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