J Med Entomol, 1993 Nov, 30(6), 1050 - 2 Bacterial enrichment in the surface microlayer of an Anopheles quadrimaculatus (diptera: culicidae) larval habitat; Walker ED et al.; Sampling of the surface microlayer of water in a marsh habitat of Anopheles quadrimaculatus Say in Michigan revealed it to be enriched with bacteria compared with subsurface water samples . Concentrations of total numbers of bacteria ranged from 18.8 x 10(6)/ml to 65.4 x 10(6)/ml (mean, 35.9 x 10(6)/ml; n = 13) in surface microlayer samples, and from 3.8 x 10(6)/ml to 14.3 x 10(6)/ml, (mean, 7.8 x 10(6)/ml; n = 13) in subsurface samples . Specifically, the surface microlayer had higher concentrations of three morphotypes of bacteria (cocci, rods, and rods attached to detritus particles) . Given that bacteria are an important food of mosquito larvae, we suggest that the interfacial feeding behavior of An . quadrimaculatus larvae allows them to exploit the surface microlayer, a food-rich zone. Kansenshogaku Zasshi, 1993 Nov, 67(11), 1083 - 93 {Studies of neutrophil function in the elderly: especially analysis of bedridden patients and patients with bacterial infection}; Adachi S et al.; To evaluate the host defense function in the elderly, chemotaxis and chemiluminescence (CL) were assessed as indicators of neutrophil function in healthy and bedridden subjects as well as in patients with bacterial infection . In addition, humoral host defensive factors were also investigated . 1) The differences in chemotaxis and CL between healthy elderly subjects and healthy adults were slight . Neutrophil function showed little decrease with aging . 2) Plasma fibronectin (PFN) values in bedridden patients were lower than in healthy elderly subjects . 3) Chemotaxis and CL in patients in the acute phase of bacterial infection were higher than those of healthy elderly subjects regardless of whether these patients were bedridden or not . CL was slightly lower in bedridden patients than in non-bedridden patients . CL and body temperature were positively correlated in the acute phase of bacterial infection . Among the humoral defensive factors, PFN revealed lowered values in the acute phase of bacterial infection . 4) As the infection healed, chemotaxis and CL decreased, while PFN rose to normal . In the complement system, CH50 increased slightly as the infection healed . From these results, we concluded that neutrophil function in elderly subjects, regardless of whether they were bedridden or not, is enhanced by infection . Responses to infection of bedridden patients were slightly weaker than those of non-bedridden subjects. J Antibiot (Tokyo), 1993 Nov, 46(11), 1764 - 6 An assay for the detection of bacterial DNA gyrase inhibitors; Osburne MS et al.; In summary, we have developed a sensitive detection system for inhibitors of bacterial DNA gyrase . The use of B . subtilis as the host organism confers the advantage that it is sensitive to both gyrase subunit A and B inhibitors, whereas E . coli is relatively insensitive to B subunit inhibitors in vivo . Using this assay, we identified a new DNA gyrase inhibitor with a novel structure. FEMS Microbiol Rev, 1993 Nov, 12(4), 293 - 314 Bacterial solute transport proteins in their lipid environment; Veld GI et al.; The cytoplasmic membrane of bacteria is a selective barrier that restricts entry and exit of solutes . Transport of solutes across this membrane is catalyzed by specific membrane proteins . Integral membrane proteins usually require specific lipids for optimal activity and are inhibited by other lipid species . Their activities are also sensitive to the lipid bilayer dynamics and physico-chemical state . Bacteria can adapt to changes in the environments (respective temperature, hydrostatic pressure, and pH) by altering the lipid composition of the membrane . Homeoviscous adaptation results in the maintenance of the liquid-crystalline phase through alterations in the degree of acyl chain saturation and branching, acyl chain length and the sterol content of the membrane . Homeophasic adaptation prevents the formation of non-bilayer phases, which would disrupt membrane organization and increase permeability . A balance is maintained between the lamellar phase, preferring lipids, and those that adopt a non-bilayer organization . As a result, the membrane proteins are optimally active under physiological conditions . The molecular basis of lipid-protein interactions is still obscure . Annular lipids stabilize integral membrane proteins . Stabilization occurs through electrostatic and possibly other interactions between the lipid headgroups and the charged amino acid residues close to the phospholipid-water interface, and hydrophobic interactions between the fatty acyl chains and the membrane-spanning segments . Reconstitution techniques allow manipulation of the lipid composition of the membrane in a way that is difficult to achieve in vivo . The physical characteristics of membrane lipids that affect protein-mediated transport functions have been studied in liposomal systems that separate an inner and outer compartment . The activity of most transport proteins is modulated by the bulk physical characteristics of the lipid bilayer, while specific lipid requirements appear rare. Plant Mol Biol, 1993 Nov, 23(4), 759 - 68 Concerted regulation of lysine and threonine synthesis in tobacco plants expressing bacterial feedback-insensitive aspartate kinase and dihydrodipicolinate synthase; Shaul O et al.; The essential amino acids lysine and threonine are synthesized in higher plants by two separate branches of a common pathway . This pathway is primarily regulated by three key enzymes, namely aspartate kinase (AK), dihydrodipicolinate synthase (DHPS) and homoserine dehydrogenase (HSD), but how these enzymes operate in concert is as yet unknown . Addressing this issue, we have expressed in transgenic tobacco plants high levels of bacterial AK and DHPS, which are much less sensitive to feedback inhibition by lysine and threonine than their plant counterparts . Such expression of the bacterial DHPS by itself resulted in a substantial overproduction of lysine, whereas plants expressing only the bacterial AK overproduced threonine . When both bacterial enzymes were expressed in the same plant, the level of free lysine exceeded by far the level obtained by the bacterial DHPS alone . This increase, however, was accompanied by a significant reduction in threonine accumulation compared to plants expressing the bacterial AK alone . Our results suggested that in tobacco plants the synthesis of both lysine and threonine is under a concerted regulation exerted by AK, DHPS, and possibly also by HSD . We propose that the balance between lysine and threonine synthesis is determined by competition between DHPS and HSD on limiting amounts of their common substrate 3-aspartic semialdehyde, whose level, in turn, is determined primarily by the activity of AK . The potential of this molecular approach to increase the nutritional quality of plants is discussed. Physiol Behav, 1993 Nov, 54(5), 961 - 6 Does a learned taste aversion contribute to the anorectic effect of bacterial lipopolysaccharide? Weingarten S, Senn M, Langhans W. The present study addressed the possible role of a conditioned taste aversion in the anorectic effect of bacterial lipopolysaccharide (LPS) in the rat . Pairing an intraperitoneal (IP) injection of LPS (100 micrograms/kg b.wt.) with the subsequent presentation of a familiar diet (FD) or of a novel-tasting saccharin diet (SD) for several hours did not affect FD or SD intake when the same diet was offered several days later after 12 h of food deprivation . However, food intake during the second presentation of SD was reduced when food was not withheld prior to the test . In a similarly designed experiment, the antipyretic and antiinflammatory drug indomethacin (5 mg/kg b.wt., IP) attenuated the anorectic effect of LPS during the initial pairing, but did not affect the inhibition of SD intake in LPS-pretreated rats during the second feeding test . The antiemetic trimethobenzamide (5 mg/kg b.wt., IP) failed to influence the anorectic effect of LPS . Lesion of the area postrema (AP) and the adjacent nucleus of the solitary tract (NST) was found to enhance the anorectic effect of LPS, but the development of tolerance to this effect remained unchanged in AP/NST-lesioned animals . In spite of the ability of LPS to induce a taste aversion that inhibits feeding under certain conditions (novel-tasting diet, no food deprivation prior to the feeding test), the findings indicate that a learned taste aversion is not the only contributor to the anorectic effect of LPS. Oral Surg Oral Med Oral Pathol, 1993 Nov, 76(5), 661 - 3 Bacterial contamination of dental radiographic film; Bajuscak RE et al.; Three types of paper-covered and one type of plastic-covered Kodak dental film were used to determine if bacteria could penetrate the coverings and contaminate the inner film . Films from each group were immersed for 30 or 120 seconds in high concentrations of known bacterial suspensions with or without 10% sterilized calf serum added to the incubation media . The plastic-covered film effectively excluded all bacteria, whereas the paper-covered film showed contamination with all organisms even at 30 seconds . Increasing the viscosity of the incubation medium with calf serum decreased the level of contamination. Am J Physiol, 1993 Nov, 265(5 Pt 2), R1179 - 83 Fever and thermogenesis in response to bacterial endotoxin involve macrophage-dependent mechanisms in rats; Derijk RH et al.; Increases in thermogenesis and body temperature (fever) frequently accompany infection or injury and are thought to be mediated by endogenous pyrogens (e.g . cytokines), which are released from activated immune cells such as macrophages . Therefore, we have investigated the effect of selective elimination of peripheral macrophages on the changes in oxygen consumption (VO2) and colonic temperature in response to bacterial lipopolysaccharide (LPS) in the rat . Peripheral macrophages were depleted by intravenous injection of liposomes containing the drug dichloromethylene diphosphonate (Cl2MDP) . Resting oxygen consumption and colonic temperatures were not affected by macrophage elimination . In intact rats, peripheral injection of LPS (0.1-0.5 mg/kg) elicited an increase in colonic temperature and in oxygen consumption that declined at higher doses (2.5 mg/kg) . The pyrogenic and thermogenic responses to LPS were significantly attenuated in rats in which peripheral macrophages were eliminated . Previously, we have reported that elimination of macrophages blunts the plasma interleukin-1 (IL-1) response to LPS . Here we show that elimination of macrophages does not affect the increase in plasma IL-6 concentrations in response to LPS . These data indicate that the pyrogenic and thermogenic responses to LPS are at least in part dependent on mechanisms involving peripheral macrophages, and that peripherally produced IL-1 rather than IL-6 may be an important mediator of the changes in oxygen consumption and colonic temperature in response to LPS. Am J Obstet Gynecol, 1993 Nov, 169(5), 1161 - 6 Endotoxin and interleukin-1 alpha in the cervical mucus and vaginal fluid of pregnant women with bacterial vaginosis; Platz-Christensen JJ et al.; OBJECTIVE: The purpose of our study was to determine the concentrations of endotoxin and interleukin-1 alpha in the cervical mucus and vaginal fluid of pregnant women who either did or did not have bacterial vaginosis . STUDY DESIGN: Samples of cervical mucus and vaginal fluid were collected from women in early pregnancy who had signs of bacterial vaginosis and from healthy control subjects . The samples were analyzed for the concentrations of endotoxin and interleukin-1 alpha . In addition, wet mounts were examined for signs of inflammation indicated by increased numbers of leukocytes . RESULTS: Both endotoxin and interleukin-1 alpha occurred in much higher concentrations (p < 0.0001, p < 0.0002) in both the cervical mucus and the vaginal fluid of women with signs of bacterial vaginosis than they did in healthy control subjects . A correlation was found between the interleukin-1 alpha concentrations in the vaginal fluid and the number of leukocytes as judged by a semi-quantitative evaluation of wet mounts (p = 0.0365) . The concentrations of endotoxin correlated with those of interleukin-1 alpha in both fluids (vaginal fluid, p < 0.01; cervical mucus, p < 0.01) . CONCLUSION: Our study shows that concentrations of endotoxin and interleukin-1 alpha in cervical mucus and vaginal fluid of women in early pregnancy who have bacterial vaginosis are significantly higher than the corresponding levels in control subjects. Am J Obstet Gynecol, 1993 Nov, 169(5), 1130 - 4 Coitus during pregnancy is not related to bacterial vaginosis or preterm birth; Kurki T et al.; OBJECTIVE: Our goal was to assess the relationship between bacterial vaginosis and sexual intercourse and the impact of both on preterm birth . STUDY DESIGN: The presence of bacterial vaginosis was assessed in 790 healthy nulliparous women between 8 and 17 weeks' gestation, and they were then asked to record weekly the number of occasions of sexual intercourse from registration to term . The end point for each was the occurrence of preterm uterine contractions, preterm birth, or preterm rupture of the membranes . RESULTS: A total of 407 women returned the specific follow-up charts on vaginal intercourse . Bacterial vaginosis was detected at registration in 93 of the 407 subjects (22.9%), but the presence or absence of bacterial vaginosis had no predictive value as regards the frequency of subsequent intercourse . Of the 39 women (9.6%) who experienced threatened preterm labor, 16 were delivered before 37 weeks of gestation . In addition, premature rupture of membranes occurred in 48 women (9 preterm and 39 term) . The 95% confidence interval for the weekly frequency of intercourse was calculated for those women delivered at term; a larger (p < 0.05) proportion of women delivered preterm, with (6/9, 66.7%) or without (13/16, 81.3%) premature rupture of membranes, had frequencies less than the 95% confidence interval during the second trimester of pregnancy; the respective proportions were 75.0% (6/8) and 78.6% (11/14) during the third trimester . Furthermore, among those women who had stopped having sexual intercourse in the last trimester, a larger (p < 0.05) proportion was delivered preterm (57.1%), with or without premature rupture of membranes, than was delivered at term (13.6%) . In addition, only three women (18.8%) delivered preterm, with (two women) or without (one woman) premature rupture of membranes, reported having intercourse within 1 week before the onset of complications . CONCLUSION: For healthy nulliparous women, coitus during pregnancy is not related to bacterial vaginosis and does not predispose to preterm birth. Proc Natl Acad Sci U S A, 1993 Nov 1, 90(21), 9892 - 5 Occurrence of fragmented 16S rRNA in an obligate bacterial endosymbiont of Paramecium caudatum; Springer N et al.; The phylogenetic position of Caedibacter caryophila, a so far noncultured killer symbiont of Paramecium caudatum, was elucidated by comparative sequence analysis of in vitro amplified 16S rRNA genes (rDNA) . C . caryophila is a member of the alpha subclass of the Proteobacteria phylum . Within this subclass C . caryophila is moderately related to Holospora obtusa, which is another obligate endosymbiont of Paramecium caudatum, and to Rickettsia . A 16S rRNA targeted specific hybridization probe was designed and used for in situ detection of C . caryophila within its host cell . Comparison of the 16S rDNA primary structure of C . caryophila with homologous sequences from other bacteria revealed an unusual insertion of 194 base pairs within the 5'-terminal part of the corresponding gene . The intervening sequence is not present in mature 16S rRNA of C . caryophila . It was demonstrated that C . caryophila contained fragmented 16S rRNA. J Pediatr, 1993 Nov, 123(5), 801 - 10 Bacterial polysaccharide immune globulin for prophylaxis of acute otitis media in high-risk children; Shurin PA et al.; We evaluated the prevention of recurrences of acute otitis media (AOM) by bacterial polysaccharide immune globulin (BPIG), a hyperimmune human immune globulin prepared by immunizing donors with bacterial polysaccharide vaccines . We used a randomized, stratified, double-blind, placebo-controlled design . Children < or = 24 months of age with 1 to 3 prior episodes of AOM received BPIG, 0.5 ml/kg, or saline placebo intramuscularly at entry and 30 days later . During the 120-day follow-up period, AOM was diagnosed by using clinical criteria and was confirmed with tympanocentesis and culture of the middle ear exudates . Eighty-eight episodes of AOM were observed in 76 patients who completed the study . The incidence of AOM during the entire 120-day study period was similar in BPIG and placebo recipients . Pneumococcal AOM was significantly less frequent in BPIG recipients (0.21 episode per patient) than in placebo recipients (0.45 episode per patient; p = 0.05) . Time spent free of AOM was significantly prolonged in recipients of BPIG, in comparison with placebo recipients (51 vs 35 days; p = 0.034) . This study demonstrated that circulating antibody, even without stimulation of specific local immunity, may prevent infection of the middle ear . The use of immune globulin preparations for longer periods or at a higher dosage might decrease the incidence of recurrent AOM in otitis-prone children, and deserves further evaluation. J Nucl Med, 1993 Nov, 34(11), 1975 - 9 Localization of indium-111-immunoglobulin G, technetium-99m-immunoglobulin G and indium-111-labeled white blood cells at sites of acute bacterial infection in rabbits; Barrow SA et al.; Biodistribution and infection imaging properties of 111In-DTPA-IgG, 99mTc-hydrazino nicotinamide-IgG and 111In-WBC were compared in rabbits with E . coli infection . Groups of six rabbits were injected with 10 mCi of 99mTc-IgG plus 0.5 mCi of 111In-IgG or 1 mCi of 99mTc-IgG plus 0.05 mCi of 111In-WBC . At 4-5 and 18-20 hr, dual photon scintigrams were acquired . At both times, the distributions of 99mTc and 111In-IgG were nearly identical . The sites of infection were well visualized with all three radiopharmaceuticals . In the early images, the target-to-background ratios (T/B) for 111In and 99mTc-IgG determined by ROI analysis were 1.95 +/- 0.26 and 2.57 +/- 0.38 (p = NS) . In the delayed images, the T/B ratios increased (p < 0.01) to 3.56 +/- 0.49 and 4.90 +/- 0.98 . At both times, the T/B ratios for 111In-WBC were higher (p < 0.01); 4.17 +/- 0.78 at 4-5 hr and 8.52 +/- 1.52 at 18-20 hr . These results indicate that all three agents yield excellent images of infection sites . Although 111In-WBC had higher T/B ratios, the ease of preparation of the radiolabeled proteins makes them attractive alternatives for infection imaging. J Nucl Med, 1993 Nov, 34(11), 1964 - 74 Technetium-99m-labeled hydrazino nicotinamide derivatized chemotactic peptide analogs for imaging focal sites of bacterial infection; Babich JW et al.; We synthesized and evaluated four hydrazino nicotinamide (HYNIC) derivatized chemotactic peptide analogs: For-NleLFK-HYNIC (HP1), For-MLFK-HYNIC (HP2), For-MLFNH(CH2)6NH-HYNIC (HP3), and For-MLF-(D)-K-HYNIC (HP4), for in vitro bioactivity and receptor binding . The peptides were radiolabeled with 99mTc via a glucoheptonate co-ligand and their biodistribution determined in rats (n = 6/time point) at 5, 30, 60 and 120 min after injection . Localization of the peptides at sites of deep thigh Escherichia coli infection was determined by radioactivity measurements on excised tissues in rats (n = 6/time point) and rabbits as well as scintillation camera imaging in rabbits (n = 6) . All peptides maintained biological activity (EC50s for O2 production by human PMNs: 12-500 nM) and the ability to bind to the oligopeptide chemoattractant receptor on human PMNs (EC50s for binding: 0.12-40 nM) . After incubation with 99mTc-glucoheptonate, radiolabeled peptides were isolated by HPLC at specific activities of > 10,000 mCi/microM . Technetium-99m-labeled peptides retained receptor binding with EC50s < 10 nM . Blood clearance of all four peptides was rapid . Biodistributions of the individual peptides were similar, with low levels of accumulation in most normal tissues . In rats, all of the peptides concentrated at the infection sites (T/B ratio: 2.5-3:1) within 1 hr of injection . In rabbits, outstanding images of the infection sites were obtained, with T/B ratios of > 20:1 at 15 hr after injection . This study demonstrates that 99mTc-labeled chemotactic peptide analogs are effective agents for the external imaging of focal sites of infection. Eur J Immunol, 1993 Nov, 23(11), 2998 - 3002 Induction of T cell responses by chimeric bacterial proteins expressing several copies of a viral T cell epitope; Lo-Man R et al.; A viral T cell epitope was genetically inserted within the periplasmic MalE protein of Escherichia coli in two different permissive insertion sites and resulting hybrid proteins were used to study the in vitro and in vivo immunogenicity of the foreign T cell epitope . Purified hybrid MalE proteins containing the T cell epitope 120-132 (PreS:T) from PreS2 region of hepatitis B virus HBsAg inserted alone or with its adjacent B cell epitope (132-145) were able to induce strong peptide-specific T cell responses in mice . In vitro stimulation of primed lymph node cells or specific T cell hybridomas by the hybrid proteins required processing of the inserted T cell epitope and was inhibited by antigen-presenting cells fixation . The inserted T cell epitope was presented in vitro, in association with appropriate major histocompatibility complex molecules, as efficiently as free synthetic peptide . The in vitro immunogenicity of MalE hybrid proteins was increased by inserting four tandemly repeated copies of PreS:T, either at site 133 or 303 . These results were confirmed in vivo by comparing the proliferative responses of lymph node cells from DBA/1 mice primed with MalE hybrid proteins containing one or four copies of PreS:T . Thus, the use of MalE hybrid proteins expressing multiple copies of a given foreign T cell epitope allows the induction of peptide-specific T cell response with a lower dose of priming antigen. Bull Acad Natl Med, 1993 Nov, 177(8), 1373 - 9; discussion 1379-80 {Importance of bacterial vaccines}; Meyran M; The vaccines presently developed in laboratories are more numerous than the ones in current use . This acceleration of research efforts was made possible by a genuine biotechnological revolution which opened new avenues for vaccine preparation, with techniques such as the selection of relevant antigens and the use of subunit vaccines . Genetic engineering plays an increasing part in the development and production of both live and inactivated vaccines. Trends Microbiol, 1993 Nov, 1(8), 306 - 10 The essential tension: opposed reactions in bacterial two-component regulatory systems; Russo FD et al.; In many different bacteria several sensory-response functions are controlled by systems of similar design . Most consist of two proteins, one of which regulates the phosphorylation of the other in response to an environmental stimulus . Regulation is achieved by balancing opposed phosphorylation and dephosphorylation reactions against each other . Remarkably, such a system can generate a signal whose strength is independent of the concentration of either component. Immunol Lett, 1993 Nov, 38(3), 173 - 7 Comparative analysis of serological activity of non-structural protein (NS1) from tick-borne encephalitis virus and its analog expressed in bacterial cells; Pressman EK et al.; By means of immunoaffinity chromatography and expression of the gene in Escherichia coli, non-structural glycoprotein NS1 of tick-borne encephalitis virus (TBEV) and its recombinant analog were prepared . Antisera against these proteins were obtained by hyperimmunisation of rabbits . The antisera were tested by means of complement fixation, agar diffusion, hemagglutination inhibition and virus neutralization . Although both antisera are reacted with natural antigen, the recombinant analog of NS1 did not bind antibodies against natural protein in complement fixation and immunoprecipitation . Nevertheless the NS1 analog was rather active in ELISA . Neither the natural nor the recombinant protein protected experimental animals from lethal virus infection . A contamination of natural NS1 antigen with small amounts of structural glycoprotein E may be responsible for both antibody formation and virus neutralization . This can be relevant for the design of a subunit vaccine. Ital J Gastroenterol, 1993 Nov-Dec, 25(9), 490 - 6 Orocaecal transit, bacterial overgrowth and hydrogen production in diabetes mellitus; Sarno S et al.; Orocaecal transit time was investigated using the hydrogen breath test in 39 insulin-requiring patients with long-standing Type I diabetes mellitus and 26 healthy control subjects . Thirty four patients complained of different gastrointestinal symptoms . The standard meal consisted of 10 g lactulose in 150 ml tap water . Mean transit time was significantly longer in the patient group (106.4 +/- 31.1 min) than in control subjects (84.2 +/- 27.1 min), and differences in OCTT between symptomatic subgroups were also significant . No correlation was found between orocaecal transit time and gastric emptying of a solid meal measured with scintigraphic method, HbA1c values, and other signs of automatic and peripheral neuropathy . The incidence of bacterial overgrowth among the diabetics was minimal . The percentage of H2 non-producers did not significantly differ between control and patient groups (23% and 26%, respectively) . The absolute amount of breathed hydrogen was, however, significantly lower in diabetics at all time intervals . This indicates that specific changes in hydrogen production may be related to pathophysiological features as a consequence or as an associated symptom. J Comp Pathol, 1993 Nov, 109(4), 429 - 32 A prolonged period of uninterrupted protein synthesis is essential for survival in mice given a submicrogram dose of bacterial endotoxin; Parry EW; A majority of mice failed to survive challenge with a normally tolerated dose of cycloheximide given at intervals up to 96 h after a single intraperitoneal injection of 0.02 microgram of S . enteritidis endotoxin . Only by 168 h did a majority of endotoxin-pretreated animals resist such a challenge . This finding underlines the essential role of uninterrupted protein synthesis in resistance to the lethal effects of endotoxin, and demonstrates the relatively persistent nature of this requirement after systemic exposure to an extremely low dose of lipopolysaccharide. Ir Med J, 1993 Nov-Dec, 86(6), 208 - 9 Moraxella catarrhalis--an unusual pathogen in bacterial tracheitis; Bodkin S et al.; We report a case of bacterial tracheitis caused by Moraxella Catarrhalis . We are unaware of any previous similar report from this country. J Bacteriol, 1993 Nov, 175(21), 7016 - 23 Mutational analysis of the bacterial signal-transducing protein kinase/phosphatase nitrogen regulator II (NRII or NtrB); Atkinson MR et al.; The signal-transducing kinase/phosphatase nitrogen regulator II (NRII or NtrB) is required for the efficient positive and negative regulation of glnA, encoding glutamine synthetase, and the Ntr regulon in response to the availability of ammonia . Alteration of highly conserved residues within the kinase/phosphatase domain of NRII revealed that the positive and negative regulatory functions of NRII could be genetically separated and that negative regulation by NRII did not require the highly conserved His-139, Glu-140, Asn-248, Asp-287, Gly-289, Gly-291, Gly-313, or Gly-315 residue . These mutations affected the positive regulatory function of NRII to various extents . Certain substitutions at codons 139 and 140 resulted in mutant NRII proteins that were transdominant negative regulators of glnA and the Ntr regulon even in the absence of nitrogen limitation . In addition, we examined three small deletions near the 3' end of the gene encoding NRII; these resulted in altered proteins that retained the negative regulatory function but were defective to various extents in the positive regulatory function . A truncated NRII protein missing the C-terminal 59 codons because of a nonsense mutation at codon 291 lacked entirely the positive regulatory function but was a negative regulator of glnA even in the absence of nitrogen limitation . Thus, we have identified both point and deletion mutations that convert NRII into a negative regulator of glnA and the Ntr regulon irrespective of the nitrogen status of the cell. Arch Oral Biol, 1993 Nov, 38(11), 931 - 5 Simultaneous hybridization and subsequent colour detection of subgingival bacterial DNA on colony lifts; Cross DL et al.; This paper reports the development of a protocol allowing hybridization and detection of DNA fixed to nylon colony lifts from up to three species of bacteria simultaneously . Half ml samples of serial dilutions of pure cooked-meat broth (CMB) cultures of Capnocytophaga ochracea, Actinobacillus actinomycetemcomitans and Prevotella intermedia were grown on trypticase soy blood agar (TSBA) plates for 7 days in an anaerobic chamber . From the same CMBs a further set of dilutions was completed that contained all three species . Samples from these dilutions produced mixed-growth TSBA plates following anaerobic incubation for 7 days . After incubation, colony counts on pure-growth TSBA plates were enumerated by colony counter . Colony counts of C . ochracea, A . actinomycetemcomitans and P . intermedia on mixed-growth TSBA plates were enumerated by nylon colony lift, simultaneous hybridization with non-isotopic whole chromosomal DNA probes and alkaline phosphatase substrates generating three colours . The results indicate that the protocol correctly identified and differentiated between the three species on mixed-growth TSBA plates . The proportions of each species and mean total colony count expected by counting pure plates were in agreement with the proportions of each species and total colony counts enumerated by DNA probes on mixed growth plates . The development of simultaneous hybridization and multicolour detection will result in improved data recovery from dental plaque samples, in addition to reducing the cost and labour required in current colony-lift protocols, without affecting the specificity or sensitivity of the probes used. Science, 1993 Oct 22, 262(5133), 566 - 9 A yeast protein similar to bacterial two-component regulators; Ota IM et al.; Many bacterial signaling pathways involve a two-component design . In these pathways, a sensor kinase, when activated by a signal, phosphorylates its own histidine, which then serves as a phosphoryl donor to an aspartate in a response regulator protein . The Sln1 protein of the yeast Saccharomyces cerevisiae has sequence similarities to both the histidine kinase and the response regulator proteins of bacteria . A missense mutation in SLN1 is lethal in the absence but not in the presence of the N-end rule pathway, a ubiquitin-dependent proteolytic system . The finding of SLN1 demonstrates that a mode of signal transduction similar to the bacterial two-component design operates in eukaryotes as well. Biochim Biophys Acta, 1993 Oct 19, 1216(1), 81 - 93 Molecular cloning and bacterial expression of cDNA for rat calpain II 80 kDa subunit; DeLuca CI et al.; The complete cDNA of 3.2 kb for rat calpain II large subunit has been constructed from library- and polymerase chain reaction-derived fragments, and sequenced . The cDNA encodes a protein of 700 amino acids having 93% sequence identity with human calpain II, and 61% identity with human calpain I . The gene possesses 21 exons, of which exons 3-21 have been mapped over 33 kb of the rat genome . A new phagemid expression vector was created from pT7-7 by insertion of the f1 origin and mutation of an NdeI to an NcoI site . Rat calpain II cDNA ligated into this vector expressed in Escherichia coli an 80 kDa protein identical in size to highly purified rat calpain II; this protein was specifically recognized on immunoblots by an affinity-purified anti-rat calpain II antibody . This is the second mammalian calpain II large subunit to be fully sequenced, and the first to be artificially expressed. Biochem Biophys Res Commun, 1993 Oct 15, 196(1), 167 - 72 Bacterial synthesis of truncated forms of the human vitamin D receptor and characterization of anti-receptor monoclonal antibodies; Schaefer-Klein J et al.; We biosynthesized full-length (amino acids 1-427) and truncated human 1,25-dihydroxyvitamin D3 receptor proteins that encompassed only the putative DNA binding domain (amino acids 1-112) or the DNA binding domain and parts of the sterol binding domain (amino acids 1-193 and 1-328) in a bacterial expression system . We also prepared monoclonal antibodies against the full-length vitamin D receptor . The binding properties of the monoclonal antibodies were characterized by their ability to bind to full-length and truncated vitamin D receptor protein constructs . Seven of twelve monoclonal antibodies recognized the full-length receptor protein . These antibodies bound to truncated hVDR proteins with decreasing affinities as successive truncations were made from the carboxy-terminal end of the receptor protein . The five remaining monoclonal antibodies recognized the full-length and truncated receptor proteins with equally low affinities . Truncated forms of the vitamin D receptor and region-specific antibodies will be useful in assessing the properties of the receptor. Br J Hosp Med, 1993 Oct 6-19, 50(7), 403 - 7 Current trends in the management of bacterial meningitis; Nathavitharana KA et al.; The relevance of the host inflammatory response in the pathophysiology of bacterial meningitis is an important new concept . Modulation of this process by steroids may significantly affect the prognosis . We examine current trends in the clinical management of bacterial meningitis. J Cell Physiol, 1993 Oct, 157(1), 13 - 23 Bacterial lipopolysaccharide induces actin reorganization, intercellular gap formation, and endothelial barrier dysfunction in pulmonary vascular endothelial cells: concurrent F-actin depolymerization and new actin synthesis; Goldblum SE et al.; Bacterial lipopolysaccharide (LPS) influences pulmonary vascular endothelial barrier function in vitro . We studied whether LPS regulates endothelial barrier function through actin reorganization . Postconfluent bovine pulmonary artery endothelial cell monolayers were exposed to Escherichia coli 0111:B4 LPS 10 ng/ml or media for up to 6 h and evaluated for: 1) transendothelial 14C-albumin flux, 2) F-actin organization with fluorescence microscopy, 3) F-actin quantitation by spectrofluorometry, and 4) monomeric G-actin levels by the DNAse 1 inhibition assay . LPS induced increments in 14C-albumin flux (P < 0.001) and intercellular gap formation at > or = 2-6 h . During this same time period the endothelial F-actin pool was not significantly changed compared to simultaneous media controls . Mean (+/- SE) G-actin (micrograms/mg total protein) was significantly (P < 0.002) increased compared to simultaneous media controls at 2, 4, and 6 h but not at 0.5 or 1 h . Prior F-actin stabilization with phallicidin protected against the LPS-induced increments in G-actin (P = 0.040) as well as changes in barrier function (P < 0.0001) . Prior protein synthesis inhibition unmasked an LPS-induced decrement in F-actin (P = 0.0044), blunted the G-actin increment (P = 0.010), and increased LPS-induced changes in endothelial barrier function (P < 0.0001) . Therefore, LPS induces pulmonary vascular endothelial F-actin depolymerization, intercellular gap formation, and barrier dysfunction . Over the same time period, LPS increased total actin (P < 0.0001) and new actin synthesis (P = 0.0063) which may be a compensatory endothelial cell response to LPS-induced F-actin depolymerization. Eur J Immunol, 1993 Oct, 23(10), 2700 - 3 Interleukin-10 inhibits the induction of monocyte procoagulant activity by bacterial lipopolysaccharide; Pradier O et al.; Monocytes stimulated with bacterial lipopolysaccharide (LPS) generate a procoagulant activity (PCA) related to the induction of tissue factor (TF) expression at their surface . Since interleukin-10 (IL-10) was recently shown to inhibit LPS-induced cytokine production and is currently considered as a potential therapeutic agent in septic shock, we were interested to determine its effects on LPS-induced monocyte PCA . Peripheral blood mononuclear cells (PBMC) from healthy donors were incubated with 1 microgram/ml LPS in the presence of serial dilutions of recombinant human IL-10 and PCA was determined after 6 h in a one-stage clotting assay . IL-10 inhibited in a dose-dependent manner LPS-induced TF-dependent PCA: a significant effect was already observed with 30 pg/ml IL-10 while 64-97% inhibition was achieved with 120 pg/ml IL-10 . In parallel flow cytometry experiments, IL-10 was shown to block LPS-induced TF expression at the surface of monocytes . In order to inhibit LPS-induced PCA, IL-10 had to be added to PBMC at least 6 h before LPS challenge . This inhibitory effect of IL-10 was already apparent at the TF mRNA level and was prevented by co-incubation with cycloheximide (20 micrograms/ml) . These data suggest that IL-10 acts via the induction of protein(s) which might interfere with TF gene transcription or mRNA stability . We conclude that the protective effects of IL-10 in endotoxinemia might be related not only to cytokine synthesis blockade but also to inhibition of LPS-induced PCA. Surg Clin North Am, 1993 Oct, 73(5), 1055 - 62 Bacterial, fungal, parasitic, and viral colitis; Schmitt SL et al.; Colitides of bacterial, fungal, parasitic, and viral origin require an organized approach for diagnosis and treatment . Many colitides are transmitted through poor hygienic practices . However, increasing numbers of patients are at risk because of immunocompromise . This population includes the elderly, individuals taking steroids or immunosuppressive agents, and HIV-seropositive individuals . This latter group of individuals is at high risk not only because of immunocompromise but also because of practices such as oroanal intercourse with infected partners . These facts suggest that both the prevalence and the geographic scope of infectious colitides will continue to expand . Undoubtedly, the surgeon will be called on to assist in the diagnosis and management of the complications. Obstet Gynecol, 1993 Oct, 82(4 Pt 1), 550 - 4 A randomized double-blind trial of tinidazole treatment of the sexual partners of females with bacterial vaginosis; Vutyavanich T et al.; OBJECTIVE: To determine whether a single oral 2-g dose of tinidazole for women with clinically diagnosed bacterial vaginosis and their partners increases the cure rates as compared with the same treatment for the female patients alone . METHODS: During a 15-month period, 250 women aged 17-40 years who attended a gynecologic outpatient clinic for abnormal vaginal discharge and/or pruritus vulvae were randomized into two groups . They received a single oral dose of 2 g tinidazole while their partners received either 2 g tinidazole or placebo . Symptomatic improvement and clinical cure rates were assessed at 1 and 4 weeks after treatment . RESULTS: There were no statistical differences (P > .05) in symptomatic improvement, clinical cure rates, or culture results between the groups of women whose partners were treated with either tinidazole or placebo . However, male consorts of women in the tinidazole group experienced side effects more often than those in the placebo group (P = .0006) . CONCLUSION: Routine treatment is not recommended for male partners of women with bacterial vaginosis. J Chir (Paris), 1993 Oct, 130(10), 422 - 5 {Do abuse of nicotine and alcohol have an effect on the incidence of postoperative bacterial infections?}; Stopinski J et al.; In a prospective study we evaluated patients with the diagnosis of 1 . groin hernia (n1 = 57), 2 . gall bladder stones (n2 = 80) and 3 . carcinoma of the colon (n3 = 76) . The whole group included 213 patients who underwent clean, clean-contaminated or contaminated operations . All wound infections and post-operative bacterial infections like pneumonia or urinal infection were registered . The patients were asked for risk factors at the time of hospitalisation . 7.1% of all patients admitted an intake of alcohol of more than 60 g/day and 15.6% of the patients smoked more than 20 cigarettes a day . We found a four times higher risk to get a postoperative infection for patients with an intake of more than 60 g alcohol a day . The rate of infection for smokers of more than 20 cigarettes a day is two times higher than for non smokers or persons who smoke less than 20 cigarettes a day. J Dermatol Sci, 1993 Oct, 6(2), 127 - 33 Activated keratinocytes present bacterial-derived superantigens to T lymphocytes: relevance to psoriasis; Nickoloff BJ et al.; In the past, epidermal keratinocytes were felt to be primarily concerned with the barrier function of skin . During inflammatory and immune-mediated skin diseases, keratinocytes were only portrayed as being passive/inert targets for noxious agents produced by infiltrating leukocytes . This innocent bystander and/or 'brick and mortar' conceptualization of the keratinocyte must now be significantly modified to take into account the growing body of experimental in vitro and in vivo results that substantiate re-classification of keratinocytes as fully fledged members of the immune system (i.e . immunocytes) . Because keratinocytes produce important primary cytokines, adhesion molecules, and mononuclear cell chemotactic factors; as well as functioning as accessory cells for resting T lymphocytes, they can initiate and perpetuate the inflammatory and immunological reactions in the skin which contribute to the pathobiology of psoriasis . This review will emphasize the dynamic contribution that epidermal keratinocytes make to cutaneous immunohomeostasis, with particular focus on the potential role of bacterial derived superantigens and their ability to stimulate resting T cell proliferation when presented by cytokine-activated keratinocytes. J Trop Pediatr, 1993 Oct, 39(5), 284 - 7 Evaluation of CSF variables as a diagnostic test for bacterial meningitis; Deivanayagam N et al.; Specific aetiological diagnosis of bacterial meningitis (BM) in developing countries is often difficult . Frequently, differentiating BM from viral and TB meningitis is not easy . A study was carried out with the easily and quickly performed CSF morphological and biochemical changes as a diagnostic test against the gold standard of CSF culture and/or the latex agglutination test (LAT) . Children between 2 months and 11 years of age, suspected to have acute meningitis, were prospectively recruited . CSF cell count and morphology, Gram stain, culture, and protein and sugar estimations were carried out as per standard procedures . The laboratory personnel were blind to the clinical details and the findings of each other . Diagnosis based on gold standard was possible in 55 out of 114 cases . With CSF polymorphs > 60 per cent and sugar < 50 per cent of blood level as constants, various levels of total cells and protein were considered for their diagnostic properties . The protein level was not useful . We found the best cut-off level of cell count for diagnosis of BM to be 300/mm3, based on the receiver operating characteristics curve, the point of maximum accuracy . These findings were validated by comparing the clinical features, CSF changes and outcome characteristics of non-confirmed cases with the above criteria with the confirmed cases; these were found to be the same except for age.(ABSTRACT TRUNCATED AT 250 WORDS) J Pediatr Gastroenterol Nutr, 1993 Oct, 17(3), 271 - 5 Influence of prenatal corticosteroids on bacterial colonization in the newborn rat; Schiffrin EJ et al.; The interactions between bacteria and the host's intestinal barrier appear to be important regulators of bacterial colonization . In this study we investigated the effect of prenatal corticosteroids, known to accelerate the intestinal maturation of newborn rats, on bacterial colonization in the rat pup . Pregnant rats were treated with either cortisone acetate or normal saline on days 18-21 of gestation and were allowed to deliver spontaneously . The pups, after normal delivery, were sacrificed at different times during the first 10 days of life . The entire small intestine was removed, and each lumen was flushed to exclude nonadherent, transient organisms and homogenized . Tenfold dilutions were plated on horse-blood agar (total bacteria) and MacConkey's medium (gram-negatives) . Quantitation and bacterial typification was determined after 24 h of incubation at 37 degrees C . Total bacteria and gram-negatives found in association with the mucosa were significantly lower in pups prenatally treated with steroids . These changes were not related to any changes in motility or intraluminal digestion . This suggests that the developmental condition of the host's intestinal barrier may be an important regulator of the bacterial microenvironment of the newborn small intestinal mucosa. Immunology, 1993 Oct, 80(2), 333 - 5 Augmented production of interleukin-8 in cerebrospinal fluid in bacterial meningitis; Seki T et al.; Interleukin-8 (IL-8) elaborated by monocytes and endothelial cells is a cytokine which is responsible for adhesion of leucocytes to vascular endothelium and migration of neutrophils into the cerebrospinal fluid (CSF) from the intravascular space . The inflammation in meningitis is elicited by the cytokine release from leucocytes which encounter micro-organisms in the arachnoid or subarachnoid space . In bacterial meningitis, tumour necrosis factor (TNF), IL-1 and IL-6 are produced vigorously, and initiate and augment the inflammation in the central nervous system . In this study, utilizing a quantitative immunometric sandwich enzyme immunoassay, the concentration of IL-8 was investigated in the CSF of patients with bacterial meningitis, patients with aseptic meningitis, and patients with gastroenteritis who served as controls . The IL-8 concentration was markedly higher in the CSF of patients with bacterial meningitis (224 +/- 2.57 pg/ml; mean +/- SD) than in the CSF of patients with aseptic meningitis (less than 30 pg/ml) . The IL-8 level in the CSF of patients with aseptic meningitis did not differ from that in the CSF of the patients with gastroenteritis (less than 30 pg/ml) . The augmented production of IL-8 in CSF may account for the inflammation in bacterial meningitis being more severe than that in aseptic meningitis. J Wildl Dis, 1993 Oct, 29(4), 604 - 7 A serologic survey for some bacterial and viral zoonoses in game animals in the Czech Republic; Hubalek Z et al.; Between 1986 and 1991, sera were collected from 33 roe deer (Capreolus capreolus), 24 red deer (Cervus elaphus), four fallow deer (Dama dama), two mouflon (Ovis musimon), 34 wild boars (Sus scrofa), and 48 hares (Lepus europaeus) shot in two areas of the Czech Republic . Collectively, the sera contained antibodies to Coxiella burnetii (prevalence of 12%), Francisella tularensis (4%), Brucella spp . (2%), Central European tick-borne encephalitis virus (8%), Tahyna (California serogroup) virus (36%), and Calovo (= Batai) virus (23%) . We propose that these mammals may play a role in maintaining natural foci of Q-fever, Tahyna fever and Calovo virus infection. Trends Biochem Sci, 1993 Oct, 18(10), 391 - 5 Rendering a membrane protein soluble in water: a common packing motif in bacterial protein toxins; Parker MW et al.; The recently determined structures of three different protein toxins by X-ray crystallography has unexpectedly revealed a common membrane-insertion domain . This domain consists of an alpha-helical bundle of between seven and ten helices, some of which are hydrophobic . The three toxins, colicin, insecticidal delta-endotoxin and diphtheria toxin are directed towards different hosts, have different killing mechanisms and bear no sequence homology . The observation of a common membrane-insertion domain has implications for the design of therapeutic agents in combating disease. Pediatr Cardiol, 1993 Oct, 14(4), 220 - 2 Parental knowledge of bacterial endocarditis prophylaxis; Cetta F et al.; The aim of this study was to determine parental knowledge of bacterial endocarditis prophylaxis (BEP) . Parents of 135 patients attending a pediatric cardiology clinic in a university center were mailed an eight-question survey pertaining to their knowledge of their child's cardiac disease, medications, and BEP . The patients' cardiac lesions and current medications were verified by a review of clinic and echocardiographic records . Each patient's need for BEP was determined according to American Heart Association (AHA) recommendations . Eighty-four (62%) parents returned complete surveys . The patients' mean age was 5 years with a range of 9 weeks to 19 years . Eighty-two (98%) respondents were high school graduates . Fifty-two (62%) respondents correctly defined endocarditis . Eighty-two (98%) parents knew the correct name of their child's cardiac condition and 27/32 (84%) knew the names of their child's current medications . Only 36/64 (56%) parents of at-risk children knew measures to prevent endocarditis . While most parents know the name of their child's heart lesion and current medications, parental knowledge of endocarditis and BEP was limited . Intensified education and awareness programs are needed in order to prevent potential major morbidity and mortality for pediatric patients with heart disease. Appl Environ Microbiol, 1993 Oct, 59(10), 3513 - 5 Polymerase chain reaction detection of nonviable bacterial pathogens; Josephson KL et al.; Polymerase chain reaction (PCR) methodologies for detection of pathogens in environmental samples are currently available . However, positive amplification products for any set of primers only signal that the appropriate target nucleic acid sequences were present in the sample . The presence of the amplification products does not imply that the target organisms were viable . Here we show that PCR will detect nonviable cells, as long as intact target nucleic acid sequences are available . In an environmental water sample, nucleic acids degraded quickly and were not detectable by PCR after 3 weeks even when stored at 4 degrees C . However, these data show that there is a window of opportunity for PCR analyses to result in false positives with respect to viable cells . We further show that care must be taken in the way samples are stored for future PCR amplifications and that filter sterilization of media is not acceptable for long-term preservation of samples for PCR. Genitourin Med, 1993 Oct, 69(5), 388 - 92 Effect of lactic acid suppositories compared with oral metronidazole and placebo in bacterial vaginosis: a randomised clinical trial; Boeke AJ et al.; OBJECTIVE--To compare the effect of lactic acid locally, metronidazole orally and placebo in women with bacterial vaginosis . DESIGN--Randomised clinical trial . SETTING--30 general practices in the Netherlands . PATIENTS--125 women consulting the general practitioner for symptomatic bacterial vaginosis . MAIN OUTCOME MEASURES--Duration of subjective symptoms, recurrence of symptoms, clinically diagnosed cure, adverse events . RESULTS--Survival analysis showed a significantly faster disappearance of symptoms in the metronidazole category compared with both lactic acid and placebo (p = 0.0005 metronidazole v placebo, p = 0.0002 metronidazole v lactic acid p = 0.6521 lactic acid v placebo {The stratified Mantel Cox test}) . The median duration until absence of symptoms was 21 days for metronidazole and 80 days for placebo . Disappearance of symptoms did not occur in 50% of the lactic acid group in 90 days . Recurrence rates of symptoms were similar over the treatment categories (p = 0.13 metronidazole v placebo and p = 0.12 lactic acid v placebo) . After 2 weeks cure rates (cure defined as less than three of four clinical criteria present) were 83%, 49% and 47% for metronidazole, lactic acid and placebo category respectively . At that time cure rates (cure defined as none of three clinical criteria present) were 10%, 0% and 3% . After four weeks and three months these figures were: 55%, 20%, 20% and 64%, 28%, 28% . No differences in adverse events were found between the three interventions . CONCLUSIONS--Lactic acid suppositories are ineffective, metronidazole capsules are effective on signs and symptoms in bacterial vaginosis . A considerable proportion of the patients recover without active medication. Genetics, 1993 Oct, 135(2), 565 - 74 Cytoplasmic incompatibility and bacterial density in Nasonia vitripennis; Breeuwer JA et al.; Cytoplasmically (maternally) inherited bacteria that cause reproductive incompatibility between strains are widespread among insects . In the parasitoid wasp Nasonia, incompatibility results in improper condensation and fragmentation of the paternal chromosomes in fertilized eggs . Some form of genome imprinting may be involved . Because of haplodiploidy, incompatibility results in conversion of (diploid) female eggs into (haploid) males . Experiments show that bacterial density is correlated with compatibility differences between male and female Nasonia . Males from strains with high bacterial numbers are incompatible with females from strains with lower numbers . Temporal changes in compatibility of females after tetracycline treatment are generally correlated with decreases in bacterial levels in eggs . However, complete loss of bacteria in mature eggs precedes conversion of eggs to the "asymbiont" compatibility type by 3-4 days . This result is consistent with a critical "imprinting" period during egg maturation, when cytoplasmic bacteria determine compatibility . Consequent inheritance of reduced bacterial numbers in F1 progeny has different effects on compatibility type of subsequent male vs . female progeny . In some cases, partial incompatibility occurs which results in reduced offspring numbers, apparently due to incomplete paternal chromosome elimination resulting in aneuploidy. Nervenarzt, 1993 Oct, 64(10), 681 - 4 {Course of bacterial infections in high-dose therapy with clozapine}; Konig F et al.; The influence of neuroleptic drugs on body temperature regulation is well established . Even with atypical neuroleptic drugs such as clozapine, hyperthermia has been observed as well as the malignant neuroleptic syndrome . It is likely that the influence of clozapine affects the immune response . A case is reported in which afebrile angina tonsillaris and endocarditis developed, as a result of high-dosage therapy with clozapine over several years . Besides the special features of the course, differential diagnosis, therapy and possible influence on the body temperature and the immune response are discussed. Plant Mol Biol, 1993 Oct, 23(1), 11 - 21 Increased production of cadaverine and anabasine in hairy root cultures of Nicotiana tabacum expressing a bacterial lysine decarboxylase gene; Fecker LF et al.; Several hairy root cultures of Nicotiana tabacum varieties, carrying two direct repeats of a bacterial lysine decarboxylase (ldc) gene controlled by the cauliflower mosaic virus (CaMV) 35S promoter expressed LDC activity up to 1 pkat/mg protein . Such activity was, for example, sufficient to increase cadaverine levels of the best line SR3/1-K1,2 from ca . 50 micrograms (control cultures) to about 700 micrograms/g dry mass . Some of the overproduced cadaverine of this line was used for the formation of anabasine, as shown by a 3-fold increase of this alkaloid . In transgenic lines with lower LDC activity the changes of cadaverine and anabasine levels were correspondingly lower and sometimes hardly distinguishable from controls . Feeding of lysine to root cultures, even to those with low LDC activity, greatly enhanced cadaverine and anabasine levels, while the amino acid had no or very little effect on controls and LDC-negative lines. Cell Mol Neurobiol, 1993 Oct, 13(5), 483 - 92 VAT-1 from Torpedo synaptic vesicles is a calcium binding protein: a study in bacterial expression systems; Levius O et al.; 1 . Calcium binding properties were examined in VAT-1, an abundant 41-kDa membrane protein expressed in the cholinergic cynaptic vesicles of Torpedo . 2 . An overlay assay, using 45Ca2+ as a tracer, demonstrated the ability of a recombinant VAT-1 produced from the IPTG-inducible pKK223-3 expression vector to bind calcium . 3 . A high yield of recombinant VAT-1 was obtained from the glutathione S-transferase (GST) expression system . The fusion product enabled VAT-1 purification via affinity chromatography . Subsequent cleavage by thrombin resulted in its separation from the GST carrier protein . 4 . A direct Ca(2+)-binding study was performed with purified VAT-1 by a quick-spin column technique, in the presence of 45Ca2+ . Quantitative analysis revealed a 1:1 molar stoichiometry for binding of Ca2+ to VAT-1, with a dissociation constant of 130 microM . 5 . A GST-linked truncated protein consisting of 13 kDa from the VAT-1 carboxy-terminal domain was found to retain the capacity to bind Ca2+ . 6 . A data search for homologies between VAT-1 and known Ca(2+)-binding proteins revealed considerable similarity to members of the annexin family in a 140-amino acid region from the carboxy terminal of VAT-1, which overlaps two tandem Ca(2+)-binding domains of the annexin proteins. Rev Latinoam Microbiol, 1993 Oct-Dec, 35(4), 423 - 32 Computer analysis and modelling of a consensus sequence of bacterial cytochrome oxidase subunit II; Arredondo-Peter R et al.; This work describes the gathering of the reported bacterial cytochrome oxidase subunit II sequences and the construction of the corresponding consensus sequence (COXII CS) . Such sequence was analyzed by computer procedures and a model for the COXII CS was elaborated . The main features of the COXII CS are fully described and the several further applications of this sequence on practical affairs are discussed. Infect Immun, 1993 Oct, 61(10), 4452 - 61 Neither CD14 nor serum is absolutely necessary for activation of mononuclear phagocytes by bacterial lipopolysaccharide; Lynn WA et al.; The stimulation of mononuclear phagocytes by lipopolysaccharide (LPS) is facilitated by the binding of complexes of LPS and LPS-binding protein to CD14 . Although it is clear that CD14 is involved in LPS-induced signaling, other investigators have hypothesized the existence of additional signaling pathways in macrophages . We sought to determine whether CD14-independent pathways of monocyte activation might exist . Washed human mononuclear cells responded with reduced sensitivity to LPS in the absence of serum . Anti-CD14 monoclonal antibody (MAb) inhibited the response to LPS in serum-free conditions, but this was easily reversed at higher concentrations of LPS . We established a human monocytic cell line, designated SFM (derived from THP-1), in serum-free medium to examine LPS responses under defined conditions . Differentiation of SFM cells with 1,25-dihydroxycholecalciferol promoted the expression of abundant cell surface CD14 . Differentiated SFM cells responded to LPS despite the complete absence of serum proteins for > 20 generations of growth . LPS stimulation of differentiated SFM cells was inhibited by anti-CD14 MAbs only when serum was present . In contrast to anti-CD14 MAb, the LPS antagonists lipid IVa and Rhodobacter sphaeroides lipid A inhibited monocyte activation under serum-free conditions, implying that these compounds compete with LPS at a site distinct from CD14 . Undifferentiated SFM cells (expressing minimal CD14) still responded to LPS in serum-free conditions, and anti-CD14 MAb had little inhibitory effect . The addition of purified LPS-binding protein or human serum promoted a CD14-dependent pathway of monocyte activation by LPS in these cells . We conclude that monocytes do not absolutely require serum proteins to be stimulated by LPS and that CD14-independent LPS signaling pathways exist which are inhibitable by lipid IVa and R . sphaeroides lipid A. Curr Opin Genet Dev, 1993 Oct, 3(5), 713 - 8 The retron: a bacterial retroelement required for the synthesis of msDNA; Inouye S et al.; 'Retrons' are bacterial retroelements responsible for the synthesis of msDNA, a hybrid nucleic acid consisting of a single-stranded DNA that is branched out from an internal guanosine of an RNA molecule via a 2',5'-phosphodiester linkage . Retrons are found in a minor population of various bacterial species and are extensively diverse . Two important questions now demanding attention are whether retrons are mobile elements and why are they so diverse? Gene, 1993 Sep 30, 132(1), 83 - 7 Improved bacterial hosts for regulated expression of genes from lambda pL plasmid vectors; Patterson TA et al.; The construction and use of a set of Escherichia coli strains with defective lambda prophages that facilitate expression of genes cloned in lambda pL-plasmid vectors is described . These bacteria allow high and regulated expression of such genes, whereas a kanamycin-resistance marker (KmR) on the prophage allows easy identification and genetic transfer from strain to strain . Optimal conditions for examining gene expression with the pL-vector systems using these strains are discussed. J Chromatogr, 1993 Sep 24, 647(2), 301 - 9 Rapid analysis of enzymatic digests of a bacterial protease of the subtilisin type and a "bio-engineered" variant by high-performance liquid chromatography-frit fast atom bombardment mass spectrometry; van Dongen WD et al.; Amino acid sequencing of a subtilisin-type bacterial protease and a bio-engineered variant was carried out by investigating various enzymatic digests using HPLC-frit fast atom bombardment MS methods . The fast atom bombardment mass spectral data allowed rapid identification of the enzymatically generated peptides and differentiation between both proteins . The feasibility of determining the positions and nature of mutations in the amino acid sequence depends mainly on the size of the peptides containing the modifications. Neurosci Lett, 1993 Sep 17, 160(1), 117 - 9 Bacterial collagenase disrupts extracellular matrix and opens blood-brain barrier in rat; Rosenberg GA et al.; Bacterial collagenase causes hemorrhagic necrosis of brain . We studied the enzyme's effect on blood-brain barrier (BBB) permeability and extracellular matrix (ECM) structure by radiolabeled tracers and electron microscopy . Adult rats had intracerebral injection of bacterial collagenase . Brain uptake from blood of {14C}sucrose was measured in 24 rats 0.5 h to 14 days after injection . 12 rats had ultrastructural studies 1 h after collagenase injection . Brain uptake of {14C}sucrose is maximally increased at 0.5 h, remaining significantly increased for 7 days . Ultrastructurally, some vessels had widening of basal lamina while others had severe disruption of basal lamina with stretching of endothelial cells . We conclude that bacterial collagenase disrupts ECM and opens BBB. Vet Rec, 1993 Sep 11, 133(11), 263 - 6 Sensitivity, specificity and predictive value of blood cultures from cattle clinically suspected of bacterial endocarditis; Houe H et al.; This study investigated the number of blood culture-positive cattle among 215 animals clinically suspected of having bacterial endocarditis . For animals that were necropsied, the sensitivity, specificity and predictive value of the diagnosis of endocarditis were calculated on the basis of the isolation of the causative bacteria from blood . Furthermore, it was investigated whether the glutaraldehyde coagulation time, total leucocyte count, per cent neutrophil granulocytes, pulse rate and duration of disease could help to discriminate endocarditis from other diseases . Among 138 animals necropsied the sensitivity, specificity and predictive value of blood cultivation were 70.7 per cent, 93.8 per cent and 89.1 per cent, respectively . None of the other measurements could be used to discriminate between endocarditis and non-endocarditis cases. Cell, 1993 Sep 10, 74(5), 909 - 17 Folding in vivo of bacterial cytoplasmic proteins: role of GroEL; Horwich AL et al.; A general role for chaperonin ring structures in mediating folding of newly translated proteins has been suggested . Here we have directly examined the role of the E . coli chaperonin GroEL in the bacterial cytoplasm by production of temperature-sensitive lethal mutations in this essential gene . After shift to nonpermissive temperature, the rate of general translation in the mutant cells was reduced, but, more specifically, a defined group of cytoplasmic proteins--including citrate synthase, ketoglutarate dehydrogenase, and polynucleotide phosphorylase--were translated but failed to reach native form . Similarly, a monomeric test protein, maltose-binding protein, devoid of its signal domain, was translated but failed to fold to its native conformation . We conclude that GroEL indeed is a machine at the distal end of the pathway of transfer of genetic information, assisting a large and specific set of newly translated cytoplasmic proteins to reach their native tertiary structures. Circ Shock, 1993 Sep, 41(1), 60 - 5 Bacterial translocation after non-lethal hemorrhage in the rat; Bark T et al.; Translocation of enteric bacteria has been suggested to compromise patients in severe catabolic stress . Mechanisms for this route of infection are not known . In this study, rats were subjected to hemorrhage without reinfusion during 60 min, total blood loss was 3.28 +/- 0.14 ml/100 g BW . Control groups consisted of sham-operated animals without bleeding, and rats not operated at all . The mean number of viable bacteria found in mesenteric lymph nodes (MLN) of bled animals was 168 +/- 45 colony forming units (c.f.u./MLN), significantly higher compared to sham operated (5 +/- 3 c.f.u./MLN) and not operated (0 +/- 0 c.f.u./MLN) controls (P < 0.01) . Cultures from MLN were positive in 7/9 rats after bleeding, in 3/9 of sham operated, and in 0/6 of non-instrumented control animals . No positive blood cultures were isolated . Escherichia coli was the dominant species found in MLN . A biochemical fingerprinting method (the PhP system) was used to identify translocating strains of E . coli among strains found in cecum . The method was also used to compare translocating strains between different animals . Our findings reveal that bacteria translocate to MLN after hemorrhage . Some phenotypes of E . coli strains translocate more frequently than others, suggesting that they have properties facilitating translocation. Protein Sci, 1993 Sep, 2(9), 1461 - 71 Bacterial expression and characterization of the CREB bZip module: circular dichroism and 2D 1H-NMR studies; Santiago-Rivera ZI et al.; In this paper we describe the expression and purification from bacteria of the recombinant basic leucine zipper (bZip) domain of the cAMP response element binding protein, CREB327 . The bZip peptide, CREB259-327, purified to near homogeneity, maintains the sequence-specific CRE site recognition demonstrated by in vitro competition assays . Alkylation of the three cysteine residues of CREB259-327 was employed to prevent aggregation of the peptide due to cysteine oxidation . The Kd of the purified native and modified CREB259-327 for the CRE site was determined by gel retardation assays to be on the order of 10(-7) M . We employed CD spectroscopy to study the folding properties of the native and modified CREB259-327 . The CD analyses of the native/modified CREB259-327 peptide demonstrated a 20% increase in the alpha-helical content upon binding to the cAMP response-element . Only a 5% increase in the alpha-helical content of CREB259-327 is observed upon binding to the AP-1 site . This observation contrasts with CREB from the GCN4 protein (Weiss, M.A., et al., 1990, Nature 347, 575-578) . In addition, the two-dimensional (2D) 1H-NMR studies of the bZip CREB peptide further support the distinct features of the CREB protein, in comparison to GCN4 . Analysis by CD and 2D NMR of the dimerization domain of CREB suggests that the distinct DNA binding characteristics of CREB reside in the basic portion of the bZip module. Proc Natl Acad Sci U S A, 1993 Sep 1, 90(17), 8194 - 8 Beta-galactosidase activity in single differentiating bacterial cells; Russo-Marie F et al.; Myxococcus xanthus strains containing transcriptional fusions to lacZ were analyzed and fractionated by differences in their levels of beta-galactosidase expression . The fluorogenic substrate for beta-galactosidase, fluorescein di-beta-galactopyranoside, was introduced into M . xanthus cells during a rapid decrease in osmolarity of the medium followed by a return to isoosmolarity . Fluorescein, the product of hydrolysis, was retained within the cells and their viability was preserved . Fluorescence increased linearly with time and was proportional to beta-galactosidase activity . beta-Galactosidase expression in most fusion strains, though beginning at different phases of growth or development, was distributed unimodally amongst cells . However, fusion strain Tn5 lac omega 4473 was shown to be heterogeneous at 9 hr of development . It was possible to separate physically cells that expressed beta-galactosidase at a high level from other, still viable, cells with no expression . The approach described here could be adapted to study differentiation in plants and animals as well, where transcriptional fusions and fluorogenic substrates for enzyme probes of gene expression also can be used. Proc Natl Acad Sci U S A, 1993 Sep 1, 90(17), 8159 - 63 Expression of Chinese hamster cAMP-dependent protein kinase in Escherichia coli results in growth inhibition of bacterial cells: a model system for the rapid screening of mutant type I regulatory subunits; Gosse ME et al.; The regulatory and catalytic subunits of cAMP-dependent protein kinase (PKA) were coexpressed within the same bacterial cell using a polycistronic bacterial T7 expression vector encoding Chinese hamster cDNAs for the type I regulatory (RI) and catalytic alpha (C alpha) subunits of PKA . Basal expression of active RI/C alpha holoenzyme in the BL21(DE3) strain of Escherichia coli caused severe growth inhibition resulting in extremely small colony size . Several lines of evidence demonstrate that this growth inhibition requires active PKA subunits and cAMP: (i) this phenotype is dependent on cAMP since it is not seen in a strain lacking adenylyl cyclase activity, but the growth rate of these transformants is slower when exogenous cAMP is added; (ii) normal growth occurs when wild-type RI cDNA is replaced by a mutant RI cDNA encoding a RI protein with reduced cAMP binding; and (iii) the growth-inhibited phenotype of the transformed BL21(DE3) cells requires soluble, active C alpha protein . Holoenzyme expressed in bacteria is activated by cAMP, which stimulates phosphorylation of an endogenous 50-kDa protein that is missing in four host mutants selected for normal growth after transformation with PKA holoenzyme . A mutant RI cDNA library was generated by PCR random mutagenesis and screened by polycistronic expression in BL21(DE3) cells . The RI cDNA sequence from one revertant has base-pair substitutions creating two amino acid substitutions within the cAMP binding sites . The coexpression of the RI/C alpha subunits in BL21(DE3) bacterial cells provides a system for rapidly selecting mutations in the RI subunits of PKA. Clin Immunol Immunopathol, 1993 Sep, 68(3), 283 - 92 Induction of immune-mediated glomerulonephritis in normal mice immunized with bacterial DNA; Gilkeson GS et al.; Normal mice immunized with bacterial DNA produce high titers of anti-DNA antibodies and represent a new model for autoantibody production in systemic lupus erythematosus . To determine whether DNA immunization can also provoke clinical manifestations of lupus, the occurrence of nephritis in immunized mice was assessed and correlated with levels of anti-DNA as well as antibodies to glomerular antigens . BALB/c mice immunized with Escherichia coli single-stranded DNA in complexes with methylated bovine serum albumin in adjuvant showed increased proteinuria compared to control mice immunized with mBSA alone . Furthermore, DNA immunized mice had significantly greater glomerular proliferative changes and immunoglobulin deposition than control mice . In an in vitro assay, sera from DNA immunized mice exhibited greater binding to glomerular antigens than sera from control mice . Compared to sera, renal eluates from DNA-immunized mice were enriched for anti-DNA and glomerular binding activity . These data indicate that immunization of normal mice with E . coli DNA induces an immune-mediated proliferative glomerulonephritis that is likely secondary to the renal deposition of anti-DNA antibodies. J Immunol, 1993 Sep 1, 151(5), 2786 - 93 Synergism between human monocyte chemotactic and activating factor and bacterial products for activation of tumoricidal properties in murine macrophages; Singh RK et al.; Chemotactic factors regulate the recruitment of monocytes-macrophages to inflammatory sites and neoplastic tissues . The purpose of this study was to determine whether MCAF also influences the activation of C3H/HeN macrophages to become tumoricidal . Several metastatic and nonmetastatic clones of the K-1735 murine melanoma cells syngeneic to C3H/HeN mice were transfected with expression vectors containing the human MCAF gene or control DNA . Tumor cells producing high levels of MCAF were significantly lysed by macrophages treated with LPS, whereas parental or control transfected cells were not . Control-activated macrophages incubated with both IFN-gamma and LPS lysed all the melanoma cells regardless of MCAF production . Pretreatment of macrophages with MCAF significantly enhanced their response to low concentrations of LPS, muramyl tripeptide, and a synthetic bacterial LPP, as measured by lysis of murine melanoma cells . These data suggest that in addition to being a chemotactic factor, MCAF can prime macrophages to respond to endotoxins and other bacterial products and therefore may regulate several levels of macrophage-tumor interactions in situ. Obstet Gynecol, 1993 Sep, 82(3), 405 - 10 Efficacy of clindamycin vaginal cream versus oral metronidazole in the treatment of bacterial vaginosis; Fischbach F et al.; OBJECTIVE: To compare the efficacy and tolerance with 2% clindamycin vaginal cream versus oral metronidazole for the treatment of bacterial vaginosis . METHODS: This was a multicenter, randomized, double-blind study in which patients were randomly assigned to one of the following two regimens in a 1:1 ratio: clindamycin phosphate vaginal cream 2% (5 g intravaginally at bedtime for 7 days) plus two placebo capsules (twice a day for 7 days) or metronidazole 500 mg (two 250-mg capsules orally twice a day for 7 days) plus placebo vaginal cream (5 g intravaginally at bedtime for 7 days) . The patients were seen for follow-up at 5-10 days and 25-39 days after completion of therapy . RESULTS: Seven investigators, four in Germany, two in Austria, and one in Switzerland, enrolled 407 patients . Four patients never received either protocol drug, leaving 403 evaluable for safety . Two hundred thirty-four patients were evaluable for efficacy . The analysis for all evaluable patients showed no significant difference between treatment groups . The cure or improvement rate at 1 month after therapy was 83% in the clindamycin group versus 78% in the metronidazole group . The incidence of drug-related adverse medical events was approximately 12% in both groups . CONCLUSIONS: Oral metronidazole and intravaginal clindamycin cream had a similar efficacy of 78 to 83% . Both drugs were tolerated, with vaginal candidiasis developing in 8.5 and 4.7% of the patients in the clindamycin and metronidazole groups, respectively. Cell Signal, 1993 Sep, 5(5), 555 - 64 Evidence for involvement of a GTP-binding protein in activation of Ca2+ influx by epidermal growth factor in A431 cells: effects of fluoride and bacterial toxins; Kuryshev YA et al.; Aluminium fluoride (AlF4-), a G protein activator, was used to study a possible role of G protein in the control of the pathways for Ca2+ influx through plasma membrane of human carcinoma A431 cells . Fluorimetric measurements with the Ca2+ indicator Indo-1 have shown that addition of fluoride induces an increase in concentration of cytosolic free calcium ({Ca2+}in) due to both release of Ca2+ from intracellular stores and Ca2+ influx from the extracellular medium . The cells stimulated by fluoride became unresponsive to subsequent addition of epidermal growth factor (EGF), histamine and bradykinin . The Ca2+ signal induced by fluoride as well as one induced by EGF was inhibited by the pretreatment of cells with protein kinase C activator, phorbol myristate acetate (PMA) . The pretreatment of the cells with pertussis toxin produced no effect on EGF-induced calcium response . In contrast, the pretreatment with cholera toxin (CTX) increased the basal level of {Ca2+}in and abolished the effect of EGF . The effects of CTX could not be reproduced by treating the cells with forskolin or IBMX, agents known to elevate cAMP content in the cell . Patch clamp experiments have shown that fluoride increases the activity of Ca(2+)-permeable channels identical to those activated by EGF from the extracellular side of the membrane {Mozhayeva et al . (1991) J . Membr . Biol . 124, 113-126} . The results obtained suggest the involvement of GTP-binding protein in signal transduction from the EGF receptor to Ca(2+)-permeable channel of plasma membrane in A431 cells. Pancreas, 1993 Sep, 8(5), 597 - 601 Lithostathine, an inhibitor of CaCO3 crystal growth in pancreatic juice, induces bacterial aggregation; Iovanna J et al.; Lithostathine is a pancreatic secretory protein which controls CaCO3 crystal growth in pancreatic juice . Trypsin hydrolysis of the molecule generates two fragments of 11 and 133 amino acids . The N-terminal undecapeptide bears the inhibitory activity for crystal growth . We demonstrate that the C-terminal part of the molecule, which is structurally related to Ca(2+)-dependent lectins, can induce bacterial aggregation . Ca(2+)- and pH-dependent aggregation was obtained for Escherichia coli strain KH 802 and 9 of 19 strains isolated from the predominant flora of human feces . Aggregation of E . coli could be reversed by dilution and bacteria could resume normal growth . Lithostathine is apparently the only component of normal pancreatic juice displaying such activity . Lithostathine is therefore a bifunctional protein which might be involved in the control of the bacterial ecosystem in the intestine. Pancreas, 1993 Sep, 8(5), 551 - 8 Bacterial translocation: a potential source for infection in acute pancreatitis; Gianotti L et al.; Infections from enteric bacteria are a major cause of morbidity and mortality during acute pancreatitis (AP), but the pathways by which these organisms reach distant organs remains speculative . Experiments were conducted to determine if bacterial translocation could be a mechanism for infection during this disease . AP was induced in Lewis rats by i.v . infusion of caerulein (experiment I) or ligation of the head of the pancreas (experiment II) . In a third experiment, rats were gavaged with 1 x 10(8) 14C-radiolabeled Escherichia coli and pancreatitis was induced with caerulein . Results in all three experiments showed that AP increased the number of viable bacteria recovered in peritoneal fluid, mesenteric lymph nodes (MLN), liver, lungs, and pancreas . Radionuclide counting indicated that AP enhanced the gut permeability to 14C E . coli . To estimate the impact of AP on the magnitude of translocation and on the ability of the host to clear bacteria, the nuclide and colony-forming units (CFU) ratios were calculated between animals with and without AP . Blood, peritoneal fluid, and MLN had the highest nuclide ratio . During AP, these tissues may be the principal routes for bacterial spreading from the gut lumen . Peritoneal fluid, pancreas, and lung were the tissues with the highest CFU ratio . Bacterial killing ability of these tissues is likely impaired during AP. G Chir, 1993 Sep, 14(7), 390 - 6 {Secondary bacterial peritonitis: a diagnostic and therapeutic update}; Prisco B et al.; In spite of the availability of new powerful antibiotics, intraabdominal sepsis still has a high mortality rate (20-50%) . With regards to postoperative peritonitis, the referred high mortality rate is due to a late recognition and a consequently late treatment . The causes for late diagnosis are: uncertain clinical picture; misuse of analgesic drugs; reluctance of the surgeon to accept a failure . Moreover, many operated patients are under treatment with antibiotics: their misuse, particularly broad spectrum ones, is responsible for the selection of resistant bacteria . Age is another very significant prognostic factor: mortality rate is constantly higher in elderly subjects than in younger ones . Malnutrition, immunodepression, origin and location of sepsis, concomitant diseases, immunosuppressive treatments, delayed diagnosis, all can significantly affect clinical outcome . Probably the most important and less assessable factor is represented by the surgeon himself with his experience and technical accuracy: any mistake may worse patient's prognosis . Recently, many Authors have stressed the pathogenic relevance of the intestinal mucosa as a barrier, which may influence the clinical course. Zentralbl Bakteriol, 1993 Sep, 280(1-2), 28 - 37 Animal models of bacterial gastritis: the role of host, bacterial species and duration of infection on severity of gastritis; Eaton KA et al.; Gastric bacteria from cheetahs with gastritis were used to inoculate specific-pathogen free kittens and conventional mice . Helicobacter sp . and Gastrospirillum sp . colonized kittens, while only Gastrospirillum sp . colonized mice . In kittens, both bacterial species induced mild lymphofolliclar gastritis which did not change over the course of the 11 months observation period . In mice, Gastrospirillum sp . induced lymphoplasmacytic and follicular gastritis which increased in severity over 6 months and persisted for the 12 month observation period . Gastric ulcers and gastric mucosal hypertrophy were present in chronically infected mice . These results indicate that host but not bacterial factors influence the severity of gastritis, and that in mice, bacterial gastritis increases in severity with time and may lead to gastric ulceration in some individuals. World J Surg, 1993 Sep-Oct, 17(5), 580 - 5; discussion 586 Role of parenteral nutrition in preventing malnutrition and decreasing bacterial translocation to liver in obstructive jaundice; Chuang JH et al.; Surgery in patients with obstructive jaundice is associated with significant infectious complications probably due to impaired immune function and malnutrition . Total parenteral nutrition (TPN) may alleviate malnutrition but may also promote bacterial translocation (BT) from the gut . To elucidate if TPN can prevent malnutrition without promotion of BT in obstructive jaundice, 40 dogs underwent laparotomy for tissue sampling and placement of a central venous line and were allocated into one of four groups: I (PO-control) received dog chow and water ad libitum; II (PO-CBDL) underwent ligation of common bile duct (CBDL) and was fed dog chow; III (TPN-control) received TPN; and IV (TPN-CBDL) underwent CBDL and received TPN . Body weight, blood samples for liver function tests and bacterial culture, and tissues from liver and mesenteric lymph nodes (MLN) for quantitative bacterial culture and for histology were obtained prior to and 2 weeks after the experiment . The incidence of BT to MLN was 40% in the PO-CBDL and TPN-CBDL animals, which was significantly different from the other two groups (0%; p < 0.05) . The incidence of BT to liver was 70% (7/10) in the PO-CBDL animals, which was significantly higher than that in groups I, III, and IV (0%, 20%, 20%, respectively) (p < 0.05) . The PO-CBDL animals showed a significant decrease in body weight and prealbumin compatible with malnutrition, whereas the TPN-CBDL animals showed a significant increase in alkaline phosphatase and a consistent cholestasis on histology . The data suggest that TPN can prevent jaundice-associated malnutrition and decrease BT to liver but should be administered cautiously because it may precipitate cholestasis. J Parenter Sci Technol, 1993 Sep-Oct, 47(5), 192 - 8 Sorption of bacterial endotoxin and retention of bacteria by positively charged membrane filters; van Doorne H; Positively charged membrane filters (Pall, Posidyne) were studied for their ability to retain both bacterial endotoxins and bacteria . Filters (0.2 microns pore size) were challenged with high (up to 5* 10(5) EU) levels of bacterial endotoxin . When the endotoxin was dispersed in water, a more than 10,000 fold reduction of the endotoxin was obtained, whereas the reduction was only a factor 20 when Ringer's solution was used to disperse the endotoxin . Retention of bacteria was dependent on the type of organism, the pore size of the filter and the suspending liquid . Although filters with pore sizes > 0.2 microns were not suitable to be used as a final filter in an aseptic production process, a very reduction in the number of bacteria could be obtained . Possible applications for these filters in the production of sterile pharmaceuticals are discussed. J Burn Care Rehabil, 1993 Sep-Oct, 14(5), 495 - 502 Effects of fluid resuscitation, burn eschar excision, and blockade of afferent pain responses on bacterial translocation and acid-base balance after murine burn injury; Zapata-Sirvent RL et al.; We tested effects of fluid resuscitation, early burn excision/grafting, and blockade of afferent stimuli from the burn wound on bacterial translocation and acid-base balance after murine burn injury . Burn excisions were performed with patients either 15 minutes or 2 hours after burn injury under anesthesia, and excised wounds were immediately closed with murine allograft skin . Twenty-four hours after 25% total body surface area (TBSA) burn injury and 48 hours after 32% TBSA injury, mesenteric lymph nodes were cultured . Incidences of bacterial translocation in 25% and 32% TBSA burns were 31.6% and 68.4% of animals, respectively . Burned animals were in severe shock, and metabolic acidosis reached a nadir 12 hours after burn injury, with base deficit -27.8 +/- 0.6 mEq/L; 5% to 10% of animals died acutely after burn injury . After excision/grafting of burned mice at 2 hours after burn injury, the incidence of bacterial translocation was unchanged (35.7% with 25% TBSA burn, 73.3% with 32% TBSA burn), and mortality did not change . When 32% TBSA excisions were performed exactly 10 minutes after burn injury, four of the 13 mice died within several hours, and five (55.5%) of the nine survivors translocated . Rates of bacterial translocation in mice receiving anesthesia or excision/grafting without burn injury were 15.0% and 20%, respectively (p = NS compared with normal mice) . Subcutaneous implantation of normal or burned skin into normal animals neither elicited shock nor increased the incidence of bacterial translocation . Increasing amounts of fluid resuscitation in the 25% TBSA burn model provided only delayed improvement of acid-base balance; increased amounts of fluid did not decrease bacterial translocation.(ABSTRACT TRUNCATED AT 250 WORDS) J Neurosci Res, 1993 Sep 1, 36(1), 88 - 98 Intracellular distribution of transgenic bacterial beta-galactosidase in central nervous system neurons and neuroglia; Friedrich VL Jr et al.; Bacterial beta-galactosidase is widely used as a marker for gene expression and in cell tracing experiments . In a survey of three transgenic mouse lines expressing beta-galactosidase in the central nervous system (CNS) under the control of different promoters, we find substantial variation in the intracellular distribution of the lacZ protein . In line M beta P5, transgene beta-galactosidase expression is driven by a promoter/enhancer fragment from the oligodendrocyte-specific myelin basic protein gene; however, electron microscopy of histochemically stained preparations reveals transgene expression not only in oligodendrocytes but also in some neurons . Immunofluorescence and immunoperoxidase staining show the beta-galactosidase protein distributed throughout the perikaryal cytoplasm of oligodendrocytes and in processes reaching to myelin sheaths . By contrast, immunoreactive protein appears restricted in neurons to one or a few small perikaryal immunoreactive granules . The granules are visible in the electron microscope as amorphous inclusion bodies of moderate electron density and lack a limiting membrane . Histochemical staining patterns with X-gal and Bluo-gal echoed the protein distribution: diffuse distribution of enzyme protein yielded cells filled with substrate, while punctate enzyme distribution yielded restricted or punctate histochemical staining . Examination of two other lines using different promoter/enhancers to drive expression in the CNS showed both diffuse and punctate beta-galactosidase immunolocalization and histochemical staining . The amount of protein synthesized or other properties, yet unidentified, intrinsic to the target cells may determine the intracellular distribution of beta-galactosidase . In retroviral marking studies, clone members have been identified as those cells filled with X-gal reaction product . This approach may underestimate both clone size and the minimum number of divisions separating the members of each clone. Acta Stomatol Belg, 1993 Sep, 90(3), 181 - 7 ATP content in 3-day-old bacterial plaques with or without a 0.2% chlorhexidine mouth rinse; Vanden Abbeele A et al.; This study describes a method aimed at measuring the bacterial ATP content of dental plaques formed on a polyester support attached on tooth surfaces in the oral cavity . It reports on the variations in ATP content in 3 days old plaques of a well-defined surfaces with or without the use of 0.2% chlorhexidine mouth-wash twice daily . The results, expressed in bacterial equivalents, show a reduction of dental plaque metabolism after 3 days of chlorhexidine rinses to 19.5% of its value in the control tests without mouth-washes . Bacterial metabolism represented 6.05 bacterial equivalents per mm2 in the control group versus 1.18 equivalents in the chlorhexidine test group. Klin Lab Diagn, 1993 Sep-Oct, (5), 22 - 5 {A method of preparing specimens for the electron microscopic study of bacterial fimbrial structures}; Pokhil SI et al.; A method for making bacterial preparations for studies of fimbrial structures by electron microscopy has been developed, making use of impressions of 'young' colonies in agar microscopy . The suggested method involves three steps: cultivation of the examined bacteria in lamellar media till they attain the 'young' age; agar microscopy of the grown culture and making of impressions from the selected sites; contrasting of the objects and drying of the preparations . The new method is simple, economic, highly reliable and yields good results . It maximally rules out the factors conducive to rupture and injury of fimbria, observed in traditional method of making the preparations. Infect Immun, 1993 Sep, 61(9), 3785 - 90 The omp2 gene locus of Brucella abortus encodes two homologous outer membrane proteins with properties characteristic of bacterial porins; Marquis H et al.; In Brucella abortus, a gene encoding a major cell envelope protein, omp2, is duplicated within a short segment of the large chromosomal DNA . Although both genes contain open reading frames, encoding proteins of high identity, expression from only one, omp2b, has been detected in laboratory-grown B . abortus . In the present study, we wished to determine whether omp2b encodes the previously studied Brucella porin and to characterize the omp2a gene product . Experiments were performed with Escherichia coli transformants expressing either omp2a or omp2b . Our results indicated that both gene products localized to the outer membrane of E . coli . Initial rates of transport of {14C}maltose and growth rates in the presence of maltodextrins of defined size indicated an increased hydrophilic permeability of transformants expressing omp2a . These cells were also shown to grow on maltotetraose, a molecule with a molecular mass of 667 Da . Activity consistent with the formation of pores could not be demonstrated in transformants expressing omp2b . However, Omp2b formed oligomers resistant to heat denaturation up to 70 degrees C in sodium dodecyl sulfate buffer, a property characteristic of bacterial porins . Overall, these results suggest that the omp2a gene product has pore-forming activity and that the omp2b gene encodes the previously characterized Brucella porin. FEBS Lett, 1993 Aug 23, 329(1-2), 47 - 50 Yeast mitochondrial ATP-dependent protease: purification and comparison with the homologous rat enzyme and the bacterial ATP-dependent protease La; Kutejova E et al.; Homogenous ATP-dependent protease has been isolated for the first time from mitochondria of yeast Saccharomyces cerevisiae . The enzyme molecule consists of six 120 kDa subunits . It is a serine protease with an absolute ATP requirement for its activity . Basic enzymatic characteristics of the yeast protease are similar to those of the corresponding rat mitochondrial enzyme and of the E . coli protease La . The yeast enzyme immunochemically cross-reacts with the bacterial protease La. Gene, 1993 Aug 16, 130(1), 145 - 50 The Streptomyces coelicolor glnR gene encodes a protein similar to other bacterial response regulators; Wray LV Jr et al.; The Streptomyces coelicolor glnR gene positively regulates the transcription of the glutamine synthetase-encoding glnA gene . The nucleotide sequence of a 1682-bp DNA segment containing glnR was determined . The deduced amino acid sequence of the GlnR protein was found to be similar to the sequence of several bacterial response regulators that are known to function as transcriptional activators . Primer extension analysis of glnR mRNA identified three transcriptional start points (tsp) upstream from the glnR coding sequence. J Biol Chem, 1993 Aug 15, 268(23), 17354 - 61 Distinct mechanisms regulate interstitial collagenase and 92-kDa gelatinase expression in human monocytic-like cells exposed to bacterial endotoxin; Saarialho-Kere UK et al.; We studied the mechanisms that govern the expression of interstitial collagenase and 92-kDa gelatinase in U937 cells, a human monocyte-like cell line, exposed to bacterial lipopolysaccharide (LPS), a potent inducer of metalloproteinase expression . U937 cells were differentiated by phorbol ester (phorbol 12-myristate 13-acetate (PMA)) and, 24 h later, were exposed to LPS for an additional 24 h . Enzyme-linked immunosorbent assay and Northern hybridization showed that PMA mediated an induction of collagenase and markedly stimulated the low basal levels of 92-kDa gelatinase . Subsequent exposure to LPS substantially increased the production of both enzymes . Nuclear runoff assay demonstrated that PMA regulated collagenase and 92-kDa gelatinase transcription . LPS also stimulated collagenase transcription but did not affect transcription of 92-kDa gelatinase . Consistent with the runoff data, the decay rate of collagenase mRNA did not differ between experimental treatments, but the half-life of gelatinase mRNA increased with exposure to LPS . Furthermore, in situ hybridization showed that 92-kDa gelatinase was expressed by all cells whereas collagenase was produced by a subpopulation of cells in both PMA- and PMA/LPS-exposed cultures, and similar findings were seen with LPS-activated human alveolar macrophages . These data indicate that divergent mechanisms control metalloproteinase expression in phagocytic cells and that enzyme production differs among macrophage subpopulations. Arch Biochem Biophys, 1993 Aug 15, 305(1), 30 - 7 Plant dnaj homologue: molecular cloning, bacterial expression, and expression analysis in tissues of cucumber seedlings; Preisig-Muller R et al.; Proteins homologous to the bacterial dnaJ protein have been implicated as molecular chaperones in different compartments of eukaryotes . A plant equivalent is now described in various cucumber tissues . Using a cDNA library constructed from poly(A)-rich RNA of cotyledons of etiolated seedlings and dnaJ-specific probes, we isolated clones encoding full-length cDNAs: clone CSDNAJ-1 contained a 1.7-kb insert with a single open reading frame of 413 amino acid residues encoding a protein of M(r) 46, 012; pCSDNAJ-2 differed from pCSDNAJ-1 by an additional nucleotide and predicted a protein of M(r) 26, 104 . Peptides of similar sizes were observed when the cDNA information was expressed by in vitro transcription and translation or expressed in vivo in bacteria . Using mRNA isolated by hybrid selection for in vitro translation we observed the formation of CSDNAJ-1 protein . Comparisons revealed that the similarity in primary structure between CSDNAJ-1 protein and dnaJ homologues in eukaryotes is most pronounced in the N-terminal region . The high degree of identity (39%) of CSDNAJ-1 protein with the yeast dnaJ homologue YDJ1 protein suggests that csdnaJ proteins assist intracellular protein transfer . The predicted CSDNAJ-1 protein is characterized by a fourfold repetition of the motif CXXCXGX(G) and by a C-terminus -QCAQQ . Analysis of gene expression at the level of mRNA showed that the putative dnaJ protein is expressed in cucumber seedlings in all tissues, but exceedingly high in hypocotyledons and roots . The level of dnaJ mRNA was transiently increased by a factor of 1.5-2.0 when heat shock was applied to the seedlings. Nucleic Acids Res, 1993 Aug 11, 21(16), 3653 - 7 Homologous pairing between single-stranded DNA immobilized on a nitrocellulose membrane and duplex DNA is specific for RecA activity in bacterial crude extract; Bertrand P et al.; Reaction between a circular single stranded and a linear double stranded DNA molecule (ssDNA and dsDNA) provides an efficient system to study recombination mediated by RecA protein . However, classical assays using reaction in solution require highly purified enzymes . This limits biochemical studies of mutant RecA proteins from Escherichia coli or of RecA proteins from other organisms . We describe here an assay that is specific for RecA activity even in bacterial crude extracts . In this assay, the ssDNA is bound to a nitrocellulose membrane, proteins are loaded on this membrane and it is then incubated with a labeled homologous dsDNA . Joint molecules are visualized by autoradiography . We have shown that, despite the reduced mobility of the DNA due to its binding to the membrane, RecA protein is able to promote formation of stable plectonemic joints, in a homology dependent manner . Fourteen other proteins involved in DNA metabolism were checked and did not produce a signal in our assay . Moreover, in Dot blot analysis as well as after native electrophoresis and electrotransfer on a ssDNA coated membrane, production of a signal was strictly dependent on the presence of active RecA protein in the bacterial crude extracts used . We named this assay Pairing On Membrane blot (POM blot). Mol Microbiol, 1993 Aug, 9(3), 533 - 43 NAD+ binding to the Escherichia coli K(+)-uptake protein TrkA and sequence similarity between TrkA and domains of a family of dehydrogenases suggest a role for NAD+ in bacterial transport; Schlosser A et al.; The nucleotide sequence of trkA, a gene encoding a surface component of the constitutive K(+)-uptake systems TrkG and TrkH from Escherichia coli, was determined . The structure of the TrkA protein deduced from the nucleotide sequence accords with the view that TrkA is peripherally bound to the inner side of the cytoplasmic membrane . Analysis by a dot matrix revealed that TrkA is composed of similar halves . The N-terminal part of each TrkA half (residues 1-130 and 234-355, respectively) is similar to the complete NAD(+)-binding domain of NAD(+)-dependent dehydrogenases . The C-terminal part of each TrkA half (residues 131-233 and 357-458, respectively) aligns with the first 100 residues of the catalytic domain of glyceraldehyde-3-phosphate dehydrogenase . Strong u.v . illumination at 252 nm led to cross-linking of NAD+ or NADH, but not of ATP to the isolated TrkA protein. Kansenshogaku Zasshi, 1993 Aug, 67(8), 697 - 703 {Determination of neutrophil function in the respiratory infection by chemiluminescence (CL) . I: Change in neutrophil's CL by chemotherapy against the bacterial pneumonia}; Takeuchi S et al.; We measured neutrophil's CL (CL-index) in 12 patients of the bacterial pneumonia three times per each case: before, during and after chemotherapy . Before the initiation of chemotherapy . CL-index in the six patients remained higher than that in healthy controls, while the remaining six showed lower levels of CL-index compared to the controls . In the 11 cases, their CL-indexes fell to levels lower than those obtained before treatment . Additionally, in the 11 cases their CL-indexes increased after the termination of chemotherapy . Furthermore, in the nine cases the product of the neutrophil number and CL-index was decreased by chemotherapy, and the decrease in the product correlated with improvement in their clinical conditions. Br J Pharmacol, 1993 Aug, 109(4), 987 - 91 Bacterial endotoxin rapidly stimulates prolonged endothelium-dependent vasodilatation in the rat isolated perfused heart; Baydoun AR et al.; 1 . The effects of bacterial lipopolysaccharide (Escherichia coli 0111-B4; LPS) on coronary vascular tone were examined in the isolated perfused heart of the rat . The role of nitric oxide and/or prostaglandin products of the cyclo-oxygenase pathway in mediating the actions of LPS were also investigated . 2 . Coronary vascular tone was raised and maintained by a continuous perfusion of the thromboxane-mimetic U46619 (5 nM) . LPS perfusion (0.1-100 micrograms ml-1) caused a concentration-dependent fall in coronary tone without any significant change in the force of cardiac contractility . 3 . At 5 micrograms ml-1, LPS reduced perfusion pressure by 38 +/- 9 mmHg . This effect was rapid in onset, maximal within the first 5 min and sustained for 90 +/- 10 min (n = 6) . 4 . The vasodilatation induced by LPS was dependent on the presence of an intact endothelium and abolished following endothelial damage caused by air embolism . 5 . NG-nitro-L-arginine methylester (L-NAME; 50 microM) or NG-nitro-L-arginine (L-NOARG; 50 microM) blocked the vasodilatation induced by LPS (5 micrograms ml-1) . The inhibition caused by these arginine analogues was partially reversed by 1 mM L- but not D-arginine . 6 . The vasodilator action of LPS was also completely blocked by the glucocorticoid, dexamethasone (10 microM) but unaffected by indomethacin (10 microM) . 7 . These results suggest that LPS evokes rapid release of nitric oxide (NO) in the microvasculature of the rat isolated heart presumably via activation of the constitutive L-arginine-NO pathway in the endothelium.(ABSTRACT TRUNCATED AT 250 WORDS) Acta Paediatr, 1993 Aug, 82(8), 694 - 8 Lipoprotein alterations in children with bacterial meningitis; Henter JI et al.; Abnormalities in serum lipids, including hypertriglyceridemia, are common during infectious disorders . However, the lipoprotein pattern during infections, particularly in children, has been investigated to only a limited extent . We have monitored alterations in serum lipoproteins in eight children with a severe bacterial infection (meningitis) by a quantitating method measuring cholesterol and triglycerides in each major class . The levels of triglycerides in serum and in low-density lipoproteins were markedly elevated during the infection, whereas the amount of cholesterol in high-density lipoproteins was decreased . The cholesterol to triglyceride ratio was decreased in low-, as well as in high-density lipoproteins . These lipoprotein abnormalities may, at least in part, be explained by a depressed lipolytic activity of lipoprotein lipase, the key enzyme for removal of triglycerides in man . Serum triglycerides and the levels of cholesterol in high-density lipoproteins, as well as the ratio between these parameters, may be used as indicators of inflammatory activity. Appl Environ Microbiol, 1993 Aug, 59(8), 2380 - 7 Characterization of the methanotrophic bacterial community present in a trichloroethylene-contaminated subsurface groundwater site; Bowman JP et al.; Groundwater, contaminated with trichloroethylene (TCE) and tetrachloroethylene (PCE), was collected from 13 monitoring wells at Area M on the U.S . Department of Energy Savannah River Site near Aiken, S.C . Filtered groundwater samples were enriched with methane, leading to the isolation of 25 methanotrophic isolates . The phospholipid fatty acid profiles of all the isolates were dominated by 18:1 omega 8c (60 to 80%), a signature lipid for group II methanotrophs . Subsequent phenotypic testing showed that most of the strains were members of the genus Methylosinus and one isolate was a member of the genus Methylocystis . Most of the methanotroph isolates exhibited soluble methane monooxygenase (sMMO) activity . This was presumptively indicated by the naphthalene oxidation assay and confirmed by hybridization with a gene probe encoding the mmoB gene and by cell extract assays . TCE was degraded at various rates by most of the sMMO-producing isolates, whereas PCE was not degraded . Savannah River Area M and other groundwaters, pristine and polluted, were found to support sMMO activity when supplemented with nutrients and then inoculated with Methylosinus trichosporium OB3b . The maximal sMMO-specific activity obtained in the various groundwaters ranged from 41 to 67% compared with maximal rates obtained in copper-free nitrate mineral salts media . This study partially supports the hypothesis that stimulation of indigenous methanotrophic communities can be efficacious for removal of chlorinated aliphatic hydrocarbons from subsurface sites and that the removal can be mediated by sMMO. Burns, 1993 Aug, 19(4), 302 - 5 Effect of corticotropin releasing factor on acid-base alterations and bacterial translocation in a murine model of thermal injury; Zapata-Sirvent RL et al.; Corticotropin-releasing factor (CRF) is a 41 amino acid polypeptide produced by the hypothalamus which has been shown to decrease inflammation and tissue oedema when administered following burns, cold and acid injuries in some animal models, and to increase mesenteric blood flow . We determined whether systemic administration of CRF to burned mice would decrease metabolic acidosis and protect the gastrointestinal (GI) tract from ischaemic injury leading to bacterial translocation (BT) . Synthetic CRF was administered by intraperitoneal injection in doses of 20 and 200 micrograms/kg to mice immediately following 25 and 32 per cent TBSA burn injuries; the doses were repeated at 8 and 16 h postburn . Severe metabolic acidosis, measured 12 h after burn injury, was not improved in mice which received CRF treatment . Bacterial translocation, measured by quantifying bacteria in mesenteric lymph nodes harvested from animals 48 h postburn, was also not decreased with CRF treatment . CRF does not improve general tissue perfusion nor decrease GI derangements leading to bacterial translocation in this animal model of burn injury. Am J Obstet Gynecol, 1993 Aug, 169(2 Pt 2), 479 - 82 New approaches for the treatment of bacterial vaginosis; Sweet RL; OBJECTIVE: Review alternative approaches to oral metronidazole for the treatment of bacterial vaginosis . STUDY DESIGN: Assessment of clinical trials on the treatment of bacterial vaginosis with regimens other than 5 to 7 days of metronidazole . RESULTS: Single-dose metronidazole, 2 gm orally, is as effective as 5- to 7-day courses of oral metronidazole, with cure rates in the 80% to 90% range . Oral clindamycin results in more than a 90% clinical cure rate . Intravaginal clindamycin cream 2% and intravaginal metronidazole gel 0.75% are associated with clinical cure rates similar to those for oral metronidazole . CONCLUSIONS: Single-dose metronidazole, oral clindamycin, intravaginal clindamycin cream, and intravaginal metronidazole gel are effective and safe alternatives to a 5- to 7-day course of oral metronidazole for the treatment of bacterial vaginosis. Am J Obstet Gynecol, 1993 Aug, 169(2 Pt 2), 446 - 9 Epidemiology of bacterial vaginosis; Mead PB; Studies of the prevalence of and risk factors for bacterial vaginosis are flawed by imprecision in diagnosis, failure to study large and well-characterized populations, selection bias, and failure to correct for confounding