or 90 per cent hepatectomy . The incidence of bacterial translocation to mesenteric lymph nodes was 80-100 per cent in rats 6 h after 70 per cent and 2-4 h after 90 per cent hepatectomy, and 80-100 per cent to the systemic circulation 2-4 h after 90 per cent hepatectomy but only 20 per cent to the portal vein . An increase in bacterial adherence to the intestinal surface, damage to the permeability of the cell membrane system and blood-tissue barrier, and pathological alterations in the ileum and colon developed, correlating with the extent of liver removed and the time that had passed after hepatectomy . Most translocating bacteria appeared in morphologically intact enterocytes with increased membrane permeability, in antigen-presenting cells and in submucosal lymphatics, but some bacteria were also seen within damaged enterocytes 4h after 90 per cent hepatectomy . These results indicate that altered permeability of the cell membrane system may be one of the earliest characteristics of challenged enterocytes, and that enteric bacteria translocate through both morphologically normal and abnormal enterocytes . Translocation occurred mainly into the lymphatics, bacteria either being 'carried' by antigen-presenting cells or entering by active invasion. J Paediatr Child Health, 1994 Apr, 30(2), 160 - 4 Lumbar punctures in suspected bacterial meningitis: too many or too few? Selby A, Isaacs D, Gillis J, Hanson R, O'Connell A, Schell D, Van Mai T. Children aged 1 month to 14 years admitted to the Royal Alexandria Hospital for Children during a 10 month period with suspected meningitis were studied prospectively . The aims were to determine how often lumbar puncture (LP) was delayed or never done, in relation to the outcome of all children, in order to determine the risks of LP and the risks of not doing LP . Of 218 children with suspected meningitis, LP was performed immediately in 195 (89.4%) . Meningitis was diagnosed in 49 of these (bacterial 18, viral 31) . No child developed cerebral herniation due to immediate LP . There were 11 traumatic taps and two children required repeated attempts . Lumbar puncture was delayed, but performed at a later time in 17 children, of whom three had proven bacterial meningitis, 1 had presumed bacterial meningitis but no organism was detected and 13 had alternative diagnoses . Six children never had an LP, although ventricular cerebrospinal fluid was obtained from two . Four of these six children had presumptive bacterial meningitis, one had tuberculous meningitis presenting with acute hydrocephalus and diagnosed post-mortem, and one had a very poor neurological outcome and no final diagnosis was reached . Of the 27 children with bacterial meningitis, LP was performed immediately in 18, or two-thirds . There were only minor adverse effects of immediate LP . Delayed LP probably resulted in failure to identify the organism in one child with bacterial meningitis, but did not adversely affect outcome in any child . Of the six children in whom LP was never performed, in only one was no final diagnosis reached.(ABSTRACT TRUNCATED AT 250 WORDS) Arch Dis Child, 1994 Apr, 70(4), 327 - 30 Bacterial contamination of enteral feeds; Patchell CJ et al.; Enteral nutrition is increasingly used to provide nutritional support for children in hospital and at home . No suitable formula is available for preschool children, however, and until recently a modular feed has been prepared . The hypotheses were examined that the use of a modular feed is associated with increased bacterial contamination, and that contamination is more common in the home than in hospital . Thirty five children receiving enteral nutrition initially in hospital and subsequently at home were allocated randomly to receive either a modular feed or a newly available sterile ready to use paediatric feed . Samples of feed were taken from the nutrient container immediately after filling and at the end of feeding . The results show that feed contamination is common in hospital and at home, but significantly more so at home . The data indicate the importance of hygiene training for parents and the desirability of a ready to use formula. Gut, 1994 Apr, 35(4), 455 - 60 Effect of omeprazole on intragastric bacterial counts, nitrates, nitrites, and N-nitroso compounds; Verdu E et al.; Previous studies have suggested that profound inhibition of gastric acid secretion may increase exposure to potentially carcinogenic N-nitroso compounds . The aim of this study was to find out if the proton pump inhibitor omeprazole (20 mg daily) is associated with increased concentrations of potentially carcinogenic N-nitroso compounds in gastric juice . The volume of gastric contents, number of bacteria, and concentrations of nitrates, nitrites, and N-nitroso compounds was determined in gastric aspirates obtained after an overnight fast in 14 healthy volunteers (7M:7F) after one week of treatment with placebo, and one and two weeks' treatment with omeprazole . Median bacterial concentrations were 1.0 x 10(4) (range 5.0 x 10(3)-5.0 x 10(6)) colony forming units (CFU)/ml after one weeks' treatment with placebo and increased significantly to 4.0 x 10(5) (0-3.3 x 10(7)) CFU/ml after two weeks' treatment with omeprazole (p < 0.05) . A similar increase was seen in the concentration of nitrate reducing bacteria . There was no difference in the volume of gastric aspirates after treatment with omeprazole when compared with placebo (65 (29-155) ml v 42 (19-194) ml) . The concentration of N-nitroso compounds was 0.13 (0-1.0) mumol/l after two weeks of omeprazole, which was not significantly different from that seen with placebo (0.15 (0-0.61) mumol/l) . There was also no increase in the concentrations of nitrates or nitrites . It is concluded that omeprazole (20 mg once daily) for two weeks in healthy volunteers is associated with gastric bacterial proliferation but does not increase concentrations of N-nitroso compounds. J Gen Virol, 1994 Apr, 75 ( Pt 4), 733 - 41 Characterization of the herpes simplex virus type 1 strain 17+ neurovirulence gene RL1 and its expression in a bacterial system; McKie EA et al.; The DNA sequence of herpes simplex virus type 1 (HSV-1) strain 17+ in the region coding for the polypeptide ICP34.5 predicts a protein of 248 amino acids with a proposed M(r) of 26,158 . The entire RL1 open reading frame was cloned into the expression vector pET8c to enable over-expression of ICP34.5 in Escherichia coli . The expressed protein was partially purified and used as an immunogen to produce a polyclonal antiserum in rabbits . Construction of an ICP34.5 null mutant (1771), demonstrated that the predicted open reading frame for ICP34.5 in strain 17+ is correct and confirmed that HSV-1 strain 17+ ICP34.5 specifically determines neurovirulence . The specificity of the anti-serum directed against the E . coli-expressed ICP34.5 was defined by Western blotting of wild-type and RL1-negative infected cell extracts. J Biol Chem, 1994 Apr 1, 269(13), 9429 - 35 Interaction of a legume lectin with two components of the bacterial cell wall . A crystallographic study; Bourne Y et al.; We describe herein the refined high resolution x-ray structures of two components of the bacterial cell wall, muramic acid and muramyl dipeptide complexed to isolectin I from Lathyrus ochrus seeds . In both complexes, only the ring hydroxyl oxygen atoms of the bound sugar establish direct hydrogen bonds with isolectin I, as in the case of all the previously determined monosaccharide-lectin complexes . In addition, the lactyl methyl of both components strongly interacts via hydrophobic contacts with the side chains of residues Tyr100 and Trp128 of isolectin I, which could explain the higher affinity of isolectin I for muramic acid as compared with glucose . These 2 residues, however, are not involved in the stabilization of the oligosaccharide-isolectin I complexes . The dipeptide (D-Ala-D-iGln) of the second component is in stacking interaction with the N-acetyl group of glucose and with loop Gly97-Gly98 of isolectin I . In addition to these van der Waals' contacts, the dipeptide interacts with the lectin via well ordered water molecules also . Superposition of the structures of the muramyl dipeptide complex and of the muramic acid complex shows that the glucose ring in the dipeptide compound is tilted by about 15 degrees in comparison with that of muramic acid . The fact that the lactyl group has the same confrontation in both components reveals that the lectin is stereospecific and recognizes only diastereoisomer S of this group, which better fits the saccharide-binding site. Crit Care Med, 1994 Apr, 22(4), 690 - 6 Bacterial translocation in burned mice after administration of various diets including fiber- and glutamine-enriched enteral formulas; Zapata-Sirvent RL et al.; OBJECTIVE: Severe burn injury can produce acute gastrointestinal derangements which may facilitate bacterial translocation to mesenteric lymph nodes . We studied the effects of feeding different dietary formulations on bacterial translocation in burned mice . DESIGN: Prospective, blinded, nonrandomized laboratory study . SETTING: Research laboratory . SUBJECTS: One hundred sixty-nine female, outbred, CF-1 mice, 8 to 12 wks of age . INTERVENTIONS: Anesthetized mice received a 32% total body surface area, full-thickness burn injury . Mice were then fed with: a) mouse chow; b) a low-residue enteral formula; c) a high-protein, high-fat enteral formula; d) an enteral formula with high concentrations of supplemental glutamine; or e) an enteral formula that contains soy fiber . MEASUREMENTS AND MAIN RESULTS: Burned mice that were fed the low-residue enteral formula demonstrated increased mortality rate (21.2%, p = .05) compared with chow-fed mice in the 2-day postburn period (0 mortality); other burn-diet groups had intermediate mortality rates . In surviving mice, bacterial translocation was found to be: a) lowest in the group fed chow (31.0%) and the high glutamine formula (30.8%); b) intermediate in the group fed formula and soy fiber (44.8%, NS compared with burn-chow group); and c) highest in the group receiving the low-residue enteral formula (73.1%, p < .005) and high-protein, high-fat enteral formula (59.3%, p < .05) . CONCLUSIONS: Dietary composition markedly affects bacterial translocation in this animal burn model . Commercial enteral diets containing fiber and high concentrations of glutamine provide protection for the gut after burn injury and reduce the occurrence of bacterial translocation in this animal model. Hepatology, 1994 Apr, 19(4), 1029 - 33 Enzyme inhibitory autoantibodies to pyruvate dehydrogenase complex in primary biliary cirrhosis differ for mammalian, yeast and bacterial enzymes: implications for molecular mimicry; Teoh KL et al.; Primary biliary cirrhosis is a chronic autoimmune disease in which serum autoantibodies against the mitochondrial 2-oxo acid dehydrogenase enzyme complexes (M2 antibodies) are regularly present . Molecular mimicry of host proteins by bacterial counterparts is a suggested explanation for the origin of these autoantibodies . We tested this hypothesis by measuring the functional reactivity of serum autoantibodies by means of an enzyme inhibition assay against pyruvate dehydrogenase complex from different sources: mammalian, Saccharomyces cerevisiae and Escherichia coli . The 10 primary biliary cirrhosis sera all reacted on immunofluorescence study for M2 antibodies and on immunoblotting with the pyruvate dehydrogenase complex E2 subunit from each of the three enzymes, but there were strikingly different inhibitory capacities . The primary biliary cirrhosis sera were highly inhibitory for mammalian pyruvate dehydrogenase complex (10 of 10 inhibitory; mean level of inhibition, 99%), moderately inhibitory for yeast pyruvate dehydrogenase complex (10 of 10 inhibitory; mean level, 70%) and weakly inhibitory for Escherichia coli pyruvate dehydrogenase complex (4 of 10 inhibitory; mean level, 26%) . Thus, with a functional assay that depends on epitope recognition of primary biliary cirrhosis sera, cross-reactivity between mammalian and bacterial pyruvate dehydrogenase complex enzymes is low and molecular mimicry, at least at the B-lymphocyte level, is not supported. Plant Cell Physiol, 1994 Apr, 35(3), 425 - 37 cDNA cloning of monodehydroascorbate radical reductase from cucumber: a high degree of homology in terms of amino acid sequence between this enzyme and bacterial flavoenzymes; Sano S et al.; The FAD-enzyme monodehydroascorbate (MDA) reductase catalyzes the regeneration of ascorbate from the MDA radical using NAD(P)H as the electron donor {Hossain and Asada (1985) J . Biol . Chem . 260: 12920} . We cloned a cDNA of MDA reductase from cucumber seedlings and deduced its entire sequence of amino acid residues . The cDNA library from cucumber seedlings in the expression vector was screened with an antiserum against cucumber MDA reductase . Inserts from three immunoscreened clones hybridized with two oligonucleotide probes designed on the basis of the sequences of two peptide fragments from the cucumber enzyme . The nucleotide sequences of these three clones were determined and the longest one contained an open reading frame of 1,302 bp in length . The molecular mass of the translation product predicted from the open reading frame was 47 kDa, the same as that determined for the purified enzyme . The amino acid sequences determined from fragments of lysyl endopeptidase-digested MDA reductase could be aligned with that deduced from the open reading frame, although substitution of several residues was apparent . Thus, the open reading frame encoded an isozyme of MDA reductase of cucumber different from the purified enzyme . MDA reductase has the FAD- and NAD(P)H-binding domains of flavoproteins but shares only limited homology in terms of amino acid sequence with flavoenzymes from eukaryotes. Blood Coagul Fibrinolysis, 1994 Apr, 5(2), 211 - 20 Expression of tissue factor and tissue factor pathway inhibitor in monocytes in response to bacterial lipopolysaccharide and phorbolester; van der Logt CP et al.; The monocyte is the only circulating cell type capable of initiating blood coagulation by the expression of tissue factor (TF) . The mechanism and kinetics of TF mRNA and TF activity induction in human peripheral blood monocytes (HPBM) in response to bacterial lipopolysaccharide (LPS) and phorbol myristate acetate (PMA) were investigated . Northern blot analysis showed that both LPS and PMA induce a transient accumulation of TF mRNA in HPBM, that reaches maximum levels after 3-6 h and rapidly declines thereafter . Nuclear run-on experiments demonstrated that the accumulation of TF mRNA requires de novo transcription of the TF gene . Since cycloheximide alone also caused an increase of TF mRNA levels and gene transcription it is concluded that the transcriptional activation of the TF gene does not require protein synthesis . Using specific protein kinase inhibitors, it was further demonstrated that activation of the protein kinase C pathway is involved in the induction of TF mRNA in HPBM . The accumulation of TF mRNA in LPS-stimulated HPBM is followed by an increase of TF activity on the cell surface . The kinetics of TF mRNA induction were found to be very similar in HPBM stimulated with LPS or PMA . However, in the latter case TF activity appeared considerably later on the cell membrane than in the LPS-stimulated cells . Non-stimulated HPBM contain very low levels of mRNA of the tissue factor pathway inhibitor (TFPI) . No induction of TFPI (mRNA, activity or antigen) in HPBM after LPS or PMA treatment was demonstrated . This seems to be in contrast with the earlier observation that the human monocyte cell line U937 produces significant amounts of TFPI in response to treatment with LPS and PMA. Int J Pept Protein Res, 1994 Apr, 43(4), 384 - 92 A free-energy simulation study of a bacterial collagenase inhibitor; Gilquin B et al.; The cis-trans conformational isomerisation of the N-methylated peptide bond of a bacterial collagenases peptide inhibitor (HS-CH2-CH2-CO-Pro-NMe-Ala) has been investigated by molecular dynamics simulations, energy minimisations and free-energy simulations in presence of solvent molecules . The free-energy difference between the cis and trans forms obtained by the thermodynamics integration method is equal to 0.95 kcal/mol in favour of the trans form, in accord with the experimental result . The main contribution to this free-energy difference comes from solute-water electrostatic interactions . Interestingly, we show that the number of interactions between water molecules and the oxygen atoms of the inhibitor is larger in the trans form than in the cis . Thus the organisation of water molecules around the inhibitor appears crucial in determining the population of the cis and trans conformers. Minerva Med, 1994 Apr, 85(4), 193 - 5 {Bacterial endocarditis due to Erysipelothrix rhusiopathiae . A clinical case report}; Iovinella V et al.; The authors describe an unusual case of bacterial endocarditis . A very rare etiologic agent, Erysipelothrix rhusiopathiae, was isolated . The patient was not fisherman or butcher and he did ot other risky job . Moreover the disease had an atypical course. Rinsho Shinkeigaku, 1994 Apr, 34(4), 331 - 5 {The significance of elevated Mn SOD level in cerebrospinal fluid of patients with bacterial meningitis--its relation to cytokine}; Hirose Y et al.; We measured cerebrospinal fluid (CSF) levels of manganese superoxide dismutase (Mn SOD) using an enzyme immunoassay method in 19 patients with bacterial meningitis (BM), 33 with aseptic meningitis (AM) and 13 with encephalitis (EN), and examined the significance of their elevations, especially in BM . 1) In BM, the Mn SOD levels were obviously high, ranging from 10.4 to 1179.2 ng/ml . The mean level of Mn SOD was 234.6 +/- 306.7 (SD) ng/ml and 18 patients showed abnormal levels of Mn SOD (more than 13.1 ng/ml) . On the other hand, in the remaining 2 diseases, the elevation of SOD levels was not remarkable: the mean levels of Mn SOD in AM and EN were 20.6 +/- 11.6 ng/ml and 41.9 +/- 23.6 ng/ml, respectively . 2) In AM and EN, Mn SOD levels well correlated with NSE or S-100b levels which are the markers of nervous tissue damages . But there was no correlation between the Mn SOD levels in BM and these markers . 3) In BM, there was a positive relationship between Mn SOD and total protein levels, but the disease days showing peak levels were different between them . In addition, Mn SOD levels showed no correlation with cell counts in CSF . 4) In BM, CSF levels of TNF-alpha and IL-1 alpha were remarkably high, whereas in AM and EN, the increases of these cytokines were not marked . And these cytokines in BM showed the peak values in the disease day before or when Mn SOD reached the peak levels.(ABSTRACT TRUNCATED AT 250 WORDS) Biotechniques, 1994 Apr, 16(4), 730 - 5 Immunoaffinity purification of FLAG epitope-tagged bacterial alkaline phosphatase using a novel monoclonal antibody and peptide elution; Brizzard BL et al.; The FLAG epitope is an eight amino acid peptide (AspTyrLysAspAspAspAspLys) that is useful for immunoaffinity purification of fusion proteins . A monoclonal antibody (anti-FLAG M1) that binds the FLAG epitope in a calcium-dependent manner and requires an N-terminal FLAG sequence has been described previously . We describe the use of a second anti-FLAG monoclonal antibody (anti-FLAG M2) in immunoaffinity purification of N-terminal Met-FLAG and C-terminal FLAG fusion to bacterial alkaline phosphatase . Although binding of an anti-FLAG M2 monoclonal antibody to the FLAG epitope is not calcium-dependent, bound fusion proteins can be eluted by competition with FLAG peptide. Plant Physiol, 1994 Apr, 104(4), 1411 - 7 Plant delta-aminolevulinic acid dehydratase . Expression in soybean root nodules and evidence for a bacterial lineage of the Alad gene; Kaczor CM et al.; We isolated a soybean (Glycine max) cDNA encoding the heme and chlorophyll synthesis enzyme delta-aminolevulinic acid (ALA) dehydratase by functional complementation of an Escherichia coli hemB mutant, and we designated the gene Alad . ALA dehydratase was strongly expressed in nodules but not in uninfected roots, although Alad mRNA was only 2- to 3-fold greater in the symbiotic tissue . Light was not essential for expression of Alad in leaves of dark-grown etiolated plantlets as discerned by mRNA, protein, and enzyme activity levels; hence, its expression in subterranean nodules was not unique in that regard . The data show that soybean can metabolize the ALA it synthesizes in nodules, which argues in favor of tetrapyrrole formation by the plant host in that organ . Molecular phylogenetic analysis of ALA dehydratases from 11 organisms indicated that plant and bacterial enzymes have a common lineage not shared by animals and yeast . We suggest that plant ALA dehydratase is descended from the bacterial endosymbiont ancestor of chloroplasts and that the Alad gene was transferred to the nucleus during plant evolution. Can J Vet Res, 1994 Apr, 58(2), 138 - 43 Comparison of the effects of ketoprofen and flunixin meglumine on the in vitro response of equine peripheral blood monocytes to bacterial endotoxin; Jackman BR et al.; The purpose of this study was to investigate the in vitro effects of flunixin meglumine, a cyclo-oxygenase inhibitor, and ketoprofen, a reported cyclo-oxygenase and lipoxygenase inhibitor, on the synthesis of cyclo-oxygenase end-products thromboxane B2 and prostaglandin E2, lipoxygenase derived 12-hydroxyeicosatetraenoic acid, tumor necrosis factor and tissue factor . Six adult horses were each randomly administered flunixin meglumine (1.1 mg/kg) or ketoprofen (2.2 mg/kg) intravenously every 12 hours with the drug treatments separated by two weeks . Blood samples were obtained prior to initiating treatment, the last day of treatment and for two consecutive days after the termination of treatment for measurement of serum concentrations of thromboxane B2 as well as isolation of peripheral blood monocytes . Quantitation of unstimulated, endotoxin- and calcium ionophore-induced synthesis of thromboxane B2, prostaglandin E2, 12-hydroxyeicosatetraenoic acid, tumor necrosis factor and tissue factor by peripheral blood monocytes was performed in vitro . Both flunixin meglumine and ketoprofen significantly decreased serum concentrations of thromboxane B2 demonstrating in vivo cyclo-oxygenase inhibition . There were no significant differences between drug treatment groups in the in vitro production of thromboxane B2, prostaglandin E2, 12-hydroxy-eicosatetraenoic acid, tumor necrosis factor or tissue factor . This study does not identify significant differences between the effects of flunixin meglumine and ketoprofen. Avian Dis, 1994 Apr-Jun, 38(2), 231 - 9 Bacterial colonization and in vivo expression of F1 (type 1) fimbrial antigens in chickens experimentally infected with pathogenic Escherichia coli; Dozois CM et al.; Escherichia coli strains that cause septicemia of poultry often possess F1 (type 1) fimbriae (encoded by pil {fim} homologous gene clusters) and/or P fimbriae (encoded by pap homologous gene clusters) . These fimbriae are thought to be involved in infection and colonization . To study the dynamics of infection due to E . coli with different virulence determinant profiles and to examine the expression of these fimbriae in vivo, three pathogenic E . coli isolates--O1 (pil+/pap+), O2 (pil+/pap), and O78 (pil+/pap+)--were administered intratracheally to 1.5-week-old chickens . Chickens were euthanatized from 3 to 144 hr after infection . The three isolates caused lesions in 30 to 55% of birds . Colonization rates of the trachea, lungs, internal organs, and pericardial fluid were similar for all three isolates, whereas significant differences among isolates were observed in colonization of the air sacs and blood . Bacteria appeared rapidly in the blood, liver, and spleen, whereas presence in the pericardial fluid generally occurred only after 24 hr postinoculation . The dynamics of colonization of the air sacs varied among isolates . Immunofluorescence of frozen tissue sections demonstrated F1 fimbriae (pil expressed) but not P fimbriae on all three isolates colonizing the trachea and on the O1 and O78 isolates colonizing the air sacs . Results suggest that F1 fimbriae are involved in the early stages of development of colisepticemia by promoting association of pathogenic E . coli with the trachea and air sacs of chickens. Actas Urol Esp, 1994 Apr, 18(4), 308 - 11 {Acute focal bacterial nephritis . Pathology mimicking renal masses}; Ruiz Dominguez JL et al.; Focal acute bacterial nephritis is an uncommon form of renal acute infection with a clinical presentation that, although similar to other acute infections of the renal parenchyma, must be taken into account in order to reach a correct differential diagnosis with other processes which have very similar radiological images but highly different management approaches, such as the renal carcinoma . This paper present one case in which the sings and symptoms were crucial to establish the diagnosis. J Hosp Infect, 1994 Apr, 26(4), 293 - 6 Dispensing surgical gloves onto the open surgical gown pack does not increase the bacterial contamination rate; Kong KC et al.; In implant surgery air and surface contamination have become important factors in post-operative wound infection . We established the rate of contamination of surgical gown packs and found that dropping gloves onto the open gown pack prior to scrubbing had no effect on it . Ninety-six contact plates were used for this study, which was carried out during clean orthopaedic operations in one operating room . The overall rate of contamination was 65% . Most of the contaminants were skin commensals . The rate of contamination was 67% when gloves were dropped onto the gown pack compared with 63% when opened and presented to the scrubbed and gowned theatre staff . However this difference was statistically not significant . The high rate of contamination was probably due to the gown packs having been left opened for too long . This delay arose because each gown pack had three gowns and would have been avoided if single gown packs had been used. Anal Biochem, 1994 Apr, 218(1), 204 - 9 Determination of free D-amino acids with a bacterial transaminase: their depletion leads to inhibition of bacterial growth; Jones WM et al.; A general procedure is described to determine the common free D-amino acids except D-proline in mixtures that also contain L-amino acids . The system employs exogenous pure bacterial D-amino acid transaminase coupled with 2-oxohexanoate, which accepts the amino group from D-amino acids to form D-norleucine . This amino acid is readily quantified by amino acid analysis since it elutes in a position not occupied by any of the common amino acids . Formation of norleucine denotes the presence of some D-amino acid(s) whose identity can be established by a corresponding decrease in the susceptible amino acid(s) after treatment . The utility of the procedure is demonstrated by determination of the amounts of free D-alanine and free D-glutamate in extracts of Escherichia coli JM-103 grown on minimal medium; D-alanine was the major D-amino acid . By the same principle, 2-oxohexanoate through coupling with endogenous bacterial D-amino acid transaminase is shown to be capable of inhibiting the growth of E . coli by depleting it of the D-alanine and D-glutamate. Mater Med Pol, 1994 Apr-Jun, 26(2), 49 - 53 Cellular and humoral components of neutrophil chemotaxis in neonates with bacterial infection; Chabior M; The initial stage of infection is characterized by both, changes in leucocyte function and humoral factors activity . In this paper we evaluated random migration and chemotaxis of neutrophils and serum ability to generate humoral factors influencing the neutrophil function . The studies were performed in 51 neonates during the acute phase of bacterial infection and in the remission time . Both random migration and chemotaxis were decreased during the acute phase of disease and normalized during remission . The same was true for serum ability to generate chemotactic activity . The serum of sick neonates did not alter the mobility of neutrophils isolated from adult healthy donors and it did not influence the normal chemotactic activity . We conclude, that the handicapped locomotive abilities of neutrophils in bacterial infection are temporary, and may require in severely ill infants supportive therapy to maintain adequate cellular and humoral immunity. J Clin Invest, 1994 Apr, 93(4), 1583 - 91 Bacterial lipopolysaccharide primes human neutrophils for enhanced release of arachidonic acid and causes phosphorylation of an 85-kD cytosolic phospholipase A2; Doerfler ME et al.; Production of leukotriene B4 (LTB4) by human neutrophils (PMN) in response to different stimuli is increased after pretreatment with lipopolysaccharides (LPS) . We have analyzed the steps in arachidonic acid (AA) metabolism affected by LPS by examining release of AA and its metabolites from {3H}AA prelabeled PMN . Pretreatment of PMN for 60 min with up to 1 microgram/ml of LPS alone had no effect, but release of {3H}AA was stimulated up to fivefold during subsequent stimulation with a second agent . In the absence of LPS-binding protein (LBP), priming was maximal after pretreatment of PMN with 10 ng of LPS/ml for 60 min; in the presence of LBP maximal priming occurred within 45 min at 0.1 ng of LPS/ml and within 15 min at 100 ng of LPS/ml . Treatment of PMN with 10 ng of LPS/ml also increased uptake of opsonized zymosan by up to 60% . Phospholipids are the source of released {3H}AA . No release was observed from {14C}oleic acid (OA)-labeled PMN suggesting that phospholipolysis may be specific for {3H}AA-labeled phospholipid pools . Cytosol from PMN primed with LPS contains two to three times the phospholipase A2 (PLA2) activity of control PMN, against 1-palmitoyl-{2-14C}arachidonoyl-phosphatidylcholine . This activity is Ca2+ dependent and dithiothreitol resistant . LPS priming is accompanied by reduced migration during SDS-PAGE of an 85-kD protein, identified as a cytosolic PLA2 . The extent and kinetics of this effect of LPS on cPLA2 parallel the priming of {3H}AA release, both depending on LPS concentration either with or without LBP . These findings suggest that priming by LPS of AA metabolism by PMN includes phosphorylation of an AA-phospholipid-selective cytosolic PLA2 that is dissociated from activation until a second stimulus is applied. J Leukoc Biol, 1994 Apr, 55(4), 483 - 8 Serum components enhance bacterial lipopolysaccharide-induced tissue factor expression and tumor necrosis factor-alpha secretion by bovine alveolar macrophages in vitro; Yang Z et al.; We have compared the effect of bacterial lipopolysaccharide (LPS) in combination with normal adult bovine serum (NBS), fetal bovine serum (FBS), or a bovine serum fraction on tissue factor expression and tumor necrosis factor alpha (TNF-alpha) secretion by bovine alveolar macrophages . At a concentration of 1 ng/ml, bacterial LPS alone failed to induce measurable tissue factor expression by the macrophages, but the presence of FBS, NBS, or a fraction of normal pooled bovine serum isolated by ion-exchange chromatography (fraction 2) markedly potentiated the effect of LPS . A protein concentration of 64 micrograms/ml NBS, 192 micrograms/ml FBS, and only 640 ng/ml fraction 2 was required to induce maximal tissue factor expression on the macrophages in combination with 1 ng/ml LPS . Comparison of quantities of added serum protein required to induce maximal potentiating effects indicated that fraction 2 was 100 times more potent than whole NBS and 300 times more potent than whole FBS . We similarly found that TNF-alpha secretion by macrophages exposed to LPS was responsive to serum and was highly responsive to fraction 2 . LPS alone (1 ng/ml) induced a relatively low level of TNF-alpha secretion by the macrophages, and the presence of FBS, NBS, or fraction 2 potentiated the effect of LPS . A concentration of 64.0 micrograms/ml NBS, 320.0 micrograms/ml FBS, and 3.2 micrograms/ml fraction 2 serum protein induced near-maximal TNF-alpha secretion by the macrophages . Comparison of the concentration of serum protein required to induce these potentiating effects indicated that fraction 2 was approximately 20 times more potent than whole NBS and 100 times more potent than whole FBS . The stimulatory effect of LPS plus fraction 2 serum proteins was dependent on the CD14 receptor, as monoclonal antibodies directed against CD14 (My4, 60bd; 10 micrograms/ml) inhibited tissue factor expression and TNF-alpha secretion by the macrophages. Proc Natl Acad Sci U S A, 1994 Mar 29, 91(7), 2448 - 55 Human ecology and behavior and sexually transmitted bacterial infections; Holmes KK; The three direct determinants of the rate of spread of sexually transmitted diseases (STDs) are sexual behaviors, the mean duration of infectiousness, and the mean efficiency of sexual transmission of each STD . Underlying ecological and behavioral factors that operate through one or more of these direct determinants lie on a continuum, ranging from those most proximate back to those more remote (in time or mechanism) from the direct determinants . Most remote and least modifiable are the historical stages of economic development that even today conspicuously influence patterns of sexual behavior . Next are the distribution and changing patterns of climate, hygiene, and population density; the global population explosion and stages of the demographic transition; and ongoing changes in human physiology (e.g., menarche at younger age) and culture (e.g., later marriage) . More proximate on the continuum are war, migration, and travel; and current policies for economic development and social welfare . Most recent or modifiable are technologic and commercial product development (e.g., oral contraceptives); circumcision, condom, spermicide, and contraception practices; patterns of illicit drug use that influence sexual behaviors; and the accessibility, quality, and use of STD health care . These underlying factors help explain why the curable bacterial STDs are epidemic in developing countries and why the United States is the only industrialized country that has failed to control bacterial STDs during the AIDS era. Proc Natl Acad Sci U S A, 1994 Mar 29, 91(7),