J Biol Chem, 2003 May 2, 278(18), 16320 - 8 Epub 2003 Feb 24. A conserved structural motif at the N terminus of bacterial translation initiation factor IF2; Laursen BS et al.; The 18-kDa Domain I from the N-terminal region of translation initiation factor IF2 from Escherichia coli was expressed, purified, and structurally characterized using multidimensional NMR methods . Residues 2-50 were found to form a compact subdomain containing three short beta-strands and three alpha-helices, folded to form a betaalphaalphabetabetaalpha motif with the three helices packed on the same side of a small twisted beta-sheet . The hydrophobic amino acids in the core of the subdomain are conserved in a wide range of species, indicating that a similarly structured motif is present at the N terminus of IF2 in many of the bacteria . External to the compact 50-amino acid subdomain, residues 51-97 are less conserved and do not appear to form a regular structure, whereas residues 98-157 form a helix containing a repetitive sequence of mostly hydrophilic amino acids . Nitrogen-15 relaxation rate measurements provide evidence that the first 50 residues form a well ordered subdomain, whereas other regions of Domain I are significantly more mobile . The compact subdomain at the N terminus of IF2 shows structural homology to the tRNA anticodon stem contact fold domains of the methionyl-tRNA and glutaminyl-tRNA synthetases, and a similar fold is also found in the B5 domain of the phenylalanine-tRNA synthetase . The results of the present work will provide guidance for the design of future experiments directed toward understanding the functional roles of this widely conserved structural domain within IF2. Biochemistry, 2003 Mar 4, 42(8), 2412 - 21 Bacterial expression and characterization of a novel, soluble, calcium-binding, and calcium-activated human nucleotidase; Murphy DM et al.; A newly discovered human analogue of a bed bug apyrase, which we named hSCAN-1 for human soluble calcium-activated nucleotidase-1, was expressed in bacteria, refolded from inclusion bodies, purified, and characterized . This apyrase, which is distinct from the eNTPDases exemplified by the endothelial CD39 (NTPDase1) apyrase, is a 38 kDa monomeric enzyme capable of hydrolyzing a variety of nucleoside di- and triphosphates, but not monophosphates . Preferred substrates include GDP, UDP, and IDP, with a pH optimum for activity between 6 and 7 . The specific activity and substrate preference of the bacterially expressed enzyme closely mimic those of the enzyme expressed in mammalian COS cells, as well as the enzyme synthesized in an in vitro bacterial expression system . This suggests that glycosylation and other posttranslational modifications that do not occur in bacteria are not necessary for nucleotidase activity or proper folding of this human apyrase . hSCAN-1 absolutely requires Ca(2+), but not Mg(2+) or other divalent cations analyzed, for enzymatic activity . Surprisingly, the activity does not increase in a quasi-linear fashion at sub-millimolar Ca(2+) concentrations, as would be expected if Ca(2+) were only used as a cosubstrate for the nucleotide substrate, but rather follows a sigmoidal curve . The intrinsic fluorescence and difference absorption studies of hSCAN-1 in the absence of nucleotides revealed Ca(2+)-induced changes in the environment of tryptophan and tyrosine residues with half-saturation at about 90 microM Ca(2+) . NaCl increased the half-saturating Ca(2+) concentration needed for both structural changes detected by optical spectroscopy and enzymatic activation of hSCAN-1 detected by nucleotidase assay . These results suggest that Ca(2+) triggers a conformational change in hSCAN-1, converting the enzymatically inactive protein to the active enzyme, in addition to forming the metal-nucleotide substrate complex necessary for nucleotidase activity. Pediatr Surg Int, 2002 Dec, 18(8), 712 - 7 Epub 2002 Oct 26. Intraluminal casein model of necrotizing enterocolitis for assessment of mucosal destruction, bacterial translocation, and the effects of allopurinol and N-acetylcysteine; Koivusalo A et al.; An intraluminal casein model (ICM) of necrotizing enterocolitis (NEC) is able to produce small-bowel changes reminiscent of human NEC in neonatal animals . We studied bacterial translocation (BT) in NEC induced by using the ICM in neonatal piglets . We also studied whether allopurinol (AL) and N-acetylcysteine (NAC) have an effect on BT and mucosal changes in the ICM of NEC . Twenty-eight neonatal piglets were randomized into four groups . NEC was induced in 21 by injecting casein-d-gluconate into a loop of terminal ileum: group Cas (n = 7) had no premedication, in group Cas/AL (n = 7) intravenous (i.v.) Al (100 mg/kg), and in group Cas/NAC (n = 7) i.v . NAC (200 mg/kg) was given . Group Sham (n = 7) had the ileum injected with 0.9% saline with no premedication . Immediately after the injection a mesenteric lymph node (MLN) adjacent to the loop was harvested for quantitative aerobic bacterial culture; 4 h after the injection another MLN and samples of spleen, liver, kidney, and lung were harvested and cultured . Comparison of the incidence of samples with positive bacterial cultures and the number of colony-forming units (CFU) in samples was made between groups . The severity of NEC in the ileum was graded from 0 to 3 according to macroscopic and histologic findings . NEC changes in the bowel were most severe in Cas piglets, less severe in Cas/NAC piglets ( P < 0.5), and sham piglets had the least severe changes ( P < 0.05) . piglets with NEC changes in the ileum had a higher incidence of BT into the MLN than piglets without NEC changes ( P < 0.05), but the difference in CFU was not significant ( P > 0.05) . In Cas and Cas/NAC piglets a high incidence of BT into the MLN was noted as early at -5 min after casein injection . The incidence of BT into the MLN was significantly higher in Cas and Cas/NAC piglets than in Sham piglets ( P < 0.05), the difference in CFU being not significant ( P > 0.05) . BT in Cas/Al piglets was not significantly different from that of Cas piglets ( P > 0.05), but less than in Cas/NAC piglets ( P < 0.05) . Four hours after casein injection into the ileum there was significant BT into the MLN . Premedication with NAC was associated with less severe NEC changes, but neither NAC nor AL significantly affected BT. AIDS, 2003 Mar 7, 17(4), 575 - 84 Pattern of bacterial diseases in a cohort of HIV-1 infected adults receiving cotrimoxazole prophylaxis in Abidjan, Côte d'Ivoire; Anglaret X et al.; BACKGROUND: WHO/UNAIDS recommended that cotrimoxazole should be prescribed in Africa in HIV-infected adults with CD4 cell counts < 500 x 10 /l, while closely monitoring bacterial diseases in as many settings as possible . METHODS: Prospective cohort study, describing bacterial morbidity in adults receiving cotrimoxazole prophylaxis (960 mg daily) between April 1996 and June 2000 in Abidjan, Cote d'Ivoire . RESULTS: Four-hundred and forty-eight adults (median baseline CD4 cell count 251 x 10 /l) were followed for a median time of 26 months . The rates of overall bacterial diseases and of serious bacterial diseases with hospital admission were 36.8/100 person-years (PY) and 11.3/100 PY, respectively . Bacterial diseases were the first causes of hospital admissions, followed by non-specific enteritis (10.2/100 PY), acute unexplained fever (8.4/100 PY), and tuberculosis (3.6/100 PY) . Among serious bacterial diseases, the most frequent were enteritis (3.0/100 PY), invasive urogenital infections (2.5/100 PY), pneumonia (2.3/100 PY), bacteraemia with no focus (2.0/100 PY), upper respiratory tract infections (1.6/100 PY) and cutaneous infections (0.6/100 PY) . Compared with patients with baseline CD4+ cell counts >or= 200 x 10 /l, other patients had an adjusted hazard ratio of serious bacterial diseases of 3.05 (95% confidence interval, 2.00-4.67; < 0.001) . Seventy-five bacterial strains were isolated during serious episodes including 29 non-, 14, 12 spp, and 12 . DISCUSSION: Though with a medium-term rate half that of the short-term rate estimated under placebo before 1998 (26.1/100 PY), serious bacterial morbidity remains the first cause of hospital admission in adults receiving cotrimoxazole in this setting. Arch Dis Child, 2003 Mar, 88(3), 222 - 3 Bacterial infections, immune overload, and MMR vaccine . Measles, mumps, and rubella; Miller E et al.; Combined measles, mumps, and rubella (MMR) vaccine did not increase the risk of hospitalisation with invasive bacterial infection in the three months after vaccination; rather there was a protective effect . These results provide no support for the concept of "immunological overload" induced by multiple antigen vaccinations, nor calls for single antigen vaccines. Protein Expr Purif, 2003 Feb, 27(2), 279 - 92 A bicistronic expression system for bacterial production of authentic human interleukin-18; Kirkpatrick RB et al.; Interleukin-18 (IL-18) is activated and released from immune effector cells to stimulate acquired and innate immune responses involving T and natural killer (NK) cells . The release of IL-18 from mammalian cells is linked to its proteolytic activation by caspases including interleukin 1 converting enzyme (ICE) . The absence of a signal peptide sequence and the requirement for coupled activation and cellular release have presented challenges for the large-scale recombinant production of IL-18 . In this study, we have explored methods for the direct production of authentic human IL-18 toward the development of a large-scale production system . Expression of mature IL-18 directly in Escherichia coli with a methionine initiating codon leads to the production of MetIL-18 that is dramatically less potent in bioassays than IL-18 produced as a pro-peptide and activated in vitro . To produce an authentic IL-18, we have devised a bicistronic expression system for the coupled transcription and translation of ProIL-18 with caspase-1 (ICE) or caspase-4 (ICE-rel II, TX, ICH-2) . Mature IL-18 with an authentic N-terminus was produced and has a biological activity and potency comparable to that of in vitro processed mature IL-18 . Optimization of this system for the maximal production yields can be accomplished by modulating the temperature, to affect the rate of caspase activation and to favor the accumulation of ProIL-18, prior to its proteolytic processing by activated caspase . The effect of temperature is particularly profound for the caspase-4 co-expression process, enabling optimized production levels of over 150 mg/L in shake flasks at 25 degrees C . An alternative bicistronic expression design utilizing a precise ubiquitin IL-18 fusion, processed by co-expressed ubiquitinase, was also successfully used to generate fully active IL-18, thereby demonstrating that the pro-sequence of IL-18 is not required for recombinant IL-18 production . South Med J, 2002 Dec, 95(12), 1408 - 10 Bacterial contamination of paper currency; Pope TW et al.; One-dollar bills were collected from the general community in western Ohio to survey for bacterial contamination . Pathogenic or potentially pathogenic organisms were isolated from 94% of the bills . These results suggest a high rate of bacterial contamination of one-dollar bills. Indian J Exp Biol, 2002 Aug, 40(8), 924 - 9 A rapid hypochlorite method for extraction of polyhydroxy alkanoates from bacterial cells; Rawte T et al.; A new method has been standardized for extraction of polyhydroxy alkanoates from the bacteria, using sodium hypochlorite . This method is simple and quick as compared to the existing methods . Statistical analysis has proved the method to be reliable and reproducible. APMIS, 2002 Nov, 110(11), 811 - 8 An international study of the interobserver variation between interpretations of vaginal smear criteria of bacterial vaginosis; Forsum U et al.; An international workshop on vaginal smear-based diagnosis of bacterial vaginosis was organized where 13 investigators scoring 258 slides with smears from vaginal fluid . Interobserver reproducibility of interpretations of Nugent scores, Hay/Ison scores and wet smear scores for the diagnosis of bacterial vaginosis was shown to be high . Detailed analysis of individual scoring results however indicated that basic standards of quality control to ensure robust individual readings of slides must be adhered to. Proteins, 2003 Apr 1, 51(1), 85 - 95 BATMAS30: amino acid substitution matrix for alignment of bacterial transporters; Sutormin RA et al.; Aligned amino acid sequences of three functionally independent samples of transmembrane (TM) transport proteins have been analyzed . The concept of TM-kernel is proposed as the most probable transmembrane region of a sequence . The average amino acid composition of TM-kernels differs from the published amino acid composition of transmembrane segments . TM-kernels contain more alanines, glycines, and less polar, charged, and aromatic residues in contrast to non-TM-proteins . There are also differences between TM-kernels of bacterial and eukaryotic proteins . We have constructed amino acid substitution matrices for bacterial TM-kernels, named the BATMAS (BActerial Transmembrane MAtrix of Substitutions) series . In TM-kernels, polar and charged residues, as well as proline and tyrosine, are highly conserved, whereas there are more substitutions within the group of hydrophobic residues, in contrast to non-TM-proteins that have fewer, relatively more conserved, hydrophobic residues . These results demonstrate that alignment of transmembrane proteins should be based on at least two amino acid substitution matrices, one for loops (e.g., the BLOSUM series) and one for TM-segments (the BATMAS series), and the choice of the TM-matrix should be different for eukaryotic and bacterial proteins . Theor Appl Genet, 2003 Feb, 106(4), 765 - 70 Epub 2002 Oct 22. Identification of DNA markers of tobacco linked to bacterial wilt resistance; Nishi T et al.; Bacterial wilt caused by Ralstonia solanacearum is one of the most destructive soil-borne diseases in the world . Breeding resistant commercial varieties of tobacco is difficult because most donor candidates' resistance is controlled by polygenes . In this paper, we demonstrate the identification of useful DNA markers for bacterial wilt-resistant tobacco breeding . One hundred and seventeen markers were identified by the amplified fragment length polymorphism (AFLP) method between W6, a burley variety with resistance originating from a Japanese domestic variety, Hatano, and Michinoku 1, a commercial burley wilt-susceptible variety, using 3,072 primer combinations . These markers were analyzed in 125 doubled haploid lines, derived from F(1) hybrids between W6 and Michinoku 1, and a linkage map consisting of ten linkage groups was drawn . The resistance phenotype of each of these lines was investigated on the basis of the average of disease severity obtained from field trials over two growing cycles . Quantitative trait loci (QTL) analysis was performed on the marker phenotypes and the resistance phenotype of each line . One QTL for the bacterial wilt resistance of W6 and DNA markers associated with this QTL were identified on a linkage group consisting of 15 markers, 32 cM in length . This QTL explained more than 30% of the variance in resistance among these lines. J Mol Biol, 2003 Mar 7, 326(5), 1413 - 26 Visualizing RNA extrusion and DNA wrapping in transcription elongation complexes of bacterial and eukaryotic RNA polymerases; Rivetti C et al.; Transcription ternary complexes of Escherichia coli RNA polymerase and yeast RNA polymerase III have been analyzed by atomic force microscopy . Using the method of nucleotide omission and different DNA templates, E.coli RNAP has been stalled at position +24, +70 and +379 and RNAP III at position +377 from the starting site . Conformational analysis of E.coli RNAP elongation complexes reveals an average DNA compaction of 22nm and a DNA deformation compatible with approximately 180 degrees DNA wrapping against the enzyme . The extent of protein-DNA interaction attributed to wrapping, however, is less than that of corresponding open promoter complexes . DNA wrapping was also observed for RNAP III elongation complexes, which showed a DNA compaction of 30nm . When the RNA polymerases were stalled far from the promoter (+379 and +377), the growing RNA transcript was often visible and it was prevalently seen exiting from the enzyme on the opposite side relative to the smallest angle subtended by the upstream and downstream DNA arms . Surprisingly, we found that many complexes had a second RNAP, not involved in transcription, bound to the growing RNA of a ternary complex . DNA wrapping in the elongation complex suggests a possible mechanism by which the polymerase may overcome the physical barrier to transcription imposed by the nucleosomes. Surg Infect (Larchmt), 2000 Winter, 1(4), 265 - 72 Transcellular Transport Is not Required for Transmucosal Bacterial Passage across the Intestinal Membrane Ex Vivo; Nadler EP et al.; The mechanisms underlying the process of bacterial translocation are poorly defined . Possible routes for transmucosal passage of bacteria include transcellular and paracellular channels . Bacterial engulfment is a prerequisite for transcellular transport . To determine whether transcellular transport is required for transmucosal bacterial passage, we examined the effect of various inhibitors of endocytosis, such as colchicine, cytochalasin B, and sodium fluoride on transmucosal passage of bacteria across an ileal mucosal membrane mounted in the Ussing chamber . Colchicine and sodium fluoride increased the rate of decline of the potential difference across the membranes . However, neither colchicine, cytochalasin B, nor sodium fluoride affected the incidence of transmucosal bacterial passage . Sodium fluoride, which depletes intracellular ATP, significantly decreased the number of bacteria that passed per membrane . Our data suggest that transcellular transport may not be required for spontaneous transmucosal passage of bacteria, and furthermore bacterial passage may be, at least in part, an energy-dependent process. Proc Natl Acad Sci U S A, 2003 Mar 4, 100(5), 2695 - 9 Epub 2003 Feb 19. Bacterial enterotoxins are associated with resistance to colon cancer; Pitari GM et al.; One half million patients suffer from colorectal cancer in industrialized nations, yet this disease exhibits a low incidence in under-developed countries . This geographic imbalance suggests an environmental contribution to the resistance of endemic populations to intestinal neoplasia . A common epidemiological characteristic of these colon cancer-spared regions is the prevalence of enterotoxigenic bacteria associated with diarrheal disease . Here, a bacterial heat-stable enterotoxin was demonstrated to suppress colon cancer cell proliferation by a guanylyl cyclase C-mediated signaling cascade . The heat-stable enterotoxin suppressed proliferation by increasing intracellular cGMP, an effect mimicked by the cell-permeant analog 8-br-cGMP . The antiproliferative effects of the enterotoxin and 8-br-cGMP were reversed by L-cis-diltiazem, a cyclic nucleotide-gated channel inhibitor, as well as by removal of extracellular Ca(2+), or chelation of intracellular Ca(2+) . In fact, both the enterotoxin and 8-br-cGMP induced an L-cis-diltiazem-sensitive conductance, promoting Ca(2+) influx and inhibition of DNA synthesis in colon cancer cells . Induction of this previously unrecognized antiproliferative signaling pathway by bacterial enterotoxin could contribute to the resistance of endemic populations to intestinal neoplasia, and offers a paradigm for targeted prevention and therapy of primary and metastatic colorectal cancer. FEMS Microbiol Lett, 2003 Feb 14, 219(1), 121 - 7 A new method for rapid screening of bacterial species- or subspecies-specific DNA probes; Kook JK et al.; A simple assay for the rapid screening of bacterial species- or subspecies-specific DNA probes for the random cloning method is presented, involving the use of genomic DNAs as probes and recombinant plasmid DNAs containing genomic DNA digested with HindIII as targets . The optimal amount of target DNAs and the concentration of digoxigenin-labeled genomic DNA probes were 20 ng and 100 ng ml(-1) (or 10 ng and 200 ng ml(-1)), respectively . The method was applied to the development of Fusobacterium nucleatum subspecies-specific probes . Our results showed that four out of 96 probes were F . nucleatum subspecies-specific, which was confirmed by Southern blot analysis . Our results indicate that the new method can be used for the rapid screening of species- or subspecies-specific probes. Probl Tuberk, 2002, (10), 16 - 9 {Methods of the detection of non-bacterial tuberculosis and its clinical and X-ray characteristics in children and adolescents}; Gubkina MF; Abacillary tuberculosis is presented by all clinical forms among which tuberculosis of intrathoracic lymph nodes, infiltrative and focal pulmonary tuberculosis are common . Abacillary tuberculous processes are restrictive: the involvement of 1-2 segments of the lung is observed in 95% of cases, that of 1-2 groups of intrathoracic lymph nodes is in 87.5% . Detection rates are 36% at tuberculin diagnosis, 30% at fluorography, and 14% on examination of those in contact with a patient with tuberculosis . Abacillary tuberculosis is asymptomatic . Its major clinical (intoxication and pulmonary) syndromes are commonly mild in 97.8% . The clinical manifestations of the disease are somewhat more pronounced in patients with infiltrative pulmonary tuberculosis. Zhonghua Nan Ke Xue, 2002 Dec, 8(6), 442 - 4 {Influence of male genital bacterial infection on sperm function}; Li HY et al.; Male infertility is related to urogenital tract bacterial infection . The significance of bacteriosperimia for male infertility has been paid attention in recent years . Asymptomatic bacteriosperm may play an important role . However, litter is known about the adverse influence of some uropathogenic bacteria on sperm function . The article reviewed some uropathogenic bacteria in male urogential tract and several pathomechanisms of effect bacteria on sperm function. Zhongguo Zhong Xi Yi Jie He Za Zhi, 2002 Jul, 22(7), 505 - 7 {Clinical observation on dusuqing in treating senile bacterial pneumonia}; Li JS et al.; OBJECTIVE: To evaluate the clinical effect of Dusuqing (DSQ) in treating senile bacterial pneumonia and to explore its possible mechanism . METHODS: Patients were randomly divided into two groups, the treated group treated with conventional western therapy (CWT) and the combined treated group treated with CWT plus DSQ respectively . The changes of serum cytokines, interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-alpha), before and after treatment were observed . RESULTS: In the 33 patients of the combined treated group, the total effective rate being 93.9% . While in the 31 patients of treated group, the total effective rate being 83.9% . The difference of the total effective rate between the two groups was significant (P < 0.05) . The scores of principal symptoms, secondary symptoms and total scores were all obviously lowered in both groups, particularly in the combined treated group . The peripheral blood IL-2 level was lower and TNF-alpha higher in senile patients than those in healthy aged subjects respectively . All the two parameters approached normal level in the combined treated group but with insignificant change in the CWT treated group . CONCLUSION: DSQ showed obvious effect in treating senile bacterial pneumonia . The mechanism might be related with the lowering of inflammatory cytokines such as oversecreted TNF-alpha, as well as the improvement of immune function (elevation of IL-2) of organism. Am J Obstet Gynecol, 2003 Feb, 188(2), 318 - 23 Vaginal polymorphonuclear leukocytes and bacterial vaginosis as markers for histologic endometritis among women without symptoms of pelvic inflammatory disease; Yudin MH et al.; OBJECTIVE: The study was performed to determine whether vaginal polymorphonuclear leukocytes can be used as predictors of histologic endometritis among women at risk for, but without symptoms of, acute pelvic inflammatory disease . STUDY DESIGN: Five hundred thirty-seven women with, or at risk for, pelvic infection underwent pelvic examinations, including endometrial biopsies . These women were assessed for the presence of vaginal polymorphonuclear leukocytes, bacterial vaginosis, sexually transmitted diseases, and histologic endometritis . RESULTS: Vaginal neutrophils were present in 240 (44.7%) and histologic endometritis was present in 77 (14.3%) of the study population . Women with histologic endometritis were significantly more likely to have vaginal neutrophils present, with an odds ratio of 3.2 (95% CI 1.9-5.7) . When study subjects were stratified by the presence of sexually transmitted diseases and vaginal neutrophils, the prevalence of histologic endometritis was highest in women with both conditions present, with an odds ratio of 7.0 (95% CI 3.5-14.3) . When subjects were stratified by the presence of bacterial vaginosis and vaginal neutrophils, the prevalence of histologic endometritis was again highest in women with both conditions present, with an odds ratio of 4.8 (95% CI 1.4-16.3) . The presence of vaginal neutrophils diagnosed by saline wet mount had a high sensitivity (90.9%) and negative predictive value (94.5%), but a low specificity (26.3%) and positive predictive value (17.1%) for the diagnosis of upper genital tract infection . CONCLUSION: The presence of vaginal polymorphonuclear leukocytes has a high sensitivity and negative predictive value for the diagnosis of upper genital tract infection. Am J Gastroenterol, 2003 Feb, 98(2), 327 - 31 Is irritable bowel syndrome more common in patients presenting with bacterial gastroenteritis? A community-based, case-control study; Parry SD et al.; OBJECTIVE: Irritable bowel syndrome (IBS) has been reported to follow infectious diarrhea . Food-borne infections affect 76 million people in the United States and 9.4 million in England per year; of these, only a small percentage of patients see their doctor, and even fewer will have stool culture confirmation . We hypothesized that patients who present to their doctor with gastroenteritis and have positive stool samples may be different from the normal population with regard to their pre-existing bowel symptoms . Our aim was to determine if patients with bacterial gastroenteritis were more likely to have prior IBS, functional dyspepsia, or functional diarrhea, compared with a control population . METHODS: Between January, 2000 and January, 2001, subjects with stool positive bacterial gastroenteritis and control subjects from the same primary care practice were invited to participate . The main outcome measure was the presence of IBS, functional dyspepsia, or functional diarrhea diagnosed using self-report Rome II modular questionnaires . RESULTS: A total of 217 people with recent bacterial gastroenteritis and 265 community controls consented to participate in the study . Of these, 89/217 cases and 46/265 controls had one of the functional GI disorders (OR = 3.3; 95% CI = 2.17-5.00) . IBS was present in 67 cases (31%) and 26 controls (10%) (OR = 4.1; 95% CI = 2.49-6.72) . There was no statistically significant difference in the presence of prior functional dyspepsia or functional diarrhea . CONCLUSIONS: IBS is more frequent before diagnosis in people with bacterial gastroenteritis presenting to their primary care physician than in community controls . Studies that examine the rate of IBS after bacterial gastroenteritis need to carefully exclude people with prior IBS in a systematic way. Int J STD AIDS, 2003 Jan, 14(1), 6 - 10 Relation between Gram-stain and clinical criteria for diagnosing bacterial vaginosis with special reference to Gram grade II evaluation; Taylor-Robinson D et al.; The aim of this study was to analyse how the results of Gram-staining vaginal smears correlated with the clinical criteria for determining the existence of bacterial vaginosis (BV) and, in particular, how the category defined as 'intermediate' or Gram grade II did so and its significance . Women attending an antenatal clinic with an abnormal vaginal flora, that is those who had Gram-stains of grades II or III, the latter considered to equate with BV, were given clindamycin or a placebo intravaginally and examined again on up to three occasions . Gram-stain readings of grade III correlated with the clinical criteria for BV on 356 (91.7%) of 388 occasions . Grade II readings covered the spectrum of clinical criteria and correlated with those for BV on 35 (37.2%) of 94 occasions . Grade I, recorded 231 times and seen usually after clindamycin treatment, was associated with BV only once . The sensitivity, specificity, positive predictive value and negative predictive value of the Gram stain for the diagnosis of BV, based on a combination of grades II and III, were 99.7%, 71.6%, 81% and 99.6%, respectively; based on grade III only, the values were 99.7%, 87.7%, 91.6% and 99.6%, respectively . Women reported a malodorous vaginal discharge on 49.2% of the occasions a grade III flora was seen and 13.3% of the times grade II was recorded . It was not associated with grade I and would seem a useful adjunct to the accepted clinical criteria for diagnosing BV . Each of the clinical criteria was found in about equal proportions (87%-91%) for women whose Gram grade was III . For grade II, an increased discharge was noted most often (76.5%) and 'clue' cells least often (24.5%) . A positive amine test was the most specific, being associated with <1% of grade I smears . Of women with grade III status, 91% reverted to grade I after treatment with clindamycin for three days . In contrast, of women with grade II status, 53% reverted to grade I, as did 47% of those who were given a placebo . The 'intermediate' (grade II) category is a Gram-stain diagnosis and not one that can be made clinically . It is important to recognize as a distinct entity not only because amalgamation with grade III diminishes the specificity and positive predictive value of the Gram-stain for diagnosing BV, but also because women of grade II status usually fail to respond to clindamycin treatment, whereas those of grade III do not. BMC Genomics . 2003 Feb 3;4(1):5 {Epub ahead of print} Ancient conserved domains shared by animal soluble guanylyl cyclases and bacterial signaling proteins; Iyer LM et al.; BACKGROUND: Soluble guanylyl cyclases (SGCs) are dimeric enzymes that transduce signals downstream of nitric oxide (NO) in animals . They sense NO by means of a heme moiety that is bound to their N-terminal extensions . RESULTS: Using sequence profile searches we show that the N-terminal extensions of the SGCs contain two globular domains . The first of these, the HNOB (Heme NO Binding) domain, is a predominantly alpha-helical domain and binds heme via a covalent linkage to histidine . Versions lacking this conserved histidine and are likely to interact with heme non-covalently . We detected HNOB domains in several bacterial lineages, where they occur fused to methyl accepting domains of chemotaxis receptors or as standalone proteins . The standalone forms are encoded by predicted operons that also contain genes for two component signaling systems and GGDEF-type nucleotide cyclases . The second domain, the HNOB associated (HNOBA) domain occurs between the HNOB and the cyclase domains in the animal SGCs . The HNOBA domain is also detected in bacteria and is always encoded by a gene, which occurs in the neighborhood of a gene for a HNOB domain . CONCLUSION: The HNOB domain is predicted to function as a heme-dependent sensor for gaseous ligands, and transduce diverse downstream signals, in both bacteria and animals . The HNOBA domain functionally interacts with the HNOB domain, and possibly binds a ligand, either in cooperation, or independently of the latter domain . Phyletic profiles and phylogenetic analysis suggest that the HNOB and HNOBA domains were acquired by the animal lineage via lateral transfer from a bacterial source. Biochemistry, 2003 Feb 25, 42(7), 2016 - 24 B-side charge separation in bacterial photosynthetic reaction centers: nanosecond time scale electron transfer from HB- to QB; Kirmaier C et al.; We report time-resolved optical measurements of the primary electron transfer reactions in Rhodobacter capsulatus reaction centers (RCs) having four mutations: Phe(L181) --> Tyr, Tyr(M208) --> Phe, Leu(M212) --> His, and Trp(M250) --> Val (denoted YFHV) . Following direct excitation of the bacteriochlorophyll dimer (P) to its lowest excited singlet state P, electron transfer to the B-side bacteriopheophytin (H(B)) gives P(+)H(B)(-) in approximately 30% yield . When the secondary quinone (Q(B)) site is fully occupied, P(+)H(B)(-) decays with a time constant estimated to be in the range of 1.5-3 ns . In the presence of excess terbutryn, a competitive inhibitor of Q(B) binding, the observed lifetime of P(+)H(B)(-) is noticeably longer and is estimated to be in the range of 4-8 ns . On the basis of these values, the rate constant for P(+)H(B)(-) --> P(+)Q(B)(-) electron transfer is calculated to be between approximately (2 ns)(-)(1) and approximately (12 ns)(-)(1), making it at least an order of magnitude smaller than the rate constant of approximately (200 ps)(-)(1) for electron transfer between the corresponding A-side cofactors (P(+)H(A)(-) --> P(+)Q(A)(-)) . Structural and energetic factors associated with electron transfer to Q(B) compared to Q(A) are discussed . Comparison of the P(+)H(B)(-) lifetimes in the presence and absence of terbutryn indicates that the ultimate (i.e., quantum) yield of P(+)Q(B)(-) formation relative to P is 10-25% in the YFHV RC. Vet Ther, 2002 Winter, 3(4), 381 - 6 Bacterial cure and somatic cell count response of dairy cows with a positive California Mastitis Test at calving to therapy with cephapirin sodium; Rosenberg JB et al.; Cows without signs of clinical mastitis were evaluated by the California Mastitis Test at calving (Day 0) . Milk samples from 117 of 184 quarters (64 cows) were positive by this test for mastitis and were submitted for bacterial culture and determination of somatic cell counts . Cows with infected quarters were randomly allocated to treatment with cephapirin sodium by intramammary infusion or to be untreated as controls . Two and 4 weeks following calving, milk was again sampled from the infected quarters and tested . By the 4-week evaluation, the quarters treated with cephapirin sodium had significantly (P < or = .05) fewer positive bacterial cultures and somatic cell counts were significantly (P < or =.05) reduced compared with untreated control quarters. Curr Cardiol Rep, 2003 Mar, 5(2), 129 - 36 Embolic risk in subacute bacterial endocarditis: determinants and role of transesophageal echocardiography; Habib G; Embolic events are a frequent and life-threatening complication of subacute bacterial endocarditis (SBE), and are associated with a high mortality and morbidity . The prediction of the embolic risk in an individual patient remains a challenge . Echocardiography plays a key role in the management of infective endocarditis both for the diagnostic and prognostic assessment of these patients, but its value in predicting embolic events is still debated . This review focuses on the characteristics of embolic events in SBE, and on the role of echocardiography, especially transesophageal echocardiography (TEE), in the prediction of embolic events . The review confirms that, among other factors, TEE may be helpful in both the prediction of the embolic risk, and the decision to perform early surgery. Org Lett, 2003 Feb 20, 5(4), 539 - 41 Synthesis of a carbohydrate-derived hydroxamic acid inhibitor of the bacterial enzyme (LpxC) involved in lipid A biosynthesis; Li X et al.; {reaction: see text} The enzyme LpxC (UDP-3-O-{(R)-3-hydroxymyristoyl}-GlcNAc deacetylase) catalyzes the second step of lipid A biosynthesis and is essential for bacterial growth . A GlcNAc-derived hydroxamic acid inhibitor 8 of this enzyme was synthesized using two different routes . Compound 8 exhibits activity toward LpxC enzymes from a wider spectrum of bacterial species than any of the previously reported hydroxamic acid inhibitors. Rev Esp Quimioter, 2002 Mar, 15(1), 25 - 31 {Mechanisms of bacterial resistance to quinolones}; Talens-Visconti R et al.; In order to produce their cytotoxic effect, quinolones must enter the cell through the bacterial membrane to reach their target, DNA-gyrase or topoisomerase IV, and induce cell death . The mechanisms of resistance to fluoroquinolones include: those mediated by gene mutations codifying for DNA gyrase and topoisomerase IV and leading to QRDR; those characterized by changes in the permeability of the outer membrane which decrease intracellular penetration of the drug; and those caused by active endogenous carriers responsible for drug efflux . These resistance mechanisms can occur alone or in combination; in fact, it is believed that high resistance levels to quinolones in vivo are associated with simultaneous mechanisms . This article reviews such resistance mechanisms and establishes, when possible, their relation to the structure of quinolones. Curr Opin Struct Biol, 2003 Feb, 13(1), 31 - 9 Bacterial RNA polymerases: the wholo story; Murakami KS et al.; Recent structural and biophysical results have provided unprecedented insights into the structure and function of the bacterial RNA polymerase holoenzyme as it goes through the steps of transcription initiation . Comparisons with structural analyses of evolutionarily unrelated RNA polymerases reveal unexpected general features of the initiation process. Yan Ke Xue Bao, 2000 Jun, 16(2), 146 - 8 {Clinical investigation of 0.3% levofloxacin eyedrops on the treatment of cases with acute bacterial conjunctivitis and bacterial keratitis}; Zhang M et al.; PURPOSE: To investigate the effects of 0.3% levofloxacin eyedrops on the treatment of acute bacterial conjunctivitis and bacterial keratitis . METHODS: The patients were randomly divided into treated group and control group . They were given 0.3% levofloxacin and 0.3% ofloxacin eyedrops respectively . RESULTS: From April to December in 1999, 132 cases were enrolled in the study . It showed that there was no significant difference between two drops in the cure and effective rates, but the mean cure day of keratitis in levofloxacin group was significantly shorter than that in ofloxacin group . No serious side effects occurred . CONCLUSION: Levofloxacin eyedrops was effective and safe antibiotics in treating bacterial conjunctivitis and keratitis. Extremophiles, 2003 Feb, 7(1), 9 - 16 Epub 2002 Sep 07. Citrate synthase from Thermus aquaticus: a thermostable bacterial enzyme with a five-membered inter-subunit ionic network; Nordberg Karlsson E et al.; A bacterial thermostable citrate synthase has been analyzed to investigate the structural basis of its thermostability, and to compare such features with those previously identified in archaeal citrate synthases . The gene encoding the citrate synthase from Thermus aquaticus was identified from a gene library by screening with a PCR fragment amplified from genomic DNA using a primer based on the determined N-terminal amino acid sequence and a citrate synthase consensus primer . Apart from high sequence similarities with citrate synthase sequences within the Thermus/ Deinococcus group, the analyzed enzyme has highest similarities with the enzyme from the hyperthermophilic Archaeon Pyrococcus furiosus . The recombinant enzyme is a dimer with high specific activity . Compared to its thermoactivity (T(opt)at 80 degrees C), the thermal stability of the enzyme is high, as judged from its T(m) (101 degrees C), and from irreversible thermal inactivation assays . Molecular modeling of the structure revealed an inter-subunit ion-pair network, comparable in size to the network found in the citrate synthase from P . furiosus; these networks are discussed in relation to the high thermal stability of these bacterial and archaeal enzymes. Biochemistry, 2003 Feb 18, 42(6), 1718 - 30 Quinone reduction via secondary B-branch electron transfer in mutant bacterial reaction centers; Laible PD et al.; Symmetry-related branches of electron-transfer cofactors-initiating with a primary electron donor (P) and terminating in quinone acceptors (Q)-are common features of photosynthetic reaction centers (RC) . Experimental observations show activity of only one of them-the A branch-in wild-type bacterial RCs . In a mutant RC, we now demonstrate that electron transfer can occur along the entire, normally inactive B-branch pathway to reduce the terminal acceptor Q(B) on the time scale of nanoseconds . The transmembrane charge-separated state P(+)Q(B)(-) is created in this manner in a Rhodobacter capsulatus RC containing the F(L181)Y-Y(M208)F-L(M212)H-W(M250)V mutations (YFHV) . The W(M250)V mutation quantitatively blocks binding of Q(A), thereby eliminating Q(B) reduction via the normal A-branch pathway . Full occupancy of the Q(B) site by the native UQ(10) is ensured (without the necessity of reconstitution by exogenous quinone) by purification of RCs with the mild detergent, Deriphat 160-C . The lifetime of P(+)Q(B)(-) in the YFHV mutant RC is >6 s (at pH 8.0, 298 K) . This charge-separated state is not formed upon addition of competitive inhibitors of Q(B) binding (terbutryn or stigmatellin) . Furthermore, this lifetime is much longer than the value of approximately 1-1.5 s found when P(+)Q(B)(-) is produced in the wild-type RC by A-side activity alone . Collectively, these results demonstrate that P(+)Q(B)(-) is formed solely by activity of the B-branch carriers in the YFHV RC . In comparison, P(+)Q(B)(-) can form by either the A or B branches in the YFH RC, as indicated by the biexponential lifetimes of approximately 1 and approximately 6-10 s . These findings suggest that P(+)Q(B)(-) states formed via the two branches are distinct and that P(+)Q(B)(-) formed by the B side does not decay via the normal (indirect) pathway that utilizes the A-side cofactors when present . These differences may report on structural and energetic factors that further distinguish the functional asymmetry of the two cofactor branches. Shock, 2003 Feb, 19(2), 144 - 9 Natural killer cells participate in bacterial clearance during septic peritonitis through interactions with macrophages; Godshall CJ et al.; Natural killer (NK) cells have a well-established role in host defense against viral infections and malignancies . However, their function in bacterial infection and sepsis is poorly defined . We hypothesized that NK cells, as a major producer of interferon-gamma during sepsis, would be important in host defense against bacterial infections . Cecal ligation and puncture (CLP) was performed on Swiss Webster mice depleted of NK cells by pretreatment with anti-asialo GM1 and control mice given immunoglobulin G (IgG) antibody . NK cell-depleted mice had significantly higher anaerobic bacterial counts in the liver and peritoneal lavage fluid, as well as higher aerobic counts in the liver and blood 4 h after CLP . Macrophage phagocytosis, nitric oxide production, and interleukin (IL)-6 levels at 4 h were also decreased in mice depleted of NK cells compared with controls . Greater neutrophil influx into the peritoneum, indicated by higher myeloperoxidase levels, was also seen in NK cell-depleted mice . At 8 and 18 h after CLP, bacterial counts were similar between groups, and overall survival rates were not significantly different . Peritoneal IL-12 levels significantly increased by 18 h in normal mice, but not in NK cell-depleted animals . Our data suggest that NK cells participate in the early local and systemic eradication of bacteria and regulation of IL-12 during polymicrobial sepsis . These effects are likely due to their interactions with macrophages. Anal Biochem, 2003 Feb 1, 313(1), 76 - 85 Time-minimized determination of ribosome and tRNA levels in bacterial cells using flow field-flow fractionation; Arfvidsson C et al.; The evaluation of the translation capacity of cells that produce recombinant proteins can be made by monitoring their ribosomal composition . In a previous use of asymmetrical flow field-flow fractionation (AsFlFFF) for this purpose the overall analysis time was more than 1 h and 40 min, based on a standard protocol for cell harvest, washing, cell disruption, and the final 8-min AsFlFFF determination of ribosome and subunits . In the present work the overall analysis time was reduced to 16 min . The washing step was deleted and a time-consuming freeze-thaw cycle . Cell disruption was obtained by a time-minimized lysozyme and detergent treatment for 1.5 min, respectively . The ribosomal material was finally fractionated and quantified in only 6 min, without previous centrifugation, using AsFlFFF . The great time reduction will enable the future use of AsFlFFF at-line to a growing cell cultivation, continuously monitoring the change in ribosomal composition or in other applications requiring high sample throughput . To demonstrate the high efficiency of the method the ribosome and tRNA composition in an Escherichia coli cultivation was monitored every half an hour, giving 18 measurements across the complete growth curve, a frequency of data enough to make decisions about induction or termination of the cultivation . Hua Xi Yi Ke Da Xue Xue Bao, 2002 Apr, 33(2), 291 - 3, 325 {Multicenter evaluation on the efficacy and safety of sparfloxacin in the treatment of acute bacterial infections}; Cai Y et al.; OBJECTIVE: To evaluate the efficacy and safety of domestic sparfloxacin . METHODS: A multicentre randomized controlled clinical trial was conducted to compare sparfloxacin versus ofloxacin in the treatment of acute bacterial infections . 58 patients received 200 mg of sparfloxacin once daily intravenous drip and 61 patients received 200 mg of ofloxacin twice daily intravenous drip . Both durgs were given for 7-14 days . RESULTS: The clinical cure rates of sparfloxacin group and ofloxacin group were 44.83% and 42.62%, and the efficacy rates were 87.93% and 81.97%, respectively; no significant differences were noted between the two groups (P > 0.05) . The bacterial eradication rates were 86.00% and 84.62% respectively (P > 0.05) . The adverse reactions in the two groups were 13.79% and 22.58% (P > 0.05) . Photosensitivity reaction was not observed in this study . CONCLUSION: Domestic sparfloxacin is effective and safe in treating acute bacterial infections. Proc Natl Acad Sci U S A, 2003 Feb 18, 100(4), 1574 - 9 Epub 2003 Feb 06. Assembly of core helices and rapid tertiary folding of a small bacterial group I ribozyme; Rangan P et al.; Compact but non-native intermediates have been implicated in the hierarchical folding of several large RNAs, but there is little information on their structure . In this article, ribonuclease and hydroxyl radical cleavage protection assays showed that base pairing of core helices stabilize a compact state of a small group I ribozyme from Azoarcus pre-tRNA(ile) . Base pairing of the ribozyme core requires 10-fold less Mg(2+) than stable tertiary interactions, indicating that assembly of helices in the catalytic core represents a distinct phase that precedes the formation of native tertiary structure . Tertiary folding occurs in <100 ms at 37 degrees C . Such rapid folding is unprecedented among group I ribozymes and illustrates the association between structural complexity and folding time . A 3D model of the Azoarcus ribozyme was constructed by identifying homologous sequence motifs in rRNA . The model reveals distinct structural features, such as a large interface between the P4-P6 and P3-P9 domains, that may explain the unusual stability of the Azoarcus ribozyme and the cooperativity of folding. Curr Pharm Des, 2003, 9(1), 11 - 24 Mast cell cytokine and chemokine responses to bacterial and viral infection; Marshall JS et al.; Mast cells have been most widely studied in the context of allergic disease but also play a critical role in host defence against bacterial infection, most elegantly demonstrated in studies using mast cell deficient w/wv mice . There is less data available concerning the role of mast cells in defence against viral pathogens, however, mast cells have been demonstrated to be a potential reservoir of infection for several pathogens, such as HIV-1 and dengue, and capable of producing mediators following challenge with a number of viral products . Traditional mast cell mediators such as histamine, protease enzymes and leukotrienes are important for effective host responses . The cytokines and chemokines produced by mast cells in response to pathogens are known to profoundly alter the nature of the innate immune response and its effectiveness in eliminating infection . Cytokine and chemokine production by mast cells is closely regulated and may occur independently of classical mast cell degranulation . Depending upon the nature of the stimulus or type of infection, a unique profile of cytokines is induced . In this review, we will examine the role and regulation of mast cell cytokines and chemokines in the context of a number of bacterial and viral infections, emphasizing the multiple receptor mechanisms used to activate mast cells . This area of research is still in its early stages and much work remains to be done . However, understanding the unique properties of resident tissue mast cells and how their cytokine responses are regulated by pathogens or pathogen products, will provide important opportunities for the therapeutic manipulation of local immune responses. Trends Microbiol, 2002 Dec, 10(12), 547 - 50 Molecular basis of bacterial virulence and survival within infected hosts and the environment; Raupach B et al.; The EMBO-FEBS lecture course on the 'Molecular Basis of Bacterial Virulence and Survival Within Infected Hosts and in the Environment', organized by Pascale Cossart, Efstathios Gonos and Roberto Kolter, was held on the island of Spetsai, Greece, September 3-13, 2002. Wei Sheng Yan Jiu, 2001 Sep, 30(5), 315 - 6, 318 {Study on the elimination of bacterial endotoxins by ultrafiltration device}; Wang Z et al.; An external-pressed polysulfone hollow fiber UF membrane, was made by phase inversion method . The cutoff molecular weight was 6000 . The water permeation rate of this membrane module (psi 90 x 500 mm) is more than 150 L/hr.atm . The operating results show that the elimination of bacterial endotoxins by this membrane has reached the national standard in pharmacopoeia, and this technology is worthy to be extended over to medical industry . In addition, the essence of pyreotogen, the quantification units and measure methods for pyreotogen, and the operating conditions of UF device in the process of eliminating pyreotogen are introduced . Some critical points in operation was discussed. Chemistry, 2002 Oct 18, 8(20), 4598 - 612 Second-generation mimics of ganglioside GM1 oligosaccharide: a three-dimensional view of their interactions with bacterial enterotoxins by NMR and computational methods; Bernardi A et al.; As a step to delineate a strategy of ligand design for cholera toxin (CT), NMR studies were performed on several mimics of the GM1 ganglioside oligosaccharide . The conformation of these analogues was investigated first in solution and then upon binding to cholera toxin by transferred nuclear Overhauser effect (TR-NOE) measurements . It was demonstrated that CT selects a conformation similar to the global minima of the free saccharides from the ensemble of presented conformations . No evidence of major conformational distortions was obtained, but one or two of the available conformers of the hydroxyacid side chain appear to be selected in the bound state . The NMR data were interpreted with the aid of computer models, generated and analyzed by using a combination of different approaches (MacroModels' MC/EM and MC/SD, Autodock, and GRID) . Analysis of the NMR data supported by computational studies allowed us to interpret the experimental observations and to derive workable models of the ligand:toxin complexes . These models suggest that the higher affinity of the (R)-lactic acid derivative 3 may stem from lipophilic interactions with a hydrophobic area in the toxin binding site located in the vicinity of the sialic acid side chain binding region of the CT:GM1 complex, and formed by the side chain of Ile-58 and Lys-34 . Thus, the models obtained have allowed us to make useful design suggestions for the improvement of ligand affinity. Eur J Gastroenterol Hepatol, 2003 Feb, 15(2), 145 - 50 Intestinal mucosal oxidative damage and bacterial translocation in cirrhotic rats; Chiva M et al.; BACKGROUND: Bacterial translocation plays an important role in the pathogenesis of spontaneous bacterial peritonitis mainly due to intestinal bacterial overgrowth . Alterations in the functional integrity of the intestinal barrier caused by an increased production of free radical metabolites as a consequence of portal hypertension could also facilitate bacterial translocation in cirrhotic rats . OBJECTIVE: The aim of the study was to determine intestinal mucosal lipid peroxidation and neutrophil infiltration and their relationship with portal hypertension and bacterial translocation in cirrhotic rats . DESIGN: Eighteen male Sprague-Dawley rats with cirrhosis induced by carbon tetrachloride, administered by gavage, and eight control rats were included in the study . METHODS: Samples of jejunum, ileum and caecum were obtained by laparotomy for the determination of malondialdehyde and myeloperoxidase as indexes of lipid peroxidation and neutrophil infiltration, respectively . Samples of ascitic and pleural fluids, mesenteric lymph nodes and ileal stools were obtained for the culture of microoganisms . RESULTS: The concentration of malondialdehyde was significantly higher in ileal and caecal, but not in jejunal mucosa, in cirrhotic rats, mainly in those with ascites (P< 0.01), as compared to control rats (P< 0.01), and in cirrhotic rats with bacterial translocation compared to those without bacterial translocation (P< 0.01) . No differences between groups were observed in the concentrations of myeloperoxidase in jejunum, ileum or caecum . A direct correlation between ileal malondialdehyde and portal pressure was observed (P< 0.01) . CONCLUSIONS: Cirrhotic rats, particularly those with ascites and bacterial translocation, show increased malondialdehyde levels in ileal and caecal mucosa . These results suggest that mucosal oxidative damage in ileum and caecum could favour bacterial translocation in cirrhotic rats. J Gen Virol, 2003 Feb, 84(Pt 2), 301 - 6 Mutagenesis of a bovine herpesvirus type 1 genome cloned as an infectious bacterial artificial chromosome: analysis of glycoprotein E and G double deletion mutants; Trapp S et al.; The genome of bovine herpesvirus type 1 Schonboken was cloned as a bacterial artificial chromosome (BAC) by inserting mini F plasmid sequences into the glycoprotein (g) E gene . The resulting BAC clone, pBHV-1DeltagE, was transfected into bovine kidney cells and viable gE-negative BHV-1 (BHV-1DeltagE) was recovered . By RecE/T mutagenesis in Escherichia coli, the gG open reading frame was deleted from pBHV-1DeltagE . From the mutated BAC, double negative BHV-1DeltagE-gG was reconstituted and its growth properties were compared to those of rescuant viruses in which the gE gene was restored (BHV-1rev, BHV-1DeltagG) . The mutant viruses did not exhibit markedly lowered virus titres . Plaque sizes of BHV-1DeltagE, BHV-1DeltagE-gG and BHV-1DeltagG, however, were reduced by 19 to 55 % compared to parental strain Schonboken or BHV-1rev . Our results suggested that gE and gG function independently from each other in cell-to-cell spread, because an additive effect on plaque formation was observed in the gE/gG double deletion mutant. Lancet, 2003 Jan 25, 361(9354), 275 - 80 Recombinant human interleukin 11 and bacterial infection in patients with {correction of} haematological malignant disease undergoing chemotherapy: a double-blind placebo-controlled randomised trial; Ellis M et al.; BACKGROUND: Bacteraemia in patients with haematological malignant disease causes substantial morbidity . Recombinant human interleukin 11 (rhIL-11) prevents gastrointestinal epithelial disintegrity and has immunomodulatory actions . Our aim was to ascertain whether or not treatment with rhIL-11 can prevent gut-associated infections . METHODS: We did a double-blind placebo-controlled randomised trial, to which we enrolled 40 patients with haematological malignant disease who were undergoing chemotherapy . Patients received either rhIL-11 50 microg/kg (n=20) or placebo (n=20) daily by subcutaneous injection from the day before the start of chemotherapy until resolution of neutropenia or for 21 days, whichever was longer . Our primary outcome measure was a reduction in bacteraemia . Analysis was by intention to treat . FINDINGS: Significantly fewer patients who received rhIL-11 rather than placebo developed bacteraemia, particularly of gastrointestinal origin: the proportion of patients with at least one positive blood culture was 0.65 and 0.25, respectively (p=0.02) . The numbers of patients (placebo vs rhIL-11) for each number of distinct isolates were: no organism isolated seven versus 15, one organism nine versus four, two organisms two versus one, three organisms one versus none, and four organisms one versus none (p=0.01), suggesting a lower bacterial load in the rhIL-11 than in the placebo group . Time to first bacteraemic event was longer in patients who received rhIL-11 (p=0.03) than in those who received placebo . INTERPRETATION: rhIL-11 reduces the frequency and load of bacteraemia in patients with haematological malignant disease undergoing chemotherapy, possibly by gastrointestinal cytoprotective or immunological mechanisms {corrected}. Shock, 2003 Jan, 19(1), 5 - 12 Expression patterns of the lipopolysaccharide receptor CD14, and the FCgamma receptors CD16 and CD64 on polymorphonuclear neutrophils: data from patients with severe bacterial infections and lipopolysaccharide-exposed cells; Wagner C et al.; In polymorphonuclear neutrophils (PMN) CD14, one of the receptors for lipopolysaccharides (LPS) is stored intracellularly as a preformed protein, with only few receptors expressed on the surface . We now report that in patients with severe bacterial infections, CD14 expression is profoundly upregulated, as is CD64 (FcgammaRI), the high-affinity receptor for IgG, whereas CD16 (FcgammaRIII) was partly lost from the surface . To further analyze regulation of these receptors, PMN of healthy donors were exposed to low doses of LPS . By brief exposure (10-120 min) to LPS, CD14 was transferred to the surface in a cytochalasin B-sensitive manner, as were CD16 and CD64 . Prolonged culture (up to 48 h) resulted in a further upregulation of CD14, sustained expression of CD64, and profound decline of CD16, yielding a similar pattern of receptor expression as seen in the patients . Subsequent studies revealed that LPS induced de novo synthesis of CD14: the increase of surface expression could be inhibited by cycloheximide and by interfering with a known LPS-induced signaling event, the translocation of NFkappaB . Moreover, an up to 10-fold increase of specific mRNA was seen, as was incorporation into CD14 of 35S-methionine . The de novo synthesis prolonged expression of CD14, whereas the CD16 expression declined, generating a PMN phenotype characteristic for severe infection and indicative of escape from apoptosis of a PMN subpopulation. Mikrobiol Z, 2002 Sep-Oct, 64(5), 53 - 8 {Phagocytic activity of mouse peritoneal macrophages under administration of bacterial vaccines and prospidin}; Pavlenko NV; Phagocytic activity of peritoneal macrophages in cases of separate and combined injections of killed bacterial vaccines and prospidin has been studied in the work . As a result of researches conducted, it has been found out that the injection of vaccines together with prospidin makes it possible to reduce an inhibiting effect of prospidin on phagocytic activity of monocyte-macrophagal cells; first of all it concerns their digestive activity . Thus, the immunodepressing effect of antiblastome drugs may be reduced with the help of injection of killed bacterial vaccines which stimulate nonspecific factors of organism protection, including antineoplastic ones. Am J Hematol, 2003 Feb, 72(2), 115 - 20 Thromboxane synthesis is increased by upregulation of cytosolic phospholipase A2 and cyclooxygenase-2 in peripheral polymorphonuclear leukocytes during bacterial infection in childhood; Zaitsu M et al.; Prostaglandins (PGs) and thromboxane (TX) are important mediators of inflammation . Recent studies revealed that PG and TX synthesis is controlled by the regulation of PG- and TX-synthesizing enzymes . In this study, we examined the TX synthesis and the expression of TX-synthesizing enzymes in activated peripheral polymorphonuclear leukocytes (PMNs) obtained from children with bacterial infection . Blood samples were obtained from controls and patients with bacterial infection . A23187-stimulated production of TXB(2), a stable metabolite of TXA(2) in PMNs, was measured by a specific radioimmunoassay . The mRNA expression of cytosolic phospholipase A(2) (cPLA(2)), cyclooxygenase (COX)-1, COX-2, and TXA(2) synthase was determined by RT-PCR . The synthesis of TXB(2) in PMNs was significantly increased in the patients {925.0 (550.0-1100.0) pg/10(6) cells}, compared with the controls {550.0 (450.0-775.0) pg/10(6) cells} . The mRNA expression for cPLA(2) and COX-2 in PMNs was also enhanced in the patients . The results indicate that TX production in PMNs is significantly increased through possible transcriptional mechanisms of cPLA(2) and COX-2 during bacterial infection in children . The upregulation of TXA(2) synthesis may contribute to the process of acute inflammatory reaction caused by bacterial infection . Acta Orthop Scand, 2002 Dec, 73(6), 678 - 83 Bacterial contamination in postmortem bone donors; Vehmeyer S et al.; We analyzed factors influencing the contamination rate of allografts and blood samples obtained from postmortem bone donors . 5,710 allografts were harvested, from 550 donors of which 3,164 (55%) were swab culture negative . Blood cultures were positive in 140 donors (26%) . The risk of graft contamination increased with each extra team member (Odds Ratio 1.9) . It was also higher with organisms of greater virulence in donors with a positive blood culture (OR 3.5) . The risk of blood contamination increased per hour postmortem (OR 1.1) and the same increase was seen with organisms of high virulence . In donors with multiple trauma, the risk of blood contamination with organisms of high virulence was greater (OR 8.2), but smaller in donors with preceding organ procurement (OR 0.1) . To minimize the bacterial load, donors should be obtained in operating rooms, using aseptic techniques with only a few personnel for procurement . The postmortem time should be kept to a minimum . The procurement cultures from donors with multiple trauma should be carefully interpreted . Blood cultures should be taken into account, since these can help to find contamination not detected by swab cultures. J Cataract Refract Surg, 2003 Jan, 29(1), 20 - 6 Bacterial endophthalmitis after small-incision cataract surgery . effect of incision placement and intraocular lens type; Nagaki Y et al.; PURPOSE: To determine whether endophthalmitis after small-incision cataract surgery is affected by the incision site (superior sclerocorneal versus temporal cornea) or the foldable intraocular lens (IOL) material (silicone versus acrylic) . SETTING: Multicenter study . METHODS: Patients who had small-incision cataract surgery at Toyama Medical and Pharmaceutical University Hospital and affiliated hospitals from March 1998 to March 2001 were examined prospectively . The patients were randomized into 3 groups . In Group A, an acrylic IOL (MA60BM, Alcon) was implanted through a temporal corneal incision . In Group B, an acrylic IOL (MA60BM) was implanted via a superior sclerocorneal incision . In Group C, a silicone poly(methyl methacrylate) IOL (SI-40NB, Allergan) was implanted via a superior sclerocorneal incision . Each patient was followed for more than 6 months . RESULTS: Initially, 7622 patients (12 317 eyes) agreed to participate in the study . The final study included 3831 eyes in Group A, 3901 eyes in Group B, and 3863 eyes in Group C . Postoperative endophthalmitis was diagnosed clinically in 11 eyes (0.29%) in Group A, 2 (0.05%) in Group B, and 2 (0.05%) in Group C . Endophthalmitis proven by culture was found in 9 eyes in Group A, 2 in Group B, and 2 in Group C . The incidence of endophthalmitis in Group A was higher than in the other groups . The incidence of endophthalmitis in Group B was similar to that in Group C . The relative risk of postoperative endophthalmitis proven by culture in Groups B and C was 4.6 times (P =.037) lower than in Group A . CONCLUSION: The findings suggest that a temporal corneal incision may lead to an increased risk of postoperative endophthalmitis and that the IOL material does not affect the incidence of this complication. Biochemistry, 2003 Feb 4, 42(4), 1062 - 70 Genotoxicity and mutagenicity of chromium(VI)/ascorbate-generated DNA adducts in human and bacterial cells; Quievryn G et al.; Reduction of carcinogenic Cr(VI) by vitamin C generates ascorbate-Cr(III)-DNA cross-links, binary Cr(III)-DNA adducts, and can potentially cause oxidative DNA damage by intermediate reaction products . Here, we examined the mutational spectrum and the importance of different forms of DNA damage in genotoxicity and mutagenicity of Cr(VI) activated by physiological concentrations of ascorbate . Reduction of Cr(VI) led to a dose-dependent formation of both mutagenic and replication-blocking DNA lesions as detected by propagation of the pSP189 plasmids in human fibroblasts . Disruption of Cr-DNA binding abolished mutagenic responses and normalized the yield of replicated plasmids, indicating that Cr-DNA adducts were responsible for both mutagenicity and genotoxicity of Cr(VI) . The absence of DNA breaks and abasic sites confirmed the lack of a significant production of hydroxyl radicals and Cr(V)-peroxo complexes in Cr(VI)-ascorbate reactions . Ascorbate-Cr(III)-DNA cross-links were much more mutagenic than smaller Cr(III)-DNA adducts and accounted for more than 90% of Cr(VI) mutagenicity . Ternary adducts were also several times more potent in the inhibition of replication than binary complexes . The Cr(VI)-induced mutational spectrum consisted of an approximately equal number of deletions and G/C-targeted point mutations (51% G/C --> T/A and 30% G/C --> A/T) . In Escherichia coli cells, Cr(VI)-induced DNA adducts were only highly genotoxic but not mutagenic under either normal or SOS-induced conditions . Lower toxicity and high mutagenicity of ascorbate-Cr(III)-DNA adducts in human cells may result from the recruitment of an error-prone bypass DNA polymerase(s) to the stalled replication forks . Our results suggest that phosphotriester-type DNA adducts could play a more important role in human than bacterial mutagenesis. Lepr Rev, 2002 Dec, 73(4), 386 - 8 Relapse of lepromatous leprosy after WHO/MDT with rapid bacterial growth; Oliveira ML et al.; The authors report a case of relapse in a lepromatous patient 6 years after he had been cured by MDT/WHO/24 doses . The atypical aspect emphasized in this case is the bacterial load increase in a short period of time of 1 year after the smear count was negative, and the case reinforces the importance of patient education on release . No leprosy cases were identified in the patient's close contacts . It seems that relapse was a result of bacillary persistence, since a significant improvement was noted in relapsed lesions after two doses of MDT/WHO. Mol Biol Rep, 2002 Dec, 29(4), 363 - 8 A novel factor associating with the upstream regulatory element of murine Hoxa-7 induces bacterial cell death; Park S et al.; In order to understand the function of a cDNA (c171) associated with the upstream regulatory region of the Hoxa-7, the cDNA was cloned into the pGEX-4T-1 vector to produce it as a GST fusion protein . The size of the fusion protein was determined to be 48-kilodalton (kDa) . Sequence analysis revealed that a protein C171 contained one hydrophobic transmembrane domain in the N-terminal region and several putative phosphorylation and glycosylation sites . C171 protein inhibited the bacterial growth within 30 min after induction . The transmission electron microscopic examination revealed that the morphology of the cells expressing C171 was changed dramatically: i.e., unusually elongated phenotype compared with those of controls, and finally leading to a cell death . These results altogether indicate that a trace amount of C171 induces bacterial cell death. J Pediatr Gastroenterol Nutr, 2003 Feb, 36(2), 217 - 22 Endoscopic nodular gastritis: an endoscopic indicator of high-grade bacterial colonization and severe gastritis in children with Helicobacter pylori; Bahu Mda G et al.; OBJECTIVE: To investigate the significance of endoscopic nodular gastritis associated with Helicobacter pylori infection . METHODS: This prospective study included 185 children (50.8% boys) aged 1 to 12 years (mean, 6.9 +/- 3.0 years) who underwent upper intestinal endoscopy during evaluation of chronic abdominal pain . The authors assessed the endoscopic appearance of the stomach, noting those patients with endoscopic nodular gastritis . Urease activity of gastric mucosal biopsies was measured . With histologic examination, the presence and density of H . pylori organisms, the presence of follicular gastritis, the nature of inflammation, and the gastritis activity grade and overall gastritis score were assessed . RESULTS: H . pylori infection was identified in 50 children (27%) . Endoscopic nodular gastritis was significantly associated with active chronic gastritis and follicular gastritis . Nodularity in the stomach showed a high specificity (98.5%) and positive predictive value (91.7%) for the diagnosis of H . pylori infection and was observed in 22 of 50 (44%) H . pylori-positive patients and in 2 of 135 (1.5%) H . pylori-negative patients . A significant association was observed between older age and the prevalence of this finding (P< 0.001) . There was a significant increase in endoscopic nodular gastritis with increased H . pylori density and a positive correlation (Pearson coefficient = 0.97) with increased gastritis score on histologic examination . Increase in gastritis score was dependent on increased H . pylori density in patients with gastric nodularity; this finding was independent of age . CONCLUSIONS: Endoscopic findings of antral nodularity in children suggest the presence of H . pylori infection and follicular gastritis and may identify cases of severe gastritis and marked bacterial colonization. Biophys J, 2003 Feb, 84(2 Pt 1), 1146 - 60 Self-regulation phenomena applied to bacterial reaction centers: 2 . Nonequilibrium adiabatic potential: dark and light conformations revisited; Goushcha AO et al.; Experimental and theoretical results in support of nonlinear dynamic behavior of photosynthetic reaction centers under light-activated conditions are presented . Different conditions of light adaptation allow for preparation of reaction centers in either of two different conformational states . These states were detected both by short actinic flashes and by the switching of the actinic illumination level between different stationary state values . In the second method, the equilibration kinetics of reaction centers isolated from Rhodobacter sphaeroides were shown to be inherently biphasic . The fast and slow equilibration kinetics are shown to correspond to electron transfer (charge separation) at a fixed structure and to combined electron-conformational transitions governed by the bounded diffusion along the potential surface, respectively . The primary donor recovery kinetics after an actinic flash revealed a pronounced dependence on the time interval (deltat) between cessation of a lengthy preillumination of a sample and the actinic flash . A pronounced slow relaxation component with a decay half time of more than 50 s was measured for deltat > 10 s . This component corresponds to charge recombination in reaction centers for which light-induced structural changes have not relaxed completely before the flash . The amplitude of this component depended on the conditions of the sample preparation, specifically on the type of detergent used in the preparation . The redox potential parameters as well as the structural diffusion constants were estimated for samples prepared in different ways. DNA Repair (Amst), 2003 Mar 1, 2(3), 347 - 58 Budding yeast mms4 is epistatic with rad52 and the function of Mms4 can be replaced by a bacterial Holliday junction resolvase; Odagiri N et al.; MMS4 of Saccharomyces cerevisiae was originally identified as the gene responsible for one of the collection of methyl methanesulfonate (MMS)-sensitive mutants, mms4 . Recently it was identified as a synthetic lethal gene with an SGS1 mutation . Epistatic analyses revealed that MMS4 is involved in a pathway leading to homologous recombination requiring Rad52 or in the recombination itself, in which SGS1 is also involved . MMS sensitivity of mms4 but not sgs1, was suppressed by introducing a bacterial Holliday junction (HJ) resolvase, RusA . The frequencies of spontaneously occurring unequal sister chromatid recombination (SCR) and loss of marker in the rDNA in haploid mms4 cells and interchromosomal recombination between heteroalleles in diploid mms4 cells were essentially the same as those of wild-type cells . Although UV- and MMS-induced interchromosomal recombination was defective in sgs1 diploid cells, hyper-induction of interchromosomal recombination was observed in diploid mms4 cells, indicating that the function of Mms4 is dispensable for this type of recombination. Hua Xi Yi Ke Da Xue Xue Bao, 2000 Sep, 31(3), 402 - 4 {Value of intestinal decontamination by traditional Chinese medicine-X in the prevention of bacterial translocation complicated by severe acute pancreatitis in rats}; Qiao A et al.; This study was designed to evaluate the efficacy of traditional Chinese Medicine-X in preventing the necrotic infection of the pancreas in severe acute pancreatitis (SAP) . Sixty rats were randomly divided into five groups with 12 rats in each one: (1) normal control, (2) SAP + 0.9% normal saline (1 ml x 100 g-1 x 24 h-1), (3) SAP + gentamycin (2000 u x 100 g-1 x 24 h-1), (4) SAP + TCM-X (1.0 g x 100 g-1 x 24 h-1), and (5) SAP + gentamycin (2000 u x 100 g-1 x 24 h-1) + TCM-X (1.0 g x 100 g-1 x 24 h-1) . The medicines were given by way of gastrotube, once every 24 hours, twice in all . Pancreatitis was induced by a single intraperitoneal injection of 500 mg.100 g-1 of L-arginine . Serum endotoxin were observed and the clone forming units from mesenteric lymphnode and pancreas were obtained after 48 hours treatment . 96 hours after the experiment, the bacteria found in the mesenteric lymphnodes and pancreas in groups three, four and five were reduced as compared to that in group two; the levels of serum endotoxin were reduced, too . These data indicate that TCM-X and gentamycin in decontamination by way of gastrotube are effective in preventing bacterial translocation complicated by SAP, and the effect of TCM-X is stronger than that of gentamycin. Oncogene, 2003 Jan 23, 22(3), 370 - 80 Selective ablation of human cancer cells by telomerase-specific adenoviral suicide gene therapy vectors expressing bacterial nitroreductase; Bilsland AE et al.; Reactivation of telomerase maintains telomere function and is considered critical to immortalization in most human cancer cells . Elevation of telomerase expression in cancer cells is highly specific: transcription of both RNA (hTR) and protein (hTERT) components is strongly upregulated in cancer cells relative to normal cells . Therefore, telomerase promoters may be useful in cancer gene therapy by selectively expressing suicide genes in cancer cells and not normal cells . One example of suicide gene therapy is the bacterial nitroreductase (NTR) gene, which bioactivates the prodrug CB1954 into an active cytotoxic alkylating agent . We describe construction of adenovirus vectors harbouring the bacterial NTR gene under control of the hTR or hTERT promoters . Western blot analysis of NTR expression in normal and cancer cells infected with adenoviral vectors showed cancer cell-specific nitroreductase expression . Infection with adenoviral telomerase-NTR constructs in a panel of seven cancer cell lines resulted in up to 18-fold sensitization to the prodrug CB1954, an effect that was retained in two drug-resistant ovarian lines . Importantly, no sensitization was observed with either promoter in any of the four normal cell strains . Finally, an efficacious effect was observed in cervical and ovarian xenograft models following single intratumoural injection with low doses of vector, followed by injection with CB1954. Novartis Found Symp, 2002, 247, 162 - 77; discussion 177-81, 198-206, 244-52 Modelling the bacterial chemotaxis receptor complex; Shimizu TS et al.; The pathway controlling chemotaxis in Escherichia coli is the simplest and most well understood cell signalling system to date . However, quantitative models based on the available data still fail to reproduce important features of the pathway . Most notably, the observed sensitivity of cells to very small changes in stimulus concentrations cannot be reproduced by conventional models based on the measured concentrations, binding affinities and rate constants of the proteins involved . This discrepancy, together with recent experimental findings, drew our attention to the spatial organization of molecules within the cell and in particular to the clusters of receptors localised at the cell poles . A stochastic simulator for chemical reactions, STOCHSIM, was previously developed to model the chemotaxis pathway at the level of individual molecular interactions . This program has now been extended to incorporate a spatial representation that allows the interaction between molecules in a two-dimensional lattice to be simulated . In silico 'experiments' using this new version of STOCHSIM demonstrate that lateral interactions between clustered receptors can significantly enhance the excitation response . The adaptation reactions may also exploit the proximity of receptor molecules, and a hypothetical mechanism by which this may occur is currently being tested. Hua Xi Kou Qiang Yi Xue Za Zhi, 2001 Apr, 19(2), 93 - 4, 98 {A study on bacterial contamination of dental handpieces}; Hu T et al.; OBJECTIVE: The aim of this study is to evaluate the effects of different factors, including concentration of the bacterial suspension, number of stops set on handpieces, and different disinfectants, on bacterial contamination of handpieces and their clinical significance . METHODS: Bacterial contamination of 20 handpieces were analyzed in different concentration of bacterial suspension, number of stops set on handpieces and after using different out surface disinfectants . RESULTS: Statistical increment (P < 0.05) of handpieces inside contamination was observed relevant to increment of the bacterial suspension concentration . No statistical relevance was seen between handpiece inside contamination and the number of stops set on handpieces(P > 0.05) . Disinfection with different disinfectants significantly decreased out surface contamination(P < 0.05), but no statistical change of bacteria (P > 0.05) was observed inside handpieces . CONCLUSION: As the concentration of bacterial suspension is the key factor affecting the inside contamination of handpieces, oral cavity cleanness is essential before dental therapy . The surface disinfection of handpieces is necessary after dental therapy, but cannot prevent cross-contamination between patients. J Am Soc Nephrol, 2003 Feb, 14(2), 317 - 26 Bacterial CpG-DNA aggravates immune complex glomerulonephritis: role of TLR9-mediated expression of chemokines and chemokine receptors; Anders HJ et al.; Immune complex glomerulonephritis (GN) often deteriorates during infection with viruses and bacteria that, in contrast to mammals, have DNA that contains many unmethylated CpG motifs . Balb/c mice with horse apoferritin-induced GN (HAF-GN) were treated with either saline, CpG-oligodeoxynucleotides (ODN), or control GpC-ODN . Only CpG-ODN exacerbated HAF-GN with an increase of glomerular macrophages, which was associated with massive albuminuria and increased renal MCP-1/CCL2, RANTES/CCL5, CCR1, CCR2, and CCR5 mRNA expression . CpG-ODN induced a Th1 response as indicated by serum anti-HAF IgG(2a) titers, mesangial IgG(2a) deposits, and splenocyte IFN-gamma secretion . Messenger RNA for the CpG-DNA receptor Toll-like reeptor 9 (TLR9) was present in kidneys with HAF-GN but not in normal kidneys . The source of TLR9 mRNA in HAF-GN could be infiltrating macrophages or intrinsic renal cells, e.g., mesangial cells; but, in vitro, only murine J774 macrophages expressed TLR9 . In J774 cells, CpG-ODN induced the chemokines MCP-1/CCL2 and RANTES/CCL5 and the chemokine receptors CCR1 and CCR5 . It is concluded that CpG-DNA can aggravate preexisting GN via a shift toward a Th1 response but also by a novel pathway involving TLR9-mediated chemokine and chemokine receptor expression by macrophages, which may contribute to the enhanced glomerular macrophage recruitment and activation . This mechanism may be relevant during infection-triggered exacerbation of human immune-complex GN and other immune-mediated diseases in general. Pediatr Pathol Mol Med, 2002 Nov-Dec, 21(6), 557 - 67 Reduced acute phase response to differentiate between viral and bacterial infections in children; Urbach J et al.; The objective of this study was to document the reduced acute phase response that appears in children with viral as opposed to bacterial infections . The white blood cell count (WBCC), the erythrocyte sedimentation rate (ESR), and leukocyte and erythrocyte adhesiveness/aggregation were determined in 36 children with acute bacterial infection, 29 children with viral infection, and 19 controls . A significant reduced WBCC, ESR, and leukocyte and erythrocyte adhesiveness/aggregation was noted in the children with acute viral infection as opposed to those with bacterial infection: 10,800 +/- 3600 and 20,000 +/- 10,000 cells/cm2, 29 +/- 21 and 53 +/- 35 mm Hg, 23 +/- 9 and 41 +/- 15%, and 3.4 +/- 5.1 and 9.8 +/- 13.6 microns, respectively . The results indicate that a reduced acute phase response can be observed in children with an acute viral infection . This can have diagnostic implications and pathophysiological consequences in terms of less flow impairment in the microcirculation due to less red and white blood cell aggregation. J Am Chem Soc, 2003 Jan 29, 125(4), 1041 - 55 Role of copper ion in bacterial copper amine oxidase: spectroscopic and crystallographic studies of metal-substituted enzymes; Kishishita S et al.; The role of the active site Cu(2+) of phenylethylamine oxidase from Arthrobacter globiformis (AGAO) has been studied by substitution with other divalent cations, where we were able to remove >99.5% of Cu(2+) from the active site . The enzymes reconstituted with Co(2+) and Ni(2+) (Co- and Ni-AGAO) exhibited 2.2 and 0.9% activities, respectively, of the original Cu(2+)-enzyme (Cu-AGAO), but their K(m) values for amine substrate and dioxygen were comparable . X-ray crystal structures of the Co- and Ni-AGAO were solved at 2.0-1.8 A resolution . These structures revealed changes in the metal coordination environment when compared to that of Cu-AGAO . However, the hydrogen-bonding network around the active site involving metal-coordinating and noncoordinating water molecules was preserved . Upon anaerobic mixing of the Cu-, Co-, and Ni-AGAO with amine substrate, the 480 nm absorption band characteristic of the oxidized form of the topaquinone cofactor (TPQ(ox)) disappeared rapidly (< 6 ms), yielding the aminoresorcinol form of the reduced cofactor (TPQ(amr)) . In contrast to the substrate-reduced Cu-AGAO, the semiquinone radical (TPQ(sq)) was not detected in Co- and Ni-AGAO . Further, in the latter, TPQ(amr) reacted reversibly with the product aldehyde to form a species with a lambda(max) at around 350 nm that was assigned as the neutral form of the product Schiff base (TPQ(pim)) . Introduction of dioxygen to the substrate-reduced Co- and Ni-AGAO resulted in the formation of a TPQ-related intermediate absorbing at around 360 nm, which was assigned to the neutral iminoquinone form of the 2e(-)-oxidized cofactor (TPQ(imq)) and which decayed concomitantly with the generation of TPQ(ox) . The rate of TPQ(imq) formation and its subsequent decay in Co- and Ni-AGAO was slow when compared to those of the corresponding reactions in Cu-AGAO . The low catalytic activities of the metal-substituted enzymes are due to the impaired efficiencies of the oxidative half-reaction in the catalytic cycle of amine oxidation . On the basis of these results, we propose that the native Cu(2+) ion has essential roles such as catalyzing the electron transfer between TPQ(amr) and dioxygen, in part by providing a binding site for 1e(-)- and 2e(-)-reduced dioxygen species to be efficiently protonated and released and also preventing the back reaction between the product aldehyde and TPQ(amr). Hua Xi Yi Ke Da Xue Xue Bao, 2001 Sep, 32(3), 452 - 3, 458 {Clindamycin soluble suppository-production and its efficacy in treating bacterial vaginosis}; Huang W et al.; OBJECTIVE: To make the soluble suppository of clindamycin which is an effective drug for treatment of Bacterial vaginosis and to treat patients in clinical trials . METHODS: Outpatients who had vaginal infections from Sept . 1998 to Jan . 1999 were investigated . The diagnoses of bacterial vaginosis were made according to Amsel's criteria and 80 patients were included in this study . They were divided into two groups to be treated respectively by metronidanole suppository and clindamycin suppository which were made by our hospital pharmacy . RESULTS: The cure rate of metronidanole was 87.5% and that of clindamycin 90.3%; No significant difference was noted between the two groups(P > 0.05) . The new clindamycin suppository was not irritant and it was easy to dissolve in vagina . There was no discontinued treatment because of side-effect . CONCLUSION: The clindamycin suppository does not irritate the mucosa of vagina, it is an efficacious remedy for bacterial vaginosis. Appl Microbiol Biotechnol, 2003 Jan, 60(5), 571 - 6 Epub 2002 Oct 24. Expression of bacterial poly(3-hydroxybutyrate) synthesis genes in hairy roots of sugar beet (Beta vulgaris L.); Menzel G et al.; Three genes from Ralstonia eutropha necessary for poly(3-hydroxybutyrate) (PHB) synthesis were introduced into the hairy roots of sugar beet . Transformation of a vector construct harbouring the PHB genes, each fused to the coding region of the pea ribulose-bisphosphate carboxylase plastid targeting sequence, resulted in 20 transgenic hairy-root clones, producing up to 55 mg high molecular PHB/g dry weight, as identified by gas chromatography, gel permeation chromatography and HPLC . Accumulation of PHB polymer in sugar beet root leucoplasts was confirmed by transmission electron microscopy . Thus, for the first time, plastidic PHB production was demonstrated for roots of a carbohydrate-storing crop plant. Mol Microbiol, 2003 Feb, 47(3), 671 - 97 The bacterial linear motor of Spiroplasma melliferum BC3: from single molecules to swimming cells; Trachtenberg S et al.; Spiroplasma melliferum BC3 are wall-less bacteria with internal cytoskeletons . Spiroplasma, Mycoplasma and Acholeplasma belong to the Mollicutes, which represent the smallest, simplest and minimal free-living and self-replicating forms of life . The Mollicutes are motile and chemotactic . Spiroplasma cells are, in addition, helical in shape . Based on data merging, obtained by video dark-field light microscopy of live, swimming helical Spiroplasma cells and by cryoelectron microscopy, unravelling the subcellular structure and molecular organization of the cytoskeleton, we propose a functional model in which the cytoskeleton also acts as a bacterial linear motor enabling and controlling both dynamic helicity and swimming . The cytoskeleton is a flat, monolayered ribbon constructed from seven contractile fibrils (generators) capable of changing their length differentially in a co-ordinated manner . The individual, flat, paired fibrils can be viewed as chains of tetramers approximately 100 A in diameter composed of 59 kDa monomers . The cytoskeletal ribbon is attached to the inner surface of the cell membrane (but is not an integral part of it) and follows the shortest helical line on the coiled cellular tube . We show that Spiroplasma cells can be regarded, at least in some states, as near-perfect dynamic helical tubes . Thus, the analysis of experimental data is reduced to a geometrical problem . The proposed model is based on simple structural elements and functional assumptions: rigid circular rings are threaded on a flexible, helical centreline . The rings maintain their circularity and normality to the centreline at all helical states . An array of peripheral, equidistant axial lines forms a regular cylindrical grid (membrane), by crossing the lines bounding the rings . The axial and peripheral spacing correspond to the tetramer diameter and fibril width (100 A) respectively . Based on electron microscopy data, we assign seven of the axial grid lines in the model to function as contractile generators . The generators are clustered along the shortest helical paths on the cellular coil . In the model, the shortest generator coincides with the shortest helical line . The rest, progressively longer, six generators follow to the right or to the left of the shortest generator in order to generate the maximal range of lengths . A rubbery membrane is stretched over (or represented by) the three-dimensional grid to form a continuous tube . Co-ordinated, differential length changes of the generators induce the membranal cylinder to coil and uncoil reversibly . The switch of helical sense requires equalization of the generators' length, forming a straight cylindrical tube with straight generators . The helical parameters of the cell population, obtained by light microscopy, constitute several subpopulations related, most probably, to cell size and age . The range of molecular dimensions in the active cytoskeleton inferred from light microscopy and modelling agrees with data obtained by direct measurements of subunit images on electron micrographs, scanning transmission electron microscopy (STEM) and diffraction analysis of isolated ribbons . Swimming motility and chemotactic responses are carried out by one or a combination of the following: (i) . reciprocating helical extension and compression ('breathing'); (ii) . propagation of a deformation (kink) along the helical path; (iii) . propagation of a reversal of the helical sense along the cell body; and (iv) . irregular flexing and twitching, which is functionally equivalent to standard bacterial tumbling . Here, we analyse in detail only the first case (from which all the rest are derived), including switching of the helical sense. Bull Exp Biol Med, 2002 Oct, 134(4), 407 - 10 Expression of neuronal and inducible nitric oxide synthases in the thymus under normal conditions and after administration of bacterial endotoxin; Titova IV et al.; The topography of thymocytes expressing neuronal and inducible nitric oxide synthases and changes in the content of luminescent immunoreactive products in these cells after intraperitoneal injection of bacterial lipopolysaccharide were studied by double immunohistochemical labeling . Under normal conditions neuronal nitric oxide synthase-immunopositive cells formed a wide network in thymus medulla (except for perivascular regions) . Inducible nitric oxide synthase was expressed in single cells at the corticomedullary boundary . Lipopolysaccharide markedly increased the intensity of luminescence and number of inducible nitric oxide synthase-immunoreactive cells . However, this agent sharply decreased the intensity of luminescence in neuronal nitric oxide synthase-immunopositive cells of the stroma . Our results indicate that neuronal and inducible nitric oxide synthases are synthesized in various stromal cells of the thymus . Expression of these enzyme isoforms undergoes opposite changes during inflammation. Endocr Res, 2002 Nov, 28(4), 357 - 61 Bacterial lipopolysaccharide directly stimulates cortisol secretion in human adrenal cells; Vakharia K et al.; Adrenomedullin and pro-adrenomedullin N-terminal 20 peptide (PAMP) are expressed in vascular cells and in the adrenal cortex and medulla . Lipopolysaccharide (LPS), a bacterial product that induces septic shock, is a potent stimulant of adrenomedullin secretion in vascular cell types and is also known to stimulate the hypothalamo-pituitary-adrenal axis (HPA) . The present study was designed to investigate the actions of LPS on the human adrenocortical cell line, H295R . Exposure of cells to LPS for 24 hours had no effect on adrenomedullin or PAMP secretion, but was found to significantly and selectively increase cortisol secretion with no effect on aldosterone . Dibutyryl cAMP, however, caused a significant increase in both adrenomedullin and PAMP release over this time period . There are two conclusions which can be drawn from these observations . First that adrenomedullin and PAMP are regulated by different mechanisms in vascular and adrenal cells and second, that LPS is able to directly stimulate cortisol secretion, with implications for the physiological response to septic shock. Cell Mol Life Sci, 2002 Nov, 59(11), 1902 - 13 Bacterial cold-shock proteins; Ermolenko DN et al.; Members of a family of small cold-shock proteins (CSPs) are induced during bacterial cell response to a temperature decrease . Here we review available data about the structure, molecular properties, mechanism of induction and possible functions of CSPs . CSPs preferentially bind single-stranded RNA and DNA and appear to play an important role in cell physiology under both normal and cold-shock conditions . Although the function of CSPs in cold-shock adaptation has not yet been elucidated in detail, a number of experimental evidences suggests that CSPs bind messenger RNA (mRNA) and regulate ribosomal translation, rate of mRNA degradation and termination of transcription. Int J Pharm, 2003 Jan 30, 251(1-2), 155 - 63 Adsorbed pluronics on the skin of human volunteers: effects on bacterial adhesion; Marsh LH et al.; An amphiphilic copolymer, Pluronic F127, has been deposited, by adsorption, to the skin of human volunteers and the ability of the coated skin to resist bacterial colonisation has been evaluated . In parallel, the ability of the same copolymer to act as a bacterial release agent has been evaluated . In both cases, F127 proved to be of little added value in formulations designed to suppress the bacterial colonisation of human skin. Cell, 2003 Jan 10, 112(1), 131 - 40 The bacterial toxin RelE displays codon-specific cleavage of mRNAs in the ribosomal A site; Pedersen K et al.; The Escherichia coli relBE operon encodes a toxin-antitoxin pair, RelE-RelB . RelB can reverse inhibition of protein synthesis by RelE in vivo . We have found that although RelE does not degrade free RNA, it cleaves mRNA in the ribosomal A site with high codon specificity . Among stop codons UAG is cleaved with fast, UAA intermediate and UGA slow rate, while UCG and CAG are cleaved most rapidly among sense codons . We suggest that inhibition of protein synthesis by RelE is reversed with the help of tmRNA, and that RelE plays a regulatory role in bacteria during adaptation to poor growth conditions. Curr Biol, 2003 Jan 8, 13(1), R16 - 8 Bacterial division: the fellowship of the ring; Margolin W; The Z ring, composed of the tubulin homolog FtsZ, is essential for bacterial cell division . Recently a new protein, ZapA, has been discovered that localizes to the Z ring and stabilizes it, probably by promoting the bundling of FtsZ protofilaments. Wei Sheng Yan Jiu, 2000 Jul, 29(4), 207 - 8 {Effects of bacterial protective liquid on radiation biology effect of mouse bone marrow cells}; Xu S et al.; To study the relationship between bacterial protective liquid (BPL) and the biological effect of mouse bone marrow cells by gamma-ray irradiation, apoptosis and the number of CD4+, CD8+ and NK cells were analyzed by flow cytometry(FCM), the results showed that BPL decreased the apoptosis and increased the proliferation of cells(P < 0.01) . The theorem of action was related with raising the percentage vitality of NK cells and the quantity of CD4+ and CD8+ . This study proved that BPL might have protective effect against radiation injury. Toxicol Sci, 2003 Jan, 71(1), 124 - 32 The role of Kupffer cells and TNF-alpha in monocrotaline and bacterial lipopolysaccharide-induced liver injury; Yee SB et al.; Coexposure to small, noninjurious doses of the pyrrolizidine alkaloid phytotoxin monocrotaline (MCT) and bacterial lipopolysaccharide (LPS) results in synergistic hepatotoxicity . Both centrilobular and midzonal liver lesions occur and are similar to those seen from large, toxic doses of MCT and LPS, respectively . The nature of the lesions in vivo and results from studies in vitro suggest that injury is mediated indirectly rather than from a simple interaction of MCT and LPS with hepatic parenchymal cells . Accordingly, the role of inflammatory factors, such as Kupffer cells and TNF-alpha, in the development of MCT/LPS-induced liver injury was investigated . In Sprague-Dawley rats, MCT (100 mg/kg, i.p.) was administered 4 h before LPS (7.4 x 10(6) EU/kg, i.v.) . Pretreatment of these animals with gadolinium chloride, an inhibitor of Kupffer cell function, attenuated liver injury 18 h after MCT administration . An increase in plasma TNF-alpha preceded the onset of hepatic parenchymal cell injury, raising the possibility that this inflammatory cytokine contributes to toxicity . Either pentoxifylline, an inhibitor of cellular TNF-alpha synthesis, or anti-TNF-alpha serum coadministered to MCT/LPS-treated animals significantly attenuated liver injury . These results suggest that Kupffer cells and TNF-alpha are important mediators in the synergistic hepatotoxicity resulting from MCT and LPS coexposure. Nucleic Acids Res, 2003 Jan 1, 31(1), 109 - 13 PACRAT: a database and analysis system for archaeal and bacterial intergenic sequence features; Ray WC et al.; Analysis of intergenic sequences for purposes such as the investigation of transcriptional signals or the identification of small RNA genes is frequently complicated by traditional biological database structures . Genome data is commonly treated as chromosome-length sequence records, detailed by gene calls demarcating subsequences of the chromosomes . Given this model, the determination of non-called subsequences between any gene and its nearest neighbors requires an exhaustive search of all gene calls associated with the chromosome . Further compounding the issue, the location of intergenic regions for many called genes cannot be resolved unambiguously due to uncertainties in gene boundaries, as well as the presence of other conflicting gene calls . To address these difficulties we have constructed the PACRAT database system . PACRAT preprocesses GenBank genome submissions, evaluates for every gene the character of its relationship to those genes nearest to it, and produces a relationally linked model of the gene ordering for the genome . Using this information, the interface allows the researcher to query gene data as well as intergenic sequence data based on a number of criteria . These include the ability to filter searches based on the status of start and stop positions, or upstream/downstream sequences as conflicting with called genes and automated extension of upstream or downstream searches to find probable operon promoters or terminators . The database is also indexed by KEGG classification, allowing, for example, functionally-related groups of high-quality promoter-containing regions to be easily retrieved as a group. J Biol Chem, 2003 Apr 18, 278(16), 13706 - 11 Epub 2003 Jan 07. An iron-dependent bacterial phospholipase D reminiscent of purple acid phosphatases; Zambonelli C et al.; Recombinant phospholipase D (PLD) from Streptomyces chromofuscus (scPLD) has been characterized using colorimetric assays, spectroscopic investigations, and site-directed mutagenesis . scPLD, which shows phosphodiesterase activity toward a wide variety of phospholipids and phosphatase activity toward p-nitrophenyl phosphate, exhibits a visible absorption band with lambda(max) at 570 nm . Metal ion analysis performed by inductively coupled plasma mass spectroscopy shows the presence of approximately 1 equivalent of iron, 0.27 equivalent of manganese, and 0.1 equivalent of zinc per mole of protein as isolated . The metal ion content coupled with the visible absorption feature is compatible with the presence of Fe(3+)-tyrosinate coordination . When scPLD was dialyzed against solutions containing Mn(2+), Zn(2+) or EDTA, the Fe(3+) content was reduced to variable extents, and the residual specific activity correlated well with the residual iron content . Sequence homology with metal ion binding motifs in known alkaline phosphatases and purple acid phosphatase from red kidney bean shows that most of the residues involved in metal ion coordination are conserved among all the sequences considered . Mutation of some of these conserved residues (C123A, D151A, Y154F, and H391A) produced enzymes lacking iron with dramatically reduced PLD activity but little change in secondary structure or ability to bind to small unilamellar vesicles of phosphatidylcholine (with Ba(2+)) or phosphatidic acid . We suggest that scPLD is a member of a family of phosphodiesterase/phosphatases with structural and mechanistic similarity to iron-dependent purple acid phosphatases. Mol Microbiol, 2003 Jan, 47(2), 583 - 93 Thinking and decision making, bacterial style: Bacterial Neural Networks, Obernai, France, 7th-12th June 2002; Armitage JP et al.; Bacteria exhibit a bewildering range of behavioural responses and permutations of metabolic pathways for maximum exploitation of their environment . These are based on sensory perception of external and internal signals through batteries of surface and cytoplasmic receptors, evaluation of complex information flows and rapid decision making . Appreciation of the diversity of bacterial behaviour and adaptation capacities requires the study of a broad range of organisms and at this meeting we sampled more than 30 species with new findings which included the nature of gaseous receptors, advances in chemotaxis, subversion of host defences by pathogens, adaptation to high salt, community life and its obvious benefits, cell to cell communications and even the nature of bacterial circadian rhythms . With around 80 bacterial genomes now completed, and many more almost there, it was appropriate to complete the meeting with an introduction to Systems Biology and prospects for simulating the virtual cell . The versatility and seemingly 'intelligent' behaviour of bacteria will continue to fascinate, and this meeting on Bacterial Neural Networks fully reflected the excitement of this field. SADJ, 2002 Oct, 57(10), 391 - 4 A study of orofacial bacterial infections in elderly Nigerians; Ugboko VI et al.; This retrospective study was designed to determine the pattern of occurrence, and principles of management of orofacial bacterial infections in elderly Nigerians . Ninety-two patients aged 55-90 years (mean +/- SD 62.4 +/- 7.5 years), were admitted into the study (male:female ratio 1.1:1) . The origin of orofacial infection was predominantly odontogenic and the lower face was three times more commonly involved . The percentage of patients with underlying systemic disease was 16.2% . Patients with underlying disease had a 3.5 times greater risk of developing orofacial infections . Similarly, a significant correlation was demonstrated between both sexes of patients with orofacial infections and underlying illness (P < 0.05) . There was a general delay before presenting for treatment (average 19.5 weeks) and the mortality rate was 3.3% . The study findings demonstrate that a significant relationship exists between elderly Nigerians with orofacial infections and systemic disease. J Am Chem Soc, 2003 Jan 15, 125(2), 530 - 5 Direct electrochemistry of a bacterial sulfite dehydrogenase; Aguey-Zinsou KF et al.; Sulfite dehydrogenase from Starkeya novella is an alphabeta heterodimer comprising a 40.6 kDa subunit (containing the Mo cofactor) and a smaller 8.8 kDa heme c subunit . The enzyme catalyses the oxidation of sulfite to sulfate with the natural electron acceptor being cytochrome c550 . Its catalytic mechanism is thought to resemble that found in eukaryotic sulfite oxidases . Using protein film voltammetry and redox potentiometry, we have identified both Mo- and heme-centered redox responses from the enzyme immobilized on a pyrolytic graphite working electrode: E m,8 (Fe III/II) +177 mV; E m,8 (Mo VI/V) +211 mV and E m,8 (Mo V/IV) -118 mV vs NHE; Upon addition of sulfite to the electrochemical cell a steady-state voltammogram is observed and an apparent Michaelis constant (Km) of 26(1) microM was determined for the enzyme immobilized on the working electrode surface, which is comparable with the value obtained from solution assays. J Biol Chem, 2003 Mar 14, 278(11), 9227 - 34 Epub 2003 Jan 03. Identification of the human and bovine ATP:Cob(I)alamin adenosyltransferase cDNAs based on complementation of a bacterial mutant; Leal NA et al.; In humans, deficiencies in coenzyme B12-dependent methylmalonyl-CoA mutase (MCM) lead to methylmalonyl aciduria, a rare disease that is often fatal in newborns . Such deficiencies can result from inborn errors in the MCM structural gene or from mutations that impair the assimilation of dietary cobalamins into coenzyme B12 (Ado-B12), the required cofactor for MCM . ATP:cob(I)alamin adenosyltransferase (ATR) catalyzes the terminal step in the conversion of cobalamins into Ado-B12 . Substantial evidence indicates that inherited defects in this enzyme lead to methylmalonyl aciduria, but the corresponding ATR gene has not been identified . Here we report the identification of the bovine and human ATR cDNAs as well as the corresponding human gene . A bovine liver cDNA expression library was screened for clones that complemented an ATR-deficient bacterial strain for color formation on aldehyde indicator medium, and four positive clones were isolated . The DNA sequences of two clones were determined and found to be identical . Sequence similarity searching was then used to identify a homologous human cDNA (89% identity) and its corresponding gene that is located on chromosome XII . The bovine and human cDNAs were independently cloned and expressed in Escherichia coli . Enzyme assays showed that expression strains produced 87 and 98 nmol/min/mg ATR activity, respectively . These specific activities are in line with values reported previously for bacterial ATR enzymes . Subsequent studies showed that the human cDNA clone complemented an ATR-deficient bacterial mutant for Ado-B12-dependent growth on 1,2-propanediol . This demonstrated that the human ATR is active under physiological conditions albeit in a heterologous host . In addition, Western blots were used to show that ATR expression is altered in cell lines derived from cblB methylmalonyl aciduria patients compared with cell lines from normal individuals . We propose that inborn errors in the human ATR gene identified here result in methylmalonyl aciduria . The identification of genes involved in this disorder will allow improvements in the diagnosis and treatment of this serious disease. J Biol Chem, 2003 Feb 21, 278(8), 5509 - 12 Epub 2003 Jan 04. Host recognition of bacterial muramyl dipeptide mediated through NOD2 . Implications for Crohn's disease; Inohara N et al.; NOD2, a protein associated with susceptibility to Crohn's disease, confers responsiveness to bacterial preparations of lipopolysaccharide and peptidoglycan, but the precise moiety recognized remains elusive . Biochemical and functional analyses identified muramyl dipeptide (MurNAc-L-Ala-D-isoGln) derived from peptidoglycan as the essential structure in bacteria recognized by NOD2 . Replacement of L-Ala for D-Ala or D-isoGln for L-isoGln eliminated the ability of muramyl dipeptide to stimulate NOD2, indicating stereosele