Arch AIDS Res, 1992, 6(3), 177 - 82 Bacterial vaginosis associated with G vaginallis / Mobiluncus sp: ultrastructural parameters; Villegas-castrejon H et al.; PIP: Physicians at the National Institute of Perinatology in Mexico City, Mexico used a Carl-Zeiss EM 10C electron microscope to examine genital secretion samples from 10 pregnant women (15-38 weeks' gestation) who had been diagnosed with Mobiluncus species and Gardnerella vaginalis infections to illustrate the form and structure of bacteria responsible for bacterial vaginosis . They were concerned that these bacteria induce preterm labor and premature rupture of membranes (PL/PROM) . These bacteria have been present in the genital tract of 30% of pregnant women with a thick whitish discharge who have attended the Institute's prenatal outpatient clinic . Physicians noted on the microscope slides that bacteria surrounded vaginal squamous epithelial cells (clue cells) . Numerous gardnerella-like bacteria surrounded elongated squamous epithelial cells with many plasma projections . An extensive area of lysis existed around the bacteria in the cytoplasm of many squamous epithelial cells with intact membrane and nonexistent microfilaments . This finding indicated that the bacteria invade and destroy the cells . Plasma membrane projections almost completely surrounded the gardnerella-like bacteria in certain areas . Since this study strengthened the theory that G . vaginalis enters the vaginal squamous epithelial cells, researchers should conduct more studies to determine its role in PL/PROM . Biofizika, 2002 Jul-Aug, 47(4), 595 - 9 {Similarities in periodical structures in the position of nucleotides in regions of initiation of replication of bacterial genomes}; Kravatskaia GI et al.; The regions of initiation of replication of some bacterial genomes were studied by the method of Fourier matrix analysis . A generalized spectral portrait of the primary structures of E . coli-like regions of initiation of replication in bacteria was obtained, which reflects the features of their structural and functional organization . It contains well-pronounced peaks that correspond to the periods T = 2, 11, 17, 27, 86-105 of nucleotides . The peaks corresponding to T = 9, 13, 14, 18, 19, 33-35, 45-47, 74-85, 106-110 are less pronounced . The uniqueness of the Fourier spectrum corresponding to the region of initiation of replication of E . coli oriC was considered by the example of the complete genome of E . coli . Some regions of the E . coli genome were identified that differ from oriC in the primary structure but have Fourier spectra resembling the spectrum of oriC . A number of these regions are alternative points of initiation of replication in sdrA(rnh) mutants of E . coli, the others are localized in yet unidentified regions of the E . coli genome but are capable, in our opinion, to participate in the initiation of replication . Thus, from the similarity of spectral portraits of different regions of the genome, it was possible to reveal several regions that have similar functions, i.e., are involved in initiation of replication. Biol Reprod, 2002 Oct, 67(4), 1337 - 41 Intrauterine bacterial inoculation induces labor in the mouse by mechanisms other than progesterone withdrawal; Hirsch E et al.; We tested the hypothesis that progesterone (P(4)) withdrawal is the primary mechanism by which intrauterine bacteria induce preterm labor in mice . CD-1 mice on Day 14.5 of a 19- to 20-day gestation were subjected to one of four treatments: 1) intrauterine injection of sterile medium, 2) intrauterine injection of 10(6) heat-killed Escherichia coli bacteria, 3) intrauterine injection of 10(9) heat-killed E . coli, or 4) ovariectomy . Mice were then killed at four time points from 0.75 to 11 h after surgery for serum collection . Separately, animals were pretreated either with s.c . P(4) or with vehicle 2 h before ovariectomy or high-dose bacterial inoculation . Ovariectomy led to a rapid fall in serum P(4) levels of 60% by 1 h and 81% by 8 h compared with levels in controls (P < 0.001) . In contrast, intrauterine inoculation with 10(9) bacteria led to a more modest decline in P(4) of only 28% by 8 h (P = 0.24, which was no different from that of 10(6) bacteria, an inoculum below the threshold for preterm delivery) . Despite significantly lower levels of P(4) in the ovariectomy group, time to delivery was significantly shorter with 10(9) bacteria intrauterine (24 +/- 5.6 h vs . 19 +/- 3.6 h, P = 0.03) . Pretreatment with 1.5 mg P(4) per mouse prolonged the interval to delivery following both ovariectomy and high-dose bacteria, in association with pharmacologically elevated serum P(4) levels . In contrast, physiologic P(4) supplementation (0.375 mg/mouse) prolonged gestation only in the ovariectomy group . We conclude that withdrawal of endogenous P(4) is not the primary cause of labor following intrauterine bacterial inoculation in mice. Circulation, 2002 Sep 24, 106(13), 1659 - 63 Serum immunoglobulin G antibodies to chlamydial heat shock protein 60 but not to human and bacterial homologs are associated with coronary artery disease; Mahdi OS et al.; BACKGROUND: Evidence for an association between Chlamydia pneumoniae infection and coronary artery disease (CAD) has been reported by numerous studies, cross-reactive heat shock protein (Hsp) antibody responses have been causally linked to CAD, and the severity of chlamydial disease pathogenesis correlates with Hsp serology . Our aim was to determine if chlamydial Hsp (cHsp) antibody responses are predictive of CAD . METHODS AND RESULTS: Patients were recruited in a case-control study: 250 cases had angiographically significant CAD (stenosis > or =70%), and 250 controls had normal coronary arteries (stenosis <10%) . Serum immunoglobulin G reactivity to Hsp10 and Hsp60 antigens (chlamydial, Escherichia coli, and human), and C pneumoniae whole organisms were measured by ELISA . Univariate analysis confirmed that classical CAD risk factors were predictors of CAD . Univariate analysis showed that cHsp60 (P= 0.001, OR 3.9), cHsp10 (P=0.045, OR 3.8), E coli Hsp60 (P=0.04, OR 1.5) and C pneumoniae (P=0.03, OR 1.8) ELISA optical density (OD) values were significantly different between cases and controls . Multivariate analysis found that only upper-quintile cHsp60 seroreactivity remained a significant predictor of CAD after controlling for classical CAD risk factors and seroreactivity to the other antigens (cHsp60 OD, P=0.005, OR 3.9 per OD unit; cHsp60 quintile, 5 versus 1 to 4; P=0.01, OR 2.1) . CONCLUSIONS: The presence of elevated anti-cHsp60 immunoglobulin G antibodies, but not anti-human or anti-E coli homologs, was independently associated with CAD . This finding argues against previous suggestions that cross-reactive or autoimmune Hsp60 responses may contribute to disease progression . High anti-cHsp60 antibody response appears to identify the subset of patients with chlamydial infection and significant CAD. Neurosci Lett, 2002 Sep 27, 330(3), 270 - 4 The bacterial chaperonin GroEL requires GroES to reduce aggregation and cell death in a COS-7 cell model of Huntington's disease; Carmichael J et al.; Huntington's disease (HD) is caused by expansions of more than 35 CAG repeats in the HD gene . These repeats are translated into a long polyglutamine tract that confers a deleterious gain-of-function on the mutant protein . Intraneuronal inclusions comprising mutant huntingtin are found in HD patient brains . Here we show that the bacterial chaperonin GroEL can reduce aggregation of mutant huntingtin in COS-7 cells and requires GroES for efficient activity, analogous to what has been described in bacteria . The reduction in aggregation of mutant huntingtin by GroEL/GroES was associated with protection against polyglutamine-induced cell death . Eur J Obstet Gynecol Reprod Biol, 2002 Oct 10, 105(1), 64 - 6 Local treatment of bacterial vaginosis with a bioadhesive metronidazole tablet; Voorspoels J et al.; OBJECTIVES: Oral metronidazole is still the drug of choice in the treatment of bacterial vaginosis . Yet, side effects have been reported, and dosage as well as duration of therapy are still controversial . This study presents a possible alternative treatment using a single dose of metronidazole administered in a vaginal bioadhesive tablet . STUDY DESIGN: Double blind, randomised, placebo-controlled trial . SUBJECTS: The 116 patients were allocated to placebo; metronidazole 100; 250; 500mg in a 1:1:1:1 ratio . RESULTS: A cure rate of 64% was obtained with a single 100mg dose of metronidazole formulated in a bioadhesive vaginal tablet compared to a cure rate of 29% in the placebo group . The cure rates with the higher doses were similar 61.5% for 250mg dose and 68% for the 500mg dose . No side effects were reported . CONCLUSIONS: Treatment of bacterial vaginosis with a single application of 100mg metronidazole in a bioadhesive vaginal tablet was found to be a valid alternative . Further research in relation to tablet shaping and optimal dose finding might increase the cure rate. Mikrobiologiia, 2002 Jul-Aug, 71(4), 437 - 44 {Aspects of cellular biology of the bacterial stationary phase: programmed cell death and regulation by guanosine tetraphosphate}; Golovlev EL; The paper discusses (1) programmed cell death, the phenomenon typical of the stationary phase of bacteria occurring under unfavorable conditions, (2) its pleiotropic regulation by guanosine tetraphosphate, and (3) the conception of "addiction module," a specific genetic system responsible for the cell choice between survival and death under unfavorable conditions . The shortcomings of the proposed interpretation of the problem at hand are considered and the necessity of their further investigation is substantiated. J Immunol, 2002 Oct 1, 169(7), 3934 - 9 IL-1 receptor-associated kinase and low molecular weight GTPase RhoA signal molecules are required for bacterial lipopolysaccharide-induced cytokine gene transcription; Chen LY et al.; Proinflammatory cytokines such as IL-1, TNF, IL-6, and IL-8 are produced by leukocytes in response to bacteria or bacterial components . A great deal has been learned during the past few years about the synthesis and release of proinflammatory cytokines by leukocytes; however, relatively little is known about the intracellular events that lead to leukocyte proinflammatory cytokine gene transcription . This study examined the signal transduction pathway of IL-8 induction by bacterial LPS . Stimulation of monocytes with LPS rapidly activated RhoA, and pretreatment of monocytes with a RhoA inhibitor, C3 transferase exoenzyme, effectively blocked LPS-induced IL-8 gene expression . Overexpression of dominant negative RhoA (T19N) or IL-1R-associated kinase completely inhibited LPS-stimulated reporter gene expression . Induction of IL-8 was also inhibited by dominant negative IkappaB kinase and myeloid differentiation protein (MyD88) . These results indicate that RhoA and IL-1R-associated kinase are novel signal transducers for LPS-induced Toll-like receptor 4-mediated proinflammatory cytokine synthesis in human monocytes. Avian Dis, 2002 Jul-Sep, 46(3), 605 - 12 Studies on the bacterial etiology of airsacculitis of broilers in northern and middle Jordan with special reference to Escherichia coli, Ornithobacterium rhinotracheale, and Bordetella avium; El-Sukhon SN et al.; A total of 100 poultry farms in northern and middle areas of Jordan were sampled to investigate the bacteria associated with airsacculitis in broiler chickens . Of 170 bacterial isolates, 88.2% were identified as Escherichia coli, 8.8% as Ornithobacterium rhinotracheale, and 3% as Bordetella avium . Fourteen serotypes of E . coli were identified among 66 typeable isolates and the remainder were untypeable . The most prevalent serotypes were O1, O8, and O78 . The main serotype of O . rhinotracheale was serotype A . Experimental inoculation of O . rhinotracheale via intravenous, intratracheal, and intra-air sac routes resulted in growth retardation, thickening in the air sacs, arthritis, and liver necrosis . Reisolation of O . rhinotracheale from the air sacs, liver, trachea, heart, and spleen at day 7 postinoculation confirmed its role . In vitro susceptibility testing revealed that E . coli isolates were sensitive to gentamicin and colistin, O . rhinotracheale to tetracyline, and B . avium to most of the nine antibiotics examined. Plant Cell, 1995 Jan, 7(1), 29 - 42 Coordinated Activation of Programmed Cell Death and Defense Mechanisms in Transgenic Tobacco Plants Expressing a Bacterial Proton Pump; Mittler R et al.; In plants, programmed cell death is thought to be activated during the hypersensitive response to certain avirulent pathogens and in the course of several differentiation processes . We describe a transgenic model system that mimics the activation of programmed cell death in higher plants . In this system, expression of a bacterial proton pump in transgenic tobacco plants activates a cell death pathway that may be similar to that triggered by recognition of an incompatible pathogen . Thus, spontaneous lesions that resemble hypersensitive response lesions are formed, multiple defense mechanisms are apparently activated, and systemic resistance is induced in the absence of a pathogen . Interestingly, mutation of a single amino acid in the putative channel of this proton pump renders it inactive with respect to lesion formation and induction of resistance to pathogen challenge . This transgenic model system may provide insights into the mechanisms involved in mediating cell death in higher plants . In addition, it may also be used as a general agronomic tool to enhance disease protection. Genetics, 2002 Sep, 162(1), 129 - 34 A bacterial artificial chromosome-based genetic linkage map of the nematode Pristionchus pacificus; Srinivasan J et al.; To understand the evolution of developmental processes, nonmodel organisms in the nematodes, insects, and vertebrates are compared with established model systems . Often, these comparisons suffer from the inability to apply sophisticated technologies to these nonmodel species . In the nematode Pristionchus pacificus, cellular and genetic analyses are used to compare vulva development to that of Caenorhabditis elegans . However, substantial changes in gene function between P . pacificus and C . elegans limit the use of candidate gene approaches in studying P . pacificus mutations . To facilitate map-based cloning of mutations in P . pacificus, we constructed a BAC-based genetic linkage map . A BAC library of 13,440 clones was generated and completely end sequenced . By comparing BAC end and EST sequences between the "wild-type" strain P . pacificus var . California and the polymorphic strain P . pacificus var . Washington, 133 single-stranded conformational polymorphisms were identified . These markers were tested on a meiotic mapping panel of 46 randomly picked F(2) animals after a cross of the two strains, providing the first genetic linkage map of P . pacificus . A mapping strategy using two selected markers per chromosome was devised and the efficiency of this approach was illustrated by the mapping of the Ppa-unc-1/Twitchin gene. Biotechnol Appl Biochem, 2002 Oct, 36(Pt 2), 85 - 8 Guide DNA technique in bacterial ribonuclease P reaction for effective processing of tRNA precursor; Tanaka T et al.; Previously, we found that a small (approx . 20-mer) DNA hybridizing to the 5'-leader region of a tRNA precursor enhances the cleavage efficiency in bacterial ribonuclease P reaction . We named this technique the 'guide DNA technique' . Detailed analyses showed that the length of the guide DNA, concentration of the guide DNA and the hybridizing position affected the cleavage efficiency: for an effective cleavage reaction, guide DNA should be designed to hybridize to the region on the cleavage site, should be 20 bases or more in length and should be of high concentration . The presence of a 5'-flanking region in the DNA did not affect the cleavage reaction . The guide DNA technique is a useful tool for effective preparation of mature tRNA molecules in vitro. Mol Biol Rep, 2002, 29(1-2), 21 - 6 Flux control of the bacterial phosphoenolpyruvate:glucose phosphotransferase system and the effect of diffusion; Francke C et al.; We analyzed the role of diffusion and cell size on the flux control properties of the glucose-PTS of Escherichia coli, in silicon cells under various metabolic conditions . To our surprise, the influence of the concentration of phosphoryl-donor PEP on the distribution of control was small . We found for cells of bacterial size that PTS-flux control was mainly located in processes taking place in the membrane and that diffusion hardly controlled the flux (< 2.8 %) . Enlargement of the cells shifted the control from membrane to cytoplasm and from process rates to diffusion rates, the latter now having a total control of about 38 % . In the presence of glucose, nearly all diffusion flux control resided in the component that links the cytoplasmic processes to those in the membrane. Protein Sci, 2002 Oct, 11(10), 2512 - 21 Bacterial expression and membrane targeting of the rat complement regulator Crry: a new model anticomplement therapeutic; Fraser DA et al.; Inappropriate or unregulated activation of complement can contribute to pathology in inflammatory diseases . Previous studies have shown that soluble recombinant regulators of complement are effective in animal models and some human diseases . However, limitations include cost, rapid clearance, and unwanted systemic effects . To avoid some of these problems, bacterial expression of regulators has been optimized and methods for the addition of a membrane-targeting moiety to the complement regulator developed . When administered directly to sites of inflammation, membrane-targeted human regulators are retained and inhibit complement-activation locally . To test the efficacy of membrane-targeted complement regulators in vivo, we have undertaken the expression and membrane targeting of the rat-complement regulator Crry . A soluble recombinant form of Crry, containing only the first four short consensus repeats, was expressed in a mammalian expression system and shown to be functional as a fluid phase regulator . To generate the quantities required for testing in vivo, Crry was expressed in bacteria and refolded successfully . Refolded protein had full-complement regulatory activity in vitro . Attachment of a membrane address tag conferred membrane-binding capacity and greatly increased complement regulatory function in vitro . This novel anticomplement agent can now be applied to rat models of arthritis and other inflammatory diseases. Rev Esp Cardiol, 2002 Sep, 55(9), 988 - 90 {Tricuspid stenosis after pacemaker implantation without evidence of bacterial endocarditis . A case report}; Garrote C et al.; Tricuspid stenosis related to endocardial pacemaker leads is uncommon . We report the case of a patient with severe tricuspid stenosis documented 15 years after the implantation of a permanent DDD pacemaker for symptomatic congenital heart block . The atrial and ventricular leads both had a loop at the level of the tricuspid valve that may have caused endothelial damage and, eventually, tricuspid stenosis. Anal Chem, 2002 Sep 1, 74(17), 4410 - 6 On-probe digestion of bacterial proteins for MALDI-MS; Harris WA et al.; On-probe digestion combined with MALDI mass spectrometry is studied as a rapid method for the analysis and identification of bacterial proteins . The use of trypsin adsorbed to the probe surface reduces the digestion time from hours to minutes . A high amount of trypsin must be applied to the probe for the successful digestion of bacterial proteins . Mass spectra of the digest contain a number of low-mass digest fragments . Several components of a B . subtilis bacterial digest can be identified through postsource decay and database searching. EMBO J, 2002 Sep 16, 21(18), 4763 - 73 The structure of bacterial DnaA: implications for general mechanisms underlying DNA replication initiation; Erzberger JP et al.; The initiation of DNA replication is a key event in the cell cycle of all organisms . In bacteria, replication initiation occurs at specific origin sequences that are recognized and processed by an oligomeric complex of the initiator protein DnaA . We have determined the structure of the conserved core of the Aquifex aeolicus DnaA protein to 2.7 A resolution . The protein comprises an AAA+ nucleotide-binding fold linked through a long, helical connector to an all-helical DNA-binding domain . The structure serves as a template for understanding the physical consequences of a variety of DnaA mutations, and conserved motifs in the protein suggest how two critical aspects of origin processing, DNA binding and homo-oligomerization, are mediated . The spatial arrangement of these motifs in DnaA is similar to that of the eukaryotic-like archaeal replication initiation factor Cdc6/Orc1, demonstrating that mechanistic elements of origin processing may be conserved across bacterial, archaeal and eukaryotic domains of life. J Appl Microbiol, 2002, 93(4), 656 - 67 An arsenic(III)-oxidizing bacterial population: selection, characterization, and performance in reactors; Battaglia-Brunet F et al.; AIMS: To select an autotrophic arsenic(III)-oxidizing population, named CASO1, and to evaluate the performance of the selected bacteria in reactors . METHODS AND RESULTS: An As(III)-containing medium without organic substrate was used to select CASO1 from a mining environment . As(III) oxidation was studied under batch and continuous conditions . The main organisms present in CASO1 were identified with molecular biology tools . CASO1 exhibited significant As(III)-oxidizing activity between pH 3 and 8 . The optimum temperature was 25 degrees C . As(III) oxidation was still observed in the presence of 1000 mg l(-1) As(III) . In continuous culture mode, the As(III) oxidation rate reached 160 mg l(-1) h(-1) . The CASO1 consortium contains at least two organisms - strain b3, which is phylogenetically close to Ralstonia picketii, and strain b6, which is related to the genus Thiomonas . The divergence in 16S rDNA sequences between b6 and the closest related organism was 5.9%, suggesting that b6 may be a new species . CONCLUSIONS: High As(III)-oxidizing activity can be obtained without organic nutrient supply, using a bacterial population from a mining environment . SIGNIFICANCE AND IMPACT OF THE STUDY: The biological oxidation of arsenite by the CASO1 population is of particular interest for decontamination of arsenic-contaminated waste or groundwater. Biochem Cell Biol, 2002, 80(4), 415 - 20 Screening regulatory sequences from bacterial artificial chromosome DNA of alpha- and beta-globin gene clusters; Zhang S et al.; In the forthcoming postgenomic era, identification of regulatory DNA sequences is becoming increasingly important for characterizing DNA-binding proteins and for elucidating the regulatory mechanisms of gene expression . Presently, there lack efficient methods to broadly screen and identify DNA regulatory elements on a large scale . We established herein an efficient strategy to screen regulatory sequences from bacterial artificial chromosome (BAC) DNAs containing human alpha- and beta-globin gene clusters based on polymerase chain reaction and electrophoretic mobility shift assay (EMSA) techniques without purified transcription factors . Twenty-three subclones derived from alpha-BAC DNA by bulk EMSA selection retained the ability to bind nuclear proteins of K562 cells when retested by EMSA . In 19 clones sequenced, 14 are identical to those registered in GenBank and five have one base difference . All of the 24 randomly picked beta-BAC clones showed specific binding with nuclear proteins of K562 cells . In 11 clones sequenced, eight are identical to those registered in GenBank and three have one base difference . This approach could be particularly powerful if combined with other systematic methods for identifying cis-regulatory DNA elements. Plant Physiol, 1994 Jun, 105(2), 473 - 482 Plant Expression of a Bacterial Cytochrome P450 That Catalyzes Activation of a Sulfonylurea Pro-Herbicide; O'Keefe DP et al.; The Streptomyces griseolus gene encoding herbicide-metabolizing cytochrome P450SU1 (CYP105A1) was expressed in transgenic tobacco (Nicotiana tabacum) . Because this P450 can be reduced by plant chloroplast ferredoxin in vitro, chloroplast-targeted and nontargeted expression were compared . Whereas P450SU1 antigen was found in the transgenic plants regardless of the targeting, only those with chloroplast-directed enzyme performed P450SU1-mediated N-dealkylation of the sulfonylurea 2-methylethyl-2,3-dihydro-N-{(4,6-dimethoxypyrimidin-2-yl)aminocarbonyl}-1, 2-benzoisothiazole- 7-sulfonamide-1,1-dioxide (R7402) . Chloroplast targeting appears to be essential for the bacterial P450 to function in the plant . Because the R7402 metabolite has greater phytotoxicity than R7402 itself, plants bearing active P450SU1 are susceptible to injury from R7402 treatment that is harmless to plants without P450SU1 . Thus, P450SU1 expression and R7402 treatment can be used as a negative selection system in plants . Furthermore, expression of P450SU1 from a tissue-specific promoter can sequester production of the phytotoxic R7402 metabolite to a single plant tissue . In tobacco expressing P450SU1 from a tapetum-specific promoter, treatment of immature flower buds with R7402 caused dramatically lowered pollen viability . Such treatment could be the basis for a chemical hybridizing agent. Plant Physiol, 1994 Jan, 104(1), 153 - 159 Aminomethylenediphosphonate: A Potent Type-Specific Inhibitor of Both Plant and Phototrophic Bacterial H+-Pyrophosphatases; Zhen RG et al.; The suitability of different pyrophosphate (PPi) analogs as inhibitors of the vacuolar H+-translocating inorganic pyrophosphatase (V-PPase; EC 3.6.1.1) of tonoplast vesicles isolated from etiolated hypocotyls of Vigna radiata was investigated . Five 1,1-diphosphonates and imidodiphosphate were tested for their effects on substrate hydrolysis by the V-PPase at a substrate concentration corresponding to the Km of the enzyme . The order of inhibitory potency (apparent inhibition constants, Kiapp values, {mu}M, in parentheses) of the compounds examined was aminomethylenediphosphonate (1.8) > hydroxymethylenediphosphonate (5.7) {almost equal to} ethane-1-hydroxy-1,1-diphosphonate (6.5) > imidodiphosphate (12) > methylenediphosphonate (68) >> dichloromethylenediphosphonate (>500) . The specificity of three of these compounds, aminomethylenediphosphonate, imidodiphosphate, and methylenediphosphonate, was determined by comparing their effects on the V-PPase and vacuolar H+-ATPase from Vigna, plasma membrane H+-ATPase from Beta vulgaris, H+-PPi synthase of chromatophores prepared from Rhodospirillum rubrum, soluble PPase from Saccharomyces cerevisiae, alkaline phosphatase from bovine intestinal mucosa, and nonspecific monophosphoesterase from Vigna at a PPi concentration equivalent to 10 times the Km of the V-PPase . Although all three PPi analogs inhibited the plant V-PPase and bacterial H+-PPi synthase with qualitatively similar kinetics, whether substrate hydrolysis or PPi-dependent H+-translocation was measured, neither the vacuolar H+-ATPase nor plasma membrane H+-ATPase nor any of the non-V-PPase-related PPi hydrolases were markedly inhibited under these conditions . It is concluded that 1, 1-diphosphonates, in general, and aminomethylenediphosphonate, in particular, are potent type-specific inhibitors of the V-PPase and its putative bacterial homolog, the H+-PPi synthase of Rhodospirillum. Trop Med Int Health, 2002 Sep, 7(9), 722 - 31 Clinical predictors of bacterial meningitis in infants and young children in The Gambia; Weber MW et al.; BACKGROUND: Bacterial meningitis is an important cause of childhood morbidity and mortality world-wide . In the developing world, where the burden of acute meningitis and its long-term sequelae are especially high, staff with limited training at primary health care facilities must be able to recognize the symptoms and signs of meningitis, so that suspected cases can be referred urgently to hospitals . METHODS: Children who presented with possible invasive bacterial infection to health facilities in The Gambia, West Africa, between 1993 and 1995 were investigated in a standardized manner and clinical findings were documented . Bacterial meningitis was defined as the growth of bacteria from the cerebrospinal fluid . Clinical findings were compared between cases of meningitis and other children . RESULTS: Of 2097 children between 2 months and 3 years of age investigated, 51 had a confirmed diagnosis of bacterial meningitis . In multivariate analysis using a model adjusting for age but not including respiratory signs, the variables associated independently with meningitis were appearance of being very sick (odds ratio for meningitis vs . no meningitis or no lumbar puncture performed (OR) 4.1, 95% CI 1.5-11.1), being lethargic or unconscious (OR 5.2, 95% CI 2.1-13), a stiff neck (OR 29.3, 95% CI 12.2-70.3), a bulging fontanel (OR 3.2, 95% CI 1.2-8.5) and reduced feeding as a prompted complaint (OR 2.9, 95% CI 1.3-6.7) . A combination model of a history of convulsions, or being lethargic or unconscious, or having a stiff neck, as used in the WHO-Integrated Management of Childhood Illness (IMCI) guidelines, had a sensitivity of 98% and a specificity of 72% to predict meningitis . CONCLUSIONS: A combination of a limited number of signs is sufficient to predict meningitis with high sensitivity, without a large number of children who do not have meningitis being unnecessarily referred. Phys Rev Lett . 2002 Sep 9;89(11):118102 . Epub 2002 Aug 23. Periodic chirality transformations propagating on bacterial flagella; Coombs D et al.; When a helical bacterial flagellum, clamped at one end, is placed in an external flow, it has been observed that regions of the flagellum transform to the opposite chirality, and travel as pulses down the length of the filament, the process repeating periodically {H . Hotani, J . Mol . Biol . 156, 791 (1982)}} . We propose a theory for this phenomenon based on a treatment of the flagellum as an elastic object with multiple stable configurations . The simplest possible implementation of the model accurately reproduces key features seen in experiment. Biol Chem, 2002 Jun, 383(6), 945 - 60 The bacterial regulatory protein H-NS--a versatile modulator of nucleic acid structures; Schroder O et al.; The small DNA binding protein H-NS is attracting broad interest for its profound involvement in the regulation of bacterial physiology . It is involved in the regulation of many genes in response to a changing environment and functions in the adaptation to many different kinds of stress . Many H-NS-controlled genes, including the hns gene itself, are further linked to global regulatory networks . H-NS thus plays a key role in maintaining bacterial homeostasis under conditions of a rapidly changing environment . In this review we summarize recent results from combined biochemical and biophysical efforts which have yielded new insights into the three-dimensional structure and function of H-NS . The protein consists of two distinct domains separated by an unstructured linker region, and the structural details available today have helped to understand how these domains may interact with each other or with ligand molecules . Functional studies have, in addition, revealed mechanistic clues for the various H-NS activities, like temperature- or growth phase-dependent regulation . Important elements for the specific regulatory activities of H-NS comprise different modes of DNA binding, protein oligomerization, the competition with other regulators and the fact that the topology of the target DNA is modulated during complex formation . The distinctive ability to recognize nucleic acid structures in combination with other proteins also explains H-NS-dependent post-transcriptional activities where the interaction with defined RNA structures and the interference with RNA/protein complexes during mRNA translation are crucial for regulation . Thus, protein/protein interactions, in combination with the recognition and modulation of nucleic acid structures, are key elements of the different mechanisms which make H-NS such a versatile regulator. Med Sci Monit, 2002 Sep, 8(9), BR345 - 53 Repeated intravenous doses of all-trans-retinoyl beta-D-glucuronide is not effective in the treatment of bacterial bronchopneumonia in lambs but is devoid of gross and acute toxicity; Gallup JM et al.; BACKGROUND: All-trans-retinoyl beta-D-glucuronide, a water-soluble glucuronic acid conjugate of all-trans-retinoic acid, has demonstrated high biological activity and low toxicity in most in vitro and in vivo models . Since the reparative effects of retinoids on epithelium are well-known, our aim was to study the effect(s) of intravenously-administered all-trans-retinoyl beta-D-glucuronide in lambs with chronic bacterial bronchopneumonia . MATERIAL/METHODS: Two groups of lambs were inoculated intrabronchially with either pyrogen-free saline or Mannheimia haemolytica . Thirty-three days later, lambs were injected four times at five-day intervals with 2 mL of 116 mM all-trans-retinoyl beta-D-glucuronide (6.0-9.3 mmol/kg or 2.86-4.42 mg/kg animal body weight) in dimethyl sulfoxide, or dimethyl sulfoxide alone . Animal behavior and signs of clinical illness were logged daily . Lung and liver samples were assessed for histopathology, while serum and liver samples were extracted for retinoids and analyzed by reversed-phase gradient high-performance liquid chromatography . RESULTS: Repeated injections of highly concentrated all-trans-retinoyl beta-D-glucuronide did not cause changes in appetite, activity or other behaviors nor did it cause histologic lesions in liver and lung . However, it had no effect on resolution of lung lesions resultant of chronic Mannheimia haemolytica bronchopneumonia . CONCLUSIONS: Repeated intravenous administration of high amounts of all-trans-retinoyl beta-D-glucuronide to lambs did not elicit signs of gross or microscopic toxicity . However, administering all-trans-retinoyl beta-D-glucuronide too late in the progression of bacterial pneumonia is thought to be the main reason for its lack of effect in this model . A retinoid lactone derivative was detected in sheep serum and liver several days after treatment. J Hepatol, 2002 Oct, 37(4), 463 - 70 Bacterial infection in cirrhosis impairs coagulation by a heparin effect: a prospective study; Montalto P et al.; BACKGROUND: Bacterial infections have been postulated as a trigger for variceal bleeding in cirrhotic patients, and impair coagulation evaluated by thrombelastography (TEG) . Endogenous heparinoids have been detected after variceal bleeding and during liver transplantation in some cirrhotics using heparinase-modified-TEG . AIM: To assess if bacterial infection is associated with endogenous heparinoids in cirrhotics, thus impairing coagulation . METHODS: Native and heparinase-modified-TEG (cleavage of heparin and heparan-sulphate) was performed in 60 cirrhotics (Grade A, 2; B, 30; C, 28): 30 infected {septicaemia, 6 (culture positive); 6 (culture negative); spontaneous bacterial peritonitis, 10; chest infection, 4; others, 4}, 30 not infected, and five infected patients without liver diseases, comparing TEG parameters r, alpha, and ma . Eight cirrhotics were studied before and after infection . The diagnosis of presence and type of infection was based on international standard criteria . RESULTS: A significant heparin effect was found only in infected cirrhotics (28 of 30) with significant changes in r (P=0.0003), alpha (P<0.0001), and ma (P<0.0001), but in none of those not infected . This effect completely reversed in the eight evaluated after resolution of infection . There was no heparin effect in infected non-cirrhotics . CONCLUSIONS: A heparin effect was only found in cirrhotic patients with infection, further confirming that infection significantly modifies coagulation in cirrhotic patients. Biomacromolecules, 2002 Sep-Oct, 3(5), 1071 - 7 Studies on comonomer compositional distribution of bacterial poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)s and thermal characteristics of their factions; Feng L et al.; The comonomer-unit compositional distributions have been investigated for bacterial poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) {P(3HB-co-3HH)} samples with 3HH unit content of 13.8, 18.0, 22.0, and 54.0 mol % . They were comonomer compositionally fractionated using chloroform/n-heptane mixed solvent at ambient temperature . The fractionation of P(3HB-co-18.0 mol %3HH) and P(3HB-co-22.0 mol % 3HH), which could not be carried out effectively at room temperature, were refractionated at 70 degrees C in the mixed solvent . Fractions with different 3HH unit content in a wide range (from 4.4 to 80.7 mol %) were obtained . By use of these fractions with narrow compositional distribution, the comonomer composition dependence of thermal properties was investigated by differential scanning calorimetry . The melting point (T(m)) and heat of fusion (DeltaH) decreased as the 3HH unit content increased in the range of low 3HH content (<40 mol %), while they increased as the 3HH unit content increased in the high 3HH content range (>70 mol %) . The minimum T(m) and DeltaH values were found to exist at 3HH unit content of about 60 mol % . The glass transition temperature (T(g)) decreased linearly with the increase of 3HH unit content . The values of T(m), DeltaH, and T(g) of P(3HB-co-3HH)s were compared with those of poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3-hydroxybutyrate-co-3-hydroxypropionate), and poly(3-hydroxybutyrate-co-4-hydroxybutyrate), and the effects of comonomer types on the thermal properties were revealed. Biomacromolecules, 2002 Sep-Oct, 3(5), 1057 - 64 One-step synthesis of amphiphilic diblock copolymers from bacterial poly({R}-3-hydroxybutyric acid); Ravenelle F et al.; Catalyzed transesterification in the melt is used to produce diblock copolymers of poly({R}-3-hydroxybutyric acid), PHB, and monomethoxy poly(ethylene glycol), mPEG, in a one-step process . Bacterial PHB of high molecular weight is depolymerized by consecutive and partly simultaneous reactions: pyrolysis and transesterification . The formation of diblocks is accomplished by the nucleophilic attack from the hydroxyl end-group of the mPEG catalyzed by bis(2-ethylhexanoate) tin . The resulting diblock copolymers are amphiphilic and self-assemble into sterically stabilized colloidal suspensions of PHB crystalline lamellae. Eur J Gynaecol Oncol, 2002, 23(4), 366 - 8 Absence of bacterial growth in the culture from the epidural catheter of a patient with endometrial carcinoma and febrile neutropenia: a case report and review of the literature; Pirbudak L et al.; Infection is a potentially serious complication of long-term epidural (EP) catheterization in cancer patients . Although the use of epidural opioid analgesia is an effective and safe means for pain relief in terminally ill patients, these patients are in need of monitorization for possible infection . This is the first report in which EP catheter cultivation has been assessed in an immunocompromised and febrile neutropenic endometrial cancer patient. Acta Neurochir (Wien), 2002 Jun, 144(6), 601 - 8; discussion 608-9 Cerebral physiological and biochemical changes during vasogenic brain oedema induced by intrathecal injection of bacterial lipopolysaccharides in piglets; Gardenfors A et al.; BACKGROUND: The objective of the study was to evaluate biochemical and physiological changes in an experimental model of vasogenic brain oedema utilising techniques also used in routine neurointensive care . METHOD: 32 piglets were randomised to control or experimental group . The latter received an intrathecal injection of lipopolysaccharide (LPS) from E . coli (LPS group) . Intracranial pressure (ICP)and mean arterial pressure (MAP) were measured continuously . Intracerebral microdialysis was used for analysing interstitial levels of glucose, pyruvate, lactate, glutamate, glycerol and urea every 30 min . Repeated calculations of mean hemispheric CBF were performed utilising an extracranial scintillation detector and Intra-carotid injection of (133)Xe . Cerebral specific gravity was measured and the brains were fixed for histological examinations.FINDINGS: After LPS injection ICP increased reaching a plateau phase after 4-7 hours and CBF increased by 46% . Histological examination showed inflammation with pronounced extravasation of granulocytes . A significant decrease in brain specific gravity (p =0.022) was obtained . LPS caused a significant decrease in cerebral interstitial concentration of glucose (p = 0.0035), and significant increases in lactate concentration (p = 0.002) and lactate/pyruvate ratio (p = 0.0017) . A small but significant increase in glutamate was obtained (p = 0.0219) . Glycerol did not change significantly . INTERPRETATION: Intrathecal LPS caused an inflammatory reaction with extravasation of granulocytes, increased blood-brain barrier permeability and cerebral oedema . Biochemical analyses indicate increased glycolysis but no signs of cell membrane degradation. Med Sci Monit, 2001 May, 7 Suppl 1, 169 - 74 Influence of HBV or HCV associated chronic liver diseases on the course and outcome of purulent, bacterial meningoencephalitis; Kepa L et al.; BACKGROUND: Aim of the study was evaluation of HBV or HCV associated chronic liver diseases (HBV or HCV CHLD) influence on the course and outcome of purulent, bacterial meningoencephalitis (PBME), without symptoms of sepsis . MATERIAL AND METHODS: Between 1995-99 there were 8 patients with PBME, with chronic HBV (5 subjects) or HCV (3 subjects) infection, treated in our centre; mean age 43 years . Str . pneumoniae and N . meningitidis were etiologic factors of PBME in 25% and 12.5% of patients, respectively . In 62.5% of cases etiology of PBME remained unknown . In 2 patients HBV or HCV CHLD was diagnosed before PBME (1 case--chronic active hepatitis, 1 case--postinflammatory liver cirrhosis) . During hospitalization due to PBME in 4 patients liver cirrhosis was diagnosed on the base of clinical picture and laboratory results, in 2 patients chronic hepatitis B or C was subject to further diagnostic procedures . RESULTS: In 7 subjects (87.5%) significant increase of AlAT and AspAT activity was recorded during acute phase of neuroinfection as compared to results preceding the hospitalization (to 300-400 U/l) . Together with recovery from PBME decrease of aminotransferases activity was noted . In 1 fatal case high AlAT and AspAT activity was observed for the whole time of the disease . In 2 other patients with liver cirrhosis, classified into class A of Child-Turcott-Pugh classification at the beginning of PBME, after transient insignificant aminotransferases increase sudden decompensation of liver functions during recovery from PBME was observed . Both patients died due to haemorrhage from esophageal varices . In all patients with PBME and HBV or HCV CHLD inflammatory parameters of cerebrospinal fluid were increased for longer than average time . It was the reason of longer hospital stay . The influence of HBV or HCV CHLD on PBME outcome was not observed . CONCLUSIONS: 1 . In patients with PBME concomitant HBV or HCV CHLD may exert negative influence on the course of neuroinfection and extend the period of hospitalization . 2 . The increase of aminotransferases activity in these patients may suggest other hepatotropic virus superinfection and require further diagnostics . 3 . In the case of HBV or HCV associated postinflammatory liver cirrhosis PBME may be connected with rapid liver disease progression and even the death of a patient. Chembiochem, 2002 Sep 2, 3(9), 874 - 86 Aspartyl phosphonates and phosphoramidates: the first synthetic inhibitors of bacterial aspartate-semialdehyde dehydrogenase; Cox RJ et al.; The synthesis of methylene phosphonate, difluoromethylene phosphonate and phosphoramidate analogues of aspartyl phosphate, together with reduced analogues, is described . These compounds were shown to be effective inhibitors of aspartate-semialdehyde dehydrogenase (ASA-DH) from Escherichia coli . However, despite the structural similarity of the compounds, different patterns of inhibition were observed, indicative of two phases of recognition and binding . Correlation between measured inhibition constants with pK(a) values supports the theory that binding at the phosphate binding site is optimised for singly ionised phosphate analogues. Pediatr Pulmonol, 2002 Oct, 34(4), 324 - 8 Recurrent unilateral bacterial pneumonias and interstitial fibrosis associated with pulmonary vein atresia: successful treatment with endovascular stent implantation; Sacco O et al.; A variety of pulmonary vascular disorders, such as hemangiomatosis, telangectasia, and veno-occlusive disease, may be involved in the pathogenesis of interstitial lung diseases . We describe the case of a girl with recurrent bacterial pneumonia and progressive interstitial fibrosis affecting the right lung . Morphologic evaluation of the lung biopsy showed structural changes of the vessel walls suggesting pulmonary hypertension . The echocardiogram showed the presence of centripetal blood flow in the right pulmonary artery from the periphery of the lung to the heart . A selective right angiography demonstrated the presence of pulmonary venous obstruction at the veno-atrial junction, successfully treated by endovascular stent implantation during cardiac catheterization . Comput Med Imaging Graph, 2002 Sep-Oct, 26(5), 327 - 32 Diffusion MRI in Rasmussen's encephalitis, herpes simplex encephalitis, and bacterial meningoencephalitis; Sener RN; Three patients with Rasmussen's encephalitis, herpes simplex type 1 encephalitis, and bacterial meningoencephalitis are included in this study . Echo-planar diffusion MRI was acquired with the trace protocol at 1.5 T . b= 1000 s/mm(2) images, and apparent diffusion coefficient (ADC) maps were studied with respect to lesion identification . ADC values were also studied, and compared to those of 25 normals . In Rasmussen's encephalitis b= 1000 s/mm(2) images were uninformative while ADC maps had superior information . In herpes simplex type 1 encephalitis both b= 1000 s/mm(2) images, and ADC maps had diagnostic information . In meningoencephalitis b= 1000 s/mm(2) images had superior information, especially with respect to early cerebritis while ADC maps were negative . In conclusion, diffusion MRI provided useful imaging data on different types of encephalitis, either on b= 1000 s/mm(2) images or on ADC maps, or on both. Mol Reprod Dev, 2002 Oct, 63(2), 161 - 7 Position-independent and tissue-specific expression of porcine whey acidic protein gene from a bacterial artificial chromosome in transgenic mice; Rival-Gervier S et al.; Silencing of transgenes is a frequent event after the random integration of foreign DNA in the host genome following microinjection . Long genomic fragments are expected to contain all the regulatory elements necessary to induce an appropriate expression of transgenes . A bacterial artificial chromosome containing the porcine wap gene with approximately 145 and 5 kb of 5'- and 3'-flanking sequences, respectively, was microinjected into fertilized mouse ovocytes . In the six transgenic lines studied, expression was strictly specific to the mammary gland of lactating animals and was position-independent . Levels of exogenous porcine wap mRNA per copy compared favorably with the porcine wap mRNA yield in the mammary gland of a 9-day lactating pig . These findings suggest that this insert contained most if not all of the cis-acting elements involved in the full specific expression of the porcine wap gene . These elements constitute good candidates for directing the optimized expression of protein recombinant-encoding genes in the mammary gland of lactating animals . Clin Infect Dis, 2002 Sep 15, 35(6), 678 - 83 Epub 2002 Aug 23. Prospective evaluation of a model of prediction of invasive bacterial infection risk among children with cancer, fever, and neutropenia; Santolaya ME et al.; A risk prediction model for invasive bacterial infection (IBI) was prospectively evaluated among children presenting with cancer, fever, and neutropenia . The model incorporated assessment of 5 previously identified risk factors: serum level of C-reactive protein (CRP) >/=90 mg/L, hypotension, identification of relapse of leukemia as the cancer type, platelet count of </=50,000 platelets/mm(3), and recent receipt of chemotherapy {16} . Children were uniformly evaluated at enrollment and were classified as having high or low risk for IBI according to a model that considers the number and type of variables present . Of the 263 febrile episodes evaluated during a 17-month period, 140 (53%) were in IBI-positive children . The sensitivity, specificity, and positive and negative predictive values of the model were 92%, 76%, 82%, and 90%, respectively . Identification of these 5 risk factors during the first 24 h of hospitalization was helpful in discriminating between children with a high or low risk for IBI. Anal Biochem, 2002 Aug 15, 307(2), 322 - 9 Fluorescent assay for polymerization of purified bacterial FtsZ cell-division protein; Trusca D et al.; Septum formation in Escherichia coli is a complex cascade of interactions among cell-division proteins . The tubulin-like FtsZ division protein localizes to the division site and serves a cytoskeletal role during septum formation . A novel fluorescent-based 96-well format filter assay has been developed to measure the polymerization of FtsZ . A mixture of monomers and aggregates (38 to approximately 200 KDa in range) of purified wild-type FtsZ and a fluorescently tagged derivative of FtsZ protein in stoichiometric ratio passes through a 0.2-microm filter membrane, while polymerized FtsZ is retained on the filter . Addition of the SulA protein to the assay leads to rapid disassembly of existing FtsZ polymers, demonstrating its natural regulatory effect on FtsZ under the assay conditions . This assay is sensitive (requiring 2 microM FtsZ or less) and facilitates high-throughput screening of factors affecting FtsZ polymerization. Biochem J, 2002 Dec 1, 368(Pt 2), 483 - 94 The first biantennary bacterial secondary cell wall polymer and its influence on S-layer glycoprotein assembly; Steindl C et al.; The cell surface of Aneurinibacillus thermoaerophilus DSM 10155 is covered with a square surface (S)-layer glycoprotein lattice . This S-layer glycoprotein, which was extracted with aqueous buffers after a freeze-thaw cycle of the bacterial cells, is the only completely water-soluble S-layer glycoprotein to be reported to date . The purified S-layer glycoprotein preparation had an overall carbohydrate content of 19% . Detailed chemical investigations indicated that the S-layer O-glycans of previously established structure accounted for 13% of total glycosylation . The remainder could be attributed to a peptidoglycan-associated secondary cell wall polymer . Structure analysis was performed using purified secondary cell wall polymer-peptidoglycan complexes . NMR spectroscopy revealed the first biantennary secondary cell wall polymer from the domain Bacteria, with the structure alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->4)-beta-L-Gal p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->4)-beta-L-Gal p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->4)-{alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->4)-beta-L-Gal p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->4)-beta-L-Gal p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)}-beta-L-Man p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->O)-PO(2)(-)-O-PO(2)(-)-(O-->6)-MurNAc- (where MurNAc is N -acetylmuramic acid) . The neutral polysaccharide is linked via a pyrophosphate bond to the C-6 atom of every fourth N -acetylmuramic acid residue, in average, of the A1gamma-type peptidoglycan . In vivo, the biantennary polymer anchored the S-layer glycoprotein very effectively to the cell wall, probably due to the doubling of motifs for a proposed lectin-like binding between the polymer and the N-terminus of the S-layer protein . When the cellular support was removed during S-layer glycoprotein isolation, the co-purified polymer mediated the solubility of the S-layer glycoprotein in vitro . Initial crystallization experiments performed with the soluble S-layer glycoprotein revealed that the assembly property could be restored upon dissociation of the polymer by the addition of poly(ethylene glycols) . The formed two-dimensional crystalline S-layer self-assembly products exhibited the same lattice symmetry as observed on intact bacterial cells. Hepatology, 2002 Sep, 36(3), 679 - 86 Bacterial motif DNA as an adjuvant for the breakdown of immune self-tolerance to pyruvate dehydrogenase complex; Jones DE et al.; Bacterial DNA containing unmethylated CpG dinucleotide motifs is immunostimulatory to mammals, skewing CD4(+) T-cell responses toward the Th1 phenotype . Autoreactive T-cell responses seen in primary biliary cirrhosis (PBC) are typically of the Th1 phenotype, raising the possibility that bacterial DNA might play a role in the generation of pathologic autoimmunity . We therefore studied the effects of CpG motif-containing oligodeoxynucleotides (ODN) on responses to pyruvate dehydrogenase complex (PDC, the autoantigen in PBC) in a murine model . Sensitization of SJL/J mice with non-self-PDC has been shown to result in induction of autoreactive T-cell responses to PDC sharing characteristics with those seen in patients with PBC . Administration of CpG ODN to SJL/J mice at the time of sensitization with PDC resulted in a significant skewing of splenic T-cell response to self-PDC, with significant augmentation of the Th1 cytokine response (interleukin {IL} 2 and interferon {IFN} gamma) and reduction of the Th2 response (IL-4 and IL-10) . In fact, CpG ODN seemed to be more effective at biasing the response phenotype and as effective at inducing liver histologic change as complete Freund's adjuvant (CFA), the standard adjuvant used for induction of Th1 responses in murine autoimmune and infectious immunity models . In conclusion, our findings raise the possibility that bacteria play a role in the development of autoimmunity (in PBC at least) through the potential of their DNA to shift the T-cell responses toward the phenotype associated with autoimmune damage . Moreover, this study suggests caution in the therapeutic use of CpG ODN as vaccine adjuvants. J Infect Dis, 2002 Sep 15, 186(6), 769 - 73 Epub 2002 Aug 28. Association of hypercoagulable states and increased platelet adhesion and aggregation with bacterial colonization of intravenous catheters; Musher D et al.; Bacterial adherence to intravenous catheters may be mediated, in part, by adherence to coagulation proteins and platelets . The possibility that catheter infection is associated with gene polymorphisms that cause hypercoagulability or increased platelet stickiness was examined . Among patients with infected catheters, there was no increase in the frequency of polymorphisms that increase coagulability, including factor V Leiden R506G, factor II (prothrombin) G20210A, and methylenetetrahydrofolate reductase C677T, compared with control subjects . The incidence of polymorphisms of the platelet beta(3) integrin among patients with infected catheters was also similar to that among control subjects . The C/D heterozygote of the variable number tandem repeat polymorphism and the C/T heterozygote of the KO polymorphism of glycoprotein Ibalpha were more frequent among patients with infected catheters than they were among control subjects . In a small proportion of patients, a genetic predisposition to platelet stickiness may be associated with infection of intravenous catheters, but in the majority, a recognized genetic tendency to hypercoagulability or platelet stickiness does not underlie infection. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai), 2002 Sep, 34(5), 533 - 43 Methods for structural and functional analysis of an RNA hexamer of bacterial virus phi29 DNA packaging motor; Guo PX; During multiplication and maturation, the lengthy genomic DNA of dsDNA viruses is translocated with remarkable velocity into a limited space within the procapsid and packaged to crystalline density . A viral DNA-packaging motor accomplishes this energy consuming motion task . An RNA molecule of bacterial virus phi29 has been found to be a vital component of the DNA-packaging motor . Six pRNAs form a hexagonal complex to gear the DNA translocating machine using a mechanism similar to the driving of a bolt with a hex nut . Sequential action of six RNA molecules to drive the motor is similar to the consecutive firing of six cylinders of a car engine . This article reviews the structure of pRNA to demonstrate that its structure plays a vital role in its function, and focuses on methods and unique approaches that lead to the elucidation of pRNA structure. EMBO J, 2002 Sep 2, 21(17), 4470 - 9 Dissociation of the dimeric SecA ATPase during protein translocation across the bacterial membrane; Or E et al.; The ATPase SecA mediates post-translational translocation of precursor proteins through the SecYEG channel of the bacterial inner membrane . We show that SecA, up to now considered to be a stable dimer, is actually in equilibrium with a small fraction of monomers . In the presence of membranes containing acidic phospholipids or in certain detergents, SecA completely dissociates into monomers . A synthetic signal peptide also affects dissociation into monomers . In addition, conversion into the monomeric state can be achieved by mutating a small number of residues in a dimeric and fully functional SecA fragment . This monomeric SecA fragment still maintains strong binding to SecYEG in the membrane as well as significant in vitro translocation activity . Together, the data suggest that the SecA dimer dissociates during protein translocation . Since SecA contains all characteristic motifs of a certain class of monomeric helicases, and since mutations in residues shared with the helicases abolish its translocation activity, SecA may function in a similar manner. Mol Divers, 2000, 5(3), 117 - 26 Identification of peptides that neutralize bacterial endotoxins using beta-hairpin conformationally restricted libraries; Gonzalez-Navarro H et al.; Bacterial endotoxins are the major mediator of septic shock; therefore, endotoxin-neutralizing molecules could have biomedical applications . The septic shock cascade relies in a series of molecular recognition processes . The large contact-surface described for the interacting macromolecules, in most cases, prevents the identification of small molecules that could modulate such recognition events . Here we report on a beta-hairpin conformationally restricted combinatorial library that has been generated and screened towards the identification of new peptides that neutralize bacterial endotoxins . Starting with a de novo designed linear peptide that shows a beta-hairpin structure population of around 30%, (Ramirez-Alvarado, M., Blanco, F . J . and Serrano, L . Nat . Struc . Biol., 7, 604-612 (1996)), we selected four positions to build up a combinatorial library of 20(4) sequences . Deconvolution of the library reduced such a sequence complexity to 8 defined sequences . The newly identified peptides have a biological activity equivalent to that reported for peptides derived from natural endotoxin-binding proteins. Proc Natl Acad Sci U S A, 2002 Sep 17, 99(19), 12415 - 20 Epub 2002 Aug 26. Cloning the vaccinia virus genome as a bacterial artificial chromosome in Escherichia coli and recovery of infectious virus in mammalian cells; Domi A et al.; The ability to manipulate the vaccinia virus (VAC) genome, as a plasmid in bacteria, would greatly facilitate genetic studies and provide a powerful alternative method of making recombinant viruses . VAC, like other poxviruses, has a linear, double-stranded DNA genome with covalently closed hairpin ends that are resolved from transient head-to-head and tail-to-tail concatemers during replication in the cytoplasm of infected cells . Our strategy to construct a nearly 200,000-bp VAC-bacterial artificial chromosome (BAC) was based on circularization of head-to-tail concatemers of VAC DNA . Cells were infected with a recombinant VAC containing inserted sequences for plasmid replication and maintenance in Escherichia coli; DNA concatemer resolution was inhibited leading to formation and accumulation of head-to-tail concatemers, in addition to the usual head-to-head and tail-to-tail forms; the concatemers were circularized by homologous or Cre-loxP-mediated recombination; and E . coli were transformed with DNA from the infected cell lysates . Stable plasmids containing the entire VAC genome, with an intact concatemer junction sequence, were identified . Rescue of infectious VAC was consistently achieved by transfecting the VAC-BAC plasmids into mammalian cells that were infected with a helper nonreplicating fowlpox virus. Int J STD AIDS, 2002 Aug, 13(8), 559 - 63 Diagnosis of bacterial vaginosis on self-collected vaginal tampon specimens; Sturm PD et al.; A vaginal tampon specimen was previously shown to be suitable for the molecular diagnosis of non-ulcerative sexually transmitted infections (STIs) . Different tampon fluid preparations were evaluated for the diagnosis of bacterial vaginosis (BV) . Women with pregnancy related problems were enrolled . Two observers evaluated the different tampon fluid preparations and vaginal smears collected during speculum examination using the Nugent score . Using the Amsel criteria, 21% of the 84 women enrolled were diagnosed with BV . Results of the tampon fluid preparations and vaginal smears showed excellent agreement for both observers (Spearman >0.80) . The overall sensitivity and specificity was 91.7% (95% CI: 81.6-96.5) and 79.3% (95% CI: 67.2-87.8), respectively, using the Amsel criteria as reference standard . The tampon provides a specimen for the combined diagnosis of non-ulcerative STIs and BV . This non-invasively collected specimen may facilitate self-initiated testing and population-based studies as well as longitudinal studies that are necessary to gain insight in the epidemiology of BV related to STIs and HIV. Clin Sci (Lond), 2002 Aug, 103 Suppl 48, 90S - 93S Characterization of PgPepO, a bacterial homologue of endothelin-converting enzyme-1; Carson JA et al.; PgPepO is a homologue of endothelin-converting enzyme-1 (ECE-1), with which it shares 31% identity . PgPepO was isolated from the periodontal pathogen Porphyromonas gingivalis . Recent studies have suggested a link between periodontal and cardiovascular disease, and several groups have suggested that bacterial and viral infections may contribute to the latter . P . gingivalis possesses the ability to invade, and multiply within, aortic endothelial cells and has been localized to atherosclerotic plaques . PgPepO was expressed and purified to homogeneity and we have begun detailed functional analysis, in terms of substrate preference and inhibitor specificity, in order to provide active-site comparisons with other members of the neprilysin (NEP)/ECE family . PgPepO possesses similar substrate specificity to ECE-1 and has been shown to cleave big endothelin-1 (big ET-1), big ET-2 and big ET-3, converting the substrates into their respective mature endothelin peptides . Substance P, angiotensin I, angiotensin II and neurotensin are all cleaved at multiple sites by PgPepO and the kinetics of these reactions have been compared . The potent vasoconstrictor urotensin II is not hydrolysed by PgPepO . Cleavage of bradykinin by PgPepO occurs at the Pro(7)-Phe(8) bond and is inhibited by the NEP and ECE-1 inhibitor phosphoramidon in a pH-dependent fashion (IC(50) =10 microM at pH 7.0) but not by thiorphan, an NEP-specific inhibitor . PgPepO activity is completely inhibited by EDTA . Characterization of this enzyme is important in elucidating possible links between periodontal pathogens and cardiovascular disorders such as atherosclerosis, and provides an opportunity to gain structural information on a bacterial protein with striking similarity to human ECE-1. Prog Urol, 2002 Jun, 12(3), 479 - 81 {Focal bacterial nephritis: diagnosis and treatment}; Ameur A et al.; Focal bacterial nephritis or lobar nephronia represents an acute localized non-liquefactive infection of the kidney caused by bacterial infection . This is an uncommon form of pyelonephritis that can affect both adults and children . Imaging techniques, particularly CT scan, are necessary for diagnosis and to distinguish it from other conditions (abscess or renal masses) that require a different treatment . The authors describe a case of acute lobar nephronia in a 24-year-old man. Proc Natl Acad Sci U S A, 2002 Sep 3, 99(18), 11605 - 10 Epub 2002 Aug 20. Reactions of cysteines substituted in the amphipathic N-terminal tail of a bacterial potassium channel with hydrophilic and hydrophobic maleimides; Li J et al.; Single cysteine-substitution mutants of KcsA, a K(+) channel from Streptomyces lividans, were expressed in Escherichia coli, and inner membranes were isolated . The rate constants for the reactions of these cysteines with three maleimides of increasing hydrophobicity, 4-(N-maleimido)phenyltrimethylammonium, N-phenylmaleimide, and N-(1-pyrenyl)maleimide, were determined by back titration of the remaining cysteines with methoxypolyethylene glycol-2-pyridine disulfide (M(r) 3,000) and quantitation of the fraction of gel-shifted KcsA as a function of reaction time . The patterns of the rate constants for the reactions of all three reagents with eight consecutive cysteines in the partially lipid-immersed amphipathic N-terminal tail helix were the same, with cysteines on the hydrophilic side of the helix reacting faster than Cys on the hydrophobic side . The results are consistent with the tail helix lying with its long axis in the lipid-water interface and with the orientation of the helix fluctuating around this axis . The patterns of the rate constants for the three reagents were similar to the pattern of the probabilities that the substituted cysteines were exposed to water, based on the sum of the free energies of transfer from water to octanol of all of the residues exposed to lipid in each orientation of the helix. J Biol Chem, 2002 Oct 25, 277(43), 40567 - 74 Epub 2002 Aug 19. Shiga-like toxin inhibition of FLICE-like inhibitory protein expression sensitizes endothelial cells to bacterial lipopolysaccharide-induced apoptosis; Erwert RD et al.; Shiga-like toxin (SLT) has been implicated in the pathogenesis of hemolytic uremic syndrome and its attendant endothelial cell (EC) injury . Key serotypes of Escherichia coli produce SLT-1 in addition to another highly pro-inflammatory molecule, lipopolysaccharide (LPS) . It has previously been established that SLT-1 induces EC apoptosis and that LPS enhances this effect . LPS alone has no affect on human EC viability, and the mechanism for this enhancement remains unknown . In the present report, we demonstrate that SLT-1 sensitizes EC to LPS-induced apoptosis . Pretreatment with SLT-1 sensitized EC to LPS-induced apoptosis, whereas pretreatment with LPS did not influence SLT-1-induced apoptosis . SLT-1 exposure resulted in decreased expression of FLICE-like inhibitory protein (FLIP), an anti-apoptotic protein that has previously been shown to block LPS-induced apoptosis . This SLT-1-mediated decrease in FLIP expression preceded the onset of apoptosis elicited by SLT-1 alone or in combination with LPS . SLT-1-mediated decrements in FLIP expression correlated in a dose- and time-dependent manner with sensitization to LPS-induced apoptosis . Finally, transient or stable overexpression of FLIP protected against LPS enhancement of SLT-1-induced apoptosis, and this protection corresponded with sustained expression of FLIP . Together, these data suggest that SLT-1 sensitizes EC to LPS-induced apoptosis by inhibiting FLIP expression. Zhonghua Nei Ke Za Zhi, 2002 Jul, 41(7), 459 - 61 Small bowel bacterial overgrowth and endotoxemia in cirrhosis; Wang J et al.; OBJECTIVE: To investigate the incidence of the small bowel bacterial overgrowth in cirrhotics and analyze the correlation among small bowel bacterial overgrowth (SBBO), plasma endotoxin level and plasma interleukin-2(IL-2), interleukin-6(IL-6), or tumor necrosis factor alpha(TNF-alpha) level . METHODS: Small bowel bacterial overgrowth in 71 cirrhotics was test by glucose hydrogen breath test (GHBT); plasma endotoxin in cirrhotics was measured with limilus lysate test; and plasma cytokine (IL-2, IL-6 and TNF-alpha) was measured with a solid-phase enzyme-linked immunosorbent assay; the incidence of SBBO in those 71 cirrhotics was investigated and the correlation between plasma endotoxin level and plasma IL-2, IL-6, or TNF-alpha level was analysed . RESULTS: (1)Positive GHBT were observed in 18 of 71 icrrhotics(25.3%); (2) Plasma endotoxin, IL-2, IL-6 and TNF-alpha levels were significantly higher in those cirrhotic patients with positive GHBT than in those with negative GHBT {(0.715 +/- 0.229) Eu/L versus (0.379 +/- 0.223) Eu/L, (19.15 +/- 4.60) ng/L versus (9.41 +/- 6.69) ng/L, (93.29 +/- 27.37) ng/L versus (53.22 +/- 28.31) ng/L, (42.18 +/- 16.91) ng/L versus (27.72 +/- 17.06) ng/L, respectively; P < 0.01}; (3) A significant correlation was observed between the level of plasma endotoxin and the level of plasma IL-2(r = 0.894, P < 0.001), IL-6(r = 0.857, P < 0.001) or TNF-alpha( r = 0.845,P < 0.001)in cirrhotics; CONCLUSIONS: (1)Plasma endotoxin, IL-2, IL-6,and TNF-alpha levels are increased in cirrhotic patients with SBBO, which suggests SBBO in cirrhotics may exasperated endotoxeamia;(2)Plasma endotoxin level in cirrhotics may stimulate some kinds of immune activated cells to produce IL-2, IL-6 and TNF-alpha, which may deteriorated cirrhosis or the complications. Nephron, 2002 Sep, 92(1), 213 - 5 Acute focal bacterial nephritis: report of four cases; Montejo M et al.; Focal acute bacterial nephritis is a localized bacterial infection of the kidney presenting as an inflammatory mass not containing drainable pus . The further distinction between acute focal bacterial nephritis and other renal masses is aided by the appropriate use of renal sonography and computed tomography . We report 4 cases with this entity . Proc Natl Acad Sci U S A, 2002 Sep 3, 99(18), 11611 - 5 Epub 2002 Aug 19. Dynamic and clustering model of bacterial chemotaxis receptors: structural basis for signaling and high sensitivity; Kim SH et al.; Bacterial chemotaxis receptors can detect a small concentration gradient of attractants and repellents in the environment over a wide range of background concentration . The clustering of these receptors to form patches observed in vivo and in vitro has been suspected as a reason for the high sensitivity, and such wide dynamic range is thought to be due to the resetting of the receptor sensitivity threshold by methylation/demethylation of the receptors . However, the mechanisms by which such high sensitivity is achieved and how the methylation/demethylation resets the sensitivity are not well understood . A molecular modeling of an intact bacterial chemotaxis receptor based on the crystal structures of a cytoplasmic domain and a periplasmic domain suggests an interesting clustering of three dimeric receptors and a two-dimensional, close-packed lattice formation of the clusters, where each receptor dimer contacts two other receptor dimers at the cytoplasmic domain and two yet different receptor dimers at the periplasmic domain . This interconnection of the receptors to form a patch of receptor clusters suggests a structural basis for the high sensitivity of the bacterial chemotaxis receptors . Furthermore, we present crystallographic data suggesting that, in contrast to most molecular signaling by conformational changes and/or oligomerization of the signaling molecules, the changes in dynamic property of the receptors on ligand binding or methylation may be the language of the signaling by the chemotaxis receptors . Taken together, the changes of the dynamic property of one receptor propagating mechanically to many others in the receptor patch provides a plausible, simple mechanism for the high sensitivity and the dynamic range of the receptors. Eur J Clin Pharmacol, 2002 Aug, 58(5), 321 - 2 Epub 2002 Jun 25. Bacterial pneumonia can increase serum concentration of clozapine; Raaska K et al.; Concentrations of serum clozapine, C-reactive protein (CRP) and alpha1 acid glycoprotein were greatly increased during a bacterial pneumonia in a 53-year-old woman . As the pneumonia subsided, and CRP and alpha1 acid glycoprotein normalised, serum clozapine concentration also decreased to the previous level . An increased serum clozapine and a lowered N-desmethylclozapine to clozapine ratio during the infection suggest a decreased cytochrome P(450) (CYP)1A2 activity . Cytokine-mediated CYP1A2 suppression is discussed. J Biol Chem, 2002 Nov 1, 277(44), 41489 - 96 Epub 2002 Aug 14. Molecular cloning of a novel chaperone-like protein induced by rhabdovirus infection with sequence similarity to the bacterial extracellular solute-binding protein family 5; Cho WJ et al.; Previously we demonstrated that a novel stress protein is induced in fish cells by the infection of a fish rhabdovirus (Cho W . J., Cha, S . J., Do, J . W., Choi, J . Y., Lee, J . Y., Jeong, C . S., Cho, K . J., Choi, W . S., Kang, H . S., Kim, H . D., and Park, J . W . (1997) Biochem . Biophys . Res . Commun . 233, 316-319) . In this paper, we present the molecular cloning and characterization of a gene encoding this protein named virus-inducible stress protein (VISP) . The VISP was purified partially by immunoprecipitation using a monoclonal antibody against the VISP and further purified by the electroelution from a SDS-PAGE gel . The protein was subjected to internal protein sequencing, and the sequence of three peptides was determined . Degenerate oligonucleotides based on the three peptide sequences were used to screen a cDNA library from rhabdovirus-infected CHSE-214 fish cells, and a cDNA of a 2193-bp open reading frame encoding the VISP with 730 amino acid residues (M(r) = 79.84) was identified . Whereas the nucleotide sequence of VISP shows no similarity with other genes in the GenBank(TM), the amino acid sequence of the VISP has similarity with the bacterial extracellular solute-binding protein family 5 (SBP_bac_5) that is proposed to have chaperone activity . Thus, we explored whether the VISP also had chaperone-like activity . Purified recombinant VISP expressed in Escherichia coli promoted the functional folding of alpha-glucosidase after urea denaturation and also prevented thermal aggregation of alcohol dehydrogenase . These results suggest that the VISP has amino acid sequence similarity with SBP_bac_5 and that it has chaperone activity that may play a role in virus infection. Proc Natl Acad Sci U S A, 2002 Sep 3, 99(18), 11772 - 7 Epub 2002 Aug 14. Identification of the binding sites of regulatory proteins in bacterial genomes; Li H et al.; We present an algorithm that extracts the binding sites (represented by position-specific weight matrices) for many different transcription factors from the regulatory regions of a genome, without the need for delineating groups of coregulated genes . The algorithm uses the fact that many DNA-binding proteins in bacteria bind to a bipartite motif with two short segments more conserved than the intervening region . It identifies all statistically significant patterns of the form W(1)N(x)W(2), where W(1) and W(2) are two short oligonucleotides separated by x arbitrary bases, and groups them into clusters of similar patterns . These clusters are then used to derive quantitative recognition profiles of putative regulatory proteins . For a given cluster, the algorithm finds the matching sequences plus the flanking regions in the genome and performs a multiple sequence alignment to derive position-specific weight matrices . We have analyzed the Escherichia coli genome with this algorithm and found approximately 1,500 significant patterns, which give rise to approximately 160 distinct position-specific weight matrices . A fraction of these matrices match the binding sites of one-third of the approximately 60 characterized transcription factors with high statistical significance . Many of the remaining matrices are likely to describe binding sites and regulons of uncharacterized transcription factors . The significance of these matrices was evaluated by their specificity, the location of the predicted sites, and the biological functions of the corresponding regulons, allowing us to suggest putative regulatory functions . The algorithm is efficient for analyzing newly sequenced bacterial genomes for which little is known about transcriptional regulation. Curr Opin Biotechnol, 2002 Jun, 13(3), 234 - 7 Disruption of bacterial quorum sensing by other organisms; Bauer WD et al.; Higher plants and algae produce compounds that mimic quorum sensing: signals used by bacteria to regulate the expression of many genes and behaviors . Similarly, various bacteria can stimulate, inhibit or inactivate quorum sensing in other bacteria . These discoveries offer new opportunities to manipulate bacterial quorum sensing in applications relevant to medicine, agriculture and the environment. Physiol Biochem Zool, 2002 May-Jun, 75(3), 273 - 82 The influence of bacterial lipopolysaccharide on the thermoregulation of the box turtle Terrapene carolina; do Amaral JP et al.; Ectotherms can adjust their thermoregulatory set points in response to bacterial infection; the result may be similar to endothermic fever . We examined the influence of dose on the set point of body temperature (T(b)) in Terrapene carolina . After acclimating postprandial turtles to 20 degrees C, we injected them with two doses of bacterial endotoxin (LPS; lipopolysaccharide from Escherichia coli), 0.0025 or 0.025 mg LPS/g nonshell body mass, or with reptilian saline (control group) . We placed the animals singly in linear thigmothermal gradients and recorded their T(b)'s for 48 h . The turtles showed dose-influenced thermal selection . Turtles injected with the high dose had T(b)'s significantly higher than control turtles, whereas low-dose turtles had T(b)'s significantly lower than control turtles . Also, there was a low daily effect on the T(b) of the turtles injected with the high dose . High-dose turtles had significantly higher T(b)'s than the control turtles during the first day but not during the second . Our results support the prediction of Romanovsky and Szekely that an infectious agent may elicit opposite thermoregulatory responses depending on quality and quantity of the agent and the host health status. Proc Natl Acad Sci U S A, 2002 Aug 20, 99(17), 10994 - 1001 Epub 2002 Aug 13. FAST CARS: engineering a laser spectroscopic technique for rapid identification of bacterial spores; Scully MO et al.; Airborne contaminants, e.g., bacterial spores, are usually analyzed by time-consuming microscopic, chemical, and biological assays . Current research into real-time laser spectroscopic detectors of such contaminants is based on e.g., resonance fluorescence . The present approach derives from recent experiments in which atoms and molecules are prepared by one (or more) coherent laser(s) and probed by another set of lasers . However, generating and using maximally coherent oscillation in macromolecules having an enormous number of degrees of freedom is challenging . In particular, the short dephasing times and rapid internal conversion rates are major obstacles . However, adiabatic fast passage techniques and the ability to generate combs of phase-coherent femtosecond pulses provide tools for the generation and utilization of maximal quantum coherence in large molecules and biopolymers . We call this technique FAST CARS (femtosecond adaptive spectroscopic techniques for coherent anti-Stokes Raman spectroscopy), and the present article proposes and analyses ways in which it could be used to rapidly identify preselected molecules in real time. Br Med Bull, 2002, 62, 113 - 23 Vaccines against human enteric bacterial pathogens; Dougan G et al.; The development of vaccines against enteric bacterial pathogens presents a challenge because of the large number of pathogens capable of causing disease and the requirement to induce immunity that is effective in the gut . A new generation of enteric vaccines based either on live or non-living antigens delivered orally or by injection are reaching the clinic in the early phases of evaluation . However, considerable technical barriers have to be overcome before these vaccines reach the general population. Cell Microbiol, 2002 Aug, 4(8), 541 - 56 Effect of Nramp1 on bacterial replication and on maturation of Mycobacterium avium-containing phagosomes in bone marrow-derived mouse macrophages; Frehel C et al.; Pathogenic mycobacteria prevent maturation of the phagosomes in which they reside inside macrophages and this is thought to be a major strategy allowing them to survive and multiply within macrophages . The molecular basis for this inhibition is only now beginning to emerge with the molecular characterization of the phagosome membrane enclosing these pathogens . We have used here several electron microscopy approaches in combination with counts of bacterial viability to analyse how expression of Nramp1 at the phagosomal membrane may influence survival of Mycobacterium avium and affect its ability to modulate the fusogenic properties of the phagosome in which it resides . The experiments were carried out in bone marrow-derived macrophages from wild-type 129sv (Nramp1(G169)) mice and from isogenic 129sv carrying a null mutation at Nramp1 (Nramp(1-/-)) following infection with a virulent strain of M . avium . We show here that Nramp1 expression has a bacteriostatic effect and that abrogation of Nramp1 restores the bacteria's capacity to replicate within macrophages . The combined analyses of the acquisition of endocytic contents markers delivered to early endosomes and/or lysosomes either prior to or after phagocytic uptake showed that in Nramp1-positive macrophages, M . avium was unable to prevent phagosome maturation and fusion with lysosomes but that in Nramp1-negative macrophages this capacity was restored . Several hypotheses are proposed to explain how Nramp1 could affect survival of M . avium . We also propose how the present observations could relate to the model according to which mycobacteria can prevent phagosome maturation by establishing a tight interaction with constituents of the phagosome membrane . Furthermore, these results show the importance of the choice of macrophages used as a model to study intracellular survival strategies of pathogens. Biochemistry, 2002 Aug 20, 41(33), 10426 - 38 Identification of A-minor tertiary interactions within a bacterial group I intron active site by 3-deazaadenosine interference mapping; Soukup JK et al.; The A-minor motifs appear to be the most ubiquitous helix packing elements within RNA tertiary structures . These motifs have been identified throughout the ribosome and almost every other tertiary-folded RNA for which structural information is available . These motifs utilize the packing of the donor adenosine's N1, N3, and/or 2'-OH against the 2'-OHs and minor groove edge of the acceptor base pair . The ability to identify biochemically which adenosines form A-minor motifs and which base pairs they contact is an important experimental objective . Toward this goal, we report the synthesis and transcriptional incorporation of 5'-O-(1-thio)-3-deazaadenosine triphosphate and its use in Nucleotide Analogue Interference Mapping (NAIM) and Nucleotide Analogue Interference Suppression (NAIS) . This analogue makes it possible for the first time to explore the functional importance of the N3 imino group of adenosine in RNA polymers . Interference analysis of the group I self-splicing introns from Tetrahymena and Azoarcus indicates that A-minor motifs are integral to the helix packing interactions that define the 5'-splice site of the intron . Specifically, Azoarcus A58 in the J4/5 region contacts the G.U wobble pair at the cleavage site in the P1 helix, and Azoarcus A167 in the J8/7 region contacts the C13-G37 base pair in the P2 helix . Both of these structural features are conserved between the eukaryotic and bacterial introns . These results suggest that nucleotide analogue interference patterns can identify and distinguish A-minor interactions in RNA tertiary structure, particularly the most prevalent type I and type II varieties . Furthermore, clustering of 3-deazaadenosine interferences is suggestive of A patches, in which a series of consecutive A-minor motifs mediate helix packing . Biochemical identification of these interactions may provide valuable constraints for RNA structure prediction. Biochemistry, 2002 Aug 20, 41(33), 10382 - 9 The purine nucleoside phosphorylase from Trichomonas vaginalis is a homologue of the bacterial enzyme; Munagala N et al.; Trichomonas vaginalis is a parasitic protozoan and the causative agent of trichomoniasis . Its primary purine salvage system, consisting of a purine nucleoside phosphorylase (PNP) and a purine nucleoside kinase, presents potential targets for designing selective inhibitors as antitrichomonial drugs because of lack of de novo synthesis of purine nucleotides in this organism . cDNA encoding T . vaginalis PNP was isolated by complementation of an Escherichia coli strain deficient in PNP and expressed, and the recombinant enzyme was purified to apparent homogeneity . It bears only 28% sequence identity with that of human PNP but 57% identity with the E . coli enzyme . Gel filtration showed the enzyme in a hexameric form, similar to the bacterial PNPs . Steady-state kinetic analysis of T . vaginalis PNP-catalyzed reactions gave K(m)'s of 31.5, 59.7, and 6.1 microM for inosine, guanosine, and adenosine in the nucleosidase reaction and 45.6, 35.9, and 12.3 microM for hypoxanthine, guanine, and adenine in the direction of nucleoside synthesis . This substrate specificity appears to be similar to that of bacterial PNPs . The catalytic efficiency of this enzyme with adenine as substrate is 58-fold higher than that with either hypoxanthine or guanine, representing a distinct disparity with the mammalian PNPs, which have negligible activity with either adenine or adenosine . The kinetic mechanism of T . vaginalis PNP-catalyzed reactions, determined by product inhibition and equilibrium isotope exchange, was by random binding of substrates (purine base and ribose 1-phosphate) with ordered release of the purine nucleoside first, followed by inorganic phosphate . Formycin A, an analogue of adenosine known as an inhibitor of E . coli PNP without any effect on mammalian PNPs, was shown to inhibit T . vaginalis PNP with a K(is) of 2.3 microM by competing with adenosine . T . vaginalis PNP thus belongs to the family of bacterial PNPs and constitutes a target for antitrichomonial chemotherapy. J Cereb Blood Flow Metab, 2002 Aug, 22(8), 988 - 96 Triptans reduce the inflammatory response in bacterial meningitis; Hoffmann O et al.; Severe headache and meningism provide clear evidence for the activation of trigeminal neurotransmission in meningitis . The authors assessed the antiinflammatory potential of 5HT1B/D/F receptor agonists (triptans), which inhibit the release of proinflammatory neuropeptides from perivascular nerve fibers . In a 6-hour rat model of pneumococcal meningitis, zolmitriptan and naratriptan reduced the influx of leukocytes into the cerebrospinal fluid, and attenuated the increase of regional cerebral blood flow . Elevated intracranial pressure as well as the brain water content at 6 hours was reduced by triptans . These effects were partially reversed by a specific 5HT1D as well as by a specific 5HT1B receptor antagonist . Meningitis caused a depletion of calcitonin gene-related peptide (CGRP) and substance P from meningeal nerve fibers, which was prevented by zolmitriptan and naratriptan . In line with these findings, patients with bacterial meningitis had significantly elevated CGRP levels in the cerebrospinal fluid . In a mouse model of pneumococcal meningitis, survival and clinical score at 24 hours were significantly improved by triptan treatment . The findings suggest that, besides mediating meningeal nociception, meningeal nerve fibers contribute to the inflammatory cascade in the early phase of bacterial meningitis . Adjunctive treatment with triptans may open a new therapeutic approach in the acute phase of bacterial meningitis. Int J STD AIDS, 2002 Jul, 13(7), 449 - 52 Bacterial vaginosis in climacteric and menopausal women; Taylor-Robinson D et al.; The objective was to determine how frequently an abnormal vaginal flora occurred in women attending a menopause clinic and whether any abnormality might be related to a particular risk factor . Women completed a questionnaire on their gynaecological, sexual and medical history . Whether they were perimenopausal or postmenopausal was determined on the basis of symptomatology, duration of amenorrhoea and on a follicle-stimulating hormone (FSH) assay when clinically indicated . A speculum examination of the vagina was undertaken, at which time a smear of vaginal secretion was Gram stained and the bacterial flora graded as follows: grade 1, normal; grade 2, intermediate, and grade 3, bacterial vaginosis (BV) . Of 100 women examined, 44 had grade 1 flora, 17 had grade 2 flora and 18 had BV . An apparent absence of, or very scanty, vaginal bacteria in which grading was not possible was found in 21 women . Women with BV had had more sexual partners than the others, but otherwise there were no discernible factors associated with the occurrence of BV . Women with vaginal atrophy were more likely to have an apparent absence of vaginal bacteria, but a few had BV. J Mol Diagn, 2002 Aug, 4(3), 144 - 9 Rapid detection of IgH/BCL2 rearrangement in follicular lymphoma by interphase fluorescence in situ hybridization with bacterial artificial chromosome probes; Jiang F et al.; Follicular lymphomas (FLs) can be difficult to diagnose on aspirated specimens since the architectural pattern is not present . FLs characteristically have rearrangements in the IgH and BCL2 genes resulting from the reciprocal t(14;18) (q32; q21) translocation . Because of the dispersed distribution of breakpoints, fluorescence in situ hybridization (FISH) using genomic probes that span or flank the breakpoints is ideal for detecting this rearrangement in fine-needle aspiration (FNA) biopsies . To develop a set of probes, a bacterial artificial chromosome library was screened and the clones were mapped by fiber FISH . The probes were produced by the direct incorporation of fluorochrome-labeled nucleotides . The colocalization base FISH assay was applied to Cytospin preparations from FNA biopsies of lymph nodes from 26 patients with FL and 10 patients without FL . In those with FL, the percentage of cells with at least one IgH/BCL2 fusion signal ranged from 22% to 100% (mean, 63%), which was statistically significantly higher than that in FL-negative samples (mean, 2.7%) . The probes demonstrated a significantly lower cutoff value (7%) in normal controls and effectively reduced the false-positive rate in FL-negative cases . These results were confirmed with fiber FISH assays on the same specimens . This interphase FISH assay is rapid and reliable for detecting rearrangements in the IGH/BCL2 gene, thereby aiding in the diagnosis of FL on FNA biopsy specimens. Nature, 2002 Aug 8, 418(6898), 662 - 5 Three-dimensional structure of the bacterial protein-translocation complex SecYEG; Breyton C et al.; Transport and membrane integration of polypeptides is carried out by specific protein complexes in the membranes of all living cells . The Sec transport path provides an essential and ubiquitous route for protein translocation . In the bacterial cytoplasmic membrane, the channel is formed by oligomers of a heterotrimeric membrane protein complex consisting of subunits SecY, SecE and SecG . In the endoplasmic reticulum membrane, the channel is formed from the related Sec61 complex . Here we report the structure of the Escherichia coli SecYEG assembly at an in-plane resolution of 8 A . The three-dimensional map, calculated from two-dimensional SecYEG crystals, reveals a sandwich of two membranes interacting through the extensive cytoplasmic domains . Each membrane is composed of dimers of SecYEG . The monomeric complex contains 15 transmembrane helices . In the centre of the dimer we observe a 16 x 25 A cavity closed on the periplasmic side by two highly tilted transmembrane helices . This may represent the closed state of the protein-conducting channel. Proc Natl Acad Sci U S A, 2002 Aug 20, 99(17), 11055 - 60 Epub 2002 Aug 07. Interactions between lipids and bacterial reaction centers determined by protein crystallography; Camara-Artigas A et al.; The structure of the reaction center from Rhodobacter sphaeroides has been solved by using x-ray diffraction at a 2.55-A resolution limit . Three lipid molecules that lie on the surface of the protein are resolved in the electron density maps . In addition to a cardiolipin that has previously been reported {McAuley, K . E., Fyfe, P . K., Ridge, J . P., Isaacs, N . W., Cogdell, R . J . & Jones, M . R . (1999) Proc . Natl . Acad . Sci . USA 96, 14706-14711}, two other major lipids of the cell membrane are found, a phosphatidylcholine and a glucosylgalactosyl diacylglycerol . The presence of these three lipids has been confirmed by laser mass spectroscopy . The lipids are located in the hydrophobic region of the protein surface and interact predominately with hydrophobic amino acids, in particular aromatic residues . Although the cardiolipin is over 15 A from the cofactors, the other two lipids are in close contact with the cofactors and may contribute to the difference in energetics for the two branches of cofactors that is primarily responsible for the asymmetry of electron transfer . The glycolipid is 3.5 A from the active bacteriochlorophyll monomer and shields this cofactor from the solvent in contrast to a much greater exposed surface evident for the inactive bacteriochlorophyll monomer . The phosphate atom of phosphatidylcholine is 6.5 A from the inactive bacteriopheophytin, and the associated electrostatic interactions may contribute to electron transfer rates involving this cofactor . Overall, the lipids span a distance of approximately 30 A, which is consistent with a bilayer-like arrangement suggesting the presence of an "inner shell" of lipids around membrane proteins that is critical for membrane function. Auris Nasus Larynx, 2002 Jul, 29(3), 241 - 5 Immunoreactivity for myeloperoxidase (MPO) in the vestibule after the injection of bacterial lipopolysaccharide into the middle ear; Watanabe K et al.; OBJECTIVE: In this study, the effect of endotoxin on the vestibule of the ear of guinea pigs was immunohistochemically examined . METHODS: Bacterial lipopolysaccharide was injected into the middle ear transtympanically . After 48 h, the animals were sacrificed by intracardiac perfusion of fixative; then, the temporal bones were removed and processed for immunohistochemical staining with the anti-myeloperoxidase antibody . RESULTS: Myeloperoxidase could be detected after 48 h in the sensory epithelium and the dark cell area . CONCLUSION: It is reported that radical oxygen species, which are cytotoxic, are detected under inflammatory conditions . Our results suggest that myeloperoxidase and reactive oxygen species are involved in vestibular dysfunction under inflammation. Theor Popul Biol, 2002 Jun, 61(4), 503 - 7 Preferential orientation of natural lambdoid prophages and bacterial chromosome organization; Campbell AM; All known lambdoid prophages of Escherichia coli have the same orientation with respect to direction of chromosomal replication . This includes 12 prophages that are replicated in one direction and five in the other . Among candidate explanations, the most amenable to experimental study is an effect on dif site function in assuring chromosomal segregation . This is but one of numerous examples of strand bias in the E . coli genome, all of which may interact with one another . Shock, 2002 Aug, 18(2), 148 - 51 The effects of pentoxifylline on bacterial translocation after intestinal obstruction; Kocdor MA et al.; Bacterial translocation (BT) occurs mainly in preseptic conditions such as intestinal obstruction, trauma, and burn, and the underlying mechanisms are still unclear . Pentoxifylline (PTX) is a derivative of methyl xanthine and has several beneficial effects in sepsis . We investigated the effects of PTX on a rat BT model . Simple intestinal obstruction (IO) was choosen to create high BT rates . Rats were divided in to five groups of 10 rats . Either 50 mg/kg PTX or placebo (3 mg/100 g saline) was administered subcutaneously following IO, either by single injection or twice with a 12-h interval . All rats were sacrificed 12 or 24 h after the procedure, and mesenteric lymph nodes (MLN), liver, and blood samples were obtained under aseptic conditions for bacterial cultures . The samples were obtained 12 h following IO in the first two groups, and the same samples were obtained 24 h after IO in last three groups . Groups IV and V were the PTX treatment groups . PTX was re-injected 12 h after IO only in group IV . As a result, BT rates in MLNs and liver were found to be significantly low, blood specimens remained sterile in PTX-pretreated and -treated rats, and BT rates were high in control groups and group V (once treatment late specimen group) . We conclude that simple intestinal obstruction causes BT, and PTX reduces BT in rats with IO during the first 12-h period if PTX is given once immediately following IO . PTX reduces BT during the first 24-h after IO provided that is injected twice with a 12-h interval . More experimental studies are need to explain the exact mechanism of this beneficial effect. Endocrine, 2002 Jun, 18(1), 13 - 20 Bacterial lipopolysaccharide-induced coordinate downregulation of arginine vasopressin receptor V3 and corticotropin-releasing factor receptor 1 messenger ribonucleic acids in the anterior pituitary of endotoxemic steers; Qahwash IM et al.; AVP and CRF are potent stimulators of pituitary ACTH secretion in cattle . Actions of AVP and CRF at the anterior pituitary are mediated by AVP receptor V3 (V3) and CRF receptor 1 (CRFR1) . The primary objective of these studies was to determine the effect of systemic inflammatory stress on V3 and CRFR1 mRNAs in the bovine anterior pituitary . Holstein steers (n = 20) were injected with 200 ng/kg bacterial lipopolysaccharide (LPS) and tissues collected 0, 2, 4, 12, and 24 h later . All animals responded to LPS administration with an increase in body temperature, plasma ACTH, and cortisol (p < 0.05) . Abundance of anterior pituitary V3 mRNA was decreased at 2, 4, and 12 h following LPS administration (p < 0.05) and returned to basal by 24 h . A similar temporal regulation of pituitary CRFR1 mRNA (p < 0.05), but not pituitary pro-opiomelanocortin (POMC) mRNA, was observed following LPS administration . Similar downregulation of CRFR1 mRNA was not observed in other brain regions following LPS administration (cerebellum, hypothalamus) . Our results indicate that V3 and CRFR1 mRNAs are coordinately downregulated in the anterior pituitary during systemic inflammatory stress . Decreased AVP and CRF receptor expression may help regulate the pituitary-adrenal response to stress. Oral Dis, 2002, 8 Suppl 2, 80 - 7 Oral fungal and bacterial infections in HIV-infected individuals: an overview in Africa; Hodgson TA et al.; Oral opportunistic infections developing secondary to human immunodeficiency virus (HIV) infection have been reported from the early days of the epidemic and have been classified by both the EC-Clearinghouse and the World Health Organisation (WHO) . Among the fungal infections, oral candidiasis, presenting in African HIV-infected patients has been sporadically documented . We review the literature with respect to candidal carriage, oral candidiasis prevalence and the predictive value of oral candidiasis for a diagnosis of underlying HIV disease in African HIV-infected patients . The use of oral candidiasis as a marker of disease progression, the species of yeasts isolated from the oral cavity in Africa and the resistance of the yeasts to antifungal agents and treatment regimens are discussed . Orofacial lesions as manifestations of the systemic mycoses are rarely seen in isolation and few cases are reported in the literature from Africa . In spite of the high incidence of noma, tuberculosis, chronic osteomyelitis and syphilis in Africa, surprisingly there have been very few reported cases of the oral manifestations of these diseases in HIV-positive individuals . Orofacial disease in HIV-infected patients is associated with marked morbidity, which is compounded by malnutrition . The authors indicate specific research areas, initially directed at the most effective management strategies, which would complete data in this important area. Kaohsiung J Med Sci, 2002 Apr, 18(4), 164 - 70 Improving Gram-stained reproducible result by further adding clue cells in diagnosing bacterial vaginosis; Lin DP et al.; The reproducibility of interpretation in diagnosing bacterial vaginosis may be enhanced by adding pus cells and clue cells into two different criteria, developed by Spiegel et al . and Nugent et al . The purpose of study was designed to find out which parameter was more reproducible . 100 patients were collected with the diagnosis of bacterial vaginosis as an experimental group, while the other 100 patients who were with routine Papanicolaou smears in gynecologic clinic the collected as a control group . Two slides, including the original and reproducible ones, were obtained from vaginal smears for each patient . Three technicians read the slides randomly by using two different criteria, plus pus cells and clue cells . This showed the agreement for clue cells is the best method regardless of experimental group or control group (Kappa values between 0.708 and 1.000) . The intra-observer agreement for the diagnosis of bacterial vaginosis by the method of Nugent et al . is superior to the method of Spiegel et al . Our data show the comparison of Amsel criteria versus Nugent criteria for the diagnosis of bacterial vaginosis with sensitivity of 88.9%, specificity of 55.4%, negative positive value of 62.1%, and positive predictive value of 85.8% . Moreover, our data also demonstrate the comparison of Amsel criteria versus the diagnosis either based on Nugent criteria or the presence of clue cells with sensitivity of 95.7%, specificity of 56.7%, negative positive value of 81.2%, and positive predictive value of 87.1% . The results demonstrate further adding score of the clue cells can enhance the reproducible diagnosis of bacterial vaginosis, which is superior to the methods of Nugent et al . and Spiegel et al. Otolaryngol Head Neck Surg, 2002 Jul, 127(1), 1 - 6 Short treatment durations for acute bacterial rhinosinusitis: Five days of gemifloxacin versus 7 days of gemifloxacin; Ferguson BJ et al.; OBJECTIVE: The primary objective of this study was to demonstrate the clinical and radiologic efficacy of 5 days compared with 7 days of gemifloxacin therapy in the treatment of acute bacterial rhinosinusitis (ABRS) . STUDY DESIGN: In this prospective, double-blind, multicenter, parallel-group study, adult patients presenting with ABRS were randomized to receive gemifloxacin 320 mg once daily for either 5 days (n = 218) or 7 days (n = 203) . RESULTS: For the primary efficacy end point, clinical response to therapy at follow-up, 5 days of therapy with gemifloxacin was as effective as 7 days of therapy (per-protocol population; treatment difference 0.44%; 95% confidence interval {CI}, -6.54 to 7.41) . Five and 7 days of treatment with gemifloxacin were well tolerated . CONCLUSION AND SIGNIFICANCE: The clinical efficacy of gemifloxacin 320 mg daily for 5 days is at least as good as the efficacy of gemifloxacin 320 mg daily for 7 days in the treatment of ABRS. Environ Microbiol, 2002 Aug, 4(8), 477 - 81 nifH gene diversity in the bacterial community associated with the rhizosphere of Molinia coerulea, an oligonitrophilic perennial grass; Hamelin J et al.; Rhizosphere associative dinitrogen fixation could be a valuable source of nitrogen in many nitrogen limited natural ecosystems, such as the rhizosphere of Molinia coerulea, a hemicryptophytic perennial grass naturally occurring in contrasted oligonitrophilic soils . The diversity of the dinitrogen-fixing bacteria associated with this environment was assessed by a cloning-sequencing approach on the nifH gene directly amplified from environmental DNA extracts . Seventy-seven randomly picked clones were analysed . One type of NifH sequence was dominant in both roots and surrounding soil, and represented 56% of all retrieved sequences . This cluster included previously described environmental clones and did not contain any NifH sequences similar to cultivated diazotrophs . The predominance of few NifH sequence types in the roots and the rhizosphere of Molinia coerulea indicate that the plant environment mediates a favourable niche for such dinitrogen-fixing bacteria. Biotechnol Prog, 2002 Jul-Aug, 18(4), 686 - 93 Nonlinear dynamics of regulation of bacterial trp operon: model analysis of integrated effects of repression, feedback inhibition, and attenuation; Xiu ZL et al.; The trp operon encodes the five genes for the enzymes required to convert chorismate to tryptophan, and its switching on and off is controlled by both feedback repression and attenuation in response to different levels of tryptophan in the cell . Repression of the operon occurs when tryptophan concentration is high, and attenuation fine-tunes the transcription level at a lower cellular concentration of tryptophan . An extended mathematical model is established in this study to describe the switching on and off of the trp operon by considering the integrated effects of repression and attenuation . The influences of cell growth rate on the biosynthesis of tryptophan, stability and dynamic behavior of the trp operon are investigated . Sustained oscillations of tryptophan levels are predicted from the regulated turning on and off of the trp operon . It is interesting to note that during such oscillations the regulation of transcription displays a kind of "on" and "off" state in terms of gene expression, indicating the existence of a genetic circuit or switch in the regulation of the trp operon . Time lags between transcription and translation are also predicted and may explain the occurrence of such oscillation phenomenon. Water Res, 2002 May, 36(10), 2561 - 70 Hydrogeochemical controls on the organic matter and bacterial ecology of a small freshwater wetland in the New Jersey Pine Barrens; Maurice PA et al.; This study investigated the effects of variable ground-water/surface-water exchange and photoinduced processes on longitudinal patterns in dissolved organic matter (DOM) and bacterial communities in a small first-order stream in the New Jersey Pine Barrens . DOM concentration, along with DOM weight average molecular weight (Mw) and absorptivity (epsilon280, an estimator of aromaticity), and bacterial cell counts all decreased from the stream and hyporheic zone into the shallow aquifer in a ground-water recharge zone . Further downstream, influx of ground water into the stream resulted in a lower Mw DOM pool and was accompanied by decreased cell counts . The observed effect of this ground-water discharge on bacterial numbers may be direct, if discharge temporarily dilutes cell counts, or indirect, if changes in DOM concentration and properties control the bacterial community . In either case, this study suggested the importance of considering ground-water-surface-water exchange in studies of longitudinal changes in the bacterial communities of streams. Dis Aquat Organ, 2002 Jun 21, 50(1), 1 - 8 Usefulness of dead shrimp specimens in studying the epidemiology of white spot syndrome virus (WSSV) and chronic bacterial infection; Mohan CV et al.; This paper describes the utility of dead shrimp samples in epidemiological investigations of the white spot syndrome virus (WSSV) and chronic bacterial infections . A longitudinal observational study was undertaken in shrimp farms in Kundapur, Karnataka, India, from September 1999 to April 2000 to identify risk factors associated with outbreaks of white spot disease (WSD) in cultured Penaeus monodon . As a part of the larger study, farmers were trained to collect and preserve dead and moribund shrimp (when observed) during the production cycle . At the end of the production cycle, 73 samples from 50 ponds had been collected for histopathology and 55 samples from 44 ponds for PCR . Intranuclear viral inclusion bodies diagnostic of WSSV infection were detected in dead samples from 32 ponds (64 %) . Samples of dead shrimp from 18 ponds (36%) showed no histopathological evidence of WSSV infection . However, of these, samples from 13 ponds (26%) showed clear evidence of shell, oral, enteric and systemic chronic inflammatory lesions (CIL) in the form of haemocytic nodules, typical of bacterial infection . Samples from 5 ponds (10%) were negative for both WSSV and CIL