J Biol Chem, 2001 Dec 7, 276(49), 45513 - 5 Epub 2001 Oct 16. Revealing the involvement of extended hydrogen bond networks in the cooperative function between distant sites in bacterial reaction centers; Tandori J et al.; In reaction center proteins of photosynthetic bacteria, the amplitude of proton uptake induced by the one-electron reduction of either of the two quinone electron acceptors (Q(A) and Q(B)) is an intrinsic observable of the electrostatic interactions associated with the redox function of the complex . We report here that, in Rhodobacter capsulatus, complete restoration of proton uptake (upon formation of Q(A)(-) and Q(B)(-)) to the level found in the wild type is observed in a mutant reaction center in which a tyrosine substitution in the Q(A) environment (Ala(M274) --> Tyr) is coupled with mutations of acidic residues near Q(B) (Glu(L212) --> Ala/Asp(L213) --> Ala) that initially cancel the proton uptake above pH 8 . This result demonstrates that proton uptake occurs by strong cooperation between structural motifs, such as hydrogen-bonded networks, that span the 18 A distance between the two quinone acceptors. Curr Opin Biotechnol, 2001 Oct, 12(5), 467 - 72 Transfer of eukaryotic expression plasmids to mammalian host cells by bacterial carriers; Weiss S et al.; The concept of transkingom transfer of DNA from bacteria to other organisms has recently been extended to include eukaryotic host cells . Intracellular bacteria have been shown to transfer eukaryotic expression plasmids to mammalian host cells in vitro and in vivo . This can be used to induce immune responses towards protein antigens encoded by the plasmid, to complement genetic defects or even to direct the production of proteins in appropriate organs . The ease of generating such vehicles makes this a highly attractive area for further research. Ann Thorac Surg, 2001 Oct, 72(4), 1327 - 30 Autologous transfusion of shed mediastinal blood after coronary artery bypass grafting and bacterial contamination; Andreasen AS et al.; BACKGROUND: Autologous transfusion of shed mediastinal blood is often used after coronary artery bypass grafting (CABG) . Shed blood has in a few studies been cultured during the first postoperative hours . However, autologous transfusion might in some cases be continued for several hours and no study has yet examined the bacterial contamination of shed blood later than 6 hours postoperatively . METHODS: Seventy-five patients undergoing electively performed CABG were included . Cultures of shed blood were taken at initiation of the autologous transfusion and the following morning . Infection variables were measured preoperatively and postoperatively . Infectious complications during the first postoperative week were registered . RESULTS: The frequency of patients with bacterial growth in the first culture was 0.22 (95% confidence interval: 0.12 to 0.31) compared with 0.04 (95% confidence interval: -0.044 to 0.087) in the second culture (p < 0.002) . We found no significant difference in infection variables between patients with or without bacterial growth in the cultures . No patients suffered from early postoperative infectious complications . CONCLUSIONS: There is no further contamination of the shed blood during the period between initiating the autologous transfusion and the following morning. Urology . 2001 Oct;58(4):607. Massive hematuria due to right renal artery mycotic pseudoaneurysm in a patient with subacute bacterial endocarditis; Ohebshalom MM et al.; A 40-year-old woman with recently diagnosed bacterial endocarditis was admitted to the hospital with gross hematuria and anemia . Computed tomography revealed a large right upper pole renal artery pseudoaneurysm, a wedge-shaped hypoperfused region of the left kidney, and a splenic abscess . Radiographic embolization of the right renal artery was performed to stabilize the bleeding . The splenic abscess was drained . Subsequent right nephrectomy and splenectomy were performed for persistent leukocytosis . This unusual presentation of a septic embolus and its management are discussed. Biochim Biophys Acta, 2001 Nov 1, 1515(1), 44 - 54 Encapsulation of DNA in negatively charged liposomes and inhibition of bacterial gene expression with fluid liposome-encapsulated antisense oligonucleotides; Fillion P et al.; Antisense therapy for the treatment of bacterial infections is a very attractive alternative to overcome drug resistance problems . However, the penetration of antisense oligonucleotides into bacterial cells is a major huddle that has delayed research and application in this field . In the first part of this study, we defined efficient conditions to encapsulate plasmid DNA and antisense oligonucleotides in a fluid negatively charged liposome . Subsequently, we evaluated the potential of liposome-encapsulated antisense oligonucleotides to penetrate the bacterial outer membrane and to inhibit gene expression in bacteria . It was found that 48.9+/-12% and 43.5+/-4% of the purified plasmid DNA and antisense oligonucleotides were respectively encapsulated in the liposomes . Using fluorescence-activated cell sorting analysis, it was shown, after subtraction of the fluorescence values due to the aggregation phenomenon measured at 4 degrees C, that about 57% of bacterial cells had integrated the encapsulated antisense oligonucleotides whereas values for free antisenses were negligible . The uptake of the encapsulated anti-lacZ antisense oligonucleotides resulted in a 42% reduction of beta-galactosidase compared to 9% and 6% for the encapsulated mismatch antisense oligonucleotides and the free antisense oligonucleotides respectively . This work shows that it is possible to encapsulate relatively large quantities of negatively charged molecules in negative fluid liposomes and suggests that fluid liposomes could be used to deliver nucleic acids in bacteria to inhibit essential bacterial genes. Plant Cell, 2001 Oct, 13(10), 2257 - 68 Plastid division is driven by a complex mechanism that involves differential transition of the bacterial and eukaryotic division rings; Miyagishima Sy et al.; During plastid division, two structures have been detected at the division site in separate analyses . The plastid-dividing ring can be detected by transmission electron microscopy as two (or three) electron-dense rings: an outer ring on the cytosolic face of the outer envelope, occasionally a middle ring in the intermembrane space, and an inner ring on the stromal face of the inner envelope . The FtsZ ring, which plays a central role in bacterial division, also is involved in plastid division and is believed to have descended to plastids from cyanobacterial endosymbiosis . The relationship between the two structures is not known, although there is discussion regarding whether they are identical . Biochemical and immunocytochemical investigations, using synchronized chloroplasts of the red alga Cyanidioschyzon merolae, showed that the plastid FtsZ ring is distinct and separable from the plastid-dividing ring . The FtsZ ring localizes in stroma and faces the inner plastid-dividing ring at the far side from the inner envelope . The FtsZ ring and the inner and outer plastid-dividing rings form in that order before plastid division . The FtsZ ring disappears at the late stage of constriction before dissociation of the plastid-dividing ring, when the constriction is still in progress . Our results suggest that the FtsZ ring;-based system, which originated from a plastid ancestor, cyanobacteria, and the plastid-dividing ring;-based system, which probably originated from host eukaryotic cells, form a complex and are involved in plastid division by distinct modes. J Physiol Paris, 2001 Jan-Dec, 95(1-6), 443 - 51 Effects of dexamethasone and FK506 on Helicobacter pylori-induced gastritis and bacterial viability in Mongolian gerbils; Naka Y et al.; FK506 and dexamethasone were used to investigate whether or not immunosuppression affects H . pylori colonization and gastric mucosal damage induced by Helicobacter pylori in Mongolian gerbils . Two weeks after H . pylori infection, FK506 and dexamethasone or vehicle alone were subcutaneously administered once daily for the following 2 weeks . FK506 or vehicle alone was administered subcutaneously once daily for 5 weeks (1 week before and 4 weeks after infection) . In H . pylori-infected animals for 4 weeks, hemorrhagic erosions and inflammatory responses (neutrophil infiltration and lymphoid follicle formation) were induced in gastric mucosa at an incidence of 100% . Both FK506 and dexamethasone administered for 2 weeks markedly reduced such mucosal changes . In these animals, H . pylori viability in the stomach was significantly elevated . FK506 administered for 5 weeks also significantly inhibited the hemorrhagic erosions, edema and neutrophil infiltration in the stomach . H . pylori viability was slightly elevated as compared with the control . It was concluded that the host immune responses might play dual roles both by deteriorating gastritis induced by H . pylori and by protecting against H . pylori infection in its early stage. Biol Chem, 2001 Aug, 382(8), 1271 - 6 Photo-CIDNP 13C magic angle spinning NMR on bacterial reaction centres: exploring the electronic structure of the special pair and its surroundings; Matysik J et al.; Photochemically induced dynamic nuclear polarisation (photo-CIDNP) in intact bacterial reaction centres has been observed by 13C-solid state NMR under continuous illumination with white light . Strong intensity enhancement of 13C NMR signals of the aromatic rings allows probing the electronic ground state of the two BChl cofactors of the special pair at the molecular scale with atomic selectivity . Differences between the two BChl cofactors are discussed . Several aliphatic 13C atoms of cofactors, as well as 13C atoms of the imidazole ring of histidine residue(s), show nuclear-spin polarisation to the same extent as the aromatic nuclei of the cofactors . Mechanisms and applications of polarisation transfer are discussed. J Immunol, 2001 Oct 15, 167(8), 4644 - 50 Meningeal and perivascular macrophages of the central nervous system play a protective role during bacterial meningitis; Polfliet MM et al.; Meningeal (MM) and perivascular macrophages (PVM) constitute major populations of resident macrophages in the CNS that can be distinguished from microglial cells . So far, there is no direct evidence that demonstrates a possible role of MM and PVM in the CNS during normal or pathologic conditions . To elucidate the role of the MM and PVM during CNS inflammation, we have developed a strategy using a single intraventricular injection of mannosylated clodronate liposomes, which results in a complete and selective depletion of the PVM and MM from the CNS . Depletion of the MM and PVM during experimental pneumococcal meningitis resulted in increased illness, which correlated with higher bacteria counts in the cerebrospinal fluid and blood . This was associated with a decreased influx of leukocytes into the cerebrospinal fluid, which occurred despite an elevated production of relevant chemokines (e.g., macrophage-inflammatory protein-2) and a higher expression of vascular adhesion molecules (e.g., VCAM-1) . In contrast, the higher bacterial counts correlated with elevated production of local and systemic inflammatory mediators (e.g., IL-6) indicating enhanced local leukocyte and systemic immune activation, and this may explain the worsening of the clinical signs . These findings show that the PVM and MM play a protective role during bacterial meningitis and suggest that a primary action of these macrophages is to facilitate the influx of leukocytes at the blood-brain barrier . More in general, we demonstrate for the first time that the PVM and MM play a crucial role during inflammation in the CNS. Chest, 2001 Oct, 120(4), 1377 - 89 Bacterial/viral filtration: let the breather beware! Demers RR. Most clinicians believe that any device that is marketed as a "bacterial/viral filter" must necessarily be capable of capturing any individual bacteria or viruses that might be suspended within inhaled or exhaled gases . We were surprised to discover that this is, by no means, a justifiable assumption . This article describes testing methods that manufacturers employ to generate the often-misleading efficiency specifications that are claimed for some of these devices . We discuss articles that have documented the presence of airborne pathogens in the effluent of a ventilator circuit, and characterize the attributes that a competent filter must exhibit if it is to succeed in protecting patients and caregivers from incidental exposure to bacteria, viruses, aerosolized drugs, and endotoxins . This article continues with a discussion of the numbers of particles that are commonly produced with commercially available pneumatic nebulizers, the comparative performance characteristics of filters and heat/moisture exchanging filters (HMEFs), and the success or failure of various brands of HMEFs to comply with the guidelines recently developed by the Centers for Disease Control and Prevention for the management of patients who are harboring active tuberculosis . The presentation concludes with a description of the standards that apply to any filter that classifies as a high-efficiency particulate aerosol (HEPA) device, and demonstrates that the performance of filters/HMEFs in common clinical use can range from approximately 1/50th to > 30-fold the efficiency of a HEPA-grade device . Those who frequent the bedside of patients receiving ventilation might unwittingly be placing themselves at considerable risk of exposure to infectious microaerosols, but methods are available to dramatically decrease those risks. Pancreas, 2001 Oct, 23(3), 296 - 301 Inhibition of inducible nitric oxide synthase reduces bacterial translocation in a rat model of acute pancreatitis; Simsek I et al.; INTRODUCTION: Translocation of bacteria from the gut into pancreatic necrosis is an important factor in the development of septic complications and mortality in acute pancreatitis . S-methylisothiourea (SMT) is an inducible nitric oxide synthase inhibitor that has been shown to decrease bacteria} translocation in sepsis and thermal injury . AIM: To investigate whether SMT could affect bacterial translocation in acute necrotizing pancreatitis . METHODOLOGY: Forty-five Sprague-Dawley rats were studied . Acute pancreatitis was induced in Group I and Group II by injection of taurocholate and trypsin into the common biliopancreatic duct . Group III underwent laparotomy with the manipulation (but not cannulation) of the pancreas and received saline injection . Group I rats received normal saline as a placebo, and Group II rats received SMT after surgery for 2 days . At 48 hours, blood was drawn for serum amylase determinations . Bacterial translocation to mesenteric lymph nodes and distant sites (pancreas, liver, and peritoneum) were examined . A point scoring system of histologic features was used to evaluate the severity of pancreatitis . RESULTS: Plasma amylase levels and pancreatic histologic score were significantly reduced in Group II rats given SMT compared with those in Group I rats given saline (p < 0.01, p < 0.05, respectively) . All Group I rats had bacterial translocation to mesenteric lymph nodes compared with 7 of 12 rats in Group II (p < 0.05) . There was no difference in bacterial translocation to distant organs between the two groups, although rates tended to be lower in Group II compared with Group I (p > 0.05) . Bacterial counts in the pancreas were significantly reduced in Group II rats compared with those in Group I rats (p < 0.05) . CONCLUSION: Treatment with SMT appears to have ameliorated the course of acute pancreatitis; however, mortality was not affected. Bioinformatics, 2001 Sep, 17(9), 791 - 802 Comprehensive comparison between locations of orthologous genes on archaeal and bacterial genomes; Horimoto K et al.; MOTIVATION: Following an extensive search for orthologous genes between the complete genomes from archaea and bacteria, the spatial association of the orthologs has been investigated in terms of synteny, the conservation of the order of neighboring genes . However, the relationships between the relative locations of remote orthologs over entire genomes have not been shown . RESULTS: Comprehensive comparisons between the locations of orthologs on nineteen archaeal and bacterial genomes are presented by the location to location correspondence based on the gene-location distance . When the two genomes are rotated such that a pair of orthologs with the shortest distance is set in the same angle, a statistically significant number of orthologs maintain their relative locations between the genomes . Even by the short distances at the 5% significance level, the rotations are restricted within a narrow range, suggesting an intrinsic angle for realizing similar locations between the orthologs in each genome pair . Furthermore, the rotations in the restricted range agree with the replication origin and terminus sites for the analyzed genomes where such sites are known . The relationship between location-maintained orthologs and gene function is also discussed. Trends Biochem Sci, 2001 Oct, 26(10), 579 - 82 The CHASE domain: a predicted ligand-binding module in plant cytokinin receptors and other eukaryotic and bacterial receptors; Anantharaman V et al.; A novel, 200-230 amino acid extracellular domain was identified in the plant cytokinin receptor Cre1, in the receptor-histidine kinase DhkA and the adenylyl cyclase Acg from the slime mold Dictyostelium discoideum, and in a variety of other receptor-like proteins from bacteria and eukaryotes . The domain is predicted to bind diverse low molecular weight ligands, such as the cytokinin-like adenine derivatives or peptides, and mediate signal transduction through the respective receptors. J Clin Gastroenterol, 2001 Oct, 33(4), 295 - 8 Long-term prognosis of cirrhosis after spontaneous bacterial peritonitis treated with ceftriaxone; Franca AV et al.; Spontaneous bacterial peritonitis (SBP) is a frequent infection in cirrhotic patients with ascites, with a poor prognosis . The aims of this study were to determine the long-term survival of cirrhotic patients with SBP treated with ceftriaxone and to identify predictive factors related to survival . We studied 47 first episodes of SBP treated with ceftriaxone with a mean follow-up of 272 days . Nineteen variables were recorded to evaluate their relation to survival . The most frequent organism that caused SBP was Escherichia coli (40%) . Spontaneous bacterial peritonitis resolution was achieved in 67% of patients . After resolution, SBP recurrence was observed in 44% of patients . The cumulative probability of survival was 68.1% at 1 month and 30.8% at 6 months . After uni-and multivariate analyses of all cases, SBP resolution ( p = 0.0001) and international normalized ratio (INR) ( p = 0.0057) were found to be related to survival . Another analysis performed after SBP resolution and SBP recurrence showed that ascitic fluid-positive culture ( p = 0.0344) and INR ( p = 0.0218) had statistical significance as variables predictive of long-term survival . We conclude that the survival of cirrhotic patients is very short after the first episode of SBP, a fact probably related to advanced liver disease, as liver dysfunction (INR) is the most important factor related to long-term patient survival. Curr Opin Microbiol, 2001 Oct, 4(5), 602 - 6 The relative contributions of recombination and point mutation to the diversification of bacterial clones; Spratt BG et al.; Low levels of recombination in bacterial species have often been inferred from the presence of linkage disequilibrium between the alleles at different loci in the population . However, significant linkage disequilibrium is inevitable in organisms that divide by binary fission, and recombinational replacements must be very frequent, compared to point mutation, to dissipate disequilibrium . Recent studies using data from multilocus sequence typing indicate that, in many species, recombinational replacements contribute more greatly to clonal diversification than do point mutations and, in some species, recombination has been sufficient to eliminate any phylogenetic signal from gene trees . Recent efforts to improve understanding of the extent and impact of homologous recombination in the diversification of bacterial clones are discussed. Trends Genet, 2001 Oct, 17(10), 589 - 96 Deletional bias and the evolution of bacterial genomes; Mira A et al.; Although bacteria increase their DNA content through horizontal transfer and gene duplication, their genomes remain small and, in particular, lack nonfunctional sequences . This pattern is most readily explained by a pervasive bias towards higher numbers of deletions than insertions . When selection is not strong enough to maintain them, genes are lost in large deletions or inactivated and subsequently eroded . Gene inactivation and loss are particularly apparent in obligate parasites and symbionts, in which dramatic reductions in genome size can result not from selection to lose DNA, but from decreased selection to maintain gene functionality . Here we discuss the evidence showing that deletional bias is a major force that shapes bacterial genomes. Mutat Res, 2001 May 10, 485(4), 331 - 8 Characterization of functional interactions among the Escherichia coli mismatch repair proteins using a bacterial two-hybrid assay; Mansour CA et al.; Vsr mediates very short patch repair in Escherichia coli, correcting T/G mismatches caused by deamination of 5-methylcytosine to thymine . MutS and MutL, part of the post-replication mismatch repair system, stimulate VSP repair . In this study, we use a bacterial two-hybrid assay to show that MutL interacts with Vsr . We also show that interaction between Vsr and MutL inhibits the ability of MutL to dimerize, to interact with MutS and MutH and to mediate a previously unknown interaction between MutS and MutH . This inhibition may explain why high levels of Vsr are mutagenic in vivo . In addition, we show that the Mut fusion proteins are repair proficient in the bacterial two-hybrid assay, making it possible to study their interactions in various genetic backgrounds, or in the presence of DNA damaging agents. Biochemistry, 2001 Oct 9, 40(40), 12132 - 9 Manipulating the direction of electron transfer in the bacterial reaction center by swapping Phe for Tyr near BChl(M) (L181) and Tyr for Phe near BChl(L) (M208); Kirmaier C et al.; We have investigated the primary charge separation processes in Rb . capsulatus reaction centers (RCs) bearing the mutations Phe(L181) --> Tyr, Tyr(M208) --> Phe, and Leu(M212) --> His . In the YFH mutant, decay of the excited primary electron donor P occurs with an 11 +/- 2 ps time constant and is trifurcated to give (1) internal conversion to the ground state ( approximately 10% yield), (2) charge separation to the L side of the RC ( approximately 60% yield), and (3) electron transfer to the M-side bacteriopheophytin BPh(M) ( approximately 30% yield) . These results relate previous work in which the ionizable residues Lys (at L178) and Asp (at M201) have been used to facilitate charge separation to the M side of the RC, and the widely studied L181 and M208 mutants . One conclusion that comes from this work is that the Tyr (M208) --> Phe and Gly(M201) --> Asp mutations near the L-side bacteriochlorophyll (BChl(L)) raise the free energy of P(+)BChl(L)(-) by comparable amounts . The results also suggest that the free energy of P(+)BChl(M)(-) is lowered more substantially by a Tyr at L181 than a Lys at L178 . The results on the YFH mutant further demonstrate that the free energy differences between the L- and M-side charge-separated states play a significant role in the directionality of charge separation in the wild-type RC, and place limits on the contributing role of differential electronic matrix elements on the two sides of the RC. An Esp Pediatr, 2001 Oct, 55(4), 321 - 8 {Procalcitonin in the early diagnosis of invasive bacterial infection in febrile infants}; Fernandez Lopez A et al.; BACKGROUND: Procalcitonin (PCT) it is a new marker of bacterial infection . Because of its shorter half-life and earlier ascent it offers advantages over C-reactive protein (CRP) . OBJECTIVE: To compare the diagnostic performance of PCT in the early detection of invasive bacterial infection in infants with that of CPR . MATERIAL AND METHODS: Between January of 1998 and February of 2000 we performed a prospective observational study in the emergency department of infants aged between 1 and 36 months who had been treated for fever and for whom PCT and CRP plasmatic values had been obtained . Plasmatic PCT and PCR values were evaluated and correlated with the final diagnosis . ROC curves for both markers were calculated . RESULTS: One hundred infants with a mean age of 8.8 months (SD 7.59) were included in four groups of 25 patients each (viral infection, localized bacterial infection, invasive bacterial infection and control group) . The mean PCT and CRP values in invasive bacterial infections {PCT: 14.45 ng/mL (SD 27.95) and CRP: 95.10 mg/L (SD 7 2.77)} were significantly higher than in non-invasive infections {PCT: 0.27 ng/mL (SD 0.19) and CRP: 25.67 mg/L (SD 33.04)} but the diagnostic performance of PCT was better . The area under the curve for PCT was 0.95 (SD 0.03), which was significantly higher (p < 0.001) than that obtained for CRP {0.81 (SD 0.05)} . The optimal cut-off for PCT was > 0.4 ng/mL (sensitivity: 95.5 %; specificity: 86.4 %) and that for CRP was > 42.9 mg/L (sensitivity: 75 %; specificity: 81.8 %) . In infants who had fever for less than 12 hours (n 30) the area under the curve for PCT was 0.90 (SD 0.06), which was higher (p < 0.001) than that for PCR {0.64 (SD 0.11)} . The optimal cut-off for PCT in this group was > 0.4 ng/mL (sensitivity: 90 %; specificity: 94 %) and that for CRP was > 26.6 mg/L (sensitivity: 60 %; specificity: 77.8 %) . CONCLUSIONS: The diagnostic performance of PCT was higher than that of CRP in the early detection of invasive infection in febrile infants, even when evolution was less than 12 hours. Clin Infect Dis, 2001 Nov 1, 33(9), e109 - 11 Epub 2001 Sep 26. Cerebrospinal fluid penetration of levofloxacin in patients with spontaneous acute bacterial meningitis; Scotton PG et al.; We have assessed levofloxacin penetration in cerebrospinal fluid (CSF) and the liquor-to-plasma ratio (C(L)/C(P)) at 2 hours after dosing in 5 patients with spontaneous acute bacterial meningitis . CSF levofloxacin concentration at 2 hours after dosing was 1.99+/-0.67 microg/mL, and the C(L)/C(P) at 2 hours after dosing was 0.34+/-0.09. J Microbiol Methods, 2001 Nov, 47(2), 219 - 31 Comparison of methods for monitoring bacterial transport in the subsurface; DeFlaun MF et al.; The purpose of this study was to compare in a laboratory experiment, a suite of methods developed to track viable bacteria during field transport experiments . The criteria for development and selection of these methods included: (1) the ability to track bacteria within the environment from which they were isolated; (2) the lack of any effect upon the viability or the transport characteristics of the strain; (3) low detection limits; (4) a quantification range that covered several orders of magnitude; and (5) an analytical cost and turnover time commensurate with the analysis of several thousands of samples in a few months . The approaches developed included: enumeration of bacteria labeled with a vital fluorescent stain (CFDA/SE) using microplate spectrofluorometry, flow cytometry, and ferrographic (immunomagnetic) capture; enumeration of highly (13)C-enriched bacteria using combustion-IRMS; and quantitative PCR . These methods were compared to direct microscopic enumeration and plate counts during a bacterial transport experiment performed in an intact sediment core and designed to simulate the field experiment . Four of the seven methods had equivalent recoveries for the breakthrough of a pulse of bacteria eluting from a 50-cm long sediment core, and all of the methods detected the arrival of cells in the effluent prior to the conservative tracer . Combustion IRMS and ferrographic enumeration had the lowest quantification limits (approximately 2 to 20 cells/ml), whereas microplate spectrofluorometry had the highest quantification limit (approximately 10(5) cells/ml) . These methods have the potential for numerous applications beyond tracking bacteria injected into the subsurface. Obstet Gynecol, 2001 Oct, 98(4), 656 - 63 Longitudinal analysis of bacterial vaginosis: findings from the HIV epidemiology research study; Jamieson DJ et al.; OBJECTIVE: To determine the natural history of bacterial vaginosis in women with or at risk for human immunodeficiency virus (HIV) . METHODS: A cohort of 854 HIV-infected women and 434 HIV-uninfected women from four US sites was followed prospectively with gynecologic exams every 6 months over a 5-year period . The prevalence, incidence, persistence, and severity of bacterial vaginosis, which was defined using a Gram-staining scoring system, were calculated using generalized estimating equation methods . RESULTS: In adjusted analyses, HIV-infected women had a higher prevalence of bacterial vaginosis than HIV-uninfected women (adjusted odds ratio {OR} 1.29; 95% confidence interval {CI} 1.08, 1.55) . Although HIV-infected women were not more likely to have incident infections, they were more likely to have persistence of their infections (adjusted OR 1.49; 95% CI 1.18, 1.89) . Similarly, immunocompromised women (CD4+ cell count less than 200 cells/microL) were more likely than HIV-infected women with higher CD4+ cell counts (more than 500 cells/microL) to have prevalent (adjusted OR 1.29; 95% CI 1.03, 1.60) and persistent (adjusted OR 1.38; 95% CI 1.01, 1.91) bacterial vaginosis infections, but not more likely to have incident infections . Immunocompromised women had more severe bacterial vaginosis by both clinical criteria (adjusted OR 1.40; 95% CI 1.08, 1.82) and by Gram-staining criteria (adjusted OR 1.50; 95% CI 1.12, 2.00) . CONCLUSIONS: Bacterial vaginosis is more prevalent and persistent among HIV-infected women, particularly among those who are immunocompromised . Immunocompromised women are more likely than HIV-infected women with higher CD4+ cell counts to have severe bacterial vaginosis. FEBS Lett, 2001 Sep 21, 505(3), 353 - 6 The central plug in the reconstituted undecameric c cylinder of a bacterial ATP synthase consists of phospholipids; Meier T et al.; The isolated rotor cylinder of the ATP synthase from Ilyobacter tartaricus was reconstituted into two-dimensional crystalline arrays . Atomic force microscopy imaging indicated a central cavity on one side of the rotor and a central plug protruding from the other side . Upon incubation with phospholipase C, the plug disappeared, but the appearance of the surrounding c subunit oligomer was not affected . This indicates that the plug consists of phospholipids . As the detergent-purified c cylinder is completely devoid of phospholipids, these are incorporated into the central hole from one side of the cylinder during the reconstitution procedure. Am J Public Health, 2001 Oct, 91(10), 1664 - 70 Factors linked to bacterial vaginosis in nonpregnant women; Holzman C et al.; OBJECTIVES: The purposes of this study were to test the hypothesis that vaginal douching is linked to bacterial vaginosis in both symptomatic and asymptomatic women and to identify other demographic, reproductive, and lifestyle factors associated with bacterial vaginosis . METHODS: In this cross-sectional study involving 3 clinic sites, 496 nonpregnant women completed a self-administered questionnaire . Their vaginal smears were assessed and cross-validated for bacterial vaginosis . RESULTS: The prevalence of bacterial vaginosis across clinics ranged from 15% to 30% . In analyses restricted to site 1, adjusted odds ratios (ORs) for bacterial vaginosis remained significant for African American women with 13 or fewer years of education (OR = 5.5, 95% confidence interval {CI} = 2.1, 14.5), hormone use within the past 6 months (OR = 0.5, 95% CI = 0.2, 0.8), and vaginal douching within the past 2 months (OR = 2.9, 95% CI = 1.5, 5.6) . CONCLUSIONS: Two lifestyle factors emerge as strongly associated with bacterial vaginosis: systemic contraceptives appear protective, whereas douching is linked to an increase in prevalence . The temporal relationship between douching and bacterial vaginosis needs further clarification. Matern Child Health J, 2001 Jun, 5(2), 127 - 34 Maternal stress is associated with bacterial vaginosis in human pregnancy; Culhane JF et al.; OBJECTIVES: Maternal infection, particularly bacterial vaginosis (BV) in pregnancy, is one of the leading causes of adverse perinatal outcomes . The determinants of individual differences in susceptibility, or vulnerability, to maternal infections are poorly understood . This study examines whether chronic maternal stress predisposes women to infection during pregnancy, and if so, whether the effects of chronic stress on infection are independent of other established risk factors . METHODS: We conducted a cross-sectional, clinical prevalence study of chronic maternal stress and BV status in a sample of 454 pregnant women at 14.3+/-0.3 weeks gestation (+/-SEM) . BV was diagnosed by Gram-stain of vaginal fluid samples; chronic maternal stress was assessed using the Cohen Perceived Stress Scale . Other established risk factors for BV, including maternal age, race/ethnicity, marital status, SES, and behaviors related to feminine hygiene, sexual practices, and substance use, were measured using a structured interview . RESULTS: Of the 454 women enrolled in this study, 224 (49%) were BV positive (Nugent score 7-10), 64 (14%) had intermediate vaginal flora (Nugent score 4-6), and 166 (37%) were BV negative (Nugent score 0-3) . BV+ women had significantly higher chronic stress levels than BV- women (24.6+/-0.5 vs . 22.2+/-0.6 units (+/-SEM), respectively; t = 3.19; p < .01) . Maternal sociodemographic variables (African-American race/ethnicity) and behavioral characteristics (vaginal douching, number of lifetime sexual partners, and use of illicit drugs) also were significantly associated with the presence of BV . After controlling for the effects of these variables, using a multivariable logistic regression model, chronic maternal stress remained a significant and independent predictor of BV status . Women in the moderate-stress group (third quartile) and high-stress (fourth quartile) group were 2.3 times (95% CI = 1.2-4.3) and 2.2 times (95% CI = 1.1-4.2) more likely to be BV+ than women in the low-stress group (bottom quartile) . CONCLUSIONS: High levels of chronic stress during pregnancy are associated with bacterial vaginosis . The effect of chronic maternal stress is independent of the effects of other established sociodemographic and behavioral risk factors for BV. Microbiol Res, 2001, 156(2), 159 - 68 Screening for trbB- and traG-like sequences by PCR for the detection of conjugative plasmids in bacterial soil isolates; Disque-Kochem C et al.; The transfer regions of different conjugative plasmids show significant similarities in the genetic organization and in the amino acid sequence of some gene products, especially of proteins from the traG or trbB family . These similarities are also evident on the level of the nucleotide sequences . On the basis of conserved DNA regions we designed degenerate PCR primer pairs to detect specifically tra regions within a collection of bacterial clones isolated from an agricultural soil . Most of the potential transfer-proficient indigenous bacterial isolates were able to mobilize a derivative of the nonconjugative IncQ plasmid RSF1010 into recipient strains . With the help of the primers it should be possible to evaluate the genetic potential for horizontal gene transfer carried out by conjugative plasmids. Microbiol Res, 2001, 156(2), 121 - 31 Physiological and genetic characteristics of two bacterial strains utilizing phenoxypropionate and phenoxyacetate herbicides; Muller RH et al.; Two strains, Rhodoferax sp . P230 and Delftia (Comamonas) acidovorans MCI, have previously been shown to carry activities for the degradation of the two enantiomers of (RS)-2-(2,4-dichlorophenoxy-)propionate (dichlorprop) and (RS)-2-(4-chloro-2-methylphenoxy-)propionate (mecoprop) and, in addition, are capable of degrading phenoxyacetate derivatives 2.4-dichlorophenoxyacetate (2,4-D) and 4-chloro-2-methylphenoxyacetate (MCPA) . Metabolism of the herbicides is initiated by alpha-ketoglutarate-dependent dioxygenases for both enantiomers of the phenoxypropionate herbicides and for 2,4-D . These activities were constitutively expressed for both enantiomers of dichlorprop in strain MC1 and for the Renantiomer in strain P230 . Enzyme activities for the complete degradation of phenoxyacetate and phenoxypropionate herbicides were induced during incubation on either of these herbicides . Strain MC1 has about threefold higher activities for the degradation of dichlorprop and for growth on this substrate (mumax = 0.15 h(-1)) than strain P230; the maximum growth rate on 2,4-D amounts to 0.045 h(-1) with strain MC1 . Dichlorprop is utilized faster than mecoprop and the R-enantiomers are cleaved with higher rates than the S-enantiomers . The degradation of the chlorophenolic intermediates seems to proceed via the modified ortho cleavage pathway as indicated by activities of the respective enzymes . The enzymatic results were supported by genetic investigations by which the presence of the genes tfdB (encoding a dichlorophenol hydroxylase), tfdC (encoding a chlorocatechol 1,2-dioxygenase) and tfdD (encoding a chloromuconate cycloisomerase) could be demonstrated in both strains by PCR after application of respective primers . The presence of the tfdA gene (encoding a 2,4-D/alpha-ketoglutarate dioxygenase) was only shown for strain P230 but was lacking in strain MC1 . Sequence analysis of the tfd gene fragments revealed high homology to the degradative genes of other proteobacterial strains degrading chloroaromatic compounds . Strain MC1 carries a plasmid of about 120 kb which apparently harbors herbicide degradative genes as concluded from deletion mutants which have lost 2,4-D{phenoxalkanoate}/alpha-ketoglutarate dioxygenase activities for cleavage of the R- and S-enantiomer, and of 2,4-D . For strain P230, no plasmid could be demonstrated; the activity was stably conserved in this strain during growth under nonselective conditions. Bone Marrow Transplant, 2001 Aug, 28(4), 413 - 5 Successful unrelated BMT in a patient with Kostmann syndrome complicated by pre-transplant pulmonary 'bacterial' abscesses; Toyoda H et al.; Kostmann syndrome, severe congenital neutropenia, is often associated with life-threatening bacterial infections . A 5-year-old girl with Kostmann syndrome developed pulmonary abscesses . She was refractory to granulocyte colony-stimulating factor and antibiotics . She underwent unrelated HLA-matched BMT . Myeloablative conditioning consisted of 12-Gy TBI with lung shielding, antithymocyte globulin, etoposide, and cyclophosphamide . After successful engraftment, the pulmonary abscesses resolved by day 75 post-transplant . Although the option of transplantation is not established in the setting of unrelated HLA-matched BMT in Kostmann syndrome, this case may provide useful information . Furthermore, pre-transplant pulmonary bacterial abscesses may not be a contraindication for BMT in some patients with Kostmann syndrome. Appl Environ Microbiol, 2001 Oct, 67(10), 4896 - 900 Assessing the diversity of marine bacterial beta-glucosidases by capillary electrophoresis zymography; Arrieta JM et al.; We propose a new method for the fast separation and detection of beta-glucosidases in environmental samples . With this approach, beta-glucosidases extracted from bacteria are evidenced by substrate-incorporated capillary electrophoresis (CE zymography) and their kinetic parameters can be determined by repeated injections using different substrate concentrations . Preliminary results obtained with natural bacterial communities from the coastal North Sea suggest that the diversity of beta-glucosidases in the marine environment might be much higher than previously observed. Appl Environ Microbiol, 2001 Oct, 67(10), 4531 - 7 Evidence for the biosynthesis of bryostatins by the bacterial symbiont "Candidatus Endobugula sertula" of the bryozoan Bugula neritina; Davidson SK et al.; The marine bryozoan, Bugula neritina, is the source of the bryostatins, a family of macrocyclic lactones with anticancer activity . Bryostatins have long been suspected to be bacterial products . B . neritina harbors the uncultivated gamma proteobacterial symbiont "Candidatus Endobugula sertula." In this work several lines of evidence are presented that show that the symbiont is the most likely source of bryostatins . Bryostatins are complex polyketides similar to bacterial secondary metabolites synthesized by modular type I polyketide synthases (PKS-I) . PKS-I gene fragments were cloned from DNA extracted from the B . neritina-"E . sertula" association, and then primers specific to one of these clones, KSa, were shown to amplify the KSa gene specifically and universally from total B . neritina DNA . In addition, a KSa RNA probe was shown to bind specifically to the symbiotic bacteria located in the pallial sinus of the larvae of B . neritina and not to B . neritina cells or to other bacteria . Finally, B . neritina colonies grown in the laboratory were treated with antibiotics to reduce the numbers of bacterial symbionts . Decreased symbiont levels resulted in the reduction of the KSa signal as well as the bryostatin content . These data provide evidence that the symbiont E . sertula has the genetic potential to make bryostatins and is necessary in full complement for the host bryozoan to produce normal levels of bryostatins . This study demonstrates that it may be possible to clone bryostatin genes from B . neritina directly and use these to produce bryostatins in heterologous host bacteria. Arch Dis Child, 2001 Oct, 85(4), 346 - 7 Effect of bacterial filters on spirometry measurements; Kamps AW et al.; Lung function measurements with and without a disposable bacterial filter were compared in 60 children . Although statistically significant, the reduction of lung function measurements caused by using bacterial filters was small and clinically irrelevant (2-4% of predicted for forced expiratory volume in one second and vital capacity). EMBO J, 2001 Sep 17, 20(18), 5280 - 9 Bacterial senescence: protein oxidation in non-proliferating cells is dictated by the accuracy of the ribosomes; Ballesteros M et al.; We have investigated the causal factors behind the age-related oxidation of proteins during arrest of cell proliferation . A proteomic approach demonstrated that protein oxidation in non-proliferating cells is observed primarily for proteins being produced in a number of aberrant isoforms . Also, these cells exhibited a reduced translational fidelity as demonstrated by both proteomic analysis and genetic measurements of nonsense suppression . Mutants harboring hyperaccurate ribosomes exhibited a drastically attenuated protein oxidation during growth arrest . In contrast, oxidation was augmented in mutants with error-prone ribosomes . Oxidation increased concomitantly with a reduced rate of translation, indicating that the production of aberrant, and oxidized proteins, is not the result of titration of the co-translational folding machinery . The age-related accumulation of the chaperones, DnaK and GroEL, was drastically attenuated in the hyperaccurate rpsL mutant, demonstrating that the reduced translational fidelity in growth-arrested cells may also be a primary cause for the induction of the heat shock regulon . The data point to an alternative way of approaching the causal factors involved in protein oxidation in eukaryotic G(0) cells. Clin Sci (Lond), 2001 Oct, 101(4), 395 - 402 Interaction between bacterial peptides, neutrophils and goblet cells: a possible mechanism for neutrophil recruitment and goblet cell depletion in colitis; Leiper K et al.; Goblet cell depletion occurs in various forms of colitis, but its mechanism is unknown . We have investigated two linked hypotheses: (i) that bacterial peptides, such as formyl-methionyl-leucyl-phenylalanine (fMLP), interact with epithelial cells inducing the release of chemokines, including interleukin-8 (IL-8), which in turn leads to the recruitment of neutrophils which release mucin secretagogues; (ii) that fMLP acts directly on epithelial cells to cause mucus secretion . Studies were performed to measure the effects of fMLP on the synthesis and secretion of IL-8 and mucus by the goblet cell differentiated colon cancer cell lines HT29-MTX (methotrexate-conditioned HT29 colonic adenocarcinoma cell line) and LS174T, and to assess the effects of neutrophil-derived secretagogues on goblet cell secretion in these cell lines . fMLP (0.1 microM) increased the secretion of IL-8 by 105% (P<0.0001) in HT29-MTX cells and by 401% (P<0.0001) in LS174T cells . fMLP also increased the synthesis and secretion of mucins by these cell lines, with maximal effects of 65% above control values for synthesis (P<0.01) and 73% for secretion (P<0.01) . A dose-related increase (up to 67%; P<0.01) in mucin secretion was demonstrated in HT29-MTX cells in response to incubation with supernatant from activated neutrophils . This effect was largely (83%; P<0.02) inhibited by ICI 200,355, a specific inhibitor of neutrophil elastase . In conclusion, the bacterial peptide fMLP and neutrophil elastase are both potent mucus secretagogues for colon epithelial cells . fMLP also elicits release of the potent neutrophil chemoattractant IL-8 from colon epithelial cells . These findings support the hypothesis that the mucosal inflammation and mucus depletion seen in ulcerative colitis could result from interaction between bacterial peptides and the mucosa. J Biol Chem, 2001 Dec 7, 276(49), 45669 - 76 Epub 2001 Sep 19. Sensitivity of different ecotypes and mutants of Arabidopsis thaliana toward the bacterial elicitor flagellin correlates with the presence of receptor-binding sites; Bauer Z et al.; Flagellin, the main building block of the bacterial flagellum, acts as potent elicitor of defense responses in different plant species . Genetic analysis in Arabidopsis thaliana identified two distinct loci, termed FLS1 and FLS2, that are essential for perception of flagellin-derived elicitors . FLS2 was found to encode a leucine-rich repeat transmembrane receptor-like kinase with similarities to Toll-like receptors involved in the innate immune system of mammals and insects . Here we used a radiolabeled derivative of flg22, a synthetic peptide representing the elicitor-active domain of flagellin, to probe the interaction of flagellin with its receptor in A . thaliana . The high affinity binding site detected in intact cells and membrane preparations exhibited specificity for flagellin-derived peptides with biological activity as agonists or antagonists of the elicitor responses . Specific binding activity was measurable in all ecotypes of A . thaliana that show sensitivity to flagellin but was barely detectable in the flagellin-insensitive ecotype Ws-0 affected in FLS1 . A strongly impaired binding of flagellin was observed also in several independent flagellin-insensitive mutants isolated from the flagellin-sensitive ecotype La-er . In particular, no binding was found in plants carrying a mutation in the LRR domain of FLS2 . These data indicate that the formation of functional receptor-binding sites depends on genes encoded by both loci, FLS1 and FLS2 . The tight correlation between the presence of the binding site and elicitor response provides strong evidence that this binding site acts as the physiological receptor of flagellin. J Protein Chem, 2001 Feb, 20(2), 139 - 44 The effect of inorganic phosphate on the activity of bacterial ribokinase; Maj MC et al.; Ribokinase and adenosine kinase are both members of the PfkB family of carbohydrate kinases . The activity of mammalian adenosine kinase was previously shown to be affected by pentavalent ions (PVI) . We now present evidence that the catalytic activity of E . coli ribokinase is also affected by PVI, increasing both the velocity and affinity of the enzyme for D-ribose . The Km, for ribose decreased from 0.61 mM to 0.21, 0.25, and 0.33 mM in the presence of 20 mM phosphate, arsenate, and vanadate, respectively . The activity of ribokinase was stimulated in a hyperbolic fashion, with the maximum velocity increasing 23-fold, 13-fold, and 11-fold under the same conditions, respectively . Activity was also affected upon the addition of phosphoenolpyruvate, suggesting that phosphorylated metabolites could be involved in enzymatic control . The similar effect of PVI on distantly related enzymes suggests that a common mechanism for activity is shared among PfkB family members. Environ Sci Technol, 2001 Sep 1, 35(17), 3442 - 7 Bacterial growth in distribution systems: effect of assimilable organic carbon and biodegradable dissolved organic carbon; Escobar IC et al.; Two distribution systems, one treating water by ozonation and another treating water by nanofiltration in parallel with lime softening, were monitored for bacterial growth . Both systems kept disinfectant residuals such as chlorine and chloramine in their respective distribution systems . Bacterial growth was assessed by heterotrophic plate counts (HPC) on R2A agar . In the distribution systems fed by ozonated water, HPCs were correlated (R2 = 0.96) using an exponential model with the assimilable organic carbon (AOC) at each sampling site . Also, it was observed that ozonation caused a significant increase in the AOC concentration of the distribution system (over 100% increase) as well as a significant increase in the bacterial counts of the distribution system (average increase over 100%) . The HPCs from the distribution systems fed by nanofiltration in parallel with lime-softening water also displayed an exponential correlation (R2 = 0.73) with an exponential model based on AOC . No significant correlation was found between bacteria growth on R2A agar and BDOC concentrations . Therefore, in agreement with previous work, bacterial growth in the distribution systems was found to correlate with AOC concentrations. Indian J Pediatr, 2001 Aug, 68(8), 737 - 47 Bacterial meningitis in children: critical care needs; Singhi S et al.; Acute bacterial meningitis (ABM) in children is associated with a high rate of acute complications and mortality, particularly in the developing countries . Most of the deaths occur during first 48 hours of hospitalization . Coma, raised intracranial pressure (ICP), seizures, shock have been identified as significant predictors of death and morbidity . This article reviews issues in critical care with reference to our experience of managing 88 children with ABM in PICU . Attention should first be directed toward basic ABCs of life-support . Children with Glasgow Coma Scale (GSC) score < 8 need intubation and supplemental oxygen . Antibiotics should be started, even without LP (contraindicated if focal neuro-deficit, papilledema, or signs of raised ICP) . Raised ICP is present in most of patients; GCS < 8 and high blood pressure are good guides . Mannitol (0.25 gm/Kg) should be used in such patients . If there are signs of (impending) herniation short-term hyperventilation is recommended; prolonged hyperventilation (> 1 hour) must be avoided . Any evidence of poor perfusion, hypovolemia and/or hypotension needs aggressive treatment with normal saline boluses and inotropes, if necessary, to maintain normal blood pressure . Empiric fluid restriction is not justified . Seizures may be controlled with intravenous diazepam or lorazepam . Refractory status epilepticus may be treated with continuous diazepam (0.01-0.06) mg/kg/min) or midazolam infusion . Ventilatory support may be needed early for associated pneumonia, poor respiratory effort and/or coma, and occasionally to reduce work of breathing in shock . Provision of critical care to children with ABM may reduce the mortality significantly as experienced by us. Nucleosides Nucleotides Nucleic Acids, 2001 Apr-Jul, 20(4-7), 977 - 9 Recombinant bacterial cells as efficient biocatalysts for the production of nucleosides; Spoldi E et al.; The preparation of nucleosides as well as their base-modified analogues using purified nucleoside phosphorylase enzymes or, more conveniently, using whole bacterial cells is described . The development of genetically modified strains of Escherichia coli, able to over-produce Uridine-phosphorylase and Purine-nucleoside-phosphorylase in the same cells, improves the specific biocatalytic activity and the consequent industrial scale approach. Mol Cells, 2001 Aug 31, 12(1), 117 - 20 Construction and characterization of a bacterial artificial chromosome library from chili pepper; Yoo EY et al.; A library of the bacterial artificial chromosome (BAC) that consisted of a total of 78,336 clones with an average insert size of 80 kb was constructed from Capsicum annuum, 'CM334', which is resistant to Phytophthora capsici and PVY . Based on a haploid genome size of pepper of 2,702 Mbp/C, the BAC library was estimated to contain approximately three genome equivalents and represented at least 90% of the pepper genome . In order to determine the percentage of BAC clones that contained mitochondrial DNAs, the entire library was screened with probes of chili pepper mitochondrial DNAs . The result showed that only twenty-five clones, which is 0.03% of the total BAC clones, were hybridized to mitochondrial gene probes . This indicates that the library is comprised predominantly of the nuclear sequences . The library was also tested for isolating specific clones by screening with a few known genes from the chili pepper, phytoene synthase gene, and two MADS genes--HpMADS1 and HpMADS3 . The result showed that the three clones for phytoene synthase and the two clones for each MADS gene were positively hybridized to the specific probes . This indicates that the library is highly reliable and represents a resource for initiating map-based cloning and contig mapping in chili pepper. Lett Appl Microbiol, 2001 Oct, 33(4), 296 - 301 A comparison of nucleic acid amplification techniques for the assessment of bacterial viability; Birch L et al.; AIMS: The ability to determine the presence and viability status of bacteria by molecular methods could offer significant advantages to the food, environmental and health sectors, in terms of improved speed and sensitivity of detection . METHODS AND RESULTS: In this study, we have assessed three amplification techniques, PCR, RT-PCR and NASBA, for their ability to detect nucleic acid persistence in an E . coli strain following heat-killing . NASBA offered the greatest sensitivity of the three methods tested . The presence of residual DNA and mRNA could be detected by PCR and NASBA, respectively, for up to 30 h postdeath, by which time cell death had been confirmed by culture methods . Thus a single quantitative measurement based on nucleic acid amplification did not permit unequivocal determination of cell viability . CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The correlation between cell viability and persistence of nucleic acids must be well characterized for a particular analytical situation before molecular techniques can be substituted for traditional culture methods. Antimicrob Agents Chemother, 2001 Oct, 45(10), 2936 - 8 Comparison of once-daily versus twice-daily administration of cefdinir against typical bacterial respiratory tract pathogens; Ross GH et al.; In an in vitro pharmacodynamic model, a twice-daily cefdinir dosing regimen was more effective than a once-daily regimen against common bacterial respiratory pathogens in producing 3-log(10) killing and preventing the occurrence of regrowth at 24 h . Twice-daily administration is likely the more appropriate cefdinir dosing strategy for the treatment of community-acquired pneumonia. Eur J Intern Med, 2001 Sep, 12(5), 454 - 458 Non-bacterial thrombotic (marantic) endocarditis associated with giant-cell arteritis; Hesselink DA et al.; A patient with giant-cell arteritis and non-bacterial thrombotic (marantic) endocarditis of the mitral valve is described . To our knowledge, this is the first case reported . The importance of revising the diagnosis of infective endocarditis when no pathogen can be demonstrated is emphasized. Brain Pathol, 2001 Oct, 11(4), 422 - 31 Expression of death-related proteins in dentate granule cells in human bacterial meningitis; Gerber J et al.; Neuronal apoptosis in the dentate gyrus has been observed in animal models of bacterial meningitis and in humans dying in the course of the disease.To evaluate the mechanisms of neuronal cell death, hippocampal sections of 20 patients dying from bacterial meningitis were investigated by immunohistochemistry using antibodies against the proform of caspase-3 and the active enzyme, bcl-2, bax and p53 . In the dentate granule cell layer, the median density of neurons with an apoptotic morphology was 7.6/mm2 (0-15.6/mm2) . The median density of immunoreactive neurons was 2.3/mm2 (procaspase-3), 0.9/mm2 (activated caspase-3), 1.8/mm2 (bcl-2), 1.1/mm2 (bax) and 0.4/mm2 (p53) . 80% of neurons immunoreactive for active caspase-3 had an apoptotic morphology, whereas only 10% of all procaspase-3 stained neurons showed signs of apoptosis . Apoptotic cell death is present in humans dying in the course of bacterial meningitis in the dentate gyrus of the Formatio hippocampi . Neuronal expression of caspase-3, bcl-2 and bax suggests an involvement of these proteins in neuronal death. Int J Med Microbiol, 2001 Aug, 291(3), 197 - 207 Modification of host cell apoptosis by viral and bacterial pathogens; Muller A et al.; Bacterial and viral pathogens have evolved sophisticated strategies to manipulate the host cell's life span to their own advantage . This is achieved by modifications of the intrinsic apoptosis program at several checkpoints: kinases, caspases and mitochondria . The goal of this review is to give an overview over the molecular mechanisms involved. Clin Orthod Res, 2001 May, 4(2), 112 - 8 Bacterial degradation of composite bonding materials; Donker HJ et al.; The causes of bracket loss during the late phases of orthodontic treatment have hitherto been considered to be of mechanical nature (e.g . trauma, high forces applied) . Recently, it was hypothesized that bacterial degradation of composite resin could be a reason for late bracket failure . This was based on the observation of apparently degraded composite on bracket bases sent to a recycling company, and on a weight loss of 0.1-1% of a limited number of composite resin foils incubated with bacteria . To further elucidate the basis for this hypothesis, we examined brackets, immediately after debonding, for signs of potential decay and tested the ability of selected bacterial species to degrade composites . Out of a total of 1056 brackets collected, 6.2% displayed signs of decay indicative of inclusion of air bubbles, corrosion of the bracket base and discoloration of the composite and comparable to the decay observed on brackets received from the recycling company . Composite discs of two different brands were each incubated with either of five strains representing different bacterial species . The incubation lasted 3 months or until the bacteria had died . There was no significant difference in weight change of the discs incubated with bacteria and control discs incubated without bacteria . Thus, our results do not support that bacterial degradation of composite resin bonding materials is a likely cause of bracket loss. Rev Med Chil, 2001 Jul, 129(7), 719 - 26 {Trends of etiology of acute bacterial meningitis in Chilean children from 1989 to 1998 . Impact of the anti-H influenzae type b vaccine}; Diaz JM et al.; BACKGROUND: Acute bacterial meningitis still has a high mortality and rate of complications . AIM: To assess the impact of anti H influenzae vaccination on the epidemiology of acute bacterial meningitis in Chilean children . MATERIAL AND METHODS: A retrospective study of hospital discharge records of patients with acute bacterial meningitis . Causative agents were studied globally, by hospital and by age group . The changes in etiology from 1989 to 1995 were also assessed . Between 1996 and 1998, only those patients with acute bacterial meningitis caused by H influenzae were recollected . RESULTS: In the period prior to vaccination (1989-1995), 1000 cases were registered . The main causative agents were N meningitidis in 33.8%, H influenzae type b in 21.9% and S pneumoniae in 15.4% . The incidence of H influenzae decreased in the period from 36.4 to 9.9% (p < 0.001) and the incidence of N meningitidis increased from 22.9 to 52.1% (p < 0.001) . The incidence of S pneumoniae did not change significantly . H influenzae predominated in children between 4 and 24 months of age and N meningitidis predominated in children over 25 months of age . In the period after the introduction of vaccination (1995-1998), there was a further decrease in the incidence of H influenzae from 10 to 2% (p < 0.001) . Until 1997, there was a considerable increase in the incidence of N meningitidis, specially in children over 25 months of age . It declined in 1998 to 38% . CONCLUSIONS: There was a reduction in the incidence of acute bacterial meningitis caused by H influenzae prior to the introduction of the vaccine against H influenzae type b . The decrease was more pronounced after the introduction of the vaccine. J Struct Biol, 2001 May-Jun, 134(2-3), 219 - 31 Fold predictions for bacterial genomes; Pawlowski K et al.; Fold assignments for newly sequenced genomes belong to the most important and interesting applications of the booming field of protein structure prediction . We present a brief survey and a discussion of such assignments completed to date, using as an example several fold assignment projects for proteins from the Escherichia coli genome . This review focuses on steps that are necessary to go beyond the simple assignment projects and into the development of tools extending our understanding of functions of proteins in newly sequenced genomes . This paper also discusses several problems seldom addressed in the literature, such as the problem of domain prediction and complementary predictions (e.g., transmembrane regions and flexible regions) and cross-correlation of predictions from different servers . The influence of sequence and structure database growth on prediction success is also addressed . Finally, we discuss the perspectives of the field in the context of massive sequence and structure determination projects, as well as the development of novel prediction methods . Zh Mikrobiol Epidemiol Immunobiol, 2001 May-Jun, (3), 46 - 9 {Immunoprophylaxis of acute respiratory diseases with bacterial multicomponent vaccine VP-4 in children at preschool institutions}; Egorova NB et al.; The prophylactic action of polycomponent vaccine B{symbol: see text}-4, prepared from the antigens of opportunistic bacteria, on morbidity rate in acute respiratory diseases (ARD) of bacterial and mixed (bacterial and viral) etiology in 121 children aged 2-5 years, attending pre-school institutions was evaluated . For comparison, a group of 118 children of the same age from the same institutions was formed . The vaccine was introduced after the schedule consisting of 3 intranasal and 6-9 oral administrations made at intervals of 3-4 days . The duration of the course of immunization was 26 +/- 4 days . The prophylactic effect of B{symbol: see text}-4 on ARD morbidity was evaluated by the number of ARD cases and their duration per child . The prophylactic effect of B{symbol: see text}-4 on ARD morbidity lasted 14 months (the term of observation) after immunization and was manifested by a decrease in the number and duration of ARD cases after administration of the preparation, also in a group of highly susceptible children. Water Res, 2001 Oct, 35(14), 3490 - 5 Use of bacterial biosensors to interpret the toxicity and mixture toxicity of herbicides in freshwater; Strachan G et al.; The dose response relationship between seven commonly used herbicides and four luminescence-based bacterial biosensors was characterised . As herbicide concentration increased the light emitted by the test organism declined in a concentration dependent manner . These dose responses were used to compare the predicted vs . observed response of a biosensor in the presence of multiple contaminants . For the majority of herbicide interactions, the relationship was not additive but primarily antagonistic and sometimes synergistic . These biosensors provide a sensitive test and are able to screen a large volume and wide range of samples with relative rapidity and ease of interpretation . In this study biosensor technology has been successfully applied to interpret the interactive effects of herbicides in freshwater environments. Infection, 2001 Aug, 29(4), 209 - 12 Predictive value of serum and cerebrospinal fluid procalcitonin levels for the diagnosis of bacterial meningitis; Jereb M et al.; BACKGROUND: The value of serum and cerebrospinaL fluid (CSF) procalcitonin for differentiating between acute bacterial and viral meningitis was assessed and compared to other parameters which are usually used in clinical practice . PATIENTS: 45 adult patients (20 with bacterial and 25 with tick-borne encephalitis, TBE) were included in this prospective study . RESULTS: The median serum procalcitonin Level in patients with bacterial meningitis was 6.45 ng/ml (range 0.25-43.76 ng/ml) and in the group with viral meningitis 0.27 ng/ml (range 0.05-0.44 ng/ml) . 11 patients with bacterial meningitis had an elevated procalcitonin concentration not only in serum, but also in CSF . A serum procalcitonin Level > 0.5 ng/ml had a positive predictive value for bacterial meningitis of 100% and a negative predictive value of 93%, while corresponding values for CSF procalcitonin were 100% and 74%, respectively . CONCLUSION: Serum and CSF procalcitonin concentrations > 0.5 ng/ml appear to be a reliable indicator of bacterial central nervous system (CNS) infection, with maximal positive predictive values and high negative predictive values. J Mol Microbiol Biotechnol, 2001 Oct, 3(4), 557 - 62 Development of a bacterial screen for novel hypoxanthine-guanine phosphoribosyltransferase substrates; Shivashankar K et al.; The lack of de novo purine biosynthesis in many parasitic protozoans makes the enzymes in the salvage of purines attractive chemotherapeutic targets . Hypoxanthine-guanine phosphoribosyltransferase (HGPRT) is a key enzyme for purine salvage and bacterial complementation screens for HGPRT inhibitors are known . The low KMS for purine bases makes purine analogs unattractive as competitive inhibitors for this enzyme . Despite the availability of many crystal structures of HGPRTs, it is only recently that selective inhibitors of the enzyme have been developed . Therefore, novel purine analogs which act as substrates for the HGPRT reaction and thereby inhibit downstream enzymes or get incorporated into the nucleotide pool are an attractive altenative for drug design . We have used a combination of two E . coli strains Sphi606 (ara, deltapro-gpt-lac, thi, hpt) and Sphi609 (ara, deltapro-gpt-lac, thi, hpt, pup, purH,J, strA) to identify inhibitors and substrates of HGPRT . E . coli Sphi609 is deficient in both de novo synthesis as well as salvage enzymes of purine nucleotide synthesis, while E . coli Sphi606 is deficient in salvage enzymes only . Hence, expression of functional HGPRTs in E . coli Sphi606 grown in minimal medium makes it susceptible to HGPRT substrates, which inhibit downstream processes . Growth of E . coli Sphi609 in minimal medium can be made conditional for the expression of a functional HGPRT and this growth would be susceptible to both HGPRT substrate analogs and inhibitors . A substance that strictly acts as an inhibitor will affect growth of transformed E . coli Sphi609 only . For this purpose, we compared the human and P . falciparum enzymes with known HGPRT substrate analogs . Our data with 6-mercaptopurine, 6-thioguanine and allopurinol show that these compounds act by being substrates for HGPRT . Our results with allopurinol suggest that it is a better substrate for P . falciparum HGXPRT than the human enzyme . Therefore, species-specific substrates can be tested out successfully in E . coli Sphi606 . The formation of products from substrates like allopurinol lacking a labile proton at N7 raises the possibility that the deprotonation of substrates might occur at N9 rather than at N7 or a purine anion might be the true substrate for the reaction. Annu Rev Microbiol, 2001, 55, 407 - 35 Interaction of bacterial pathogens with polarized epithelium; Kazmierczak BI et al.; Many pathogens must surmount an epithelial cell barrier in order to establish an infection . While much has been learned about the interaction of bacterial pathogens with cultured epithelial cells, the influence of cell polarity on these events has only recently been appreciated . This review outlines bacterial-host epithelial cell interactions in the context of the distinct apical and basolateral surfaces of the polarized epithelium that lines the lumens of our organs. J Bacteriol, 2001 Oct, 183(19), 5506 - 12 Autophosphorylation of a bacterial serine/threonine kinase, AfsK, is inhibited by KbpA, an AfsK-binding protein; Umeyama T et al.; A protein serine/threonine kinase, AfsK, and its target protein AfsR globally control physiological and morphological differentiation in the bacterial genus Streptomyces . A protein (KbpA) of 252 amino acids encoded by an open reading frame in a region upstream of afsK in Streptomyces coelicolor A3(2) was identified as an AfsK-interacting protein . The interaction site of AfsK was in the N-terminal portion containing the kinase catalytic domain . KbpA bound a nonphosphorylated form of AfsK and inhibited its autophosphorylation at serine and threonine residues . KbpA in the reaction mixture containing AfsK and AfsR also inhibited the phosphorylation of AfsR by AfsK, presumably because KbpA inhibited the conversion from the inactive, nonphosphorylated form of AfsK to the active, phosphorylated form . kbpA was transcribed throughout growth, and the transcription was enhanced when production of actinorhodin had already started . KbpA thus appeared to play an inhibitory role in a negative feedback system in the AfsK-AfsR regulatory pathway . Consistent with these in vitro observations, kbpA served as a repressor for actinorhodin production in S . coelicolor A3(2); disruption of kbpA greatly enhanced actinorhodin production, and overexpression of kbpA reduced the production. Comp Biochem Physiol C Toxicol Pharmacol, 2001 Sep, 130(1), 107 - 17 Eicosanoids act in nodulation reactions to bacterial infections in newly emerged adult honey bees, Apis mellifera, but not in older foragers; Bedick JC et al.; Nodulation is the first, and qualitatively predominant, cellular defense reaction to bacterial infections in insects . We tested the hypothesis that eicosanoids also mediate nodulation reactions to bacterial challenge in adults of a social insect, the honey bee, Apis mellifera . Treating newly-emerged experimental bees with the eicosanoid biosynthesis inhibitor, dexamethasone, impaired nodulation reactions to bacterial infections, and the influence of dexamethasone was reversed by treating infected insects with arachidonic acid, an eicosanoid precursor . Several other eicosanoid biosynthesis inhibitors, including the cyclooxygenase inhibitor, indomethacin, and the dual cyclooxygenase/lipoxygenase inhibitor, phenidone, also impaired the ability of experimental honeybees to form nodules in reaction to bacterial challenge . The influence of phenidone on nodulation was expressed in a dose-dependent manner . However, in experiments with older honey bees foragers, similar bacterial challenge did not evoke nodulation reactions . We infer from our results that while eicosanoids mediate cellular immune responses to bacterial infections in newly emerged honey bees, and more broadly, in most insect species, nodulation reactions to bacterial challenge probably do not occur in all phases of insect life cycles. Microgravity Sci Technol, 1999, 12(2), 74 - 7 Effects of space flight and mixing on bacterial growth in low volume cultures; Kacena MA et al.; Previous investigations have shown that liquid suspension bacterial cultures grow to higher cell concentrations in spaceflight than on Earth . None of these studies included ground-control experiments designed to evaluate the fluid effects potentially responsible for the reported increases . Therefore, the emphasis of this research was to both confirm differences in final cell concentration between 1g and microgravity cultures, and to examine the effects of mixing as a partial explanation for this difference . Flight experiments were performed in the Fluid Processing Apparatus (FPA), aboard Space Shuttle Missions STS-63 and STS-69, with simultaneous 1g static and agitated controls . Additional static 1g, agitated, and clino-rotated controls were performed in 9-ml culture tubes . This research revealed that both E . coli and B . subtilis samples cultured in space flight grew to higher final cell densities (120-345% increase) than simultaneous static 1g controls . The final cell concentration of E . coli cells cultured under agitation was 43% higher than in static 1g cultures and was 102% higher with clino-rotation . However, for B . subtilis cultures grown while being agitated on a shaker or clino-rotated, the final cell concentrations were nearly identical to those of the simultaneous static 1g controls . Therefore, these data suggest that the unique fluid quiescence in the microgravity environment (lack of sedimentation, creating unique transfer of nutrients and waste products), was responsible for the enhanced bacterial proliferation reported in this and other studies. Adv Space Res, 1983, 3(8), 95 - 104 Effect of heavy ions on bacterial spores; Takahashi T et al.; Inactivation of B . subtilis spores has been studied using accelerated He, C, N, O and Ne ions . The energy dependence of the inactivation cross sections for heavy ions was very weak and the mean cross sections for carbon ions (0.6-4.7 MeV/amu), nitrogen ions (0.6-4.1 MeV/amu), oxygen ions (0.8-1.1 MeV/amu), and neon ions (2.2-3.7 MeV/amu) were found to be about 0.22, 0.23, 0.26, and 0.33 micrometer2 , respectively . Analysis was carried out along lines similar to Katz's target theory but the parameters were chosen so that they have an experimental basis. Adv Space Res, 1983, 3(8), 73 - 8 Physical determinants of radiation sensitivity in bacterial spores; Powers EL; Several factors modifying radiation sensitivity in dry bacterial spores are described and discussed . Vacuum inducing the loss of critical structural water, very low dose rates of radiation from which the cell may recover, radiations of high linear energy transfer, and the action of temperature over long periods of time on previously irradiated cells are recognized from extensive laboratory work as important in determining survival of spores exposed to low radiation doses at low temperatures for long periods of time . Some extensions of laboratory work are proposed. Adv Space Res, 1983, 3(8), 105 - 12 Oxygen effect, hydrogen peroxide yields, and time scale of interaction of potentially damaging species in electron pulse irradiated bacterial spores; Tallentire A; A given integrated radiation dose delivered from a LINAC as a train of pulses (50/s), characteristically of 0.1 to 5 microseconds pulse length with dose rates within the pulse between 0.38 and 38 krads/microsecond, inactivates bacterial spores in water suspension more effectively than the same dose given as Co60 gamma rays . This enhancement of radiation damage occurs both in the presence and in the absence of oxygen and is not explained by either pulse dose rate or pulse length alone, but is monotonically related to the product of these pulse parameters, pulse dose . The enhancement appears to result from the interaction, within individual spores, of free radical species of average lifetime of about 2-5 microseconds . The time scale over which these species operate suggests that they are freely diffusable . Prevention, in part, of their damaging effect by the presence of selective scavenging agents is evidence that OH radicals are involved . Measurements of H2O2 yield for irradiation conditions that show a gradation of enhancement of damage correlate strongly with the extent of damage observed. Science, 1994 Dec 23, 266, 1973 - 5 The production of 34S-depleted sulfide during bacterial disproportionation of elemental sulfur; Canfield DE et al.; Bacteria that disproportionate elemental sulfur fractionate sulfur isotopes such that sulfate is enriched in sulfur-34 by 12.6 to 15.3 per mil and sulfide is depleted in sulfur-34 by 7.3 to 8.6 per mil . Through a repeated cycle of sulfide oxidation to S0 and subsequent disproportionation, these bacteria can deplete sedimentary sulfides in sulfur-34 . A prediction, borne out by observation, is that more extensive sulfide oxidation will lead to sulfides that are more depleted in sulfur-34 . Thus, the oxidative part of the sulfur cycle creates circumstances by which sulfides become more depleted in sulfur-34 than would be possible with sulfate-reducing bacteria alone. Genome Biol . 2000;1(6):REVIEWS0006 . Epub 2000 Dec 08. Evaluating genome dynamics: the constraints on rearrangements within bacterial genomes; Hughes D; Inversions and translocations distinguish the genomes of closely related bacterial species, but most of these rearrangements preserve the relationship between the rearranged fragments and the axis of chromosome replication . Within species, such rearrangements are found less frequently, except in the case of clinical isolates of human pathogens, where rearrangements are very frequent. FEBS Lett, 2001 Aug 31, 504(3), 104 - 11 Carbohydrate transporters of the bacterial phosphoenolpyruvate: sugar phosphotransferase system (PTS); Siebold C et al.; The glucose transporter of Escherichia coli couples translocation with phosphorylation of glucose . The IICB(Glc) subunit spans the membrane eight times . Split, circularly permuted and cyclized forms of IICB(Glc) are described . The split variant was 30 times more active when the two proteins were encoded by a dicistronic mRNA than by two genes . The stability and activity of circularly permuted forms was improved when they were expressed as fusion proteins with alkaline phosphatase . Cyclized IICB(Glc) and IIA(Glc) were produced in vivo by RecA intein-mediated trans-splicing . Purified, cyclized IIA(Glc) and IICB(Glc) had 100% and 30% of wild-type glucose phosphotransferase activity, respectively . Cyclized IIA(Glc) displayed increased stability against temperature and GuHCl-induced unfolding. Mol Microbiol, 2001 Aug, 41(3), 645 - 52 Ectopic transposition of a group II intron in natural bacterial populations; Munoz E et al.; Self-splicing group II introns are thought to be the evolutionary progenitors of eukaryotic spliceosomal introns . The invasion of novel (ectopic) sites by group II introns is considered to be a key mechanism by which spliceosomal introns may have become widely dispersed . However, the dynamics of these events in populations are unknown . In bacteria, only two group II introns have been shown to splice and to be mobile in vivo . One of these introns, RmInt1 from Sinorhizobium meliloti, which encodes a protein with no endonuclease domain, has been shown to invade the ectopic oxi1 site independently of recombinase . In this study, we analysed ectopic transposition of the RmInt1 intron in a natural population of S . meliloti . We characterized S . meliloti isolates by polymerase chain reaction amplification of a gene, dapB, which is found only on the pRmeGR4b plasmid diagnostic of GR4-type strains . The diversity within this specific field population of bacteria was analysed by restriction fragment length polymorphism using ISRm2011-2 (homing site of RmInt1) and RmInt1 as probes . We found that ectopic transposition of RmInt1 to the oxi1 site occurred in this natural bacterial population . This ectopic transposition was also the most frequent genetic event observed . This work provides further evidence that the ectopic transposition of group II introns is an important mechanism for their spread in natural bacterial populations. Eur J Biochem, 2001 Sep, 268(17), 4794 - 801 Bacterial expression and conformational analysis of a chemosensory protein from Schistocerca gregaria; Picone D et al.; Chemosensory proteins (CSPs) are a class of small, soluble proteins present at high concentrations in chemosensory organs of different insect species . Several pieces of evidence suggest their involvement in carrying chemical messages from the environment to chemosensory receptors . However, a structural description of the mechanism of delivery has not been reported . In order to provide the first detailed conformational characterization of these molecules, we cloned a specific isoform (CSP-sg4) from Schistocerca gregaria and expressed it in Escherichia coli . The product was obtained with yields of more than 20 mg per L of culture, all in its soluble form . The recombinant protein was identical to the native one with respect to pairing of the disulfide bridges, aggregative state and secondary structure elements . Structural investigations revealed a significantly stable polypeptide with respect to variations in temperature and acidity . CD analysis, preliminary NMR data and secondary structure prediction pointed to a correctly folded structure where helical regions and loops are alternated in a similar fashion as that observed for other classes of odorant- and pheromone-binding proteins presenting no sequence similarity to CSPs. Shock, 2001 Sep, 16(3), 183 - 8 PGE2 suppresses intestinal T cell function in thermal injury: a cause of enhanced bacterial translocation; Choudhry MA et al.; Increased gut bacterial translocation in burn and trauma patients has been demonstrated in a number of previous studies, however, the mechanism for such an increased gut bacterial translocation in injured patients remains poorly understood . Utilizing a rat model of burn injury, in the present study we examined the role of intestinal immune defense by analyzing the T cell functions . We investigated if intestinal T cells dysfunction contributes to bacterial translocation after burn injury . Also our study determined if burn-mediated alterations in intestinal T cell functions are related to enhanced release of PGE2 . Finally, we examined whether or not burn-related alterations in intestinal T cell function are due to inappropriate activation of signaling molecule P59fyn, which is required for T cell activation and proliferation . The results presented here showed an increase in gut bacterial accumulation in mesenteric lymph nodes after thermal injury . This was accompanied by a decrease in the intestinal T cell proliferative responses . Furthermore, the treatments of burn-injured animals with PGE2 synthesis blocker (indomethacin or NS398) prevented both the decrease in intestinal T cell proliferation and enhanced bacterial translocation . Finally, our data suggested that the inhibition of intestinal T cell proliferation could result via PGE2-mediated down-regulation of the T cell activation-signaling molecule P59fyn . These findings support a role of T cell-mediated immune defense against bacterial translocation in burn injury. Klin Lab Diagn, 2001 Jul, (7), 15 - 9 {Diagnostic significance of the cerebrospinal fluid protein spectrum in bacterial and viral meningitis in children}; Alekseeva LA et al.; Protein spectrum of the cerebrospinal fluid (CSF) was studied by polyacrylamide gel disk electrophoresis in 119 children with bacterial and 17 with viral meningitis of different origin over the course of disease . Liquor proteinograms of patients were compared with those of 37 children without meningitis . Protein spectra of the CSF were shifted in the patients . These shifts can serve as differential diagnostic markers: types 1-2 and 2-2 haptoglobins and beta-lipoprotein appears in the blood during the acute period of disease . More pronounced and stubborn changes in the CSF protein spectrum in pneumococcal and hemophilic meningitis correlated with a more severe clinical course and formation of residual neurological consequences . Progress of dysproteinemic shifts in the protein spectrum of the CSF was characterized by a many-fold increase in the concentrations of high-molecular-weight proteins and decreased relative content of liquor-specific fractions, indicating sharp impairment of the blood-brain barrier permeability and formation of brain edema . Disk electrophoresis method is characterized by high resolution, informative value, requires low amounts of CSF for analysis, and is recommended as an accessory diagnostic test for infectious hospitals. Obstet Gynecol, 2001 Sep, 98(3), 476 - 80 Interleukin-10 administration and bacterial endotoxin-induced preterm birth in a rat model; Terrone DA et al.; OBJECTIVE: To determine whether intra-uterine infusion of interleukin-10 prevents preterm delivery in rats treated with endotoxin . METHODS: Pregnant rats underwent implantation of uterine catheters and were randomly assigned to receive intrauterine infusion of either normal saline, 50 microg lipopolysaccharide endotoxin, or 50 microg lipopolysaccharide with 500 ng interleukin-10 administered either concurrently or 24 hours later . The interval from infusion to delivery for each group was recorded, along with the number of live born pups and their birth weight . We calculated that to obtain a power of 80%, assuming a 24-hour difference in the treatment to delivery times between the test and control subjects, at least six animals would be needed in each group . RESULTS: In females receiving lipopolysaccharide (50 microg) alone, the interval to delivery (P <.05), live birth rate (P <.05), and pup weight (P <.001) were reduced compared with the saline-infused controls . In contrast, females receiving interleukin-10 at the time of the endotoxin challenge or 24 hours after delivered at term with no difference in litter size or live birth weight compared with the controls . CONCLUSION: Animals treated with both lipopolysaccharide and interleukin-10, administered concurrently or 24 hours after the endotoxin challenge, delivered normal weight pups at term with a similar litter size as the saline-infused controls . Interleukin-10 appears to be effective in preventing endotoxin-induced preterm birth and fetal wastage in pregnant rats. Clin Infect Dis, 2001 Oct 1, 33(7), 1022 - 7 Epub 2001 Sep 05. Gardnerella vaginalis isolated from patients with bacterial vaginosis and from patients with healthy vaginal ecosystems; Aroutcheva AA et al.; The differences in the phenotype and genotype of Gardnerella vaginalis isolates from patients with bacterial vaginosis (BV) and from patients without BV are unknown . In our study, 43 isolates of G . vaginalis were examined for biotype (hippurate hydrolysis, lipase, and beta-galactosidase activity), sensitivity to metronidazole, and genotype . Of the 117 women visiting the gynecology clinic at Rush-Presbyterian-St . Luke's Medical Center who were included in the study, 27.4% were found to have BV . G . vaginalis was found in samples from 87.5% of women with BV, from 34.0% of women with intermediate BV, and from 26.4% of women with healthy vaginal ecosystems . Among patients with G . vaginalis, biotypes 7 and 8 were isolated from 32% and 20% of patients, respectively . Biotype 5 was predominantly associated with a healthy vaginal ecosystem (P=.0004) . Biotypes 5 and 7 were the most resistant to metronidazole . No specific phenotype or genotype of G . vaginalis causes BV. Clin Infect Dis, 2001 Oct 1, 33(7), 969 - 75 Epub 2001 Aug 22. Community-acquired bacterial meningitis in adults: categorization of causes and timing of death; McMillan DA et al.; The relationship between cause and timing of death in 294 adults who had been hospitalized with community-acquired bacterial meningitis was investigated . For 74 patients with community-acquired bacterial meningitis who died during hospitalization, the underlying and immediate causes of death were identified according to the criteria of the World Health Organization and National Center for Health Statistics . Patients were classified into 3 groups: category I, in which meningitis was the underlying and immediate cause of death (59% of patients; median duration of survival, 5 days); category II, in which meningitis was the underlying but not immediate cause of death (18%; median duration of survival, 10 days); and category III, in which meningitis was neither the underlying nor immediate cause of death (23%; median duration of survival, 32 days) . In a substantial proportion of adults hospitalized with community-acquired bacterial meningitis, meningitis was neither the immediate nor the underlying cause of death . A 14-day survival end point discriminated between deaths attributable to meningitis and those with another cause. Appl Environ Microbiol, 2001 Sep, 67(9), 3795 - 801 Light conditions affect the measurement of oceanic bacterial production via leucine uptake; Moran XA et al.; The effect of irradiance in the range of 400 to 700 nm or photosynthetically active radiation (PAR) on bacterial heterotrophic production estimated by the incorporation of 3H-leucine (referred to herein as Leu) was investigated in the northwestern Mediterranean Sea and in a coastal North Atlantic site, with Leu uptake rates ranging over 3 orders of magnitude . We performed in situ incubations under natural irradiance levels of Mediterranean samples taken from five depths around solar noon and compared them to incubations in the dark . In two of the three stations large differences were found between light and dark uptake rates for the surface most samples, with dark values being on average 133 and 109% higher than in situ ones . Data obtained in coastal North Atlantic waters confirmed that dark enclosure may increase Leu uptake rates more than threefold . To explain these differences, on-board experiments of Leu uptake versus irradiance were performed with Mediterranean samples from depths of 5 and 40 m . Incubations under a gradient of 12 to 1,731 micromol of photons m(-2) x s(-1) evidenced a significant increase in incorporation rates with increasing PAR in most of the experiments, with dark-incubated samples departing from this pattern . These results were not attributed to inhibition of Leu uptake in the light but to enhanced bacterial response when transferred to dark conditions . The ratio of dark to light uptake rates increased as dissolved inorganic nitrogen concentrations decreased, suggesting that bacterial nutrient deficiency was overcome by some process occurring only in the dark bottles. Trends Immunol, 2001 Sep, 22(9), 497 - 501 Bacterial superantigens: provocateurs of gut dysfunction and inflammation? McKay DM. Inflammatory bowel diseases (IBDs) are chronic debilitating conditions, which impair the patient's quality of life significantly . Among them, Crohn's disease and ulcerative colitis are idiopathic disorders for which an infective etiology has long been sought . Here, we present an opinion in support of the hypothesis that bacterial superantigens can participate in the initiation, exaggeration or reactivation of enteric inflammatory disease, at least in some patients . Although the identification of a specific pathogen responsible for IBD remains a worthy pursuit, an awareness of the response to bacterial products per se will be of value in providing a comprehensive understanding of enteric pathophysiological mechanisms and their potential role in IBDs. J Am Chem Soc, 2001 Sep 5, 123(35), 8550 - 63 Modeling the bacterial photosynthetic reaction center . 4 . The structural, electrochemical, and hydrogen-bonding properties of 22 mutants of Rhodobacter sphaeroides; Hughes JM et al.; Site-directed mutagenesis has been employed by a number of groups to produce mutants of bacterial photosynthetic reaction centers, with the aim of tuning their operation by modifying hydrogen-bond patterns in the close vicinity of the "special pair" of bacteriochlorophylls P identical with P(L)P(M) . Direct X-ray structural measurements of the consequences of mutation are rare . Attention has mostly focused on effects on properties such as carbonyl stretching frequencies and midpoint potentials to infer indirectly the induced structural modifications . In this work, the structures of 22 mutants of Rhodobacter sphaeroides have been calculated using a mixed quantum-mechanical molecular-mechanical method by modifying the known structure of the wild type . We determine (i) the orientation of the 2a-acetyl groups in the wild type, FY(M197), and FH(M197) series mutants of the neutral and oxidized reaction center, (ii) the structure of the FY(M197) mutant and possible water penetration near the special pair, (iii) that significant protein chain distortions are required to assemble some M160 series mutants (LS(M160), LN(M160), LQ(M160), and LH(M160) are considered), (iv) that there is competition for hydrogen-bonding between the 9-keto and 10a-ester groups for the introduced histidine in LH(L131) mutants, (v) that the observed midpoint potential of P for HL(M202) heterodimer mutants, including one involving also LH(M160), can be correlated with the change of electrostatic potential experienced at P(L), (vi) that hydrogen-bond cleavage may sometimes be induced by oxidation of the special pair, (vii) that the OH group of tyrosine M210 points away from P(M), and (viii) that competitive hydrogen-bonding effects determine the change in properties of NL(L166) and NH(L166) mutants . A new technique is introduced for the determination of ionization energies at the Koopmans level from QM/MM calculations, and protein-induced Stark effects on vibrational frequencies are considered. Appl Microbiol Biotechnol, 2001 Jun, 55(6), 673 - 9 Bacterial cellulose production under oxygen-enriched air at different fructose concentrations in a 50-liter, internal-loop airlift reactor; Chao Y et al.; Bacterial cellulose (BC) production by Acetobacter xylinum subsp . sucrofermentans BPR2001 was carried out in a 50-1 internal-loop airlift reactor in air at an initial fructose concentration of 40 g/l . The BC production rate was 0.059 g/l per h . When oxygen-enriched air was supplied instead of air, the BC production rate increased to 0.093 g/l per h, and the BC yield was enhanced from 11% in air to 18% . When the initial fructose concentrations were varied from 30 to 70 g/l, the highest BC yield (35%) the highest production rate (0.22 g/l x per h), and the highest concentration of BC produced (10.4 g/l) were observed at 60-70 g/l fructose . From the carbon mass balance calculated at the final stage of cultivation, it was observed that enhanced BC production was reflected as a decrease in both CO2 evolution and the concentration of other unknown substances, suggesting the efficient utilization of energy for BC synthesis despite O2 limitation. Eur J Epidemiol, 2001, 17(1), 89 - 95 Risk factors for and relationship between bacterial vaginosis and cervicitis in a high risk population for cervicitis in Southern Iran; Keshavarz H et al.; In 1990, a study of the reproductive habits and cervical pathology in women of the Qashgha'i nomadic tribe, resulted in a high prevalence of cervicitis . This led us to explore the likely infectious agents responsible for a such high prevalence; to assess the difference in cervicitis rates between nomadic and non-nomadic populations in the same area; and to determine the risk factors for and the relationship between cervicitis and bacterial vaginosis (BV) . In 1996-1997 a study was carried out of 839 married women of the Qashgha'i, 274 of the Lor nomadic tribes, and 388 non-nomadic urban women . A gynaecological examination, Pap smear and vaginal secretion for assessing BV by gram staining were performed . Data was analysed by logistic regression . Backward stepwise regression was used to assess multivariable effects on risk of cervicitis . The prevalence of cervicitis was 88% in the Qashgha'i, 85% in the Lor and 71% in the urban population . In the multivariate backward stepwise regression analysis for predictors of cervicitis in the Qashgha'i, significant factors were decreased association with those over 40 (p < 0.004) and postpartum bleeding . In the Lor tribe the predictors were an increased risk after more than four pregnancies (p < 0.01) and the use of contraception . Among the urban population, the risk of cervicitis was increased with the use of oral contraceptive previously (p < 0.03) or currently (p < 0.01) . BV was strongly associated with cervicitis, with a relatively high attributable risk . Both sexual and childbirth exposures may be associated with cervicitis in these populations. Stat Med, 2001 Sep 15-30, 20(17-18), 2561 - 71 Statistical considerations of FDA and CPMP rules for the investigation of new anti-bacterial products; Rohmel J; In this paper I investigate statistical properties of some guidance given by the FDA and by the CPMP on the planning, conduct and analysis of clinical trials with new anti-bacterial substances using an active control design . It is demonstrated that the non-inferiority margin proposed by the FDA has some undesirable features, and that the CPMP guidance may need further interpretation with respect to a statement that the non-inferiority margin may be smaller than 10 per cent for response rates >90 per cent . A new margin is proposed that combines the desirable properties from both the FDA and the CPMP guidance . It is also shown that the approximate unconditional tests that are in use in such trials are quite unreliable with respect to preserving the nominal type I error . Unconditional exact tests are presented as a remedy for this issue . J Endotoxin Res, 2000, 6(6), 489 - 91 MD-2 binds to bacterial lipopolysaccharide; Viriyakosol S et al.; Many LPS binding proteins have been described, but the exact nature of the LPS receptors that signal cells remains unclear . MD-2 is a molecule that is found in association with Toll-like receptor 4, which has been shown to be a receptor for LPS . We have produced human MD-2 in baculovirus and tested it for LPS binding . MD-2 binds the lipid A region of LPS without the need for LPS binding protein . These data suggest that MD-2 may be binding LPS as part of the TLR4 receptor complex. J Endotoxin Res, 2000, 6(4), 313 - 9 Bacterial endotoxin (lipopolysaccharide) stimulates the rate of iron oxidation; Roth RI et al.; Bacterial endotoxin (lipopolysaccharide) has affinity for a number of cations, including iron . Previous investigations have demonstrated that lipopolysaccharide can affect the oxidation rate of iron; heme-bound ferrous iron in hemoglobin is oxidized to ferric iron when hemoglobin binds lipopolysaccharide . In the present study, we directly examined the interaction between lipopolysaccharide and iron . Lipopolysaccharide caused a concentration-dependent increase in the rate of iron oxidation, with up to a 23-fold increase in oxidation in the presence of 200 microg/ml Escherichia coli lipopolysaccharide . This effect was seen both with several carbohydrate-rich smooth lipopolysaccharides and also with carbohydrate-poor rough lipopolysaccharide . Extensively deacylated rough lipopolysaccharide had no effect, suggesting a role of the fatty acid components of lipopolysaccharide in this process . Purified lipid A produced inconsistent results: some preparations stimulated iron oxidation and others did not . A series of sugars, starches and a preparation of purified O-chain polysaccharide (the carbohydrate portion of the lipopolysaccharide macro-molecule) had no effect on the rate of iron oxidation, whereas phospholipid-enriched brain tissue extracts (similar to the lipid A component of lipopolysaccharide) stimulated oxidation . We conclude that the lipid moiety of bacterial lipopolysaccharide is responsible for the stimulation of iron oxidation . This process may contribute to the ability of lipopolysaccharide to cause oxidation of heme-bound iron in hemoglobin. Anal Biochem, 2001 Sep 1, 296(1), 73 - 82 Quantification of bacterial polysaccharides by the purpald assay: measurement of periodate-generated formaldehyde from glycol in the repeating unit; Lee CH et al.; We have adapted the purpald assay for measurement of bacterial polysaccharides (PS) containing substituted and/or unsubstituted glycol (SG or UG) in residues such as glycerol, ribitol, arabinitol, furanosyl galactose, and sialic acid . For the purpald assay of UG-containing PS, 50 microL of PS samples was consecutively reacted with 50 microL of 16 mM NaIO4 for 20 min, 50 microL of 136 mM purpald reagent in 2 N NaOH for 20 min, and 50 microL of 64 mM NaIO4 for 20 min in a 96-well tissue culture plate followed by a measurement of absorbance at 550 nm with a plate reader . For SG-containing PS, conversion of SG to UG with 25 micro;L of 0.3 N NaOH, 1 h at room temperature for de-O-acetylation followed by 25 microL of 0.6 M H2SO4, 1 h at 80 degrees C for acid hydrolysis of PS precedes the periodate treatment in the purpald assay . The concentration of the samples can be calculated from the sample absorbance and the reference standard curve constructed from the reference concentrations of the same PS (well-characterized) and their corresponding absorbance values assayed in the same plate . The purpald assay provides a tool in addition to the existing ones for the measurement of glycol-containing PS . Among the usefulness of this method are the determinations of the glycerol content in the phospho-glycerol-containing PS and the SG and UG contents and structural integrity in PS and conjugate vaccines . Genetika, 2001 Jun, 37(6), 745 - 53 {Transgenic Tobacco plants expressing bacterial genes encoded the thermostable glucanases}; Movsesian NR et al.; It is shown that bacterial genes for thermostable beta-glucanases are expressed retaining their activity and substrate specificity . The leader peptide of the carrot extensin exerts effective secretion of the bacterial enzymes into the intercellular space of the plant tissue . Expression of the bacterial gene for beta-1,3-glucanase in plant tissues alters their morphogenetic potential . Regeneration of shoots from the calli of these plant lines requires a six- to eightfold increase in cytokinin (6-BAP) concentration in comparison with the control lines and the transgenic lines expressing beta-1,3-1,4-glucanase . Rooting of transgenic plants expressing the bacterial gene for beta-1,3-glucanase occurs much faster . The transgenic plants obtained in the study are proposed as model objects for investigating the role of glucanases in plants. Eur J Ultrasound, 2001 Jul, 13(3), 201 - 4 Hyperechoic focal bacterial nephritis: findings on contrast-enhanced Colour Doppler ultrasound; Esteban JM et al.; We report a case of atypical focal bacterial nephritis (lobar nephronia) simulating a renal mass on gray-scale ultrasound, describing the findings on Colour Doppler ultrasound before and after administration of a galactose-based ultrasound contrast agent. Infect Dis Obstet Gynecol, 2001, 9(3), 133 - 41 A multicenter study of bacterial vaginosis in women with or at risk for human immunodeficiency virus infection; Warren D et al.; BACKGROUND: Bacterial vaginosis is a common gynecologic infection that has been associated with a variety of gynecologic and obstetric complications, including pelvic inflammatory disease, postabortal infection and premature delivery . Recent studies suggest that bacterial vaginosis may increase a woman's risk for human immunodeficiency virus (HIV) . We undertook this study to assess whether the prevalence and characteristics of bacterial vaginosis differed according to HIV status in high-risk US women . METHODS: Prevalence of bacterial vaginosis was assessed by Gram's stain and clinical criteria for 854 HIV-infected and 434 HIV-uninfected women enrolled in the HIV Epidemiology Research (HER) Study . Multiple logistic regression techniques were used to determine whether HIV infection independently predicted bacterial vaginosis . RESULTS: Almost half (46%) the women had bacterial vaginosis by Gram's stain . The prevalence of bacterial vaginosis was 47% in the HIV-positive women compared with 44% in the HIV-negative women; this difference was not statistically significant (p = 0.36) . After adjustment for other covariates, HIV-positive women were more likely than HIV-negative women to have bacterial vaginosis (odds ratio (OR) 1.31; 95% confidence interval (CI) 1.01-1.70) by Gram's stain but not by clinical criteria (OR 1.16; CI 0.87-1.55) . Among HIV-positive women, use of antiretroviral drugs was associated with a lower prevalence of bacterial vaginosis (adjusted OR 0.54; Cl 0.38-0.77) . CONCLUSIONS: In this cross-sectional analysis of high-risk US women, HIV infection was positively correlated with bacterial vaginosis diagnosed by Gram's stain. Biotechniques, 2001 Aug, 31(2), 355, 358 - 62 Development of simple sequence repeat markers from bacterial artificial chromosomes without subcloning; Qi X et al.; Simple sequence repeats (SSRs) were isolated from pearl millet bacterial artificial clones (BACs) without any subcloning steps . SSR sequences were targeted using 3' end-anchored SSR primers . Flanking sequences were isolated by suppression PCR . In this pilot study, 25 SSR markers have been developed from 40 BAC pools, comprising a total of 384 clones . This novel way to develop new markers has the added advantage that mapping the SSR markers will anchor individual BACs to the genetic maps and, thus, facilitate the construction of BAC contigs. Genetics, 2001 Aug, 158(4), 1711 - 24 Bacterial artificial chromosome-based physical map of the rice genome constructed by restriction fingerprint analysis; Tao Q et al.; Genome-wide physical mapping with bacteria-based large-insert clones (e.g., BACs, PACs, and PBCs) promises to revolutionize genomics of large, complex genomes . To accelerate rice and other grass species genome research, we developed a genome-wide BAC-based map of the rice genome . The map consists of 298 BAC contigs and covers 419 Mb of the 430-Mb rice genome . Subsequent analysis indicated that the contigs constituting the map are accurate and reliable . Particularly important to proficiency were (1) a high-resolution, high-throughput DNA sequencing gel-based electrophoretic method for BAC fingerprinting, (2) the use of several complementary large-insert BAC libraries, and (3) computer-aided contig assembly . It has been demonstrated that the fingerprinting method is not significantly influenced by repeated sequences, genome size, and genome complexity . Use of several complementary libraries developed with different restriction enzymes minimized the "gaps" in the physical map . In contrast to previous estimates, a clonal coverage of 6.0-8.0 genome equivalents seems to be sufficient for development of a genome-wide physical map of approximately 95% genome coverage . This study indicates that genome-wide BAC-based physical maps can be developed quickly and economically for a variety of plant and animal species by restriction fingerprint analysis via DNA sequencing gel-based electrophoresis. Clin Infect Dis, 2001 Sep 15, 33(6), 894 - 6 Epub 2001 Aug 10. Association between