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| Plant J, 1995 Mar, 7(3), 525 - 33 Construction of a rice bacterial artificial chromosome library and identification of clones linked to the Xa-21 disease resistance locus; Wang GL et al.; A bacterial artificial chromosome (BAC) library consisting of 11,000 clones with an average DNA insert size of 125 kb was constructed from rice nuclear DNA . The BAC clones were stable in E . coli after 100 generations of serial growth . Transformation of the BAC clones by electroporation into E . coli was highly efficient and increased with decreasing size of the DNA inserts . The library was evaluated for the presence of organellar, repeated, and telomeric sequences . A very low percentage (< 0.3%) of the library consisted of chloroplast and mitochondrial clones . Eighteen BACs were identified that hybridized with an Arabidopsis telomere repeat . Sixteen BACs hybridized with the AA genome-specific repetitive sequence pOs48 . Twelve clones were isolated that hybridized with three DNA markers linked to the Xa-21 disease resistance locus . The results indicate that the BAC system can be used to clone and manipulate large pieces of plant DNA efficiently. J Clin Microbiol, 1995 Mar, 33(3), 745 - 7 Culture of intestinal biopsy specimens and stool culture for detection of bacterial enteropathogens in patients infected with human immunodeficiency virus . The Berlin Diarrhea/Wasting Syndrome Study Group; Liesenfeld O et al.; The diagnostic yields of stool cultures and biopsy specimens for the detection of enteric bacterial pathogens in 213 human immunodeficiency virus-infected patients were compared . Forty-five percent (19 of 42) of the pathogens were detected exclusively by stool culture, 2% (1 of 42) of the isolates were detected exclusively by culture of biopsy specimens, and 53% (22 of 42) were detected by both methods . Repeated stool cultures remain the most important means of diagnosing enteric bacterial pathogens, which were encountered in 20% (40 of 213) of all patients . The additional culture of biopsy specimens should be reserved for patients with suspected mycobacteriosis. J Endocrinol, 1995 Mar, 144(3), 457 - 62 Differential effect of selective block of alpha 2-adrenoreceptors on plasma levels of tumour necrosis factor-alpha, interleukin-6 and corticosterone induced by bacterial lipopolysaccharide in mice; Hasko G et al.; The effect of selective block of alpha 2-adrenoreceptors on plasma levels of tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and corticosterone induced by bacterial lipopolysaccharide (LPS) was investigated in mice using ELISA and RIA . It was found that the LPS-induced TNF-alpha response was significantly blunted in mice pretreated with CH-38083, a novel and highly selective alpha 2-adrenoreceptor antagonist (the alpha 2/alpha 1 ratio is > 2000) . In contrast, LPS-induced increases in both corticosterone and IL-6 plasma levels were further increased by CH-38083 . Since it has recently been shown that the selective block of alpha 2-adrenoreceptors located on noradrenergic axon terminals resulted in an increase in the release of noradrenaline (NA), both in the central and peripheral nervous systems, and, in our experiments, that propranolol prevented the effect of alpha 2-adrenoreceptor blockade on TNF-alpha plasma levels induced by LPS, it seems likely that the excessive stimulation by NA of beta-adrenoreceptors located on cytokine-secreting immune cells is responsible for this action . Since it is generally accepted that increased production of TNF-alpha is involved in the pathogenesis of inflammation and endotoxin shock on the one hand, and corticosterone and even IL-6 are known to possess anti-inflammatory properties on the other hand, it is suggested that the selective block of alpha 2-adrenoreceptors might be beneficial in the treatment of inflammation and/or endotoxin shock. Infect Agents Dis, 1995 Mar, 4(1), 1 - 12 Processing of bacterial antigens for presentation to class I and II MHC-restricted T lymphocytes; Harding CV et al.; Phagocytosis leads to the destruction of many bacteria and the proteolytic degradation of bacterial antigens within phagolysosomes to produce immunogenic peptides that bind to Class II major histocompatibility (MHC) molecules within vacuolar compartments . On the other hand, Class I MHC molecules bind cytosol-derived peptides, including peptides from bacteria that escape the vacuolar system and penetrate into the cytosol . A recently described pathway may also allow the presentation of peptides from intravacuolar organisms by Class I MHC molecules in some cases . T cell recognition of peptide-MHC complexes then provides the primary basis for specific immunity to protein antigens of bacteria . This article will review the subcellular compartments and mechanisms involved in generating immunogenic peptides, the subcellular localization of MHC molecules that bind these peptides, and bacterial parameters that affect antigen processing. Ann Surg, 1995 Mar, 221(3), 299 - 307 Elemental diet and IV-TPN-induced bacterial translocation is associated with loss of intestinal mucosal barrier function against bacteria; Deitch EA et al.; OBJECTIVE: The goal of the current study was to directly assess the role of loss of mucosal barrier function in nutritionally induced bacterial translocation . BACKGROUND: Parenteral and certain elemental enteral diets have been shown to promote bacterial translocation . The mechanisms underlying this observation, especially the question of whether nutritionally induced bacterial translocation is primarily related to loss of intestinal barrier function, versus an impaired immune system, remain to be fully elucidated . METHODS: Bacterial translocation was measured in vivo, ileal mucosal membranes were harvested, and their electrophysiologic properties and barrier function were measured ex vivo in the Ussing chamber system 7 days after receiving total parenteral nutrition solution parenterally (IV-TPN) or enterally (elemental diet) . Chow-fed rats served as control subjects . RESULTS: The incidence of bacterial translocation was significantly increased both to the mesenteric lymph nodes in vivo and across the in vitro Ussing chamber-mounted ileal mucosal membranes of the elemental diet-fed and IV-TPN-fed rats . The magnitude of Escherichia coli and phenol red transmucosal passage in the Ussing chamber was significantly higher in the IV-TPN-fed rats than in the elemental diet-fed or chow-fed animals . The potential differences across the ileal membrane were similar between the three groups at all time points . However, the specific resistances of the ileal membranes of the IV-TPN and elemental diet groups were significantly less than the chow-fed animals, indicating increased membrane permeability . CONCLUSIONS: Loss of intestinal barrier function plays a major role in nutritionally induced bacterial translocation, and the loss of mucosal barrier function to both E . coli and phenol red appeared greater in the IV-TPN than the elemental diet-fed rats. Arch Dis Child Fetal Neonatal Ed, 1995 Mar, 72(2), F102 - 6 Presence of secretory IgA antibodies to an enteric bacterial pathogen in human milk and saliva; Nathavitharana KA et al.; The concept of a common mucosal immune system in man was tested by examining the concurrent presence of specific-secretory IgA (SIgA) antibodies in human milk and saliva from three groups of subjects: 64 Sri Lankan women living in Sri Lanka; 20 immigrant Asian women living in Birmingham (median duration of residence in the United Kingdom five years); and 75 Caucasian women living in Birmingham (controls) . Enzyme linked immunosorbent assays (ELISA) were developed to detect enterotoxigenic Escherichia coli (ETEC) colonisation factor/1 (CFA/1) specific SIgA antibodies in milk and saliva . ETEC CFA/1 specific SIgA antibody activity was detectable in milk (37.5% and 25%) and saliva (42.1% and 35%) of Sri Lankan and immigrant Asian women, respectively, but not in any of the Caucasian controls . Eighty five point two per cent of subjects who were positive had specific antibodies detectable in both milk and saliva; 5% of all Sri Lankan women and 10% of all immigrant Asian women had detectable antibody only in saliva . These observations lend further strong support to the idea that a common mucosal immune system exists in man . The continuing presence of specific SIgA antibodies in Asian immigrants to previously encountered antigens suggests that there may be an 'immunological memory' in the human secretory immune system. Microbiol Rev, 1995 Mar, 59(1), 48 - 62 Energetics of bacterial growth: balance of anabolic and catabolic reactions; Russell JB et al.; Biomass formation represents one of the most basic aspects of bacterial metabolism . While there is an abundance of information concerning individual reactions that result in cell duplication, there has been surprisingly little information on the bioenergetics of growth . For many years, it was assumed that biomass production (anabolism) was proportional to the amount of ATP which could be derived from energy-yielding pathways (catabolism), but later work showed that the ATP yield (YATP) was not necessarily a constant . Continuous-culture experiments indicated that bacteria utilized ATP for metabolic reactions that were not directly related to growth (maintenance functions) . Mathematical derivations showed that maintenance energy appeared to be a growth rate-independent function of the cell mass and time . Later work, however, showed that maintenance energy alone could not account for all the variations in yield . Because only some of the discrepancy could be explained by the secretion of metabolites (overflow metabolism) or the diversion of catabolism to metabolic pathways which produced less ATP, it appeared that energy-excess cultures had mechanisms of spilling energy . Bacteria have the potential to spill excess ATP in futile enzyme cycles, but there has been little proof that such cycles are significant . Recent work indicated that bacteria can also use futile cycles of potassium, ammonia, and protons through the cell membrane to dissipate ATP either directly or indirectly . The utility of energy spilling in bacteria has been a curiosity . The deprivation of energy from potential competitors is at best a teleological explanation that cannot be easily supported by standard theories of natural selection . The priming of intracellular intermediates for future growth or protection of cells from potentially toxic end products (e.g., methylglyoxal) seems a more plausible explanation. G Ital Cardiol, 1995 Mar, 25(3), 335 - 40 {Reparative surgery of the mitral valve in bacterial endocarditis}; Fucci C et al.; INTRODUCTION . Short and long-term results of valve repair for pure mitral insufficiency resulting from native valve endocarditis are reported in 28 consecutive patients with a mean age of 55 years (range 18-74) . METHODS . Six patients had acute endocarditis, with positive blood cultures in three of them . The mean time between onset of endocarditis symptoms and operation was 23 days in patients with acute endocarditis and 4.6 years in patients with healed endocarditis . Preoperatively, 87% of the patients were in NYHA class III . Indications for operation were heart failure (24 patients) and uncontrolled sepsis (4 patients) . Mitral valvuloplasty was combined with other procedures in 4 patients . There was previous underlying valve pathology in 75% . RESULTS . Mitral repair was performed according to the techniques proposed by Carpentier; in 2 cases we used an original technique consisting of a double-orifice repair . Only one patient died in the hospital (operative mortality: 3.5%) . By actuarial methods 96% of the patients were alive 6 years postoperatively . During the follow-up period there was no recurrence of endocarditis and no reoperation for valvular insufficiency . Ninety-three per cent of the patients were in NYHA class I or II . CONCLUSIONS . We conclude that mitral valve repair for insufficiency resulting from bacterial endocarditis is possible in acute and healed disease, has a low operative mortality and has resulted in patients free of recurrent infection . Mitral valve repair is an attractive alternate to valve replacement in bacterial endocarditis. J Infect, 1995 Mar, 30(2), 89 - 94 Bacterial meningitis: causes for concern . The Research Committee of the BSSI; Bacterial translocation and enteral nutrition in humans: an outsider looks in; GI-Hepatology-Nutrition Section, Department of Veterans Affairs Medical Center, Washington, DC 20422, USAPURPOSE: To assess the literature documenting the existence of bacterial translocation in humans, the effects of enteral nutrition on bacterial translocation in humans, and the hypothesis that enteral nutrition prevents bacterial translocation in humans . DATA IDENTIFICATION: Sources included Medline search, references from review articles, and references from animal and human studies . STUDY SELECTION: The goal was to include all animal and human studies directly addressing questions of bacterial translocation and nutritional status or nutritional support . DATA EXTRACTION: An attempt was made to briefly summarize methodology and findings of relevent studies . No general attempt was made to assess quality of individual studies . RESULTS OF DATA SYNTHESIS: Bacterial translocation is a well documented phenomenon in animal models . Starvation and malnutrition of themselves do not induce bacterial translocation, but may facilitate translocation in the presence of other systemic insults . Parenteral nutrition and many forms of enteral nutrition may induce and/or facilitate bacterial translocation . Chow and certain fiber sources seem protective . Moderate direct and several lines of indirect evidence support the existence of bacterial translocation in humans . There is no direct evidence and questionable indirect evidence suggesting that enteral nutrition prevents or modifies bacterial translocation in humans . CONCLUSIONS: The hypothesis relating enteral nutrition and bacterial translocation in critically ill patients remains attractive, but unproven. Immunol Lett, 1995 Mar, 45(3), 185 - 8 In vitro modulation of preleukemic AKR mouse macrophage function by bacterial immunomodulators; Burek B et al.; Spleen macrophages from 1- and 4-month-old preleukemic AKR mice were stimulated in vitro with the bacterial immunomodulators lipopolysaccharide (LPS), peptidoglycan monomer (PGM) and muramyl dipeptide (MDP), in order to study their migration ability and mitochondrial enzyme activity . Macrophages from 1-month-old AKR mice, characterized by higher functional activity, failed to demonstrate any changes in the parameters studied after in vitro stimulation with the employed compounds . Conversely, the depressed macrophage function, spontaneously developed in 4-month-old AKR mice, most probably related to the preleukemic state, improved significantly and to about the same extent with all three immunomodulators. Int J Urol, 1995 Mar, 2(1), 29 - 32 Transurethral balloon laser hyperthermia for chronic non-bacterial prostatitis: a clinical trial; Suzuki T et al.; Transurethral balloon laser hyperthermia (TUBAL-H) was performed on five patients with chronic non-bacterial prostatitis who failed to respond to conventional treatment administered for more than two years . The prostatic interstitial temperature during treatment was measured and the safety and efficacy of this treatment was assessed . TUBAL-H was performed at a target temperature of 43 degrees C (5 mm depth) and at a laser power of 30 watts with urethral cooling for 20 min in the first three patients and for 30 min in the remaining two patients . The prostatic interstitial temperature at a depth of about 5 mm from the urethral surface ranged from 40.5 to 43.0 degrees C during treatment . The temperature of the urethra ranged from 31.5 to 39 degrees C and that of the rectum remained below 39.5 degrees C . After treatment, no abnormal findings were noted in any of the hematological and biological tests carried out, including prostatic specific antigen . The leukocyte count in expressed prostatic secretions fell to less than five cells per high-power field in four of the five patients after three months . A complete improvement in symptoms was observed in one patient, partial improvement in three, and no improvement in one . Based on these results, TUBAL-H was considered to be safe and a suitable treatment for patients with chronic prostatitis. Br Dent J, 1995 Feb 25, 178(4), 133 - 9 Bacterial proteases in gingival crevicular fluid before and after periodontal treatment; Eley BM et al.; Our recent work has developed specific assays for bacterial dipeptidyl peptidase (DPP) and trypsin-like proteases and we have found them in gingival crevicular fluid (GCF) . The purpose of this study was to determine whether their levels reduce following periodontal treatment of chronic periodontitis patients . The probing depth, probing attachment level, gingival index, gingival bleeding index and plaque index were measured at mesio-buccal sites of molars and premolars in 25 untreated patients . At a second visit GCF was collected on filter paper strips for 30 seconds . GCF volumes were determined with a Periotron and the samples eluted into buffer . The patients than received oral hygiene instruction, supra- and subgingival scaling and other appropriate non-surgical treatment . Four weeks later GCF was collected from the same 16 sites and the clinical parameters were measured again . DPP-like activity was determined fluorometrically with Ala-Pro-AFC at pH 8.0 with and without heating to 60 degrees C for 30 minutes . The heat sensitive portion was taken as bacterial DPP activity . Bacterial trypsin-like protease activity was assayed with Z-Val-Lys-Lys-Arg-AFC at pH 7.0 with 2 mM dithiothreitol and 10 microM Z-Phe-Ala-CH2 . Following treatment there were marked reductions in clinical parameters, enzyme total activities and concentrations . All reductions were statistically significant at patient and site level using either individual patient or pooled patient data . Bacterial proteases appear to reflect the clinical status and may be of value in monitoring chronic inflammatory periodontal disease. Eur J Pharmacol, 1995 Feb 24, 275(1), R1 - 3 Increase in serum NG-hydroxy-L-arginine in rats treated with bacterial lipopolysaccharide; Hecker M et al.; Aortic rings isolated from rats 4 h after an injection i.p . of 30 mg/kg Escherichia coli lipopolysaccharide showed a marked hyporeactivity to noradrenaline . This effect was paralleled by an increase in the level of nitrite/nitrate in the serum of lipopolysaccharide-treated rats, indicative of an enhanced nitric oxide (NO) synthase activity . Most important, however, the serum concentration of the NO synthase intermediate, NG-hydroxy-L-arginine, was also markedly elevated from 3.7 to 15.8 microM . Circulating NG-hydroxy-L-arginine may thus represent a sensitive and specific marker of NO synthase activity in vivo. Biochem Biophys Res Commun, 1995 Feb 15, 207(2), 783 - 9 High-yield bacterial expression, purification, and functional reconstitution of the tricarboxylate transport protein from rat liver mitochondria; Xu Y et al.; The rat liver mitochondrial tricarboxylate transport protein has been overexpressed in E . coli . The expressed transporter, which contains a 21 amino acid N-terminal fusion sequence, accumulates in inclusion bodies . Subsequent extraction of the tricarboxylate transporter from isolated inclusion bodies yields approximately 90 mg of transport protein per liter of E . coli culture at a purity of greater than 90% . Upon incorporation into phospholipid vesicles the purified, overexpressed transporter catalyzes a 1,2,3-benezenetricarboxylate-sensitive citrate/citrate exchange (i.e., the defining reaction of the mitochondrial tricarboxylate transporter) . Kinetic characterization of the reconstituted transporter indicates a Km of 0.37 mM and a Vmax of 101 nmol/min/mg protein . The substrate specificity of the reconstituted, expressed transporter is virtually identical to that of the native transporter . These studies represent the first overexpression of the rat liver mitochondrial tricarboxylate transporter . By providing a large amount of highly-purified, functionally competent transporter this system will now enable a variety of structural studies, including site-directed mutagenesis, which heretofore could not be performed. Biochem J, 1995 Feb 15, 306 ( Pt 1), 225 - 34 Tissue- and cell-specific expression of mouse xanthine oxidoreductase gene in vivo: regulation by bacterial lipopolysaccharide; Kurosaki M et al.; The expression of the xanthine oxidoreductase gene was studied in various mouse organs and tissues, under basal conditions and on treatment with bacterial lipopolysaccharide . Levels of xanthine oxidoreductase protein and mRNA were compared in order to understand the molecular mechanisms regulating the expression of this enzyme system . The highest amounts of xanthine oxidoreductase and the respective mRNA are observed in the duodenum and jejunum, where the protein is present in an unusual form because of a specific proteolytic cleavage of the primary translation product present in all locations . Under basal conditions, multiple tissue-specific mechanisms of xanthine oxidoreductase regulation are evident . Lipopolysaccharide increases enzyme activity in some, but not all tissues, mainly via modulation of the respective transcript, although translational and post-translational mechanisms are also active . In situ hybridization studies on tissue sections obtained from mice under control conditions or with lipopolysaccharide treatment demonstrate that xanthine oxidoreductase is present in hepatocytes, predominantly in the proximal tubules of the kidney, epithelial layer of the gastrointestinal mucosa, the alveolar compartment of the lung, the pulpar region of the spleen and the vascular component of the heart. Proc Natl Acad Sci U S A, 1995 Feb 14, 92(4), 1048 - 52 Purification and characterization of recombinant human p50csk protein-tyrosine kinase from an Escherichia coli expression system overproducing the bacterial chaperones GroES and GroEL; Amrein KE et al.; An Escherichia coli expression system overproducing the bacterial chaperones GroES and GroEL was engineered and has been successfully used to produce large quantities of the recombinant human protein-tyrosine kinase p50csk . The co-overproduction of the two chaperones with p50csk results in increased solubility of the kinase and allows purification of milligram amounts of active enzyme . Analysis of the purified protein by SDS/polyacrylamide gel electrophoresis reveals a single band with an apparent molecular mass of 50 kDa, indicating that recombinant human p50csk has been purified to near homogeneity . The purified enzyme displays tyrosine kinase activity as measured by both autophosphorylation and phosphorylation of exogenous substrates . Biochemical properties, including in vitro substrate specificity and enzymatic characteristics of the enzyme, have been assessed and compared with those of members of the Src family of protein-tyrosine kinases . Results indicate that p50csk and p56lck have different substrate specificities and that p50csk and p60c-src have similar kinetic parameters . The successful production and purification of an enzymatically active form of p50csk will enable further characterization of this important kinase and allow clarification of its physiological role . In addition, the results suggest that the approach described may be generally applicable to improve the solubility of recombinant proteins which otherwise are produced in an insoluble form in E . coli. Adv Ther, 1995 Mar-Apr, 12(2), 139 - 46 Ampicillin/sulbactam versus cefazolin or cefoxitin in the treatment of skin and skin-structure infections of bacterial etiology; Chan JC; This randomized, double-blind study compared 1 g of ampicillin plus 0.5 g of sulbactam with 0.5 g of cefazolin in the treatment of cellulitis and with 1 g of cefoxitin in other skin and skin-structure infections . Study drugs were administered intravenously every 6 hours to 58 hospitalized patients . Each indication was evaluated separately . In cellulitis, ampicillin/sulbactam and cefazolin produced clinical cure or improvement in 100% and 91.7% of patients, respectively; duration of hospitalization was 7.7 and 7.2 days . In other skin and skin-structure infections, results for ampicillin/sulbactam and cefoxitin, respectively, were clinical cure or improvement, 80% and 64.7%; treatment failures, 0% and 11.8%; bacterial eradication, 40% and 53%; and duration of hospitalization, 7.7 and 9.4 days . No unusual or unexpected adverse experiences related to any study drug occurred . One patient treated with ampicillin/sulbactam died of a pulmonary embolism, and 1 patient treated with cefoxitin was discontinued from the study following amputation of an infected foot . These events were not considered drug-related . The treatment groups showed no statistically significant differences in efficacy or safety. J Obstet Gynaecol, 1995 Feb, 21(1), 51 - 5 Detection and identification of amines in bacterial vaginosis; Kubota T et al.; For the purpose of determining an objective indicator of bacterial vaginosis (BV), we tried to detect and to identify amines in the vaginal fluid of patients by using the dansyl method (to detect amine in a solution of water) . Bacteria isolated from these patients were also analyzed . 1) In BV (n: 32) 7 kinds of amines were detected; and the positive rate of detection of the dansyl method was 90.6% . No specific bacteria was found in BV . 2) The positive rate was 80.0% for the patients with trichomonas vaginitis (n: 5) . 3) The positive rate of detection was 20.0% in the candida group (n: 5), and 28.6% in the control group (n: 14) . Those amines detected in other than BV-related vaginitis are presumed not to vaporize . For purposes of diagnosis, it would be better to detect vaporized amines instead of detecting amines dissolved in vaginal fluid. J Trauma, 1995 Feb, 38(2), 223 - 7 Detection of acute bacterial infection within soft tissue injuries using a 99mTc-labeled chemotactic peptide; Fischman AJ et al.; OBJECTIVE: Infection imaging with a 99mTc-labeled chemotactic peptide was evaluated in a rabbit model of Escherichia coli infections in burned tissue . MATERIALS AND METHODS: The peptide was radiolabeled with 99mTc via the hydrazino nicotinamide derivative . Three groups of six animals were studied: (group A) unilateral infected burns; (group B) bilateral burns with unilateral infection; and (group C) uninfected burns . Twenty-four hours after injury, groups A and B were infected, and 8 hours later, all animals were injected with approximately 0.50 mCi of 99mTc-peptide . MEASUREMENTS AND MAIN RESULTS: In groups A and B, excellent images of the infections were obtained at 3 to 4 and 16 to 18 hours after injection of the peptide . At 3 to 4 hours after injection, the target-to-background ratios (T/B) were 3.12 +/- 0.28 for group A and 4.33 +/- 0.61 for group B (p = n.s.) . At 16 to 18 hours, the T/B ratios increased significantly (p < 0.01): group A = 8.10 +/- 1.03 and B = 7.70 +/- 1.25 . The T/B ratio for group C was only slightly greater than unity . CONCLUSIONS: These results indicate that 99mTc-labeled chemotactic peptides are effective radiopharmaceuticals for the rapid detection of focal sites of infection within thermally injured tissues. J Surg Res, 1995 Feb, 58(2), 159 - 64 Glutamine reduces bacterial translocation after small bowel transplantation in cyclosporine-treated rats; Zhang W et al.; Bacterial translocation (BT) of enteric organisms is a major cause of sepsis in patients undergoing small bowel transplantation (SBT) . Cyclosporine (CsA) may be toxic to intestinal epithelium and increase the risk of BT . Glutamine (Gln) is the preferred enterocyte fuel and maintains graft epithelial integrity in experimental SBT . This study determined the effects of CsA on mucosal structure and function of transplanted intestinal isograft and examined whether Gln-enriched diet reversed CsA-induced intestinal toxicity . Thirty-three adult Lewis rats underwent resection of the distal 60% of small bowel and received an orthotopic jejunal isograft . Rats received either elemental diet with 2% Gln or the same diet with balanced nonessential amino acids (non-Gln) by gastrostomy for 10 days . CsA (15 mg/kg, im) or olive oil was injected daily . Rats were assigned to four groups: non-Gln with vehicle, non-Gln with CsA, Gln with vehicle, and Gln with CsA . Mucosal villous height, surface area, crypt depth, 14C glucose absorption, BT to mesenteric lymph nodes (MLN), and body weight change were evaluated . The non-Gln with CsA group had the highest incidence of BT (P < 0.001) . Gln groups had significantly decreased BT (P < 0.01) and increased crypt depth and villous surface area (P < 0.01) when compared to non-Gln groups . Body weight significantly decreased in CsA groups when compared to non-CsA groups (P < 0.01) . These results indicate at CsA significantly decreased body weight and increased BT without decreasing mucosal structure and glucose absorption of intestinal isografts.(ABSTRACT TRUNCATED AT 250 WORDS) J Clin Invest, 1995 Feb, 95(2), 725 - 31 Bacterial lipopolysaccharide-mediated fetal death . Production of a newly recognized form of inducible cyclooxygenase (COX-2) in murine decidua in response to lipopolysaccharide; Silver RM et al.; Maternal infection is a cause of spontaneous abortion and preterm labor in humans, but the pathophysiology is unclear . We hypothesized that eicosanoids play an important role in infection-driven pregnancy loss . To investigate this hypothesis, we administered lipopolysaccharide (LPS) to pregnant C3H/HeN mice and found that LPS administration caused fetal death in a dose-dependent fashion . Pretreatment with indomethacin significantly decreased the proportion of fetal death from 83% to < 25% in mice injected with 10 micrograms of LPS . Also, decidual explants from LPS-treated mice produced significantly more inflammatory eicosanoids, including prostaglandins E2 and F2 alpha and thromboxane B2, than controls . We investigated the regulatory mechanisms responsible for increased decidual prostanoid production in response to LPS . Western and Northern blots demonstrated that decidual protein and mRNA levels of a recently recognized highly inducible form of cyclooxygenase, COX-2, were substantially increased in mice treated with LPS . Induction of COX-2 was rapid: mRNA was detected 30 min after LPS injection . In contrast, another form of cyclooxygenase, COX-1, was only minimally induced in response to LPS . Our data indicate that LPS induces decidual prostanoid production via increased COX-2 expression . Since LPS-mediated fetal death is markedly diminished by pretreatment with indomethacin, COX-2-mediated eicosanoid production is likely a key pathophysiologic event in LPS-mediated fetal death. Hepatology, 1995 Feb, 21(2), 340 - 4 Spontaneous bacterial peritonitis in cirrhotic patients treated using paracentesis or diuretics: results of a randomized study; Sola R et al.; Diuretic treatment in cirrhotic patients with ascites increases ascitic fluid concentration of total protein and complement components, and opsonic activity . These changes are not observed in patients treated with paracentesis . Based on these data it has been suggested that therapeutic paracentesis may be associated with an increased risk of spontaneous bacterial peritonitis (SBP) development . To assess this possibility, 80 cirrhotic patients with tense ascites were randomly allocated in two therapeutic groups: group 1 (40 patients) was treated with total paracentesis associated with plasma volume expansion and group 2 was treated with diuretics . After mobilization of ascites, patients from both groups received diuretics to avoid reaccumulation of ascites; cases that developed tense ascites during follow-up (mean follow-up period, 60 +/- 6 and 55 +/- 4 weeks, respectively) were treated according to initial randomization . Patients from both groups had similar results regarding baseline clinical and standard laboratory data, ascitic fluid concentration of total protein, complement components, and opsonic activity . Sixteen patients (7 from group 1 and 9 from group 2) developed SBP during the study period . The 4-week and 1-year probability of SBP occurrence were 2.5% and 18.6%, respectively, in group 1 patients, and 11.9% and 24%, respectively, in group 2 patients . Therefore, our study indicates that therapeutic paracentesis does not increase the early- and long-term risk of SBP development in cirrhotic patients with tense ascites. Glycoconj J, 1995 Feb, 12(1), 1 - 6 Importance of lectins for the prevention of bacterial infections and cancer metastases; Beuth J et al.; Adhesion of bacteria and of metastasizing tumour cells have much in common, especially the participation of lectins in this process . In the future it might be possible to inhibit the metastatic process and bacterial adhesion by blocking with lectins specific for appropriate (oligo) saccharides or glycoconjugates . Initial clinical trials are very promising. J Hepatol, 1995 Feb, 22(2), 143 - 50 Bacterial expression and purification of hepatitis C virus capsid proteins of different size; Handschuh G et al.; Two capsid sequences of the hepatitis C virus were cloned and expressed in an E . coli system . One sequence (c190) comprised the complete capsid region with 573 nucleotides . The other sequence (c125) spanned 375 5'-nucleotides lacking the hydrophobic 3'-part of the hepatitis C virus capsid gene . A full-length and a truncated construct were chosen, since it is not known whether there is 3'-truncation of the hepatitis C virus capsid during protein maturation similar to the situation in some flaviviridae . The corresponding expression clones 190/4 and 125/4 were constructed by polymerase chain reaction cloning into pQE-vectors . The protein expressed, pc125, which is lacking the hydrophobic carboxyterminus of the full-length capsid protein pc190, showed a stronger signal in western blots using anti-hepatitis C virus/EIAII-positive patient's serum . This could be due to better expression and/or better solubilization of pc125 . The truncated protein pc125 displayed the predicted molecular weight of 19 kD, whereas the full-length protein pc190 migrated faster than expected . This could be due to intracellular proteolytic processing, giving rise to a truncated protein or to an atypical mobility in SDS-PAGE gels caused by the hydrophobic nature of the full-length protein . Both proteins were synthesized with an aminoterminal tag of six histidines that could be used for purification by Nickel chelate affinity chromatography . The elution fractions of the two proteins showed additional bands in western blots . Most of these proteins had a mass between 2 and 16 kD and are likely to be degradation products . Protein pc125 could be purified in larger quantities than pc190.(ABSTRACT TRUNCATED AT 250 WORDS) Mol Microbiol, 1995 Feb, 15(4), 601 - 6 The twisted 'life' of DNA in the cell: bacterial topoisomerases; Luttinger A; DNA topoisomerases are essential to the cell for the regulation of DNA supercoiling levels and for chromosome decatenation . The proposed mechanisms for these reactions are essentially the same, except that a change in supercoiling is due to an intramolecular event, while decatenation requires an intermolecular event . The characterized bacterial topoisomerases appear capable of both types of reaction in vitro . Four DNA topoisomerases have been identified in Escherichia coli . Topoisomerase I, gyrase, and topoisomerase IV normally appear to have distinct essential functions within the cell . Gyrase and topoisomerase I are responsible for the regulation of DNA supercoiling . Both gyrase and topoisomerase IV are necessary for chromosomal decatenation . Multiple topoisomerases with distinct functions may give the cell more precise control over DNA topology by allowing tighter regulation of the principal enzymatic activities of these different proteins. Eur J Surg, 1995 Feb, 161(2), 97 - 101 Tumour necrosis factor-alpha and interleukin-6: early indicators of bacterial infection after human orthotopic liver transplantation; Sautner T et al.; OBJECTIVE: To see if it was possible to predict the development of infection after liver transplantation from concentrations of endotoxin, tumour necrosis factor-alpha (TNF-alpha), or interleukin-6 (IL-6) in plasma . DESIGN: Prospective open study . SETTING: University hospital, Austria . SUBJECTS: 46 Consecutive patients who underwent liver transplantation for end stage liver disease, 1989-90 . INTERVENTIONS: Samples of 4 ml blood were taken in endotoxin free tubes, and of 10 ml into heparinised tubes at the beginning of the operation, during hepatectomy, at the beginning and end of the anhepatic phase, 10 minutes after reperfusion, and at the end of the operation . MAIN OUTCOME MEASURES: Correlation between development of infections postoperatively and operative release of endotoxin, TNF-alpha, and IL-6 . RESULTS: There was no correlation between development of postoperative infections and operative concentrations of endotoxin, and of TNF-alpha and IL-6 up to the end of the anhepatic phase . There was, however, a sixfold increase in TNF-alpha and IL-6 concentrations between the end of the anhepatic phase and the end of the operation in patients who subsequently developed infections (p = 0.01) . CONCLUSION: The increase in the concentrations of these two cytokines in the blood after reperfusion of the transplanted liver seems to predict the development of subsequent bacterial infection. J Ind Microbiol, 1995 Feb, 14(2), 61 - 75 Bacterial resistance mechanisms for heavy metals of environmental concern; Ji G et al.; Bacterial species have genetically-determined systems for resistances to toxic heavy metals . Those for metals of environmental concern including mercury cadmium, arsenic and others are briefly summarized, considering the genes of the systems and the biochemical mechanisms by which the resistance proteins function. J Ind Microbiol, 1995 Feb, 14(2), 159 - 63 Bacterial reduction of hexavalent chromium; Wang YT et al.; Cr(VI)-reducing bacteria are widespread and Cr(VI) reduction occurs under both aerobic and anaerobic conditions . Under aerobic conditions, both NADH and endogenous cell reserves may serve as the electron donor for Cr(VI) reduction . Under anaerobic conditions, electron transport systems containing cytochromes appear to be involved in Cr(VI) reduction . High cell densities are necessary to obtain a significant rate of Cr(VI) reduction . Cr(VI) reduction by bacteria may be inhibited by Cr(VI), oxygen, heavy metals, and phenolic compounds . The optimum pH and temperature observed for Cr(VI) reduction generally coincide with the optimal growth conditions of cells . The optimum redox potential for Cr(VI) reduction has not yet been established. Shock, 1995 Feb, 3(2), 116 - 24 Alterations in mucosal morphology and permeability, but no bacterial or endotoxin translocation takes place after intestinal ischemia and early reperfusion in pigs; Schlichting E et al.; Ischemia and reperfusion of the gut may be an important etiological factor in the development of multiple organ failure . We have used a hemorrhagic and a superior mesenteric artery (SMA) occlusion shock model in pigs to estimate the effect of ischemia and reperfusion on intestinal morphology, mucosal permeability, and the occurrence of bacterial or endotoxin translocation . Mucosal ulceration and necrosis were found in the SMA shock model, while the morphological changes were less pronounced in the hemorrhagic shock model . Scanning electron microscopy showed shrinkage of the villi and plugging of the colonic crypts in both shock models . Enterocyte cell kinetics was investigated using 5-bromo-2'-deoksyuridine (BrdU) incorporation and immunovisualization by anti-BrdU antibodies . Cell renewal was almost completely lost from the jejunum to the rectum in both shock models . Intramucosal pH was measured using a tonometer placed in the terminal ileum . Segments of intestinal mucosa were mounted in Ussing chambers, and permeability was measured using radiolabeled probe molecules of differing molecular weights . Augmented molecular flux of inulin (M(r) 5.000) and mannitol (M(r) 182) and loss of short circuit current (Isc) and transepithelial potential difference (PD) were found in mucosae from both shock models . Endotoxin was demonstrated in the ascitic fluid in both shock models; 9.5 (2.7-14.3) (median and 95% confidence interval) EU/mL in the SMA occlusion model and 16.0 (4.9-29.4) EU/mL in the hemorrhagic shock model), but the levels were not significantly higher than in the control model 6.5 (4.3-34.0) EU/mL.(ABSTRACT TRUNCATED AT 250 WORDS) Pediatr Nephrol, 1995 Feb, 9(1), 33 - 5 Secondary amyloidosis from long-standing bacterial endocarditis; Herbert MA et al.; Survival of patients with increasingly complex congenital heart disease has produced a population of children and adolescents who are susceptible to subacute bacterial endocarditis (SBE) . We report a child whose endocarditis went unrecognised, and who developed amyloidosis . Asymptomatic proteinuria, haematuria and renal impairment are occasionally seen in SBE and usually indicate glomerulonephritis . Amyloidosis should also be suspected in children with long-standing bacterial endocarditis with proteinuria or other evidence of renal impairment, especially if associated with organomegaly . The diagnosis is made by renal biopsy. Hematol Oncol Clin North Am, 1995 Feb, 9(1), 167 - 85 Transfusion-transmitted bacterial infection; Krishnan LA et al.; Transfusion-transmitted bacterial infection is a persistent but often underemphasized problem facing transfusion medicine today . The present status of bacterial contamination of red cells, platelet products and plasma, frequency of contamination, and types of organisms implicated is reviewed . Current methods of prevention and detection are discussed, as well as typical clinical presentations and therapy. Int J Biomed Comput, 1995 Feb, 38(2), 131 - 40 Bacterial colony counting using distance transform; Mukherjee DP et al.; A distance-transform based technique is presented for the segmentation of monochrome images of colonies grown on membrane filters . This is used to count the number of Escherichia coli in a given water sample, which is used as a parameter for determining water quality . The result is compared with fuzzy c-means clustering approach. Baillieres Clin Rheumatol, 1995 Feb, 9(1), 179 - 91 Bacterial infections: the arthritis of leprosy; Gibson T; Arthritis is a common feature of leprosy and contributes to disability . Direct invasion of joints and bones by mycobacteria may lead to a destructive arthritis in lepromatous disease . The infective process may involve few or many joints . Reactional states may occur spontaneously but usually after the initiation of anti-mycobacterial treatment . In both the type 1 reaction of borderline case and the type 2 reaction of the lepromatous disease, intense inflammation may occur at sites of infection . The immunology of the reactions is different but they share clinical features including a polyarthritis which may resemble rheumatoid disease . The joint disease may be chronic or relapsing, affecting the wrists and small joints of the hands in particular . Radiological erosions may occur . Mycobacterium leprae is not found in the synovium in this pattern of arthritis . Further study of this phenomenon might yield useful information above the mechanism of joint inflammation in other rheumatic diseases. Baillieres Clin Rheumatol, 1995 Feb, 9(1), 151 - 9 Bacterial infections: bone and joint tuberculosis; Jellis JE; In many countries of the world, there is now a dual epidemic of tuberculosis and HIV disease . HIV specifically eliminates the tissue macrophages and CD4 lymphocytes, the very cells that provide immunity against tuberculosis . Tuberculosis is one of the more virulent opportunistic infections and it therefore appears fairly early in HIV disease . For the same reasons, bone and joint tuberculosis is becoming much more common . The disturbances of the lymphocyte count, ESR and antigen skin tests associated with HIV, now often make tuberculosis difficult to distinguish from other inflammatory lesions . The only change in the pattern of disease that we have so far registered is an increased incidence of disease affecting the lumbar spine . HIV-positive patients respond poorly to chemotherapy and are subject to drug sensitivity reactions . Major surgery is fraught with infectious complications and should be avoided . Once Pott's paraplegia has developed, the demise of the HIV-positive patient is rapid. J Endod, 1995 Feb, 21(2), 70 - 3 Bacterial invasion into dentinal tubules of human vital and nonvital teeth; Nagaoka S et al.; The difference in resistance to bacterial invasion into the dentinal tubules between vital and nonvital teeth has not been determined . This study was conducted to clarify the effect of vital pulp on bacterial invasion into the dentinal tubules . The specimens were 19 intact pairs of bilateral upper third molars of 19 healthy, young adult male volunteers . In each case, 30 or 150 days before extraction, pulpectomies and root canal fillings were carried out unilaterally and a class V cavity involving the dentin was made on the palatal surface of both the pulpectomized tooth and the nonpulpectomized opposite tooth . The cavities were left unprotected to expose them to oral flora until the extractions were done, and the extracted teeth were examined histologically . When extraction followed 150-day exposure to the oral flora, there was a statistically significant difference in the bacterial invasion rate between the vital and nonvital teeth . It was postulated that vital teeth were much more resistant to bacterial invasion into the dentinal tubules than were nonvital teeth, thereby suggesting that the vital pulp plays some important role in this process. J Endod, 1995 Feb, 21(2), 62 - 4 Ability of bacterial endotoxin to diffuse through human dentin; Nissan R et al.; An in vitro system was developed to determine whether bacterial endotoxin is capable of diffusing through dentin without the use of filtration pressure . Cavities were prepared in five third molar teeth in order to produce a split chamber device consisting of occlusal and pulpal chambers with 0.5 mm of intervening dentin . Endotoxin was introduced into the occlusal chamber and the effluent in the pulpal chamber was sampled every 30 min for 5 h and at 24 h using the limulus lysate assay . In four specimens the initial appearance of endotoxin in the effluent ranged from 15 min to 4 1/2 h . In two specimens the concentration of endotoxin in the effluent leveled off in 4 1/2 and 5 h, respectively, whereas in another two the concentration continued to increase throughout the experiment . In one specimen no endotoxin was detected . The results indicate that endotoxin is capable of passing through 0.5 mm of dentin. Br J Rheumatol, 1995 Feb, 34(2), 107 - 12 Defective spontaneous and bacterial lipopolysaccharide-stimulated production of interleukin-1 receptor antagonist by polymorphonuclear neutrophils of patients with active systemic lupus erythematosus; Hsieh SC et al.; Interleukin-1 receptor antagonist (IL-1ra) binds competitively to IL-1 receptors but does not transduce the signal which blocks the biological activities induced by IL-1 . In this study, polymorphonuclear neutrophils (PMN) and mononuclear cells (MNC) from the patients with active systemic lupus erythematosus (SLE) (n = 11), inactive SLE (n = 13) and normal individuals (n = 13) were compared for the IL-1ra producing capacity of these cells . PMN and MNC at a concentration of 1 x 10(6) cells/ml were incubated with medium alone (spontaneous) or stimulated with lipopolysaccharide (LPS, 100 ng/ml) for 24 h . The IL-1ra concentration in the supernatants was quantified by ELISA method . Both spontaneous and LPS-stimulated production of IL-1ra by PMN, but not by MNC, of active SLE were significantly lower than that of inactive SLE or normal groups . Prednisolone (1 and 5 micrograms/ml) did not change the production of IL-1ra by normal PMN either spontaneously or LPS-stimulation in in vitro study . Moreover, the IL-1ra producing capacity of PMN in seven active SLE on admission and after intensive immunosuppressive treatment was measured . These results suggest that the defective IL-1ra production by SLE-PMN is relevant to disease activity and may be regarded as a new indicator of disease activity in patients with active SLE. J Mol Evol, 1995 Feb, 40(2), 136 - 54 Response regulators of bacterial signal transduction systems: selective domain shuffling during evolution; Pao GM et al.; Response regulators of bacterial sensory transduction systems generally consist of receiver module domains covalently linked to effector domains . The effector domains include DNA binding and/or catalytic units that are regulated by sensor kinase-catalyzed aspartyl phosphorylation within their receiver modules . Most receiver modules are associated with three distinct families of DNA binding domains, but some are associated with other types of DNA binding domains, with methylated chemotaxis protein (MCP) demethylases, or with sensor kinases . A few exist as independent entities which regulate their target systems by noncovalent interactions . In this study the molecular phylogenies of the receiver modules and effector domains of 49 fully sequenced response regulators and their homologues were determined . The three major, evolutionarily distinct, DNA binding domains found in response regulators were evaluated for their phylogenetic relatedness, and the phylogenetic trees obtained for these domains were compared with those for the receiver modules . Members of one family (family 1) of DNA binding domains are linked to large ATPase domains which usually function cooperatively in the activation of E . coli sigma 54-dependent promoters or their equivalents in other bacteria . Members of a second family (family 2) always function in conjunction with the E . coli sigma 70 or its equivalent in other bacteria . A third family of DNA binding domains (family 3) functions by an uncharacterized mechanism involving more than one sigma factor . These three domain families utilize distinct helix-turn-helix motifs for DNA binding . The phylogenetic tree of the receiver modules revealed three major and several minor clusters of these domains . The three major receiver module clusters (clusters 1, 2, and 3) generally function with the three major families of DNA binding domains (families 1, 2, and 3, respectively) to comprise three classes of response regulators (classes 1, 2, and 3), although several exceptions exist . The minor clusters of receiver modules were usually, but not always, associated with other types of effector domains . Finally, several receiver modules did not fit into a cluster . It was concluded that receiver modules usually diverged from common ancestral protein domains together with the corresponding effector domains, although domain shuffling, due to intragenic splicing and fusion, must have occurred during the evolution of some of these proteins . Multiple sequence alignments of the 49 receiver modules and their various types of effector domains, together with other homologous domains, allowed definition of regions of striking sequence similarity and degrees of conservation of specific residues . Sequence data were correlated with structure/function when such information was available.(ABSTRACT TRUNCATED AT 250 WORDS) Biophys J, 1995 Feb, 68(2), 708 - 22 A model of excitation and adaptation in bacterial chemotaxis; Hauri DC et al.; We present a model of the chemotactic mechanism of Escherichia coli that exhibits both initial excitation and eventual complete adaptation to any and all levels of stimulus ("exact" adaptation) . In setting up the reaction network, we use only known interactions and experimentally determined cytosolic concentrations . Whenever possible, rate coefficients are first assigned experimentally measured values; second, we permit some variation in these rate coefficients by using a multiple-well optimization technique and incremental adjustment to obtain values that are sufficient to engender initial response to stimuli (excitation) and an eventual return of behavior to baseline (adaptation) . The predictions of the model are similar to the observed behavior of wild-type bacteria in regard to the time scale of excitation in the presence of both attractant and repellent . The model predicts a weaker response to attractant than that observed experimentally, and the time scale of adaptation does not depend as strongly upon stimulant concentration as does that for wild-type bacteria . The mechanism responsible for long-term adaptation is local rather than global: on addition of a repellent or attractant, the receptor types not sensitive to that attractant or repellent do not change their average methylation level in the long term, although transient changes do occur . By carrying out a phenomenological simulation of bacterial chemotaxis, we find that the model is insufficiently sensitive to effect taxis in a gradient of attractant . However, by arbitrarily increasing the sensitivity of the motor to the tumble effector (phosphorylated CheY), we can obtain chemotactic behavior. Biochem Mol Med, 1995 Feb, 54(1), 26 - 32 The human and mammalian N-acetylmuramyl-L-alanine amidase: distribution, action on different bacterial peptidoglycans, and comparison with the human lysozyme activities; Vanderwinkel E et al.; N-acetylmuramyl-L-alanine amidase (EC 3.5.1.28) specifically hydrolyzes the bacterial cell wall peptidoglycans (or mureins) and the muropeptides . The enzyme splits these molecules into two parts: the peptide subunits and the glycan strands or moieties . The bacterial peptidoglycans and their derived muropeptides display a number of biological properties . Removal of the glycosidic part of these molecules abolishes their beneficial as well as their detrimental properties . We report the high level of enzymatic activity found in all mammalian (including human) sera tested . The enzyme also occurred in human saliva, milk, cerebrospinal fluid, and synovial liquid . Mucosal tissue from different parts of the mammalian digestive tract exhibited enzymatic activity, but the enzyme was not detectable in the lumen content . The range of substrate specificity of the human enzyme was evaluated by measuring its action on the peptidoglycans extracted from several bacterial strains and representing different chemotypes and structures . Time course of the muramylalanine amidase and of the lysozyme (both of human origin) activities on some of these peptidoglycans are also reported, with the enzymes acting separately or together . From these data, we would speculate that a probable physiological role of the muramylalanine amidase is the maintenance of adequate ratios between the biologically active muropeptides and their inactive derivatives in the organism, the amidase activity antagonizing the production of biologically active molecules by lysozyme. Philos Trans R Soc Lond B Biol Sci, 1995 Jan 30, 347(1319), 97 - 103 Editing DNA replication and recombination by mismatch repair: from bacterial genetics to mechanisms of predisposition to cancer in humans; Radman M et al.; A hereditary form of colon cancer, hereditary non-polyposis colon cancer (HNPCC), is characterized by high instability of short repeated sequences known as microsatellites . Because the genes controlling microsatellite stability were known in bacteria and yeast, as was their evolutionary conservation, the search for human genes responsible for HNPCC became a 'targeted' search for known sequences . Mismatch-repair deficiency in bacteria and yeast produces multiple phenotypes as a result of its dual involvement in the editing of both replication errors and recombination intermediates . In addition, mismatch-repair functions are specialized in eukaryotes, characterized by specific mitotic (versus meiotic) functions, and nuclear (versus mitochondrial) localization . Given the number of phenotypes observed so far, we predict other links between mismatch-repair deficiency and human genetic disorders . For example, a similar type of sequence instability has been found in HNPCC tumours and in a number of neuro-muscular genetic disorders . Several human mitochondrial disorders display genomic instabilities reminiscent of yeast mitochondrial mismatch-repair mutants . In general, the process of mismatch repair is responsible for the constant maintenance of genome stability and its faithful transmission from one generation to the next . However, without genetic alteration, species would not be able to adapt to changing environments . It appears that nature has developed both negative and positive controls for genetic diversity . In bacteria, for example, an inducible system (sos) exists which generates genetic alterations in response to environmental stress (e.g . radiation, chemicals, starvation) . Hence, the cost of generating diversity to adapt to changing conditions might be paid as sporadic gene alterations associated with disease. Nucleic Acids Res, 1995 Jan 25, 23(2), 269 - 76 Identification of a cDNA for SSRP1, an HMG-box protein, by interaction with the c-Myc oncoprotein in a novel bacterial expression screen; Bunker CA et al.; We describe a system for screening cDNA expression libraries in Escherichia coli based on protein-protein interactions . The system utilizes fusion proteins containing the DNA binding domain of the lambda phage cl repressor and a heterologous dimerization domain, which is the target of the screen . Such chimeric proteins were functional as transcriptional repressors in E.coli; function was dependent on the presence of the heterologous dimerization domain, and function of the chimeras was disrupted by expression of excess dimerization domain . A screen was designed to identify factors that could interact with the heterologous dimerization domain and thereby inactivate the chimeric repressor . We used this screen to identify factors that could interact with the basic helix-loop-helix/leucine zipper domains of c-Myc, and isolated the cDNA for a previously characterized HMG domain protein that interacts specifically with c-Myc in this system . This screening method could be used with proteins that have the ability to homo- or heterodimerize. Biochemistry, 1995 Jan 24, 34(3), 1076 - 83 New inhibitors of the quinol oxidation sites of bacterial cytochromes bo and bd; Meunier B et al.; A screen has been performed of possible inhibitors of the quinol oxidation sites of the two terminal oxidases of Escherichia coli, cytochromes bo and bd . Aurachin C and its analogues were found to be particularly effective inhibitors of both enzymes, whereas aurachin D and its analogues displayed a selectivity for inhibition of cytochrome bd . In addition, a tridecyl derivative of stigmatellin was found to inhibit cytochrome bo at concentrations which were without significant effect on cytochrome bd . Titration of membrane-bound cytochromes bo and bd with aurachin C gave an observed dissociation constant in the range of 10(-8) M . A similar observed dissociation constant was determined for aurachin D inhibition of cytochrome bd . For both enzymes, their kinetic behavior during a series of substrate pulses indicates that it is reduction of the enzyme by quinol, and not reaction with oxygen, which is inhibited . It is concluded that the aurachins are powerful inhibitors of the quinol oxidation sites of bacterial cytochromes bo and bd . The effects of aurachin C on cytochrome bo were investigated in more detail . The number of inhibitor binding sites on the purified enzyme was determined by titration to be 0.6 per enzyme . At an inhibitor/oxidase ratio of 1.0, electron donation into the enzyme from added quinol is extremely slow, making it very unlikely that there is more than one quinone-reactive site . Aurachin C caused a potent inhibition of electron donation from a pulse of quinol.(ABSTRACT TRUNCATED AT 250 WORDS) Anal Biochem, 1995 Jan 20, 224(2), 564 - 71 Assembly of high-resolution bacterial artificial chromosome, P1-derived artificial chromosome, and cosmid contigs; Ashworth LK et al.; The generation of contiguous physical maps is often complicated by a variety of factors including the type of cloning system used . Here we describe procedures for the isolation, rapid characterization, and physical mapping of large-insert recombinant bacterial clones from total human genomic BAC (bacterial artificial chromosome) and PAC (P1-derived artificial chromosome) libraries containing clones with an average insert size of 150 kbp . After initial isolation, the clones were subjected to a variety of fingerprinting procedures including inter-Alu PCR, semiautomated fluorescent finger-printing, and EcoRI restriction fragment mapping . Individual BAC and PAC clones were also used as probes to interrogate arrayed chromosome 19-specific cosmid libraries . The combination of analyses facilitated the identification of chromosome-specific large-insert clones as well as the construction of a large (1.2 Mb) high-resolution BAC, PAC, and cosmid contig in 19q13.2, spanning the region from the carcinoembryonic antigen gene family to the X-ray repair cross complementing 1 DNA repair gene . This type of approach directly demonstrates the utility of large-insert recombinant bacterial clones for the construction of contiguous physical maps of entire chromosomes. Anal Chem, 1995 Jan 15, 67(2), 466 - 71 Improvement in the long-term stability of an amperometric glucose sensor system by introducing a cellulose membrane of bacterial origin; Ammon HP et al.; Classical amperometric glucose sensors that use cellulose membranes of wood origin (Cuprophan) suffer from the fact that their long-term stability in blood is short; therefore, their clinical use is limited . In the present study, a classical amperometric glucose sensor was covered with a bacterial cellulose (BC) membrane . Its surface in comparison to that of the classical glucose sensor (Cuprophan) and its long-term stability were tested in vitro and in vivo . The surface element composition was approximately 44% oxygen and approximately 56% carbon in both membranes and thus typical for cellulose . BC membranes exhibited fiber structure, whereas cup membranes did not . There was also a qualitative difference in protein adsorption between both membranes on exposure to bovine serum albumin . Treatment with Trogamid of one site of the BC membranes allowed linear glucose detection between 0 and 40 mM . Hemocompatibility of BC membranes was improved in comparison to cup membranes on the basis of complement activation (C3a and C5a) . In diluted blood (1:10), the BC-covered sensor exhibited a long-term stability of more than 200 h; in undiluted blood it was stable for about 24 h, which is about 6-7 times longer than the stability of the classical Cup membrane-covered sensor . In in vivo studies, where the BC membrane-covered sensors were connected to the jugular vein of rats, blood glucose levels could be monitored for at least 24 h . In summary, the use of a modified bacterial cellulose membrane to cover the classical amperometric glucose sensor significantly improves the sensor's long-term stability both in vitro and in vivo. J Biol Chem, 1995 Jan 13, 270(2), 581 - 8 The formation of the 2',5'-phosphodiester linkage in the cDNA priming reaction by bacterial reverse transcriptase in a cell-free system; Shimamoto T et al.; Bacterial reverse transcriptase (RT) is responsible for synthesis of multicopy single-stranded DNA (msDNA) consisting of single-stranded DNA linked to an internal guanosine residue of RNA by an unusual 2',5'-phosphodiester linkage . Here we purified a bacterial RT to homogeneity from Escherichia coli harboring the RT gene from retron-Ec73 . The purified RT-Ec73 was able to synthesize msDNA in a cell-free system using an RNA template produced in vitro by T7 RNA polymerase . The in vitro synthesized msDNA was released from the template RNA only when treated with yeast debranching enzyme DBR1, a specific nuclease for a 2',5'-phosphodiester linkage . The position of the branching G residue in the template RNA and the DNA sequence of the cell-free product were identical to those of msDNA-Ec73 synthesized in vivo . These results clearly demonstrate that the formation of the 2',5'-phosphodiester linkage in msDNA synthesis is carried out by RT itself. Nucleic Acids Res, 1995 Jan 11, 23(1), 103 - 8 A bacterial methyltransferase M.EcoHK311 requires two proteins for in vitro methylation; Lee KF et al.; The genes encoding EcoHK311 restriction-modification (R-M) system were isolated from a clinically-isolated Escherichia coli strain HK31 . The entire R-M system of EcoHK311 is located in a 2.1 kb fragment . R.EcoHK311 is an isoschizomer of Eael which recognizes and cleaves Y decreases GGCCR . M.EcoHK31l consists of two polypeptides alpha and beta with sizes 309 and 176 aa, respectively . Polypeptide beta is encoded within aa, alternative reading frame of polypeptide alpha . All the conserved motifs in mC5-MTases can be found in polypeptide alpha except motif IX which is present in polypeptide beta . Polypeptides alpha and beta were separately synthesized in a T7 promoter controlled over-expression system and in vitro methylation occurred only when the two extracts were mixed and thus confirms that two polypeptides are required for methylation. Biochim Biophys Acta, 1995 Jan 2, 1260(1), 14 - 20 Baculovirus expression of human basic fibroblast growth factor from a synthetic gene: role of the Kozak consensus and comparison with bacterial expression; Hills D et al.; Synthetic genes encoding the 146 and 155 amino acid forms of human basic fibroblast growth factor (bFGF) were constructed with codon usage biased towards the polyhedrin-encoding gene of Autographa californica nuclear polyhedrosis virus (AcNPV) . Expression of both bFGF genes in Spodoptera frugiperda (SF-21) suspension cell culture using a recombinant baculovirus yielded approximately 2.5 mg of mitogenically fully active protein per 10(9) cells following heparin-affinity chromatography . To improve translational efficiency, the Kozak consensus sequence was introduced and it was found that neither the replacement of a pyrimidine by a purine at position -3, nor the nature of the base at position +4 had any noticeable effect on the final levels of bFGF expression in SF-21 cells . The bases at these critical points in the consensus do not therefore play a major role in expression levels of the bFGF synthetic genes . The two synthetic genes were also expressed in Escherichia coli as native proteins using the T7 expression system . 5 mg of mitogenically fully active bFGF were obtained from 1 l of bacterial culture . Both insect cell- and E . coli-derived bFGF were equally mitogenic for Swiss 3T3 fibroblasts. J Inflamm, 1995-96, 47(4), 173 - 9 Contribution of TNF/TNF receptor and of Fas ligand to toxicity in murine models of endotoxemia and bacterial peritonitis; Heumann D et al.; Fas/Fas ligand and TNF/TNF receptors are involved in apoptosis . Whether both systems are involved in septic shock has not been determined so far . We investigated the role of TNF/TNFR and Fas/Fas ligand in models of endotoxemia and of speticemia in mice . Upon LPS challenge, TNF and TNFR p55 were involved in the process inducing lethality . FasL did not contribute to enhance lethality, as evidenced in gld mice, lacing FasL . Following an intraperitoneal injection of live E . coli, TNF and TNFR p55 were necessary to combat infection . Disruption of either gene was associated with enhanced lethality and failure to clear the bacteria . No effect observed in gld mice in this peritonitis model . Thus, these observations confirmed the pathogenic role of TNF/TNFR in endotoxemia and its beneficial role in local bacterial infections . In addition the data ruled out a major role for Fas/FasL in septic shock in mice. Mol Biol Rep, 1995-96, 22(2-3), 115 - 23 Analysis of the tertiary structure of bacterial RNase P RNA; Harris ME et al.; The ubiquitous occurrence of ribonuclease P (RNase P) as a ribonucleoprotein and the catalytic properties of bacterial RNase P RNAs indicate that RNA fulfills an ancient and important role in the function of this enzyme . This review focuses on efforts to determine the structure of the bacterial RNase P RNA ribozyme . Phylogenetic comparative analysis of a library of bacterial RNase P RNA sequences has resulted in a well-developed secondary structure model and allowed identification of some elements of tertiary structure . The native structure has been redesigned by circular permutation to facilitate intra- and inter-molecular crosslinking experiments in order to gain further structural information . The crosslinking constraints, together with the constraints provided by comparative analyses, have been incorporated into a first-order model of the structure of of the ribozyme-substrate complex . The developing structural perspective allows the design of self-cleaving pre-tRNA-RNase P RNA conjugates which are useful tools for additional structure-probing experiments. Przegl Lek, 1995, 52(10), 509 - 12 {Acid-base balance and electrolytes in cerebrospinal fluid of patients with bacterial and lymphocytic meningitis}; Wiczkowski A et al.; Acid-base balance and electrolytes concentration in cerebrospinal fluid (CSF) of patients with bacterial and lymphocytic meningitis were assessed . Inflammatory process causing the damage of blood-brain barrier and brains hypoxia leads to statistically significant changes of pH, pO2, bicarbonates and K+ concentrations in CSF of patients with bacterial meningitis, which in lymphocytic meningitis were not observed . Patients with fatal; outcome of bacterial meningitis showed higher CSF's acidosis and lover bicarbonates with higher K+ concentrations, which suggest deeper damage of brain hemostasis regulating mechanisms in those patients. Crit Rev Immunol, 1995, 15(3-4), 317 - 48 The mucosal adjuvant activities of ADP-ribosylating bacterial enterotoxins; Snider DP; The bacterial enterotoxins, cholera toxin and the heat labile toxin of E . coli, are well known adjuvants for mucosal immune response . Their common A chain mediates the toxigenic mechanism by causing ADP ribosylation of G proteins and subsequent elevation of cAMP in target cells . A large IgA and IgG antibody response to admixed protein antigen (Ag) is the hallmark of these adjuvants and is clearly associated with the A chain activity . Expansion of Ag-specific B and T cells, alteration of T cell cytokine production, and changes in regulatory T cells have been reported as adjuvant mechanisms . The B chain derivatives of these toxins can also weakly enhance immune response, especially if covalently associated with Ag and used for nasophyrangeal immunization . Importantly, these toxins or their B chain derivatives can alter the normal immune regulation that produces oral tolerance . This indicates that they modulate mechanisms operative between the mucosal and systemic immune systems . There are some discrepancies between in vitro models of CT or LT activity and in vivo manifestations of their adjuvant activities . Interpretation of current data regarding in vivo mechanism is hampered by an incomplete understanding of how mucosal B and T cells can interact with systemic lymphoid tissue and vice versa . More important, there is no clear understanding of the early effects of the toxins on the local (and draining) mucosal lymphoid tissues . This is especially true in the critical areas of antigen presentation, T and B cell activation, and cytokine production. Virus Genes, 1995, 11(2-3), 95 - 104 Bacterial reverse transcriptase and msDNA; Rice SA et al.; Retrons are a new class of genetic elements found in the chromosome of a large number of different bacteria . These elements code for a reverse transcriptase (RT) that is structurally similar to the polymerases of retroviruses . The retron associated RT is responsible for the production of an unusual extrachromosomal satellite DNA, known as multicopy, single-stranded DNA (msDNA) . Synthesis of msDNA is dependent on a novel self-priming mechanism, resulting in the formation of a 2',5'-phosphodiester bond . A comparison of bacterial RTs is presented, noting conserved and unique features of these polymerases . In addition, the origin, means of dissemination, and possible activities of these functionally obscure retroelements are discussed. Biochimie, 1995, 77(11), 854 - 60 The mouse Kin-17 gene codes for a new protein involved in DNA transactions and is akin to the bacterial RecA protein; Tissier A et al.; We have sought to characterize the molecular basis of the sensitivity to ionising radiation and to identify the genes involved in the cellular response of mammalian cells to such radiation . Using the Escherichia coli model, we tested the hypothesis that functional domains of RecA protein are represented in proteins of mammalian cells . We review here the results obtained in the detection of nuclear proteins of mammalian cells that are recognized by anti-RecA antibodies . We have called them kin proteins . Kin proteins likely play a role in DNA metabolism . We summarize the cloning of the mouse Kin-17 cDNA and our work on the identification and preliminary characterisation of the biochemical properties of mouse kin17 protein, a new nuclear protein able to recognize bent DNA and suspected to be involved in illegitimate recombination . We briefly describe our latest experiments on the molecular characterisation of the mouse Kin-17 gene . Finally, we discuss the properties of kin17 protein and the possible participation of kin17 protein in DNA transactions like transcription or recombination. J Clin Gastroenterol, 1995, 21 Suppl 1, S146 - 50 Tissue IgA antibody against Helicobacter pylori in patients with gastroduodenal diseases: comparison with bacterial culture, serum IgG antibody, and {13C}Urea breath test; Matsukura N et al.; Helicobacter pylori infection is often associated with gastrointestinal diseases, such as chronic gastritis, peptic ulcer, and gastric cancer . After total gastrectomy, positive to negative seroconversion of the H . pylori IgG antibody assayed by enzyme-linked immunosorbent assay (ELISA) was found in 10/15 patients (67%) an average of 8.5 months after surgery . Therefore, the IgG antibody persists for a long time after total removal of the stomach in about 30% of patients . Immunoglobulin A (IgA) is a major component of the local immunity of the stomach mucosa and has a short half-life . Therefore, tissue H . pylori IgA antibodies in biopsy specimens from patients with various gastric diseases were assayed by ELISA and compared with the bacterial culture, serum IgG antibody (ELISA), and {13C}urea breath test results from 144, 170, and 123 endoscopic examinations, respectively . Positivity and negativity of tissue H . pylori IgA coincided with the culture results in 67% of the examinations, and positive IgA antibody but negative culture results were found in 23% . The coincidence of tissue IgA and serum IgG antibodies against H . pylori was 64% and that of negative tissue IgA but positive serum IgG antibody results was 36% . Positivity and negativity of tissue H . pylori IgA antibody coincided with the {13C}urea breath test results in 72% . One month after completion of treatment of peptic ulcer patients for H . pylori infection with lansoprazole and benexate HCl betadex plus amoxicillin, 6/9 (67%) patients showed positive to negative conversion of the tissue IgA antibody, in contrast to no IgG antibody seroconversion . In conclusion, the tissue H . pylori IgA antibody assay is useful for detection of local immunity against H . pylori in the stomach and during follow-up after treatment. Folia Microbiol (Praha), 1995, 40(5), 462 - 66 Mutagenicity of cytostatic drugs in a bacterial system . II . DNA-repair test; Marhan J; A liquid micromethod modification of the DNA-repair test using an automatic growth analyzer was developed . The wild strain Escherichia coli WP2 and six repair-deficient isogenic test strains were used . To compare this repair test with the conventional plate Ames test, a set of nine cytostatics were tested . Cloturin, adriamycin, mitoxantron, oracin, lomustine and tris(2-chloroethyl)amine were found to be positive, and 6-mercaptopurine, butocin and cyclophosphamide negative . The experimental micromethod appears to be useful for assessing the differential lethality in bacteria and can be combined in any short-term test system to predict genotoxicity. Klin Med (Mosk), 1995, 73(6), 30 - 1 {Pathogenesis of cerebral edema-swelling in bacterial meningoencephalitis}; Iarosh OA; Nuclear-absorption spectrometry was used to measure concentration of some metals in cerebrospinal liquid (CSL) of 70 patients with bacterial meningoencephalitis and in different brain compartments of 10 dead patients . Estimation of the difference between dry and wet tissue mass provided the picture of watering for different brain parts . A correlation analysis demonstrated a quantitative relationship between metals and water in different brain compartments . Quantitative determination of CSL metals (potassium, sodium, magnesium, manganese, copper, zinc) enables diagnosis of brain swelling and prognosis of the inflammation outcome. Ann Biol Clin (Paris), 1995, 53(7-8), 429 - 34 {Helpful to bacterial identification: an alternative to probability calculation}; Van Oystaeyen B; In the field of bacterial identification, the usual probabilistic approach may be criticized to some extent for the way it deals with weak tests . To improve this, we propose a modification which consists to compare the probability of the observed profile to the average of all the possible answers of a given bacteria . The result is discussed and showed to be understood in terms of plausibility rather than probability. Biochimie, 1995, 77(7-8), 677 - 94 Electron and proton transfer to the quinones in bacterial photosynthetic reaction centers: insight from combined approaches of molecular genetics and biophysics; Sebban P et al.; We present here new results together with an overview of the current knowledge on the coupled processes of electron and proton transfer in bacterial reaction centers . The importance of a multidisciplinary approach associating molecular genetics, structural biology, biochemistry and spectroscopy is underlined . We emphasize the electrostatic role of the protein to maintain a negative electrostatic potential near the second quinone electron acceptor in order to: i) accelerate the overall rate of proton transfer from the cytoplasm to this acceptor by increasing the pKs of some groups involved in this process; ii) increase the local proton concentration near this acceptor . We also point out the possibility of long distance propagation of the electrostatic effects through the protein associated with relaxation processes triggered by the formation of the semiquinone anions on the first flash. Scand J Infect Dis, 1995, 27(5), 431 - 4 Adjunctive corticosteroid therapy in bacterial meningitis; Lauritsen A et al.; Experimental studies of bacterial meningitis have shown that components of bacterial cell walls stimulate the local production of inflammatory cytokines in the cerebrospinal fluid, leading to inflammation and alterations in the cerebral microvasculature . Animal studies and clinical trials have demonstrated that adjunctive corticosteroid therapy reduces the production of cytokines in the CSF . This results in decreased severity of the inflammatory process and fewer neurologic sequelae . These data support the use of adjunctive dexamethasone in infants and children with S . pneumoniae and H . influenzae type B (HiB) meningitis . There is not sufficient evidence supporting the use of adjunctive corticosteroid therapy in children with meningitis caused by N . meningitidis . Also, the routine use of dexamethasone in adult meningitis cannot presently be recommended . When using dexamethasone timing is crucial . Administration before or with antibiotics is optimal for attenuating the subarachnoid space inflammatory response . Patients receiving the therapy need careful monitoring for the possibility of gastrointestinal bleeding . Future studies of the pathogenesis and pathophysiology of bacterial meningitis may lead to the development of other adjunctive treatment strategies, improving the outcome of this serious disease. Microbiol Immunol, 1995, 39(9), 715 - 23 Inhibition by bacterial lipopolysaccharide of spontaneous and TNF-alpha-induced human neutrophil apoptosis in vitro; Hachiya O et al.; In the previous paper (Takeda et al, Int . Immunol., 5, 691-694, 1993), we demonstrated that tumor necrosis factor-alpha (TNF-alpha) promptly accelerates apoptosis of human neutrophils in vitro . In order to determine the role of neutrophil apoptosis in defending against bacterial infection, we studied the effect of bacterial lipopolysaccharide (LPS) on this process . LPS inhibited spontaneous and TNF-alpha-induced human neutrophil apoptosis in vitro, as determined by 1) light and electron microscopy, 2) flow cytometry, and 3) agarose gel electrophoresis of DNA . Low concentrations of cycloheximide, a protein synthesis inhibitor, which alone did not affect neutrophil apoptosis, were able to reduce spontaneous apoptosis inhibition by LPS, suggesting the involvement of newly synthesized protein in this phenomenon. Bol Chil Parasitol, 1995 Jan-Jun, 50(1-2), 37 - 41 {Hydatidosis of the humerus complicated with fracture, bacterial infection, fistula, and extraosseous localization}; Sapunar J et al.; The clinical case of an apparently healthy 63-year-old man from a rural area, with previous contact with dogs, who had a pathological fracture of the right humerus is presented . Initially he presented slight local pain, and functional discapacity . Eight months later, after a radiological study and surgery (curettage), diagnosis of hydatid disease was made . Later on, after receiving two courses with albendazole, the parent continued in similar conditions for seven years, when his situation became complicated with bacterial, fistula and extraoseous hydatidosis . The humerus was resected and a segmentary prothesis was successfully set. Acta Microbiol Immunol Hung, 1995, 42(3), 261 - 9 Interaction of immunomodulant substances in mouse experiments with special regard to drug sensitivity and bacterial translocation; Anderlik P et al.; In acute toxicity experiments changes in drug sensitivity and in the rate of bacterial translocation (BT) were investigated in mice treated with immunomodulatory drugs: dianhydrogalactitol (DAG) in doses 20 and 30 mg/kg, chlorpromazine (CPZ) in doses 60 and 75 mg/kg and Mannozym (M) in dose equivalent to 40 mg per kg zymosan . The drugs were used separately or in combination . The sensitivity of mice to immunosuppressive DAG or CPZ was higher in the case of combined treatment than that of separately treated ones . The rate of BT was also higher in mice receiving combined treatment . Pretreatment with M exerting an immunostimulatory effect, influenced neither the sensitivity of mice to DAG or CPZ nor the very low normal rate of BT . The present results reinforced the authors' earlier observations that the effects of immunosuppressive drugs cumulated in and caused more serious damage of the organism . The increase in drug sensitivity to immunosuppressive agents may be connected with an increased rate of BT and effect of endotoxin. Hum Mutat, 1995, 6(3), 226 - 31 Comparison between medium-chain acyl-CoA dehydrogenase mutant proteins overexpressed in bacterial and mammalian cells; Jensen TG et al.; Medium-chain acyl-CoA dehydrogenase (MCAD) deficiency is a potentially lethal inherited defect in the beta-oxidation of fatty acids . By comparing the behaviour of five missense MCAD mutant proteins expressed in COS cells and in Escherichia coli, we can define some of these as "pure folding mutants." Upon expression in E . coli, these mutant proteins produce activity levels in the range of the wild-type enzyme only if the chaperonins GroESL are co-overproduced . When overexpressed in COS cells, the pure folding mutants display enzyme activities comparable to the wild-type enzyme . The results suggest that the MCAD mutations can be modulated by chaperones, a phenomenon that may influence the manifestation of the MCAD disease. Prog Clin Biol Res, 1995, 392, 209 - 18 The role of bacterial translocation on Kupffer cell immune function following hemorrhage; Chaudry IH et al.; Studies have shown that Kupffer cell and splenic macrophage, as well as peritoneal macrophage antigen presentation function, was significantly depressed following hemorrhage and remained so for at least 96 hours after resuscitation . Although macrophage antigen presentation was depressed, in all the cell populations studied, it was only the Kupffer cells which were upregulated to produce increased inflammatory cytokines . Furthermore, Kupffer cells from hemorrhaged animals exhibited enhanced, as opposed to reduced toxicity by peritoneal and splenic macrophages . This correlated well with increased cell-associated TNF on Kupffer cells . as well as increased capacity of Kupffer cells to release inflammatory cytokines after hemorrhage . It, therefore, could be postulated that while the enhanced Kupffer cell cytotoxicity may be beneficial in the destruction of pathogens seen in the liver due to bacterial translocation, this same activity may also contribute directly or indirectly to hepatocellular dysfunction and injury which is seen following hemorrhagic shock . Nonetheless, the depression in various immune functions after hemorrhage and resuscitation was comparable in both endotoxin-tolerant and -intolerant mice . Thus, it is debatable whether the alterations in immune function seen after hemorrhage are primarily due to the release of endotoxin into the blood stream during and/or following the hemorrhagic insult . Although translocation and/or endotoxemia occurs following severe hemorrhage, endotoxin may not be the sole or primary agent responsible for the induction of immunodepression after hemorrhage . The depressed Kupffer cell functions and increased inflammatory cytokine release by these cells can be significantly improved by post-treatment of animals with chloroquine, ibuprofen, diltiazem or ATP-MgCl2 . Thus, these agents offer new therapeutic modalities in restoring the depressed Kupffer cell immune functions and in the treatment of generalized immunosuppression, as well as for decreasing the susceptibility to sepsis which is observed following severe blood loss. Crit Care Med, 1995 Jan, 23(1), 125 - 31 Calcium and phospholipase A2 appear to be involved in the pathogenesis of hemorrhagic shock-induced mucosal injury and bacterial translocation; Xu D et al.; OBJECTIVE: The mechanism by which hemorrhagic shock injures the gut and leads to the translocation of bacteria remains incompletely determined . Since increased free cellular calcium levels and phospholipase A2 activity can lead to cellular injury and both have been documented in certain shock states, the hypothesis that calcium or phospholipase A2 may play a role in hemorrhagic shock-induced gut mucosal injury and bacterial translocation was tested . DESIGN: Prospective animal study with concurrent controls . SETTING: Small animal laboratory . SUBJECTS: Fifty-seven male Sprague-Dawley rats weighing 250 to 350 g . INTERVENTIONS: Five groups of rats were tested utilizing a nonlethal hemorrhagic shock model (mean arterial pressure of 30 mm Hg for 30 mins) . These groups included: a) sham-shock, b) shock, c) shock plus quinacrine (inhibitor of phospholipase A2), d) shock plus diltiazem (calcium-channel blocker) administered 5 mins before hemorrhage, and e) shock plus diltiazem administered at the end of shock period and before resuscitation . At 24 hrs postshock or sham-shock, the animals were killed, the mesenteric lymph node and cecum were cultured and the gut was examined histologically . MEASUREMENTS AND MAIN RESULTS: The occurrence rate of shock-induced bacterial translocation (90%) was significantly reduced in rats receiving quinacrine (27%) or preshock diltiazem (21%) (p < .05), but not postshock diltiazem (63%) . Bacterial translocation did not occur in sham-shocked rats . The same amount of blood withdrawal was needed between all groups of rats to induce and maintain shock . Quinacrine and diltiazem administration largely prevented shock-induced ileal and cecal mucosal injury . CONCLUSIONS: The observation that quinacrine and preshock diltiazem limited the extent of shock-induced mucosal injury and bacterial translocation indicate that calcium and phospholipase A2 are involved in the pathogenesis of shock-induced mucosal injury and bacterial translocation . The fact that preshock but not postshock diltiazem was protective indicates that the process leading to shock-induced calcium-mediated tissue injury and bacterial translocation was initiated during the ischemic rather than the reperfusion period . However, since neither quinacrine nor diltiazem was fully protective, other factors, such as oxidants, are also likely to be involved in the pathogenesis of shock-induced mucosal injury and bacterial translocation. Klin Padiatr, 1995 Jan-Feb, 207(1), 12 - 6 {Hearing disorders in children less than 16 months of age after bacterial meningitis with reference to cerebrospinal fluid elastase}; Wilken B et al.; Hearing impairment as a sequela of acute bacterial meningitis is a well known complication . Dexamethasone therapy in addition to antibiotics is beneficial in the reduction of deafness, implicating that inflammation may be one reason for hearing impairment . The risk of hearing impairment in different types of bacterial meningitis is well studied . In very young children < 1.5 years of life the incidence of hearing loss and the possible correlation of laboratory data with the development of deafness is yet unknown . We therefore examined the brainstem auditory evoked potentials in 25 children between the first month and the 16th month of life who we treated for meningitis during 3 years in our hospital . 11 children were treated with dexamethasone . In 9 children we found abnormal brainstem auditory evoked potentials, which we controlled every 3 months . 7 children had transient conductive hearing impairment with good recovery during the first year after the disease . In 2 cases we found permanent bilateral sensorineural hearing loss . There was a significant relationship between hearing loss and elastase in cerebrospinal fluid . Dexamethasone reduced this relationship . A screening of hearing should be performed as routine control in all patients with acute meningitis . The association of high elastase in cerebrospinal fluid and later hearing impairment indicates a pathophysiological relation between activation of granulocytes and hearing loss. Acta Obstet Gynecol Scand, 1995 Jan, 74(1), 67 - 70 Detection of bacterial vaginosis in wet mount, Papanicolaou stained vaginal smears and in gram stained smears; Platz-Christensen JJ et al.; In a prospective study of 107 women, bacterial vaginosis was clinically diagnosed in 34 women . Compared with clinical diagnosis of bacterial vaginosis, detection of clue cells in Papanicolaou stained vaginal smears showed a sensitivity of 88.2%, a specificity of 98.6%, a positive predictive value of 96.8% and a negative predictive value of 94.7% . The corresponding values for detection of bacterial vaginosis in Gram stained smears compared with the clinical diagnosis were 100%, 97.3%, 94.4% and 100%, respectively . Compared with clue cells in wet smears, identification of clue cells in Papanicolaou stained vaginal smears showed a Kappa index of 0.87 and compared with Gram stain criteria a Kappa index of 0.94 . The correlation between Gram stain and Papanicolaou stained vaginal smears showed a Kappa index of 0.89 . In contrast to the results of earlier investigators our studies indicate that the demonstration of clue cells in Papanicolaou stained vaginal smears correlate reasonably well with the conventional clinical criteria . However, the Gram stain method may be more reliable than the Papanicolaou method. Am J Physiol, 1995 Jan, 268(1 Pt 1), L144 - 51 Interaction of surfactant protein A with bacterial lipopolysaccharide may affect some biological functions; Kalina M et al.; Cultured alveolar type II cells and alveolar macrophages were found to secrete colony-stimulating factors (CSF) into the medium . Surfactant protein A (SP-A; 0.1-5 micrograms/ml) and bacterial lipopolysaccharide (LPS; 10-20 micrograms/ml) were found to upregulate the secretion of CSF (seven-fold) from these cells . However, a reversal of the stimulatory effect was observed when the two agents were added simultaneously to the cells . SP-A-enhanced phagocytosis of bacteria by alveolar macrophages was also inhibited by simultaneous addition of SP-A and LPS . Thus some biological activities attributed to either SP-A or LPS are inhibited in the simultaneous presence of the two agents . We therefore investigated the possibility of interaction and binding between SP-A and LPS molecules . Our biochemical data that include immunoblots and enzyme-linked immunosorbent assay support the notion that SP-A is capable of binding LPS, and this interaction is time and concentration dependent . The binding was partially inhibited (60%) by antibody to SP-A . The binding was calcium independent and was not affected by excess carbohydrates such as methyl alpha-D-mannopyranoside or heparin . Lipid A, the hydrophobic component of LPS, however, inhibited the SP-A-LPS interaction and also caused a partial reversal of the binding . Thus these results indicate that lipid A is associated with this binding . The biological implication of SP-A-LPS interaction, especially during inflammatory responses, is discussed. J Leukoc Biol, 1995 Jan, 57(1), 174 - 9 Macrophages derived from C3H/HeJ (Lpsd) mice respond to bacterial lipopolysaccharide by activating NF-kappa B; Ding A et al.; The effects of bacterial lipopolysaccharide (LPS) on macrophage gene expression are mediated in part by its ability to induce activation of transcription factor NF-kappa B . We compared the ability of LPS-treated macrophages from Lpsn (LPS-responsive) C3H/HeN and Lpsd (LPS-hyporesponsive) C3H/HeJ mice to mobilize NF-kappa B by electrophoretic mobility shift assays with oligonucleotide probes containing a unique NF-kappa B sequence from the promoter of inducible nitric oxide synthase (iNOS) . In response to ng/ml concentrations of LPS, this probe bound proteins that appeared rapidly in the nuclei of thioglycollate-elicited macrophages and bone marrow-derived macrophage cell lines from both Lpsn and Lpsd mice . Only in macrophages from Lpsn mice, however, was LPS able to induce iNOS or tumor necrosis factor alpha . NF-kappa B-containing DNA-protein complexes from Lpsd macrophages were formed in lesser amounts than from Lpsn macrophages but shared the same composition, insofar as they displayed the same electrophoretic mobilities and content of heterodimers of p50/RelA (p65) and p50/c-rel . Two conclusions emerge from these findings: (1) NF-kappa B activity alone is not sufficient for induction of certain LPS-responsive genes and (2) An LPS-response pathway involving activation of NF-kappa B is preserved in Lpsd mice . The inability of cells from Lpsd mice to induce gene expression in response to LPS thus cannot be attributed to inability to activate NF-kappa B. Dig Dis Sci, 1995 Jan, 40(1), 183 - 5 Bacterial esophagitis associated with CD4+ T-lymphocytopenia without HIV infection . Possible role of corticosteroid treatment; Richert SM et al.; Although infectious esophagitis is usually due to infection with Candida, herpes virus, or cytomegalovirus, bacterial esophagitis is occasionally observed . Recently, patients have been reported with CD4+ T-lymphocytopenia without HIV infection . Bacterial esophagitis per se has not been reported in these patients . We report the case of an 80-year-old patient admitted with a COPD exacerbation after being on chronic steroids . The patient developed esophageal symptoms and was found to have bacterial esophagitis by biopsy . Her CD4+ counts were found to be low, but she denied HIV risk factors and HIV testing was negative . Her CD4+ counts rose into the normal range as her steroids were tapered, and her esophagitis improved on antibiotics . This case is reported to alert physicians to the possible association of bacterial esophagitis with CD4+ T-lymphocytopenia without HIV infection and to discuss the possible etiological role of corticosteroid treatment. Nephrol Dial Transplant, 1995, 10(3), 377 - 81 Risk factors for bacterial infections in chronic haemodialysis adult patients: a multicentre prospective survey; Hoen B et al.; All adult patients from 13 dialysis centres were prospectively followed up for 6 months in an attempt to appraise the current risk factors for bacterial infections in stable chronically haemodialysed patients . Parameters recorded as potential risk factors for BI were age, gender, cause of renal failure, time elapsed since the start of dialysis, history of transplantation, recent surgical procedure, previous bacterial infection, current immunosuppressive or erythropoietin therapy, type of angioaccess device, and serum ferritin level . Six hundred and seven patients (mean age 56.5 years, range 18-85) were enrolled in the study . Mean time elapsed since the start of dialysis was 4.7 years . One hundred and eighteen patients had developed at least one bacterial infection during the study period whereas 489 had remained free of bacterial infection at the end of the follow-up . In multivariate analysis three parameters were found to be significant and independent risk factors for bacterial infection: previous history of bacterial infection (at least one versus no previous episode), type of angioaccess device (catheter versus native fistula), and elevated serum ferritin level (greater versus lower than 500 micrograms/l) . These results support the evidence that impaired host defences in chronic haemodialysis patients may be secondary to the dialysis procedure and suggest that the incidence of bacterial infection in these patients may be further reduced by appropriate supportive therapy. Life Sci, 1995, 56(26), 2319 - 30 Agmatine, the bacterial amine, is widely distributed in mammalian tissues; Raasch W et al.; We sought to determine whether agmatine (decarboxylated arginine), a bacterial product recently discovered for the first time in mammalian brain, was contained in other organs . A method was developed for isolation of agmatine from tissue and detection by RP-HPLC following solid-liquid extraction and derivatization with o-phthalaldehyde and mercaptoethanol . Recovery was about 80% and the limit of fluorometric detection was about 10 pg on column . In male Sprague-Dawley rats agmatine was unevenly and widely distributed in many tissues confirming its presence in mammals . The highest concentration (approximately 71 ng/mg net weight) was found in stomach, with aorta and small intestine next, followed by smaller levels in spleen, adrenal, aorta, and skeletal muscle and brain . Serum concentrations were high . Agmatine in male Long Evans rats of 3, 12, and 24 months of age demonstrated similar but not identical tissue distribution without any effect of aging . Since agmatine binds to alpha 2-adrenergic and imidazoline receptors, is bioactive in a number of tissues, is contained in neurons and is found in serum and tissues, the findings are consistent with a potential role for agmatine as a neurotransmitter and/or hormone . It also raises the possibility that agmatine may, as in bacteria, serve as a polyamine precursor along metabolic pathways previously not detected in mammals. Scand J Gastroenterol Suppl, 1995, 208, 53 - 7 Malnutrition in the developing and developed world: is small intestinal bacterial colonization important? Riordan SM, Bolin TD. Malnutrition is a major problem on a global basis and will continue if current world population trends persist . Small intestinal bacterial colonization may contribute to malnutrition in populations whose dietary intake is marginal, such as those in developing countries and many elderly in the developed world . This review focuses on the evidence for small intestinal bacterial colonization in these subjects and briefly revises current thinking on the pathogenesis and diagnosis of this condition. Minerva Ginecol, 1995 Jan-Feb, 47(1-2), 5 - 8 {Bacterial vaginosis during chemotherapy for gynecological neoplasms . A therapeutic proposal}; Carta G et al.; Bacterial vaginosis (BV) is a well characterized disease, defined Amsel's criteria, whose incidence is significantly increased during chemotherapy for gynecologic malignancies, as previously reported by the authors . Nine patients out of 14 (64%) who were receiving adjuvant chemotherapy for gynecologic tumors showed a clinical and laboratory evidence of bacterial vaginosis and were treated with cetyltrimethilammonium naproxenate vaginal douches . Results were analyzed up to the completion of oncologic chemotherapy: a high efficacy and tolerability for the topical treatment was demonstrated in all cases. Optom Clin, 1995, 4(3), 53 - 64 Diagnosis, laboratory analysis, and treatment of bacterial corneal ulcers; Stonecipher KG et al.; This paper describes the diagnosis, analysis, and management of ophthalmic bacterial corneal infections, with emphasis on the various risk factors, culture techniques, relations to other pathologic states, prophylaxis, sensitivity trends, and treatment protocols associated with bacterial corneal ulcers . New antiinfective agents and their role in the management of current bacterial ophthalmic infections are also discussed. Mol Microbiol, 1995 Jan, 15(1), 13 - 23 Bacterial transposases and retroviral integrases; Polard P et al.; Transposable genetic elements have adopted two major strategies for their displacement from one site to another within and between genomes . One involves passage through an RNA intermediate prior to synthesis of a DNA copy while the other is limited uniquely to DNA intermediates . For both types of element, recombination reactions involved in integration are carried out by element-specific enzymes . These are called transposases in the case of DNA elements and integrases in the case of the best-characterized RNA elements, the retroviruses and retrotransposons . In spite of major differences between these two transposition strategies, one step in the process, that of insertion, appears to be chemically identical . Current evidence suggests that the similarities in integration mechanism are reflected in amino acid sequence similarities between the integrases and many transposases . These similarities are particularly marked in a region which is thought to form part of the active site, namely the DDE motif . In the light of these relationships, we attempt here to compare mechanistic aspects of retroviral integration with transposition of DNA elements and to summarize current understanding of the functional organization of integrases and transposases. World J Surg, 1995 Jan-Feb, 19(1), 144 - 8; discussion 148-9 Fiber: effect on bacterial translocation and intestinal mucin content; Frankel W et al.; Total parenteral nutrition (TPN) and elemental diet (ED) produce intestinal atrophy and increase bacterial translocation (BT) to mesenteric lymph nodes . The increased rate of BT may be due to alterations in mucosal structure, enzyme activity, or mucin content . Fiber improves intestinal structure and function in rats and may reduce the rate of BT . This study determined whether the addition of fiber to TPN or ED would maintain intestinal integrity and decrease BT to the mesenteric lymph nodes . Fifty-six adult male Sprague-Dawley rats underwent placement of jugular catheters and were assigned to one of five dietary groups: TPN, TPN+oral oat fiber (TPNF) 2 g/day, ED, ED+oral oat fiber (EDF) 2 g/day, or AIN-76 (control); they were pair-fed for 7 days . On day 8 the mesenteric lymph nodes were removed for bacterial cultures; and jejunal mucosal weight, DNA, protein, alkaline phosphatase, maltase, and jejunal mucin content were measured . Enteral nutrition significantly decreased BT when compared to parenteral feeding, and fiber significantly decreased BT when administered to rats receiving TPN or ED . Improvements in intestinal mucosal structure were not consistently associated with decreased rates of BT . Additionally, BT occurred independently of jejunal mucin concentration . Mechanisms other than maintenance of mucosal structure or mucin content are important in the mediation of fiber-induced decreased BT in rats receiving TPN or ED. Electrophoresis, 1995 Jan, 16(1), 1 - 7 Pulsed field separation of large supercoiled and open-circular DNAs and its application to bacterial artificial chromosome cloning; Wang M et al.; We have studied the separation of large (80-300 kbp) supercoiled (SC) DNA in conventional agarose gel electrophoresis, field inversion gel electrophoresis (FIGE) and pulsed field gel electrophoresis (PFGE) . DNA migration was measured under a variety of electrophoretic conditions including different switch times, temperatures, agarose concentrations, and voltage gradients . The migration of SC DNA was found to be inversely proportional to its molecular weight in the three electrophoresis systems tested . In conventional agarose electrophoresis, voltage gradient was found to be the determining parameter in the separation of SC DNA . Unlike large linear DNAs, the migration of SC DNA was found to be independent of switch time in PFGE and FIGE . Broad DNA bands were observed in prolonged FIGE runs . In addition, we have also studied the migration of open-circular (OC) DNA (80 and 100 kbp) in pulsed field gel electrophoresis . Eighty kbp OC DNA can migrate into agarose gels under certain pulsed field conditions whereas 100 kbp OC DNA was trapped at the wells . Based on electrophoretic conditions described in this report, we can determine the size of bacterial artificial chromosome (BAC) clones without restriction enzyme digestion and have enriched the percentage of larger size clones in BAC cloning. Acta Pharm Hung, 1995 Jan, 65(1), 5 - 8 {Interaction between immunomodulatory drugs in mice with special regard to changes of drug sensitivity and rate of bacterial translocation}; Anderlik P et al.; In acute toxicity experiments the changes in drug sensitivity and in the rate of bacterial translocation (BT) were investigated in mice treated with immunomodulatory drugs: dianhydrogalactitol (DAG) in doses 20 and 30 mg/kg, chlorpromazine (CPZ) in doses 60 and 75 mg/kg and Mannozym (M) in dose 40 mg/kg for zymosan content . The drugs were used separately or in combination . The sensitivity of mice to immunosuppressive DAG or CPZ was higher in the case of combined treatment, than that of separately treated ones . The rate of BT was also higher in combined treated mice . The pretreatment with M that has immunostimulatory effect, influenced neither the sensitivity of mice to DAG or CPZ, nor the normal very low rate of BT . The present results reinforced the authors' earlier observations, that the effects of immunosuppressive drugs could cumulate and cause more serious damage of the organism . The authors suggest that the increase in drug sensitivity to immunosuppressive agents is in connection with increased rate of BT and effect of endotoxin. J Burn Care Rehabil, 1995 Jan-Feb, 16(1), 27 - 30 An experimental study to determine the effects of Dermagraft on skin graft viability in the presence of bacterial wound contamination; Economou TP et al.; Dermagrafts (Advanced Tissue Science, La Jolla, Calif.) is a possible dermal substitute currently in early stages of clinical trials . It consists of polyglycolic acid mesh impregnated with viable, human, neonatal fibroblasts . The randomized prospective study with the mouse model was undertaken to determine the effect of Dermagraft on skin graft viability in the presence of wound contamination with controlled concentrations of commonly encountered burn wound pathogens . Appropriate controlled series were run concurrently . Placement of Dermagraft, or polyglycolic acid mesh, had no significant effect on skin graft viability when compared with simple skin grafts . Controlled bacterial contamination of skin grafts with Dermagraft did not significantly change the occurrence of graft viability when compared with control groups of skin grafts with controlled bacterial contamination . These studies suggest that Dermagraft does not increase the occurrence of graft loss in the face of wound bed contamination. Biophys J, 1995 Jan, 68(1), 360 - 3 Observation of the helical structure of the bacterial polysaccharide acetan by atomic force microscopy; Kirby AR et al.; A method has been developed that has been found to give reproducible images of uncoated polysaccharides by Atomic Force Microscopy (AFM) . Aqueous solutions of the polysaccharide are deposited as drops onto freshly cleaved mica surfaces, air dried, and then imaged under butanol . The method has been used to obtain images of the bacterial polysaccharide acetan . In regions within the deposited sample, where the molecules are aligned side-by-side, it has been possible to observe a periodic structure along the polysaccharide chain, attributable to the helical structure of acetan. Int J Technol Assess Health Care, 1995 Winter, 11(1), 11 - 25 A cost-outcome description of the septic work-up for bacterial infection in neonates in a tertiary care hospital; Thomas RE et al.; Analysis of the septic work-up of 194 neonates at Women's College Hospital, Toronto, showed that the only antepartum condition predicting neonatal sepsis was the mother being on antibiotics . The only postnatal condition predicting sepsis was a maternal postpartum white blood cell count over 11,000 . The average cost for tests for a septic work-up in these 194 mother-neonate pairs was $71.48 (Canadian dollars), and the average cost of tests to find a septic case was $1,066.77. Arch Tierernahr, 1995, 47(3), 287 - 94 Effect of methionine and its related compounds on rumen bacterial activity; Hegedus M et al.; The effect of several methionine sources (L-methionine = L-MET; DL-methionine = DL-MET, DL-S-methyl-methionine-sulphonium-chloride = SMM; N-hydroxymethyl-DL-methionine-Ca = NHM; methionine-hydroxy-analog free acid = MHA; methionine-sulphoxide = MSO) on rumen bacterial growth was studied using a new methodical approach which utilises a methionine free assay medium (Bacto Methionine Assay Media, Difco) supplemented by increasing quantities of the methionine sources and inoculated with one drop of diluted rumen bacteria . The optical density was measured after 18 h incubation on 39 degrees C . L- and DL-MET promoted the highest growth response, while SMM and NHM exerted significantly (p < 0.05) lower optical densities . MHA and MSO showed no growth response . The methodical approach and the possible bacterial strains, which might have contributed to the growth response have been discussed. Vojnosanit Pregl, 1995 Jan-Feb, 52(1), 49 - 57 {Contribution of fiber optic bronchoscopy to the etiologic diagnosis of bacterial pneumonia}; Ponomarev D et al.; In 24 patients with bacterial pneumonia, reliability of the samples routinely taken for etiologic diagnosis (sputum, throat swab, bronchial brushing, bronchoalveolar lavage fluid--BALF, blood, pleural fluid) was determined . Organisms detected in blood, pleural fluid, transbronchial biopsy (TBB) or percutaneous transthoracic needle aspiration biopsy (PTNAB) samples were considered as truly causative, whereas those isolated in at least two various samples from a single patient were considered as presumably causative . Most sensitive diagnostic samples were BALF, TBB and PTNAB (100% each) . However, the specificity of BALF was very low (17%) . Bronchial aspirate was highly sensitive (95%) but not specific (14%) . Bronchial brushing was sensitive (86%) but its specificity low (14%) . Sputum was hardly sensitive (40%) and had no specificity . Throat swab had virtually no diagnostic value because of its low sensitivity (10.5%) and specificity (50%). J Comp Physiol {B}, 1995, 165(2), 85 - 92 Bacterial lipopolysaccharide (LPS) and interleukin 1 (IL-1) exert multiple physiological effects in the tilapia Oreochromis mossambicus (Teleostei); Balm PH et al.; To gain insight in immuno-endocrine communication in teleosts the physiological effects of interleukin 1 and bacterial lipopolysaccharide in teleosts were investigated . Tilapia (Oreochromis mossambicus) were treated with murine interleukin 1 and E . coli lipopolysaccharide in vivo, and lipopolysaccharide was administered to pituitary lobes and head kidneys in vitro . The integument of the fish appeared to be a sensitive target for the preparations tested, since proliferation of chloride cells and of epidermal mucous cells was observed as well as an increase in epidermal thickness . These effects may relate to an acute phase-like reaction caused by the treatments . Lipopolysaccharide administration furthermore resulted in an increase in plasma free fatty acids levels . Lipopolysaccharide, but not interleukin 1, stimulated the interrenal axis of the fish, as judged by the increase in cortisol production measured in superfusion of head kidneys . In addition to these in vivo effects, lipopolysaccharide also displayed several effects in vitro . Pituitary adrenocorticotropic hormone, as well as alpha-melanocyte stimulating hormone, release was inhibited, and the head kidney responsiveness to adrenocorticotropic hormone was inhibited after pretreatment of the tissue with the E . coli product . This latter effect coincided with the release of an unidentified alpha-melanocyte stimulating hormone immunoreactive fraction by the head kidneys which could be stimulated by lipopolysaccharide . The data strongly support the notion that the immune system is involved in adaptive regulations in teleosts, and that immunoendocrine interactions are phylogenetically old mechanisms. Probl Tuberk, 1995, (3), 2 - 4 {Organization of epidemiological measures in surroundings of patients with active pulmonary tuberculosis with undetermined bacterial discharge}; Samusevich LN; It is stated that family members and neighbours of patients with active tuberculosis of the lungs with indefinite bacterial discharge are at higher risk to develop tuberculosis than other population . In such foci of tuberculosis prompt antituberculosis measures are recommended even though final results on bacterial discharge of the patient are not yet available. Can J Microbiol, 1995, 41 Suppl 1, 94 - 105 Considerations on the structure and biochemistry of bacterial polyhydroxyalkanoic acid inclusions; Steinbuchel A et al.; Some mathematical calculations were done that provided information about the structure and biochemistry of polyhydroxyalkanoic acid (PHA) granules and about the amounts of the different constituents that contribute to the PHA granules . The data obtained from these calculations are compared with data from the literature, which show that PHA granules consist not only of the polyester but also of phospholipids and proteins . The latter are referred to as granule-associated proteins, and they are always located at the surface of the PHA granules . A concept is proposed that distinguishes four classes of structurally and functionally different granule-associated proteins: (i) class I comprises the PHA synthases, which catalyze the formation of ester linkages between the constituents; (ii) class II comprises the PHA depolymerases, which are responsible for the intracellular degradation of PHA, (iii) class III comprises a new type of protein, which is referred to as phasins and which has most probably a function analogous to that of oleosins in oilseed plants, and (iv) class IV comprises all other proteins, which have been found to be associated with the granules but do not belong to classes I-III . Particular emphasis is placed on the phasins, which constitute a significant fraction of the total cellular protein . Phasins are assumed to form a close protein layer at the surface of the granules, providing the interface between the hydrophilic cytoplasm and the much more hydrophobic core of the PHA inclusion. J Anim Sci, 1995 Jan, 73(1), 220 - 27 Studies on the in situ nitrogen degradability corrected for bacterial contamination of concentrate feeds in steers; Beckers Y et al.; Stable 15N was used to evaluate the influence of bacterial contamination on in situ DM and N degradabilities (Dg) of meat and bone meal (MBM), soybean meal (SBM), and wheat bran (WB) in two steers . Bacterial DM and N contamination ranged from 2.4 to 28.6% and 2.1 to 56.8% of residual DM and N, respectively . Effective N degradability increased when bacterial contamination was taken into account (P < .05) . The difference was low for MBM (2.4%) and for SBM (3.4%) but high for WB (12.2%) . Theoretically, using solid-associated bacteria should give the most accurate correction for bacterial contamination; however, results showed that Dg of N based on liquid-associated bacteria were identical for MBM and SBM (P > .05) and different for WB (P < .05) . In a second experiment, five treatments were applied to incubated feeds to remove bacteria fixed to the residues and consequently to determine directly the Dg of DM and N corrected for the bacterial contamination without the need for a marker . These treatments involved chilling for 6 h at 4 degrees C in saline solution alone (T1) or with a commercial detergent (T2), or with sodium dodecyl sulfate (T3) or with methylcellulose (T4), followed by pummeling in a stomacher for 5 min . The last treatment was only machine washing twice (T5) . The Dg of DM can be directly determined following the first four treatments, nevertheless their application to MBM and SBM led to higher Dg of N than that corrected for the bacterial contamination determined in the first experiment (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) Ann Trop Paediatr, 1995, 15(1), 55 - 9 Inflammatory response in bacterial meningitis: cytokine levels in the cerebrospinal fluid; Low PS et al.; Inflammatory response plays an important role in the pathogenesis of cerebral injury in bacterial meningitis . In this study, we evaluated the cytokine levels of interleukin 1-beta (IL1 beta), tumour necrosis factor alpha (TNF alpha) and interleukin 6 (IL6) in the cerebrospinal fluid (CSF), and determined their correlation with acute clinical complications and with changes in CSF biochemistry . Interleukin 6, TNF alpha and IL1 beta were present in 9/9, 3/9 and 4/9 patients, respectively . The CSFs with detectable TNF alpha or IL1 beta had higher levels of IL6 (p < 0.02), protein (NS) and lower glucose levels (p < 0.02), compared with those in which TNF alpha and IL1 beta were absent . Tumour necrosis factor alpha and IL1 beta levels also correlated with the presence of prolonged fever, fits, spasticity and death (logTNF alpha: r = 0.70, p < 0.05; logIL1 beta: r = 0.62, p = 0.08) . The cytokine levels reflect the degree of inflammatory response and are positively correlated with the severity of acute clinical complications . Modulation of this inflammatory response in bacterial meningitis may improve its morbidity and mortality. Pneumonol Alergol Pol, 1995, 63(5-6), 253 - 8 {Influence of bacterial infection and formalin vapors on the course of ovalbumin induced experimental bronchospastic reaction in guinea pigs}; Pindel B et al.; The pathomechanism of the bronchospastic reaction is not fully explained . Ovalbumin induced bronchospastic reaction in guinea pigs is widely accepted as a classical experimental model and was appleid in this study . The bronchoconstriction, bronchial hypersensitivity and humoral immune response were measured after bronchial infection and chemical injury by formalin vapours . The intensity of the bronchospasm was measured by Lundberg index, the haemolytic activity of complement and the level of circulating immune complexes were measured at the beginning and at the end of the experiment . The increase of the bronchospastic reaction and bronchial hypersensitivity was observed bacterial infection and after formalin vapours too . Bronchial infection and chemical irritation of bronchial tree lead to the increase of the circulating immune complexes level and to the decrease of the haemolytic activity of the complement. Annu Rev Biochem, 1995, 64, 141 - 69 DNA processing reactions in bacterial conjugation; Lanka E et al.; Bacterial conjugation is an important source of genetic plasticity . The initiation complex for conjugative transfer of transmissible plasmids--the relaxosome--is a specific DNA-protein structure that has been isolated from cells and reconstituted from purified components in vitro . Complexes containing uncleaved DNA and DNA cleaved at the nicsite in the origin of transfer (oriT) coexist in equilibrium . Relaxase is usually loaded onto oriT by accessory DNA-binding proteins . Relaxase catalyzes cleavage of a specific phosphodiester bond at nic and becomes covalently linked through a tyrosyl residue to the 5' terminus of the cleaved strand . Cleaved DNA may be unwound for transfer by a plasmid-encoded helicase . Single-strand transfer is thought to occur by a replicative rolling circle mechanism . Termination of a round of transfer is achieved by the cleaving-joining activity of the relaxase linked to the 5' end of the transferring strand . Relationships between DNA processing reactions and conjugative interactions of cell envelopes are particularly obscure aspects of the conjugation cycle. Biochem Mol Biol Int, 1995 Jan, 35(1), 177 - 83 Pentamethyl-hydroxychromane, vitamin E derivative, inhibits induction of nitric oxide synthase by bacterial lipopolysaccharide; Hattori S et al.; Vitamin E, a lipophilic antioxidant, has effectively inhibited the activation of cytokine-induced nuclear factor kB (NFkB) . Since NFkB plays a critical role in the induction of an isoform of nitric oxide synthase (iNOS) gene by lipopolysaccharide (LPS), we investigated the effect of a vitamin E derivative, pentamethyl-hydroxychromane (PMC), which is an extremely potent inhibitor of NFkB activation, on the induction of nitric oxide (NO) synthesis and iNOS mRNA by LPS . PMC inhibited the LPS-stimulated induction of NO production in a concentration-dependent fashion in cultured J774 macrophages and rat vascular smooth muscle cells without evidence of cytotoxicity . However, the addition of PMC to J774 macrophages after the induction of iNOS did not inhibit NO production . Treatment of J774 macrophages with LPS resulted in a significant expression of iNOS mRNA, which was profoundly reduced by PMC . Data suggest that PMC inhibits the induction of iNOS by preventing iNOS gene expression through inhibition of NFkB activation. Mutat Res, 1995 Jan, 341(3), 185 - 92 The gradient plate assay: a modified Ames assay used as a prescreen for the identification of bacterial mutagens; Rexroat MA et al.; Bacterial test systems have been used extensively to identify the mutagenic potential of new compounds . In particular, the Ames test has gained worldwide acceptance and is required by many regulatory agencies to support product registration . The gradient plate assay (GPA) is a modification of the Ames test . It is used as a high capacity prescreen to detect the mutagenic potential of synthetic intermediates, impurities, and research compounds over a concentration gradient . Since the development of the GPA, over 4000 compounds have been tested in the assay . Selection and use of the GPA in our laboratory is due to many factors: reliability; sensitivity; capacity; timeliness of reporting results; and establishment of safety standard in the laboratory . In this manuscript, results of the GPA method are compared with results from the traditional Ames assay . To date, 113 compounds of identical lots have been evaluated in both tests, and in all but 3 instances the results are the same . Thus, the GPA is an ideal assay for use as a prescreen in determining the ability of a compound to induce bacterial mutation. Infect Immun, 1995 Jan, 63(1), 51 - 6 CD14 and tissue factor expression by bacterial lipopolysaccharide-stimulated bovine alveolar macrophages in vitro; Yang Z et al.; The membrane-associated CD14 receptor (mCD14) is a monocyte/macrophage differentiation antigen, and it has been demonstrated to serve as a receptor for bacterial lipopolysaccharide (LPS; endotoxin) . Binding of LPS to mCD14 has been shown to be associated with LPS-induced macrophage, monocyte, and neutrophil activation in humans . In this report, we describe the presence and function of an mCD14-like receptor on bovine alveolar macrophages (bAM) . An immunofluorescence technique and flow cytometric analysis indicated binding of anti-human CD14 monoclonal antibodies (MAb) My4, 3C10, and 60bd to bAM . Binding of anti-CD14 MAb (3C10 and MY4) was reduced over 20% by pretreatment of bAM with phosphatidylinositol-specific phospholipase C (0.5 to 1.0 U/ml), indicating that bovine mCD14 is a glycosyl phosphatidylinositol-anchored protein . In addition, pretreatment of bAM with anti-CD14 MAb decreased binding of 125I-labeled LPS to macrophages, suggesting that bovine mCD14 serves as a receptor for LPS . A cDNA probe based on the human sequence for CD14 was used in Northern (RNA) blot analysis, and hybridization to human monocyte CD14 yielded the expected 1.5-kb band . Hybridization to bovine mRNA yielded a 1.5-kb band plus an unexpected 3.1-kb band . Constitutive expression of bovine CD14 mRNA was observed, and the expression level was modestly elevated in bAM stimulated for 24 h with LPS (1 ng/ml) in the presence of bovine serum . The function and activation of bAM were assessed by quantitation of tissue factor (TF) expression on the cells using an activated factor X-related chromogenic assay and S-2222 substrate . LPS (1 ng/ml)-mediated upregulation of TF expression on bAM was dependent on the presence of bovine serum components, and TF expression was inhibited by anti-CD14 MAb . In addition, TF mRNA levels in LPS-stimulated bAM were decreased by pretreatment of cells with anti-CD14 MAb (MAb 60bd, 10 micrograms/ml). Infect Immun, 1995 Jan, 63(1), 104 - 9 Cytokines and progenitor cells of granulocytopoiesis in peripheral blood of patients with bacterial infections; Selig C et al.; To investigate the physiological role of granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in the adaptation mechanisms of myelopoiesis to enhanced demand, we studied both cytokines and their myeloid target cells in hematologically healthy patients suffering from acute bacterial infections . Endogenous serum levels of G-CSF and GM-CSF, granulocyte-macrophage colony-forming cell (GM-CFC) concentrations, and differential counts were determined for the peripheral blood of 57 patients with clinically apparent bacterial infections (26 males and 31 females aged 16 to 89 years) and 18 healthy controls (8 males and 10 females aged 23 to 84 years) . Patients were selected for acute-phase protein and at least two additional clinical signs reflecting a bacterial infection . Patients showed significantly higher numbers of myeloid progenitor cells than controls (median, 68 versus 26 GM-CFC/ml; P < or = 0.01) . G-CSF but not GM-CSF levels were found to be elevated (> or = 50 to 863 pg/ml) . In the acute stage of infection, progenitor and cytokine levels were not influenced by gender, differences in therapy, or localization of the infection . Progenitor and G-CSF levels were not associated with absolute neutrophil counts or C-reactive protein . However, a negative correlation between number of GM-CFC per milliliter and age (R = -0.47; P < or = 0.001) and an inverse relationship between the incidence of high GM-CFC concentrations and elevated G-CSF levels (phi = -0.34; P < or = 0.01) were found . Combining both parameters into a cytokine-progenitor pattern, we observed a highly significant age-dependent response of myelopoiesis to inflammation (P < or = 0.001) . Younger patients had high progenitor counts (> 75 GM-CFC/ml) associated with G-CSF levels below 50 pg/ml, whereas for the older patients, the reverse pattern was predominant . The results indicate that the age-dependent myelopoietic response to acute bacterial infections is characterized by an inverse relationship between progenitor cells and G-CSF . The observed cytokine-progenitor patterns could have implications for therapy with G-CSF and the prognosis of infectious diseases. Biochem Biophys Res Commun, 1994 Dec 30, 205(3), 1644 - 50 Novel bacterial P-type ATPases with histidine-rich heavy-metal-associated sequences; Trenor C 3rd et al.; Menkes disease and Wilson disease are human disorders of copper metabolism . It has recently been shown that both are due to mutations in P-type ATPase copper transport molecules . Related heavy metal transporting ATPases have been described in several strains of bacteria . In an effort to isolate other mammalian metal transporters, we screened a human small intestine library with probes homologous to conserved sequences in the known proteins . Two novel cDNAs were isolated, which encode new members of this family . Surprisingly, they were both of bacterial origin, most likely derived from E . coli sequences transduced during library construction. Proc Natl Acad Sci U S A, 1994 Dec 20, 91(26), 12701 - 5 Coherent nuclear dynamics at room temperature in bacterial reaction centers; Vos MH et al.; A room-temperature study is reported of the femtosecond spectral evolution of the stimulated emission band of the primary electron-transfer precursor P* in bacterial photosynthesis . The study was performed with membranes of the antenna-deficient RCO1 mutant of Rhodobacter sphaeroides . A time-dependent red shift, reflecting nuclear motion out of the Franck-Condon region of the excited state, is resolved . Analysis of oscillatory features persisting for > 1 ps in the kinetics revealed main frequencies of the activated motions at 30, 84, 145, and 192 cm-1 . The oscillations occur on the time scale of primary electron transfer . Our results set a lower limit for the vibrational dephasing time in P* that is not compatible with the usual assumption in theoretical treatments of complete vibrational relaxation prior to electron transfer, even at room temperature. Biochemistry, 1994 Dec 6, 33(48), 14378 - 86 Binding sites of quinones in photosynthetic bacterial reaction centers investigated by light-induced FTIR difference spectroscopy: assignment of the interactions of each carbonyl of QA in Rhodobacter sphaeroides using site-specific 13C-labeled ubiquinone; Breton J et al.; Light-induced QA-/QA FTIR difference spectra of the photoreduction of the primary quinone (QA) have been obtained for Rhodobacter sphaeroides reaction centers (RCs) reconstituted with ubiquinone (Q3) labeled selectively with 13C at the 1- or 4-position of the quinone ring, i.e., on either of the two carbonyls . The vibrational modes of the quinone in the QA site are compared to those in vitro . IR absorption spectra of films of the labeled quinones show that the two carbonyls contribute equally to the split C = O band at 1663-1650 cm-1 . This splitting is assigned to the two different geometries of the methoxy group nearest to each carbonyl . The QA-/QA spectra of RCs reconstituted with either 13C1- or 13C4-labeled Q3 and with unlabeled Q3 as well as the double differences calculated from these spectra exhibit distinct isotopic shifts for the bands assigned to C = O and C = C vibrations of the neutral QA . For the unlabeled QA, these bands correspond to the bands at 1660, 1628, and 1601 cm-1 previously detected upon nonselective isotopic labeling {Breton, J., Burie, J.-R., Berthomieu, C., Berger, G., & Nabedryk, E . (1994) Biochemistry 33, 4953-4965} . The 1660-cm-1 band is unaffected upon selective labeling at C4 but shifts to approximately 1623 cm-1 upon 13C1 labeling, demonstrating that this band arises from the C1 carbonyl, proximal to the isoprenoid chain . The band at 1628 cm-1 shifts by 11 and 16 cm-1 upon 13C1 and 13C4 labeling, respectively, and is assigned to a C = C mode coupled to both carbonyls.(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1994 Dec 2, 269(48), 30206 - 11 Probing protein-protein interactions . The ribose-binding protein in bacterial transport and chemotaxis; Bjorkman AJ et al.; A number of mutations at Gly134 of the periplasmic ribose-binding protein of Escherichia coli were examined by a combined biochemical and structural approach . Different mutations gave rise to different patterns of effects on the chemotaxis and transport functions . The smallest residue (alanine) had the least effect on transport, whereas large hydrophobic residues had the smallest effect on chemotaxis . Comparison of the x-ray crystal structure of the G134R mutant protein (2.5-A resolution) to that of the wild type (1.6-A resolution) showed that the basic structure of the protein was unaltered . The loss of chemotaxis and transport functions in this and similar mutant proteins must therefore be caused by relatively simple surface effects, which include a change in local main chain conformation . The loss of chemotaxis and transport functions resulting from the introduction of an alanine residue at position 134 was suppressed by an additional isoleucine to threonine mutation at residue 132 . An x-ray structure of the I132T/G134A double mutant protein (2.2-A resolution) showed that the changes in local structure were accompanied by a diffuse pattern of structural changes in the surrounding region, implying that the suppression derives from a combination of sources. Gene, 1994 Dec 2, 150(1), 187 - 92 Expression of the pheromone 3-encoding gene of Euplotes octocarinatus using a novel bacterial secretion vector; Brunen-Nieweler C et al.; The pheromone 3-encoding gene (phr3) of Euplotes octocarinatus was expressed in Escherichia coli using a novel expression-secretion vector . The vector, pExSec1, contains a strong and tightly regulated T7 promoter, the corresponding Shine-Dalgarno sequence and the T7 terminator region . Translation starts at the protein A leader sequence followed by the synthetic ZZ sequence of protein A . The expression-secretion modules are embedded in the high-copy-number plasmid vector, pUK21, which carries a kanamycin-resistance marker (KmR) . The produced ZZ-pheromone 3 (Phr3) fusion protein was secreted into the culture medium of the host cells . It was isolated by affinity chromatography and was further purified by gel filtration . After refolding with protein disulfide isomerase (PDI), the fusion protein exhibited the same high activity as the native pheromone. J Surg Res, 1994 Dec, 57(6), 682 - 6 The role of the mucus gel layer in intestinal bacterial translocation; Maxson RT et al.; The mucus gel layer is thought to be a vital component of the intestinal mucosal barrier . The purpose of this study is to determine if decreasing mucus production following ischemia and reoxygenation of the intestinal mucosa would alter bacterial translocation in an in vitro rat mucosal model . Clonidine was used to decrease mucus production associated with an ischemia/reoxygenation insult . Bacterial translocation was studied in a modified Ussing chamber using Escherichia coli K100 . The quantity of mucus produced, as well as the incidence and quantity of bacteria translocating was measured . In the clonidine-treated animals, there was a significant decrease in the amount of mucus produced compared to the control animals . The clonidine animals also had a higher quantity of bacteria translocating during the reoxygenation period compared to the control animals . The mucus gel layer is protective against in vitro translocation of bacteria following an ischemia/reoxygenation insult. Ann Surg, 1994 Dec, 220(6), 798 - 808 Secretory immunoglobulin A, intestinal mucin, and mucosal permeability in nutritionally induced bacterial translocation in rats; Spaeth G et al.; OBJECTIVE . The authors investigated the role of mucin and secretory immunoglobulin A (slgA) in a model of nutritionally induced bacterial translocation . BACKGROUND . Parenteral and certain elemental diets have been shown to impair intestinal barrier function, whereas fiber has been shown to protect against nutritionally induced bacterial translocation . However, the factors responsible for these phenomenon have not been fully determined . METHODS . Intestinal mucin levels, mucosal protein content, slgA, intestinal morphology, and permeability to horseradish peroxidase, bacterial translocation, and intestinal bacterial population levels were measured in rats 7 days after receiving total parenteral nutrition (TPN) solution (28% glucose, 4.25% amino acids; 307 kcal/kg/day) enterally (ORAL-TPN) or parenterally (IV-TPN) with or without enteral bulk fiber supplementation . Chow-fed rats served as control subjects . RESULTS . The incidence of bacterial translocation in the ORAL-TPN and IV-TPN groups was reduced significantly by the provision of fiber (p < 0.05) . Mucosal protein, slgA, and insoluble mucin levels were decreased in the jejunum of the ORAL-TPN and IV-TPN groups, with mucosal protein levels being decreased to a greater extent than slgA or mucin . Although similar decreases in these parameters were observed in the fiber-fed groups, fiber appeared to improve intestinal barrier function as measured by horseradish peroxidase permeability . CONCLUSIONS . The provision of bulk-forming fiber improves intestinal barrier function as measured by peroxidase permeability and bacterial translocation, but does not restore mucosal protein content, intestinal mucin, or slgA levels to normal. Hepatology, 1994 Dec, 20(6), 1495 - 501 Renal impairment after spontaneous bacterial peritonitis in cirrhosis: incidence, clinical course, predictive factors and prognosis; Follo A et al.; Although spontaneous bacterial peritonitis is considered a precipitating factor of renal impairment in cirrhosis, no study specifically addressing this problem has been reported . This study was aimed at assessing the incidence, clinical course, predictive factors and prognosis of renal impairment in cirrhotic patients with peritonitis . Therefore, 252 consecutive episodes of spontaneous bacterial peritonitis in 197 patients were analyzed . Clinical and laboratory data obtained before and after diagnosis of peritonitis were considered as possible predictors of renal impairment and hospital mortality . Renal impairment occurred in 83 (33%) episodes, and in every instance it fulfilled the criteria of functional kidney failure . Renal impairment was progressive in 35 episodes, steady in 27 and transient in 21 . Blood urea nitrogen and serum sodium concentration before peritonitis and band neutrophils count in blood at diagnosis were independent predictors for the development of renal impairment . Renal impairment was the strongest independent predictor of mortality during hospitalization . Other independent prognostic factors were blood urea nitrogen level before peritonitis, age, positive ascitic fluid culture and serum bilirubin level during infection . These results indicate that renal impairment is a frequent event in cirrhotic patients with spontaneous bacterial peritonitis that occurs mainly in patients with kidney failure before infection . Renal impairment is the most important predictor of hospital mortality in cirrhotic patients with spontaneous bacterial peritonitis. Infect Immun, 1994 Dec, 62(12), 5689 - 93 Bacterial heat shock proteins directly induce cytokine mRNA and interleukin-1 secretion in macrophage cultures; Retzlaff C et al.; Bacterial heat shock proteins (hsp) have been shown to be important immunogens stimulating both T cells and B cells . However, little is known concerning the direct interactions between hsp and macrophages . In this study, we demonstrated that treatment of macrophage cultures with purified bacterial hsp, including Legionella pneumophila hsp60, Escherichia coli GroEL, Mycobacterium tuberculosis hsp70, Mycobacterium leprae hsp65, and Mycobacterium bovis BCG hsp65, increased the steady-state levels of cytokine mRNA for interleukin-1 alpha (IL-1 alpha), IL-1 beta, IL-6, tumor necrosis factor alpha, and granulocyte-macrophage colony-stimulating factor as well as supernatant IL-1 secretion . This effect was shown not to be due to contamination of the hsp preparations with bacterial lipopolysaccharide . However, not all hsp induced cytokines; M . tuberculosis hsp10 showed minimal activity in our study . These results suggest that bacterial hsp might modulate immunity by rapidly and directly increasing cytokine production in macrophages. Am J Respir Crit Care Med, 1994 Dec, 150(6 Pt 1), 1493 - 8 Drug smoking, Pneumocystis carinii pneumonia, and immunosuppression increase risk of bacterial pneumonia in human immunodeficiency virus-seropositive injection drug users; Caiaffa WT et al.; To examine the risk factors for the first episode of bacterial pneumonia among human immunodeficiency virus (HIV)-seropositive injection drug users (IDUs), medical record review was performed on IDUs participating in a cohort study from January 1988 to June 30, 1992 . HIV-seropositive IDUs with a first episode of bacterial pneumonia (n = 40) were matched with up to five HIV-seropositive control subjects without bacterial pneumonia (n = 197) by date of entry (+/- 3 mo) and length of follow-up . Odds ratios (OR) were estimated using conditional logistic regression . The incidence of bacterial pneumonia was 1.93 in 100 person-years in HIV seropositive and 0.45 in 100 person-years in HIV seronegative subjects (relative incidence = 4.3; 95% CI 2.4 to 7.5) . In univariate analyses, CD4 lymphocyte count < 200 cells/microliters previous episode of Pneumocystis carinii pneumonia (PCP), age between 30 and 40 yr and smoking illicit drugs (marijuana, cocaine, or crack) were associated with bacterial pneumonia . Cigarette smoking was associated with an increased odds of bacterial pneumonia (OR = 2.0), but this was not statistically significant because it was nearly universal in this cohort . In multivariate analysis, CD4 < 200 cells/microliters (OR = 6.75, 95% CI 2.13 to 21.42) and smoking illicit drugs (OR = 2.24, 95% CI 1.03 to 4.89) remained significantly associated with bacterial pneumonia . The odds ratio for cigarette smoking in the final model remained at 2.08 but was still not significant (95% CI 0.49 to 8.70) . Smoking illicit drugs had the strongest effect on risk of bacterial pneumonia among HIV-seropositive IDUs with a previous history of PCP (OR = 22.94; 95% CI 2.18 to 241.10). Int J Biochem, 1994 Dec, 26(12), 1391 - 3 Two nuclear-coded subunits of mitochondrial complex I are similar to different domains of a bacterial formate hydrogenlyase subunit; Videira A et al.; A computer comparison of protein sequences revealed similarity between the 30.4 kDa subunit of complex I from the fungus Neurospora crassa and the ORF5 subunit of formate hydrogenlyase from Escherichia coli . The ORF5 protein was previously known to be homologous to the 49 kDa component of the mitochondrial enzyme . We show that the 30.4 kDa corresponds to the N-terminal part while the 49 kDa subunit corresponds to the C-terminal portion of the bacterial protein . Thus, this bacterial protein represents a fusion of the two mitochondrial polypeptides suggesting that the two complex I genes arose from a single ancestor . Our results indicate that the 30.4 kDa and 49 kDa subunits are part of a structural and functional unit in complex I. Minerva Ginecol, 1994 Dec, 46(12), 657 - 61 {Bacterial vaginosis . Prevention of recurrence}; Ceruti M et al.; Bacterial vaginosis (BV) is the main cause of vaginitis . The condition is characterised by an abundant and odorous vaginal loss, but more than half the patients with demonstrable signs of BV do not report symptoms at all . Gardnerella vaginalis (Gv) is often associated with BV, but it is not the sole factor responsible, as is shown by the fact that it can be isolated in the vagina of women withBV . In 1992 and 1993, 2630 patients, 1460 of them gynaecological and 1170 obstetric, were admitted to the Obstetrics and Gynaecology Clinic of Parma University . Amsel criteria were adopted for diagnosing BV . Cases of BV were treated with 5 mg/die 2% clindamycin vaginal cream for 7 days . In the event of recurrences, 250 mg tablets of metronidazol were added: 8 tablets in 4 administrations in a single day, treatment also being extended to the partner . Patients admitted in 1993 received a protocol of hygienic and behavioural standards, stress being laid on prophylaxisa measures even after the end of therapy . BV proved to be present in 12.3% of cases, of whom only half were symptomatic . The situation was practically stationary if the 2 years are considered separately . Recurrences of symptomatic bacterial vaginosis were 15% in the absence of protocol application and 8.3% after the protocol . Recurrences were less frequent in the asymptomatic forms . Compared to the total number od cases of BV, recurrences were significantly low (12.1% p < 0.001). Photochem Photobiol, 1994 Dec, 60(6), 521 - 35 Protein structure modelling of the bacterial light-harvesting complex; Olsen JD et al.; Protein structure modelling offers a method of obtaining 3-dimensional information that can be tested and used to plan mutagenesis experiments when a crystallographically determined structure is not available . At its simplest a model may consist of little more than a secondary structure prediction coupled with a determination of the likely regions of transmembrane/membrane surface/globular configuration . These methods can yield an interesting topology map of the protein, which places the residues in their likely positions with respect to, for example, the membrane interface . If it is a member of a large family of related proteins then aligned protein sequences can be used to predict the residues that have an important function as these will be largely conserved in the alignments . Using all these methods a model can be constructed (using for example, the Nicholson Molecular Modelling Kit) to visualize the proposed structure in three dimensions following the premise of good design, that is, avoiding obvious steric clashes, packing of helices in a realistic manner, observing the correct H-bond lengths, etc . In this latter exercise the review of Chothia (Annu . Rev . Biochem . 53, 537-572, 1984) of the principles of protein structure is particularly helpful as it clearly sets out how proteins pack and their preferred configuration . There is a wealth of information about individual amino acid conformational preferences and observed frequencies of occurrence in known protein structures, which can help decide how the residues in the model can be oriented . In this article we have collated the various protein models of the bacterial light-harvesting complexes and present our own model, which is a synthesis of the available biophysical data and theoretical predictions, and show its performance in explaining recent results of site-directed mutants of the LH1 and LH2 light-harvesting complexes of Rhodobacter sphaeroides. Arzneimittelforschung, 1994 Dec, 44(12A), 1417 - 21 Protective effects of pidotimod against experimental bacterial infections in mice; Coppi G et al.; Pidotimod ((R)-3-{(S)-(5-oxo-2-pyrrolidinyl) carbonyl}-thiazolidine-4-carboxylic acid, PGT/1A, CAS 121808-62-6) protected mice against experimental bacterial infections in different experimental models . In all tests the drug's effect was measured as protection from death . The activity of pidotimod was evident and statistically significant after 5 administrations before the bacterial challenges . Pidotimod was active against many bacterial species infections, its active dosages ranging from 0.01 to 100 mg/kg i.p . x 5 times . Pidotimod showed against some bacterial infections a protection similar or better than those of bestatin, N-acetylmuramyl-L-Ala-D-isoGlu-OH and tuftsin . It showed high protection against bacterial infections in cyclophosphamide-immunodepressed mice . Finally, pidotimod showed an additive or synergic activity in combination with beta-lactam antibiotics (cefotaxime, ampicillin) against bacterial infections in mice. Br J Surg, 1994 Dec, 81(12), 1796 - 8 Bacterial translocation during peroperative colonic lavage of the obstructed rat colon; Horgan AF et al.; Peroperative antegrade colonic lavage is often performed before primary anastomosis in emergency colonic surgery . The influence of colonic lavage on bacterial translocation from the obstructed colon was determined . Forty female Wistar rats were studied in four groups: (1) control; (2) non-obstructed with lavage; (3) obstructed; and (4) obstructed with lavage . Ligature obstruction of the rectum was performed in groups 3 and 4 . Some 4 days later 35S-radiolabelled Escherichia coli was inoculated into the colon of all animals . Groups 2 and 4 underwent colonic lavage . Lavage in the group 4 animals with left-sided colonic obstruction significantly increased the levels of E . coli in regional nodes, liver, spleen, lung, kidney and blood (as assessed by organ culture and scintillation counting) compared with those in groups 1, 2 and 3 (P < 0.05) . These results suggest that peroperative lavage of the obstructed colon significantly increases the level of bacterial translocation. Planta Med, 1994 Dec, 60(6), 541 - 5 Studies on the anti-tumour, anti-bacterial, and wound-healing properties of dragon's blood; Chen ZP et al.; Three in-vitro assays have been adopted to examine the cytotoxicity and anti-bacterial activity of the blood-red sap of Croton lechleri from Ecuador, and to examine its effect upon the proliferation of endothelial cells . The sap was found not to be cytotoxic . Several simple phenolic compounds and diterpenes showed a potent anti-bacterial activity . The sap has little effect upon the proliferation of endothelial cells, and no single active ingredient was identified . A mechanism for the wound-healing property of the sap has been proposed. Eur J Immunol, 1994 Dec, 24(12), 3161 - 9 Elongated peptides, not the predicted nonapeptide stimulate a major histocompatibility complex class I-restricted cytotoxic T lymphocyte clone with specificity for a bacterial heat shock protein; Schoel B et al.; The peptides recognized by an H-2Db-restricted CD8 cytotoxic T lymphocyte (CTL) clone which is specific for the 60-kDa mycobacterial heat shock protein (hsp) and cross-reacts with stressed host cells were characterized . None of the nonapeptides from hsp60 conforming to the H-2Db binding motif were able to sensitize target cells for lysis by this CTL clone . Sequence analysis of the stimulatory fraction from a trypsin digest of hsp60, together with synthetic peptide studies, defined a cluster of overlapping epitopes . Carboxy-terminal extension by at least one amino acid of the nonamer predicted to bind best to H-2Db was essential for CTL recognition . Two such elongated peptides, a 10-mer and a 12-mer stimulated the clone at similarly low concentrations in the 100 pM range . We assume that these two peptides comply best with the natural epitope . In contrast, the 11-mer was inactive . The stimulatory 10-mer bound to H-2Db with an efficacy similar to that of the nonapeptide corresponding to the H-2Db motif, as revealed by peptide induced major histocompatibility complex (MHC) surface expression on RMA-S cells and competitive blocking of epitope recognition by the nonamer . Binding of these carboxy-terminally extended peptides to the MHC groove can be explained by anchoring through the amino acid residue Asn in position 5 of the peptide and by intrusion of the hydrophobic carboxy-terminal Ala (10-mer) or Leu (12-mer), but not Gly (11-mer), into the hydrophobic pocket of the H-2Db cleft . Because the carboxy-terminal part is thus larger than predicted, this region of the peptide may arch up from the binding groove . We assume that recognition of steric components of the MHC/peptide complex broaden the range of epitope specificity for a single T cell receptor . This flexibility not only promotes recognition of several overlapping peptides from a single antigen, but may also increase the chance of cross-reaction with similar peptides from unrelated proteins, including autoantigens . Consistent with this latter assumption, the T cell clone cross-recognizes mycobacterial hsp60 and stressed host cells. Int J Biol Macromol, 1994 Dec, 16(6), 343 - 7 Formation, derivatization and applications of bacterial cellulose; Geyer U et al.; Acetobacter xylinum produces highly crystalline cellulose extracellularly using glucose as a carbon source . The polymer formed is free of other biogenic compounds, separable in a simple way and characterized by its high water-absorption capacity . Stepwise solvent exchange from water to unpolar solvents leads to a drastic decrease of the water content of the bacterial cellulose without decrease of the highly swollen and activated state . Heterogeneous as well as homogeneous derivatizations, e.g . carboxymethylation, silylation and acetylation, were performed on the wet or dried biopolymer . Furthermore, different methods for formation of hollow fibres during biosynthesis were investigated . Such tubes may have applications as biocompatible material in medicine. J Pharm Pharmacol, 1994 Dec, 46(12), 1000 - 3 Iron-binding affinity of bacterial vaccine polysaccharides which contain phosphodiester linkages as part of the polymer chain and of other polyphosphates, including DNA; Bingham BR et al.; The interaction of iron (II) with bacterial polysaccharides, possessing phosphodiester bonds as part of their polymer chain, has been studied by equilibrium binding dialysis using atomic absorption spectrophotometry . Ferrous ions were found to bind with a stoichiometry of one per two phosphates and with a binding constant of about 2.5 x 10(3) M-1 . Similar results, but with larger (ca 1 x 10(4) M-1) binding constants were observed with DNA . This interaction helps explain the depolymerization of polyphosphates which has been observed in the presence of iron salts, and highlights the need to avoid iron contamination of vaccines (and other substances) which contain phosphodiester bonds . The interaction may also be a means of iron sequestration in bacteria which possess these cell-surface polyphosphates. Curr Opin Struct Biol, 1994 Dec, 4(6), 814 - 22 Unraveling a bacterial hexose transport pathway; Herzberg O et al.; Structural information about proteins involved in bacterial hexose transport mediated by the phosphoenolpyruvate:sugar phosphotransferase system is rapidly accumulating . Within the past year, two crystal structures and two solution NMR structures of the histidine-containing phosphocarrier protein have been reported, adding structural details to previous NMR and crystallographic work on this protein and on enzyme IIA . The crystal structure of the regulatory complex between the glucose enzyme IIA and glycerol kinase has been determined, and the association of the histidine-containing phosphocarrier protein and either the glucose enzyme IIA or the mannitol enzyme IIA have been studied by NMR . Proposals concerning the mechanism of phosphoryl transfer and the protein-protein interactions involved may now be tested more rigorously using these data. Lupus, 1994 Dec, 3(6), 507 - 14 Long-lasting effects of bacterial lipopolysaccharide promote progression of lupus nephritis in NZB/W mice; Granholm NA et al.; Lupus prone NZB/W mice repeatedly exposed to bacterial lipopolysaccharide (LPS) develop enhanced polyclonal B cell activation and exacerbated nephritis by a mechanism that results in increased deposits of immunoreactants in kidneys without measurable impairment of mononuclear phagocyte function . In this paper, we investigate whether the referenced effects of LPS are reversible after withdrawal of LPS, or whether their persistence could contribute to progression of nephritis to chronicity . In NZB/W mice previously exposed to LPS, features of enhanced polyclonal B cell activation, more severe glomerulonephritis with tubulointerstitial involvement, increased deposits of immunoreactants in glomeruli, and altered protein excretion persisted 6 weeks after LPS was discontinued . Additionally, mononuclear phagocyte function, assessed through liver uptake of radiolabeled immune complexes, was found to be impaired . The results indicate that some of the effects of prior exposure to LPS may be partially reversible; however, they last long after LPS has been discontinued, and additional impairment of immune function develops together with permanent glomerular dysfunction, thereby contributing to progression of nephritis to chronicity. Pediatr Res, 1994 Dec, 36(6), 799 - 804 L-selectin is down-regulated in umbilical cord blood granulocytes and monocytes of newborn infants with acute bacterial infection; Buhrer C et al.; The leukocyte glycoprotein L-selectin mediates an early step in the recruitment of leukocytes to sites of inflammation . L-Selectin surface expression is rapidly down-regulated by inflammatory signals in vitro . In a prospective study, we found L-selectin expression on umbilical cord blood granulocytes and monocytes to be significantly decreased in newborn infants with acute bacterial infection compared with controls (p < 0.01) . A significantly reduced L-selectin expression of both granulocytes and monocytes was also found to be associated with an increased neutrophil immature/total ratio (p < 0.01) but not with other laboratory markers of neonatal sepsis . There was no apparent impact of prematurity, low birth weight, gestational hypertension, or gestational diabetes on L-selectin expression . Although the mode of delivery did not affect granulocyte L-selectin expression, umbilical cord blood monocytes showed an increased L-selectin expression after emergency cesarean delivery compared with samples obtained after elective cesarean or vaginal delivery (p < 0.01) . We conclude that acute systemic inflammation results in down-regulation of granulocyte and monocyte L-selectin expression in vivo similar to that observed in vitro. Appl Environ Microbiol, 1994 Dec, 60(12), 4580 - 3 Renibacterium salmoninarum, the causative agent of bacterial kidney disease in salmonid fish, detected by nested reverse transcription-PCR of 16S rRNA sequences; Magnusson HB et al.; An assay based on reverse transcription and nested PCR amplification of hypervariable regions within the 16S rRNA sequence was used to specifically detect Renibacterium salmoninarum, the slowly growing causative agent of bacterial kidney disease in salmonid fish . This assay detected 1 to 10 bacteria per sample and took 1 to 2 days to perform . The assay was used to detect R . salmoninarum in ovarian fluid obtained from naturally infected fish . The assay was unreliable when it was used to examine kidney tissue. J Mol Evol, 1994 Dec, 39(6), 631 - 43 Tracing the spread of fibronectin type III domains in bacterial glycohydrolases; Little E et al.; The evolutionary spread of 22 fibronectin type III (Fn3) sequences among a dozen bacterial enzymes has been traced by searching databases with the non-Fn3 parts of the enzyme sequences . Numerous homologues were found that lacked the Fn3 domains . In each case the related sequences were aligned, phylogenetic trees were constructed, and the occurrences of Fn3 units on the trees were noted . Comparison with phylogenetic trees prepared from the Fn3 segments themselves allowed inferences to be made about when the Fn3 units were shuffled into their present positions. Neuron, 1994 Dec, 13(6), 1345 - 57 Agonist selectivity of glutamate receptors is specified by two domains structurally related to bacterial amino acid-binding proteins; Stern-Bach Y et al.; By exchanging portions of the AMPA receptor subunit GluR3 and the kainate receptor subunit GluR6, we have identified two discontinuous segments of approximately 150 amino acid residues each that control the agonist pharmacology of these glutamate receptors . The first segment (S1) is adjacent and N-terminal to the putative transmembrane domain 1 (TM1), whereas the second segment (S2) is located between the putative TM3 and TM4 . Only the simultaneous exchange of S1 and S2 converts the pharmacological profile of the recipient to that of the donor subunit . The two segments identified in this study share sequence similarities with the ligand-binding site of several bacterial periplasmic amino acid-binding proteins . Based on the X-ray structure of these proteins, we propose a model for the glutamate-binding site of ionotropic glutamate receptors. J Biol Chem, 1994 Nov 25, 269(47), 30049 - 55 Human glycyl-tRNA synthetase . Wide divergence of primary structure from bacterial counterpart and species-specific aminoacylation; Shiba K et al.; Several class I and class II human tRNA synthetases are clearly related to their bacterial counterparts . We report here the cloning, cDNA sequence, deduced primary structure, and expression in bacteria of a class II human glycyl-tRNA synthetase . While the human sequence aligns well with a Bombyx mori and a Saccharomyces cerevisiae sequence for glycyl-tRNA synthetase, particularly in the region of the class II-defining sequence motifs, it diverges widely from that of the Escherichia coli enzyme . The divergence is so great that from the sequences alone we cannot conclude that the human and E . coli proteins are descended from homologous genes . Moreover, even though the human and E . coli class II alanyl-tRNA synthetases cross-acylate their respective tRNAs, aminoacylations by the recombinant human and E . coli glycyl-tRNA synthetases are restricted to their homologous tRNAs . The species-specific aminoacylations correlate with a nucleotide sequence difference at a location in the acceptor stem that is known to be critical for aminoacylations by the E . coli enzyme . Thus, glycyl-tRNA synthetase may have followed a path of historical development different in at least some respects from that of several other tRNA synthetases. J Biol Chem, 1994 Nov 25, 269(47), 29920 - 7 Deducing the organization of a transmembrane domain by disulfide cross-linking . The bacterial chemoreceptor Trg; Lee GF et al.; The transmembrane domain of chemoreceptor Trg from Escherichia coli contains four segments, two from each subunit of the homodimer . We used site-specific mutagenesis to introduce cysteines into those segments and oxidative cross-linking of cysteine pairs to identify residues that are near each other in space . Propensity for cross-linking was determined for pairs of homologously placed cysteines in the two subunits of the dimer at all 54 possible positions . Also, combinations of cysteines were identified that readily oxidized to join heterologous segments within or between monomers . These patterns of cross-linking were used to develop a model for the three-dimensional structure of the transmembrane domain in which the four transmembrane segments are helices associated in a bundle, with stronger interactions near the periplasm and weaker interactions near the cytoplasm . The striking similarity of this model to a model for the transmembrane domain of chemoreceptor Tar, derived using the same experimental strategy, strengthens the notion that a combination of comprehensive cysteine substitutions and analysis of patterns of disulfide cross-linking is sufficient to deduce a detailed three-dimensional structure for a transmembrane domain. Nucleic Acids Res, 1994 Nov 25, 22(23), 4922 - 31 Construction and characterization of a bacterial artificial chromosome library of Sorghum bicolor; Woo SS et al.; The construction of representative large insert DNA libraries is critical for the analysis of complex genomes . The predominant vector system for such work is the yeast artificial chromosome (YAC) system . Despite the success of YACs, many problems have been described including: chimerism, tedious steps in library construction and low yields of YAC insert DNA . Recently a new E.coli based system has been developed, the bacterial artificial chromosome (BAC) system, which offers many potential advantages over YACs . We tested the BAC system in plants by constructing an ordered 13,440 clone sorghum BAC library . The library has a combined average insert size, from single and double size selections, of 157 kb . Sorghum inserts of up to 315 kb were isolated and shown to be stable when grown for over 100 generations in liquid media . No chimeric clones were detected as determined by fluorescence in situ hybridization of ten BAC clones to metaphase and interphase S.bicolor nuclei . The library was screened with six sorghum probes and three maize probes and all but one sorghum probe hybridized to at least one BAC clone in the library . To facilitate chromosome walking with the BAC system, methods were developed to isolate the proximal ends of restriction fragments inserted into the BAC vector and used to isolate both the left and right ends of six randomly selected BAC clones . These results demonstrate that the S . bicolor BAC library will be useful for several physical mapping and map-based cloning applications not only in sorghum but other related cereal genomes, such as maize . Furthermore, we conclude that the BAC system is suitable for most large genome applications, is more 'user friendly' than the YAC system, and will likely lead to rapid progress in cloning biologically significant genes from plants. Biochim Biophys Acta, 1994 Nov 22, 1219(3), 592 - 600 Enhancement of bacterial transcription initiation in vitro by the 74 kDa subunit of human general transcription factor IIF (RAP74); Chibazakura T et al.; The human general transcription factor IIF (TFIIF) is required for an accurate transcription initiation by RNA polymerase II and shares some analogous features with the sigma subunit of bacterial RNA polymerase . As an attempt to analyze the function of TFIIF, we examined its effect on bacterial transcription in vitro . TFIIF significantly enhanced the initiation of transcription by the bacterial RNA polymerase while other general transcription factors, TATA-binding protein, TFIIB, and TFIIE, did not . The enhancement of the bacterial transcription was ascribed to the 74 kDa subunit of TFIIF (RAP74) . RAP74 had an activity of enhancing the binding of the bacterial RNA polymerase to the promoter . The enhancing activity of RAP74 depended on a low molar ratio of the RNA polymerase to the template DNA . The action of RAP74 in the bacterial transcription may be related to a possible regulatory role of RAP74 in the eukaryotic transcription initiation. Virology, 1994 Nov 15, 205(1), 93 - 103 Bacterial expression of human respiratory syncytial viral phosphoprotein P and identification of Ser237 as the site of phosphorylation by cellular casein kinase II; Mazumder B et al.; The phosphoprotein P gene of human respiratory syncytial virus has been cloned and the protein expressed in Escherichia coli . The expressed protein was soluble, unphosphorylated, and constituted approximately 10% of the total bacterial protein . Electrophoretic and antigenic analyses demonstrated the identity of the recombinant protein with viral P protein and P protein synthesized in reticulocyte lysates . Purified recombinant P protein was efficiently phosphorylated in vitro by purified native as well as recombinant casein kinase II (CKII) or by the CKII activity in uninfected cell extracts . Through deletions and site-directed mutagenesis, the site of CKII phosphorylation was mapped to a single serine residue (Ser237) near the C-terminal end of the P protein. EMBO J, 1994 Nov 15, 13(22), 5330 - 7 Detecting the ability of viral, bacterial and eukaryotic replication proteins to track along DNA; Tinker RL et al.; The phage T4 gene 45 protein (gp45), Escherichia coli beta and the eukaryotic proliferating cell nuclear antigen (PCNA) function in replication as processivity factors of their corresponding DNA polymerases . The T4 gp45 also functions as the transcriptional activator that connects expression of viral late genes to DNA replication . DNA tracking is an essential component of the replication and transcription regulatory functions of T4 gp45 . The ability of gp45, beta and PCNA to track along DNA has been analyzed by photocrosslinking . Each of these proteins must be loaded onto DNA by a species-specific assembly factor . For gp45 and beta, the density of traffic along DNA is determined by a dynamic balance between continuous protein loading and unloading, and is also dependent on interaction with the conjugate single-stranded DNA binding protein. Biochemistry, 1994 Nov 15, 33(45), 13199 - 207 Exploring the microtubule-binding region of bovine microtubule-associated protein-2 (MAP-2): cDNA sequencing, bacterial expression, and site-directed mutagenesis; Coffey RL et al.; A 1.1 kilobase fragment of bovine microtubule-associated protein-2 (MAP-2) cDNA coding for bovine MAP-2 microtubule-binding region (MTBR) was sequenced . Relative to mouse, rat, and human MAP-2, we observed striking preservation of primary structure, even beyond the sequence and spacing of the three nonidentical peptide repeats responsible for microtubule-binding interactions . For further analysis of microtubule-MAP interactions using site-directed mutagenesis, we developed a bacterial expression system coding for the MT-binding fragment of MAP-2 starting at the thrombin cleavage site (position 1629) and continuing to the C-terminus . This MT-binding fragment was purified to homogeneity by taking advantage of the unusual heat-stability and isoelectric properties of this cytomatrix component . We found that the MT-binding domain readily promoted tubulin polymerization, and the critical tubulin concentration was reduced in the presence of this recombinant protein . Because a second repeated sequence analogue can promote tubulin polymerization as well as displace the MT-binding region of MAP-2, this study was designed to learn more about the importance of each repeated sequence in MT binding . Accordingly, we mutated the first and third sequences to resemble the second repeated sequence, thereby generating the mutants designed m12-m2-m3, m1-m32, and m12-m2-m32 . These recombinant proteins bound with an affinity comparable to or slightly better than equal concentrations of wild-type MT-binding fragment . Likewise, when the first or third sequence was replaced by an exact copy of the second octadecapeptide repeat, there was little, if any, increase in binding affinity, as reflected in the ability of mutant MT-binding fragments to promote tubulin polymerization.(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1994 Nov 11, 269(45), 28294 - 9 Effect of mutations in Escherichia coli glnL (ntrB), encoding nitrogen regulator II (NRII or NtrB), on the phosphatase activity involved in bacterial nitrogen regulation; Kamberov ES et al.; We examined the effects of mutations in glnL, encoding the signal-transducing kinase/phosphatase nitrogen regulator II (NRII), on the regulated phosphatase activity involved in nitrogen regulation . With wild-type NRII, this phosphatase activity was only observed in the presence of the signal transduction protein II (PII) . Three different glnL mutations result in altered NRII proteins that had phosphatase activity in the absence of PII . The most active of these contained an alteration of the site of NRII autophosphorylation, histidine 139, to asparagine (H139N) . The phosphatase activity of the NRII-H139N protein was further stimulated by the PII protein and by ATP . This suggests that the PII protein is not directly involved in a catalytic step of the regulated phosphatase activity but rather plays a regulatory role . We also measured the effect on the regulated phosphatase activity of alterations at conserved residues in the kinase/phosphatase domain of NRII and the effect of deleting the non-conserved N-terminal domain of NRII . For this we used fusion proteins containing the Escherichia coli maltose-binding protein (MBP) linked to the protein of interest . A protein consisting of MBP linked to wild-type NRII was a less active kinase than was wild-type NRII but in the presence of PII had wild-type phosphatase activity . A protein consisting of MBP linked to just the C-terminal domain of wild-type NRII had kinase activity but lacked phosphatase activity . Alterations at the highly conserved residues Asp-287, Gly-289, and Gly-291 in NRII affected both activities . A fusion of MBP to the NRII-H139N protein lacked kinase activity but had phosphatase activity in the absence of PII . Thus, while the kinase and phosphatase activities of NRII could be genetically separated, some of the highly conserved residues in the C-terminal domain of NRII (Asp-287, Gly-289, Gly-291) are apparently important for both activities. N Engl J Med, 1994 Nov 3, 331(18), 1181 - 7 A controlled trial of intravenous immune globulin for the prevention of serious bacterial infections in children receiving zidovudine for advanced human immunodeficiency virus infection . Pediatric AIDS Clinical Trials Group; Spector SA et al.; BACKGROUND . Serious bacterial infections are common in children infected with the human immunodeficiency virus (HIV) . Studies performed before zidovudine became standard therapy found that intravenous immune globulin decreases the number of serious bacterial infections in these children . We designed a multicenter study to evaluate the efficacy of intravenous immune globulin in children with advanced HIV infection who were receiving zidovudine . METHODS . In a double-blind trial 255 children between 3 months and 12 years of age who had the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex were randomly assigned to receive either intravenous immune globulin (400 mg per kilogram of body weight) (n = 129) or placebo (0.1 percent albumin) (n = 126) every 28 days . All children received 180 mg of zidovudine per square meter of body-surface area orally four times daily . Treatment assignment was stratified according to whether the patients had a history of one or more serious bacterial infections, had previously been treated with zidovudine, or were currently receiving prophylaxis with trimethoprim-sulfamethoxazole . The median length of follow-up was 30.6 months . RESULTS . The estimated two-year rates of serious bacterial infections with confirmed pathogens were 16.9 percent for the immune globulin group and 24.3 percent for the placebo group (relative risk, 0.60; 95 percent confidence interval, 0.35 to 1.04; P = 0.07) . The treatment effect was seen primarily among the 174 children who were not receiving trimethoprim-sulfamethoxazole prophylaxis at entry; the estimated two-year rates of infection were 11.3 percent for the immune globulin group and 26.8 percent for the placebo group (relative risk, 0.45; 95 percent confidence interval, 0.22 to 0.91; P = 0.03) . For the 81 children who were receiving trimethoprim-sulfamethoxazole prophylaxis initially, the rates were 27.7 percent in the immune globulin group and 17.7 percent in the placebo group (relative risk, 1.26; 95 percent confidence interval, 0.44 to 3.66; P = 0.67) . The two-year survival was similar in the two groups: 79.2 percent among immune globulin recipients and 75.4 percent among placebo recipients (P = 0.41) . CONCLUSIONS . In children with advanced HIV disease who are receiving zidovudine, intravenous immune globulin decreases the risk of serious bacterial infections . However, this benefit is apparent only in children who are not receiving trimethoprim-sulfamethoxazole as prophylaxis. Appl Environ Microbiol, 1994 Nov, 60(11), 3959 - 64 Plant and algal interference in bacterial beta-D-galactosidase and beta-D-glucuronidase assays; Davies CM et al.; Several commonly occurring freshwater and marine plants and algae were screened for beta-D-galactosidase and beta-D-glucuronidase activities by using a 60-min enzyme assay based on the hydrolysis by these enzymes of 4-methylumbelliferyl-beta-D-galactoside and 4-methylumbelliferyl- beta-glucuronide, respectively . All freshwater plant extracts tested showed beta-D-galactosidase activity several at relatively high levels, and a number also showed beta-D-glucuronidase activity . A number of the macroalgae showed no activity of either enzyme, but those showing beta-D-galactosidase activity also showed beta-D-glucuronidase activity . The majority of microalgae showed some beta-D-galactosidase activity, but few showed beta-D-glucuronidase activity . Further studies, using the commercial Colilert test and the marine water formulation of Colilert, revealed that 2 of 11 of the microalgal species and several of the plant extracts tested caused positive reactions . It was concluded that several plant extracts and algae could significantly interfere with the detection of coliform bacteria and Escherichia coli with the use of rapid assays, on the basis of their production of beta-D-galactosidase and beta-D-glucuronidase, respectively . The significance of the plant and algal interferences in tests such as Colilert is dependent on the levels of enzymes released under natural conditions, the dilution which they may undergo, and the numbers of algal cells present . This also applies to interferences in rapid enzyme assays.(ABSTRACT TRUNCATED AT 250 WORDS) Kyobu Geka, 1994 Nov, 47(12), 1026 - 8 {Left ventricular-right atrial shunt with bacterial and rheumatic endocarditis of tricuspid valve}; Nagashima K et al.; A 14-year-old boy who was operated on for left ventricular-right atrial shunt with bacterial and rheumatic endocarditis of the tricuspid valve (TV) is reported . The patient was pointed out perimembranous ventricular septal defect (VSD) at 4-month old and had complaint of occasional high fever for a half year . The echocardiography showed a vegetation at the TV . The vegetation did not disappear despite of administration of antibiotics for 2 weeks, and pulmonary embolism occurred . Then, the patient underwent resection of the vegetation and direct closure of VSD . The left ventricular-right atrial shunt was formed by the direct communication between VSD below TV annulus and the perforation of septal leaflet of TV adhered to the ventricular septum due to bacterial and rheumatic endocarditis . Earlier operation is needed for this type of case because antibiotics is not effective for the valvular disease. Arch Surg, 1994 Nov, 129(11), 1184 - 90 Quantitative and morphologic analysis of bacterial translocation in neonates; Go LL et al.; OBJECTIVE: To elucidate the mechanisms of bacterial translocation in animals fed a conventional formula by correlating transmucosal bacterial passage in vitro with the structural characteristics of the neonatal intestinal mucosa . DESIGN: Newborn rabbits were randomized to receive a conventional formula or breast milk . Bacterial translocation to the mesenteric lymph nodes, liver, and spleen was quantitated after 7 days, and transmucosal passage of bacteria was measured in vitro using the Ussing chamber . The mucosal membranes were examined by light, transmission electron, and confocal laser scanning microscopy . RESULTS: Bacterial passage was rarely seen in the breast milk-fed animals in contrast to the formula-fed animals . Unlike the normal-appearing membranes from breast milk-fed animals, the epithelial cells of formula-fed animals were vacuolated but healthy, with normal polarization and microvillus border by confocal laser scanning microscopy . Villi of formula-fed animals were less densely packed than those of the breast milk-fed animals . Bacterial adhesion, internalization, and transmucosal passage were seen only in membranes from formula-fed animals . Transmission electron microscopy demonstrated bacteria incorporating into the epithelial surface through an active phagocytic process, with rearrangement of the actin cytoskeleton . Once internalized, these bacteria were seen within the cytoplasmic vacuoles and subsequently in the submucosa . No bacteria passed between epithelial cells . CONCLUSION: Morphological changes in the intestinal mucosa of formula-fed newborn rabbits may increase permeability to bacteria. Eur J Immunol, 1994 Nov, 24(11), 2855 - 62 Monoclonal IgA class-switch variants against bacterial surface antigens: molecular forms and transport into murine respiratory secretions; Steinmetz I et al.; The present study describes a new model for passive immunization of the respiratory tract with IgA in comparison to other isotypes . Monoclonal IgA-isotype-switch variants were isolated from different IgG-producing hybridoma clones specific for surface epitopes of bacterial respiratory tract pathogens . Analysis of the molecular form of the IgA variants revealed the simultaneous production of monomeric, dimeric and higher polymeric IgA by a single-cell line with predominance of the polymeric forms . The specificities of the IgA variants were identical to the parent IgG antibodies as demonstrated by inhibition experiments . The IgA variant antibodies were separated into monomers and polymers by gel filtration . Intravenous injection of the different molecular forms of IgA and of IgG into mice were used to investigate the transport characteristics of IgA into murine upper and lower respiratory tract secretions by the physiological route in comparison to IgG . Polymeric IgA variant, monomeric IgA variant and IgG were detected in immunologically active form in both nasal secretion and bronchoalveolar fluid as evidenced by binding to their antigens in an enzyme-linked immunosorbent assay . The relative contribution of the specific exogenous monoclonal IgA and monoclonal IgG to total IgA and IgG, respectively, was determined in secretions . Comparison of the secretion to serum transport ratios clearly indicates selective transport of polymeric IgA variant into nasal secretions relative to IgG parent antibody . Molecular and functional characteristics of the IgA variants make them ideal for passive mucosal immunization experiments and identification of protective epitopes in mucosal immunity. EMBO J, 1994 Nov 1, 13(21), 5195 - 202 A novel device of bacterial signal transducers; Ishige K et al.; The osmoregulatory expression of ompC and ompF in Escherichia coli is mediated by a pair of bacterial signal transduction proteins, EnvZ (sensory kinase) and OmpR (response regulator) . We isolated previously multicopy suppressors which can complement a defect in the phosphotransfer signal transduction caused by an envZ deletion mutation . Among such suppressors, arcB and barA are of particular interest because these gene products are unique in the sense that they contain both an autophosphorylated histidine site (or transmitter module) and a phospho-accepting aspartate site (or receiver module) in their primary amino acid sequences . Here we report that ArcB and BarA possess in the C-terminal region a phosphorylated histidine site which has never been noticed, in addition to the authentic one identified previously . This newly identified histidine in ArcB and BarA was demonstrated to play a crucial role in the observed multicopy suppression . Furthermore, it was demonstrated in vivo and in vitro for ArcB that the C-terminal domain containing the histidine can function as an alternative phosphodonor (or transmitter) . This novel type of sensory kinase was therefore revealed to contain two independent phosphodonor sites, together with a phospho-accepting site . These findings suggest that this unique feature of ArcB and BarA, in terms of the signaling modules, make it possible for these sensory kinases to function as dual-signaling transducers. J Histochem Cytochem, 1994 Nov, 42(11), 1435 - 41 Detection of intestinal flora-derived bacterial antigen complexes in splenic macrophages of rats; Kool J et al.; We studied the presence of bacterial antigens in rat tissues . We produced a monoclonal antibody (MAb 2E9) directed against intestinal flora-derived peptidoglycan-polysaccharide complexes from human and rat feces . With several immunological techniques, the specificity of 2E9 for this bacterial product was demonstrated . Using 2E9 in an immunohistological assay, we were able to show the presence of bacterial products in macrophages in the red pulp of spleens of conventional Lewis rats . However, we found no correlation between the development of the intestinal flora and positive spleen staining with MAb 2E9 . The results were confirmed by immunohistology with a previously described MAb 2-4 directed to muramyl dipeptide . Other lymphoid organs did not stain positively with 2E9 and 2-4 . Neonatal and young rats showed no staining of the spleen, but positivity could be induced by injecting peptidoglycan-polysaccharide complexes systemically . We conclude that bacterial fragments are present in splenic macrophages of conventional rats. Infect Immun, 1994 Nov, 62(11), 4831 - 7 Activation of human effector cells by different bacterial toxins (leukocidin, alveolysin, and erythrogenic toxin A): generation of interleukin-8; Konig B et al.; We analyzed the transcription and release of interleukin-8 (IL-8) from human polymorphonuclear granulocytes (PMNs) and a lymphocyte-monocyte-basophil (LMB) cell population stimulated for different time periods (30 min to 16 h) with pore-forming bacterial toxins (Panton-Valentine leukocidin {Luk-PV} and alveolysin {Alv}) as well as with the erythrogenic toxin A (ETA) as a superantigen . At high toxin concentrations (500 ng/10(7) cells), Luk-PV and Alv led to a decreased IL-8 generation from LMBs within the first 30 min; with PMNs, a slight increase in IL-8 release was observed . Under these conditions, stimulation with ETA did not lead to an altered cellular IL-8 release . At lower concentrations (5 and 0.5 ng/10(7) cells), all three toxins led to a continuous increase (over 16 h) in IL-8 release and IL-8 mRNA expression of PMNs and LMBs . Preincubation of the cells with the protein tyrosine kinase inhibitors lavendustin A and tyrphostin 25 led to a reduction of the toxin-mediated effects on IL-8 release and IL-8 mRNA expression when Luk-PV and Alv were used as stimuli . In contrast, IL-8 synthesis in cells which were stimulated with ETA was not influenced by protein tyrosine kinase inhibition . From our data, one may suggest that multiple pathways for IL-8 production are operative in human leukocytes. Infect Immun, 1994 Nov, 62(11), 4768 - 74 Composition and diversity of intestinal coliform flora influence bacterial translocation in rats after hemorrhagic stress; Katouli M et al.; Coliform bacteria are the most frequently reported bacteria to translocate after hemorrhage . We investigated the correlation between composition and diversity of the cecal coliform flora and the degree of translocation in a rat model of hemorrhagic stress . Two groups of nine rats each were bled to 60 and 50 mm Hg mean arterial blood pressure, respectively . A sham-operated group without bleeding (n = 9) and a noninstrumented group (n = 6) served as controls . From each rat, 40 coliform isolates from the cecum and up to 16 from positive mesenteric lymph node (MLN) cultures were tested with an automated biochemical fingerprinting method . The phenotypic diversity of coliforms in each cecal sample was calculated as Simpson's diversity index (DI), and similarities between bacterial types in different samples were calculated as population similarity coefficients . Three rats in the sham-operated group and seven in each of the bled groups showed bacterial translocation . Of the different biochemical phenotypes (BPTs) found in the cecum of bled rats (mean, 6.5 BPTs), only a few were detected in MLNs (mean, 1.9 BPTs per MLN), with Escherichia coli being the dominant species . The translocating E . coli strains were mainly of two BPTs . Rats showing no translocation either did not carry these strains or had a high diversity of coliforms in the cecum . Furthermore, translocation of these coliform types was independent of their proportion in the cecum . In bled rats, the diversity of coliforms (mean DI, 0.53) was significantly higher than that in control groups (mean DI, 0.30; P = 0.004), suggesting that hemorrhage stimulates an increase in diversity of cecal coliforms . Rats with similar coliform flora and subjected to the same treatment showed similar patterns of translocation . Our results suggest that the composition of the coliform flora is an important factor in translocation and that certain coliform strains have the ability to translocate and survive in MLNs more easily than others. Indian Pediatr, 1994 Nov, 31(11), 1337 - 43 Cranial ultrasonic assessment of infants with acute bacterial meningitis; Soni JP et al.; A prospective study was performed on 44 patients (newborn to one year old) with acute bacterial meningitis . Sonograms were obtained within 72 hours of diagnosis, and repeated on 7th, 14th or at an early date, if required . The spectrum of sonographic features of meningitis included normal scan (35.4%), echogenic sulci (63.7%) and parenchyma (29.5%), ventriculomegaly (59%), ventriculitis (35%), pseudo-porencephalic cyst (4.4%), extra axial fluid collection (4.4%), encephalomalacia (2.2%) and cerebral abscess (2.2%) in patients. Int J Food Microbiol, 1994 Nov, 23(3-4), 277 - 94 A dynamic approach to predicting bacterial growth in food; Baranyi J et al.; A new member of the family of growth models described by Baranyi et al . (1993a) is introduced in which the physiological state of the cells is represented by a single variable . The duration of lag is determined by the value of that variable at inoculation and by the post-inoculation environment . When the subculturing procedure is standardized, as occurs in laboratory experiments leading to models, the physiological state of the inoculum is relatively constant and independent of subsequent growth conditions . It is shown that, with cells with the same pre-inoculation history, the product of the lag parameter and the maximum specific growth rate is a simple transformation of the initial physiological state . An important consequence is that it is sufficient to estimate this constant product and to determine how the environmental factors define the specific growth rate without modelling the environment dependence of the lag separately . Assuming that the specific growth rate follows the environmental changes instantaneously, the new model can also describe the bacterial growth in an environment where the factors, such as temperature, pH and aw, change with time. J Microsc, 1994 Nov, 176 ( Pt 2), 132 - 42 Unstained and in vivo fluorescently stained bacterial nucleoids and plasmolysis observed by a new specimen preparation method for high-power light microscopy of metabolically active cells; Kellenberger E et al.; Microscope slides were coated with a layer of gelatin, the thickness of the gelatin increasing linearly along the long axis . The bacterial suspension is applied to the dried gelatin and covered by a coverslip . The medium is absorbed by the gelatin and thus the cells applied against the coverslip . By this method, cultures of concentrations below 10(8) cells/ml provide statistically relevant numbers for observation without prior concentration steps . It is easier to apply than the existing methods for the observation of bacterial nucleoids by phase contrast imaging . Because the cells are maintained in growing conditions the method is useful for the vital fluorescence DAPI-staining of various bacterial species and for observations of plasmolysis and its reversal at different physiological conditions and extracellular osmolalities . The previously generally assumed view that the plasmolytic changes of the cell morphology are immediate upon the hyperosmotic shock and are rapidly repaired when the cell is able to metabolize actively was confirmed; this is in contrast to some recent claims. Zhonghua Yi Xue Za Zhi (Taipei), 1994 Nov, 54(5), 373 - 5 Pneumoperitoneum caused by spontaneous bacterial peritonitis: a case report; Lai FC et al.; Spontaneous bacterial peritonitis (SBP) rarely presents as pneumoperitoneum . Only four such cases have been reported in the English literature . This report concerns a 65 year-old male patient with SBP and pneumoperitoneum . He has had a history of peptic ulcer . However, upon examination, no associated diseases or compromised immunity was detected . He presented with acute abdominal pain and subphrenic free air . An emergency laparotomy was performed, under the impression of a perforated peptic ulcer . Yet, no intraabdominal pathology except 200 ml of purulent ascites was found . The ascites culture yielded E . coli, B . fragilis, and P . aeruginosa confirming the diagnosis of SBP . Antibiotics which are sensitive to gas-forming bacteria should be prescribed before the result of the ascites culture is known. Gut, 1994 Nov, 35(11), 1648 - 52 Bacterial translocation in cirrhotic rats . Its role in the development of spontaneous bacterial peritonitis; Llovet JM et al.; Bacterial translocation occurs in ascitic cirrhotic rats, but its association with ascites infection is unknown . The aim of this study was to assess the relation between bacterial translocation and ascites infection in cirrhotic rats . Male Sprague-Dawley rats were induced to cirrhosis with intragastric CCl4 . Ascitic fluid, portal and peripheral blood, mesenteric lymph nodes, liver and spleen samples were cultured before death in those cirrhotic rats with less (group A) or more (group B) than 250 polymorphonuclear neutrophils/mm3 in ascitic fluid, as well as in healthy control rats . Histological examination of jejunum, ileum, and caecum was also performed . Bacterial translocation occurred in 45% of ascitic rats (without differences between groups A and B), but in 0% controls (p = 0.01) . Bacterial translocation was associated with positive ascitic fluid culture in 60% of the cases . In all of them the same bacterial species was isolated in both mesenteric lymph node and ascitic fluid . Submucosal caecal oedema (100%), ileal lymphangiectasia (41%), and caecal inflammatory infiltrate (41%) occurred in ascitic rats, the last being associated with ascitic fluid positive culture (p = 0.04) . These results suggests that bacterial translocation occurs frequently in ascitic cirrhotic rats, and may play a permissive, but not unique, part in a number of ascites infections . Whether histological changes seen in cirrhotic ascitic rats favour bacterial translocation remains to be elucidated. AORN J, 1994 Nov, 60(5), 796, 799 - 805 Bacterial carriage on the fingernails of OR nurses; Wynd CA et al.; This study provides statistically significant data that demonstrate that chipped fingernail polish or fingernail polish worn longer than four days fosters increased numbers of bacteria on the fingernails of OR nurses after surgical hand scrubs . There were no significant correlations between fingernail length and the numbers of bacterial colonies on the fingernails of the study groups tested after performing a standard surgical hand scrub . A convenience sample of 102 perioperative nurses with either freshly polished fingernails; chipped fingernail polish; or natural, polish-free fingernails participated . The data suggest OR nurses can wear fresh fingernail polish on healthy fingernails without risking increased bacterial counts. Mol Biol Evol, 1994 Nov, 11(6), 911 - 20 Unusual pattern of bacterial ice nucleation gene evolution; Edwards AR et al.; Bacterial ice nucleation activity (INA+ phenotype) can be traced to the product of a single gene, ina . A remarkably sparse distribution of this phenotype within three bacterial genera indicates that the ina gene may have followed an unusual evolutionary path . Southern blot analyses, coupled with assays for ice-nucleating ability, revealed that within four bacterial species an ina gene is present in some strains but absent from others . Results of hybridization experiments using DNA fragments that flank the ina gene suggested that the genotypic dimorphism of ina may be anomalous . A phylogenetic analysis of 16S ribosomal RNA gene sequences from a total of 14 ina+ and ina- bacterial strains indicated that the ina+ bacteria are not monophyletic but instead phylogenetically interspersed among ina- bacteria . The relationships of ina+ bacteria inferred from ina sequence did not coincide with those inferred from the 16S data . These results suggest the possibility of horizontal transfer in the evolution of bacterial ina genes. Res Microbiol, 1994 Nov-Dec, 145(9), 647 - 50 Bacterial protein kinases that recognize tertiary rather than primary structure? Saier MH Jr. While all characterized eukaryotic protein kinases that phosphorylate hydroxy aminoacyl residues in proteins recognize primary structure, certain bacterial protein kinases are proving to recognize tertiary structure . It is proposed that these latter enzymes evolved independently of the superfamily of the former protein kinases and that their modes of target protein recognition and action are entirely different. Insect Mol Biol, 1994 Nov, 3(4), 247 - 52 High-level expression of the bacterial opd gene in Drosophila melanogaster: improved inducible insecticide resistance; Benedict MQ et al.; The bacterial parathion hydrolase gene (opd) was expressed in transformed D . melanogaster under the control of an hsp70 promoter . Transformed lines carrying chimaeric genes designed for either cytoplasmic or secretory expression exhibited high- or low-level heat-shock-inducible transient resistance to paraoxon respectively . Greatest levels of resistance occurred approximately 12-16 h after heat shock and well after periods of maximal transcription . Insecticide resistance conferred by the cytoplasmic form of opd is expressed as a semidominant trait. Intensive Care Med, 1994 Nov, 20 Suppl 4, S12 - 6 Diagnosis of bacterial infection in the ICU: general principles; Langer M et al.; Diagnosis and treatment of infection is a common procedure in the clinical management of patients in the ICU . Infection in the ICU is an important area for study, but requires well-defined and proven diagnostic criteria . The diagnosis of infection, like any diagnosis, is based on probability, and diagnostic criteria are therefore selected according to the physician's objectives and the acceptable margin of error . It is easier to diagnose correctly a full-blown, severe bacterial infection than one that is just beginning, and the same criteria cannot be used to identify accurately both conditions . We should diagnose an infectious complication at the time it needs treatment, but there is often a lack of clear objectives in the diagnostic process, and up to now, few reliable criteria have been available . Before considering the sensitivity and specificity of single diagnostic procedures it is important to trace the evolution of the infection . The problem may be approached in two steps, by describing or defining (i) the minimum level of severity of a probable infection which requires/justifies specific treatment as the first end-point of the diagnosis, and (ii) the ways the diagnosis may be confirmed using the best available procedure (which might not be always available or applicable in all cases in the short term). J Endod, 1994 Nov, 20(11), 535 - 7 Influence of apical enlargement on bacterial infection during treatment of apical periodontitis; Yared GM et al.; This study sought to evaluate the influence of apical enlargement and 1-wk calcium hydroxide dressing on bacterial infection of root canals . Sixty single-rooted teeth were used . Half of these teeth were prepared to a size 25 file and the other half to a size 40 file . Then the root canals were dressed with calcium hydroxide for 1 wk . Bacterial sampling showed significant reduction of bacterial growth during the treatment . No statistically significant difference was noted between the size 25 and 40 file groups after instrumentation, and after 1-wk calcium hydroxide dressing. Rev Inst Med Trop Sao Paulo, 1994 Nov-Dec, 36(6), 531 - 7 Dot-enzyme-linked immunosorbent assay (Dot-ELISA) for detection of pneumococcal polysaccharide antigens in pleural fluid effusion samples . Comparison with bacterial culture, counterimmunoelectrophoresis and latex agglutination; Requejo HI et al.; A dot-enzyme-linked immunosorbent assay (Dot-ELISA) for pneumococcal antigen detection was standardized in view of the need for a rapid and accurate immunodiagnosis of acute pneumococcal pneumonia . A total of 442 pleural fluid effusion samples (PFES) from children with clinical and laboratory diagnoses of acute bacterial pneumonia, plus 38 control PFES from tuberculosis patients and 20 negative control serum samples from healthy children were evaluated by Dot-ELISA . The samples were previously treated with 0.1M EDTA pH 7.5 at 90 degrees C for 10 min and dotted on nitrocellulose membrane . Pneumococcal omniserum diluted at 1:200 was employed in this assay for antigen detection . When compared with standard bacterial culture, counterimmunoelectrophoresis and latex agglutination techniques, the Dot-ELISA results showed relative indices of 0.940 to sensitivity, 0.830 to specificity and 0.760 to agreement . Pneumococcal omniserum proved to be an optimal polyvalent antiserum for the detection of pneumococcal antigen by Dot-ELISA . Dot-ELISA proved to be a practical alternative technique for the diagnosis of pneumococcal pneumonia. N Engl J Med, 1994 Oct 27, 331(17), 1122 - 8 Impaired function of macrophage Fc gamma receptors and bacterial infection in alcoholic cirrhosis; Gomez F et al.; BACKGROUND . Bacterial infection is a frequent and often fatal complication in patients with cirrhosis . Macrophages play an important part in the host defense against infection because their Fc gamma receptors recognize antibody-coated bacteria . METHODS . We prospectively studied macrophage Fc gamma-receptor function in vivo and in vitro in 49 patients with alcoholic cirrhosis, 10 alcoholics without cirrhosis, and 20 normal volunteers . RESULTS . The clearance of IgG-sensitized autologous red cells was decreased in 37 of the 49 patients with cirrhosis but in none of the subjects without cirrhosis . In the 49 patients clearance was inhibited by a mean (+/- SE) of 47 +/- 3 percent at 1 hour and 53 +/- 3 percent at 1 1/2 hours, as compared with the clearance in the normal controls (P < 0.001) . The impairment of macrophage Fc gamma-receptor-dependent clearance correlated with the degree of liver insufficiency but not with age, sex, nutritional status, HLA haplotype, or the presence of circulating immune complexes . The clearance of unsensitized and heat-altered autologous erythrocytes was normal . In vitro recognition of IgG-sensitized red cells by monocytes from the patients was not significantly decreased . During a two-year follow-up period, 11 patients had severe bacterial infections, and in 4 they were fatal . The mean clearance of IgG-sensitized red cells in these 11 patients (half-time, 126.2 +/- 22 hours) was significantly impaired, as compared with that in the 38 patients without severe infection (half-time, 32.2 +/- 18 hours, P < 0.001) . CONCLUSIONS . The function of macrophage Fc gamma receptors is impaired in patients with alcoholic cirrhosis, and this impairment probably contributes to the high incidence of bacterial infections among such patients. Biochim Biophys Acta, 1994 Oct 18, 1219(2), 277 - 84 Macromolecular crowding effects on the interaction of DNA with Escherichia coli DNA-binding proteins: a model for bacterial nucleoid stabilization; Murphy LD et al.; DNA-binding protein fractions from exponential and stationary phase cell extracts of E . coli were isolated by affinity chromatography on native DNA-cellulose . The ability of these fractions to convert DNA into a readily-sedimented form was compared in the absence or presence of added polymers . In the absence of polymers, large amounts of the proteins were required . In the presence of polyethylene glycol or polyvinylpyrrolidone, much smaller amounts of the DNA-binding proteins were required, indicating a macromolecular crowding effect from these polymers . The enhanced binding under crowded conditions appears to resolve a paradox between the cellular abundance of the DNA-binding proteins and the amounts required in earlier in vitro studies . The 'histone-like' protein HU from the DNA-binding protein fraction was preferentially incorporated into the pelleted DNA in the presence of polymers . Purified HU at roughly similar amounts caused a similar conversion of DNA to a readily-sedimentable ('condensed') form . Crowding-enhancement of DNA condensation by promoting the binding of proteins to the DNA provides a model for the stabilization of systems such as the bacterial nucleoid or kinetoplast DNA. Biochemistry, 1994 Oct 18, 33(41), 12405 - 15 Binding sites of quinones in photosynthetic bacterial reaction centers investigated by light-induced FTIR difference spectroscopy: binding of chainless symmetrical quinones to the QA site of Rhodobacter sphaeroides; Breton J et al.; Light-induced FTIR QA-/QA difference spectra corresponding to the photoreduction of the primary quinone acceptor QA have been obtained for Rhodobacter sphaeroides RCs reconstituted with chainless symmetrical quinones in order to study the influence of the side chain and of molecular asymmetry on the binding of natural quinones to the QA site . The main vibrational modes of the quinones in vivo were obtained by analysis of the isotope effects induced by 18O substitution on the carbonyls and by comparison with the IR absorption spectra of the isolated quinones . For isolated 2,3-dimethoxy-5,6-dimethyl-1,4-benzoquinone (MQ0), 2,3,5,6-tetramethyl-1,4-benzoquinone (duroquinone, DQ), and 2,3-dimethyl-1,4-naphthoquinone (DMNQ), the IR spectra together with mass spectroscopy data of partially 18O labeled quinones show that the labeling of one carbonyl leads to only a minor shift of the vibrational frequency of the opposite carbonyl . This observation demonstrates an essentially uncoupled behavior of the two C = O groups . Upon reconstitution of QA-depleted RCs with these symmetrical quinones, the double-difference spectra calculated from the QA-/QA spectra of the 18O-labeled and unlabeled quinones reveal a splitting of the quinone C = O modes . This splitting and the frequency downshift of the C = O vibrations upon binding to the QA site are comparable to those previously reported for the C = O modes of quinones containing an isoprenoid (Q8, Q6, Q1) or a phytyl chain (vitamin K1) {Breton, J., Burie, J.-R., Berthomieu, C., Berger, G., & Nabedryk, E . (1994) Biochemistry 33, 4953-4965} . This observation demonstrates that the replacement of the side chain by a methyl group does not impair the asymmetrical bonding interactions of the two quinone carbonyls with the protein . This asymmetry is traceable to the two distinct amino acid residues which have been proposed, on the basis of X-ray structural studies, to form hydrogen bonds with the carbonyls of the quinone . The close analogy between the double-difference spectra calculated for RCs reconstituted either with vitamin K1 or with DMNQ shows that the phytyl chain of vitamin K1 imparts no specific constraint on the geometry of the menaquinone head group in its binding site for both the neutral and the semiquinone state . In contrast, the double-difference spectra calculated for RCs reconstituted either with MQ0 or with Q6 (or Q1) exhibited significant differences in the relative amplitudes of the bands assigned to the mixed C = O and C = C modes of the neutral quinones.(ABSTRACT TRUNCATED AT 400 WORDS) Eur J Biochem, 1994 Oct 15, 225(2), 765 - 71 Structure elucidation and biosynthesis of 31-methylhopanoids from Acetobacter europaeus . Studies on a new series of bacterial triterpenoids; Simonin P et al.; Apart from a mixture of bacteriohopanetetrols already found in other Acetobacter species, four new 3 beta-methylhopanoids have been isolated from Acetobacter europaeus . All of them present an ether linkage between a bacteriohopanetetrol or a bacteriohopanepentol and a carbapseudopentose moiety often found in bacterial hopanoids . Three of these ethers were shown by comparison with synthetic reference hopanoids to posess a supplementary methyl group at C31 . This novel series of methylhopanoids may be the precursor of yet unidentified molecular fossils found in sediments . {methyl-2H3}Methionine was efficiently incorporated into the 31-methylhopanoids with retention of all three deuterium atoms in the transferred methyl group . This labelling pattern might be consistent with a rather rarely found methylation reaction of an enol. J Biol Chem, 1994 Oct 14, 269(41), 25315 - 20 Modeling the bacterial protein toxin, pneumolysin, in its monomeric and oligomeric form; Morgan PJ et al.; Pneumolysin is a member of the family of related bacterial thiol-activated toxins, which share structural similarities and a proposed common cytolytic mechanism . Currently the molecular mechanism of membrane damage caused by these toxins remains a matter of controversy . A prerequisite for defining this mechanism is a detailed knowledge of the monomeric and oligomeric pneumolysin structures . We present for the first time details of the monomeric structure of a thiol-activated toxin, pneumolysin . Electron microscope images of metal-shadowed pneumolysin monomers show an asymmetric molecule composed of four domains . We have studied the conformation of pneumolysin monomer by low resolution hydrodynamic bead modeling procedures . The bead model dimensions and shape are derived solely from the electron micrographs . The bead model has been evaluated in terms of the predicted solution properties, which in turn have been compared to the experimental values of the sedimentation coefficient, s(20,w)0, obtained by analytical ultracentrifugation and the intrinsic viscosity, {eta} . Pneumolysin oligomers, observed as ring- and arc-shaped structures, were also examined by electron microscopy . Metal shadowing and negative staining methods were used to establish the overall dimensions of the oligomer and were used to produce a morphological model for the oligomer, incorporating monomer subunits based on the hydrodynamic bead model. Nature, 1994 Oct 13, 371(6498), 578 - 86 The crystal structure of the bacterial chaperonin GroEL at 2.8 A; Braig K et al.; The crystal structure of Escherichia coli GroEL shows a porous cylinder of 14 subunits made of two nearly 7-fold rotationally symmetrical rings stacked back-to-back with dyad symmetry . The subunits consist of three domains: a large equatorial domain that forms the foundation of the assembly at its waist and holds the rings together; a large loosely structured apical domain that forms the ends of the cylinder; and a small slender intermediate domain that connects the two, creating side windows . The three-dimensional structure places most of the mutationally defined functional sites on the channel walls and its outward invaginations, and at the ends of the cylinder. J Biol Chem, 1994 Oct 7, 269(40), 24870 - 7 Forskolin specifically inhibits the bacterial galactose-H+ transport protein, GalP; Martin GE et al.; Forskolin is a potent inhibitor of mammalian passive glucose transporters . Here we show that forskolin is a remarkably specific inhibitor of energized D-galactose transport by the GalP sugar-H+ symport protein of Escherichia coli . Surprisingly, it does not inhibit transport of L-arabinose or D-xylose by the related E . coli AraE and XylE transporters, even though the amino acid sequences of their proteins are 30-64% identical to GalP and to the mammalian GLUT family . However, unlike GLUT1, photoactivation of the {3H}forskolin-GalP complex fails to incorporate radioactivity covalently into the protein, in contrast to the effective incorporation of radioactivity from {3H}cytochalasin B into both proteins . However, 3-{125I}iodo-4-azidophenethylamido-7-O-succinyldesacetylforskol in ({125I}APS-forskolin), which labels GLUT1, is a potent labeling reagent for GalP and, to a lesser extent, for AraE . The appropriate sugar substrates of each transporter protect it against the {125I}APS-forskolin . Equilibrium binding studies using membranes from an E . coli strain that overexpresses GalP reveal a single set of high affinity binding sites for {3H}forskolin with a Kd of 1.3-1.4 microM, probably forming a 1:1 complex, compared with a value of 7.5 microM for GLUT1 . Sugar substrates of GalP and cytochalasin B displace forskolin from the protein . The nonhomologous sugar-H+ symporters for L-rhamnose (RhaT), L-fucose (FucP) and lactose (LacY) in E . coli are insensitive to forskolin . Forskolin and {125I}APS-forskolin, therefore, constitute novel probes for exploring the structure-activity relationship of the bacterial GalP protein . GalP will provide an excellent model for the human glucose transporters and for elucidating the molecular basis of subtle differences in substrate and inhibitor recognition by individual members of this widespread family of transport proteins. Am J Dent, 1994 Oct, 7(5), 293 - 5 Evaluation and treatment of bacterial microleakage; Cox CF; Recent data have shown a correlation between pulp response and material biocompatibility when microleakage is prevented . Considering vital dentin, the dentist must keep in mind that it is an extension of the pulp, presenting the first line of response to the consequences of microleakage . Healing of the dental pulp is ensured when a clinical seal is provided to prevent bacterial microleakage . Microleakage is a biological phenomenon with several important clinical implications . The affective component clinically presents as dentin postoperative hypersensitivity due to the episodic hydrodynamic fluid movement within the tubule complex . The effective component is the disease process which signals the dentin and pulp complex to respond by deposition of sclerotic and reparative dentin deposition . Uncontrolled microleakage generally produces patient pain as well as to later allow bacterial infection resulting in recurrent caries and pulp inflammation . The dynamic, permeable nature of the dentin interface allows residual bacteria to proliferate following cavity preparation . Following sterile operative procedures, bacteria have been shown to percolate into the cavosurface margin resulting in recurrent caries and pulp inflammation . This report considers biological data emphasizing the clinical necessity to provide a hermetic seal to prepared dentin to prevent both dentin hypersensitivity and pulp inflammation. Am J Physiol, 1994 Oct, 267(4 Pt 1), G687 - 95 Mechanisms of transit of lipid mediators of inflammation and bacterial peptides across intestinal epithelia; Riehl TE et al.; The transit of two lipid mediators of inflammation, leukotriene B4 (LTB4) and prostaglandin E2 (PGE2), and a formylated peptide produced by intestinal bacteria, N-formylmethionylleucylphenylalanine (FMLP), across Caco-2 cell monolayers was characterized and compared with the transit of mannitol, a hexose known to cross epithelial monolayers by paracellular pathways . The permeability of less mature low-resistance ( < 200 ohm.cm2) monolayers to all four test compounds was similar, but as monolayers matured and the transmonolayer resistance increased, the transit of LTB4, PGE2, FMLP, and mannitol decreased to different degrees, resulting in a selectivity of permeability to the four test compounds in the order LTB4 > PGE2 > mannitol > FMLP . The transit of all four test compounds across Caco-2 cell monolayers was bidirectional, nonsaturable, and energy independent . A small portion of the added LTB4 was incorporated into the cells, whereas the other three compounds were not . Thus the transit of PGE2, mannitol, and FMLP across Caco-2 monolayers appears to be solely by the paracellular pathway, whereas the transit of LTB4 also involves the paracellular pathway but may also involve diffusion through the cell membrane and around tight junctions. Am J Kidney Dis, 1994 Oct, 24(4), 569 - 74 Serum from continuous ambulatory peritoneal dialysis patients with acute bacterial peritonitis inhibits in vitro erythroid colony formation; Stevens JM et al.; During episodes of acute infection there is a reduced response to epoetin therapy . It is well known that "endogenous pyrogens," such as interleukin-1 (IL-1) and tumor necrosis factor, inhibit erythropoiesis when administered exogenously . To determine whether there is a relationship between these observations, serum samples were obtained from nine patients with chronic renal failure maintained by continuous ambulatory peritoneal dialysis, during and after recovery from bacterial peritonitis, to study the effect of circulating factors on erythropoiesis . Normal human bone marrow-derived erythroid progenitors were cultured in vitro in 5% and 10% patient serum . Depression of the growth of late progenitors, colony-forming units-erythroid (at 10% serum, P = 0.005; 95% confidence intervals, 6.2 and 24.4, respectively), was observed but there was no effect on the earlier progenitors, burst-forming units-erythroid (at 10% serum, P = 0.7; 95% confidence intervals, -18.5 and 13, respectively) . The effect was not prevented by antisera to IL-1 . Similarly, when added to cultures, IL-1 inhibited the colony-forming units-erythroid and the effect was abrogated by IL-1 antisera . These findings suggest that a circulating soluble factor that is inhibitory to erythropoiesis and may contribute to loss of response to epoetin therapy, is present in cases of peritonitis in continuous ambulatory peritoneal dialysis patients. J Exp Med, 1994 Oct 1, 180(4), 1225 - 33 Bordetella pertussis filamentous hemagglutinin interacts with a leukocyte signal transduction complex and stimulates bacterial adherence to monocyte CR3 (CD11b/CD18); Ishibashi Y et al.; Bordetella pertussis, the causative agent of whooping cough, adheres to human monocytes/macrophages by means of a bacterial surface-associated protein, filamentous hemagglutinin (FHA) and the leukocyte integrin, complement receptor 3 (CR3, alpha M beta 2, CD11b/CD18) . We show that an FHA Arg-Gly-Asp site induces enhanced B . pertussis binding to monocytes, and that this enhancement is blocked by antibodies directed against CR3 . Enhancement requires a monocyte signal transduction complex, composed of leukocyte response integrin (alpha? beta 3) and integrin-associated protein (CD47) . This complex is known to upregulate CR3 binding activity . Thus, a bacterial pathogen enhances its own attachment to host cells by coopting a host cell signaling pathway. J Infect Dis, 1994 Oct, 170(4), 854 - 61 Fc gamma receptor IIa (CD32) heterogeneity in patients with recurrent bacterial respiratory tract infections; Sanders LA et al.; Fc gamma RIIa (CD32) is the sole IgG Fc receptor capable of interaction with human IgG2, the main IgG subclass of bacterial capsular polysaccharides . The two genetically determined allotypes of human Fc gamma RIIa, Fc gamma RIIa-R131 and IIa-H131 alleles, have functionally different reactivities with human IgG2 . The capacity of polymorphonuclear leukocytes (PMNL) homozygous for Fc gamma RIIa-H/H131 for IgG2 opsonized bacteria is significantly higher than phagocytosis by PMNL homozygous for Fc gamma RIIa-R/R131, independent of the Fc gamma RIIb-NA1/NA2 (CD16) allelic polymorphism . To test the clinical significance of these Fc gamma R polymorphisms, Fc gamma RIIa and Fc gamma RIIIb phenotypes of 48 children with recurrent bacterial respiratory tract infections were determined . Fc gamma RIIa-H/H131 was less than half that observed in 123 healthy adults (P = .01) . IgG2 responses were low in 25 of 48 patients after immunization with pneumococcal vaccine . These results suggest that Fc gamma RIIa polymorphism may contribute to increased susceptibility to infections with encapsulated bacteria in a childhood population with low IgG2 anti-carbohydrate antibodies. Infect Immun, 1994 Oct, 62(10), 4404 - 10 Epithelial glucosphingolipid expression as a determinant of bacterial adherence and cytokine production; Svensson M et al.; D-Threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) is a structural analog of ceramide that inhibits glucosylation of this molecule and thus of glucosphingolipid (GSL) expression by living cells . In this study, we used PDMP to slow the synthesis of the globoseries of GSLs (globo-GSLs) (derived from the precursor Gal alpha 1-4Gal beta 1-4Glc-ceramide) by cultured human kidney and large intestinal epithelial cells . The aim was to deplete the cells of receptors for P-fimbriated Escherichia coli and to examine the effects on the bacterially induced cytokine response . The mammalian cells (A498, HT-29, and Caco2) were cultured in the presence of PDMP in order to deplete them of GSLs . The cells were then subjected to GSL analysis or used to test bacterial adherence and cytokine production . The globo-GSLs were identified by thin-layer chromatography . Bacterial adherence was quantitated by microscopy, and interleukin-6 secretion was quantitated by the B9 bioassay . The interaction of bacteria with the globo-GSLs was studied by using E . coli strains and recombinant clones expressing P fimbriae . E . coli strains expressing type 1 fimbriae binding to mannose-containing glycoproteins were used as controls . PDMP treatment was found to reduce the content of the globo-GSLs in mammalian cells and the adherence of P-fimbriated E . coli to these cells . In contrast, PDMP treatment had no effect on the adherence of type 1-fimbriated E . coli or their activation of cytokine production by A498 cells . P-fimbriated E . coli elicited an interleukin-6 response in the A498 cells; this response was reduced after treatment with PDMP . The results emphasize the role of GSLs as receptors for P-fimbriated E . coli and for the cytokine response elicited by attaching bacteria. Environ Res, 1994 Oct, 67(1), 98 - 107 Superoxide anion production in response to bacterial lipopolysaccharide and fungal spores implicated in organic dust toxic syndrome; Shahan TA et al.; High amounts of fungal spores, bacteria, and bacterial endotoxin have been found in dust associated with the poorly characterized syndrome, organic dust toxic syndrome (ODTS) . As part of an ongoing investigation to determine the etiopathogenesis for ODTS, this study has focused on activation of guinea pig bronchial alveolar lavage (BAL) cells as evidenced by the production of superoxide anion in response to fungal spores and lipopolysaccharide (LPS) . Fungal spores from Aspergillus candidus, Aspergillus terreus, Aspergillus niger, Aspergillus fumigatus, Eurotima amstelodami, Penicillium spinulosum, and Cladosporium cladosporioides were all shown to increase superoxide anion production, each with different potencies . LPS stimulated little superoxide anion production in BAL cells, but when cells were pretreated with LPS prior to stimulation with fungal spores, superoxide anion production was increased over that induced by either spores or LPS alone . These results suggest that the inhalation of LPS together with fungal spores could possibly provoke abnormal lung pathologies. New Microbiol, 1994 Oct, 17(4), 345 - 62 Bacterial protein toxins acting on the cell cytoskeleton; Donelli G et al.; A number of bacterial protein toxins are known to exert their cytotoxic activity via a modification of cytoskeletal components . Some toxins induce the ADP-ribosylation of actin whereas others interact with the cytoskeleton by an unknown mechanism . Understanding the mode of action of such toxins at cellular level could provide useful information on their role in vivo as virulence factors. J Am Dent Assoc, 1994 Oct, 125(10), 1369 - 72 Coronal leakage: bacterial penetration through obturated canals following post preparation; Gish SP et al.; Coronal leakage of bacteria from saliva into root canal filling materials is a potential cause of failure . This problem may be more pronounced when only a small volume of obturating material remains in the canal, such as after post preparation . In this study, coronal leakage of bacteria through unsealed, apically obturated canals was investigated using a new in vitro model system. Inflammation, 1994 Oct, 18(5), 499 - 510 Administration of large doses of vitamin C does not decrease oxidant-induced lung lipid peroxidation caused by bacterial-independent acute peritonitis; Demling R et al.; Acute zymosan-induced peritonitis in rats produces lung inflammation and lipid peroxidation . The effect of this process on plasma and lung tissue ascorbic acid was determined, as was the effect of infusing 150 mg/kg of ascorbic acid immediately after zymosan on the degree of lung insult . Ascorbic acid levels were significantly decreased in plasma and lung tissue at 24 h after zymosan, and lung tissue conjugated diene and neutrophil content was also significantly increased . Vitamin C infusion increased postzymosan plasma levels by 50% over normal control levels . However, lung tissue ascorbic acid was still decreased, and no decrease in the lung injury process was noted . Added ascorbic acid also did not prevent a decrease in plasma vitamin E with the peritonitis . We conclude that the amount of ascorbic acid given in this study did not diminish the lung oxidant inflammatory changes . An insufficient dose or inadequate time for plasma ascorbic acid to equilibrate with the lung cytosol are possible explanations for the lack of attenuation of lung oxidant stress. Acta Anaesthesiol Scand, 1994 Oct, 38(7), 699 - 703 Possible errors in diagnosis of bacterial sinusitis in tracheal intubated patients; Westergren V et al.; Bacterial sinusitis is recognized as a complication of prolonged nasotracheal intubation . Verification of the diagnosis, however, requires culture findings from a "true" sample from the maxillary antrum . A "true" antral sample can only be obtained after passing either the nasal or oral mucosae with their rich aerobic and anaerobic bacterial flora . No properly controlled study has been made into the problem of contaminated samples in bacterial sinusitis complicating prolonged nasotracheal intubation . The design of the study was a case series in an intensive care unit . The patients had either had endotracheal tube or tracheal cannula for seven days or were septic, having an endothracheal tube or a tracheal cannula and therefore due to an investigation of infection foci . Antral cultures obtained via the canine fossa were compared with oral cultures to exclude the contribution of oral bacterial contamination and thereby to improve the reliability of diagnosis based on culture results . The contamination rate was lowered significantly from 67% to 5% when an area of free bone was prepared in the canine fossa thus avoiding instrument contact with the oral mucosa . A high contamination risk can be dealt with by adjusting the technique of sampling thereby increasing the culture reliability. Biotechniques, 1994 Oct, 17(4), 714 - 6, 718 A vector, pHisGal, allowing bacterial production of proteins fused to a hexahistidine-tagged Gal4 DNA-binding domain; Schmitz ML et al.; A vector, pHisGal, was constructted that allows isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible expression of the DNA-binding domain of the yeast transcription factor Gal4 in Escherichia coli . Protein sequences to be tested for transcription regulatory potential in conjunction with the Gal4 DNA-binding domain can be inserted in-frame into a multiple cloning site at the C terminus of Gal4 . A hexahistidine sequence fused to the N terminus of Gal4 allows efficient purification of the bacterially produced protein by affinity chromatography on nickel nitrilotriacetic acid (Ni-NTA) columns . Purified Gal4 fusion proteins produced in E . coli showed DNA-binding activity in electrophoretic nobility shift assays and were highly active in cell-free transcription assays from higher eukaryotic cells. Eur Respir J, 1994 Oct, 7(10), 1759 - 64 Bacterial adhesion to oropharyngeal and bronchial epithelial cells in smokers with chronic bronchitis and in healthy nonsmokers; Riise GC et al.; Bacterial adhesion is probably a prerequisite for colonization of mucous membranes, but adhesion to the bronchial mucosa has not been studied in detail . We investigated adhesion of respiratory pathogens to bronchial epithelial cells, and asked whether chronic bronchitis had an influence on bacterial adhesion . Oropharyngeal and bronchial cells were collected during bronchoscopy from 14 healthy nonsmokers, 22 smokers with nonobstructive chronic bronchitis, and 19 smokers with chronic bronchitis and chronic obstructive pulmonary disease (COPD) . Patients with a forced expiratory volume in one second (FEV1) less than 50% predicted were excluded . Adhesion of highly adherent test strains of H . influenzae and S . pneumoniae to these cells were studied . The test strains of H . influenzae and S . pneumoniae were found to adhere well to both oropharyngeal and bronchial cells . H . influenzae showed a higher degree of adhesion both to ciliated and goblet cells from the patients with nonobstructive bronchitis than to cells from the healthy nonsmokers . No corresponding difference was found for S . pneumoniae . The patients with COPD did not differ from the controls in their adhesion values . Our results indicate that bacterial adhesion is of importance for the colonization and retention of H . influenzae in the human airways . For S . pneumoniae the role of adhesion is more uncertain. Acta Paediatr Jpn, 1994 Oct, 36(5), 485 - 8 The fluctuations of neuron-specific enolase (NSE) levels of cerebrospinal fluid during bacterial meningitis: the relationship between the fluctuations of NSE levels and neurological complications or outcome; Inoue S et al.; Neuron-specific enolase (NSE) is one of the glycolytic enzymes distributed exclusively in neurons . It was measured serially in the cerebrospinal fluid (CSF) of 10 children with bacterial meningitis during the illness using radio-immunoassay . The relationship between CSF-NSE levels and neurological complications or outcome was examined . CSF-NSE levels were significantly higher in the patients with bacterial meningitis than in the patients with the other central nervous system (CNS) infectious diseases, suggesting that CNS damage in those patients with bacterial meningitis was exacerbated . As CSF-NSE levels increased to above 25 ng/mL in the acute phase, all patients except one had subdural effusion . In those patients whose CSF-NSE level rose again during the illness, CNS complications or sequelae occurred . CSF-NSE may be a useful prognostic factor for predicting CNS damage in childhood bacterial meningitis. Neuroendocrinology, 1994 Oct, 60(4), 418 - 25 Systemically administered histamine H1 and H2 receptor antagonists do not block the ACTH response to bacterial lipopolysaccharide and interleukin-1; Perlstein RS et al.; The administration of lipopolysaccharide (LPS) results in the activation of the hypothalamic-pituitary-adrenal axis (HPAA) . We recently reported that the participation and interaction of LPS-induced proinflammatory cytokines were obligatory for the stimulation of adrenocorticotropic hormone (ACTH) release by LPS . LPS and LPS-derived cytokines also stimulate the release of histamine (HA) . HA is a known hypothalamic neurotransmitter and activates the HPAA . Therefore, to elucidate the role of HA in LPS- and cytokine-induced ACTH release, we evaluated the effects of several HA H1 and H2 receptor antagonists on the ACTH response to LPS, recombinant human interleukin-1 alpha (rhIL-1 alpha) and HA in mice . Although all 3 of the H1 receptor antagonists administered (mepyramine (MEP), diphenhydramine (DPH) or promethazine (PMZ) were able to block the 10-min ACTH response to HA, only PMZ (a less selective H1 receptor antagonist than MEP) was able to reduce the LPS- or rhIL-1 alpha-induced ACTH responses . Ranitidine, a powerful and selective H2 receptor antagonist, had little effect on the LPS- and rhIL-1 alpha-induced ACTH responses, while metiamide (MET), a much less potent first-generation H2 receptor antagonist, substantially diminished ACTH release . The greater effectiveness of PMZ, in contrast to MEP or DPH, probably relates to the ability of phenothiazine derivatives to inhibit non-HA-dependent pathways involved in the stimulation of the HPAA by cytokines; the same may be true of MET.(ABSTRACT TRUNCATED AT 250 WORDS) Clin Transplant, 1994 Oct, 8(5), 479 - 84 Soluble interleukin-2 receptor monitoring during bacterial and viral infections in liver transplant recipients: a comparative evaluation; Rossi SJ et al.; We evaluated the significance of serial sIL-2R serum levels as a differential marker of immune activation during bacterial versus viral infections in liver transplant recipients . A comparative evaluation of sIL-2R levels was performed in 76 liver transplant recipients (51 pediatric and 27 adult) during bacterial versus viral infections at 7 days prior to infection diagnosis (DAY-7), the day of diagnosis (DAY 0), peak sIL-2R level and at the end of therapy (END) . There were no significant elevations at any time point during bacterial infections in either adult or pediatric transplant recipients . However, adult recipients demonstrated significant elevations during viral infections when comparing DAY-7 to PEAK (3840 +/- 830 vs 7225 +/- 2814 p = 0.03), with PEAK levels significantly higher during viral versus bacterial infections in this population (7225 +/- 2814 vs 4195 +/- 1819) . Pediatric recipients demonstrated similar increases in sIL-2R serum levels during viral infections from DAY-7 to PEAK (4932 +/- 887 vs 11323 +/- 2794 p = 0.0012) . Significant decreases from PEAK to END were noted during viral infections in both adult and pediatric recipients (7225 +/- 2814 vs 2911 +/- 1376 p = 0.01 and 11323 +/- 2794 vs 5214 +/- 2403 p = 0.006) . Pediatric recipients had higher mean sIL-2R levels than adult recipients at all time points during viral infections . In conclusion, significant elevations in mean sIL-2R serum levels were observed during viral but not bacterial infections in pediatric and adult liver transplant recipients . This suggests that serial sIL-2R monitoring is a valuable immunologic marker of viral pathogenesis and may be useful in monitoring the progression of viral infections as well as response to antiviral therapy. Shock, 1994 Oct, 2(4), 262 - 6 Does intravenous glutamine prevent bacterial translocation in hemorrhagic shock? Premaratne S, Masuda E, Nishida S, Suehiro A, McNamara JJ. Bacterial translocation across the gut wall may be associated with insult to the latter . In this situation, intestinal flora can enter the blood stream and lymph nodes and be transported to other organs . Glutamine is a nonessential amino acid not presently included in total parenteral nutrition (TPN) preparations . The use of glutamine-enriched TPN in the rat has resulted in a significant reduction in bacterial translocation . This study attempted to evaluate the role of glutamine in preventing bacterial translocation following hemorrhagic shock in a rat model . Forty Sprague-Dawley rats were equally divided into two groups . The controls were given TPN solution, while the treated group had glutamine instead of the standard alanine present in TPN . Hemorrhagic shock was induced in both groups and blood cultures were performed . Glutamine-treated rats did not show a significant difference in survival suggesting that it is of no particular value in severe hemorrhagic shock in rats. Microb Pathog, 1994 Oct, 17(4), 203 - 12 Apoptosis induced by bacterial pathogens; Chen Y et al.; Programmed cell death is part of normal development and homeostasis . Apoptosis induced by bacteria appears to contribute to infectious diseases . Some bacteria produce toxins to kill host cells by the same pathway, apoptosis, through different mechanisms including pore formation, protein synthesis inhibition or adenylate cyclase activity . Other bacterial pathogens' mechanisms to induce apoptosis, for example, that of S . flexneri, remain to be elucidated . How the bacterial toxins or the bacteria interact with eukaryotic cell-death-related genes and then possibly trigger a cell-death program would make an interesting study . The understanding of the mechanism of apoptosis induced by bacteria could be important in the development of therapy and prevention of infectious diseases. Neurol Med Chir (Tokyo), 1994 Oct, 34(10), 697 - 9 Cerebral bacterial aneurysm and indications for cerebral angiography in infective endocarditis; Yamada M et al.; Six infective endocarditis patients who developed cerebral bacterial aneurysm were reviewed to clarify the indications and timing for cerebral angiography to achieve early detection of unruptured aneurysms . All cerebral bacterial aneurysms were confirmed either angiographically or at autopsy . All patients were treated conservatively . Four patients died due to ruptured aneurysm . Four of the six patients showed the signs and symptoms of cerebral and/or systemic embolism, followed by rupture or detection of cerebral bacterial aneurysm . Prodromal signs and symptoms of embolism in patients with infective endocarditis should be considered as indicators for cerebral angiography to detect cerebral bacterial aneurysms before rupture. Mutat Res, 1994 Oct 1, 310(1), 103 - 12 Are rogue cells an indicator of cancer risk due to the action of bacterial restriction endonucleases? Ahuja YR, Obe G. Cytogenetic surveys in normal individuals have occasionally shown the occurrence of cells with multiple chromosome-type aberrations in some of the subjects . These cells, which are rare, have been termed as rogue cells . Rogue cells, which have been observed worldwide, have a mysterious nature . It has been suggested that they may give rise to cancer . Various mechanisms have been considered for the causation of the rouge-cell phenomenon in the past but none of them appears to be fully justified . In this paper we propose their occurrence due to the action of bacterial restriction endonucleases. Gene, 1994 Sep 30, 147(2), 169 - 77 Characterization of the mouse Rep-3 gene: sequence similarities to bacterial and yeast mismatch-repair proteins; Liu K et al.; The mouse Rep-3 gene is transcribed divergently from the same promoter region as the dihydrofolate reductase-encoding gene and has a deduced amino-acid sequence that shares identity with the bacterial protein, MutS, which is involved in DNA mismatch repair . We have cloned Rep-3, mapped it and sequenced all of the known exons and their intron junction sequences . We find that the open reading frame is considerably larger than initially reported and that the most abundant form of Rep-3 mRNA encodes a protein of 123 kDa . The gene spans at least 134 kb and consists of 26 exons, including several alternatively spliced exons . All of the exon/intron junctions match the expected consensus sequences with the exception of the splice junctions for intron 6, which has AT and AC dinucleotides instead of the usual GT and AG bordering the exon sequences . The junction sequences for this intron share consensus sequences with three intron sequences from other genes, thereby helping to establish an alternative consensus sequence. Biochemistry, 1994 Sep 27, 33(38), 11554 - 62 5-Chloro{1,4-13C}levulinic acid modification of mammalian and bacterial porphobilinogen synthase suggests an active site containing two Zn(II); Jaffe EK et al.; 5-Chloro{1,4-13C}levulinic acid ({1,4-13C}CLA) is an active site-directed inactivator of porphobilinogen synthase (PBGS) . PBGS asymmetrically condenses two molecules of 5-aminolevulinic acid (ALA) which are called A-side ALA and P-side ALA in reference to their fates as the acetyl and propionyl halves of the product . {1,4-13C}CLA modifies bovine PBGS at the A-side ALA binding site . The C4 chemical shift indicates an intact keto moiety; the C1 chemical shift indicates a deprotonated carboxyl group . In contrast, {1,4-13C}CLA modification of Escherichia coli PBGS is heterogeneous and occurs preferentially at the P-side ALA binding site . The C1 chemical shifts indicate substantially deprotonated carboxylic acid groups . For one of four observed forms of {1,4-13C}CLA-modified E . coli PBGS, an analog of the P-site Schiff base is found . Bovine and E . coli PBGS contain two different zincs, ZnA and ZnB . Past results placed ZnA near A-side ALA . {1,4-13C}CLA modifies E . coli PBGS at Cys119 or Cys129, which is part of a four-cysteine cluster implicated in binding ZnB . This result places ZnB near P-side ALA . E . coli PBGS binds a third type of divalent metal, MgC or MnC, which is found to have no significant effect on the 13C NMR spectrum of the {1,4-13C}CLA-modified protein. Biochim Biophys Acta, 1994 Sep 21, 1208(1), 151 - 6 Bacterial expression and site-directed mutagenesis of two critical residues (tyrosine-151 and lysine-155) of human placental NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase; Ensor CM et al.; NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) catalyzes the first step in the catabolic pathway of the prostaglandins . This enzyme oxidizes the 15-hydroxyl group of prostaglandins to produce 15-keto metabolites which are usually biologically inactive . In this study the cDNA for human placental 15-PGDH was expressed in Escherichia coli and the recombinant enzyme was purified to homogeneity and characterized . The N-terminus of the recombinant protein was sequenced and found to be identical with the known amino-acid sequence of 15-PGDH . Determinations of Km and Vmax values for a number of the prostaglandins and NAD+ indicate that the recombinant enzyme does not appear to be kinetically different from the human placental enzyme . Site-directed mutagenesis was used to examine the importance of two residues which are highly conserved in the short-chain dehydrogenases which are known to be related to 15-PGDH . Tyrosine-151 was changed to phenylalanine and serine while lysine-155 was changed to glutamine and leucine . Western blot analysis indicated that the mutant and wild-type proteins were expressed at the similar levels . However, all of the mutant proteins were found to be inactive . These results indicate that both tyrosine-151 and lysine-155 are required for 15-PGDH activity. J Immunol, 1994 Sep 15, 153(6), 2642 - 52 CD14-dependent activation of protein kinase C and mitogen-activated protein kinases (p42 and p44) in human monocytes treated with bacterial lipopolysaccharide; Liu MK et al.; To investigate mechanisms of mononuclear phagocyte cell signaling, the effects of bacterial LPS on protein kinase activities in normal human peripheral blood monocytes were examined . Incubation of intact monocytes with LPS brought about time- and concentration-dependent increases in myelin basic protein (MBP) phosphotransferase activity in high speed supernatants of cell lysates . Anion-exchange chromatography on Mono Q demonstrated that LPS treatment resulted in two principal peaks of stimulated MBP kinase activity . Evidence was obtained to indicate that the first eluted peak of MBP kinase activity is accounted for by p42 and p44 mitogen-activated protein (MAP) kinases . Thus, 1) MBP kinase activity within peak 1 was quantitatively precipitated by anti-MAP kinase Abs, 2) the enzyme effectively phosphorylated a specific peptide substrate, 3) peak 1 contained proteins of subunit size M(r) 42,000 and M(r) 44,000 that reacted specifically with anti-MAP kinase Abs, and that 4) were recognized by anti-phosphotyrosine Abs only after stimulation of cells with LPS . Studies of the second peak of LPS-stimulated MBP kinase activity indicate that it is an isoform of protein kinase C (PKC) because: 1) enzyme activity was quantitatively immunoprecipitated by anti-PKC Abs, 2) the activity of the enzyme was potently and selectively inhibited by a specific peptide modeled on the autoinhibitory domain of PKC, and 3) the presence of a protein of subunit size M(r) 80,000 recognized by anti-PKC Abs . Because the second peak of MBP kinase activity (like the first) was active in the absence of added calcium and in the presence of 2 mM EGTA, it appears to be a type II, calcium-independent isoform of PKC . Abs to CD14 completely abrogated LPS-induced activation of both Mono Q peaks of MBP phosphotransferase activity . These results indicate that LPS coordinately activates both an apparently calcium-independent PKC and MAP kinase in mononuclear phagocytes and these responses appear to be initiated by signaling through the cell surface receptor, CD14. J Immunol Methods, 1994 Sep 14, 174(1-2), 195 - 202 Quantification of bacterial phagocytosis by flow cytometry and spectrofluorimetry; Plasman N et al.; Bacterial phagocytosis is a cardinal function of phagocytes . We describe a simple procedure to easily quantify this function using fluoresceinated bacteria . Non-ingested bacteria and those adsorbed to the cell membrane are eliminated by an enzymatic procedure . Only macrophages with ingested fluorescent bacteria are detected, thereby permitting an accurate quantification of the phagocytic process by both spectrofluorimetric measurement and flow cytometric analysis. Nurs Times, 1994 Sep 14-20, 90(37), 43 - 4 Patient education . Diagnosis and treatment of bacterial vaginosis; Temple CA; This paper continues our series looking at some of the information patients may require with regard to common health problems . Providing information that is comprehensible is an important part of the nurse's role . It is vital that information, usually provided for particular specialties, is made available to all health-care workers and patients. Biochim Biophys Acta, 1994 Sep 13, 1219(1), 179 - 83 Cloning and nucleotide sequence of a bacterial cytochrome P-450VD25 gene encoding vitamin D-3 25-hydroxylase; Kawauchi H et al.; The gene encoding an enzyme that catalyzes the hydroxylation at position 25 of vitamin D-3 was cloned from an actinomycete strain, Amycolata autotrophica, by use of a host-vector system of Streptomyces lividans . The amino acid sequence deduced from the nucleotide sequence revealed that this enzyme, tentatively named P-450VD25, contains several regions of strong similarity with amino acid sequences of cytochromes P-450 from a variety of organisms, primarily in the regions of an oxygen-binding site and a heme ligand pocket . Especially, P-450VD25 shows end-to-end similarity in amino acid sequence to P-450dNIR of Fusarium oxysporum and P-450SU2 of Streptomyces griseolus . The recombinant S . lividans strain containing the P-450VD25 gene on a multicopy plasmid converted vitamin D-3 in the medium into 25-hydroxyvitamin D-3 at a maximum yield of 10%. FEBS Lett, 1994 Sep 12, 351(3), 360 - 4 Generation of the topa quinone cofactor in bacterial monoamine oxidase by cupric ion-dependent autooxidation of a specific tyrosyl residue; Matsuzaki R et al.; The quinone of 2,4,5-trihydroxyphenylalanine (topa), recently identified as the covalently bound redox cofactor in copper amine oxidases, is encoded by a specific tyrosine codon . To elucidate the mechanism of its formation, the recombinant phenylethylamine oxidase of Arthrobacter globiformis has been overproduced in Escherichia coli and purified in a Cu(2+)-deficient form . The inactive precursor enzyme thus obtained was dramatically activated upon incubation with Cu2+, concomitantly with the formation of the topa quinone at the position corresponding to Tyr382, occurring in the tetrapeptide sequence highly conserved in this class of enzymes . The topa quinone was produced only under aerobic conditions, but its formation required no external enzymatic systems . These findings demonstrate the Cu(2+)-dependent autooxidation of a specific tyrosyl residue to generate the topa quinone cofactor. Neuroimmunomodulation, 1994 Sep-Oct, 1(5), 300 - 7 ACTH response to a low dose but not a high dose of bacterial endotoxin in rats is completely mediated by corticotropin-releasing hormone; Schotanus K et al.; In experimental animals and humans, bacterial endotoxin activates the hypothalamus-pituitary-adrenal (HPA) axis . The pathways by which endotoxin stimulates adrenocorticotropic hormone (ACTH) and corticosterone secretion are uncertain . In the present study we compared the role of hypothalamic corticotropin-releasing hormone (CRH) in the activation of the HPA axis by a low (2.5 micrograms/kg) and a high (2.5 mg/kg) dose of bacterial endotoxin . Two experimental models were applied using chronically cannulated male Wistar rats . In the first model, rats were subjected to lesions of the hypothalamus that interrupted dorsal, lateral and frontal input to the median eminence (anterolateral deafferentation) or to sham operation and rats were used 7 days later . Before and at hourly intervals after endotoxin (2.5 micrograms/kg i.p.), blood samples were taken for the determination of plasma ACTH and corticosterone concentrations . Deafferentation of the hypothalamus strongly attenuated the elevations in plasma ACTH and corticosterone concentrations by a low dose of endotoxin compared to the responses in sham-operated animals . The second model involved passive immunization to CRH using a monoclonal antibody to rat/human CRH (PFU83) . PFU83 (90 nmol/rat) abolished the elevation of plasma ACTH concentrations and attenuated corticosterone responses to a low dose of endotoxin (2.5 micrograms/kg i.p.) compared to that in control IgG-treated rats . Since the corticosterone responses to endotoxin were less effectively inhibited by the antibody than the ACTH responses, we postulate that non-ACTH-dependent mechanisms may contribute to the corticosterone response to endotoxin.(ABSTRACT TRUNCATED AT 250 WORDS) J Surg Res, 1994 Sep, 57(3), 408 - 15 Bacterial translocation after intraperitoneal implantation of rubber fragments in the splenectomized rat; Guo W et al.; The aim of the present study was to determine the influence of splenectomy on the incidence of enteric bacterial translocation in rats with intraperitoneal rubber drain implantation . Male Sprague-Dawley rats (250-300 g) underwent splenectomy or sham operation 7 days prior to the intraperitoneal implantation of rubber drain fragments (7 cm2) . Bacterial translocation was measured 2 days after rubber drain implantation . The incidence of bacterial translocation was significantly higher in the group with intraperitoneal rubber drain implantation plus sham splenectomy than in the groups with sham splenectomy plus sham implantation, splenectomy plus sham implantation or splenectomy plus rubber drain implantation . An increase in ileal permeability of 125I-human serum albumin was induced by intraperitoneal rubber drain implantation and ameliorated by splenectomy . Splenectomy also improved the impaired intestinal motility induced by intraperitoneal rubber drain implantation . Histological examination revealed a preserved normal mucosal architecture in splenectomized rats . Thus, splenectomy reduced the rate of enteric bacterial translocation induced by intraperitoneal biomaterial implantation. J Pediatr, 1994 Sep, 125(3), 401 - 2 Bacterial tracheitis as a complication of tonsillectomy and adenoidectomy; Eid NS et al.; We describe two patients in whom bacterial tracheitis developed shortly after elective tonsillectomy and adenoidectomy . Bacterial tracheitis has not previously been reported in this clinical setting . Prompt recognition is essential if a fatal outcome is to be avoided . The cause remains uncertain, but the outcome is good if timely treatment is instituted. J Immunol, 1994 Sep 1, 153(5), 2072 - 81 Tat protein from HIV-1 binds to Mycobacterium avium via a bacterial integrin . Effects on extracellular and intracellular growth; Denis M; We examined the interaction between HIV-1 Tat protein and the opportunistic pathogen Mycobacterium avium . AIDS-associated strains of M . avium were shown to bind Tat protein quite avidly in an attachment assay . The attachment of M . avium to Tat was shown to occur via the integrin alpha 5 beta 1 present on the mycobacterial cell surface . M . avium strains were shown to bind the viral Tat protein with high affinity in a specific fashion (600 binding sites with a Kd of 1 to 5 nM) . M . avium coated with Tat protein were shown to be more infective for human alveolar macrophages than untreated M . avium . Other HIV-1 Ags had no such effects (e.g., p24, p17) . Examination of the cytokine profile of infected macrophages showed that M . avium-Tat complexes induced higher levels of TGF beta-1 (TGF beta 1) than M . avium alone or M . avium that had been in contact with other viral proteins . Conditioned media from HIV-1-infected H9 cells released a factor that enhanced M . avium intramacrophage growth, and was partially neutralized by an anti-Tat Ab . Finally, Tat protein (purified or present in conditioned media from infected cells) moderately enhanced the growth of M . avium strains in extracellular media, and exposure of M . avium to Tat protein in the presence of IL-6 enhanced the growth of AIDS-associated strains . These data argue for an interaction between the Tat viral product and the opportunistic pathogen M . avium which may contribute to the exquisite susceptibility of AIDS subjects to this pathogen. Immunopharmacology, 1994 Sep-Oct, 28(2), 163 - 70 Administration of the bacterial extract Broncho-Vaxom enhances radiation recovery and myelopoietic regeneration; Fedorocko P et al.; In the present study, we show that the bacterial extract Broncho-Vaxom (BV, 500 micrograms/mouse; free of endotoxin) has radiation recovery activity when administered i.p . 24 h before sublethal irradiation . In the postirradiation period (5-12 days), pretreatment of mice with BV induced significantly increased bone marrow cellularity and accelerated myelopoietic regeneration (committed progenitor granulocyte-macrophage colony-forming cells; GM-CFC) in the bone marrow compared with saline-treated controls . The earlier hemopoietic recovery in BV-injected mice was not associated with an increase in the number of bone marrow GM-CFC and CFU-S (colony-forming units-spleen) within 24 h after injection . Simultaneously, a significant diminution in bone marrow cellularity occurred . In addition, the percentage of both GM-CFC and CFU-S in the S-phase of the cell cycle was significantly increased 24 h after a single treatment . In our experiments colony stimulating activity (CSA) in the serum of treated mice was not observed within 24 h after injection . Administration of BV 24 h prior to lethal irradiation, resulted in an increase in the number of surviving mice . Combined administration of BV (24 h) and indomethacin (24 h and 3 h) to mice, prior to irradiation, caused an additional radioprotective effect . These results demonstrate that BV stimulates myelopoietic regeneration and suggest a mechanism by which this treatment protects mice from otherwise lethal irradiation. Rev Med Brux, 1994 Sep-Oct, 15(5), 317 - 21 {Bacterial endotoxins: a pathogenic factor in asthma?}; Michel O; Though the house dust mite allergen is considered as one of the most important factors in perennial asthma, several observations suggest that it could be but one of several factors leading to asthma . Exposure to bacterial endotoxins has been associated with some occupational chronic obstructive bronchial diseases . In the present work, endotoxins were measured in the house dust, and observations of the skin and bronchial challenge tests as well epidemiological studies suggest that they play a role in asthma. Neth J Med, 1994 Sep, 45(3), 93 - 100 Does long-term inhibition of gastric acid secretion with omeprazole lead to small intestinal bacterial overgrowth? Nelis GF, Engelage AH, Samson G. OBJECTIVE: Gastric acid secretion and small intestinal motility are the main mechanisms of defense against bacterial overgrowth of the proximal digestive tract . Bacterial colonization of the stomach during gastric acid inhibition has been documented, but is probably without clinical consequence . However, small intestinal bacterial overgrowth can have serious clinical implications with malabsorption and diarrhoea . METHODS: We prospectively investigated small intestinal bacterial overgrowth in 40 patients receiving long-term omeprazole treatment using the {14C}glycocholic breath test . Tests were performed before omeprazole treatment, after 6 weeks treatment with 40 mg o.m . and after 26 weeks treatment with 20 mg; in the test each patient served as his own control . RESULTS: Breath tests, using individual curves, peak values, time at which the peak appeared and the area under the curve, did not differ significantly during treatment from those before treatment . CONCLUSIONS: We conclude that long-term strong inhibition of gastric acid secretion does not lead to small intestinal bacterial overgrowth. Mol Cell Biochem, 1994 Sep, 138(1-2), 177 - 81 Common structure of the catalytic sites of mammalian and bacterial toxin ADP-ribosyltransferases; Okazaki IJ et al.; The amino acid sequences of several bacterial toxin ADP-ribosyltransferases, rabbit skeletal muscle transferases, and RT6.2, a rat T-cell NAD glycohydrolase, contain three separate regions of similarity, which can be aligned . Region I contains a critical histidine or arginine residue, region II, a group of closely spaced aromatic amino acids, and region III, an active-site glutamate which is at times seen as part of an acidic amino acid-rich sequence . In some of the bacterial ADP-ribosyltransferases, the nicotinamide moiety of NAD has been photo-crosslinked to this glutamate, consistent with its position in the active site . The similarities within these three regions, despite an absence of overall sequence similarity among the several transferases, are consistent with a common structure involved in NAD binding and ADP-ribose transfer. Mol Microbiol, 1994 Sep, 13(6), 939 - 53 The role of the pilus in recipient cell recognition during bacterial conjugation mediated by F-like plasmids; Anthony KG et al.; The effects of defined mutations in the lipopolysaccharide (LPS) and the outer membrane protein OmpA of the recipient cell on mating-pair formation in liquid media by the transfer systems of the F-like plasmids pOX38 (F), ColB2 and R100-1 were investigated . Transfer of all three plasmids was affected differently by mutations in the rfa (LPS) locus of the recipient cell, the F plasmid being most sensitive to mutations that affected rfaP gene expression which is responsible for the addition of pyrophosphorylethanolamine (PPEA) to heptose I of the inner core of the LPS . ColB2 transfer was more strongly affected by mutations in the heptose II-heptose III region of the LPS (rfaF) whereas R100-1 was not strongly affected by any of the rfa mutations tested . ompA but not rfa mutations further decreased the mating efficiency of an F plasmid carrying a mutation in the mating-pair stabilization protein TraN . An F derivative with a chloramphenicol acetyltransferase (CAT) cassette interrupting the traA pilin gene was constructed and pilin genes from F-like plasmids (F, ColB2, R100-1) were used to complement this mutation . Unexpectedly, the results suggested that the differences in the pilin sequences were not responsible for recognizing specific groups in the LPS, OmpA or the TraT surface exclusion protein . Other corroborating evidence is presented suggesting the presence of an adhesin at the F pilus tip. Zhonghua Yi Xue Za Zhi, 1994 Sep, 74(9), 552 - 5, 583-4 {Delayed fluid resuscitation induced bacterial translocation after lethal thermal injury: role of oxygen free radical injury of intestinal mucosa}; Yang HM et al.; The Present investigation was undertaken to examine the effect of delayed fluid resuscitation (DFR), after lethal thermal injury on oxygen free radical (OFR) injury of intestinal mucosa and its relationship to bacterial translocation . Four groups of gnotobiotic rats with 5 strains of bacteria were studied: sham injury group (control) (n = 6): early fluid resuscitation (EFR) group (n = 24) receiving resuscitation (Parkland) immediately after scald (40% TBSA, third degree); DFR group (n = 24) receiving resuscitation 6 hours after scald; treatment group (n = 12), rats with DFR receiving VitE and VitC treatment before resuscitation 12, 24, 48 and 72 hours after injury, animals (n = 6, at each point) were sacrificed . Plasma endotoxin level, mucosal SOD, GSHPx, MDA and diamine oxidase (DAO) of ileum were determined, and cultures of the mesenteric lymph nodes (MLN), liver, spleen, heart, lung, kidney and blood were done . The level of mucosal MDA and plasma endotoxin and the incidence of bacteria translocation (IBT) to tissues were significantly higher and mucosal SOD, GSHPx, DAO activity significantly lower in DFR group as compared with that in EFR group at most of the time points . In DFR group, mucosal MDA content was negatively correlated with mucosal DAO activity, which correlated positively with plasma endotoxin level and IBT . After treatment with VitE and VitC, mucosal MDA content was decreased, plasma endotoxin level and IBT were significantly decreased, and mucosal DAO activity was significantly increased . Our data indicated that DFR in cases of burn shock can result in OFR injury of intestinal mucosa, disrupting mucosal barrier and promoting translocation of intestinal bacteria and endotoxin. J Appl Physiol, 1994 Sep, 77(3), 1542 - 7 Prior exercise suppresses the plasma tumor necrosis factor response to bacterial lipopolysaccharide; Bagby GJ et al.; This study was initiated to determine the effect of physical exercise on the in vivo tumor necrosis factor-alpha (TNF) response to Escherichia coli lipopolysaccharide (LPS) . Rats familiarized with treadmill running and surgically implanted with vascular catheters were either not exercised or exercised to near exhaustion (mean run time of 102 +/- 13 min) before intravenous LPS challenge (1 mg/kg; lethality of dose is 10-20% in 24 h) . Compared with time-matched nonexercised control rats, exercised rats had increased heart rates, plasma lactate, and plasma corticosterone and decreased plasma glucose at the conclusion of exercise . In response to LPS, both groups became hypotensive, exhibited transient hyperglycemia, and sustained hyperlactacidemia . By 30 min post-LPS, plasma corticosterone levels were similar in the two groups . Nonexercised rats exhibited a normal plasma TNF response to LPS with the peak value (10,400 +/- 2,000 U/ml) occurring 90 min after LPS challenge . In contrast, the TNF response in rats exercised before LPS administration was blunted to 17% of the nonexercised group, with the peak occurring at an earlier time after LPS . Addition of recombinant murine TNF to postexercise plasma was fully expressed . The TNF response remained attenuated when LPS was administered up to 6 h after completion of exercise, but it returned to normal in rats allowed to recover for 24 h . The results demonstrate that exercise, perhaps as a stress modality, markedly suppresses the systemic TNF response that is normally observed in response to LPS challenge. Scand J Gastroenterol, 1994 Sep, 29(9), 833 - 40 Water-soluble ethylhydroxyethyl cellulose: a new agent against bacterial translocation from the gut after major liver resection; Wang X et al.; BACKGROUND: Bacterial translocation from the gut to extraintestinal sites has been demonstrated as a mechanism explaining bacterial infectious complications after various insults . METHODS: To explore the potential therapeutic value of water-soluble ethylhydroxyethyl cellulose (EHEC) . Its effects on macrophage phagocytic capacity, bacterial adherence on the intestinal surface, and bacterial growth were evaluated both in vivo and in vitro . RESULTS: Preoperative administration of EHEC reduced the incidence of bacterial translocation from the gut to mesenteric lymph nodes and blood and prevented overgrowth by enteric bacteria after 70% or 90% hepatectomy . Uptake of macrophages harvested from blood decreased after intravenous administration of EHEC . EHEC diminished the otherwise increased bacterial adherence on the intestinal surface induced by major liver resection . EHEC in bacterial cultures for over 1 h was capable of inhibiting bacterial growth and delaying bacterial DNA synthesis in vitro . CONCLUSIONS: The present study indicates that EHEC could be a potential agent for the prevention of gut-origin sepsis. Mol Microbiol, 1994 Sep, 13(5), 755 - 64 The bacterial phosphotransferase system: new frontiers 30 years later; Saier MH Jr et al.; In 1964, Kundig, Ghosh and Roseman reported the discovery of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) . Thirty years later, we find that the PTS functions not only as a sugar-phosphorylating system, but also as a complex protein kinase system that regulates a wide variety of metabolic processes and controls the expression of numerous genes . As a result of recent operon- and genome-sequencing projects, novel PTS protein-encoding genes have been discovered, most of which have yet to be functionally defined . Some of them appear to be involved in cellular processes distinct from those recognized previously . Fundamental aspects of past and current PTS research are briefly reviewed, and recent advances are integrated into conceptual pictures that provide guides for future research. Z Ernahrungswiss, 1994 Sep, 33(3), 239 - 43 {Comparative studies of the bacterial and mycological status of ecologically and conventionally grown crops}; Marx H et al.; Wheat and rye derived from conventional and from alternative or ecological production was examined for bacterial and fungal contamination . The overall bacterial and fungal contamination was lower than 10(7.7) germs/g . The amount and type of germs in the grain corresponded to typical contamination of fresh crop . No difference in germ contamination of both conventionally and alternatively grown wheat and rye could be found . No correlation between bacterial or fungal contamination and thousand-kernel-weight was detected. J Eukaryot Microbiol, 1994 Sep-Oct, 41(5), 503 - 10 Monoclonal antibody to a bacterial endonuclear symbiont Holospora cross reacts with proteins of contractile vacuole radial canals of Paramecium species; Dohra H et al.; A monoclonal antibody (mAb) IR-2-1 was raised against a 67-kDa protein purified from the macronucleus-specific bacterial symbiont Holospora obtusa of Paramecium caudatum . The mAb was found to react with two bands (31 and 67-kDa) on gels of H . obtusa . Indirect immunofluorescence microscopy showed that these antigens were distributed inside the cells . However, unexpectedly, this mAb also cross reacted with the radial arms of the contractile vacuole in P . caudatum, P . tetraurelia, P . multimicronucleatum, P . jenningsi and P . bursaria as well as with their cytoplasm . Immunoelectron microscopy showed that the antigens were located on the decorated spongiome of the radial arms . In immunoblots, mAb IR-2-1 reacted with a band of 67 kDa in all Paramecium species examined . However, no band appeared in the immunoblot of isolated macronuclei of H . obtusa-free P . caudatum and no label was seen in the nuclear matrix of the macronucleus of air-dried P . caudatum . These results suggest that the 67-kDa antigen found in H . obtusa was not imported from the host macronucleus and the same antigen in the host contractile vacuoles and cytoplasm were not derived from the symbiont . These results also showed that an epitope on the decorated spongiome of the Paramecium species is shared by its bacterial symbiont . In contrast to the decorated tubule-specific mAb, DS-1, the antigens for IR-2-1 appeared to be loosely membrane bound as they were lost in paraformaldehyde fixed and acetone permeabilized Paramecium. Neurol Med Chir (Tokyo), 1994 Sep, 34(9), 616 - 9 Subarachnoidal abscess associated with bacterial meningitis in infants--two case reports; Nishizawa S et al.; Two infants, an 11-month-old boy and a 7-month-old girl, presented with subarachnoidal abscess associated with severe bacterial meningitis refractory to intensive managements with antibiotics . Computed tomography (CT) revealed bifrontal extracerebral low-density areas and remarkably enhanced surfaces of the bilateral frontal lobes postcontrast . Surgical exploration disclosed thick pus accumulation in the subarachnoid space which required curettage . The boy developed appropriately for his age, but the girl showed severe psychomotor retardation because of additional complications such as subdural fluid collection and hydrocephalus associated with the subarachnoidal abscess . Appropriate early neurosurgical management of subarachnoidal abscess is essential for satisfactory psychomotor development . Postcontrast CT should be performed to detect the subarachnoidal abscess as early as possible, and extensive craniotomy to remove the subarachnoidal pus accumulation performed to preserve psychomotor development. J Mol Biol, 1994 Aug 19, 241(3), 363 - 77 The modular architecture of bacterial response regulators . Insights into the activation mechanism of the BvgA transactivator of Bordetella pertussis; Boucher PE et al.; Control of virulence factor expression in Bordetella pertussis is mediated by the products of the bvg operon . The BvgS membrane protein responds to certain environmental cues by activating the BvgA protein, which in turn modulates the expression of the target virulence factor genes . The BvgA and BvgS proteins are members of a large family of sensory transduction proteins called the two-component systems . We show that BvgA fusion proteins can activate transcription of a reporter gene containing the bvg promoter in Escherichia coli, and that this activity correlates with its ability to interact specifically with a recognition sequence in cognate promoters . Using homologies between BvgA and other bacterial response regulators as a guide, two BvgA truncation mutants were constructed and their transactivation and DNA-binding capacities were examined . We discovered that (1) DNA-binding activity is localized to the C-terminal half of BvgA, (2) sequence-specific DNA-binding is necessary, but not sufficient for transactivation, and (3) DNA-binding requires the last 20 amino acid residues at its carboxy terminus . A BvgA fusion protein lacking the receiver domain is inactive in transcriptional activation, but retains sequence-specific DNA-binding activity and forms multimeric complexes . We show that BvgA is able to utilize acetyl phosphate as a phosphoryl group donor and the instability of the covalent linkage at extremes of pH is consistent with an acyl phosphate group . Furthermore, the in vitro phosphorylated form of BvgA exhibits an enhanced capacity for binding DNA target sites, while a dephosphorylated form exhibits a limited capacity to bind these sites . We discuss the implications that these observations have on the mechanism by which BvgA is activated to a transcriptionally competent state. Biochem Biophys Res Commun, 1994 Aug 15, 202(3), 1280 - 4 Bacterial expression of a soluble T-cell receptor alpha chain; Stangel M et al.; A bacterial expression system was used to express the complete extracellular domain of a rat T-cell receptor alpha-chain, including variable-, joining and constant regions . By introduction of a His-tail at the C-terminus, a simple and rapid purification protocol utilizing Ni(+)-chelating chromatography was applied, with renaturation of the protein while bound to the matrix . This method produced large amounts of soluble protein, which provide a source for T-cell receptor analysis and further studies on T cell function. J Biol Chem, 1994 Aug 12, 269(32), 20270 - 4 Sugar transport by the bacterial phosphotransferase system . Characterization of the Escherichia coli enzyme I monomer/dimer transition kinetics by fluorescence anisotropy; Chauvin F et al.; Enzyme I of the bacterial phosphoenolpyruvate: glycose phosphotransferase system (PTS) exists in a monomer/dimer (M/D) equilibrium . These two forms are functionally different, and their interconversion may be a means of regulating the PTS . The M/D equilibrium was studied by fluorescence anisotropy of a pyrene derivative (Chauvin, F., Brand, L., and Roseman, S . (1994) J . Biol . Chem . 269, 20263-20269) . In this paper, the kinetics of the transition is investigated . The following apparent rate constants were found for the M/D transition of phospho-Enzyme I in the presence of Mg2+ and PEP at 6 degrees C: k*A = 3.4 x 10(3) M-1 s-1 and k*D = 1.04 x 10(3) s-1 . The association rate is especially slow, 2-3 orders of magnitude slower than the average dimerization rate determined for other proteins . Furthermore, the rate of quaternary structure changes matches that of enzymatic activity changes, as well as that of tertiary structure changes (Chauvin, F., Toptygin, D., Roseman, S., and Brand, L . (1992) Biophys . Chem . 44, 163-173) . Finally, the effect of two ligands is shown; PEP increases the relaxation rate by 3-fold at 23 degrees C, and Mg2+ addition causes a 4-fold increase in the relaxation rate. J Biol Chem, 1994 Aug 12, 269(32), 20263 - 9 Sugar transport by the bacterial phosphotransferase system . Characterization of the Escherichia coli enzyme I monomer/dimer equilibrium by fluorescence anisotropy; Chauvin F et al.; Enzyme I (EI), the first protein of the bacterial phosphotransferase system (PTS), exists in a monomer/dimer (M/D) equilibrium . We have proposed that the two species are functionally different and that their interconversion may regulate sugar transport via the PTS . The C-terminal Cys of Escherichia coli EI was reacted with pyrene maleimide (Han, M . K., Roseman, S., and Brand, L . (1990) J . Biol . Chem . 265, 1985-1995), and the pyrene conjugate used to characterize the M/D equilibrium by fluorescence anisotropy . The properties of unlabeled and pyrene-labeled EI are indistinguishable . Values for the apparent association constant, K'eq, and the steady-state anisotropy of the monomer and the dimer were obtained under a variety of conditions . K'eq increases 23-fold, from 0.45 x 10(5) to 10.7 x 10(5) M-1, as the temperature increases from 6 to 30 degrees C; the association appears to be entropically driven . Under all conditions tested, the K'eq for phospho-EI is 6-12-fold less than for dephospho-EI . For phospho-EI, PEP and Mg2+ induce a 240-fold increase of K'eq when both ligands are present . Based on these data, EI was preincubated under conditions that change K'eq, and the initial activities of the different species were determined at 37 degrees C in a PTS sugar phosphorylation assay with PEP as the phosphoryl donor . The initial rate depends on the M/D ratio; it is maximal when EI is 100% dimer, and zero when EI is 100% monomer . In the latter case, the rate gradually increases in the assay mixture . The results have important implications for how the PTS regulates sugar transport and other physiological phenomena. J Biol Chem, 1994 Aug 5, 269(31), 19701 - 6 Lipid modification of bacterial prolipoprotein . Transfer of diacylglyceryl moiety from phosphatidylglycerol; Sankaran K et al.; The peptide, MKATKLVLGAVILGSTLLAGCSSN, corresponding to the N-terminal 24 amino acids of Braun's prolipoprotein, was used to study the lipid modification of prolipoprotein in Escherichia coli by measuring the rate of incorporation of either {2-3H}glycerol or {9,10-3H}palmitate from the corresponding labeled phosphatidylglycerol into the peptide . Using E . coli strains containing varying levels of prolipoprotein diacylglyceryl modification activities due to mutations in or overexpression of the gene involved in diacylglyceryl modification (lgt), we have shown that the activities based on the peptide assay correlated well with the prolipoprotein-based assay . Further, we have followed the fate of the lipid substrate, phosphatidylglycerol, during the modification reaction and found that lipid modification of prolipoprotein involves the transfer of diacylglyceryl moiety from phosphatidylglycerol to the sulfhydryl group of the cysteine residue with the concomitant formation of sn-glycerol 1-phosphate . This mechanism is contrary to the previously proposed two-step mechanism of an initial glyceryl transferase followed by O-acyl transfer (Chattopadhyay, P.K., and Wu, H.C . (1977) Proc . Natl . Acad . Sci . U . S . A . 74, 5318-5322) . Accordingly, the enzyme that catalyzes this activity has been named phosphatidylglycerol-prolipoprotein diacylglyceryl transferase . The revised pathway for the lipoprotein biogenesis in bacteria consists of three successive reactions catalyzed by prolipoprotein diacylglyceryl transferase, signal peptidase II, and apolipoprotein N-acyltransferase. J Mol Biol, 1994 Aug 5, 241(1), 110 - 24 Filamentous hemagglutinin of Bordetella pertussis . A bacterial adhesin formed as a 50-nm monomeric rigid rod based on a 19-residue repeat motif rich in beta strands and turns; Makhov AM et al.; The filamentous hemagglutinin (FHA) of Bordetella pertussis is an adhesin that binds the bacteria to cells of the respiratory epithelium in whooping-cough infections . Mature FHA is a 220 kDa secretory protein that is highly immunogenic and has been included in acellular vaccines . We have investigated its structure by combining electron microscopy and circular dichroism spectroscopy (CD) with computational analysis of its amino acid sequence . The FHA molecule is 50 nm in length and has the shape of a horseshoe nail: it has a globular head that appears to consist of two domains; a 35 nm-long shaft that averages 4 nm in width, but tapers slightly from the head end; and a small, flexible, tail . Mass measurements by scanning transmission electron microscopy establish that FHA is a monomer . Its sequence contains two regions of tandem 19-residue pseudo-repeats: the first, of 38 cycles, starts at residue 344; the second, of 13 cycles, starts at residue 1440 . The repeat motifs are predicted to consist of short beta-strands separated by beta-turns, and secondary structure measurements by CD support this prediction . We propose a hairpin model for FHA in which the head is composed of the terminal domains; the shaft consists mainly of the repeat regions conformed as amphipathic, hyper-elongated beta-sheets, with their hydrophobic faces apposed; and the tail is composed of the intervening sequence . Further support for the model was obtained by immuno-labeling electron microscopy . The 19-residue repeats of FHA have features in common with the leucine-rich repeats (LRRs) that are present in many eukaryotic proteins, including some adhesion factors . The model is also compared with the two other classes of filamentous proteins that are rich in beta-structure, i.e . viral adhesins and two beta-helical secretory proteins . Our proposed structure implies how the functionally important adhesion sites and epitopes of FHA are distributed: its tripeptide (RGD) integrin-binding site is assigned to the tail; the putative hemagglutination site forms part of the head; and two classes of immunodominant epitopes are assigned to opposite ends of the molecule . Possible mechanisms are discussed for two modes of FHA-mediated adhesion. Acta Obstet Gynecol Scand, 1994 Aug, 73(7), 586 - 8 Bacterial vaginosis and cervical intraepithelial neoplasia; Platz-Christensen JJ et al.; METHODS . In an attempt to investigate an association between the finding of clue cells in Papanicolaou-stained (PAP) smears and cervical intraepithelial neoplasia (CIN), a total of 6150 smears from 1976 were re-investigated . RESULTS . Clue cells representing bacterial vaginosis were present in 10% of the PAP-smears . CIN II and III alone, as well as all CIN cases, were more common in women with bacterial vaginosis (p < 0.001) . Histologically CIN III/carcinoma in situ was found in nine patients with and in 16 patients without bacterial vaginosis . The relative risk of having CIN III/carcinoma in situ if the women had bacterial vaginosis was 5.0 with 95% confidence interval of 2.2-11.6 . CONCLUSIONS . The possibility exists that bacterial vaginosis is in some way associated with the development of cervical intraepithelial neoplasia, i.e . as a cofactor to human papilloma virus . Therefore, bacterial vaginosis must be taken in consideration in future studies on CIN. Am J Obstet Gynecol, 1994 Aug, 171(2), 345 - 7; discussion 348-9 Effect of metronidazole in patients with preterm birth in preceding pregnancy and bacterial vaginosis: a placebo-controlled, double-blind study; Morales WJ et al.; OBJECTIVE: Our purpose was to determine whether treatment of bacterial vaginosis with metronidazole in patients with preterm delivery in the penultimate pregnancy from preterm labor or premature rupture of membranes reduces the risk of subsequent preterm birth . STUDY DESIGN: From January 1989 to June 1992 patients with a singleton gestation between 13 and 20 weeks and a history of preterm birth in the preceding pregnancy from either idiopathic preterm labor or premature rupture of membranes were screened for bacterial vaginosis . Those with a positive screen were randomized to receive 250 mg of metronidazole three times a day for 7 days or placebo in a double-blind design . Data were analyzed with Student t and chi 2 tests, and differences considered significant at p < 0.05 . RESULTS: Of 94 eligible patients, 80 were enrolled and completed the study, of which 44 received metronidazole . Both groups were comparable in number of entry variables . Compared with the placebo group, patients in the metronidazole group had significantly fewer hospital admissions for preterm labor, 12 (27%) versus 28 (78%); preterm births, eight (18%) versus 16 (39%); births of infants weighing < 2500 gm, six (14%) versus 12 (33%); and premature rupture of membranes, two (5%) versus 12 (33%) . CONCLUSION: Treatment of bacterial vaginosis with metronidazole was effective in reducing preterm births in patients with a history of prematurity in the preceding pregnancy. Ann Surg, 1994 Aug, 220(2), 193 - 8 Time course of bacterial infection of the pancreas and its relation to disease severity in a rodent model of acute necrotizing pancreatitis; Foitzik T et al.; BACKGROUND: Bacterial infection of pancreatic necrosis is thought to be a major determinant of outcome in acute necrotizing pancreatitis . The determinants and possibilities for prophylaxis are unknown and difficult to study in humans . OBJECTIVE: The time course of bacterial infection of the pancreas in a rodent model of acute necrotizing pancreatitis was characterized . The authors ascertained if there is a correlation with the degree of necrosis . METHODS: Acute pancreatitis (AP) of graded severity was induced under sterile conditions by an intravenous infusion of cerulein (5 micrograms/kg/hr) for 6 hours (mild AP), or a combination of intravenous cerulein with an intraductal infusion of 10-mM glycodeoxycholic acid (0.2 mL for 2 min for moderate AP, 0.5 mL for 10 min for severe AP) . Sham-operated animals (intravenous and intraductal NaCl 0.9%) served as controls . Ninety-six hours after induction, animals were killed for quantitative bacterial examination and histologic scoring of necrosis . In addition, groups of animals with severe AP were investigated at 12, 24, 48, 96, and 144 hours . RESULTS: No significant pancreatic necrosis was found in control animals (0.3 +/- 0.1) or animals with mild AP (0.6 +/- 0.1) killed at 96 hours . Necrosis scores were 1.1 +/- 0.2 for animals with moderate AP and 1.9 +/- 0.2 for animals with severe AP . Control animals did not develop significant bacterial infection of the pancreas (> or = 10(3) CFU/g) . At 96 hours, the prevalence of infection was 37.5% in animals with mild AP and 50% in animals with moderate AP . In animals with severe AP, infection of the pancreas increased from 33% in the first 24 hours to 75% between 48 and 96 hours (p < 0.05) . The bacterial counts and the number of different species increased with time and was maximal (> 10(11) CFU/g) at 96 hours . CONCLUSION: Bacterial infection of the pancreas in rodent AP increases during the first several days, and its likelihood correlates with the severity of the disease . This model, which closely mimics the features of human acute pancreatitis, provides a unique opportunity to study the pathogenesis of infected necrosis and test therapeutic strategies. Am J Gastroenterol, 1994 Aug, 89(8), 1238 - 40 Ascites and secondary bacterial peritonitis associated with small bowel obstruction; Scheider DM et al.; Wide albumin gradient (transudative) ascites is usually due to liver disease but may also result from many other disorders, including heart failure, hepatic infiltration by tumor, hepatic vein thrombosis, and veno-occlusive disease . It has not been linked with small bowel obstruction . Narrow albumin gradient (exudative) ascites, usually due to peritoneal carcinoma or inflammation, has been noted in cases of necrotic or perforated bowel, but simple small bowel obstruction has not previously been appreciated as a possible cause for ascites . We report a patient who developed wide albumin gradient ascites and secondary bacterial peritonitis in association with small bowel obstruction . The small bowel obstruction, ascites, and peritonitis resolved with lysis of a single abdominal adhesion. Am J Gastroenterol, 1994 Aug, 89(8), 1211 - 8 Helicobacter pylori infection and serum pepsinogen A, pepsinogen C, and gastrin in gastritis and peptic ulcer: significance of inflammation and effect of bacterial eradication; Wagner S et al.; OBJECTIVES: To study the relationship between Helicobacter pylori infection, gastric inflammatory scores, and fasting gastrin and pepsinogen A and C concentrations, and to evaluate the effect of treatment on these parameters . METHODS: Gastrin and pepsinogen A and C concentrations were measured in 36 patients with gastritis, 10 gastric ulcer patients, 12 duodenal ulcer patients, and in 15 subjects with normal gastric mucosa, by standard radioimmunoassay techniques . Fifteen patients with H . pylori infection underwent triple therapy (bismuth subsalicylate, amoxicillin, metronidazole) and were reassessed 1 month later . RESULTS: Fasting gastrin and pepsinogen A and C concentrations were significantly higher in H . pylori-positive gastritis and peptic ulcer patients than in subjects with normal mucosa and in patients with H . pylori-negative gastritis . There was a significant correlation between inflammatory scores and serum gastrin (r = 0.45, p < 0.0001), and pepsinogen A (r = 0.33, p < 0.006) and pepsinogen C (r = 0.55, p < 0.0001) concentrations . Neither sex nor age affected basal gastrin and pepsinogen concentrations . Eradication of H . pylori infection was successful in 12 patients and resulted in a significant fall in serum gastrin and in pepsinogen A and C concentrations, and in a concomitant improvement of the inflammatory scores . Serum peptide levels and gastritis scores were unchanged in those patients in whom H . pylori infection persisted . CONCLUSIONS: These findings suggest that hypergastrinemia and hyperpepsinogenemia are secondary to H . pylori infection and are related to mucosal inflammation. J Pediatr, 1994 Aug, 125(2), 235 - 8 Clinical usefulness of cerebrospinal fluid bacterial antigen studies; Maxson S et al.; A retrospective chart review was performed to evaluate the effect that positive results of cerebrospinal fluid bacterial antigen tests had on the care of patients with presumed bacterial meningitis . Of 901 tests ordered, costing $26,000 per year, 29 showed positive results--and only four of these affected patient care . By using cerebrospinal fluid bacterial antigen testing only when another test does not identify an organism, or in an attempt to determine central nervous system infection late in therapy for presumed sepsis, one can greatly reduce costs with no detrimental effect on patients. J Surg Res, 1994 Aug, 57(2), 238 - 45 Obstructive jaundice impairs reticuloendothelial function and promotes bacterial translocation in the rat; Ding JW et al.; Septic complications and renal insufficiency following biliary tract surgery are frequently seen in patients with obstructive jaundice . The precise mechanisms for understanding the susceptibility of the jaundiced patients to sepsis are, however, not clear . The present study aimed at investigating the influence of biliary obstruction on the reticuloendothelial function and bacterial translocation at various time intervals in the rat . Reticuloendothelial system (RES) function, as evaluated by measuring blood clearance of intravenously injected 125I-labeled Escherichia coli, and bacterial translocation were studied 3 days and 1, 2, and 3 weeks following either sham operation or common bile duct ligation (CBDL) and transection in the rat . RES function was significantly impaired and renal uptake of radiolabeled E . coli was significantly higher in jaundiced animals from Day 3 and on after CBDL (P < 0.01) concomitant with elevation of plasma levels of bilirubin and liver enzymes (P < 0.001) compared with their corresponding controls . The incidence of bacterial translocation 3 days and 1 and 2 weeks after biliary obstruction significantly increased (P < 0.05) . We conclude that RES phagocytic function is impaired and the incidence of bacterial translocation is increased in jaundiced rats . These findings might contribute to explain the high susceptibility of postoperative septic complications and renal dysfunction in patients with obstructive jaundice. Clin Otolaryngol, 1994 Aug, 19(4), 310 - 3 Bacterial invasion of the tonsillar tissues in acute pharyngotonsillitis and in the adenoid: a preliminary study; Ebenfelt A et al.; In order to understand the immunological and inflammatory processes in which the tonsils are involved it is necessary to know the spatial relation between bacteria and the tissues . In this study four adenoids and four palatine tonsils obtained at elective surgery and four palatine tonsils obtained during a chaud surgery for quinsy were examined histologically . Acridine orange and fluorescence microscopy were used to identify bacteria in tonsillar tissue . The adenoids were also stained with haematoxillin-eosin . Bacteria were in every case seen on the surfaces and in the crypts of the tonsils and adenoids . In the tissues, however, bacteria were never seen irrespective of whether the tonsils were obtained during an acute infection or not . We conclude that bacterial invasion in tonsillar tissue is neither a prerequisite for a clinically manifest acute bacterial pharyngotonsillitis nor a common feature in tonsils clinically recognized as non-infected. Comput Methods Programs Biomed, 1994 Aug, 44(2), 61 - 7 Shading correction and calibration in bacterial fluorescence measurement by image processing system; Wilkinson MH; An image processing system with applications in bacterial (immuno-)fluorescence measurement has been developed . To reach quantitative results, correction for non-uniformities in system sensitivity, both as a function of time (calibration for drifts) and as a function of image coordinates (shading correction), is essential . Both problems can be handled simultaneously by acquiring images of a uniformly fluorescent, solid standard as a reference image . To measure bacterial fluorescence, the average fluorescence intensity of isolated areas of interest (the bacteria) is computed, and corrected using the reference images . Two shading correction methods are theoretically and experimentally compared: direct averaging in the corrected image, and (weighted and unweighted) averaging using the raw image and a separate shading image to determine the weights and correct for shading during the averaging . The latter method proved computationally 3.5-6.5 times faster on average and reduced propagation of truncation errors during computation, resulting in 40% less noise, for 8-bits/pixel images. Am J Vet Res, 1994 Aug, 55(8), 1127 - 38 Effect of bacterial lipopolysaccharides on sulfated glycosaminoglycan metabolism and prostaglandin E2 synthesis in equine cartilage explant cultures; MacDonald MH et al.; The metabolic responses of equine articular cartilage to incubation with bacterial lipopolysaccharide (LPS) were studied, using explant cultures of articular cartilage obtained from the metatarsophalangeal joints of 15 horses, age of which ranged from 3 months to 20 years . For comparison, explants were also established from the metatarsophalangeal joints of 3 calves . Explants were cultured for 3 days in medium containing various concentrations of LPS from 0 (control) to 100 micrograms/ml . Glycosaminoglycan (GAG) released during the 3-day incubation was determined by a spectrophotometric assay, using the dye 1,9-dimethylmethylene blue . Newly synthesized GAG content was assayed by measuring {35S}sulfate incorporation during a 3-hour pulse labeling period . In addition, prostaglandin E2 (PGE2) synthesis was quantified, using a {3H}PGE2 radioimmunoassay kit and magnetic separation . Finally, explants from 3 animals were used to evaluate the effect of supplementing culture medium with 5% serum on the response of explants to LPS, and explants from 1 horse were used to compare responses to stimulation with LPS derived from 2 bacterial sources . Equine explants cultured with bacterial LPS had a dose-dependent decrease in synthesis and increase in release of GAG, and these responses were significantly (P < 0.0001) greater in explants from younger horses . In addition, equine explants had a significant (P = 0.0001) dose-dependent increase in concentration of PGE2 released into the culture medium in response to incubation with LPS . Comparison of data for GAG synthesis from equine and bovine explants revealed a significant (P = 0.025) difference in responsiveness to LPS between the 2 species . Equine explants tended to have a greater suppression of GAG synthesis in response to incubation with increasing concentrations of LPS than did age-corrected bovine samples . However, similar analysis of data on GAG release did not indicate any difference in sensitivity between the 2 species for this response . There was no evidence that the presence or absence of serum supplementation or the use of LPS derived from different bacterial sources made a significant difference in the response of explants to incubation with LPS. Anal Biochem, 1994 Aug 1, 220(2), 410 - 4 Bioluminescent assay of bacterial intracellular AMP, ADP, and ATP with the use of a coimmobilized three-enzyme reagent (adenylate kinase, pyruvate kinase, and firefly luciferase); Brovko LYu et al.; A three-enzyme coimmobilized system (firefly luciferase, pyruvate kinase, and adenylate kinase) was constructed for the bioluminescent assay of ATP, ADP, and AMP in bacterial cell extracts . Data for the reproducibility and sensitivity of the proposed method are presented . Detection limits were 1.5 pmol of ADP and 15 pmol of AMP in the sample . With this system, changes in adenine nucleotide concentrations in bacterial cells were measured during the actions exerted by external chemical and physical sources, such as additives to nutrient media and low-power He-Ne laser irradiation. J Pediatr Surg, 1994 Aug, 29(8), 1059 - 63; discussion 1063-4 Breast milk protects the neonate from bacterial translocation; Go LL et al.; Clinical and experimental evidence suggests that breast milk enhances the neonatal gut barrier . Using bacterial translocation (BT) as a measure of gut barrier function, a series of experiments was designed to explore the relationship between the neonatal gut barrier and breast milk as well as the factors associated with the feeding of breast milk . Full-term newborn rabbits were assigned to one of four groups: formula-fed (group I), fed with colostrum plus formula (group II), breast-fed with breast milk (group III), and fed with colostrum plus stored breast milk (group IV) . At 7 days of age, body weights were obtained, the rabbits were killed, and the mesenteric lymph nodes (MLN), liver, and spleen were quantitatively cultured for translocating bacteria . The cecum was cultured for aerobic and anaerobic enteric organisms . Distal ileal tissues were examined by light and transmission electron microscopy and compared among groups . The viability of cells in the stored, frozen breast milk was assessed by Trypan blue staining . Group I rabbits had significantly lower mean body weights compared with the other groups . The animals breast-fed breast milk had no BT to the MLN or liver and had a 9% incidence of BT to the spleen . There was no difference between BT in groups III and IV . The stored breast milk contained no viable cells . The incidence of BT to all three areas was significantly lower than in groups I and II . The animals fed with formula alone had the highest incidence of BT to the MLN (88%), liver (60%), and spleen (32%) . BT in this group was significantly higher compared with groups III and IV.(ABSTRACT TRUNCATED AT 250 WORDS) Nippon Kyobu Geka Gakkai Zasshi, 1994 Aug, 42(8), 1231 - 4 {Mitral valve replacement secondary to resection of mycotic cerebral aneurysm in acute phase of bacterial endocarditis--a case report}; Fujii Y et al.; We report a case of mitral valve replacement after ruptured mycotic aneurysm resection in acute phase of bacterial endocarditis . We have experienced a 68-year-old man with vegetation at the anterior leaflet of mitral valve and multiple systemic embolization . He underwent aneurysmectomy of ruptured mycotic cerebral aneurysm and embolectomy of left femoral artery eight days after subarachnoid hemorrhage . Mitral valve was replaced three days after successfully . If there was no heart failure preoperatively, valve replacement operation is recommended in acute phase of infected endocarditis or few days after cerebral aneurysmectomy. Ginecol Obstet Mex, 1994 Aug, 62, 226 - 34 {New alternatives in the treatment of bacterial vaginosis}; Luis Arredondo J et al.; Efficiency and security clindamycin vaginal cream (2%) were compared to oral metronidazole's for the treatment of 184 women with symptomatic bacterial vaginosis in a multicentric, randomized, double-blind, controlled study . The treatment was of 7 days duration, using placebo capsules for the clindamycin group and placebo cream for the metronidazole group . Patients were observed during a follow up (4-13 and 20-43 days after completion of therapy) . Global results of this treatment indicated that clindamycin vaginal cream offers a similar efficiency than oral metronidazole . Improvement or total healing was 87% for clindamycin and 79% for metronidazole, with no significant differences (p > 0.22) . No relapses were observed in the clindamycin group, and 7% in the metronidazole group . The clindamycin group had a failure rate of 3% compared to 15% in the oral metronidazole group . Both drugs were well tolerated . Side effects more frequently reported were vulvovaginal irritation and cervicitis/vaginitis . The only side effect that could have been classified as serious was a generalized rash in a patient receiving metronidazole . It was concluded that clindamycin vaginal cream (2%) is an efficient and secure alternative to oral metronidazole for the treatment of bacterial vaginosis being the elective therapy for pregnant women in their first gestational trimester. Eur J Pediatr, 1994 Aug, 153(8), 565 - 8 The variation of carnitine content in human blood cells during disease--a study in bacterial infection and inflammatory bowel disease; Demirkol M et al.; Carnitine in erythrocytes and leucocytes represents a small but essential part of the cellular carnitine pool . It was the objective of this study to document the changes of blood cell carnitine concentrations in disease entities with an enhanced cellular metabolism during acute and chronic inflammation . The plasma, erythrocyte, lymphocyte, granulocyte and thrombocyte carnitine concentrations were determined in 23 patients (11.0 +/- 8.8 years) with bacterial infections and nine patients (17.5 +/- 2.4 years) with Crohn disease and compared to 20 healthy controls (27.0 +/- 10.6 years) . In patients with bacterial infections the granulocyte carnitine concentrations (126.4 +/- 73.5 nmoles/10(6) cells) were higher (P < 0.001) than in controls (37.9 +/- 22.8 nmoles/10(6) cells) . In patients with Crohn disease the lymphocyte carnitine concentrations (169.4 +/- 108.2 nmoles/10(6) cells) were increased (P < 0.001) when compared to controls (48.1 +/- 18.3 nmoles/10(6) cells) . The plasma carnitine concentrations were decreased (P < 0.05) in both patient groups, whereas they were increased (P < 0.05) in the patients' erythrocytes . The carnitine concentrations in thrombocytes did not differ significantly within the individual groups . CONCLUSION: Lymphocyte and granulocyte carnitine concentrations may reflect their enhanced metabolic state during either immunoglobulin formation or phagocytosis. Zhonghua Yi Xue Za Zhi (Taipei), 1994 Aug, 54(2), 87 - 92 Serum and ascitic concentration of C3, C4 and protein in cirrhotic patients with spontaneous bacterial peritonitis; Chen SM et al.; BACKGROUND . Lower concentration of ascitic or serum complement (C3, C4) or protein has been reported to participate in the development of spontaneous bacterial peritonitis (SBP) . In Taiwan, the etiology of hepatic cirrhosis is mainly post-hepatic and SBP is the common complication . This study aims to determine the role of protein and complements in the pathogenesis of SBP . METHODS . 119 cirrhotic patients were divided into two groups, 30 SBP and 89 non-SBP . The concentrations of ascitic and serum complement and protein were measured for comparison . RESULTS . The ascitic and serum C3, C4 and protein levels were significantly lower (P < 0.05) in patients with SBP than in non-SBP patients . No significant differences were noted in the ascites/serum ratio of C3, C4 and protein in patient with or without SBP . CONCLUSIONS . Low levels of ascitic and serum protein and complements, C3 and C4, may be prone to develop SBP in our patients mostly with post-hepatitic cirrhosis. Curr Opin Immunol, 1994 Aug, 6(4), 518 - 25 Bacterial and protozoal infections in genetically disrupted mice; Kaufmann SH; Immune gene knock-out mice have provided challenging new opportunities for scientists interested in infection and immunity . This novel experimental approach has made it possible to gain deeper insights into the mechanisms that are operative in defence against, and the pathology of, infectious disease . It has allowed the identification not only of vital immune elements, but also of compensatory mechanisms and of novel cells that are still uncharacterized or of uncertain function. J Trop Pediatr, 1994 Aug, 40(4), 231 - 4 Localized extracranial infections in children with acute bacterial meningitis; Akpede GO; Twenty-four of 66 (363.6 +/- 116/1000) post-neonatal children with acute bacterial meningitis treated between August 1991 and November 1992 at the Emergency Paediatric Unit, University of Maiduguri Teaching Hospital, Nigeria, had co-existing localized extracranial infections (LEI) . Nineteen had bronchopneumonia (of which 14 were in combination with other infections), two had dysentery and one each acute otitis media, orbital cellulitis, and purulent conjunctivitis . LEI were significantly more frequent in children < or = 2 years, and in children with delayed presentation and delayed diagnosis after presentation . Mortality rate was higher in children with LEI (500 +/- 200/1000) than in those without (102.6 +/- 95.2/1000) (P < 0.001) . There was no significant relationship between the incidence of LEI and the causative organisms of meningitis or nutritional status of children with meningitis . We conclude that co-existing LEI are not infrequent in children with acute bacterial meningitis; they contribute to delayed diagnosis and are associated with a poorer outcome. FEMS Microbiol Rev, 1994 Aug, 14(4), 375 - 9 Mineralization and responses of bacterial spores to heat and oxidative agents; Marquis RE et al.; Mineralization of bacterial spores with Ca2+ and a variety of other mineral cations enhances resistance to heat damage . Part of the enhancement is associated with increased dehydration of the mineralized protoplast or spore core, while part is independent of dehydration and effective for resistance even to dry heat . Spore mineralization was found also to enhance resistance to oxidative damage caused by agents such as tertiary butyl hydroperoxide or H2O2 . In contrast, mineral cations in the environment increased oxidative damage, presumably by catalyzing radical formation . Metal ion chelators such as o-phenanthroline protected spores against such damage. Mol Biotechnol, 1994 Aug, 2(1), 29 - 44 Bacterial genome mapping by two-dimensional pulsed-field gel electrophoresis (2D-PFGE); Bautsch W; Macrorestriction mapping is often the first step toward a thorough physical and genetic characterization of a bacterial genome . The problem of deducing the order of partially or completely digested macrorestriction fragments to yield a physical genome map may readily be solved by applying two-dimensional pulsed-field gel electrophoresis (2D-PFGE) techniques . These powerful methods are quick and technically easy to perform; specifically, they are independent of DNA probes and should therefore be applicable to any bacterial species irrespective of its prior genetic characterization . In this article, detailed step-by-step protocols are given to set up, run, and evaluate 2D pulsed-field gels . Two basic methods are described: partial/complete 2D gels of one restriction enzyme and complete/complete 2D gels of two different restriction enzymes . Other topics include preparation of bacterial genomic DNA, screening for suitable rare-cutting restriction enzymes and determination of optimal running conditions . Accompanied by many notes, these protocols are meant to offer the novice a sound and rapid access to these important methods. Zh Mikrobiol Epidemiol Immunobiol, 1994 Aug-Sep, Suppl 1, 83 - 7 {Bacterial pathogenicity factors: their functions in the environment}; Litvin VIu et al.; On the basis of literature data and the results of experimental studies the concept of the universal character of the main pathogenicity factors of bacteria (adhesion, enzymes, toxins, resistance to phagocytosis) as the adaptive mechanisms of bacterial populations under different ecological conditions, including their existence in the environment, has been put forward . Resistance of bacteria to phagocytosis as the mechanism of their protection from predators (protozoa, etc.), necessary for the existence bacteria in the ecological systems of water and soil, is considered in detail. Acta Paediatr Suppl, 1994 Aug, 400, 46 - 50 The opportunistic and bacterial infections associated with pediatric human immunodeficiency virus disease; Nicholas SW; Opportunistic and bacterial infections remain the leading causes of death of Human Immunodeficiency Virus-infected children, despite recent advances in the diagnosis of HIV infection during early infancy; antiretroviral therapies; advances in the treatment of some infections; an improved understanding of the cellular immune systems during early childhood; and new strategies for the prevention of some infections . However, these advances appear to be changing the natural history of pediatric HIV infection, resulting in an improved and longer life for infected children . This article briefly reviews the epidemiology, predictors, and treatments of the most common infections associated with pediatric HIV disease, including Pneumocystis carinii pneumonia, recurrent bacterial infections, candidiasis, herpes group viruses, mycobacterial disease and cryptosporidiosis. Curr Opin Obstet Gynecol, 1994 Aug, 6(4), 389 - 91 Vaginitis including bacterial vaginosis; Eschenbach DA; Bacterial vaginosis is a common lower genital tract infection . Women with bacterial vaginosis have 100-1000 times more virulent bacterial per ml of vaginal flora than women without this infection . This tremendous increase in the concentration of bacteria has been recently associated with postpartum and posthysterectomy infection and preterm delivery. Glia, 1994 Aug, 11(4), 336 - 44 Bacterial endotoxin induces {Ca2+}i transients and changes the organization of actin in microglia; Bader MF et al.; We have employed amoeboid microglia purified from primary cultures of neonatal rat brain to examine the effect of bacterial lipopolysaccharide (LPS), a potent activator of immune cells, on intracellular calcium concentration ({Ca2+}i) in brain macrophages . In single brain macrophages loaded with indo 1, pulse administration of LPS elicited a rapid and transient increase in {Ca2+}i . From a total of 70 cells examined, all responded to LPS with a similar {Ca2+}i transient, indicating a good homogeneity of the cell population with regard to the LPS response . It was concluded that the rise of cytosolic {Ca2+}i originated from intracellular stores because the response to LPS occurred similarly in the presence or in the absence of extracellular Ca2+ . A second administration of LPS to the same cells resulted in a second but reduced {Ca2+}i transient . In contrast to the first response to LPS, this second response was totally dependent on the presence of Ca2+ in the extracellular medium . The first response to LPS was strongly inhibited by ruthenium red and could be suppressed in a reversible manner by preincubating the cells with caffeine in the absence of Ca2+ in the extracellular medium . These results indicate that caffeine-sensitive intracellular Ca2+ stores may be the major source of Ca2+ in the response of brain macrophages to LPS . The possible release of Ca2+ from phosphatidylinositol(3,4,5)-trisphosphate (IP3)-sensitive stores in brain macrophages was also evaluated by stimulating cells with the IP3-mobilizing agonist histamine . Brain macrophages were heterogeneous with regard to the histamine response since histamine induced a {Ca2+}i rise in only 30% of cells examined.(ABSTRACT TRUNCATED AT 250 WORDS) Proc Natl Acad Sci U S A, 1994 Jul 19, 91(15), 6808 - 14 Dynamics of adaptation and diversification: a 10,000-generation experiment with bacterial populations; Lenski RE et al.; We followed evolutionary change in 12 populations of Escherichia coli propagated for 10,000 generations in identical environments . Both morphology (cell size) and fitness (measured in competition with the ancestor) evolved rapidly for the first 2000 generations or so after the populations were introduced into the experimental environment, but both were nearly static for the last 5000 generations . Although evolving in identical environments, the replicate populations diverged significantly from one another in both morphology and mean fitness . The divergence in mean fitness was sustained and implies that the populations have approached different fitness peaks of unequal height in the adaptive landscape . Although the experimental time scale and environment were microevolutionary in scope, our experiments were designed to address questions concerning the origin as well as the fate of genetic and phenotypic novelties, the repeatability of adaptation, the diversification of lineages, and thus the causes and consequences of the uniqueness of evolutionary history . In fact, we observed several hallmarks of macroevolutionary dynamics, including periods of rapid evolution and stasis, altered functional relationships between traits, and concordance of anagenetic and cladogenetic trends . Our results support a Wrightian interpretation, in which chance events (mutation and drift) play an important role in adaptive evolution, as do the complex genetic interactions that underlie the structure of organisms. Nucleic Acids Res, 1994 Jul 11, 22(13), 2634 - 6 Sequence similarity of putative transposases links the maize Mutator autonomous element and a group of bacterial insertion sequences; Eisen JA et al.; The Mutator transposable element system of maize is the most active transposable element system characterized in higher plants . While Mutator has been used to generate and tag thousands of new maize mutants, the mechanism and regulation of its transposition are poorly understood . The Mutator autonomous element, MuDR, encodes two proteins: MURA and MURB . We have detected an amino acid sequence motif shared by MURA and the putative transposases of a group of bacterial insertion sequences . Based on this similarity we believe that MURA is the transposase of the Mutator system . In addition we have detected two rice cDNAs in genbank with extensive similarity to MURA . This sequence similarity suggests that a Mutator-like element is present in rice . We believe that Mutator, a group of bacterial insertion sequences, and an uncharacterized rice transposon represent members of a family of transposable elements. Microbiology, 1994 Jul, 140 ( Pt 7), 1575 - 83 Characterization of attached bacterial populations in deep granitic groundwater from the Stripa research mine by 16S rRNA gene sequencing and scanning electron microscopy; Ekendahl S et al.; This paper presents the molecular characterization of attached bacterial populations growing in slowly flowing artesian groundwater from deep crystalline bed-rock of the Stripa mine, south central Sweden . Bacteria grew on glass slides in laminar flow reactors connected to the anoxic groundwater flowing up through tubing from two levels of a borehole, 812-820 m and 970-1240 m . The glass slides were collected, the bacterial DNA was extracted and the 16S rRNA genes were amplified by PCR using primers matching universally conserved positions 519-536 and 1392-1405 . The resulting PCR fragments were subsequently cloned and sequenced . The sequences were compared with each other and with 16S rRNA gene sequences in the EMBL database . Three major groups of bacteria were found . Signature bases placed the clones in the appropriate systematic groups . All belonged to the proteobacterial groups beta and gamma . One group was found only at the 812-820 m level, where it constituted 63% of the sequenced clones, whereas the second group existed almost exclusively at the 970-1240 m level, where it constituted 83% of the sequenced clones . The third group was equally distributed between the levels . A few other bacteria were also found . None of the 16S rRNA genes from the dominant bacteria showed more than 88% similarity to any of the others, and none of them resembled anything in the database by more than 96% . Temperature did not seem to have any effect on species composition at the deeper level . SEM images showed rods appearing in microcolonies.(ABSTRACT TRUNCATED AT 250 WORDS) Microbiology, 1994 Jul, 140 ( Pt 7), 1565 - 73 Carbon transformations by attached bacterial populations in granitic groundwater from deep crystalline bed-rock of the Stripa research mine; Ekendahl S et al.; This paper presents and compares the assimilation rates of CO2 and lactate, and the lactate respiration rates, of attached bacterial populations growing in slowly flowing groundwater (1-3 mm s-1) from deep crystalline bed-rock of the Stripa research mine, Sweden . The bacteria studied grew in anoxic, high-pH (9-10) and low-redox artesian groundwater flowing up through tubing from two levels of a borehole designated V2, 812-820 m and 970-1240 m below ground . Bacteria were allowed to attach to and grow on sterile glass microscope slides in laminar-flow reactors connected to the flowing groundwater . Total numbers of bacteria were counted by acridine orange direct counts . The bacteria grew slowly, with doubling times of 34 d at 10 degrees C for the 812-820 m population, 23 d for the 970-1240 m population at 10 degrees C and 16 d for this population at 20 degrees C . Numbers of attached bacteria reached between 10(6) and 10(7) bacteria cm-2 . The populations at the two levels of the borehole were different in physiology as well as in phylogeny and reflected the heterogeneity between the sampling levels . The earlier proposed presence of sulphate-reducing bacteria could not be confirmed . This is discussed in relation to results from 16S rRNA gene sequencing studies . The CO2 assimilation rates (as mol CO2 cm-2 h-1, using liquid scintillation techniques) increased with depth and temperature.(ABSTRACT TRUNCATED AT 250 WORDS) Curr Opin Rheumatol, 1994 Jul, 6(4), 394 - 400 Bacterial arthritis; Goldenberg DL; Acute bacterial arthritis continues to be a common occurrence, and despite improved treatment regimens, the prognosis has not improved significantly over the past 20 years . During the past year, the emergence of unique predisposing factors for bacterial arthritis, including prosthetic joints and HIV infection, were emphasized . The long-standing debate regarding optimal drainage techniques has continued with the introduction of a new drainage procedure, tidal irrigation, in the treatment of septic knees. Clin Exp Immunol, 1994 Jul, 97(1), 33 - 8 Susceptibility to adjuvant arthritis: relative importance of adrenal activity and bacterial flora; van de Langerijt AG et al.; Previous studies on the regulation of bacterial-induced arthritis in rats have focused on endocrine aspects as well as differences in T cell immunity against bacterial epitopes . We analysed the role of both adrenal activity and bacterial flora in determining susceptibility to bacterial-induced arthritis . Outbred Wistar rats show a low incidence of adjuvant arthritis . Moderate sensitivity to adjuvant arthritis was found in a selected, stress-resistant line of the Wistar rat, whereas no arthritis was found in a stress-susceptible Wistar line . Plasma corticosterone responses after IL-1 alpha exposure were, however, identical in these two lines, excluding a direct correlation between susceptibility and corticosterone levels . In line with previous findings in germ-free (GF) F344 rats, GF Wistars also appeared highly susceptible to arthritis . We further analysed the corticosterone responses in GF and conventional (CV) rats . Administration of IL-1 alpha induced identical corticosterone responses in both CV and GF F344 rats . In addition, plasma corticosterone levels were measured around the time of onset of arthritis . Whereas no rise was seen in the arthritis-resistant CV rats, a significant increase was observed from day 14 in GF rats, at the moment of onset of arthritis . Although this corticosterone response was insufficient to prevent arthritis, it may have ameliorated disease expression in the GF F344 rats . Our data indicate that the bacterial flora, and therefore T cell tolerance, is of prime importance in determining susceptibility, whereas the activity of the hypothalamus-pituitary-adrenal (HPA) axis may modulate disease severity. J Biol Chem, 1994 Jul 1, 269(26), 17606 - 10 Bacterial lipopolysaccharide has structural similarity to ceramide and stimulates ceramide-activated protein kinase in myeloid cells; Joseph CK et al.; Bacterial lipopolysaccharide (LPS), tumor necrosis factor (TNF)-alpha and interleukin-1 beta (IL-1 beta) stimulate similar cellular responses . TNF-alpha and IL-1 beta are known to initiate signaling through a pathway involving hydrolysis of sphingomyelin to ceramide (Kolesnick, R . N., and Golde, D . W . (1994) Cell 77, 325-328) . In this system, ceramide acts as a second messenger stimulating a ceramide-activated serine/threonine protein kinase . The present studies demonstrate that LPS, like TNF and IL-1, stimulates ceramide-activated protein kinase activity in human leukemia (HL-60) cells and in freshly isolated human neutrophils . Lipid A, the biologically active core of LPS, enhanced kinase activity in a time- and concentration-dependent manner . As little as 10 nM lipid A was effective, and a maximal effect occurred with 500 nM lipid A, increasing kinase activity 5-fold . Native LPS similarly induced kinase activation . This effect of LPS was markedly enhanced by LPS binding protein and required the LPS receptor CD14 . In contrast to TNF and IL-1, LPS did not cause sphingomyelin hydrolysis and thus stimulates ceramide-activated protein kinase without generating ceramide . Molecular modeling showed strong structural similarity between ceramide and a region of lipid A . Based on these observations, we propose that LPS stimulates cells by mimicking the second messenger function of ceramide. Gastroenterology, 1994 Jul, 107(1), 231 - 5 Lipolytic activity of bacterial lipase survives better than that of porcine lipase in human gastric and duodenal content; Raimondo M et al.; BACKGROUND/AIMS: Treatment of pancreatic steatorrhea with porcine lipase is unsatisfactory because it is rapidly denatured by acidic intraluminal conditions . The aim of this study was to determine if bacterial lipase is resistant to acid denaturation and is active in the presence of bile acids by comparing its stability with that of porcine lipase in gastric and duodenal juice obtained from six patients undergoing a cholecystokinin octapeptide pancreatic function test . METHODS: After inactivating native lipolytic activity, both juices were altered to simulate fasting and postprandial conditions in normal patients and patients with pancreatic insufficiency . Gastric juice was adjusted to pH 2, 3, 4, or 6, duodenal juice to pH 4 or 6, and bile acid concentrations to 4 or 12 mmol/L . Nutrients were added to one half of the samples . Initial bacterial or porcine lipolytic concentrations were 25 or 250 U/mL . After a 1-hour incubation at 37 degrees C, lipolytic activity was remeasured . RESULTS: In gastric juice, more bacterial lipolytic activity survived than porcine lipolytic activity at both concentrations in the absence of nutrients and at a concentration of 25 U/mL with nutrients (P < 0.05) . In duodenal juice, more bacterial lipolytic activity survived than porcine activity at pH 4 under all test conditions (P < 0.05) . CONCLUSIONS: Bacterial lipolytic activity should survive better within the gastrointestinal lumen than porcine lipolytic activity and be more effective to treat steatorrhea. Hum Mol Genet, 1994 Jul, 3(7), 1103 - 8 Identical genotypes in siblings with different homocystinuric phenotypes: identification of three mutations in cystathionine beta-synthase using an improved bacterial expression system; de Franchis R et al.; We determined the molecular basis of cystathionine beta-synthase (CBS) deficiency in three siblings with pyridoxine responsive homocystinuria using a significantly improved mutation screening method in bacteria . The phenotypic expression of the siblings differed even though their CBS genotypes were identical . The paternal allele contained a linked pair of mutations, C233G and G306C, corresponding to P78R and K102N in the polypeptide chain . Together, these inactivated the enzyme; however, expressed separately, they reduced activity by about one half . The single maternal mutation G715A (E239K) effectively abolished CBS activity . Subunits of CBS were absent from patient fibroblast extracts; however, E . coli, transformed with plasmids containing patient CBS cDNA, expressed the subunits, although in reduced amounts . The mother, an obligate heterozygote, was free from all signs of homocystinuria; nonetheless, extracts of her fibroblasts were devoid of CBS protein and activity . We conclude that fibroblast levels of CBS are only partially effective as prognosticators of disease severity and that it is important to test the in vivo response to vitamin B6 in all cases of homocystinuria, including those in which the mutations lead to the absence of the enzyme in cultured fibroblasts. Cell Mol Biol (Noisy-le-grand), 1994 Jul, 40(5), 635 - 44 Overproduction of bacterial chaperones improves the solubility of recombinant protein tyrosine kinases in Escherichia coli; Caspers P et al.; The production and purification of recombinant proteins from E . coli expression systems is often complicated by the fact that a significant amount of the foreign protein is deposited in the cytoplasm of the bacteria as aggregates or inclusion bodies . Many non-receptor protein tyrosine kinases are typical examples of proteins that are poorly soluble when overproduced in E . coli . Here, we report on the engineering of bacterial strains which overproduce chaperone proteins of the Hsp70 (DnaK, DnaJ and GrpE) and Hsp60 (GroEL and GroES) families . The simultaneous overproduction in E . coli of the chaperones DnaK, DnaJ and GrpE on the one hand, or GroEL and GroES on the other hand, and the human non-receptor protein tyrosine kinases Csk, Fyn or Lck resulted in increased solubility of the recombinant kinases . This provides the basis for future successful production and purification of large quantities of soluble and active non-receptor protein tyrosine kinases from E.coli expression systems and will enable the further characterization of these important enzyme families at the molecular level. Am J Vet Res, 1994 Jul, 55(7), 957 - 64 Comparison of species and numbers of bacteria in concurrently cultured samples of proximal small intestinal fluid and endoscopically obtained duodenal mucosa in dogs with intestinal bacterial overgrowth; Delles EK et al.; Concurrent bacterial culturing of duodenal/proximal jejunal fluid and duodenal mucosa was performed on 2 occasions in each of 16 IgA-deficient German Shepherd Dogs with small intestinal bacterial overgrowth . The interval between sample collections in each dog was 74 to 78 days . Species of bacteria and numbers of bacterial colony-forming units (CFU) per milliliter of fluid were compared with species and numbers found in the concurrent duodenal mucosa sample . There was inconsistent correlation for number of CFU and minimal correlation for species of bacteria isolated from the 2 sites . Fewer bacterial CFU usually were isolated from the mucosa than from the concurrent fluid sample . When the same numeric criteria used for diagnosing small intestinal bacterial overgrowth in samples of intestinal fluid (ie, > or = 10(5) bacterial or > or = 10(4) anaerobic CFU/ml) were used to interpret results of culturing duodenal mucosa, quantitations of bacterial CFU in duodenal mucosa was found to be a specific, but insensitive test. J Neurol Sci, 1994 Jul, 124(2), 156 - 62 Bacterial endotoxins impair electrophysiological properties of cultured astrocytes but not of cultured neurons; Koller H et al.; The endotoxins of bacteria are lipopolysaccharides which are released in the central nervous system during bacterial meningitis . Endotoxin titers in cerebrospinal fluid correspond to the appearance of severe neurological symptoms like seizures and coma . The pathogenic mechanism, however, by which endotoxins disturb neuronal function, is unclear . The functional deficit may originate either from direct alteration of neuronal excitability or from indirect effects mediated by glial cells . Therefore, we investigated the effects of lipopolysaccharides on electrophysiological properties of cortical neurons and astrocytes in separate cell cultures . Membrane potential, resistance and membrane currents of neurons were unaffected . By contrast, astrocytes depolarized markedly in a dose dependent manner (concentration range 1.0-10.0 micrograms/ml) . The depolarization was Na+ dependent and amiloride sensitive (250 microM), both indicating an activation of an electrogenic sodium dependent transport system like the Na+/Ca2+ exchanger as a source of the depolarization . These results suggest that endotoxin induced neurological deficits are not caused by direct effects on neurons, but may result from an impaired glial cell function. Radiats Biol Radioecol, 1994 Jul-Oct, 34(4-5), 578 - 81 {Effect of bacterial preparations on the survival of irradiated animals}; Mal'tsev VN et al.; A comparison of therapeutic effects of bacterial preparations (vaccinum dysenteriae, colibacterinum, bifidumbacterinum, bificolum) after gamma-irradiation was carried out . Bificolum showed the maximum and bifidumbacterinum the minimum effect, while colibacterianum had a therapeutic effect only within a narrow range of doses . Vaccinum dysenteriae was also effective. Biotech Histochem, 1994 Jul, 69(4), 199 - 202 Enhanced staining of bacterial flagella using aged mordant in the silver stain; Finegan SM et al.; Intensity of bacterial flagella staining using a modified silver stain was increased by aging the mordant for one week at room temperature . The use of aged mordant increased the apparent diameters of stained flagella and resulted in a darker stain . The mordant remained stable for at least four months at room temperature . The staining protocol presented allows application to liquid or solid cultures. Circ Shock, 1994 Jul, 43(3), 137 - 53 Human responses to bacterial endotoxin; Burrell R; The literature concerning human responses to bacterial endotoxin, in particular to the purified lipopolysaccharide derivative, has been critically reviewed and summarized . Papers selected for review are those that reported studies of human subjects in a normal state of health administered defined doses of either intravenous or aerosol inhaled material . Emphasis was placed on cardiovascular, pulmonary, metabolic, and inflammatory parameters . The information detailed here can be applied to understanding the pathophysiology of human consequences to spontaneous, clinical diseases induced by this highly inflammatory and ubiquitous substance. Appl Microbiol Biotechnol, 1994 Jul, 41(5), 574 - 7 Bacterial toxicity of cyclodextrins: luminuous Escherichia coli as a model; Bar R et al.; The effect of various concentrations of natural and chemically modified cyclodextrins on the luminescence of an Escherichia coli suspension was investigated . All cyclodextrins were found to reduce, albeit to a varying degree, the luminescence level of the bacterial cells, thus suggesting a direct interaction between the cyclodextrins and cells . The inhibitory concentrations IC20 and IC50 of the various cyclodextrins were determined and taken to represent their toxicity effects upon the bacterial cells . Among the natural cyclodextrins, gamma- and alpha-CD interfered minimally with the bacterial luminescence and consequently were essentially non-toxic . The following descending order of toxicity was observed: beta-CD >> alpha-CD > gamma-CD . Among the chemically modified cyclodextrins, Dimeb was clearly toxic while Trimeb and the hydroxylated derivatives (hydroxypropyl-alpha-CD, HPACD; -beta-CD, HPBCD; -gamma-CD, HPGCD) were essentially non-toxic . The following descending order of toxicity was observed: Dimeb >> HPBCD > Trimeb > HPACD > HPGCD. Wiad Lek, 1994 Jul, 47(13-14), 510 - 3 {Vaccine therapy in the treatment of nasal and sinus bacterial allergies}; Ziuzio S et al.; An analysis was performed of the course of nasal and sinus bacterial allergy in 160 children treated in the years 1981-1990 with bacterial vaccines . History data, results of medical examinations and laboratory investigations, indications and contraindications to vaccine therapy, and the methods and results of treatment are presented . Vaccines available in Poland were used--Polyvaccium (nasal and subcutaneous) produced by Biomed, Broncho-Vaxom, IRS-19 . As the main criterion of positive results, improvement or regression of clinical manifestations, normalization of immunoglobulin level, and also extinction of organ reactions and generalized and local skin reactions to bacterial allergens, were accepted . Very good and good results were achieved in 63% of the studied patients . The best results were achieved using combined treatment--vaccine therapy, anti-inflammatory drugs, and symptomatic antiallergic treatment (68% of positive results) . In comparison to the control group (80 children) in which no vaccine therapy was used, the results obtained in the studied group were statistically significantly better. Bioorg Khim, 1994 Jul, 20(7), 782 - 9 {(252)Cf-plasma desorption mass spectra of bacterial oligosaccharides}; El'kin IuN et al.; 252Cf plasma desorption mass spectrometry on a "reflect" desorption instrument (MSBX) was used to study oligosaccharides from bacterial O-specific antigenic chains . The data obtained for galacturonic acid, neutral and aminouronic oligosaccharides are discussed . This method is suggested for determining the composition of oligosaccharides from the O-specific polysaccharides and for elucidating their structures after modifications.
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