Sci Prog, 2003, 86(Pt 1-2), 115 - 37 Lethal effects of heat on bacterial physiology and structure; Russell AD; High temperatures have profound effects on the structural and physiological properties of sporulating and non-sporulating bacteria, with membranes, RNA, DNA, ribosomes, protein and enzymes all affected . Nevertheless, it is apparent that no one single event is responsible for cell death . The induction of intracellular heat-shock proteins and the activation of extracellular alarmones in vegetative cells exposed to mildly lethal temperatures are important cell responses . In bacterial spores, several factors contribute to the overall resistance to moist (wet) and dry heat; the latter, but not the former, induces mutations . Heat resistance develops during sporulation, when spore-specific heat-shock proteins are also produced . Heat sensitivity is regained during germination of spores. Sci Prog, 2003, 86(Pt 1-2), 9 - 75 Bacterial cold shock responses; Weber MH et al.; As a measure for molecular motion, temperature is one of the most important environmental factors for life as it directly influences structural and hence functional properties of cellular components . After a sudden increase in ambient temperature, which is termed heat shock, bacteria respond by expressing a specific set of genes whose protein products are designed to mainly cope with heat-induced alterations of protein conformation . This heat shock response comprises the expression of protein chaperones and proteases, and is under central control of an alternative sigma factor (sigma 32) which acts as a master regulator that specifically directs RNA polymerase to transcribe from the heat shock promotors . In a similar manner, bacteria express a well-defined set of proteins after a rapid decrease in temperature, which is termed cold shock . This protein set, however, is different from that expressed under heat shock conditions and predominantly comprises proteins such as helicases, nucleases, and ribosome-associated components that directly or indirectly interact with the biological information molecules DNA and RNA . Interestingly, in contrast to the heat shock response, to date no cold-specific sigma factor has been identified . Rather, it appears that the cold shock response is organized as a complex stimulon in which post-transcriptional events play an important role . In this review, we present a summary of research results that have been acquired in recent years by examinations of bacterial cold shock responses . Important processes such as cold signal perception, membrane adaptation, and the modification of the translation apparatus are discussed together with many other cold-relevant aspects of bacterial physiology and first attempts are made to dissect the cold shock stimulon into less complex regulatory subunits . Special emphasis is placed on findings concerning the nucleic acid-binding cold shock proteins which play a fundamental role not only during cold shock adaptation but also under optimal growth conditions. J Bacteriol, 2003 Jul, 185(14), 4226 - 32 A bacterial TrwC relaxase domain contains a thermally stable alpha-helical core; Arrondo JL et al.; The TrwC protein is the relaxase-helicase responsible for the initiation and termination reactions of DNA processing during plasmid R388 conjugation . The TrwC-N275 fragment comprises the 275-amino-acid N-terminal domain of the protein that contains the DNA cleavage and strand transfer activities (the relaxase domain) . It can be easily purified by keeping a cell lysate at 90 degrees C for 10 min . Infrared spectroscopy shows that this domain has a predominantly alpha/beta structure with some amount of unordered structure . Fast heating and cooling does not change the secondary structure, whereas slow heating produces two bands in the infrared spectrum characteristic of protein aggregation . The denaturation temperature is increased in the protein after the fast-heating thermal shock . Two-dimensional infrared correlation spectroscopy shows that thermal unfolding is a very cooperative two-state process without any appreciable steps prior to aggregation . After aggregation, the alpha-helix percentage is not altered and alpha-helix signal does not show in the correlation maps, meaning that the helices are not affected by heating . The results indicate that the domain has an alpha-helix core resistant to temperature and responsible for folding after fast heating and an outer layer of beta-sheet and unordered structure that aggregates under slow heating . The combination of a compact core and a flexible outer layer could be related to the structural requirements of DNA-protein binding. Mol Genet Genomics, 2003 Aug, 269(5), 685 - 91 Epub 2003 Jun 27. Construction and application of a bacterial artificial chromosome (BAC) library of Prunus armeniaca L . for the identification of clones linked to the self-incompatibility locus; Vilanova S et al.; To facilitate gene discovery in the Rosaceae, a bacterial artificial chromosome (BAC) library was constructed using high-molecular-weight (HMW) DNA from apricot leaves (Prunus armeniaca L.) . The library contains 101,376 clones (264, 384-well plates) with an average insert size of 64 kb, equivalent to 22-fold genome coverage . In the first application of this library, high-density filters were screened for self-incompatibility genes using apricot DNA probes . Eight positive BAC clones were detected and fingerprinted to determine clone relationships and assemble contigs . These results demonstrate the suitability of this library for gene identification and physical mapping of the apricot genome. Nat Med, 2003 Jul, 9(7), 831 - 5 CpG motifs: the active ingredient in bacterial extracts? Krieg AM. The use of bacteria and bacterial extracts for immunotherapy has a checkered past . Recent developments in immunology reveal that these nonspecific immune activators actually work by triggering specific receptors that are expressed by subsets of immune cells . Identification of these receptors and the molecular signaling pathways that they activate has enabled a new era of specific targeted immunotherapy using chemically synthesized mimics of pathogen molecules. Biomaterials, 2003 Oct, 24(22), 3877 - 84 Binding and orientation of fibronectin to silanated glass surfaces using immobilized bacterial adhesin-related peptides; Klueh U et al.; Previously, we have demonstrated the suitability of bacterial adhesin-related peptides, directly immobilized on polystyrene surfaces, to bind and orient fibronectin (FN) . For these studies a method to bind the large protein FN in a desired orientation on a solid substratum was developed which utilizes a bacterial adhesin-related peptide (designated BRP-A), which is known to bind specifically to the NH3-terminus end of FN . Glass substrata was first coated with an amine-terminated silane, followed by streptavidin (SA), which was used as an intermediate tether to bind the biotinylated bacterial adhesin-related peptide . The BRP-A peptide, used for these studies was synthesized with a terminal biotin to assure irreversible coupling of the BRP-A to the streptavidin . The biotinylated BRP-A was next immobilized on the SA-silanated glass surfaces . 125I-FN was used to quantify the amount of FN binding to the (BRP-A):SA-silanated glass surface . Monoclonal antibodies, which react with specific epitopes at either the NH3- or -COOH-termini of FN, were used to quantify the binding and orientation of FN . The results of these studies indicated: (1) FN bound to the BRP-A:SA-silanated glass surface; and (2) the bound FN was oriented such that NH2-terminal region of FN was bound towards the glass surface and the COOH-terminus was oriented away from the glass surface . These studies demonstrate that small peptides can be used to specifically bind and orient large proteins such as FN on the surfaces. Clin Nephrol, 2003 Jun, 59(6), 447 - 54 Differences in the permeability of high-flux dialyzer membranes for bacterial pyrogens; Schindler R et al.; AIMS: The increasing use of high-flux membranes for hemodialysis has raised concerns that patients dialyzed with these membranes may be at higher risk of being exposed to cytokine-inducing bacterial substances in the dialysate than patients dialyzed with low-flux membranes . We investigated the permeability of various high-flux membranes for both purified E . coli lipopolysaccharide (LPS) as well as for LPS derived from Stenotrophomonas (Sten.) maltophilia . MATERIALS AND METHODS: An in vitro dialysis circuit with saline in the blood compartment of 3 dialyzers containing different membranes (polysulfone, helixone and Diapes) was employed . The dialysate was challenged with increasing doses of sterile filtrates derived from Sten . maltophilia cultures or with purified LPS from E . coli . Samples from the blood compartment were tested for cytokine induction (IL-1beta, IL-6 and TNF) in mononuclear cells as well as for LPS by limulus amebocyte lysate test (LAL) . RESULTS: IL-6 induction above sterile controls (< 0.02 ng/ml IL-6) was observed by samples from the blood side of DIAPES dialyzers (1.2 +/- 0.7 ng/ml IL-6) after challenging the dialysate with 4.1 +/- 3.6 U/ml E . coli LPS (9.9 +/- 4.5 ng/ml IL-6) . In contrast, at the same challenge dose no significant IL-6 induction above sterile controls was observed by blood side samples of polysulfone (0.15 +/- 0.07 ng/ml) and helixone (0.09 +/- 0.05 ng/ml) dialyzers . Increasing the amount of E . coli LPS in the dialysate further augmented IL-6 induction by blood side samples of Diapes but not of polysulfone and helixone dialyzers . Similar results were obtained for IL-1beta and TNF . After challenging the dialysate with E . coli LPS as well as with cultures of Sten . maltophilia, significantly more LAL reactivity was observed in the blood compartment of Diapes compared to polysulfone and helixone . CONCLUSIONS: There are considerable differences between high-flux membranes regarding their permeability for cytokine-inducing substances from E . coli as well as for LPS derived from E . coli and Sten . maltophilia . Dialyzers that leak CIS under aqueous conditions in vivo should not be used unless the dialysate has passed through an ultrafilter. Acta Crystallogr D Biol Crystallogr, 2003 Jul, 59(Pt 7), 1291 - 3 Epub 2003 Jun 27. Crystallization and preliminary X-ray data investigation of the bacterial enterocin A immunity protein at 1.65 A resolution; Dalhus B et al.; Crystals of the bacterial enterocin A immunity protein have been prepared by the hanging-drop vapour-diffusion technique at 293 K . The crystals diffract to better than 1.7 A resolution and X-ray diffraction data to 1.65 A have been collected at 110 K using synchrotron radiation . The enterocin A immunity protein crystals belong to the monoclinic crystal system, with unit-cell parameters a = 116.32, b = 42.35, c = 66.17 A, beta = 111.3 degrees . The symmetry and systematic absences in the diffraction pattern are consistent with space group C2 . The presence of two molecules in the asymmetric unit with a molecular weight of approximately 12.2 kDa gives a crystal volume per protein mass (V(M)) of approximately 3.1 A(3) Da(-1) and a solvent content of approximately 60% by volume. Thorax, 2003 Jul, 58(7), 605 - 12 Enhancement of acute lung injury related to bacterial endotoxin by components of diesel exhaust particles; Yanagisawa R et al.; BACKGROUND: Diesel exhaust particles (DEP) synergistically aggravate acute lung injury related to lipopolysaccharide (LPS) in mice, but the components in DEP responsible for this have not been identified . A study was undertaken to examine the effects of the organic chemicals (DEP-OC) and residual carbonaceous nuclei (washed DEP) derived from DEP on LPS related lung injury . METHODS: ICR mice were divided into experimental groups and vehicle, LPS, washed DEP, DEP-OC, washed DEP+LPS, and DEP-OC+LPS were administered intratracheally . The cellular profile of the bronchoalveolar lavage (BAL) fluid, pulmonary oedema, lung histology, and expression of proinflammatory molecules and Toll-like receptors in the lung were evaluated . RESULTS: Both DEP-OC and washed DEP enhanced the infiltration of neutrophils into BAL fluid in the presence of LPS . Washed DEP combined with LPS synergistically exacerbated pulmonary oedema and induced alveolar haemorrhage, which was concomitant with the enhanced lung expression of interleukin-1beta, macrophage inflammatory protein-1alpha, macrophage chemoattractant protein-1, and keratinocyte chemoattractant, whereas DEP-OC combined with LPS did not . Gene expression of Toll-like receptors 2 and 4 was increased by combined treatment with washed DEP and LPS . The enhancement effects of washed DEP on LPS related changes were comparable to those of whole DEP . CONCLUSIONS: These results suggest that the residual carbonaceous nuclei of DEP rather than the extracted organic chemicals predominantly contribute to the aggravation of LPS related lung injury . This may be mediated through the expression of proinflammatory cytokines, chemokines, and Toll-like receptors. FASEB J, 2003 Jul, 17(10), 1319 - 21 Bacterial DNA evokes epithelial IL-8 production by a MAPK-dependent, NF-kappaB-independent pathway; Akhtar M et al.; Recognition of bacterial products by the innate immune system is dependent on pattern-recognition receptors: toll-like receptor 9 (TLR-9) in the case of bacterial DNA . We hypothesized that bacterial DNA can directly affect enteric epithelial cells . RT-PCR revealed constitutive TLR-9 mRNA expression in three human colonic epithelial cell lines (T84, HT-29, Caco-2) and THP-1 monocytes . Epithelial cells, in six-well culture plates or on filter supports, were exposed to E . coli DNA (1-50 microg/ml), synthetic CpG-rich oligonucleotides, or calf thymus DNA for 6-48 h . Exposure to E . coli DNA resulted in an increase in IL-8 mRNA, and a time- and dose-dependent increase in IL-8 secretion . Also, CpG oligonucleotides induced epithelial IL-8 production, whereas calf thymus DNA did not . Exposure to E . coli DNA resulted in phosphorylation of ERK 1/2 MAPK and inhibitors of ERK activity (PD98059, UO126) significantly reduced the evoked IL-8 production . In contrast, inhibitors of NFkappaB activity (PDTC, SN50) did not block E . coli DNA-induced IL-8 production . Electrophoretic mobility shift assays revealed that E . coli DNA stimulated epithelial AP-1 but not NFkappaB activation . The barrier (i.e., transepithelial resistance) and ion transport parameters of epithelial monolayers (assessed in Ussing chambers) were unaltered following E . coli DNA exposure . Thus model gut epithelia express TLR-9 mRNA and, while maintaining their barrier function, can respond to E . coli DNA by increased IL-8 production. FEMS Immunol Med Microbiol, 2003 Jul 15, 37(2-3), 129 - 34 Progress toward the development of a bacterial vaccine vector that induces high-titer long-lived broadly neutralizing antibodies against HIV-1; Fouts TR et al.; Conformationally constrained HIV-1 Env and gp120 immunogens induce broadly cross-reactive neutralizing antibodies . Thus, it is now feasible to rationally design an HIV-1 vaccine that affords protection through humoral mechanisms . This paper reviews our progress toward the development of an oral bacterial vaccine vector that is capable of delivering an HIV-1 DNA vaccine to host lymphoid tissues and inducing broadly neutralizing antibodies to HIV-1 in the mucosal and systemic immune compartments. FEMS Microbiol Rev, 2003 Jun, 27(2-3), 215 - 37 Bacterial iron homeostasis; Andrews SC et al.; Iron is essential to virtually all organisms, but poses problems of toxicity and poor solubility . Bacteria have evolved various mechanisms to counter the problems imposed by their iron dependence, allowing them to achieve effective iron homeostasis under a range of iron regimes . Highly efficient iron acquisition systems are used to scavenge iron from the environment under iron-restricted conditions . In many cases, this involves the secretion and internalisation of extracellular ferric chelators called siderophores . Ferrous iron can also be directly imported by the G protein-like transporter, FeoB . For pathogens, host-iron complexes (transferrin, lactoferrin, haem, haemoglobin) are directly used as iron sources . Bacterial iron storage proteins (ferritin, bacterioferritin) provide intracellular iron reserves for use when external supplies are restricted, and iron detoxification proteins (Dps) are employed to protect the chromosome from iron-induced free radical damage . There is evidence that bacteria control their iron requirements in response to iron availability by down-regulating the expression of iron proteins during iron-restricted growth . And finally, the expression of the iron homeostatic machinery is subject to iron-dependent global control ensuring that iron acquisition, storage and consumption are geared to iron availability and that intracellular levels of free iron do not reach toxic levels. Hepatogastroenterology, 2003 May-Jun, 50(51), 680 - 3 Octreotide inhibits hepatic fibrosis, bile duct proliferation and bacterial translocation in obstructive jaundice; Turkcapar N et al.; BACKGROUND/AIMS: The protective effect of octreotide on bacterial translocation, bile duct epithelial proliferation and hepatic fibrosis was studied in an experimental obstructive jaundice model . METHODOLOGY: Forty-five healthy Wistar albino rats were randomly divided into three groups . Group I (n = 15): Median laparotomy and common bile duct manipulation performed (Sham group) . Group II (n = 15): Laparotomy and common bile duct ligation performed . Group III (n = 15): After laparotomy and common bile duct ligation octreotide (Sandostatin, sandoz) was given . Simultaneously group I and II received 3 cc 0.9% NaCl and group III received 20 micrograms/kg/daily octreotide subcutaneously every 8 hours during 9 days . Two days after the procedure all rats were opened under ether anesthesia and sterile conditions . Group I had simple laparotomy but group II and III also had common bile duct ligation by 5/0 prolene . Seven days after the surgery (9th day after treatment) all rats underwent laparotomy and tests for bacterial translocation, liver biochemical tests and histopathologic analysis of liver and small bowel were carried out . RESULTS: In group II cecal population levels of bacteria were significantly higher than group I and group III (p < 0.05) . In group II there was also statistically significant bacterial translocation to the mesenteric lymph nodes . Pathological changes were found in terminal ileum samples in group II which seemed to improve in group III . Hepatocyte function was preserved with octreotide treatment which also significantly decreased bile duct proliferation and periportal fibrosis in response to biliary obstruction . CONCLUSIONS: This experimental study showed that octreotide is effective in preventing bacterial translocation, bile duct proliferation and hepatic fibrosis in obstructive jaundice. Proc Natl Acad Sci U S A, 2003 Jul 8, 100(14), 8223 - 8 Epub 2003 Jun 25. Quantitative modeling of sensitivity in bacterial chemotaxis: the role of coupling among different chemoreceptor species; Mello BA et al.; We propose a general theoretical framework for modeling receptor sensitivity in bacterial chemotaxis, taking into account receptor interactions, including those among different receptor species . We show that our model can quantitatively explain the recent in vivo measurements of receptor sensitivity at different ligand concentrations for both mutant and wild-type strains . For mutant strains, our model can fit the experimental data exactly . For the wild-type cell, our model is capable of achieving high gain while having modest values of Hill coefficient for the response curves . Furthermore, the high sensitivity of the wild-type cell in our model is maintained for a wide range of ambient ligand concentrations, facilitated by near-perfect adaptation and dependence of ligand binding on receptor activity . Our study reveals the importance of coupling among different chemoreceptor species, in particular strong interactions between the aspartate (Tar) and serine (Tsr) receptors, which is crucial in explaining both the mutant and wild-type data . Predictions for the sensitivity of other mutant strains and possible improvements of our model for the wild-type cell are also discussed. Cytokine, 2003 Mar 21, 21(6), 295 - 302 Excessive increase of serum interleukin 6 jeopardizes host defense against multi-bacterial infection; Soda K et al.; Serum interleukin 6 (IL-6) is elevated among patients who have undergone surgery, trauma, and thermal injury . It is well known that the greater the increase of serum IL-6, the higher the incidence of post-injury morbidity and mortality is . However, it has not been determined whether the physiological effects of IL-6 increase the rate of morbidity and mortality or if IL-6 is just a bystander that only indicates the severity of the injury . To elucidate this, we planned to investigate the effect of IL-6 on a multi-bacterial infection, one of the most frequent post-injury complications . CDF1 male mice were administered recombinant human IL-6 (hIL-6) continuously at a dose of 0, 1, or 10 microg/day . The mice then underwent cecal ligation without puncture that induced slow multi-bacterial infection . The survival rate of mice receiving 10 microg/day of hIL-6 was significantly lower (38.5%) than the rate of those receiving 0 (83.3%) or 1 (92.3%) microg/day of hIL-6 . The result of this study showed that only excessive increases in serum IL-6, to levels that were observed among patients who underwent severe injury or extensive surgery with high incidence of post-injury infection, jeopardize the host's defense against bacterial infection. Trends Microbiol, 2003 Jun, 11(6), 280 - 5 RNA-mediated control of virulence gene expression in bacterial pathogens; Johansson J et al.; Until recently, gene expression was thought to be controlled mainly at the level of transcription initiation by repressor or activator proteins . In some cases, transcription elongation is controlled by a so-called attenuation mechanism that involves alternative base-pairing between different regions of an mRNA transcript . Recent data reveal that other mechanisms can regulate gene expression and involve RNAs that might act as antisense RNAs, sequestering molecules, or thermosensors . This review focuses on recent studies in bacterial pathogens in which a growing list of examples show that RNA can control virulence gene expression. Brain, 2003 Aug, 126(Pt 8), 1873 - 82 Epub 2003 Jun 23. Lack of IL-6 augments inflammatory response but decreases vascular permeability in bacterial meningitis; Paul R et al.; Interleukin (IL)-6 is a multifunctional cytokine with diverse actions and has been implicated in the pathophysiology of many neurological and inflammatory disorders . In this study, we investigated the role of IL-6 in pneumococcal meningitis . Cerebral infection in wild-type (WT) mice caused an increase in vascular permeability and intracranial pressure (ICP), which were significantly reduced in IL-6-/- mice . In contrast, meningitis in IL-6-/- mice was associated with a significant increase in CSF white blood cell count compared with infected WT mice, indicating an enhanced inflammatory response . Analysis of mRNA expression in the brain showed an increase in tumour necrosis factor (TNF)-alpha, IL-1beta, and macrophage inflammatory protein 2 (MIP-2) levels, but decreased expression of granulocyte-macrophage colony-stimulating factor in infected IL-6-/- mice compared with infected WT controls . Similar results were obtained when rats challenged with pneumococci were systemically treated with neutralizing anti-IL-6 antibodies, resulting in an increased pleocytosis but at the same time a reduction of vascular permeability, brain oedema formation, and ICP, which was not accompanied by a downregulation of matrix metalloproteinases . Our data indicate that IL-6 plays an important anti-inflammatory role in bacterial meningitis by reducing leukocyte infiltration but contributes to the rise in intracranial pressure by increasing blood-brain barrier (BBB) permeability . These findings suggest that the migration of leukocytes across the BBB and the increase in vascular permeability are two independent processes during bacterial meningitis. Infect Immun, 2003 Jul, 71(7), 3766 - 74 Regulation of pulmonary and systemic bacterial lipopolysaccharide responses in transgenic mice expressing human elafin; Sallenave JM et al.; The control of lung inflammation is of paramount importance in a variety of acute pathologies, such as pneumonia, the acute respiratory distress syndrome, and sepsis . It is becoming increasingly apparent that local innate immune responses in the lung are negatively influenced by systemic inflammation . This is thought to be due to a local deficit in cytokine responses by alveolar macrophages and neutrophils following systemic bacterial infection and the development of a septic response . Recently, using an adenovirus-based strategy which overexpresses the human elastase inhibitor elafin locally in the lung, we showed that elafin is able to prime lung innate immune responses . In this study, we generated a novel transgenic mouse strain expressing human elafin and studied its response to bacterial lipopolysaccharide (LPS) when the LPS was administered locally in the lungs and systemically . When LPS was delivered to the lungs, we found that mice expressing elafin had lower serum-to-bronchoalveolar lavage ratios of proinflammatory cytokines, including tumor necrosis factor alpha (TNF-alpha), macrophage inflammatory protein 2, and monocyte chemoattractant protein 1, than wild-type mice . There was a concomitant increase in inflammatory cell influx, showing that there was potential priming of innate responses in the lungs . When LPS was given systemically, the mice expressing elafin had reduced levels of serum TNF-alpha compared to the levels in wild-type mice . These results indicate that elafin may have a dual function, promoting up-regulation of local lung innate immunity while simultaneously down-regulating potentially unwanted systemic inflammatory responses in the circulation. Arch Dis Child, 2003 Jul, 88(7), 615 - 20 Diagnosis and treatment of bacterial meningitis; El Bashir H et al.; This review focuses on recent advances of topical interest regarding the diagnosis and treatment of common causes of bacterial meningitis occurring in children beyond the neonatal period . Tuberculous meningitis is beyond the scope of this review. Eur J Obstet Gynecol Reprod Biol, 2003 Jul 1, 109(1), 67 - 71 Efficacy of the combination of 2 g oral tinidazole and acidic buffering vaginal gel in comparison with vaginal clindamycin alone in bacterial vaginosis: a randomized, investigator-blinded, controlled trial; Milani M et al.; OBJECTIVE: To evaluate the efficacy of tinidazole (T) (Trimonase, Mipharm, Italy) and an acidic vaginal gel (Miphil) (M) in comparison with vaginal clindamycin (CL) (Cleocin Pharmacia Upjohn) in BV . DESIGN: A multicentre, randomised, investigator-blinded, controlled trial . POPULATION AND METHODS: 64 women with BV were enrolled . Thirty-two were allocated to receive oral T 2g, single dose, and 32 were assigned to CL 2% for 7 consecutive days . After week 1, T group were treated with an acidic vaginal gel, 2g every 3 days, for additional 3 weeks, whereas CL group did not received any additional treatment . Patients were evaluated at week 1 and 4 . Vaginal pH, the BV-blue test (Gryphus Diagnostics, USA) and the whiff test were performed at baseline and at week 4 . MAIN OUTCOMES MEASURES: Clinical cure rate; normalisation of vaginal pH (pH<4.5); and laboratory cure rate (defined as a clinical cure rate and a negative results of BV-blue and whiff test) . RESULTS: At baseline, vaginal pH values were (mean+/-S.D.) 5.4+/-0.7 and 5.3+/-0.5 in T and CL groups, respectively . Six patients (2 in T group and 4 in CL group) withdrew from the study due to side effects . At week 1, the clinical cure rates were 84% in both T and CL treated group (P=N.S.) . At week 4, clinical cure rates were 94% in T+M group and 77% in CL group (P=N.S.) . The laboratory cure rates were 81% in T+M group and 59% in CL group (P<0.04) . Vaginal pH normalisation (i.e . pH <4.5) was achieved in 78% and in 38% of T+M and CL groups, respectively (P<0.0007) . CONCLUSIONS: In the short term, 2g single oral dose tinidazole was at least as effective as 7-day of vaginal clindamycin . The sequential treatment of tinidazole and acidic vaginal gel was superior to vaginal clindamycin in lowering vaginal pH and achieving a higher laboratory tests normalization rate at 1-month follow-up. Ultrason Sonochem, 2003 Jul, 10(4-5), 231 - 4 The development and evaluation of electrolysis in conjunction with power ultrasound for the disinfection of bacterial suspensions; Joyce E et al.; There is an increasing incidence in health problems related to environmental issues that originate from inadequate treatment of potable waters . This has compelled scientists and engineers to engage in innovative technologies to achieve a maximum disinfection at affordable costs . Some species of bacteria produce colonies and spores that can agglomerate in spherical clusters and thus protect organisms on the inside of the cluster against biocidal attack . Flocs of fine particles (e.g., clay) can entrap bacteria and this can also protect them against the biocides . Other bacteria have the ability to mutate, thus building up resistance to conventional biocides (e.g., chlorine).Ultrasound has been shown to be effective in improving the effectiveness of biocides such as chlorine . The aim of this present study was to investigate the effect of electrolysis and power ultrasound as a disinfection treatment and to provide a greater knowledge of the fundamentals of disinfection through the production of hypochlorite in situ from saline solution via electrolysis . The electrode materials investigated were, carbon (felt and graphite), copper and stainless steel rods . The results show that sonication appears to amplify the effect of electrolysis . A combination of both treatments is significantly better than sonication or electrolysis alone. Am J Gastroenterol, 2003 Jun, 98(6), 1362 - 70 Small intestinal motility disturbances and bacterial overgrowth in patients with liver cirrhosis and portal hypertension; Gunnarsdottir SA et al.; OBJECTIVE: Altered small bowel motility and a high prevalence of small intestinal bacterial overgrowth (SIBO) has been observed in patients with liver cirrhosis . Our aim was to explore the relationship between motility abnormalities, portal hypertension, and SIBO . METHODS: Twenty-four patients with liver cirrhosis were included . Twelve had portal hypertension (PH) and 12 had liver cirrhosis (LC) alone . Child-Pugh score was the same in the groups . Antroduodenojejunal pressure recordings were performed, and noninvasive variceal pressure measurements were undertaken . Thirty-two healthy volunteers served as a reference group . Bacterial cultures were obtained from jejunal aspirates . RESULTS: The PH group had a higher proportion of individual pressure waves that were retrograde in the proximal duodenum during phase II (52% vs 13% vs 8% of propagated contractions; p < 0.001) as well as postprandially (49% vs 18% vs 13%; p < 0.01) compared with LC and controls, respectively . Long clusters were more common in PH than in controls (9.1 +/- 2.1 vs 4.9 +/- 0.8; p < 0.05), and a higher motility index in phase III in the proximal and distal duodenum was seen in the PH as compared with the other groups . The mean variceal pressure was 21 +/- 1 mm Hg . Motor abnormalities were not correlated to the level of variceal pressure . Thirty-three percent of the patients in the PH group but none in the LC group had SIBO . CONCLUSIONS: Abnormal small bowel motility and SIBO is common in patients with liver cirrhosis with concomitant portal hypertension . Portal hypertension per se might be significantly related to small bowel abnormalities observed in patients with liver cirrhosis. Chin J Physiol, 2002 Jun 30, 45(2), 69 - 74 Enhancement of CREBSerine-133 phosphorylation through nitric oxide-mediated signaling induced by bacterial lipopolysaccharide in vascular smooth muscle cells from rats; Yang SN et al.; Nitric oxide (NO) induced by bacterial lipopolysaccharide (LPS) plays a critical role in various patho-physiological implications, such as atherosclerosis, vasculitis and septic shock . In addition, cAMP-responsive element binding protein (CREB), an important transcription factor for cell differentiation, has been shown to be involved in atherosclerogenesis in VSMCs . Here we investigated the possibility whether LPS-induced NO signaling led to phosphorylation of cAMP-responsive element binding protein on Serine-133 (CREBSer-133) in cultured vascular smooth muscle cells (VSMCs) from rats . Addition of LPS (1-10 microg/ml) for 48 hours increased not only the production NO, but also the phosphorylation of CREBSer-133 . The use of NOS inhibitor (100-500 microM L-NAME) blocked the magnitudes of both LPS-induced NO production and CREBSer-133 phosphorylation . In addition, either a guanylyl cyclase (GC) inhibitor (30 microM ODQ) or a cGMP-dependent protein kinase (PKG) inhibitor (20 microM (Rp)-8-pCPT-cGMPs) significantly attenuated the magnitudes of LPS-induced CREBSer-133 phosphorylation, suggesting the involvement of NO-GC-PKG signaling . Thus, the present study suggests that NO-mediated signaling activated by bacterial LPS, at least in part, enhance CREBSer-133 phosphorylation in cultured VSMCs . The findings here may provide not only signaling pathway involved in VSMC differentiation during inflammatory response, but also new insight into possible therapeutic intervention. Philos Transact A Math Phys Eng Sci, 2003 Jun 15, 361(1807), 1283 - 312 Bacterial self-organization: co-enhancement of complexification and adaptability in a dynamic environment; Ben-Jacob E; During colonial development, bacteria generate a wealth of patterns, some of which are reminiscent of those occurring in abiotic systems . They can exhibit rich behaviour, reflecting informative communication capabilities that include exchange of genetic materials and the fact that the colony's building blocks are biotic . Each has internal degrees of freedom, informatic capabilities and freedom to respond by altering itself and others via emission of signals in a self-regulated manner . To unravel the special secrets of bacterial self-organization, we conducted an integrative (experimental and theoretical) study of abiotic and biotic systems . Guided by the notion of general biotic motives and principles, I propose that the informative communication between individuals makes possible the enhancement of the individuals' regulated freedom, while increasing their cooperation . This process is accomplished via cooperative complexification of the colony through self-organization of hierarchical spatio-temporal patterning . The colonial higher complexity provides the degree of plasticity and flexibility required for better colonial adaptability and endurability in a dynamic environment . The biotic system can modify the environment and obtain environmental information for further self-improvement . I reflect on the potential applications of the new understanding on 'engineered self-organization of systems too complex to design' and other issues. Biotechniques, 2003 Jun, 34(6), 1220 - 2, 1224, 1226 passim Bacterial IMPDH gene used for the selection of mammalian cell transfectants; Baccam M et al.; Stable cell transfection is used for the expression of exogenous genes or cDNAs in eukaryotic cells . Selection of these transfectants requires a dominant selectable marker . A variety of such markers has been identified and is currently in use . However, many of these are not suitable for all cell types or require unique conditions . Here we describe a simple and versatile dominant selectable marker that involves bacterial IMP dehydrogenase (IMPDH), an enzyme essential for the replication of mammalian and bacterial cells . Although IMPDH is evolutionarily conserved, the bacterial enzyme is orders of magnitude more resistant to the toxic effect of the drug mycophenolic acid, which is an IMPDH inhibitor . We have demonstrated that transfection of human, monkey or Chinese hamster cell lines with an expression vector containing bacterial IMPDH and mycophenolic acid treatment resulted in the selection of colonies with a strikingly increased resistance to mycophenolic acid toxicity . Analysis of cells derived from these colonies indicated that the acquisition of this resistance was associated with bacterial IMPDH protein expression . As a proof of principle, we showed that mammalian cell transfection with a bicistronic IMPDH/GFP expression vector and mycophenolic acid treatment can be used to successfully select transfectants that express the fluorescent protein . These results indicate that bacterial IMPDH is a practical dominant selectable marker that can be used for the selection of transfectants that express exogenous genes or cDNAs in mammalian cells. J Bacteriol, 2003 Jul, 185(13), 3918 - 25 Archaeal homolog of bacterial type IV prepilin signal peptidases with broad substrate specificity; Albers SV et al.; A large number of secretory proteins in the thermoacidophile Sulfolobus solfataricus are synthesized as a precursor with an unusual leader peptide that resembles bacterial type IV prepilin signal sequences . This set of proteins includes the flagellin subunit but also various solute binding proteins . Here we describe the identification of the S . solfataricus homolog of bacterial type IV prepilin peptidases, termed PibD . PibD is an integral membrane protein that is phylogenetically related to the bacterial enzymes . When heterologously expressed in Escherichia coli, PibD is capable of processing both the flagellin and glucose-binding protein (GlcS) precursors . Site-directed mutagenesis of the GlcS signal peptide shows that the substrate specificity of PibD is consistent with the variations found in proteins with type IV prepilin-like signal sequences of S . solfataricus . We conclude that PibD is responsible for the processing of these secretory proteins in S . solfataricus. Zhonghua Shao Shang Za Zhi, 2003 Apr, 19(2), 89 - 93 {An experimental study on the prevention and treatment of postburn intestinal injury and bacterial translocation by Sijunzi decoction in scalded rats}; Guo L et al.; OBJECTIVE: To explore the effects of traditional Chinese herbal medicine Sijunzi decoction on amelioration of postburn intestinal injury in scalded rats . METHODS: One hundred and eighty Wistar rats were randomly divided into 3 groups, i.e . scald and treatment (T), scald control (S) and normal control (C) groups . The rats in T group were gavaged with the decoction consisting of tangshen, tuckahoe, largehead atractylodes rhizome, glycyrrhizia and rhubarb in a dose of 2 ml twice daily, while the rats in C group were just gavaged with the same amount of distilled water . The rats were sacrificed according to the scheduled postburn observation timepoints . The contents of TNF, NO, MDA and ATPase activity in rat plasma and the intestinal mucosa and the S-IgA content in the intestinal mucus were determined respectively . The changes in histopathology of intestinal mucosa were observed . The samples from internal organ tissue and blood were obtained for bacterial culture . RESULTS: The contents of TNF, NO and MDA in the intestinal mucosa tissue and the rat plasma in scalded rats were lowered significantly by Sijunzi decoction . Furthermore, S-IgA secretion from intestinal mucous cells was maintained by Sijunzi decoction . T cell count was recovered and intestinal mucous barrier injury were lessened, and the bacterial positive rate in the internal organs was decreased . CONCLUSION: Traditional Chinese herbal medicine Sijunzi decoction might be helpful in alleviation of postburn intestinal injury and in the prevention of intestinal bacterial translocation. Eur J Gastroenterol Hepatol, 2003 Jul, 15(7), 739 - 43 Spontaneous bacterial peritonitis in the Czech Republic: prevalence and aetiology; Lata J et al.; The aim of the study was to determine the prevalence and detailed data concerning the incidence of spontaneous bacterial peritonitis in the Czech Republic . Ninety-nine patients with liver cirrhosis and ascites were examined . Spontaneous bacterial peritonitis was diagnosed in 35 patients (35.4%) . It was revealed more often in patients with alcoholic aetiology of cirrhosis whose anamnesis involved sub-febrile or febrile states and the deterioration of ascites . Elevated serum leucocyte counts and increased levels of C-reactive protein can contribute to the diagnosis . A low level of total protein and albumin in ascites predisposes to the increase of this infection . The reduction of the platelet count in a set of patients with spontaneous bacterial peritonitis indicates the influence of portal hypertension in the aetiology of the disease. Biochem Biophys Res Commun, 2003 Jul 4, 306(3), 674 - 9 Down-regulation of Toll-like receptor expression in monocyte-derived Langerhans cell-like cells: implications of low-responsiveness to bacterial components in the epidermal Langerhans cells; Takeuchi J et al.; In the skin, there are unique dendritic cells called Langerhans cells, however, it remains unclear why this particular type of dendritic cell resides in the epidermis . Langerhans cell-like dendritic cells (LCs) can be generated from CD14(+) monocytes in the presence of GM-CSF, IL-4, and TGF-beta1 . We compared LCs with monocyte-derived dendritic cells (DCs) generated from CD14(+) monocytes in the presence of GM-CSF and IL-4 and examined the effect of exposure to two distinct bacterial stimuli via Toll-like receptors (TLRs), such as peptidoglycan (PGN) and lipopolysaccharide (LPS) on LCs and DCs . Although stimulation with both ligands induced a marked up-regulation of CD83 expression on DCs, PGN but not LPS elicited up-regulation of expression CD83 on LCs . Consistent with these results, TLR2 and TLR4 were expressed on DCs, whereas only TLR2 was weakly detected on LCs . These findings suggest the actual feature of epidermal Langerhans cells with low-responsiveness to skin commensals. Hepatol Res, 2003 Jun, 26(2), 114 - 118 Unusual hyperbilirubinemia associated with bacterial pneumonia and acute myeloid leukemia; Harada M et al.; A 77-year-old man with pneumonia associated with acute myeloid leukemia was introduced to the hepatology unit at our hospital for hyperbilirubinemia . He had been suffering from a high fever because of pneumonia . He was icteric and his serum concentrations of total and direct bilirubin were 13.1 and 7.9 mg/dl, respectively . However, the other standard biochemical examinations for hepatic function, such as serum concentrations of aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transpeptidase and alkaline phosphatase were normal except for lactate dehydrogenase . Lactate dehydrogenase isoenzyme analysis revealed that the high concentration was derived from leukemia cells . Ultrasonography of the abdomen revealed no abnormality in the liver or biliary tract . Administration of antibiotics for pneumonia decreased the serum bilirubin concentration, however, he died because of respiratory failure caused by the progression of pneumonia at 33 days after the admission . It was suggested that a disturbance in the bilirubin metabolism without hepatocyte necrosis or mechanical cholestasis might be involved in the pathogenesis of hyperbilirubinemia in patients with infectious diseases. J Biol Chem, 2003 Aug 29, 278(35), 32778 - 83 Epub 2003 Jun 13. The mitochondrial ornithine transporter . Bacterial expression, reconstitution, functional characterization, and tissue distribution of two human isoforms; Fiermonte G et al.; Two isoforms of the human ornithine carrier, ORC1 and ORC2, have been identified by overexpression of the proteins in bacteria and by study of the transport properties of the purified proteins reconstituted into liposomes . Both transport L-isomers of ornithine, lysine, arginine, and citrulline by exchange and by unidirectional mechanisms, and they are inactivated by the same inhibitors . ORC2 has a broader specificity than ORC1, and L- and D-histidine, L-homoarginine, and D-isomers of ornithine, lysine, and ornithine are all substrates . Both proteins are expressed in a wide range of human tissues, but ORC1 is the predominant form . The highest levels of expression of both isoforms are in the liver . Five mutant forms of ORC1 associated with the human disease hyperornithinemia-hyperammonemia-homocitrullinuria were also made . The mutations abolish the transport properties of the protein . In patients with hyperornithinemia-hyperammonemia-homocitrullinuria, isoform ORC2 is unmodified, and its presence compensates partially for defective ORC1. J Negat Results Biomed . 2003 Jun 9;2(1):3. Bacterial adherence to mucosal epithelium in the upper airways has less significance than believed; Ebenfelt A; BACKGROUND: Bacterial adherence to the upper airway epithelium is considered to be an important phenomenon in the pathogenesis of infections . However, the evidence for the hypothesis that bacterial adherence to mucosal epithelial cells has significance for pathogenesis of mucosal infections is based on studies using indirect techniques . We could find no biopsy studies with direct ocular observations of significant numbers of bacteria adhering to upper airway mucosal epithelial cells either in health or during disease . RESULTS: We studied specimens from healthy and infected tonsillar epithelium and specimens from the soft palate epithelium obtained by surgery . The specimens were examined by TEM . In the vast majority of specimens, we found no bacteria adhering to the epithelial cells in the mucosal line regardless of whether the patient was infected or not . Bacteria adhering to shed epithelial cells were seen in higher numbers . Furthermore, as bacteria are small compared to epithelial cells, we calculated the risk of overlooking every adhered bacteria in a section if bacterial adherence was such a significant phenomenon as earlier suggested . We found this risk to be very small . CONCLUSION: We conclude that bacterial adherence to mucosal surface epithelial cells is not a significant phenomenon, either in healthy mucosa in the upper airways or during infection . This is also in line with our earlier results, where we have shown that the site for the infectious process in pharyngotonsillitis is in the secretion on the tonsillar mucosal surface. Plant Physiol, 2003 Jun, 132(2), 949 - 57 Identification and expression analysis of a gene encoding a bacterial-type phosphoenolpyruvate carboxylase from Arabidopsis and rice; Sanchez R et al.; Phosphoenolpyruvate carboxylase (PEPC) is distributed in plants and bacteria but is not found in fungi and animal cells . Important motifs for enzyme activity and structure are conserved in plant and bacterial PEPCs, with the exception of a phosphorylation domain present at the N terminus of all plant PEPCs reported so far, which is absent in the bacterial enzymes . Here, we describe a gene from Arabidopsis, stated as Atppc4, encoding a PEPC, which shows more similarity to Escherichia coli than to plant PEPCs . Interestingly, this enzyme lacks the phosphorylation domain, hence indicating that it is a bacterial-type PEPC . Three additional PEPC genes are present in Arabidopsis, stated as Atppc1, Atppc2, and Atppc3, encoding typical plant-type enzymes . As most plant PEPC genes, Atppc1, Atppc2, and Atppc3 are formed by 10 exons interrupted by nine introns . In contrast, Atppc4 gene has an unusual structure formed by 20 exons . A bacterial-type PEPC gene was also identified in rice (Oryza sativa), stated as Osppc-b, therefore showing the presence of this type of PEPC in monocots . The phylogenetic analysis suggests that both plant-type and bacterial-type PEPCs diverged early during the evolution of plants from a common ancestor, probably the PEPC from gamma-proteobacteria . The diversity of plant-type PEPCs in C3, C4, and Crassulacean acid metabolism plants is indicative of the evolutionary success of the regulation by phosphorylation of this enzyme . Although at a low level, the bacterial-type PEPC genes are expressed in Arabidopsis and rice. Gut, 2003 Jul, 52(7), 927 - 32 Host gastric Lewis expression determines the bacterial density of Helicobacter pylori in babA2 genopositive infection; Sheu BS et al.; BACKGROUND AND AIMS: We tested if host gastric Lewis antigens and the babA2 genotype of Helicobacter pylori correlated with clinicohistological outcome . METHODS: We enrolled 188 dyspeptic patients (45 with duodenal ulcer, 45 with gastric ulcer, and 98 with chronic gastritis) with H pylori infection, proved by culture and gastric histology, reviewed by the updated Sydney system . Gastric expression of Lewis (Le) antigens Le(a), Le(b), Le(x), and Le(y) was determined immunochemically to determine intensity (range 0-3) . The corresponding 188 H pylori isolates were screened for babA2 genotype by polymerase chain reaction . RESULTS: All H pylori isolates had a positive babA2 genotype . We identified Le(a) in 33.5%, Le(b) in 72.9%, Le(x) in 86.2%, and Le(y) in 97.4% of biopsies from these 188 patients . Patients who expressed Le(b) had a higher H pylori density than those who did not express Le(b) (p<0.001) . Among 139 patients who expressed Le(b), H pylori density increased with a higher Le(b) intensity (p<0.05) . Gastric atrophy decreased with Le(b) intensity and thus resulted in lower H pylori density in the antrum (p<0.05) . For the 49 patients without gastric Le(b) expression, H pylori density was positively related with Le(x) and Le(a) expression (p<0.05) . CONCLUSIONS: Taiwanese H pylori isolates are 100% babA2 genopositive . Gastric Le(b) as well as Le(x) intensity may be major determinants of H pylori density . While lacking gastric Le(b) expression, Le(x) and Le(a) were closely related to H pylori colonisation. J Biochem (Tokyo), 2003 May, 133(5), 577 - 81 Production of a functional catalytic antibody ScFv-NusA fusion protein in bacterial cytoplasm; Zheng L et al.; Functional expression of catalytic antibodies in the cytoplasm of E . coli is potentially of great interest in searching for new catalysts by genetic selection . Herein, a catalytic antibody single chain Fv (ScFv) 14D9, which catalyzes a highly enantioselective protonation, was expressed as a NusA fusion protein under the T7 promoter . A functional disulfide-containing ScFv fusion protein was obtained in the oxidizing environment of bacterial cytoplasm . The 14D9 ScFv could not be overexpressed alone without NusA fusion . The highly soluble NusA protein most likely retards aggregate formation of ScFv and indirectly supports correct folding and disulfide bridge formation in the fusion construct ScFv-NusA . The ScFv-NusA fusion product shows highly enantioselective, specific, hapten inhibited catalytic activity comparable to its parent monoclonal antibody, 14D9 . The NusA fusion method might be generally helpful for functional antibody expression in vivo and for the new development of biocatalysts by genetic selection. Biotechnol Bioeng, 2003 Aug 20, 83(4), 474 - 8 Role of water-soluble polysaccharides in bacterial cellulose production; Ishida T et al.; Acetobacter xylinum BPR2001 produces water-insoluble bacterial cellulose (BC) and a water-soluble polysaccharide called acetan in corn steep liquor-fructose medium . Acetobacter xylinum EP1, which is incapable of acetan production was derived by disrupting the aceA gene of BPR2001 . The BC production by EP1 (2.88 g/L) was lower than that by BPR2001 (4.6 g/L) in baffled-flask culture . When purified acetan or agar was added to the medium from the start of cultivation, the BC production by EP1 was enhanced and the final BC yield of EP1 was almost the same as that of BPR2001 . A similar improvement of BC production by EP1 by the addition of agar was also confirmed by cultivation in a 50-L airlift reactor . From these results, the role of acetan in BC production is associated with the increase in the viscosity of the culture medium which may hinder coagulation of BC and cells in the culture, thereby accelerating the growth of BPR2001 and BC production by BPR2001 . Transfus Clin Biol, 2003 Jun, 10(3), 192 - 200 {Transfusion-transmitted bacterial infection: residual risk and perspectives of prevention}; Morel P et al.; Bacterial contamination of blood components represents today the highest infectious risk of blood transfusion, the risk is particularly high when it affects platelet concentrates . The residual risk of transfusion reaction due to bacterial contamination of platelets concentrates remains stable . For all severity 1 case occurs with 25,000 distributed platelets concentrates and 1 death occurs with 200,000 distributed units . In France, efforts have focused on the prevention of contamination during donation--involving measures such as rejecting the first few millilitres of donated blood and improving skin disinfection--and the prevention of bacterial proliferation in platelets concentrates--notably by removing leukocytes and ensuring high-quality storage of donated blood . Improving strategies for reducing the risks of bacterial contamination is one of the priorities of the French National Blood Transfusion Service (l'Etablissement francais du sang-EFS) . There is currently considerable debate about the relative importance of bacterial screening methods and methods for inactivating pathogens present in PC . Automated culture (Biomerieux) and the ScanSystem (Hemosystem) and BDS (Pall) method are the most advanced detection systems available, to our knowledge . In term of pathogen inactivation system for platelets, Intercept (Baxter) is nearing the commercial market . These new prevention have logistic and/or functional consequences that will require close scrutiny methods . A national study group is currently considering the consequences of each of these methods and should give its opinion at the end of the first half of 2003. Bioorg Med Chem Lett, 2003 Jul 7, 13(13), 2231 - 4 Discovery of a potent and selective series of pyrazole bacterial methionyl-tRNA synthetase inhibitors; Finn J et al.; Starting with a micromolar lead identified from high-throughput screening, a series of pyrazoles were discovered with significantly improved potency on bacterial methionyl-tRNA synthetase and selectivity over human methionyl-tRNA synthetase. Infect Genet Evol, 2003 May, 3(1), 47 - 55 Mycoplasma pneumoniae: a reduced-genome intracellular bacterial pathogen; Meseguer MA et al.; Mycoplasma pneumoniae has classically been considered an extracellular (or membrane-associated) organism . Nevertheless, the recently elucidated genomic structure of this pathogen strongly suggest that this organism may have been subjected to the process of reductive genetic evolution which is characteristic of intracellular bacteria . We studied the Mycoplasma pneumoniae RYC15989 strain, recovered from a pericardial biopsy sample from a patient with atypical pneumonia and acute pericarditis . The interaction of this strain with human hepatocytes Hep-G2 and mouse neuroblastoma N2-A cell lines was investigated . Confocal laser scanning microscopy and electronic microscopy evidence is presented of the intracellular location of fluorochrome-labelled Mycoplasma pneumoniae in cell lines infected with the organism in vitro . This finding provides preliminary evidence of cellular invasive capacity of Mycoplasma pneumoniae and casts some new light on the pathogenic potential of Mycoplasma pneumoniae in host infection. J Clin Immunol, 2003 May, 23(3), 202 - 13 Innate immune responses in lupus-prone Palmerston North mice: differential responses to LPS and bacterial DNA/CpG oligonucleotides; Lenert P et al.; Inadequate immune response to infectious danger may contribute to the pathogenesis of systemic autoimmune diseases, e.g., systemic lupus erythematosus . To test this hypothesis, we studied innate responses of prediseased lupus-prone Palmerston North (PN) mice to lipopolysaccharide (LPS), bacterial DNA, and synthetic CpG oligonucleotides . LPS and bacterial DNA/CpG oligodeoxyribonucleotides (ODNs) drove PN splenocytes into the cell cycle and protected B cells against spontaneous apoptosis, as in control lupus-free DBA-1 mice . LPS induced significantly higher IL-6 production in PN than in control splenocytes . In contrast, in PN splenocytes bacterial DNA and CpG ODNs induced approximately four- to sixfold lower IL-12p40 and approximately twofold lower IL-6 secretion than controls . This reduction in cytokine secretion in PN mice was not due to delayed kinetics but was related to significantly higher constitutive and CpG-inducible IL-10 secretion . Neutralizing anti-IL-10 antibodies almost completely restored PN IL-6 and IL-12p40 secretion to DBA-1 levels, whereas exogenous IL-10 inhibited in vitro IL-6 and IL-12p40 production in DBA-1 mice . Importantly, treatment with either IL-10 or anti-IL-10 antibody did not modulate CpG-induced cell cycle entry and apoptosis protection in either strain . In conclusion, lupus-prone PN mice show abnormal innate responses through their pattern-recognition TLR9 receptors, characterized by higher inducible IL-10 and lower IL-12p40 and IL-6 secretion, thus implying that response to infectious danger in PN mice is inappropriate and may be linked to lupus pathogenesis. Am J Clin Oncol, 2003 Jun, 26(3), e54 - 9 Does the addition of glutamine to total parenteral nutrition have beneficial effect on the healing of colon anastomosis and bacterial translocation after preoperative radiotherapy? El-Malt M, Ceelen W, Boterberg T, Claeys G, de Hemptinne B, de Neve W, Pattyn P. Glutamine administration stimulates mucosal growth and preserves the morphology of the intestine . Theoretically, it could improve colonic anastomotic healing after radiotherapy (RT)-induced epithelial damage and mucosal atrophy induced by total parenteral nutrition (TPN) . To investigate this issue, the rectosigmoid colon in male Wistar rats was irradiated to a total dose of 25 Gy . Five days after the end of RT, side-to-side anastomosis was constructed between the irradiated rectosigmoid and the nonirradiated caecum . Postoperatively, animals were divided in three groups: group I, normal diet orally; group II, TPN; group III, TPN enriched with 2% glutamine (Gln-TPN) . All animals decreased in weight during RT and after surgery . Weight regain postoperatively was better in the orally fed animals in comparison with the parenterally fed animals (I vs . II and III; p < 0.01) . Colonic anastomotic bursting pressure (BP) and bursting wall tension (BWT) were significantly less in group II in comparison with groups I and III (II vs . I and III; p < 0.01) . BP and BWT were comparable in groups I and III . No significant differences were found between all the groups in gut bacterial translocation to the blood or to the mesenterical lymph nodes . Conclusively, Gln-TPN can play a role in counteracting the negative effect of food deprivation on the healing of irradiated colonic anastomoses . Postoperative Gln-TPN does not influence gut bacterial translocation in this rat model. Eukaryot Cell, 2003 Jun, 2(3), 646 - 50 Histone-like proteins of the dinoflagellate Crypthecodinium cohnii have homologies to bacterial DNA-binding proteins; Wong JT et al.; The dinoflagellates have very large genomes encoded in permanently condensed and histoneless chromosomes . Sequence alignment identified significant similarity between the dinoflagellate chromosomal histone-like proteins of Crypthecodinium cohnii (HCCs) and the bacterial DNA-binding and the eukaryotic histone H1 proteins . Phylogenetic analysis also supports the origin of the HCCs from histone-like proteins of bacteria. Nutr Rev, 2003 Apr, 61(4), 132 - 5 Small intestinal bacterial overgrowth: a possible risk factor for metabolic bone disease; Anantharaju A et al.; Small intestinal bacterial overgrowth (SIBO) is one of the causes of malabsorption syndromes . The prevalence of metabolic bone disease in patients with SIBO is unknown, but a recent prospective case-control study indicated significant contribution of SIBO to the development of metabolic bone disease . We review this and other reports in the literature and discuss the possible mechanisms causing metabolic bone disease in patients with SIBO. Kyobu Geka, 2003 Jun, 56(6), 501 - 4 {Showing no increase of C-reactive protein and developing non-bacterial median chest wound dehiscence after coronary artery bypass grafting}; Suzuki S et al.; A 72-year-old male underwent coronary artery bypass grafting for 3 vessels disease . After the operation, the peak C-reactive protein (CRP) level was 0.85 mg/dl and CRP levels stayed very low . Forty days after the operation the patient developed a progressive median chest wound dehiscence but the bacterial cultures were negative . In spite of conventional therapies such as debridement of necrotic tissue or irrigation, the wound granulation was underdeveloped . After 6 months an epidermis had developed to cover the wound. Arthritis Rheum, 2003 Jun 15, 49(3), 328 - 34 Bacterial components in the synovial tissue of patients with advanced rheumatoid arthritis or osteoarthritis: analysis with gas chromatography-mass spectrometry and pan-bacterial polymerase chain reaction; Chen T et al.; OBJECTIVE: To study the presence of bacterial components in the synovial tissue (ST) of patients with advanced rheumatoid arthritis (RA) . METHODS: ST was collected during joint surgery from 41 RA patients . Tissue from 39 patients with osteoarthritis (OA), 4 patients with undifferentiated inflammatory arthritis (UA), and 3 cases of accidental deaths served as controls . The pan-bacterial polymerase chain reaction (PCR) with primers for the 23S ribosomal RNA (rRNA) and 16S rRNA genes was used to detect bacterial DNA . In addition, synovial fluid (SF) samples from patients with chlamydial reactive arthritis (ReA) were also examined by the same method . The positive controls, bacterial DNA or ST spiked with different living bacteria, were analyzed alongside clinical samples . Most of the ST samples were also analyzed by gas chromatography-mass spectrometry (GC-MS) for determining the presence of bacteria-derived muramic acid . Strict precautions were followed in the clinics and the laboratory to prevent contamination . RESULTS: In GC-MS analysis, muramic acid was observed in the ST from 4 of 35 RA patients and from 2 of 14 OA patients, but not in ST from 2 patients with UA and 3 cadavers . Bacterial DNA was not detected by either one of the PCR primers used in ST from 42 patients with RA and 39 patients with OA . However, 5 of 15 SF samples from ReA patients were PCR positive . The sensitivity of GC-MS to detect muramic acid was 2 pg/injected amount (227 pg muramic acid/mg ST), and that of the pan-bacterial PCR was 2-20 bacteria colony forming units/reaction . CONCLUSION: These results indicate that a bacterial component, muramic acid, is detectable by GC-MS in ST from a few patients with advanced RA or OA . However, no bacterial DNA was detectable by PCR. Sex Transm Infect, 2003 Jun, 79(3), 254 - 6 National survey of doctors' actions following the diagnosis of a bacterial STD; McCree DH et al.; OBJECTIVES: Little is known about the post-STD diagnosis management practices of community based doctors . The purpose of this study was to describe the reported actions that doctors take after diagnosing gonorrhoea, chlamydia, or syphilis and to determine if these actions differ across the three STDs . METHODS: A random national sample of 7300 doctors (70% response rate) practising in five medical specialties responded to 13 questions related to STD management . Mean differences across STDs were examined using the General Linear Model function of SPSS . RESULTS: Most doctors reported instructing patients to abstain from sex during treatment, to use condoms, and to inform their sexual partners of their exposure after diagnosing gonorrhoea, chlamydia, or syphilis . For syphilis, however, doctors were less likely to treat the patients presumptively and to give them drugs for their partners; and more likely to collect partner information, to follow up with the patient to see if the partner was referred for treatment and to send patient information to the health department . CONCLUSIONS: Doctors' post-STD diagnosis actions were similar for gonorrhoea and chlamydia compared to syphilis . Study findings suggest low levels of STD case reporting and partner follow up by doctors in the sample . Interventions are needed to educate community based doctors about the importance of partner follow up and case reporting in the management of STDs. Microbiol Mol Biol Rev, 2003 Jun, 67(2), 213 - 25, table of contents ADP-glucose pyrophosphorylase, a regulatory enzyme for bacterial glycogen synthesis; Ballicora MA et al.; The accumulation of alpha-1,4-polyglucans is an important strategy to cope with transient starvation conditions in the environment . In bacteria and plants, the synthesis of glycogen and starch occurs by utilizing ADP-glucose as the glucosyl donor for elongation of the alpha-1,4-glucosidic chain . The main regulatory step takes place at the level of ADP-glucose synthesis, a reaction catalyzed by ADP-Glc pyrophosphorylase (PPase) . Most of the ADP-Glc PPases are allosterically regulated by intermediates of the major carbon assimilatory pathway in the organism . Based on specificity for activator and inhibitor, classification of ADP-Glc PPases has been expanded into nine distinctive classes . According to predictions of the secondary structure of the ADP-Glc PPases, they seem to have a folding pattern common to other sugar nucleotide pyrophosphorylases . All the ADP-Glc PPases as well as other sugar nucleotide pyrophosphorylases appear to have evolved from a common ancestor, and later, ADP-Glc PPases developed specific regulatory properties, probably by addition of extra domains . Studies of different domains by construction of chimeric ADP-Glc PPases support this hypothesis . In addition to previous chemical modification experiments, the latest random and site-directed mutagenesis experiments with conserved amino acids revealed residues important for catalysis and regulation. J Bone Joint Surg Br, 2003 May, 85(4), 490 - 4 Body-exhaust suit versus occlusive clothing . A randomised, prospective trial using air and wound bacterial counts; Der Tavitian J et al.; We randomly allocated 50 total knee replacements to scrub teams wearing body-exhaust suits (BES) or Rotecno occlusive clothing . The effectiveness of the clothing was assessed using air andwound bacterial counts . Bacteria were recovered from 62% of wounds (64% BES, 60% Rotecno) . The mean air count was 0.5 CFU/ m3 with BES and 1.0 CFU/m3 with Rotecno (p = 0.014) . The mean wound counts were 14 bacteria/wound with BES and eight bacteria/wound with Rotecno (p = 0.171) . There was no correlation between the air and wound counts (r = -0.011, Spearman's) . The higher air counts suggest that Rotecno occlusive clothing is less effective than BES, but wounds were equally contaminated with both types of clothing suggesting that at very low levels of air contamination the contribution of bacteria to the wound from the air is irrelevant . Even doubling the air counts from 0.5 to 1.0 CFU/m3 had no detectable effect on the wound . This allows a reassessment to be made of other sources of contamination the effect of which would previously have been overwhelmed by contamination from air. Carbohydr Res, 2003 Jun 16, 338(12), 1299 - 308 Synthesis and conformational analysis of the repeating units of bacterial spore peptidoglycan; Keglevic D et al.; Deprotection of the fully blocked disacharide allyl O-(2-amino-4,6-O-benzylidene-3-O-{(R)-1-carboxyethyl}-2-deoxy-beta-D-glucopyranosyl-1',2-lactam)-(1-->4)-2-acetamido-3,6-di-O-benzyl-2-deoxy-beta-D-glucopyranoside by selective de-O-allylation and parallel removal of the benzylidene and O-benzyl groups is described . The resulting beta-muramyl lactam-(1-->4)-GlcNAc disaccharide is characterised as the per-O-acetylated derivative by 1H and 13C NMR spectroscopy and X-ray structure analysis . Conformational analysis about glycosidic bond of repeating units of bacterial spore cortex is based on experimental data and molecular modelling. Proc Natl Acad Sci U S A, 2003 Jun 24, 100(13), 8024 - 9 Epub 2003 Jun 03. Physical interaction between RRS1-R, a protein conferring resistance to bacterial wilt, and PopP2, a type III effector targeted to the plant nucleus; Deslandes L et al.; RRS1-R confers broad-spectrum resistance to several strains of the causal agent of bacterial wilt, Ralstonia solanacearum . Although genetically defined as recessive, this R gene encodes a protein whose structure combines the TIR-NBS-LRR domains found in several R proteins and a WRKY motif characteristic of some plant transcriptional factors and behaves as a dominant gene in transgenic susceptible plants . Here we show that PopP2, a R . solanacearum type III effector, which belongs to the YopJ/AvrRxv protein family, is the avirulence protein recognized by RRS1-R . Furthermore, an interaction between PopP2 and both RRS1-R and RRS1-S, present in the resistant Nd-1 and susceptible Col-5 Arabidopsis thaliana ecotypes, respectively, was detected by using the yeast split-ubiquitin two-hybrid system . This interaction, which required the full-length R protein, was not observed between the RRS1 proteins and PopP1, another member of the YopJ/AvrRxv family present in strain GMI1000 and that confers avirulence in Petunia . We further demonstrate that both the Avr protein and the RRS1 proteins colocalize in the nucleus and that the nuclear localization of the RRS1 proteins are dependent on the presence of PopP2. Biochim Biophys Acta, 2003 Jun 10, 1612(2), 186 - 94 An association of 27- and 40-kDa molecules with glycolipids that bind A-B bacterial enterotoxins to cultured cells; Shimizu T et al.; It is well recognized that the Shiga-like toxins (Stxs) preferentially bind to Gb3 glycolipids and the cholera toxin (CT) and heat-labile enterotoxin (LTp) bind to GM1 gangliosides . After binding to the cell surface, A-B bacterial enterotoxins have to be internalized by endocytosis . The transport of the toxin-glycolipid complex has been documented in several manners but the actual mechanisms are yet to be clarified . We applied a heterobifunctional cross-linker, sulfosuccinimidyl-2-(p-azidosalicylamido)-1,3'-dithiopropionate (SASD), to detect the membrane proteins involved in the binding and the transport of A-B bacterial enterotoxins in cultured cells . Both Stx1 and Stx2 bound to the detergent-insoluble microdomain (DIM) of Vero cells and Caco-2 cells, which were susceptible to the toxin, but neither was bound to insusceptible CHO-K1 cells . Both CT and LTp bound to the DIM of Vero cells, Caco-2 cells, and CHO-K1 cells . In a cross-linking experiment, Stx1 cross-linked only with a 27-kDa molecule, while Stx2, which was more potently toxic than Stx1, cross-linked with 27- and 40-kDa molecules of Vero cells as well as of Caco-2 cells; moreover, no molecules were cross-linked with the insusceptible CHO-K1 cells . LTp was cross-linked only to the 27-kDa molecule of these three cell types but the CT, which was more toxic than LTp, was also cross-linked with 27- and 40-kDa molecules of Vero cells, Caco-2 cells, and CHO-K1 cells . The 27- and the 40-kDa molecules might play a role in the endocytosis and retrograde transport of A-B bacterial enterotoxins. Solid State Nucl Magn Reson, 2003 Jun, 23(4), 198 - 212 Solid-state 13C and 1H spin diffusion NMR analyses of the microfibril structure for bacterial cellulose; Masuda K et al.; To obtain further information about the cause for the rather large splitting of the C4 resonance line into the downfield (C4D) and upfield (C4U) lines in CP/MAS 13C NMR spectra for native cellulose, 13C and 1H spin diffusion measurements have been conducted by using different types of bacterial cellulose samples . In 13C spin diffusion measurements, the C4D resonance line is selectively inverted by the Dante pi pulse sequence and the 13C spin diffusion is allowed to proceed from the C4D carbons to other carbons including the C4U carbons with use of the 13C4-enriched bacterial cellulose sample . The analysis based on the simple spin diffusion theory for the process experimentally observed reveals that the C4U carbons may be located at distances less than about 1 nm from the C4D carbons . In 1H spin diffusion measurements, poly(vinyl alcohol) (PVA) films in which ribbon assemblies of bacterial cellulose are dispersed are employed and the 1H spin diffusion process is examined from the water-swollen PVA continuous phase to the dispersed ribbon assemblies by the 13C detection through the 1H-13C CP technique . As a result, it is found that the C4D and C4U carbons are almost equally subjected to the 1H spin diffusion from the PVA phase, indicating that the C4U carbons are not localized in some limited area, e.g . in the surfacial region, but are distributed in the whole area in the microfibrils . These experimental results suggest that the C4U carbons may exist as structural defects probably due to conformational irregularity associated with disordered hydrogen bonding of the CH(2)OH groups in the microfibrils. Microbes Infect, 2003 Jun, 5(7), 621 - 7 Inducible nitric oxide synthase and control of intracellular bacterial pathogens; Chakravortty D et al.; Inducible nitric oxide synthase (iNOS) has important functions in innate immunity and regulation of immune functions . Here, the role of iNOS in the pathogenesis of various intracellular bacterial infections is discussed . These pathogens have also evolved a broad array of strategies to repair damage by reactive nitrogen intermediates, and to suppress or inhibit functions of iNOS. Mol Microbiol, 2003 Jun, 48(5), 1349 - 55 Oligomerization and activation of the FliI ATPase central to bacterial flagellum assembly; Claret L et al.; FliI is the peripheral membrane ATPase pivotal to the type III protein export mechanism underlying the assembly of the bacterial flagellum . Gel filtration and multiangle light scattering showed that purified soluble native FliI protein was in a monomeric state but, in the presence of ATP, FliI showed a propensity to oligomerize . Electron microscopy revealed that FliI assembles to a ring structure, the yield of which was increased by the presence of a non-hydrolysable ATP analogue . Single particle analysis of the resulting electron micrograph images, to which no symmetry was applied, showed that the FliI ring structure has sixfold symmetry and an external diameter of approximately 10 nm . The oligomeric ring has a central cavity of 2.5-3.0 nm, which is comparable to the known diameter of the flagellar export channel into which export substrates feed . Enzymatic activity of the FliI ATPase showed positive co-operativity, establishing that oligomerization and enzyme activity are coupled . Escherichia coli phospholipids increased enzyme co-operativity, and in vitro cross-linking demonstrated that they promoted FliI multimerization . The data reveal central facets of the structure and action of the flagellar assembly ATPase and, by extension, the homologous ATPases of virulence-related type III export systems. Phys Rev E Stat Nonlin Soft Matter Phys . 2003 May;67(5 Pt 1):051921 . Epub 2003 May 23. Applicability of the Fisher equation to bacterial population dynamics; Kenkre VM et al.; The applicability of the Fisher equation, which combines diffusion with logistic nonlinearity, to population dynamics of bacterial colonies is studied with the help of explicit analytic solutions for the spatial distribution of a stationary bacterial population under a static mask . The mask protects bacteria from ultraviolet light . The solution, which is in terms of Jacobian elliptic functions, is used to provide a practical prescription to extract Fisher equation parameters from observations and to decide on the validity of the Fisher equation. Ann Otol Rhinol Laryngol, 2003 May, 112(5), 461 - 8 Effects of bacterial toxins on air-exposed cultured human respiratory sinus epithelium; Nell MJ et al.; The present study was designed to investigate the effects of the bacterial toxins lipopolysaccharide (LPS) and lipoteichoic acid (LTA) on air-exposed cultured human respiratory sinus epithelium . The morphological changes, proliferation, and differentiation of sphenoid sinus mucosa were examined after incubation with different LPS or LTA concentrations . Air-exposed cultured sinus mucosa differentiated from pseudostratified respiratory epithelium to squamous ciliated epithelium with few goblet cells . High concentrations of bacterial toxins induced a significant increase in mucus production and a decrease in ciliated cells . Ki67 immunostaining showed an increased cell proliferation after incubation with moderate levels of LPS or LTA . High concentrations of bacterial toxins, on the other hand, induced a decreased proliferation . Involucrin expression was clearly altered by incubation with high levels of bacterial toxins, indicating an increased degree of terminal differentiation . These results indicate that the bacterial toxins LPS and LTA both induce comparable dose-dependent morphological changes in sinus epithelium. Ned Tijdschr Tandheelkd, 2003 May, 110(5), 178 - 80 {110th year Nederlands Tijdschrift voor Tandheelkunde . 2 . Root canal treatment, intra-canal disinfectants and bacterial culture: past and present}; Moorer WR et al.; Fifty years ago the Dutch Journal of Dentistry published methods and opinions concerning root canal treatment . Qualitative bacterial culture, inclusion of aggressive disinfectants, as well as antibiotics and widening of the apical constriction were carried out . Nowadays, because of several reasons, these are not clinical practice anymore . Controversy over the clinical consequences of bacterial presence in tubules and in the peri-apical area prevailed in the past and seem to be prevalent once again. Calcif Tissue Int . 2003 Jun 6; {Epub ahead of print} Systemic Effects on Bone Healing of a New Hyaluronic Acid-Like Bacterial Exopolysaccharide; Zanchetta P et al.; Critical Size Defect (CSD) technique was used to evaluate the systemic activities on bone regeneration capacity of a newly discovered hyaluronic acid-like exopolysaccharide synthesized by a bacteria originating from a deep sea hydrothermal vent . Some systemic effects were previously detected on earlier experiments . A 5 mm diameter hole was made on each parietal bone of male rats . The right hole was filled with 0.5 mg of a new bacterial exopolysaccharide referenced HE 800, while the left hole remained free of any treatment . After 21 days, the holes and surrounding tissues were examined by direct examination, X-rays, and histological staining . Using HE 800, bone healing was almost complete after only 21 days in the treated hole and always complete in the control side by some systemic effect . Neovascularization was also observed along with an organized trabecular bone on both sides . No abnormal bone growth or conjunctival abnormalities were noticed . At the end of the experiment, 90.1% (+/-5.2) bone healing (n = 20) was observed on the treated side; conversely, the control side animals demonstrate an amazing healing 100% (+/-0.5) by a systemic effect. Sex Transm Dis, 2003 Jun, 30(6), 483 - 9 Evaluation of a new rapid diagnostic kit (FemExam) for bacterial vaginosis in patients with vaginal discharge syndrome in The Gambia; West B et al.; BACKGROUND: Diagnosis of bacterial vaginosis (BV) in resource-poor primary health care settings is often overlooked; there is a need for a cheap, rapid, objective point-of-care diagnostic test . GOAL: The goal was to determine the prevalence of BV and to evaluate the performance of a new commercial diagnostic test kit in a developing country environment . STUDY DESIGN: Vaginal and cervical swabs were collected from 230 consecutive women attending a genitourinary medicine clinic with reported symptoms of vaginal discharge and/or itching . Etiological testing was carried out . BV was diagnosed on the basis of the Nugent score, the Amsel clinical criteria, and results of FemExam card tests . Card 1 is for pH and amines, and card 2 measures proline iminopeptidase (PIP) activity . RESULTS: BV prevalence was 47.9% according to the Nugent score . When compared with the Nugent score, the Amsel clinical criteria had a sensitivity of 77.9% and specificity of 58.4%, FemExam card 1 had a sensitivity of 71.4% and specificity of 72.8%, FemExam card 2 had a sensitivity of 70% and specificity of 81.0%, and FemExam cards 1 and 2 combined had a sensitivity of 91.0% and specificity of 61.5% . Cost per patient and cost per true case detected ranged from US $0.74 and US $1.54, respectively, for Gram stain diagnosis, to US $8.32 and US $18.49 for the FemExam two-card method . CONCLUSIONS: In a setting where BV was frequently associated with vaginal discharge, the FemExam test compared favorably with conventional clinical diagnosis, and it has the advantage of being rapid, less subjective, and easily performed . Cutting its cost would provide wider accessibility in developing countries. J Microbiol Methods, 2003 Aug, 54(2), 153 - 64 Tracking of injected and resident (previously injected) bacterial cells in groundwater using ferrographic capture; Johnson WP et al.; A high-resolution bacterial tracking technique, ferrographic capture, was used to enumerate fluorescent-stained bacterial cells that were injected into groundwater during a field experiment . The goal of the experiment was to investigate whether detachment of previously injected stained resident cells attached to aquifer sediment was enhanced in the presence of the newly injected mobile cells . This injection was an improvement on past experiments in that the attached (resident) cells were stained, allowing their concentrations to be enumerated directly by ferrographic capture (upon detachment) . Contrary to expectations based on previous experiments, enhanced detachment of stained resident cells did not occur upon the arrival of injected cells . Consistent with previous experiments, however, was the observation of ephemeral increases in unstained cell concentrations coincident with the arrival of the stained injected cells . The ephemeral pulses of unstained cells were previously speculated to represent enhanced detachment of unstained indigenous cells in response to hydrodynamic collision with injected cells . The lack of enhanced detachment of stained resident cells in the present experiments indicates that increased concentrations of unstained cells may have occurred by mechanisms other than hydrodynamic collision . Visually observed variations in stain intensity indicated that increased unstained cell concentrations may have resulted from cell division at the low-concentration fringe of the injected plume. Eur J Obstet Gynecol Reprod Biol, 2003 Jun 10, 108(2), 146 - 51 Bacterial vaginosis: prevalence and predictive value for premature delivery and neonatal infection in women with preterm labour and intact membranes; Goffinet F et al.; OBJECTIVES: Assess the predictive values of bacterial vaginosis (BV) for preterm delivery (PD) and neonatal infection and compare them with standard markers of infection among women with preterm labour (PL) . STUDY DESIGN: Prospective blinded study in a tertiary referral centre in Paris . Women hospitalised for PL with intact membranes at a term between 24 and 34 weeks were included . Vaginal fluid, collected at inclusion was Gram-stained, scored, and interpreted according to Nugent's criteria . RESULTS: Out of 354 women tested, 254 had normal flora (72.3%), 76 intermediate (21.7%) and 24 BV (6.8%) . A history of spontaneous miscarriage after 14 weeks was the only risk factor significantly associated with BV . BV was not significantly associated with PD<35 weeks or neonatal infection . Very preterm delivery (before 33 weeks) was significantly associated with the flora grade (P=0.02): women with normal, intermediate and abnormal flora, respectively had 27 (10.6%), 14 (18.4%) and 6 (25.0%) births before 33 weeks . Of the markers tested, the highest risk of very preterm delivery was associated with BV (odds ratio 2.95, 95% CI (1.1-0.8.1)) and CRP>20mg/dl (4.23 95% CI (1.8-9.7)) . Predictive value of BV for preterm birth before 33 weeks were: sensitivity 12.8%, specificity 95.0%, positive predictive value 35.3%, and negative predictive value 84.3% . CONCLUSIONS: The frequency of BV and its association with PD are probably very variable and must be interpreted differently from one population to another . While we found an association between BV results and delivery before 33 weeks, the predictive value of BV was disappointing . Although these findings reinforce the importance of a useful marker of subclinical infection, the usefulness of testing for BV in women with PL has not been demonstrated. Arq Gastroenterol, 2002 Jul-Sep, 39(3), 158 - 62 Epub 2003 May 21. {Prevalence and prognosis of spontaneous bacterial peritonitis . Experience in patients from a general hospital in Porto Alegre, RS, Brazil (1991-2000)}; Coral G et al.; BACKGROUND: Spontaneous bacterial peritonitis is a frequent complication that occurs in patients with cirrhosis and ascites and has a recurrence rate of 70% in 1 year . In addition, this infection determines a poor short and long-term prognosis and a shorter survival rate . AIMS: Evaluate the prevalence of spontaneous bacterial peritonitis in cirrhotic patients with ascites and the effect of its occurrence on the survival . PATIENTS/METHODS: One thousand and thirty admissions of patients with cirrhosis and ascites were reviewed and 114 episodes of spontaneous bacterial peritonitis were documented in 94 patients . The ascitic analysis was accomplished in all patients . The diagnosis of this infection was established when the ascitic fluid polymorphonuclear count was equal or above 250 cells mm3 . RESULTS: The prevalence of this infection was 11.1% and the mortality rate 21.9% . Spontaneous bacterial peritonitis was community acquired in 61.4% and hospital acquired in 37.7% . The mortality rate was 18.6% and 27.9%, respectively . The infection resolved in 91.1% of the episodes by the analysis of ascitic fluid at 48 hours on antibiotics . The use of prophylactic antibiotics was documented in 22.3% of the episodes, but there are not significant differences on the mortality or type of bacteria isolated when comparing the patients with or without this treatment . CONCLUSIONS: Spontaneous bacterial peritonitis is a common complication in patients with cirrhosis and ascites and determines a worse prognosis, mainly when related with absence of initial response to antibiotics. J Trauma, 2003 May, 54(5), 908 - 14 Allogeneic blood transfusion increases the risk of postoperative bacterial infection: a meta-analysis; Hill GE et al.; BACKGROUND: Immunosuppression is a consequence of allogeneic (homologous) blood transfusion (ABT) in humans and is associated with an increased risk in cancer recurrence rates after potentially curative surgery as well as an increase in the frequency of postoperative bacterial infections . Although a meta-analysis has been reported demonstrating the relationship between ABT and colon cancer recurrence, no meta-analysis has been reported demonstrating the relationship of ABT to postoperative bacterial infection . METHODS: Twenty peer-reviewed articles published from 1986 to 2000 were included in a meta-analysis . Criteria for inclusion included a clearly defined control group (nontransfused) compared with a treated (transfused) group and statistical analysis of accumulated data that included stepwise multivariate logistic regression analysis . In addition, a subgroup of publications that included only the traumatically injured patient was included in a separate meta-analysis . A fixed effects analysis was conducted with odds ratios obtained by using the conditional maximum likelihood method and 95% confidence intervals on the obtained odds ratios were determined using the mid-p technique . RESULTS: The total number of subjects included in this meta-analysis was 13,152 (5,215 in the transfused group and 7,937 in the nontransfused group) . The common odds ratio for all articles included in this meta-analysis evaluating the association of ABT to the incidence of postoperative bacterial infection was 3.45 (range, 1.43-15.15), with 17 of the 20 studies demonstrating a value of p < or = 0.05 . These results provide overwhelming evidence that ABT is associated with a significantly increased risk of postoperative bacterial infection in the surgical patient . The common odds ratio of the subgroup of trauma patients was 5.263 (range, 5.03-5.43), with all studies showing a value of p < 0.05 (0.005-0.0001) . These results demonstrate that ABT is associated with a greater risk of postoperative bacterial infection in the trauma patient when compared with those patients receiving ABT during or after elective surgery . CONCLUSION: These results demonstrate that ABT is an associated and apparently significant and frequently overlooked risk factor for the development of postoperative bacterial infection in the surgical patient . Allogeneic blood transfusion is a greater risk factor in the traumatically injured patient when compared with the elective surgical patient for the development of postoperative bacterial infection. Acta Crystallogr D Biol Crystallogr, 2003 Jun, 59(Pt 6), 1061 - 3 Epub 2003 May 23. Crystallization of Hfq protein: a bacterial gene-expression regulator; Vassilieva IM et al.; Hfq protein from Escherichia coli (EcoHfq) has been overproduced in E . coli, purified to homogeneity and crystallized using the hanging-drop vapour-diffusion technique . Crystallization conditions for EcoHfq were found which yielded X-ray quality crystals . Crystals of EcoHfq and of Cd-, Hg- and Se-containing derivatives grew in two months, with unit-cell parameters a = b = 127.41, c = 170.36 A . The crystals belong to space group I4 and diffract to 2.1 A resolution . Two hexamers are predicted per asymmetric unit. Plant Mol Biol, 2003 Apr, 51(6), 803 - 15 Differential expression of genes encoding calmodulin-binding proteins in response to bacterial pathogens and inducers of defense responses; Ali GS et al.; Calmodulin (CaM) plays an important role in sensing and transducing changes in cellular Ca2+ concentration in response to several biotic and abiotic stresses . Although CaM is implicated in plant-pathogen interactions, its molecular targets and their role in defense signaling pathway(s) are poorly understood . To elucidate the signaling pathways that link CaM to defense responses, we screened a cDNA library constructed from bean leaves undergoing a hypersensitive response (HR) with radiolabeled CaM isoforms . A total of 26 putative CBPs were identified . Sequencing of the cDNAs revealed that they represent 8 different genes . They are homologues of previously identified CaM-binding proteins (CBPs) in other systems . However, some CBPs are novel members of known CBP families . The proteins encoded by these clones bound CaM in a Ca2+-dependent manner . To determine if these CBPs are involved in plant defense responses, we analyzed their expression in bean leaves inoculated with compatible, incompatible and nonpathogenic bacterial strains . Expression of three CBPs including an isoform of cyclic nucleotide-gated channels (PvCNGC-A) and two hypothetical proteins (PvCBP60-C and PvCBP60-D) was induced whereas the expression of two other isoforms of CNGCs (PvCNGC-B and PvCNGC-C) was repressed in response to incompatible pathogens . The expression of the rest, a small auxin up RNA (PvSAUR1) and two hypothetical proteins (PvCBP60-A and PvCBP60-B), was not changed . The expression of most of the pathogen-regulated genes was also affected by salicylic acid, jasmonic acid, hydrogen peroxide and a fungal elicitor, which are known to induce defense responses . Our results strongly suggest that at least five bean CBPs are involved in plant defense responses. J Bacteriol, 2003 Jun, 185(12), 3636 - 43 Electron microscopic analysis of membrane assemblies formed by the bacterial chemotaxis receptor Tsr; Weis RM et al.; The serine receptor (Tsr) from Escherichia coli is representative of a large family of transmembrane receptor proteins that mediate bacterial chemotaxis by influencing cell motility through signal transduction pathways . Tsr and other chemotaxis receptors form patches in the inner membrane that are often localized at the poles of the bacteria . In an effort to understand the structural constraints that dictate the packing of receptors in the plane of the membrane, we have used electron microscopy to examine ordered assemblies of Tsr in membrane extracts isolated from cells engineered to overproduce the receptor . Three types of assemblies were observed: ring-like "micelles" with a radial arrangement of receptor subunits, two-dimensional crystalline arrays with approximate hexagonal symmetry, and "zippers," which are receptor bilayers that result from the antiparallel interdigitation of cytoplasmic domains . The registration among Tsr molecules in the micelle and zipper assemblies was sufficient for identification of the receptor domains and for determination of their contributions to the total receptor length . The overall result of this analysis is compatible with an atomic model of the receptor dimer that was constructed primarily from the X-ray crystal structures of the periplasmic and cytoplasmic domains . Significantly, the micelle and zipper structures were also observed in fixed, cryosectioned cells expressing the Tsr receptor at high abundance, suggesting that the modes of Tsr assembly found in vitro are relevant to the situation in the cell. Rev Neurol (Paris), 2003 Apr, 159(4), 421 - 4 {Early diagnosis of bacterial brain abscesses: interest of diffusion-weighted MRI}; Detante O et al.; Three cases of bacterial brain abscesses, in immunocompetent patients, are reported . In all these cases, the diffusion-weighted magnetic resonance (MRI) with apparent diffusion coefficient (ADC) map has permitted an early diagnosis and a rapid treatment . This emergency MRI showed in the three cases a low signal on TI-weighted images, a high signal on T2-weighted and echo-planar images, and a decrease of ADC (0.36- 0.49 x 10(-3) mm2/s) . So, this new MRI technique provides an available and rapid element in the brain abscess diagnosis which often remains a complex clinical and radiological diagnosis. Toxicol Sci, 2003 Aug, 74(2), 457 - 69 Epub 2003 May 28. The coagulation system contributes to synergistic liver injury from exposure to monocrotaline and bacterial lipopolysaccharide; Yee SB et al.; Coexposure to a noninjurious dose of bacterial lipopolysaccharide (LPS; 7.4 x 106 EU/kg) and a nontoxic dose of the food-borne toxin monocrotaline (MCT; 100 mg/kg) leads to synergistic hepatotoxicity in Sprague-Dawley rats . Inflammatory factors, such as Kupffer cells (KCs), tumor necrosis factor-alpha (TNF)-alpha, and neutrophils (polymorphonuclear leukocytes; PMNs), are critical to the pathogenesis . Inasmuch as activation of the coagulation system and sinusoidal endothelial cell (SEC) injury precede hepatic parenchymal cell (HPC) injury, and since fibrin deposition occurs within liver lesions, the coagulation system might be a critical component of injury . In this study, this hypothesis is tested, and the interdependence of the coagulation system and inflammatory factors is explored . Administration of the anticoagulants heparin or warfarin to MCT/LPS-cotreated animals attenuated HPC and SEC injury . Morphometric analysis revealed that anticoagulant treatment significantly reduced the area of centrilobular and midzonal lesions . Heparin treatment also reduced fibrin deposition in these regions . Furthermore, anticoagulant treatment decreased hepatic PMN accumulation but did not affect plasma TNF-alpha concentration . Neither KC inactivation nor TNF-alpha depletion prevented activation of the coagulation system . PMN depletion, however, prevented coagulation system activation, suggesting that PMNs are needed for this response . These results provide evidence that the coagulation system and its interplay with PMNs are important in the pathogenesis of MCT/LPS-induced liver injury. Nucleic Acids Res . 2003 Jun 1;31(11):e65. An automated microplate-based method for monitoring DNA strand breaks in plasmids and bacterial artificial chromosomes; Rock C et al.; A method is described for high-throughput monitoring of DNA backbone integrity in plasmids and artificial chromosomes in solution . The method is based on the denaturation properties of double-stranded DNA in alkaline conditions and uses PicoGreen fluorochrome to monitor denaturation . In the present method, fluorescence enhancement of PicoGreen at pH 12.4 is normalised by its value at pH 8 to give a ratio that is proportional to the average backbone integrity of the DNA molecules in the sample . A good regression fit (r2 > 0.98) was obtained when results derived from the present method and those derived from agarose gel electrophoresis were compared . Spiking experiments indicated that the method is sensitive enough to detect a proportion of 6% (v/v) molecules with an average of less than two breaks per molecule . Under manual operation, validation parameters such as inter-assay and intra-assay variation gave values of <5% coefficient of variation . Automation of the method showed equivalence to the manual procedure with high reproducibility and low variability within wells . The method described requires as little as 0.5 ng of DNA per well and a 96-well microplate can be analysed in 12 min providing an attractive option for analysis of high molecular weight vectors . A preparation of a 116 kb bacterial artificial chromosome was subjected to chemical and shear degradation and DNA integrity was tested using the method . Good correlation was obtained between time of chemical degradation and shear rate with fluorescence response . Results obtained from pulsed- field electrophoresis of sheared samples were in agreement with those obtained using the microplate-based method. J Exp Med, 2003 Jun 2, 197(11), 1417 - 25 Epub 2003 May 27. Effect of anatomical distribution of mast cells on their defense function against bacterial infections: demonstration using partially mast cell-deficient tg/tg mice; Jippo T et al.; Mast cells were depleted in the peritoneal cavity of WBB6F1-tg/tg mice that did not express a transcription factor, MITF . When acute bacterial peritonitis was induced in WBB6F1-+/+, WBB6F1-W/Wv, and WBB6F1-tg/tg mice, the proportion of surviving WBB6F1-+/+ mice was significantly higher than that of surviving WBB6F1-W/Wv or WBB6F1-tg/tg mice . The poor survival of WBB6F1-W/Wv and WBB6F1-tg/tg mice was attributed to the deficient influx of neutrophils into the peritoneal cavity . The injection of cultured mast cells (CMCs) derived from WBB6F1-+/+ mice normalized the neutrophil influx and reduced survival rate in WBB6F1-W/Wv mice, but not in WBB6F1-tg/tg mice . This was not attributable to a defect of neutrophils because injection of TNF-alpha increased the neutrophil influx and survival rate in both WBB6F1-W/Wv and WBB6F1-tg/tg mice . Although WBB6F1-+/+ CMCs injection normalized the number of mast cells in both the peritoneal cavity and mesentery of WBB6F1-W/Wv mice, it normalized the number of mast cells only in the peritoneal cavity of WBB6F1-tg/tg mice . Mast cells within the mesentery or mast cells in the vicinity of blood vessels appeared to play an important role against the acute bacterial peritonitis . WBB6F1-tg/tg mice may be useful for studying the effect of anatomical distribution of mast cells on their antiseptic function. J Biotechnol, 2003 Jun 12, 103(1), 67 - 76 Effects of bacterial treatments on wood extractives; Kallioinen A et al.; Bacterial strains were isolated from spruce wood chips and their ability to reduce the content of wood extractives was studied . Strains were screened by cultivation on liquid media containing wood extractives as the major nutrient . Some bacterial species could decrease remarkably the amount of extractives in the liquid media and reduced the amount of triglycerides, steryl esters and total extractives by 100, 20 and 39%, respectively . Spruce wood chips were treated in controlled conditions with selected bacteria to test their effects on the chips . All the bacteria grew well on wood chips . The effect of bacterial metabolism on wood extractives was significant . Bacterial treatments reduced the amount of lipophilic extractives by 16-38% in 1 week of treatment and up to 67% in 2 weeks . The most efficient strain removed 90, 66 and 50% of triglycerides, steryl esters and resin acids, respectively, in 2 weeks . These results indicate that bacteria may be promising agents for the removal of extractives for improved pulping and papermaking processes. J Insect Physiol, 1998 Feb, 44(2), 157 - 164 The influence of bacterial species and intensity of infections on nodule formation in insects; Stanley DW et al.; Nodulation is the predominant cellular immune reaction to bacterial infection in insects . Nodulation is a complex process involving an unknown number of discrete cellular actions . Currently, there is only limited information on the signal transduction mechanisms that result in nodulation . In older larvae of the tobacco hornworm, Manduca sexta, and of the tenebrionid beetle, Zophobas atratus, eicosanoids are involved in one or more steps in the overall process, and treating these insects with inhibitors of eicosanoid biosynthesis prior to bacterial infection severely impairs their ability to form nodules . In this paper we address more detailed questions on eicosanoid-mediated nodulation . The nodulation reaction to bacterial infection occurs in all larval stages we examined, specifically, second, third, and fourth instars of M . sexta . In both species, the number of nodules formed in response to bacterial infection is related in an exponential way to the number of bacterial cells in the infection . Nodulation is also not related to larval size . We also found that nodulation intensity varies according to the species of infecting bacteria. DNA Repair (Amst), 2003 Jun 11, 2(6), 707 - 18 Comparative analysis of 8-oxoG:C, 8-oxoG:A, A:C and C:C DNA repair in extracts from wild type or 8-oxoG DNA glycosylase deficient mammalian and bacterial cells; Dantzer F et al.; We have investigated repair of DNA containing 8-oxoguanine and certain mismatches in cell-free extracts from mouse embryonic fibroblasts (MEFs) using a plasmid substrate with a single lesion at a defined position . Repair synthesis was monitored in a small restriction fragment with different size single strands in order to follow the fate of repair reactions in both strands at the same time . An important part of the study was to assess the role of OGG1 in various repair reactions and the experiments were carried out with extracts from mouse embryonic fibroblasts diploid for a mogg1 deletion (Ogg1(-/-)) as well as wild type . In wild type, DNA containing 8-oxoG:C was repaired in the expected fashion predominantly through short-patch repair . Overall repair was reduced to 20% in the Ogg1(-/-) extracts and to 40% if only long-patch repair was considered . The 8-oxoG:A pair was processed similarly in wild type and Ogg1(-/-) extracts and repair synthesis at A as well as at 8-oxoG could be demonstrated, however, to the same extent in Ogg1(-/-) and wild type for both strands . Extracts from Ogg1(-/-) behaved normally in the correction of A:C and C:C mismatches, with a strong bias for correction of A for A:C and no significant strand discrimination for C:C . Similar experiments with extracts from Escherichia coli showed a 50% reduction in the repair of 8-oxoG:C in fpg extracts and an increase to 50% above wild type in mutY . These results show that the mouse OGG1 is the major enzyme for 8-oxoG repair in the MEF cells and does not participate in mismatch repair of A:C or C:C . Furthermore, 8-oxoG opposite A appears to be repaired by a two-step repair pathway with sequential removal of A and 8-oxoG mediated by enzymes different from OGG1. Water Res, 2003 Jul, 37(12), 2929 - 36 Bacterial immobilization and oxidation of arsenic in acid mine drainage (Carnoulès creek, France); Casiot C et al.; The acid waters (pH=2.73-3.37) originating from the Carnoules mine tailings contain high dissolved concentrations of arsenic (1-3.5 mmol l(-1)) and iron (20-40 mmol l(-1)) . At the outlet, arsenite predominates . During the first 30 m of downflow, 20-60% is removed by coprecipitation with Fe(III) . This process results from bacterially mediated As- and Fe-oxidation . The precipitation rates in the creek depend on the oxygen concentration in spring water and are lower during the dry summer period when the anoxic character of the spring water inhibits the activity of oxidizing bacteria . Ex situ experiments show that the presence of bacteria-rich precipitates increases the As- and Fe-removal rates . Three strains of bacteria promoting the oxidation of As have been isolated, and two of them have the characteristics of Thiomonas ynys1 . The third strain, which is not identified yet, also catalyzes the oxidation of Fe. Clin Infect Dis, 2003 Jun 1, 36(11), 1492 - 5 Epub 2003 May 16. Clearance of 14-3-3 protein from cerebrospinal fluid heralds the resolution of bacterial meningitis; Bonora S et al.; The 14-3-3 protein, a cerebrospinal fluid (CSF) marker of neuronal damage that was recently adopted for the diagnosis of Creutzfeldt-Jakob disease, is also found in the CSF of patients with a variety of neurological disorders . We prospectively studied 12 consecutive patients with purulent bacterial meningitis and found that 14-3-3 protein was detected in all patients at admission to the hospital . All patients who recovered cleared 14-3-3 protein from the CSF before discharge from the hospital (this was the first CSF marker to clear), whereas those who died never cleared the protein. Clin Nutr, 2003 Jun, 22(3), 277 - 81 The effect of immunonutrition on bacterial translocation, and intestinal villus atrophy in experimental obstructive jaundice; Zulfikaroglu B et al.; BACKGROUND & AIMS: Spontaneous bacterial infection and septicemia due to increased bacterial translocation (BT) in patients with obstructive jaundice result in significant morbidity and mortality . The present study evaluates the effects of enteral nutrition with immune enhancing feeds on BT and intestinal villus histopathology promoted by obstructive jaundice . METHODS: Fifty male Wistar-albino rats weighing 250-300g were assigned into five equal groups of 10 . Animals in Groups I, II, and III were fed with standard chow, those in Group IV were given glutamine 1g/kg/day and the remaining 10 animals in Group V were fed with an arginine, omega-3 fatty acids, and RNA-supplemented enteral diet for (1g/kg/day amino acid and 230 kcal/kg) 7 days preoperatively . Group I underwent sham operation and the remaining animals in all other groups underwent common bile duct ligation . After operation, Group I had standard chow, Groups II and IV had glutamine, Groups III and V had an arginine omega-3 fatty acids, and RNA-supplemented enteral diet for 7 days . All animals were sacrificed on the 8th postoperative day and evaluated both biochemically and histopathologically . Samples from blood, liver, mesenteric lymph nodes and spleen were cultured under aerobic conditions . RESULTS: Significantly less BT was observed in groups fed with an arginine, omega-3 fatty acids, and RNA-supplemented enteral diet or glutamine in pre-and postoperative periods as compared to others (P<0.001) . Histologic evaluation also showed significant reduction in villus atrophy in these groups . CONCLUSIONS: Enteral immunonutrition using glutamine or arginine, omega-3 fatty acids, and RNA-supplemented enteral diet during both pre-and postoperative periods seems to reduce BT and decrease atrophy of intestinal mucosal villi in rats with obstructive jaundice. In Silico Biol, 2003, 3(1-2), 127 - 43 Epub 2003 Mar 16. Analysis of bacterial RM-systems through genome-scale analysis and related taxonomy issues; Vandenbogaert M et al.; Recognition sites for type II restriction and modification enzymes in genomes of several bacteria are recognized as semi-palindromic motifs and