J Biol Chem, 1996 Jul 5, 271(27), 16135 - 8 Purification, cloning, and bacterial expression of retinol dehydratase from Spodoptera frugiperda; Grun F et al.; Anhydroretinol and 14-hydroxy-4,14-retro-retinol, retro-retinoids endogenous to both mammals and insects, act as agonist and antagonist, respectively, in controlling proliferation in lymphoblasts and other retinol-dependent cells . We describe here the identification, purification, cloning, and bacterial expression of the enzyme retinol dehydratase, which converts retinol to anhydroretinol in Spodoptera frugiperda . Retinol dehydratase has nanomolar affinity for its substrate and is, therefore, the first enzyme characterized able to utilize free retinol at physiological intracellular concentrations . The enzyme shows sequence homology to the sulfotransferases and requires 3'-phosphoadenosine 5'-phosphosulfate for activity. Biokhimiia, 1996 Jul, 61(7), 1182 - 8 {Effect of prooxidants on nitric oxide formation in murine liver, treated with bacterial lipopolysaccharide}; Mikoian VD et al.; Coinjection of citrate iron complex (7.5-10 mg iron/kg) with Escherichia coli lipopolysaccharide (LPS) inhibited generation of nitric oxide in the liver of mice caused by LPS-dependent synthesis of inducible NO-synthase (iNOS) . Coinjection of hydroquinone, pyrogallol, or CCl4 with LPS also inhibited NO generation in the liver . Inhibition of LPS-dependent of NO synthesis in the liver by all these agents and iron can be due to their pro-oxidant effects on metabolism of this tissue . These pro-oxidants rapidly activate synthesis of low molecular weight antioxidants; LPS-dependently generated active oxygen forms in the liver of treated animals, which causes iNOS synthesis, are inactivated by the antioxidants; thus, accumulation of active oxygen species is decreased and iNOS synthesis is inhibited. Zh Mikrobiol Epidemiol Immunobiol, 1996 Jul-Aug, (4), 65 - 70 {The lymphocyte phenotype of patients with different forms of diphtheritic infection and of bacterial carriers}; Iushchuk ND et al.; The subpopulation composition of lymphocytes in patients with different forms of diphtheria infection before treatment was studied by the method of laser cytometry with the use of monoclonal antibodies . Statistically significant differences in the characteristics of cell-mediated immunity between groups of diphtheria patients and carriers were obtained . In cases of diphtheria with different degrees of severity an essential decrease in the content of T-lymphocytes and B-lymphocytes was established . The most prolonged changes affected cells carrying differentiating antigens CD3, CD4, CD8, CD72 . Their number increased by day 5 of the disease, while the number of activated cells (CD21 and HLA-DR) progressively decreased . The correlation between the gravity of the course of diphtheria infection and the state of T-cell-mediated immunity was established. Bioorg Khim, 1996 Jul, 22(7), 503 - 9 {Biosynthesis of human calcitonin and mini-proinsulin in bacterial cells in the form of hybrid proteins with corresponding antisense peptides and a metal-binding peptide}; Efimov VA et al.; To verify experimentally the molecular recognition theory, plasmids were constructed that provided the efficient synthesis of hybrid proteins composed of human calcitonin or miniproinsulin, the corresponding antisense peptides, and a histidine-rich metal-binding peptide . A method for isolation of the hybrid proteins by metal-chelating chromatography, cleavage, and renaturation was developed. Mikrobiol Z, 1996 Jul-Aug, 58(4), 67 - 73 {The effect of bacterial polysaccharides on the course of a Mycoplasma infection}; Rudenko AV et al.; Polysaccharides of Clavibacter michiganense have been studied for their effect on Mycoplasma infection . It is established that content of T-lymphocytes truthfully increased in the groups of animals whose PS 1 and PS 2 were administered both with protective and medical purpose as compared with that in a group of animals with natural course of mycoplasmosis . Under these conditions the effect of polysaccharides on T-helpers were not evidently expressed, while they exerted considerable effect on T-suppressors, normalizing their high index in the animals infected by Mycoplasma hominis . Polysaccharides also exerted a normalizing effect on the content of T-killers, whose level in the animals with natural proceeding of mycoplasmosis was considerably decreased . One can observe a decrease of content of B-lymphocytes (which approached the level of their control in a group of conditionally healthy animals) under the effect of polysaccharides as compared to infected animals . Metabolic processes are activated in immunocompetent cells as affected by polysaccharide . Results obtained when studying the effect of polysaccharides on immunologic factors in the animals with mycoplasma infection as well as histomorphological data confirm the above evidences that polysaccharides of C . michiganense are promising immunomodulators. Can J Public Health, 1996 Jul-Aug, 87(4), 264 - 7 {Outbreak of non-bacterial gastroenteritis in a school}; Gaulin C et al.; An outbreak of gastroenteritis occurred in a school affecting more than 30% of its 535 students . An epidemiological survey questionnaire was given to all students as well as staff and maintenance personnel . Stool cultures and electronic microscopy were used to detect the presence of a Norwalk-like virus . Several analyses of water samples were also done . This outbreak occurred simultaneously in the two wings of the school (East and South) . Those who used the East wing were most affected by the disease (RR = 1.45, CI 95%: 1,14-1,85) . There was no indication of food or water supply contamination . A Norwalk-like virus was identified in the stool sample of one child . This along with the clinical characteristics strongly suggested that the pathogen was indeed a Norwalk-like virus . The analysis suggests transmission via contaminated surfaces but also via airborne transport of the infectious agent. J Invest Surg, 1996 Jul-Aug, 9(4), 293 - 303 Bacterial translocation after mesenteric ligation in dogs; Bibbo C et al.; These experiments were designed to determine the relationship between translocation of Escherichia coli and viability of ischemic small bowel . Twenty beagles were gavaged with 14C-labeled E . coli at two time intervals (3 and 24 h) prior to ligation of the blood supply to a 40-cm segment of ileum . Mesenteric lymph node (MLN) biopsies and bacterial cultures of the peritoneal fluid, peripheral arterial blood, and splanchnic venous blood were taken immediately prior to ligation and 24 h later both before and after the ischemic bowel was resected and anastomosed . Biopsies of each resection margin were taken to measure translocation of E . coli into the bowel wall . Several hemodynamic hemodynamic parameters were also measured before and 24 h after ligation . Seven of the 20 dogs died of further bowel necrosis . In survivors A-alpha DO2 was significantly decreased 24 h after mesenteric ligation vs . preligation, whereas in dogs that died DO2 was significantly increased after ligation vs . preligation . The incidence of mesenteric venous cultures positive for E . coli was significantly higher 24 h after ligation vs . preligation . However, there was no correlation between survival and the incidence of positive E . coli cultures in the blood or peritoneal fluid . Mean MLN counts were significantly higher in dogs gavaged at 3 h vs . those gavaged 24 h prior to laparotomy . However, there was no correlation between survival and translocation into either the bowel wall or MLN at either time interval . Viability of ischemic small bowel in this canine model was not affected by translocation of E . coli . Hemodynamic parameters that are altered during the course of sepsis also did not correlate with survival. Gut, 1996 Jul, 39(1), 48 - 53 Allopurinol and glutamine attenuate bacterial translocation in chronic portal hypertensive and common bile duct ligated growing rats; Schimpl G et al.; BACKGROUND: Spontaneous bacterial infections and septicaemia result in morbidity and mortality in patients with portal hypertension and obstructive jaundice . AIM: The aim of this study in rats was to investigate the incidence of bacterial translocation in portal hypertension and obstructive jaundice, and to evaluate the effects of allopurinol and glutamine . METHODS: Rats were subjected to sham laparotomy (SL), portal hypertension (PH) by calibrated stenosis of the portal vein, and common bile duct ligation (CBDL) . Animals of each group were either treated with allopurinol (50 mg/kg twice a week), glutamine (1 g/kg/d), and allopurinol and glutamine . RESULTS: After four weeks, significant bacterial translocation in the untreated PH and CBDL rats occurred . Intestinal mucosal malondialdehyde concentrations (MDA), as an indicator for lipid peroxidation, and myeloperoxidase activity (MPO) released from activated neutrophils were also significantly increased (p < 0.01) . Allopurinol and glutamine in PH and CBDL rats improved bacterial translocation, and decreased MDA and MPO values (p < 0.01) . CONCLUSION: In PH and CBDL rats significant bacterial translocation, ileal mucosal lipid peroxidation, and neutrophil derived MPO activity occurred . Allopurinol and glutamine significantly reduced bacterial translocation, as well as ileal mucosal MDA and MPO activities. J Int Med Res, 1996 Jul-Aug, 24(4), 317 - 24 Bacterial vaginosis in pregnancy and preterm birth: evidence from the literature; Colli E et al.; We review below the evidence from the literature about the relationships between bacterial vaginosis, its treatment and pregnancy outcome . The literature indicates that there is a well-defined association between the presence of bacterial vaginosis during pregnancy and the risk of premature membrane rupture and preterm birth . Less definite is the role of the treatment of such pathology in reducing the frequency of preterm birth and/or premature rupture of membranes . The results of the controlled clinical trials are not entirely consistent . Of the most studied therapies, clindamycin seems to have shown favourable results most consistently but the published data are limited to trials including just a few hundred subjects . Only the availability of further data from controlled clinical trials will clarify the role of such treatment for bacterial vaginosis in pregnancy. Intensive Care Med, 1996 Jul, 22(7), 637 - 43 Impairment of bacterial clearance induced by norepinephrine infusion in rabbits; Koch T et al.; OBJECTIVE: Purpose of the study was to investigate the potential influence of norepinephrine (NE) on immune functions in terms of systemic and organ-specific bacterial clearance in rabbits . DESIGN: To enable quantification of the clearance process, defined numbers of exogenous Escherichia coli (1.3 x 10(8) CFU) were injected intravenously 60 min after starting the NE infusion at a low dose (1 microgram/kg per min, n = 6), causing an increase (30 mmHg) in mean arterial pressure without affecting the oxygen uptake, and at a higher dose (7.5 micrograms/kg per min, n = 6), resulting in a marked decrease (20%) in oxygen uptake, after infusion of NaCl solution (control, n = 6) . In additional experiments (n = 6) NE (1 microgram/kg per min) was tested in endotoxemia induced by simultaneous infusion of endotoxin (40 micrograms/kg per h) . Parameters monitored were arterial pressure, oxygen uptake, and rates of bacterial elimination from the blood . At 180 min after E . coli injection, the animals were sacrificed, and tissue samples of liver, kidney, spleen, and lung were collected for bacterial counts . RESULTS: NE infusion resulted in a dose-dependent prolonged elimination of the injected E . coli from the blood and in significantly higher (p < 0.05) numbers of CFU in liver and lung compared to the controls . Significant impairment of bacterial clearance was found after shock-producing endotoxemia, whereas simultaneous infusion of NE and endotoxin caused only a slightly delayed blood clearance of the injected bacteria . CONCLUSION: NE dose dependently affected bacterial clearance, which might be due to ischemia-derived hypoxic impairment of the phagocytosis and lysis function of the reticuloendothelial system, whereas NE improved elimination of bacteria in a state of endotoxic shock. Zentralbl Bakteriol, 1996 Jul, 284(2-3), 170 - 206 Molecular mechanisms of action of bacterial exotoxins; Balfanz J et al.; Toxins are one of the inventive strategies that bacteria have developed in order to survive . As virulence factors, they play a major role in the pathogenesis of infectious diseases . Recent discoveries have once more highlighted the effectiveness of these precisely adjusted bacterial weapons . Furthermore, toxins have become an invaluable tool in the investigation of fundamental cell processes, including regulation of cellular functions by various G proteins, cytoskeletal dynamics and neural transmission . In this review, the bacterial toxins are presented in a rational classification based on the molecular mechanisms of action. Eur Respir J, 1996 Jul, 9(7), 1523 - 30 The biology of bacterial colonization and invasion of the respiratory mucosa; Wilson R et al.; Despite being regularly exposed to particulate matter during breathing, which contains bacteria from the commensal flora in the nasopharynx and from the environment, the healthy lung is kept sterile by efficient defence mechanisms . Bacterial infections of the respiratory mucosa represent a dynamic interaction, to which both host and bacterial factors contribute . The abnormal host defences associated with chronic respiratory infections (e.g . cystic fibrosis and other forms of bronchiectasis) serve to emphasize their permissive role . The bacteria that cause bronchial infections possess a wide array of potential virulence factors that contribute to their pathogenicity . Many of these factors influence the mucociliary system, an important first-line defence mechanism . The multiplication, spread and persistence of bacteria within the bronchial lumen, and consequent damage to the epithelium, stimulates a chronic inflammatory response, which also impairs mucociliary clearance and damages lung tissue . A greater understanding of host-bacterial interactions during mucosal infections should in the future lead to the development of new therapies and treatment strategies. J Periodontol, 1996 Jul, 67(7), 703 - 16 Correlation between gingivain/gingipain and bacterial dipeptidyl peptidase activity in gingival crevicular fluid and periodontal attachment loss in chronic periodontitis patients . A 2-year longitudinal study; Eley BM et al.; The aim of this study is to determine whether either gingival crevicular fluid (GCF) bacterial gingivain/gingipain or dipeptidyl peptidase (DPP) levels, total activity (TA) and concentration (EC), predict progressive attachment loss (AL) in 75 patients with moderate periodontitis . GCF was collected from 16 molar and premolar mesiobuccal sites and then clinical attachment level (CAL) and probing depth (PD) were measured with an electronic constant pressure probe . Lastly, gingival, gingival bleeding, and plaque indices were scored . Prior to the baseline visit, patients were given basic periodontal treatment after which the above procedures were repeated . In addition, carefully localized radiographs were taken of the test teeth and repeated annually . Patients were then seen every 3 months for 2 years and the clinical measurements repeated at each visit . In 48 patients, 124 AL sites, 91 rapid AL (RAL), and 33 gradual AL (GAL) were detected . Gingivain/gingipain and bacterial DPP levels (TA and EC) at RAL sites were significantly higher (P < or = 0.0001) than at paired control sites at the attachment loss time (ALT) and prediction time (PT) . Mean levels over the study period of both proteases (TA and EC) at GAL sites were significantly higher (P < or = 0.0001) than those at paired control sites . The GCF levels of gingivain/gingipain were always higher than those of DPP . Critical values (CV) of 5 microU/30 seconds (TA) and 30 microU/microL (EC) for both proteases showed high sensitivity and specificity values for TA and EC, which were the same at both ALT and PT . The positive predictive values were higher for gingivain/ gingipain . Mean site levels, over the course of the study, of both proteases (TA and EC) were significantly higher (P < or = 0.0001) at AL, RAL, and GAL sites than non-attachment loss (NAL) sites in AL patients and mean patient levels were significantly higher (P < or = 0.0001) in AL, RAL, and GAL patients than NAL patients . These results indicate that both of these bacterial proteases in GCF may be predictors of periodontal attachment loss. Acta Otolaryngol, 1996 Jul, 116(4), 620 - 6 Bacterial adherence to the surface and isolated cell epithelium of the palatine tonsils; Fredriksen F et al.; Bacterial adherence to the oropharyngeal epithelium is a significant factor in normal microecology, etiopathogenesis of diseases (tonsillitis, gingivitis) and possibly also induction of immune response . Bacterial adhesion to human tonsillar epithelium of whole tonsils and swabs was studied by fluorescence, scanning, and transmission electron microscopy . The epithelial cell borders were well demarcated . On the apical surface of the cells there were irregular microridges . All forms of microscopy visualized epithelial cells with attached bacteria, often forming microcolonies on the free surface . Some bacteria formed excavations on the cell surface . Most attached bacteria were coccoid, but variously sized rods were also visible . In transmission electron microscopy, epithelial cells with intracellular bacteria were regularly observed. Nippon Rai Gakkai Zasshi, 1996 Jul, 65(2), 100 - 5 A serologic Mycobacteria leprae gelatin particle agglutination (MLPA) in the diagnosis of leprosy: comparison with conventional enzyme-linked immunoassay and bacterial index; Agdamag AT et al.; A gelatin particle agglutination test (MLPA) for the detection of anti-phenolic glycolipid-1 (PGL-1) antibodies was compared with the slit skin smear method in the diagnosis of leprosy . MLPA and BI tests showed a good agreement rate of 88.1% and MLPA and ELISA tests showed an excellent agreement rate 96.2% . This MLPA test is simple and reliable, it will be very convenient for the medical practitioners, it would be of great benefit for leprosy patient as well because this test would look like a routine blood examination compared with slit skin smear method which is widely known diagnostic tool for leprosy. Genome Res, 1996 Jul, 6(7), 612 - 9 Bacterial artificial chromosome cloning and mapping of a 630-kb human extrachromosomal structure; Wang M et al.; We have cloned and mapped a circular 630-kb human extrachromosomal structure (termed amplisome) using the bacterial artificial chromosome (BAC) cloning system . Twenty-one BACs were isolated from an amplisome-enriched library by colony hybridization . The insert sizes range from 25 to 143 kb, with an average size of 82 kb . The coverage of the amplisome in clones is approximately 2.7-fold . To construct a physical map of the amplisome, we used three different but complementary methods: hybridization, STS content mapping, and fingerprinting . In addition, we compared the advantages and the drawbacks of these techniques in mapping the amplisomal BACs . The 21 BACs were grouped into two contigs and the two small gaps (3.5 and 26.5 kb) were filled by screening of a human genomic BAC library . The organization of the amplisome revealed by the BAC-based physical map is consistent with the long-range restriction map reported previously . Our results demonstrate that a 630-kb region can be rapidly cloned and mapped into contigs by use of the BAC system . Because of the low frequency (<0.1%) of chimerism and rearrangement, these BAC clones are ready for DNA sequencing and functional analysis. FEMS Microbiol Lett, 1996 Jul 1, 140(2-3), 193 - 8 Bacterial host specificity of Lucinacea endosymbionts: interspecific variation in 16S rRNA sequences; Durand P et al.; Three tropical lucinid clams (Codakia orbiculata, Codakia pectinella and Lucina nassula) from a shallow coastal environment have been studied regarding to their thioautotrophic bacterial endosymbionts . The 16S rRNA genes (rDNA) from these three endosymbionts were amplified using PCR . Phylogenetic analysis by distance matrix and parsimony methods always placed the newly examined symbionts within the monophyletic group composed of symbionts of the bivalve superfamily Lucinacea . A same single 16S rRNA sequence was found in C . orbiculata and C . pectinella and was identical to that found in C . orbicularis and Linga pensylvanica, two other lucinids living in the same type of environment . These data indicate that a same symbiont species may be associated with different host species . Lucina nassula host a symbiont with a distinct 16S rDNA sequence, but very closely related to the former. Eur J Haematol, 1996 Jul, 57(1), 46 - 53 Functional abnormalities in granulocytes predict susceptibility to bacterial infections in chronic lymphocytic leukaemia; Itala M et al.; Leukocyte functions were studied in 22 patients with chronic lymphocytic leukaemia (CLL) and were related to the patients' susceptibility to infections . In CLL patients with a history of infections, compared with CLL patients without infections or healthy controls, there were significant impairments of most granulocyte functions; random migration, N-formyl-methionylleucylphenylalanine (fMLP) and C5a stimulated chemotaxis and chemiluminescence response were decreased . No differences in these functions between CLL patients without infections and healthy controls were observed . Phagocytosis and intracellular killing of granulocytes were intact in all patients with CLL . By univariate analysis, neutrophil count and serum IgG level also predicted susceptibility to infections . By multivariate analyses, granulocyte chemotaxis and chemiluminescence remained as statistically significant predictors of infections . The lymphocyte functions (mitogen-induced lymphocyte proliferation and immunoglobulin production in vitro) were equally impaired in all patients with CLL and differed significantly from the respective functions in healthy control subjects . We conclude that impairments in granulocyte functions contribute to susceptibility to infections in CLL. Am Ind Hyg Assoc J, 1996 Jul, 57(7), 626 - 33 Quantitation of free-living amoebae and bacterial populations in eyewash stations relative to flushing frequency; Bowman EK et al.; This study investigated the concentration of amoebic and bacterial populations in eyewash station water relative to various flushing regimens . Amoebae concentrations averaged approximately 200 amoebae/100 mL in 13 of 15 stations positive for amoebae and consisted of Hartmannella and Acanthamoeba . Bacterial concentrations ranged from 10(0) to more than 10(5) colony forming units per mL . Amoebic concentrations differed notably between stations located in Buildings X and Y (p < 0.0001) . Further study indicated that removal of diffusing screens did not substantially change (p > 0.05) the concentration of amoeba . Amoebic and bacterial concentrations temporarily decreased with the various flushing regimens tested . Lower amoebic concentrations were not sustained by a weekly 3-minute or a monthly 1-minute flushing regimen . However, weekly 3-minute flushes appeared to be more effective in maintaining lowered bacterial concentrations (p < 0.0001). J Neurol Neurosurg Psychiatry, 1996 Jul, 61(1), 57 - 61 Inflammatory cytokines in CSF in bacterial meningitis: association with altered blood flow velocities in basal cerebral arteries; Fassbender K et al.; OBJECTIVE--To investigate the association between release of humoral inflammatory mediators in CSF and blood and alterations of cerebral blood flow in patients with bacterial meningitis . METHODS--Immunomodulatory (interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), and tumour necrosis factor-alpha (TNF alpha)) and vasoactive (thromboxane A, prostacyclin, endothelin-1) molecules of probable or confirmed leucocyte origin were determined in CSF and venous blood from 20 patients with bacterial meningitis, and matched control subjects . Their concentrations were related to the presence of increased blood flow velocities in the middle cerebral arteries, as recorded by transcranial Doppler sonography . RESULTS--Concentrations of proinflammatory cytokines and prostacyclin and leucocyte counts were significantly increased in meningitis, but concentrations of the vasoconstrictors thromboxane and endothelin-1 were not . Patients with high blood flow velocities ( > 140 cm/s) had significantly increased concentrations of IL-1 beta and IL-6 and raised cell counts in CSF . CONCLUSION--The increases of key mediators of inflammation and immunoactivation and of leucocyte count in the CSF of patients with high cerebral blood flow velocities suggest a role of excessive compartmentalised host defence in pathogenesis of disorders of cerebral blood flow in bacterial meningitis. Hum Reprod, 1996 Jul, 11(7), 1575 - 8 Association of bacterial vaginosis with a history of second trimester miscarriage; Llahi-Camp JM et al.; The aim of this study was to determine whether bacterial vaginosis (BV) is associated with a history of recurrent pregnancy loss . A total of 500 consecutive patients attending the Recurrent Miscarriage Clinic were screened for the presence of BV . In women who had had at least one late miscarriage BV was found twice as commonly (27/130; 21%) as in women who had had only early losses (31/370; 8%) (P < 0.001) . The difference was even larger (26 versus 8%) if women who had had term pregnancies were excluded . Moreover, BV was found three times more commonly in Afro-Caribbean women {17 (29%) of 58} than in Caucasian women {36 (9%) of 379} and, in both groups of women, BV was diagnosed at least twice as frequently in those with a history of at least one late miscarriage than in those who had experienced first trimester pregnancy losses only (P < 0 . 001) . The condition occurred twice as often among smokers than non-smokers and, in both groups, it was at least twice as common in women with a history of at least one late miscarriage as in those who had had early pregnancy losses only (P < 0.001) . However, the relationship between BV and smoking was independent of ethnic origin . Women who douched with chloroxylenol were mostly Afro-Caribbean and had BV more than twice as often as women who did not douche. EMBO J, 1996 Jul 1, 15(13), 3219 - 28 Bacterial defense against aging: role of the Escherichia coli ArcA regulator in gene expression, readjusted energy flux and survival during stasis; Nystrom T et al.; Using two-dimensional gel electrophoresis and N-terminal amino acid sequencing analysis, we demonstrate that a mutant of the global regulatory protein ArcA fails to decrease the synthesis of the TCA cycle enzymes malate dehydrogenase, isocitrate dehydrogenase, lipoamide dehydrogenase E3 and succinate dehydrogenase in response to stasis, while the increased production of the glycolysis enzymes phosphoglycerate mutase and pyruvate kinase is unaffected . Microcalorimetric and respiratory measurements show that the continued production of TCA cycle enzymes in the (delta)arcA mutant is manifested as an elevated rate of respiration and total metabolic activity during starvation . The (delta)arcA mutant is severely impaired in surviving prolonged periods of exogenous carbon starvation, a phenotype that can be alleviated by overproducing the superoxide dismutase SodA . In addition, flow cytometry demonstrates that starving (delta)arcA mutant cells, in contrast to wild-type cells, fail to perform reductive division, remain large and contain multiple chromosomal copies . We suggest that the ArcA-dependent reduced production of electron donors and the decreased level and activity of the aerobic respiratory apparatus during growth arrest is an integral part of a defense system aimed at avoiding the damaging effects of oxygen radicals and controlling the rate of utilization of endogenous reserves. J Infect Dis, 1996 Jul, 174(1), 120 - 6 The role of nitric oxide in bacterial meningitis in children; Kornelisse RF et al.; To investigate the role of nitric oxide (NO) in bacterial meningitis, concentrations in serum, cerebrospinal fluid (CSF), or both of the precursor (L-arginine) and degradation products of NO (nitrate, nitrite) and tumor necrosis factor (TNF)-alpha were measured in 35 patients and 30 controls . CSF nitrate levels were significantly elevated, mainly due to increased blood-brain barrier permeability, and are therefore not a good parameter for gauging endogenous NO production in the CSF compartment . CSF NO/nitrite levels were significantly elevated in patients . NO/nitrite levels decreased over time (26%/6 h; P < .001) . CSF levels of NO/nitrite correlated with those of TNF-alpha (r = .55; P = .001) and glucose (r = -.43; P = .02) . CSF levels of L-arginine were lower in patients than in controls (P < .001) . Dexamethasone did not exert a significant effect on NO metabolism . In conclusion, enhanced NO production may contribute to anaerobic glycolysis and neurologic damage in bacterial meningitis. Proc Natl Acad Sci U S A, 1996 Jun 25, 93(13), 6600 - 4 Deciphering the mechanism for the assembly of aromatic polyketides by a bacterial polyketide synthase; Shen B et al.; Aromatic polyketides are assembled by a type 11 (iterative) polyketide synthase (PKS) in bacteria . Understanding the enzymology of such enzymes should provide the information needed for the synthesis of novel polyketides through the genetic engineering of PKSs . Using a previously described cell-free system {B.S . & C.R.H . (1993) Science 262, 1535-1540}, we studied a PKS enzyme whose substrate is not directly available and purified the TcmN polyketide cyclase from Streptomyces glaucescens . TcmN is a bifunctional protein that catalyzes the regiospecific cyclization of the Tcm PKS-bound linear decaketide to Tcm F2 and the 0-methylation of Tcm D3 to Tcm B3 . In the absence of TcmN, the decaketide formed by the minimal PKS consisting of the TcmJKLM proteins undergoes spontaneous cyclization to form some Tcm F2 as well as SEK15 and many other aberrant shunt products . Addition of purified TcmN to a mixture of the other Tcm PKS components both restores and enhances Tcm F2 production . Interestingly, Tcm F2 but none of the aberrant products was bound tightly to the PKS . The results described support the notion that the polyketide cyclase, not the minimal PKS, dictates the regiospecificity for the cyclization of the linear polyketide intermediate . Furthermore, because the addition of TcmN to the TcmJKLM proteins results in a significant increase of the total yield of decaketide, interactions among the individual components of the Tcm PKS complex must give rise to the optimal PKS activity. Proc Natl Acad Sci U S A, 1996 Jun 25, 93(13), 6297 - 301 A bacterial artificial chromosome-based framework contig map of human chromosome 22q; Kim UJ et al.; We have constructed a physical map of human chromosome 22q using bacterial artificial chromosome (BAC) clones . The map consists of 613 chromosome 22-specific BAC clones that have been localized and assembled into contigs using 452 landmarks, 346 of which were previously ordered and mapped to specific regions of the q arm of the chromosome by means of chromosome 22-specific yeast artificial chromosome clones . The BAC-based map provides immediate access to clones that are stable and convenient for direct genome analysis . The approach to rapidly developing marker-specific BAC contigs is relatively straightforward and can be extended to generate scaffold BAC contig maps of the rest of the chromosomes . These contigs will provide substrates for sequencing the entire human genome . We discuss how to efficiently close contig gaps using the end sequences of BAC clone inserts. J Biol Chem, 1996 Jun 21, 271(25), 15285 - 91 Active site phosphorylation of enzyme I of the bacterial phosphotransferase system by an ATP-dependent kinase; Dannelly HK et al.; Enzyme I (EI) of the bacterial phosphoenolpyruvate:glycose phosphotransferase system (PTS) is autocatalytically phosphorylated by P-enolpyruvate . We report here an ATP-dependent kinase (EI-K) from Escherichia coli that reversibly phosphorylates EI at its active site histidine; ATP and EI-K can therefore replace P-enolpyruvate . EI-K contains a bound cofactor that is lost during purification with concomitant loss of activity . NAD+ and NADP+ substitute for the cofactor and restore activity to the apoenzyme, whereas their analogues are inactive . The pyridine nucleotides do not activate EI-K by covalent modification (e.g . ADP-ribosylation), but must be present during the kinase reaction . NADH and NADPH are potent inhibitors of EI-K at all stages of purity, and enzyme activity in a mixture of NAD+ and NADH depends on the ratio of the two pyridine nucleotides . Inhibition is observed with reduced beta-NMN and alpha-NADH, but neither is as effective as beta-NADH . The reverse reaction, the transfer of the phosphoryl moiety from phospho-EI to ADP, also requires NAD+ or NADP+ . In the absence of NAD+ or NADH, {32P}phospho-EI is hydrolyzed to 32Pi, suggesting that EI-K can act as a phospho-EI phosphatase . EI kinase may serve as a link between PTS-driven sugar transport and the electron transport chain. J Immunol Methods, 1996 Jun 10, 192(1-2), 63 - 71 Enhancing immunoelectrochemiluminescence (IECL) for sensitive bacterial detection; Yu H; Immunoelectrochemiluminescence (IECL) as an alternative method versus immunochemiluminescent and immunofluorescent methods can be used for versatile applications in biological agent detection . Although IECL offered high reproducibility and sensitive detection capability for soluble antigens and nucleic acids in aqueous phase, the IECL assays are not optimal and many factors which can affect the IECL performance still remain unclear . Further IECL kinetic studies, improvement of antibody biotinylation, magnetic particle selection and reducing non-specific binding have shown that the enhanced IECL sensitivities (signal to background noise ratios) can be potentially increased at least ten-fold compared to the sensitivities with general IECL assay procedures for bacterial detection. J Immunol Methods, 1996 Jun 10, 192(1-2), 25 - 35 Preparation and characterization of poly-(D,L-lactide-co-glycolide) and poly-(L-lactic acid) microspheres with entrapped pneumotropic bacterial antigens; Kofler N et al.; Poly-(lactide-co-glycolide) microspheres with entrapped antigen have shown considerable promise as controlled release vaccines . To enhance the immunomodulatory effect of LW 50020, a bacterial lysate of seven common respiratory pathogens used perorally as an immunomodulator, we prepared poly-(D,L-lactide-co-glycolide) (PLG) and poly-(L-lactic acid) (PLA) microspheres with entrapped immunomodulator by solvent evaporation or solvent extraction double emulsion techniques . Physical properties, such as particle size, LW 50020 entrapment rate, antigen release patterns and morphological characteristics were investigated . All preparations displayed a high degree of antigen loading up to 95%, whereas size, surface morphology and antigen release patterns were significantly influenced by the method of preparation and the polymer components used . Solvent evaporation microspheres are porous particles from 0.8 micron to 2.0 microns in diameter, that show a rapid antigen release for PLG, and a moderate antigen release for PLA microspheres within 33 days . Solvent extraction microspheres have proven to be particles from 1.1 microns to 5.0 microns in diameter showing a smooth surface and a medium antigen release rate over 33 days . SDS-PAGE and immunoblotting of extracted antigen confirmed that the molecular weight and antigenicity of the immunomodulator remained unaltered by the entrapment procedure. J Immunol Methods, 1996 Jun 10, 192(1-2), 133 - 6 A rapid and sensitive enzyme linked immunofilter assay (ELIFA) for whole bacterial cells; Paffard SM et al.; An improved method is described for the detection of Escherichia coli by an enzyme linked immunofilter assay (ELIFA) using nitrocellulose membrane sandwiched between two 96-well plates . The incorporation of a pumping system permits a continuous flow of reagents and/or wash fluids through the membrane and provides an assay procedure capable of detecting 10(3) bacteria per well within 40 min . Quantitative bacterial detection was based on precipitated chromogen determined by scanning densitometry . The procedure represents a significant improvement in assay time and/or sensitivity over previously described ELIFA and ELISA methods for whole bacterial cells. Br J Hosp Med, 1996 Jun 5-18, 55(11), 685 - 8 Bacterial meningitis; Richardson M; Over the last 10 years, our understanding of bacterial meningitis has improved considerably and new methods of treatment and prevention have been introduced . This review summarizes these advances and discusses more recent developments which will affect the management of bacterial meningitis in the near future. Pulm Pharmacol, 1996 Jun, 9(3), 149 - 56 Effects of bacterial lipopolysaccharides (LPS) and tumour necrosis factor-alpha (TNF alpha) on rat tracheal epithelial cells in culture: morphology, proliferation and induction of nitric oxide (NO) synthase; Freitag A et al.; Rat tracheal epithelial cells were cultured and the effects of LPS and TNF alpha on cell morphology, rate of proliferation and NO synthase activity were studied . NO synthase activity was determined by measuring the accumulation of 3H-L-citrulline during incubation of confluent monolayer with 3H-L-arginine . In untreated cells no significant 3H-L-citrulline formation was detected, and bradykinin and the calcium ionophore A 23187 failed to stimulate 3H-L-citrulline formation excluding a constitutively expressed, calcium-dependent NO synthase activity . After culturing the cells for 18 h in the presence of LPS (10 micrograms/ml) and TNF alpha (500 U/ml) a marked formation of 3H-L-citrulline could be detected, which was largely inhibited by N(G)-monomethyl-L-arginine (L-NMMA) indicating the induction of NO synthase activity which could be prevented by dexamethasone . Exposure of confluent monolayer to LPS and TNF alpha for up to 4 days resulted in a reduction in cell density by 20% within 1 to 2 days and in additional marked changes in cell morphology . The normal honeycomb-like structure of the culture was lost and a considerable number of cells developed "dendritic' outgrowths . These morphological changes as well as the reduction in cell density was attenuated by dexamethasone, but not by the NO synthase inhibitor L-NMMA . The rate of cell proliferation was determined in non-confluent cultures 24 h after passage by determination of the incorporation of tritium into DNA during 24 h of incubation with 3H-thymidine . 3H-thymidine incorporation was reduced by about 40-45% when LPS or TNF alpha was present during exposure to 3H-thymidine, and by about 65%, when LPS and TNF alpha were present in combination . Neither L-NMMA nor dexamethasone significantly affected the 3H-thymidine incorporation nor the inhibitory effects of LPS and TNF alpha . In conclusion, airway epithelial cells are markedly affected by LPS and TNF alpha and the various responses (changes in the cell morphology, inhibition of cell proliferation and induction of NO synthase activity) appear to be caused by different (dexamethasone-sensitive and -insensitive), cellular mechanisms . An enhanced formation of endogenous NO may not be responsible for the observed morphological changes or the inhibition of cell proliferation. Rev Sci Tech, 1996 Jun, 15(2), 603 - 32 Geographical spread of bacterial and fungal diseases of crustaceans; Alderman DJ; The author discusses the way in which bacterial and fungal diseases of crustaceans can be and have been transmitted via commercial movements of stocks . Two classic examples of such diseases are presented, namely gaffkaemia (a bacterial disease of marine lobsters) and crayfish plague (a fungal disease of freshwater crayfish) . In both cases, there is ample evidence that much of the disease spread can be linked to the commercial movement of infected animals caught from infected wild stocks and transported over large distances for wholesale and retail . In the case of crayfish plague, much of the relentless spread of this disease through Europe for nearly 140 years can also be liked to the movement of contaminated fishing equipment without disinfection. J Diarrhoeal Dis Res, 1996 Jun, 14(2), 85 - 9 Hand washing with soap reduces diarrhoea and spread of bacterial pathogens in a Bangladesh village; Shahid NS et al.; Hand washing with soap and water can prevent the spread of diarrhoeal diseases in areas where comparatively costly interventions, such as supply of safe water and improved sanitation, are not possible . In this study, the practice of hand washing with soap and water was instituted in a periurban slum of Dhaka city, and the surveillance for diarrhoea sustained for a one-year period . Rates of primary and secondary attacks were compared to those of a non-intervention area similar in age structure, economic status, education, and other relevant variables . Rectal swabs of cases and contacts established aetiologies . There was a large (2.6 fold) reduction in diarrhoeal episodes in the intervention area during the observation period . Rates of bacterial pathogens were also lower in the intervention area . Significant reduction in diarrhoeal incidences was observed in all age groups for all pathogens except for rotavirus . These observations if implemented as health policy could reduce the spread of diarrhoeal diseases at low cost in high risk areas. Acta Neurol Scand, 1996 Jun, 93(6), 443 - 9 Cerebral hemodynamics in the posterior circulation of patients with bacterial meningitis; Muller M et al.; OBJECTIVES: Stenoses of the middle and anterior cerebral artery (MCA,ACA) are a clinical relevant complication of bacterial meningitis . We examined the involvement of the posterior cerebral (PCA) and of the basilar artery (BA) into the inflammatory process . MATERIAL & METHODS: 39 unselected patients (26 men, 13 women; mean +/- SD age, 49 +/- 17 years) with bacterial (n = 37) and fungal (n = 2) meningitis underwent serial transcranial Doppler sonography recordings of the mean blood velocity (MBV) and the pulsatility index (PI) in the MCA, ACA and PCA, in the BA and in the submandibular internal carotid artery on Days 1, 3, 5, 8, 14 and 21 after admission . The results of the PCA and of the BA were compared to normal reference values and correlated to corresponding neurological signs . The criterion to assume a stenosis was: for the BA an MBV > or = 95 cm/s, for the PCA an MBV > or = 85 cm/s . RESULTS: Compared with the controls MBV in both arteries was normal on Days 1 and 8-21, but significantly increased on Days 3 and 5 . The PI was significantly increased in the BA on all days and in the PCA on Days 1 and 8-21 . Stenoses of the PCA (4 of 43) and of the BA (4 of 32) remained without corresponding neurological signs, and occurred only in patients with two or more narrowed arteries of the anterior circulation (p < 0.05) . CONCLUSION: In bacterial meningitis, hemodynamic disturbances in the posterior circulation are less severe than in the anterior circulation and are clinically well tolerated. Chem Res Toxicol, 1996 Jun, 9(4), 703 - 8 Identification of adducts formed in reaction of adenosine with 3-chloro-4-methyl-5-hydroxy-2(5H)-furanone, a bacterial mutagen present in chloride disinfected drinking water; Munter T et al.; 3-Chloro-4-methyl-5-hydroxy-2(5h)-furanone, MCF, a genotoxic hydroxyfuranone present in chlorine disinfected drinking water, was reacted with adenosine, guanosine, and cytidine in aqueous solutions . HPLC analyses of the reaction mixtures showed that only in the reaction of MCF and adenosine clearly detectable products were formed . The two major products were isolated by C18 column chromatography and characterized by UV absorbance, 1H and 13C NMR spectroscopy, and mass spectrometry . The products were identified as 4-(N6-adenosinyl)-3-formyl-3-butenoic acid (I) and 5-(N6-adenosinyl)-3-chloro-4-methyl-2(5H)-furanone (II) . The yield of I and II in reactions performed at pH 7.4 and 37 degrees C was 0/8% and 0/5%, respectively . Reaction of adenosine with 13C-3-labeled MCF was employed to elucidate the mechanism of formation of I . It was found that the product was formed by a nucleophilic attack of the exocyclic amino group in adenosine on the carbon in the 4-methyl group of MCF . These adducts appear to be novel and not structurally related to those previously identified in the reaction of adenosine with mucochloric acid, another genotoxic hydroxyfuranone. J Biochem (Tokyo), 1996 Jun, 119(6), 1143 - 9 Dual roles of DMPC and CHAPS in the refolding of bacterial opsins in vitro; Sugiyama Y et al.; The bacterial opsins can be refolded to regenerate the chromophore by transfer from SDS to DMPC/CHAPS/SDS mixed micelles in the presence of retinal . A sequential refolding model has been proposed for bacterioopsin {Booth et al . (1995) Nature Struct . Biol . 2, 139-143} . However, the roles of DMPC and CHAPS in the refolding process are not clear . In this study we measured the effects of DMPC and CHAPS on the refolding of bacterial opsins in vitro by CD and fluorescence spectroscopy . In contrast to in experiments in the presence of large amounts of DMPC, the process of retinal binding pocket formation was a rate-determining step in overall chromophore regeneration with relatively low concentrations of DMPC . CHAPS triggered alpha-helix formation and long-range interactions between the helices within 1 s by providing a suitable hydrophobic environment for bacterial opsins . This CHAPS-induced transient molten globule-like structure would be identical to I1 postulated by Booth et al., to which DMPC bound and induced the proper packing of the side chains to form a retinal binding pocket . If DMPC was not present, CHAPS induced another conformation change in bacterial opsins, which led to denaturation . DMPC dependence of chromophore regeneration and the maintenance of the retinal binding pocket suggested that retinal binding pocket formation was part of the large structure changes during stable apoprotein formation. Acta Paediatr, 1996 Jun, 85(6), 670 - 4 C-reactive protein in viral and bacterial gastroenteritis in childhood; Borgnolo G et al.; To differentiate bacterial from viral infections the level of C-reactive protein in serum samples was studied in three groups of children under 5 years of age with gastroenteritis . Of the 53 children with bacterial infection, 41 (77%) had C-reactive protein levels > or = 12 mg1 -1, 32 (66%) > or = 20 mgl-1 and 24 (45%) > or = 35 mgl-1 . Of the 35 patients with viral infection, 4 (11%) had C-reactive protein levels > or = 12 mgl-1, 3 (9%) > or = 20 mgl-1 and 1 (3%) > or = 35 mgl-1 . The best balance between sensitivity and specificity of C-reactive protein was obtained for a cut-off level > or = 12 mgl-1 (sensitivity 77%, specificity 89%) as compared to > or = 20 mgl-1 (sensitivity 58%, specificity 97%) and > or = 35 mgl-1 (sensitivity 44%, specificity 97%) . Our results suggest that the determination of C-reactive protein values may be a useful tool for predicting bacterial gastroenteritis in children. Int J Artif Organs, 1996 Jun, 19(6), 353 - 8 The role of physicochemical properties of biomaterials and bacterial cell adhesion in vitro; Kitano T et al.; This study was undertaken to investigate the physicochemical aspects of the interaction between the surface of biomaterials and bacterial cell membranes in vitro, aimed at studying the mechanisms of bacterial adhesion to biomaterials . Correlations were made between the number of adherent bacterial cells (S . aureus) and each of the calculated components of surface free energy (i.e., dispersion, polarity and hydrogen bond) of biomaterials . The effect of antibodies to cell-adhesion molecules on bacterial adhesion was also studied using monoclonal antibodies to vitronectin receptor, fibronectin receptor and CD44 . This study indicates the polarity component of surface free energy plays a dominant role in the process of bacterial adhesion at least in vitro . The number of cells adherent to materials decreased to 44-73% of the control value in the presence of antibodies tested, showing that cell adhesion molecules affect adherence to biomaterials . Moreover, the results suggested that bacterial adhesion was prevented by specific blockade of cell adhesion molecule receptors. Mol Microbiol, 1996 Jun, 20(5), 903 - 10 Control of bacterial chemotaxis; Eisenbach M; Bacterial chemotaxis, which has been extensively studied for three decades, is the most prominent model system for signal transduction in bacteria . Chemotaxis is achieved by regulating the direction of flagellar rotation . The regulation is carried out by the chemotaxis protein, CheY . This protein is activated by a stimulus-dependent phosphorylation mediated by an autophosphorylatable kinase (CheA) whose activity is controlled by chemoreceptors . Upon phosphorylation, CheY dissociates from its kinase, binds to the switch at the base of the flagellar motor, and changes the motor rotation from the default direction (counter-clockwise) to clockwise . Phosphorylation may also be involved in terminating the response . Phosphorylated CheY binds to the phosphatase CheZ and modulates its oligomeric state and thereby its dephosphorylating activity . Thus CheY phosphorylation appears to be involved in controlling both the excitation and adaptation mechanisms of bacterial chemotaxis . Additional control sites might be involved in bacterial chemotaxis, e.g . lateral control at the receptor level, control at the motor level, or control by metabolites that link central metabolism with chemotaxis. Trends Microbiol, 1996 Jun, 4(6), 226 - 31 Spinning tails: homologies among bacterial flagellar systems; Harshey RM et al.; The bacterial flagellum is a magnificent motion machine that guides the cell in directions dictated by diverse sensory signals . This article reviews the structure and function of this organelle in the context of bacterial variety, and examines a tantalizing connection between motility and virulence that appears to be emerging. Plant Mol Biol, 1996 Jun, 31(3), 553 - 64 Primary structure and expression of plant homologues of animal and fungal thioredoxin-dependent peroxide reductases and bacterial alkyl hydroperoxide reductases; Baier M et al.; Higher plants express genes encoding peroxiredoxins of the two-cysteine type . This is concluded from the isolation of cDNAs from spinach (Spinacia oleracea) and barley (Hordeum vulgare cv . Gerbel) which are homologous to animal, fungal, and bacterial two-cysteine peroxiredoxins . Northern blot analysis indicated the presence of at least one corresponding gene in all angiosperms analyzed suggesting that bas1 is a member of an ubiquitous gene family encoding a protein of fundamental importance in oxidative stress defense also in plants . In barley, expression increased upon application of methyl viologen but was not affected by ozone . mRNA levels increased during deetiolation in the light . Maximal abundance of bas1 transcripts was observed in young developing shoot segments where cell division and elongation take place . Expression was insignificant in roots . The amount of bas1 protein was high in the leaf blade, particularly in etiolated plants, and did not respond to oxidative stress . bas1 protein was not detected in roots . From our data, we suggest that bas1 is an antioxidant enzyme particularly important in the developing shoot and photosynthesizing leaf. Protein Expr Purif, 1996 Jun, 7(4), 389 - 94 Bacterial production and purification of the fish pituitary hormone somatolactin; Pendon C et al.; Somatolactin, a pituitary hormone belonging to the growth hormone/prolactin family, is produced in the intermediate lobe of teleost pituitary . To date, the functions of this new hormone and the target tissues are unknown . A Solea senegalensis somatolactin (ssSL) cDNA has previously been cloned and isolated . Here we have inserted this cDNA into a pET-3a plasmid in order to produce recombinant ssSL in E . coli BL21 (DE3) cells . The protein induced was isolated from inclusion bodies by a solubilization-renaturation procedure originally developed to generate native disulfide bonds, to get putative active proteins . The recombinant somatolactin was further purified to homogeneity by gel filtration on FPLC . The estimated molecular weight of 26 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis agrees well with the molecular mass calculated from the translated cDNA sequence and with native somatolactin (SL) . The recombinant protein showed electrophoretic mobility identical to that of one of the native forms of SL secreted in vitro by cultured pituitaries from sole . Another native SL expressed in S . senegalensis represented a glycosylated modified hormone as shown by N-glycosidase treatment . Further, recombinant SL was recognized by an anti-native SL antibody and used to generate polyclonal sera reactive with the native pituitary hormone . To date, this represents the first recombinant SL protein isolated in sufficient quantities for biophysical and biochemical investigation and for studies on its physiological actions. Br J Nutr, 1996 Jun, 75(6), 881 - 92 Intestinal mucin distribution in the germ-free rat and in the heteroxenic rat harbouring a human bacterial flora: effect of inulin in the diet; Fontaine N et al.; A colorimetric method was used on water-soluble mucin extracted from mucosal scrapings and contents of the caecum and the colon of five germ-free (GF) rats and five heteroxenic (HE) rats harbouring a human flora (GF rats associated with a human flora) . These rats were fed on a diet containing either 100 g sucrose/kg or 100 g inulin/kg . Histological stains, periodic acid-Schiff, alcian blue pH 2.5 and alcian blue pH 0.5 were used to discriminate between neutral, acidic and acidic sulphated mucins respectively . Spectrocolorimetric assays led to a calculated absorbance value for 1 mg of the initial mucin extract . Each mucin type was compared between treatments . The caecal contents of GF rats contained more acidic mucin than sulphomucin, which was present in the same proportion as neutral mucin . Their colonic contents contained more acidic mucins than sulphomucin, which in turn was more abundant than neutral mucin . Their caecal mucosa mucin distribution differed from that of the contents: very little acidic mucin was present and neutral and sulphomucin proportions were of the same order of magnitude . Inulin increased the amount of neutral mucin in the caecal contents and of sulphated mucins in the colonic contents and increased the amounts of neutral and acidic mucins in the caecal mucosa . Mucin distribution in the HE rats was very different from that in the GF rats: the caecal contents contained a high proportion of acidic mucins and very little sulphomucin . The same distribution of mucins was observed in the colonic contents . The caecal mucosa contained less acidic mucin and more sulphomucin than the caecal contents . Inulin decreased acidic mucins and increased sulphated mucins in the caecal contents and increased neutral and sulphated mucins in the colonic contents . Inulin increased sulphomucin in the caecal mucosa and decreased acidic mucin in the caecal and colonic mucosas . The very low amount of mucin that was recovered in the colonic mucosa suggests that, in the presence of the bacterial flora and associated with inulin in the diet, mucin was extensively released from the mucosa to the colonic lumen . This might be related to the bacterial metabolites produced. Enferm Infecc Microbiol Clin, 1996 Jun-Jul, 14(6), 361 - 6 {Bacterial pneumonia as a risk marker for new bacterial diseases in patients with HIV infection}; Teira R et al.; BACKGROUND: Bacterial pneumonias are a main cause of morbidity and mortality among patients infected by the HIV . Furthermore, recent data suggest that, under certain circumstances, they can be prognostic markers . METHODS: The evolution of a cohort of HIV-infected patients, prospectively defined by having suffered a pneumonia, was studied with regards to the occurrence of a new pneumonia or death from a bacterial disease; the evolution of a control cohort of patients matched according to sex, CD4-cell count, previous HIV-disease and zidovudine-treatment was compared to the former cohort by means of survival-analysis techniques . RESULTS: Forty-nine patients in each group were followed up for a total of 1389 and 1893 patient-months respectively . In the first group, 17 new first pneumonia episodes were diagnosed, as compared to 7 in the second group (relative-risk: 3.23, 95% confidence interval: 1.34 to 7.79, p = 0.005) . Eight of the 31 deaths that occurred in the first group were attributed to a bacterial disease, as compared to none of the 24 deaths in the second group (p = 0.007, Fisher's exact test) (in five of those cases, a bacteria was isolated which was thought to be directly related to the death) . CONCLUSIONS: HIV-infected patients who suffer a bacterial pneumonia have an increased risk of suffering new severe bacterial diseases (pneumonia or fatal diseases) as compared to HIV-infected patients with similar prognostic features but no previous episodes of pneumonia. Trends Biochem Sci, 1996 Jun, 21(6), 214 - 9 Quorum sensing: a population-density component in the determination of bacterial phenotype; Swift S et al.; In bacteria, a network of cellular transduction mechanisms integrate signals from the bacterial environment to control gene expression, and thereby the bacterial phenotype . 'Quorum sensing' describes one such signalling mechanism in response to population density . It relies on the accumulation of small extracellular signalling molecules to modulate the transcription of target operons. Neuroscience, 1996 Jun, 72(4), 989 - 97 Activation of serotonergic and noradrenergic neurotransmission in the rat hippocampus after peripheral administration of bacterial endotoxin: involvement of the cyclo-oxygenase pathway; Linthorst AC et al.; An endotoxic challenge produces pronounced effects on the immune, endocrine and central nervous systems . However, information on the brain structures and neurotransmitter systems participating in the physiological responses after stimulation of the immune system is still scarce . Using an in vivo microdialysis method is conscious, freely moving rats, the present study describes the effects of an endotoxic challenge on hippocampal serotonergic and noradrenergic neurotransmission . Rats were equipped with a microdialysis probe in the hippocampus, which enables the stress-free measurement of extracellular concentrations of serotonin, noradrenaline and their respective metabolites 5-hydroxyindoleacetic acid and 3-methoxy-4-hydroxyphenylglycol . The behavioral activity was scored by measurement of the time during which rats were active (locomotion, grooming, eating, drinking) . In the control rats a significant, positive relationship between the behavioral activity and hippocampal extracellular levels of serotonin, noradrenaline and 3-methoxy-4-hydroxyphenylglycol was found . Intraperitoneally injected bacterial endotoxin (lipopolysaccharide; 100 micrograms/kg body weight) increased extracellular concentrations of serotonin, 5-hydroxyindoleacetic acid, noradrenaline and 3-methoxy-4-hydroxyphenylglycol, whereas the behavioral activity was largely reduced, thus disrupting the correlation between behavioral activity and hippocampal levels of serotonin, noradrenaline and 3-methoxy-4-hydroxyphenylglycol . Intraperitoneal pretreatment of rats with the cyclo-oxygenase inhibitor indomethacin attenuated, but did not completely abolish, the endotoxin-induced increases in hippocampal extracellular levels of serotonin, noradrenaline and their metabolites . From these results it may be concluded that the hippocampal serotonin and noradrenaline neurotransmitter systems are part of the brain circuitry responsive to an endotoxic challenge . Moreover, arachidonic acid metabolites seem to represent important, but not the sole, mediators of the endotoxin-induced changes in hippocampal neurotransmission. J Immunol, 1996 Jun 1, 156(11), 4457 - 65 Activation of multiple proline-directed kinases by bacterial lipopolysaccharide in murine macrophages; Sanghera JS et al.; Bacterial LPS stimulation of murine macrophages leads to increased tyrosine phosphorylation and activation of the 42- and 44-kDa mitogen-activated protein kinases (MAPK) and the activation of stress-activated protein kinases (SAPK)/c-Jun N-terminal kinase (JNK) and p38, related to the high osmolarity glycerol protein kinase in Saccharomyces cerevisiae (HOG1) . LPS caused a rapid increase (10 min) in phosphotransferase activity toward myelin basic protein (MBP), a polypeptide that encompassed the first 169 residues of c-Jun fused to gluthathione S-transferase (GST-c-Jun (1-169)) and 27-kDa heat shock protein (hsp27) . MonoQ fractionation of cell extracts resolved phosphotransferase activity peaks toward MBP, GST-c-Jun (1-169), and hsp27, which contained MAPK, SAPK/JNK, and MAPKAPK2, respectively, as indicated by immunoblotting data . In RAW 264.7 macrophages, LPS stimulation of MAPKAPK2, a substrate of p38 HOG1 and MAPK, appeared to occur predominantly via p38 HOG1 and not the MAPK . PMA, which activated the MAPK as potently as LPS, did not strongly activate MAPKAPK2, as assessed by hsp27 phosphorylation . Consistent with p38 HOG1-mediating LPS activation of MAPKAPK2, treatment with LPS, but not PMA, increased the tyrosine phosphorylation of p38 HOG1, a modification known to elevate the enzymatic capacity of this kinase . In LPS-treated cells, the activity of SAPK/JNK was increased 5- to 10-fold, as measured by precipitating SAPK/JNK with Abs or immobilized GST-c-Jun and performing an in vitro kinase assay . In addition, the kinases thought to be upstream of SAPK/JNK, SAPK/ERK kinase 1 (SEK1), and MAPK/ERK kinase kinase 1 (MEKK1), were activated following LPS, but not PMA, exposure (5-fold and 2.5-fold, respectively. J Immunol, 1996 Jun 1, 156(11), 4182 - 90 Roles of proteasomes, transporter for antigen presentation (TAP), and beta 2-microglobulin in the processing of bacterial or particulate antigens via an alternate class I MHC processing pathway; Song R et al.; Latex-OVA and bacteria expressing an OVA fusion protein were processed by macrophages via an alternate class I MHC (MHC-I) processing pathway to present OVA(257-264):Kb . This pathway was resistant to dipeptide aldehyde proteasome inhibitors and brefeldin A, unlike the cytosolic MHC-I pathway . TAP1-/- macrophages exhibited decreases in cell surface peptide-receptive MHC-I and binding of extracellular peptide during transient incubations . This may explain an apparent influence of TAP on alternate MHC-I processing . Alternate MHC-I processing by TAP1-/- cells was enhanced by preincubation at 26 degrees C or with beta 2-microglobulin to increase peptide-receptive MHC-I . Thus, peptides may bind to MHC-I within post-Golgi vacuolar organelles accessible to exogenous beta 2-microglobulin or on the cell surface (following peptide regurgitation). J Surg Res, 1996 Jun, 63(1), 157 - 62 Immediate burn wound excision restores antibody synthesis to bacterial antigen; Yamamoto H et al.; Although burn wound excision and grafting have been shown to improve patient survival, the effects on immune function, especially humoral immunity, are not completely understood . The purpose of this study was to investigate the effect of immediate and early wound excision on antibody synthesis and B-cell proliferation, specifically, antibody response to PGPS, a ubiquitous bacterial cell wall antigen . Thirty-six male BALB/c mice were divided into four groups . Sham mice received no burn, and remaining mice received a 30% body surface area full-thickness burn . Under general anesthesia, excision and grafting was performed either 6 or 72 hr after injury (BE&G6 and BE&G72 groups) . A fourth control group received burn but did not undergo excision and grafting (Burn group) . Splenocytes were isolated 8 days postburn and stimulated with 2.5 microgram/ml lipopolysaccharide . Anti-PGPS IgM, total IgM, and total IgG levels were determined by ELISA . B-cell proliferation, measured by {3H}-thymidine uptake, was expressed as stimulation index . All B-cell functions were significantly suppressed by burn injury . Immediate excision and grafting (BE&G6) restored anti-PGPS IgM synthesis to normal, while nonspecific B-cell functions did not change significantly . Early excision and grafting (BE&G72), however, failed to significantly improve any B-cell functions . Immediate but not early BE&G restored antibody synthesis to the bacterial cell wall antigen (PGPS) . Immediate BE&G may therefore lead to a decrease in bacterial infection after burn injury. Genomics, 1996 Jun 1, 34(2), 213 - 8 Construction and characterization of a human bacterial artificial chromosome library; Kim UJ et al.; We have constructed an arrayed human genomic BAC library with approximately 4x coverage that is represented by 96,000 BAC clones with average insert size of nearly 140 kb . A new BAC vector that allows color-based positive screening to identify transformants with inserts has increased BAC cloning efficiency . The library was gridded onto hybridization filters at high density for efficient identification of BAC clones by colony hybridization . The library was also formulated into characteristic DNA pools to allow for PCR screening of the library for STS content . We have characterized the library mainly by screening with more than 300 different landmarks that include cDNA, STSs, and cosmid clones . We describe methods for using BAC clones and discuss the implications for genome characterization, mapping, and sequencing. Arch Biochem Biophys, 1996 Jun 1, 330(1), 48 - 58 Construction of a human cytochrome P450 1A1: rat NADPH-cytochrome P450 reductase fusion protein cDNA and expression in Escherichia coli, purification, and catalytic properties of the enzyme in bacterial cells and after purification; Chun YJ et al.; A plasmid (pCW) was modified to code for a fusion protein consisting of the complete sequence of human cytochrome P450 (P450) 1A1 (with only the second amino acid changed) in the N-terminal portion connected by a Ser-Thr linker to the portion of rat NADPH-P450 reductase beginning at amino acid 57 . This plasmid was used to express the fusion protein in Escherichia coli DH5alpha cells and the protein was purified from detergent-solubilized bacterial membranes using DEAE and 2',5'-ADP agarose chromatography . The purified fusion protein catalyzed benzo{a}pyrene 3-hydroxylation, 7-ethoxyresorufin O-deethylation, and zoxazolamine 6-hydroxylation . Catalytic activity was not increased in the presence of added NADPH-P450 reductase, cytochrome b5, or phospholipid . The fusion protein could also transfer electrons to cytochromes c and b5 but not P450 lA2 . The same oxidation products of benzo{a}pyrene were formed with the purified fusion protein and the fusion protein functioning in bacterial cells . The catalytic activity of the human P450 1A1 fusion protein toward several substrates is markedly less than that of a similar fusion protein constructed with rat P450 1A1, in line with the reported differences in catalytic activities of the rat and human P450 1A1 enzymes . The purified fusion protein also oxidized (+)- and (-)-benzo{a}pyrene 7,8-dihydrodiols and eight aryl and heterocyclic amines to genotoxic products, in the absence of added NADPH-P450 reductase . The demonstration of catalytic activities of the human fusion protein within bacterial cells suggests the prospect of utilizing such cellular systems for production of human P450 metabolites. J Pediatr, 1996 Jun, 128(6), 765 - 9 Granulocyte colony-stimulating factor as a marker for bacterial infection in neonates; Kennon C et al.; OBJECTIVE: To evaluate granulocyte colony-stimulating factor (G-CSF) as an early marker of bacterial or fungal infection in neonates . STUDY DESIGN: We measured G-CSF levels in infants of varying gestational and postnatal ages . We separated the infants into three groups: group 1, positive bacterial or fungal blood culture result; group 2, negative blood culture result but evidence of clinical sepsis; and group 3, negative blood culture result and no or weak evidence of sepsis . Comparison of mean G-CSF levels by group was accomplished by an analysis of variance . RESULTS: One hundred seventy-six evaluations for sepsis were done for 156 infants with gestational ages ranging from 24 to 43 weeks; 50% of these infants were less than 35 weeks of gestational age . The mean G-CSF levels of groups 1 and 2 were significantly higher than those of group 3 . The mean G-CSF level of each group was 2278 pg/ml (group 1), 1873 pg/ml (group 2), and 280 pg/ml (group 3) (p < 0.001) . On the basis of a cutoff level of 200 pg/ml, the sensitivity of the test was 95%, specificity 73%, positive predictive value 40%, and negative predictive value 99% . CONCLUSION: G-CSF levels represent a sensitive marker of infection in neonates of all gestational ages. J Infect Dis, 1996 Jun, 173(6), 1498 - 502 Interleukin-10 and soluble tumor necrosis factor receptors in cerebrospinal fluid of children with bacterial meningitis; Kornelisse RF et al.; The antiinflammatory mediators interleukin (IL)-10 and soluble tumor necrosis factor (TNF) receptors p55 (sTNFR-55) and sTNFR-75 in cerebrospinal fluid (CSF) from 37 children with bacterial meningitis were studied . CSF concentrations of IL-10, sTNFR-55, and sTNFR-75 and of the proinflammatory cytokines TNF-alpha, IL-6, and IL-8 were markedly elevated and were, with the exception of the sTNFRs, significantly higher in CSF than in serum . CSF concentrations of sTNFR- 55 and sTNFR-75 were only associated positively with IL-10 levels . CSF glucose levels correlated highly with levels of IL-10, sTNFR-55, and sTNFR-75 and weakly with TNF-alpha and IL-6 . Cytokine levels in CSF decreased rapidly, while sTNFR levels remained elevated for at least 24 h. Biochim Biophys Acta, 1996 May 28, 1311(3), 150 - 4 Relevance of the arginine transport activity to the nitric oxide synthesis in mouse peritoneal macrophages stimulated with bacterial lipopolysaccharide; Shibazaki T et al.; Transport of arginine and production of nitrite have been investigated in mouse peritoneal macrophages stimulated with bacterial lipopolysaccharide (LPS) . The arginine transport activity was induced by LPS at very low concentration (maximally induced at 1 ng/ml), whereas much higher concentration of LPS was required for the induction of nitrite production . Arginine was more concentrated in the cells when its transport activity was induced . Lysine, which is a competitive inhibitor of the transport of arginine, neutralized the concentrative effect of the induced transport activity and thus inhibited the nitrite production . Induction of the arginine transport activity seems to be prerequisite to the enhanced synthesis of nitric oxide in activated macrophages. J Biol Chem, 1996 May 24, 271(21), 12394 - 400 Insertion of the polytopic membrane protein MalF is dependent on the bacterial secretion machinery; Traxler B et al.; We examined the dependence of protein export and membrane protein insertion on SecE and SecA, two components of the secretion (Sec) apparatus of Escherichia coli . The magnitude of the secretion defect observed for signal sequence-containing proteins in cells depleted of SecE is larger and more general than that in many temperature- or cold-sensitive Sec mutants . In addition, we show that the proper insertion of the polytopic MalF protein (synthesized without a signal sequence) into the cytoplasmic membrane is also SecE-dependent . In contrast to an earlier study (McGovern, K., and Beckwith, J . (1991) J . Biol . Chem . 266, 20870-20876), the membrane insertion of MalF also is inhibited by treatment of cells with sodium azide, a potent inhibitor of SecA . Therefore, our data strongly suggest that the cytoplasmic membrane insertion of MalF is dependent on the same cellular machinery as is involved in the export of signal sequence-containing proteins . We propose that the mechanism of export from the cytoplasm is related for both signal sequence-containing and cytoplasmic membrane proteins, but hydrophobic membrane proteins such as MalF may have a higher affinity for the Sec apparatus. J Biol Chem, 1996 May 24, 271(21), 12356 - 63 Potential induced redox reactions in mitochondrial and bacterial cytochrome b-c1 complexes; Tolkatchev D et al.; Purified cytochrome b-c1 complexes from beef heart mitochondria and Rhodobacter sphaeroides were reconstituted into potassium-loaded asolectin liposomes for studies of the energy-dependent electron transfer reactions within the complexes . Both complexes in a ubiquinone-sufficient state exhibit antimycin-sensitive reduction of cytochromes b (both low and high potential ones) upon induction of a diffusion potential by valinomycin in the presence of ascorbate . Addition of N,N,N',N'-tet-ramethyl-p-phenylenediamine (TMPD) to the ascorbate-reduced potassium-loaded asolectin proteoliposomes resulted in reduction of cytochrome b262 . Upon addition of valinomycin, the induced diffusion potential caused a partial reoxidation of cytochrome b562 and partial reduction of cytochrome b566 in beef heart cytochrome b-c1 complex in the presence of antimycin and/or myxothiazol . Surprisingly, when ubiquinone-depleted beef heart cytochrome b-c1 complex liposomes were treated under the same conditions, no cytochrome b566 reduction was observed but only the oxidation of cytochrome b562, and the oxidation was not oxygen-dependent . We explain this effect by b566, iron-sulfur protein short-circuiting under these conditions, assuming that both antimycin and myxothiazol markedly affect subunit b conformation . The electrochemical midpoint potential of heme b566 appears to be significantly higher than that of heme b562 in the presence of myxothiazol, which cannot be accounted for only by the potential-driven electron transfer between these two hemes plus the shift in chemical midpoint potentials caused by myxothiazol . A model for energy coupling consistent with structural findings by Ohnishi et al . (Ohnishi, T., Schagger, H., Meinhardt, S . W., LoBrutto, R., Link, T . A., and von Jagow, G . (1989) J . Biol . Chem . 264, 735-744) is presented . This model is a compromise between pure "redox-loop" and pure "proton-pump" mechanisms . Reoxidation of high potential heme b is observed in an antimycin- or antimycin plus myxothiazol-inhibited, ascorbate plus TMPD-prereduced R . sphaerodies b-c1 complex, upon membrane potential development, suggesting that a similar electron transfer mechanism is also operating in the bacterial complex. FEBS Lett, 1996 May 20, 386(2-3), 252 - 4 Direct involvement of hydrogen peroxide in bacterial alpha-hydroxylation of fatty acid; Matsunaga I et al.; We have reported that fatty-acid alpha-hydroxylase partially purified from Sphingomonas paucimobilis required NADH and molecular oxygen . In this study, we found that the reaction was greatly inhibited by catalase . Glutathione and glutathione peroxidase also inhibited alpha-hydroxylation, but superoxide dismutase and mannitol did not . Replacement of NADH and molecular oxygen by hydrogen peroxide increased the alpha-hydroxylation activity . In the presence of hydrogen peroxide, molecular oxygen was not required for the activity . These findings suggest that hydrogen peroxide was essential for bacterial alpha-hydroxylase. Biochem J, 1996 May 15, 316 ( Pt 1), 209 - 15 Regulation of the mouse inducible-type nitric oxide synthase gene promoter by interferon-gamma, bacterial lipopolysaccharide and NG-monomethyl-L-arginine; Weisz A et al.; Cytokines and bacterial lipopolysaccharides (LPSs) stimulate nitric oxide production in macrophages by inducing transcription of the gene coding for the inducible isoform of nitric oxide synthase (iNOS) . We have cloned the mouse iNOS gene promoter and analysed its structural features and its response to interferon-gamma (IFN-gamma) and Escherichia coli LPS in RAW 264.7 mouse macrophage-like cells . Transcription of a recombinant reporter gene including the promoter and 4 kb of its 5'-flanking DNA, linked to the bacterial chloramphenicol acetyltransferase (CAT) reporter gene, is stimulated by IFN-gamma and, more efficiently, by LPS upon transient transfection in RAW 264.7 cells . Two upstream DNA regions are required for maximal promoter activation of LPS: the first maps between positions -1541 and -775 and the other between -420 and -47, with respect to the major transcriptional start site of the iNOS gene . The upstream-most region also mediates promoter trans-activation by IFN-gamma . As reported earlier for transcription of the endogenous iNOS gene, combined stimulation of RAW 264.7 cells with IFN-gamma and LPS results in lower activation of the transfected promoter, when compared with LPS alone . NG-Monomethyl-L-arginine, a competitive inhibitor of nitric oxide synthase activity, enhances iNOS gene mRNA induction and promoter activation by IFN-gamma and LPS, indicating that nitric oxide can influence negatively the reponsiveness of this gene to inducers . These results suggest the possibility of a negative regulatory feedback exerted by iNOS on the transcriptional activation of its own gene. FEBS Lett, 1996 May 13, 386(1), 5 - 10 Bacterial and plant-produced scFv proteins have similar antigen-binding properties; Bruyns AM et al.; A gene encoding a single-chain variable (scFv) antibody fragment was expressed as a cytoplasmic and endoplasmic reticulum-targeted protein in transgenic tobacco plants . In both cases, the scFv accumulated up to 0.01% of total soluble protein (TSP) . The same scFv fragment was also produced in the periplasm of Escherichia coli . Measurement of the affinity by ELISA indicates that the affinity of the bacterially made scFv is about 80-fold lower than that of the parental Fab fragment . The results suggest that the affinity of the plant-produced scFv fragments is reduced to a similar extent, implying that all the plant-produced scFv fragments are antigen binding. J Immunol Methods, 1996 May 10, 191(1), 39 - 48 A method for titration of inhibiting antibodies to bacterial immunoglobulin A1 proteases in human serum and secretions; Reinholdt J; Bacterial IgA1 proteases specifically cleave IgA1, including S-IgA1, molecules into Fab alpha and Fc alpha fragments . Hereby these enzymes interfere with the protective functions of antibodies belonging to this isotype . Antibodies inhibiting IgA1 proteases have been detected in humans, but the titration of such antibodies is a matter of methodological concern . Because human serum and secretions contain IgA1 substrate, it is impossible to provide uniform substrate conditions for samples of IgA1 protease incubated with inhibitors differing in their origin and state of dilution . This study demonstrates that such variations in substrate are not prohibitive for a reliable titration of inhibiting antibodies . This was evident from experiments demonstrating that the variations do not interfere with the quantification of residual IgA1 protease activity provided the activity is measured in terms of the proportion of IgA1 substrate cleaved during incubation . Proportions of cleaved IgA1 were measured by exploiting the differential reactivity of cleaved and intact IgA1 molecules in an ELISA using anti-Fc alpha and enzyme-conjugated anti-light chain antibodies for catching and development, respectively . A protocol for the titration of IgA1 protease-inhibiting antibodies based on this ELISA is described . By application of the protocol to chromatographic fractions of saliva, IgA1 protease-inhibiting activity was found to co-purify with salivary S-IgA. Med J Aust, 1996 May 6, 164(9), 546 - 8 Managing HIV . Part 5: Treating secondary outcomes . 5.16 HIV and bacterial infections; Allworth AM et al.; Patients with HIV infection have a high risk of bacterial infection . A high index of suspicion for common and less common bacterial infections is needed, together with a flexible approach to diagnosis and therapy . Many infections are characterized by relapse after therapy; long-term treatment is frequently required. J Mol Biol, 1996 May 3, 258(2), 270 - 85 Mutations in motB suppressible by changes in stator or rotor components of the bacterial flagellar motor; Garza AG et al.; Five proteins (MotA, MotB, FliG, FliM and FliN) may be involved in energizing flagellar rotation in Escherichia coli . To study interactions between the Mot proteins, and between them and the three Fli proteins of the switch-motor complex, we have isolated extragenic suppressors of dominant and partially dominant motB missense mutations . Four of the 13 motB mutations yielded partially allele-specific suppressors . Of the suppressing mutations, 57 are in the motA gene, eight are in fliG, and one is in fliM; no suppressor was identified in fliN . The prevalence of suppressors in fliG suggests that FliG interacts rather directly with the Mot proteins . The behaviour of cells in tethering and swarm assays indicates that the motA suppressors are more efficient than the fliG or fliM suppressors . Some of the suppressing mutations themselves confer distinctive phenotypes in motB+ cells . We propose a model in which mutations affecting residues in or near the putative peptidoglucan-binding region of MotB misalign the stator relative to the rotor . We suggest that most of the suppressors restore motility by introducing compensatory realignments in MotA or FliG. Trends Genet, 1996 May, 12(5), 192 - 6 Complications and implications of linear bacterial chromosomes; Chen CW; The emergence of linear bacterial chromosomes has overthrown the dogma of universal circularity of the bacterial chromosomes, and posed mechanistic and evolutionary implications not previously anticipated. Indian Pediatr, 1996 May, 33(5), 373 - 6 C-reactive protein as an indicator of complications in bacterial meningitis; Singh UK et al.; OBJECTIVE: To determine the value of serum C-reactive protein (CRP) estimation in the follow-up of bacterial meningitis (BM) . DESIGN: Longitudinal follow-up . SETTING: Urban hospital . METHODS: Sequential serum CRP estimation was done in 50 healthy children and in 100 children with BM . Serial serum CRP value was correlated with the clinical picture in BM . RESULTS: Serial serum CRP in 14 patients with complicated BM was significantly different than the 72 uncomplicated BM cases . CONCLUSIONS: Serial determination of serum CRP may be a cheap, simple and reliable prognostic indicator in BM. J Neuroimmunol, 1996 May, 66(1-2), 65 - 70 Interleukin-1 receptors type I and type II in the mouse brain: kinetics of mRNA expressions after peripheral administration of bacterial lipopolysaccharide; Gabellec MM et al.; The expression of transcripts for Interleukin-1 (IL-1) type I and type II receptors (IL-1R1, IL-1R2) was investigated in the mouse brain and spleen using reverse transcription-polymerase chain reaction techniques under basal conditions and following injection of endotoxin (LPS, i.p., 4 mg/kg) . Under basal conditions, mRNAs for both receptor types were found in various parts of the brain, in pituitary as well as in spleen . Following LPS stimulation, mRNA expressions were increased in all studied tissues . IL-1R1 mRNAs were predominant in the brain and pituitary while, IL-1R2 mRNAs were more abundant in the spleen . The maximal quantity of transcripts (IL-1R1, IL-1R2) was obtained 6 h after LPS injection in all studied tissues . The decrease to basal level was observed within 48 h in the brain . In the spleen, IL-1R1 mRNAs remained elevated 48 h after LPS while IL-1R2 mRNAs had already reached basal level . These results indicate a LPS-induced stimulation of IL-1 receptors mRNAs in the brain and a differential expression of IL-1R1 and IL-1R2 transcripts in brain and immune tissues. Minerva Stomatol, 1996 May, 45(5), 189 - 96 {The correlation between an increase in subgingival temperature and the concentration of volatile fatty acids of bacterial origin present in periodontal pockets}; Eftimiadi C et al.; The results obtained in this study indicate that the mean subgingival temperature measured in periodontal patients is significantly higher compared to that obtained from healthy subjects . The increase in subgingival temperature observed in periodontal patients, is proportional to the concentration of short-chain fatty acids (SCFA) present in the gingival fluid . The highest temperature values were observed when suppuration and high levels of SCFA were both present in the same site . Theoretical considerations are presented which suggest a possible role of metabolically activated neutrophils in the local temperature increase . In any case, considering that the local increase in temperature is associated with an inflammatory process and that SCFA are bacterial "toxins" capable of inhibiting the immunological host response, the results presented in this study agree with the hypothesis that SCFA may be involved in the pathogenesis of chronic adult periodontitis. Nutrition, 1996 May, 12(5), 303 - 15 Bacterial products and the control of ingestive behavior: clinical implications; Langhans W; Bacterial products such as lipopolysaccharides (LPS) and muramyl peptides are delivered in the course of infections . They trigger the host's acute phase responses to bacterial infections and are probably involved in the accompanying hypophagia because LPS and muramyl dipeptide (MDP, the minimal immunologically active muramyl peptide) reduce food intake after parenteral administration in animals . LPS and MDP inhibit feeding synergistically through separate but interacting mechanisms . The hypophagic effects of LPS and MDP are presumably mediated by the combined actions of interleukin-1, tumor necrosis factor, and other cytokines . More work is required to understand the interactions between these cytokines, and between bacterial products and cytokines, before cytokine antagonists can be used for treatment of the hypophagia during bacterial infections . As the hypophagia seems to be an early mechanism of host defense, a treatment should be carefully considered . If an intervention is indicated because of a patient's poor condition, inhibitors of eicosanoid synthesis and glucocorticoids may hold more promise for therapy because such substances block LPS and MDP hypophagia . Although LPS can reduce food intake by direct action on the brain, presently available evidence indicates that systemic LPS acts primarily in the periphery to generate a neural signal that is transmitted to the brain and inhibits feeding through the vagus . The exact site where LPS acts on peripheral nerves remains to be identified . LPS hypophagia is conditionable, but conditioning cannot solely account for LPS hypophagia under most test conditions . Whether MDP hypophagia is also conditionable and mediated by vagal afferents is not yet known . All in all, the putative mediators and mechanisms of LPS and MDP hypophagia suggest some options for a treatment of the hypophagia during bacterial infection, but present knowledge about the mechanisms and interactions of the involved substances is still fragmentary and requires further investigation. J Pediatr Surg, 1996 May, 31(5), 665 - 9 Acute hypoxia does not increase bacterial translocation in newborn rabbits; Urao M et al.; PURPOSE: We have previously demonstrated that spontaneous bacterial translocation (BT) occurs in newborn rabbits and correlates strongly with small bowel colonization (BC) . Birth stress, specifically hypoxia, is believed to increase this pathologic process and thus lead to sepsis . This study investigated the relationship between BT and acute hypoxia in newborn rabbits . METHODS: Four hundred seventeen rabbit pups (aged 0, 2 to 4, 6, and 28 days) were divided into four groups according to the type of hypoxic stress: 9% O(2) for 1 hour, 9% O(2) + 12% CO(2) for 1 or 4 hours, and 21% O(2) (control animals) . The animals were killed 1.5 or 20 hours after the stress . Sterile specimens of mesenteric lymph nodes (MIN), spleen, liver, small bowel, and large bowel were incubated aerobically at 37 degrees C for 24 hours in thioglycolate broth, and subsequently plated on both MacConkey and Colistin Naladixic Acid media . After 24 hours, the growth on both plates was recorded . X(2) analysis was used, and P values of less than .05 were considered significant . RESULTS: BC of the small bowel and BT to the MLN were low in the first 4 days of life in the hypoxic groups (range, 0% to 21% BC, 0% to 6% BT) and the control group (range, 4% to 30% BC, 3% to 12% BT) . After an increase in BC at 6 days of age, the rate of BT increased to 25% to 29% in control animals . The rate of BT in the hypoxic groups (25%) did not differ significantly from that of the controls (P > .05) . Additionally, killing at 20 hours (v 1.5 hours) was not associated with an increase in the incidence of BT . None of the stress groups had a significant increase in BT compared with the controls . Importantly, although 4 hours of 9% O(2) + 12% CO(2) resulted in a 30% mortality rate, the incidence of BT was no higher than that of the control animals (13% v 29%; P > .05) . CONCLUSION: Severe hypoxic stress in newborn rabbits does not increase the incidence of BT . Because the incidence of BT correlates with that of BC, and because BC is the same in the control and hypoxic animals, the sepsis observed in hypoxic newborns probably is not related to an increased incidence of BT. Am J Otol, 1996 May, 17(3), 375 - 85 A human temporal bone study of acute bacterial meningogenic labyrinthitis; Merchant SN et al.; It is well established that sensorineural hearing loss (SNHL) is an important sequela of acute bacterial meningitis . Previous human temporal bone histopathologic studies have suggested that such hearing loss is due to labyrinthitis . This study involved a detailed and systematic evaluation of the auditory and vestibular end-organs in 41 human temporal bones from patients with acute bacterial meningitis, aimed at describing the spectrum of histopathologic changes within the labyrinth, ascertaining likely routes for spread of infection from the meninges to the inner ear, and comparing the data from humans with those described in a rabbit model of meningogenic labyrinthitis . Our study revealed the following: (a) Suppurative labyrinthitis occurred in 20 (49%) bones . Of these 20 bones, the cochlea was affected in all, whereas the vestibular organs were involved in 10 . Eosinophilic staining of inner ear fluids without the presence of inflammatory cells (so-called "serous" labyrinthitis) occurred in 14 of the remaining 21 bones . This staining occurred primarily within the vestibular system . Its significance and pathogenesis remains unknown; (b) Sensory and neural structures of the inner ear appeared intact in the majority of specimens, including bones with suppurative labyrinthitis and those with eosinophilic staining of inner ear fluids . This finding raises the possibility of preventing or reversing SNHL by therapeutic intervention . Spiral ganglion cells were severely degenerated in 12% of bones, indicating a retrocochlear site of hearing loss in addition to the cochlea . This subset of patients may perform poorly after cochlear implantation; (c) It has been traditionally assumed that irreversible and permanent SNHL is caused by suppurative labyrinthitis, whereas reversible SNHL is caused by serous labyrinthitis . Our findings question the validity of these assumptions; (d) The data were consistent with the hypothesis that both the cochlear modiolus and cochlear aqueduct can serve as potential pathways for spread of infection from the meninges to the inner ear; (e) There were many similarities in the histopathology of the inner ear in humans when compared with the rabbit model of meningogenic labyrinthitis . A notable difference was that the cochlear aqueduct appeared to be the sole pathway for spread of infection in the rabbit, whereas in the human, both the modiolus and aqueduct were possible pathways. Anal Chem, 1996 May 1, 68(9), 1646 - 50 Homogeneous bioluminescence competitive binding assay for folate based on a coupled glucose-6-phosphate dehydrogenase--bacterial luciferase enzyme system; Huang W et al.; A homogeneous bioluminescence competitive binding assay for folate was developed by using a coupled enzyme system of glucose-6-phosphate dehydrogenase (G6PDH) and bacterial luciferase . A highly substituted G6PDH-folate conjugate was prepared by employing an N-hydroxysuccinimide/carbodiimide method . Folate binding protein inhibits the activity of the conjugate . In the presence of folate, there is a competition between folate and the G6PDH-folate conjugate for the binding site of the folate binding protein, and the activity of the conjugate is recovered . Thus, the concentration of folate can be related to the activity of the G6PDH-folate conjugate, which is directly related to the bioluminescence produced by the coupled enzyme reaction . Using this assay, dose-response curves with a detection limit of 2.5 x 10(-8) M folate were obtained, which is an improvement of an order of magnitude with respect to an assay that monitors G6PDH activity spectrophotometrically . The assay was validated using vitamin tablets and a cell culture medium. Arch Oral Biol, 1996 May, 41(5), 393 - 400 A comparison of cysteine and serine proteinases in human gingival crevicular fluid with tissue, saliva and bacterial enzymes by analytical isoelectric focusing; Gazi MI et al.; Gingival crevicular fluid (GCF) contains several different proteinase activities and the study sought to clarify their sources . Gingival tissue and GCF were collected from chronic periodontitis patients . Gel-filtration chromatography of crude tissue extracts yielded cathepsin B and tryptase fractions sensitive to cysteine and serine proteinase inhibitors, respectively . Cell sonicates of suspected periodontal pathogens were prepared from broth cultures of reference strains . Of these, Porphyromonas gingivalis showed much the strongest activity and this had an effector response consistent with the metal-dependent cysteine proteinase described by others . Banding patterns in GCF, tissue and bacterial samples were compared on substrate-impregnated overlay membranes applied to isoelectric focusing gels . On Z-Val-Lys-Lys-Arg-AFC overlays, GCF had bands corresponding to tissue cathepsin B and the enzyme from P . gingivalis, though a contribution from Treponema denticola could not be ruled out . Use of D-Val-Leu-Arg-AFC overlays showed GCF activity similar to tissue tryptase . In GCF there were additional bands that did not correspond to any tissue or bacterial samples and on Z-Ala-Ala-Lys-AFC overlays these closely resembled activity in parotid saliva . The results confirmed that GCF contains tissue cathepsin B and tryptase, while the apparent presence of enzymes from P . gingivalis and possibly T . denticola is consistent with previous reports linking activity to these organisms . The saliva bands demonstrated that contamination of GCF may occur despite rigorous collection procedures. Allergy Asthma Proc, 1996 May-Jun, 17(3), 137 - 42 Chronic cutaneous bacterial hypersensitivity; Backman KS et al.; Chronic cutaneous bacterial hypersensitivity (CCBH) is a syndrome of recurrent indolent inflammatory lesions that are caused by a hypersensitivity reaction to skin flora . It can cause significant physical and social distress to those affected, who often endure years of nondiagnostic evaluations and ineffective treatments . We report two additional cases of this syndrome . Skin testing demonstrated IgE to bacterial antigens in both patients, and ELISA demonstrated both IgE and IgG to bacterial antigens in one patient . Both patients improved rapidly and dramatically with oral corticosteroid therapy, although they failed to achieve a lasting remission, as had been seen in previous cases . These represent the fourth and fifth reported cases of CCBH . Despite failure to achieve remission, these patients demonstrate the effectiveness of an inexpensive and safe prednisone regimen in controlling this physically and socially destructive disease. Can J Public Health, 1996 May-Jun, 87(3), 208 - 11 {Outbreak of non-bacterial gastroenteritis in a school}; Gaulin C et al.; An outbreak of gastroenteritis occurred in a school affecting more than 30% of its 535 students . An epidemiological survey questionnaire was given to all students as well as staff and maintenance personnel . Stool cultures and electronic microscopy were used to detect the presence of a Norwalk-like virus . Several analyses of water samples were also done . This outbreak occurred simultaneously in the two wings of the school (East and South) . Those who used the East wing were most affected by the disease (RR = 1.45, CI 95%: 1,14-1,85) . There was no indication of food or water supply contamination . A Norwalk-like virus was identified in the stool sample of one child . This along with the clinical characteristics strongly suggested that the pathogen was indeed a Norwalk-like virus . The analysis suggests transmission via contaminated surfaces but also via airborne transport of the infectious agent. J Periodontol, 1996 May, 67(5), 497 - 505 Analytical performance of an immunologic-based periodontal bacterial test for simultaneous detection and differentiation of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia; Snyder B et al.; The analytical performance of a membrane-based immunoassay for the simultaneous detection and differentiation of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia (including Prevotella nigrescens) was investigated . Positive reactions were observed for 71 of 71 reference strains and recent oral isolates of A . actinomycetemcomitans, P . gingivalis, and P . intermedia . No cross-reactivity was observed with 39 other common oral and environmental species . The specificity of the test was unaffected by the presence of potential oral interferents including whole blood, white blood cells, mucin, saliva, toothpastes, and oral rinses . A proficiency test by dental professionals using a standardized set of unknown simulated samples yielded a sensitivity of 97% (116/120) and a 100% specificity (240/ 240) . An additional group including dental professionals and high school students was shown to be 99% proficient (1385/1397) in distinguishing proper from improper test function when processing control samples with normal test devices and devices with simulated error conditions . Comparisons to a culture standard for 104 subgingival plaque samples collected from 26 adult periodontitis patients yielded > 98% specificity for each of the test bacteria . In addition, the detection threshold for the test was determined to be equivalent to 10(4) cultivable test bacteria when compared to the culture standard . The data indicate that this membrane immunoassay is a valid and easy-to-use method for the detection of A . actinomycetemcomitans, P . gingivalis, and P . intermedia in subgingival plaque, at levels above the detection threshold of the test. Gut, 1996 May, 38(5), 784 - 7 Obstructive jaundice causes reduced expression of polymorphonuclear leucocyte adhesion molecules and a depressed response to bacterial wall products in vitro; Plusa S et al.; BACKGROUND--Obstructive jaundice is associated with an increased incidence of infection and endotoxaemia, which may result from impaired host immunity . Neutrophil adhesion to vascular endothelium is a key part of the inflammatory response . AIMS--To investigate neutrophil adhesion molecule expression and activation in obstructive jaundice . PATIENTS--Nine adult patients with obstructive jaundice and 11 control subjects . METHODS--The expression of the neutrophil adhesion receptors L-selectin, CD11a, CD11b, CD11c, and CD15 was determined using flow cytometry . CD11b expression in response to stimulation with fMLP and endotoxin was measured . RESULTS--The basal expression of L-selectin, CD11a, and CD15 was significantly decreased in jaundiced patients (p < 0.05) and the expression of CD11b in response to stimulation with fMLP and endotoxin was significantly impaired in the jaundiced group . Endotoxin stimulation without plasma did not reverse the impaired response showing that it is not caused by endotoxin inactivation by plasma proteins . CONCLUSIONS--Neutrophils from patients with obstructive jaundice show decreased adhesion receptor expression and an impaired response to stimulation with bacterial products . This cellular dysfunction may be responsible for the high incidence of septic complications in these patients. Obstet Gynecol, 1996 May, 87(5 Pt 1), 656 - 60 The preterm prediction study: fetal fibronectin, bacterial vaginosis, and peripartum infection . NICHD Maternal Fetal Medicine Units Network; Goldenberg RL et al.; OBJECTIVE: To determine the relation between vaginal and upper genital tract infection and cervical-vaginal fetal fibronectin levels . METHODS: We screened 2899 women at ten centers every 2 weeks from 23-24 to 30 weeks' gestation for cervical and vaginal fetal fibronectin . A positive test was defined as a level of at least 50 ng/mL . The relation between a positive test and bacterial vaginosis at 23-24 weeks and clinical or histologic chorioamnionitis at delivery plus neonatal sepsis was determined . RESULTS: Fetal fibronectin was present in 4.0% of cervical and/or vaginal samples at 23-24 weeks and was nearly twice as common in women with bacterial vaginosis . Adjusting for the presence of bacterial vaginosis, race, and parity, women positive for fetal fibronectin were much more likely to have clinical chorioamnionitis (mean +/- standard deviation gestational age 30.6 +/- 4.1 weeks), with an odds ratio of 16.4 and 95% confidence interval of 7.1-37.8, and neonatal sepsis (6.3 and 2.0-20.0, respectively), than those who were fetal fibronectin-negative . A positive cervical fetal fibronectin test was a better predictor of clinical chorioamnionitis and neonatal sepsis than was a vaginal test or a combination of vaginal and cervical tests . Among 40 women who delivered before 32 weeks and had placental histology available for evaluation, ten had a positive cervical and/or vaginal fetal fibronectin test before delivery; all ten had histologic evidence of chorioamnionitis, compared with only 13 of 30 women (43%) who were fetal fibronectin-negative (P = .02) . CONCLUSION: Women with bacterial vaginosis were more likely to have a positive fetal fibronectin test than uninfected women . Women with a positive fetal fibronectin test who delivered before 32 weeks' gestation all had evidence of histologic chorioamnionitis . Women positive for fetal fibronectin also had a 16-fold increase in clinical chorioamnionitis and a sixfold increase in neonatal sepsis . There is strong evidence that upper genital tract infection and cervical and/or vaginal fetal fibronectin are closely linked. Transgenic Res, 1996 May, 5(3), 193 - 201 Improved metabolic action of a bacterial lysine decarboxylase gene in tobacco hairy root cultures by its fusion to a rbcS transit peptide coding sequence; Herminghaus S et al.; The gene of a bacterial lysine decarboxylase (ldc) fused to a rbcS transit peptide coding sequence (tp), and under the control of the CaMV 35S promoter, was expressed in hairy root cultures of Nicotiana tabacum . The fusion of the ldc to the targeting signal sequence improved the performance of the bacterial gene in the plant cells in many respects . Nearly all transgenic hairy root cultures harbouring the 35S-tp-ldc gene contained distinctly higher lysine decarboxylase activity (from 1.5 to 30 pkat LDC per mg protein) than those which had been transformed with constructs in which the gene had been directly cloned behind the CaMV 35S promoter . The higher enzyme activity led to the accumulation of up to 0.7% cadaverine on a dry mass basis . In addition, part of the cadaverine pool was used for increased biosynthesis of anabasine, an alkaloid which was hardly detectable in control cultures . The best line contained anabasine levels of 0.5% dry mass, which could be further be enhanced by feeding of lysine. Laryngorhinootologie, 1996 May, 75(5), 293 - 300 {Interdisciplinary therapeutic concept in severe bacterial infections of the central respiratory tract in childhood}; Damm M et al.; BACKGROUND . Apart from all advances made in the management of central airway infections, Acute Epiglottitis (AE) and Bacterial Tracheitis (BT) continue to be causes of life-threatening airway obstruction in children . The aim of this retrospective study was to evaluate deficiencies in the diagnostical protocol, to clarify the role of airway endoscopy in acute childhood stridor, and to identify current reasons for fatalities in these diseases . MATERIAL . In the observation period between 1980-92, we found 12 patients suffering from BT and 21 from AE managed in close cooperation of the involved disciplines at the pediatric intensive care unit of the University of Cologne . RESULTS . Laryngoscopy with fiberoptic or small rigid endoscopes allowed in awake cooperative children accurate diagnose of AE, and the exclusion of supraglottic inflammation in BT without complications . Furthermore, additional endoscopic information of the degree of inflammation was helpful in the next critical decision, whether artificial airway or rigid tracheobronchoscopy was required . Nasotracheal intubation was necessary in 76% of our patients, in one child tracheostomy was performed (5%) . Premature extubation necessitating reintubation occurred in 33% of the children suffering from BT . In these patients, our method of advancing a flexible endoscope for tracheoscopy through the respiration tube failed because of a low tube diameter . Another remarkable finding was the high mortality in AE (14%) . Affected children were admitted in poor post-hypoxia conditions following outdoor cardiorespiratory arrest . CONCLUSION . In the analysis of the clinical course we found three decisive turning points in managing the disorder; First, the confirmation of the correct admission diagnosis; second, the decision, as to whether an artificial airway should be established; and third, the proper time of extubation . The most decisive factor in decreasing mortality seems to be timely, appropriate presentation at referral centers if AE or BT is suspected . Clinically, progressive management of childhood stridor requires close cooperation between the Pediatric, Anesthesiologic, and ENT Departments . Fiberoptic endoscopy as a guide to current airway management is a major step forward and should be a part of every established protocol. World J Surg, 1996 May, 20(4), 411 - 7 Role of the gut in multiple organ failure: bacterial translocation and permeability changes; Swank GM et al.; It is clear that increased gut permeability and bacterial translocation play a role in multiple organ failure (MOF) . Failure of the gut barrier remains central to the hypothesis that toxins escaping from the gut lumen contribute to activation of the host's immune inflammatory defense mechanisms, subsequently leading to the autointoxication and tissue destruction seen in the septic response characteristic of MOF . However, the role of the gut is more than that of a sieve, which simply allows passage of bacteria and endotoxin from the gut lumen to the portal or systemic circulation . It appears, in addition, that the translocation of bacteria and endotoxin may lead to local activation of the immune inflammatory system and the local production of cytokines and other immune inflammatory mediators . These intestinally derived mediators may then exacerbate the systemic inflammatory response and potentially lead to a further increase in gut permeability . A vicious cycle of increased intestinal permeability, leading to toxic mediator release, resulting in a further increase in gut permeability is generated . Additionally, the systemic and local inflammatory cells that become activated in the gut contribute to the systemic response characteristic of the sepsis syndrome and MOF . Thus even if the immune inflammatory system, rather than the gut, is the "motor of" MOF, the gut remains one of the major pistons that turns the motor. Hepatology, 1996 May, 23(5), 1200 - 7 B