Eur J Pharmacol, 1993 Aug 3, 239(1-3), 23 - 30 Nitric oxide, but not interleukin-1, mediates the local blood flow response to lipopolysaccharide in rabbit skin; Pons F et al.; Lipopolysaccharide can mimic many aspects of the inflammatory response to gram-negative bacteria . In rabbit and rat skin, lipopolysaccharide induces neutrophil accumulation and an increase in blood flow . Interleukin-1, tumor necrosis factor-alpha and the vasodilator, nitric oxide (NO) have been implicated in the biological responses to lipopolysaccharide and gram-negative bacteria . In the present study, we characterized the local vascular response to E . coli lipopolysaccharide in rabbit skin and investigated its dependence on de novo protein synthesis, NO, interleukin-1 and neutrophils . The local vascular response to the intradermal injection of lipopolysaccharide was determined by measuring changes in local blood flow with a laser-Doppler flowmeter . Injections of lipopolysaccharide (3, 10 and 30 micrograms/site) induced a dose-related increase in blood flow, with a maximum at 2 h . The response to 10 micrograms/site lipopolysaccharide was abolished by co-injection of actinomycin D (4 x 10(-9) mol/site), reduced by 60% by NG-nitro-L-arginine methyl ester (10(-7) mol/site) and suppressed by 82% in rabbits pretreated with dexamethasone (2 mg/kg, i.v.) . However, the lipopolysaccharide-induced increase in blood flow was not affected by co-injection of recombinant human interleukin-1 receptor antagonist (7.0 x 10(-11) mol/site) or by systemic neutrophil depletion . These results demonstrate that lipopolysaccharide causes delayed vasodilatation of the rabbit cutaneous microvasculature which depends on protein synthesis and involves in part the release of NO . However, this response does not depend on interleukin-1 or neutrophils . The vasodilator effect of lipopolysaccharide may involve the expression of the inducible form of NO synthase in the vessel wall, through a mechanism independent of interleukin-1. J Laryngol Otol, 1993 Aug, 107(8), 726 - 9 Severe cochlear dysplasia causing recurrent meningitis: a surgical lesson; Stevenson DS et al.; Meningitis may be the sole presenting sign of a cerebrospinal fluid (CSF) fistula of the temporal bone . An eight-year-old boy suffering from recurrent meningitis was found to have bilateral severe cochlear dysplasia . Bilateral tympanotomies were performed, planning to obliterate each vestibule . In the right ear a stapedectomy was performed, resulting in a torrential 'CSF gusher' and difficulty in packing the vestibule . CSF rhinorrhoea requiring revision surgery and two episodes of gram-negative bacterial meningitis complicated the post-operative management, resulting in a prolonged hospital stay . Subsequently, the left ear was managed in a different fashion, leaving the stapes in situ, with grafts placed to seal the oval window niche . We would recommend this alternative procedure in cases of severe cochlear dysplasia, where abnormalities of the vestibule and basal turn of the cochlea mean that performing a stapedectomy to pack the vestibule may result in a severe 'CSF gusher', by opening directly into the subarachnoid space. Pediatr Nephrol, 1993 Aug, 7(4), 455 - 6 Hypogammaglobulinemia and fatal sepsis in an infant maintained on peritoneal dialysis; Neu AM et al.; Chronic peritoneal dialysis (CPD) is a common form of renal replacement therapy in children . Recent studies suggest that immunological abnormalities, in particular hypogammaglobulinemia, may develop in children and infants on peritoneal dialysis . We report an infant maintained on CPD who died of gram-negative sepsis . At post-mortem examination, he was noted to have severe panhypogammaglobulinemia. Am J Kidney Dis, 1993 Aug, 22(2), 296 - 9 Gentamicin clearance during hemodialysis: a comparison of high-efficiency cuprammonium rayon and conventional cellulose ester hemodialyzers; Agarwal R et al.; The advent of high-efficiency hemodialyzers has afforded improved efficiency of urea clearance; however, increased clearance of other substances, particularly antibiotics, also may occur, necessitating changes in clinical practice . Accordingly, we compared the efficiency of gentamicin removal using two different hemodialyzers, a conventional saponified cellulose ester (CD 135) and a high-efficiency cuprammonium rayon dialyzer (TAF 175L), in eight hospitalized patients undergoing antibiotic therapy for suspected or proven gram-negative infection . The rate of dialysis, estimated as the ratio of dialyzer urea clearance (K) to urea distribution volume (V) (K/V urea), and the total elimination rate constant (k) of gentamicin were measured during 17 hemodialysis treatments . The K/V urea for the two dialyzers, TAF 175L and CD 135, was 0.390 +/- 0.024 hr-1 and 0.413 +/- 0.129 hr-1 (P = NS), respectively . The TAF 175L hemodialyzer was almost twice as efficient in removing gentamicin as the CD 135: TAF 175, k = 0.263 +/- 0.024 hr-1; CD 135, k = 0.132 +/- 0.027 hr-1 (P < 0.001) . Moreover, the rate of dialysis (K/V urea) was correlated with k of gentamicin for the TAF 175L dialyzer (r2 = 0.50, P < 0.02) but not for the CD 135 dialyzer . We conclude that dialyzer characteristics and the rate of dialysis (K/V urea) should be taken into consideration when determining the dosage of gentamicin in patients on hemodialysis. Dig Dis Sci, 1993 Aug, 38(8), 1530 - 6 Alterations in rat intestinal transit by morphine promote bacterial translocation; Runkel NS et al.; Translocation of enteric microorganisms from the intestinal tract to extraintestinal sites has been proposed as an early step in the development of gram-negative sepsis . This study examined the role of altered bowel transit in influencing intestinal bacteriostasis and bacterial translocation using morphine as a pharmacologic inhibitor of such transit . In the first experiment, either normal saline (N = 8) or morphine sulfate (20 mg/kg; N = 8) was injected subcutaneously . Two hours later, morphine (7.5 mg/kg) was infused subcutaneously for an additional 22 hr; control animals received saline alone . After completion of this regimen, a volume of 0.2 ml of 2.5 mM FITC dextrans (10,000 daltons) were injected intraduodenally in each group . The bowel was removed 25 min later, divided into 5-cm segments, and the content of dextrans measured . Small bowel propulsion was expressed as the geometric center of the distribution of dextrans throughout the intestine (in percentage length of small bowel) . Gut propulsion was significantly reduced after morphine treatment as compared to controls (32.8 +/- 8.2% vs . 55.8 +/- 4.0%; P < 0.01) . In 16 additional rats, saline or morphine was again administered as described . After 24 hr, samples were obtained from the mesenteric lymph node (MLN) complex, blood, spleen, liver, duodenum, jejunum, ileum, and cecum for standard bacteriology . The bacterial counts increased significantly in each intestinal segment following morphine treatment . Microorganisms translocated to the MLN complex in 5, and to distant sites in four of eight morphine-treated animals, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) Surgery, 1993 Aug, 114(2), 140 - 6 Role of bactericidal permeability-increasing protein in the treatment of gram-negative pneumonia; Kelly CJ et al.; BACKGROUND . Gram-negative infections are a major cause of morbidity and death . Bactericidal permeability-increasing protein (BPI) is an endotoxin-neutralizing protein that also exhibits potent bactericidal activity . This study compared the efficacy of a 23 kd recombinant N-terminal fragment of BPI (rBPI23) with that of antiendotoxin antibody E5 in a model of gram-negative sepsis . METHODS . Sixty Swiss-Webster mice (Carworth farm) received an intratracheal inoculation of Escherichia coli (7 x 10(6) colony-forming units) and were randomized to three groups (20 per group) . Starting immediately after inoculation, the groups received either rBPI23 (4 mg/kg intravenously every 2 hours for four doses), E5 (11 mg/kg intravenously every 24 hours for two doses), or an isotype control antibody B55 (11 mg/kg intravenously every 24 hours for two doses) and were followed up for survival . In a second survival study, 40 mice received the same intratracheal inoculation of E . coli and were randomized to two groups . Starting 2 hours after inoculation, the groups received either rBPI23 (4 mg/kg intravenously every 2 hours for four doses) or E5 (8 mg/kg intravenously every 12 hours for four doses) and were followed up for survival . In a third study, mice received an intratracheal inoculation of 3 x 10(6) colony-forming units E . coli, a sublethal dose, and were killed to determine pulmonary and blood clearance of bacteria . RESULTS . rBPI23 conferred significantly greater protection from death than either E5 or B55 when started immediately (95% survival vs 20% and 10%, respectively; p < 0.001) or 2 hours after inoculation (65% survival vs 25% for E5; p < 0.05) . Both pulmonary and vascular clearance of bacteria was enhanced significantly by treatment with rBPI23 . CONCLUSIONS . rBPI23 may be a novel therapeutic agent in the management of gram-negative sepsis. Crit Care Med, 1993 Aug, 21(8), 1233 - 41 Introduction of new technology into critical care practice: a history of HA-1A human monoclonal antibody against endotoxin; Luce JM; OBJECTIVES: HA-1A, a monoclonal antibody against endotoxin, was thought to be effective in treating patients with Gram-negative sepsis . Because of this possibility, many clinicians felt obligated to use the drug and assumed that its product license application would be approved by the U.S . Food and Drug Administration (FDA) . Nevertheless, the efficacy of HA-1A was not conclusively demonstrated by a first clinical trial . The FDA rejected the product license application and requested a second clinical trial, which was suspended after excess mortality was noted in patients treated with HA-1A . This review of the history of the drug was prepared to provide clinicians and sepsis investigators with information about HA-1A and, by extension, the process by which new technology is introduced into critical care practice . DATA SOURCES: Data used to prepare this review were obtained from the author's personal files as well as the computerized MEDLINE database . STUDY SELECTION: Studies were selected for their relevance to the history of HA-1A and their relevance to the introduction of potentially useful medical technology . DATA EXTRACTION: The author extracted all applicable data . DATA SYNTHESIS: Although the first clinical trial of HA-1A suggested that the drug was effective in treating patients with Gram-negative bacteremia with or without shock, further analysis by the FDA indicated a benefit only for bacteremic patients with shock . Furthermore, the original study design was not followed, leading in part to the FDA's refusal of the product license application . Concern also was raised over the issue of identifying which patients should receive HA-1A and the cost of the drug, which would have put it past the reach of some American hospitals and thereby, would have conflicted with the ethical principle of social justice . Finally, the second trial suggested that HA-1A might be harmful . CONCLUSIONS: Due to the FDA's action, the issues raised about HA-1A, and the results of the two clinical trials, clinicians should not use the drug . The history of HA-1A provides insights about how new technology is and will be introduced into critical care practice. J Infect Dis, 1993 Aug, 168(2), 473 - 6 Fluid administration, brain edema, and cerebrospinal fluid lactate and glucose concentrations in experimental Escherichia coli meningitis; Tauber MG et al.; The effect of no fluids versus liberal fluid supplementation on brain edema and cerebrospinal fluid (CSF) lactate and glucose concentrations was compared in rabbits with experimental Escherichia coli meningitis . Fluid restriction for the duration of the experiment (19 h) led to a decrease in body weight by approximately 5%, while the high fluid regimen increased body weight by approximately 5% . Infected animals developed brain edema compared with controls, but the fluid regimen had no measurable effect on the degree of edema . In contrast, fluid-restricted animals had significantly higher CSF lactate and lower CSF glucose concentrations than fluid-supplemented animals (lactate, 13.5 +/- 3.5 vs . 10.1 +/- 3.3 mmol/L; glucose, 1.89 +/- 1.39 vs . 4.11 +/- 1.39 mmol/L) . These results fail to support the hypothesis that administration of large amounts of fluid in this model of gram-negative bacterial meningitis aggravates brain edema. J Bacteriol, 1993 Aug, 175(15), 4911 - 6 Site-directed mutations in the relaxase operon of RP4; Cole SP et al.; Mutations were constructed by site-directed mutagenesis in the relaxase operon of the broad-host-range plasmid RP4 . The mutations were constructed in smaller plasmids, recombined into the 60-kb RP4 plasmid, and tested for their ability to transfer . The relaxase operon contains the transfer genes traJ, traH, and traI, which are involved in nicking at the transfer origin to generate the single strand destined to be transferred to the recipient cell . In the first mutant, the C terminus of TraI was truncated, leaving TraH intact . This mutant decreased transfer by approximately 500-fold in Escherichia coli, and the traI mutation could be complemented by a wild-type copy of traI in trans in the donor . The traI mutation similarly decreased transfer between a variety of gram-negative bacteria . A site-specific mutation was made by the polymerase chain reaction-based unique-site mutagenesis procedure to alter the start site of traH . This mutation had no effect on intraspecific E . coli transfer but reduced transfer by up to sevenfold for some gram-negative bacteria . The traH mutation had no effect on plasmid stability . Thus, neither TraH nor the C terminus of TraI is required for conjugative transfer, but both increase mating efficiency in some hosts. J Bacteriol, 1993 Aug, 175(15), 4756 - 63 Oar, a 115-kilodalton membrane protein required for development of Myxococcus xanthus; Martinez-Canamero M et al.; Myxococcus xanthus is a developmental gram-negative bacterium which forms multicellular fruiting bodies upon nutrient starvation . This bacterium was found to contain a 115-kDa membrane protein which separated with the inner membrane fraction by sucrose density gradient centrifugation . The gene for this protein was cloned, and its DNA sequence was determined . The deduced amino acid sequence consists of 1,061 residues . This protein contains a putative signal sequence and many short segments, found scattered throughout the entire protein, that have sequence similarities with OmpA, a major outer membrane protein of Escherichia coli . Thus, the gene was designated oar (OmpA-related protein) . A second open reading frame was found 36 bases downstream of the oar termination codon . This open reading frame encodes a protein of 236 residues and contains a putative lipoprotein signal sequence . An aor disruption mutation (delta oar) showed no effect on vegetative growth but caused abnormal morphogenesis during development and reduced myxospore formation . When examined with a light microscope, delta oar cells were unable to aggregate on developmental agar, indicating that Oar is required for cellular adhesiveness during development. Infect Immun, 1993 Aug, 61(8), 3184 - 9 An interleukin-6-induced acute-phase response does not confer protection against lipopolysaccharide lethality; Bucklin SE et al.; Lipopolysaccharide (LPS), a component of gram-negative bacterial outer cell walls, can stimulate lymphoreticular cells to produce cytokines such as tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1), and IL-6 . One of these proinflammatory cytokines, IL-6, induces hepatic synthesis of a class of proteins termed acute-phase proteins . D-Galactosamine inhibits acute-phase protein synthesis and concurrently sensitizes mice to a lethal dose of LPS approximately 10,000-fold . From these observations, we hypothesized that the acute-phase response may serve as a defense mechanism for protection of the host against the deleterious effects of LPS . To test this hypothesis, murine recombinant IL-6 (mrIL-6) was used to induce an acute-phase response prior to a lethal LPS challenge in both D-galactosamine-treated and normal mice . Induction of the acute-phase response by mrIL-6 was quantitated by measuring the concentrations of fibrinogen and complement component C3, two well-characterized acute-phase proteins, in the circulation . The effect of acute-phase and normal serum on TNF-alpha release by peritoneal macrophages stimulated with LPS in vitro was also examined . The results of these studies confirmed the induction of the acute-phase response by mrIL-6, as reflected in an approximate doubling in circulating levels of fibrinogen and C3 . However, when either D-galactosamine-sensitized or normal mice were challenged with a lethal dose of LPS at various times after mrIL-6 administration, the acute-phase response induced by mrIL-6 did not alter either cumulative lethality or the kinetics of lethality . Additionally, compared with normal serum, acute-phase serum did not affect TNF-alpha release by peritoneal macrophages following LPS-mediated stimulation in vitro . Collectively, these studies would not support a dominant role for an IL-6-mediated acute-phase response as contributing to the resistance of normal mice compared with D-galactosamine-sensitized mice in LPS-induced lethal toxicity. Eur J Med, 1993 Aug-Sep, 2(7), 404 - 7 Acid-base and electrolyte abnormalities in febrile patients with bacteraemia; Elisaf M et al.; OBJECTIVES: Very commonly febrile patients with bacteraemia develop a variety of acid-base and electrolyte disturbances which play a significant role in the morbidity and mortality of these patients . This study was undertaken to describe the pathogenetic mechanisms of these abnormalities in febrile patients with bacteraemia . METHODS: Fifteen febrile patients with bacteraemia, aged 24-62 years, were studied . In all patients blood cultures revealed Gram-negative rods . None of them had septic shock, diabetes mellitus, renal or liver failure and none was receiving drugs influencing acid-base balance and electrolyte levels or was a heavy alcohol consumer . RESULTS: Nine patients had respiratory alkalosis, which was possibly due to bacterial toxins, while the remaining 6 had a wide-gap metabolic (lactic) acidosis coexisting with respiratory alkalosis . Hypokalaemia was found in four patients and was mainly due to respiratory alkalosis . However, kaliuria due to hypomagnesaemia contributed to hypokalaemia in 2 patients . Hypomagnesaemia was detected in 3 patients and was attributed to respiratory alkalosis as well as to magnesiuria induced by metabolic acidosis or phosphate depletion . Hypophosphataemia was found in 5 patients who also had respiratory alkalosis and/or phosphaturia due to metabolic acidosis or hypomagnesaemia . Finally, one patient had multifactorial origin hypocalcaemia . CONCLUSION: Febrile patients with bacteraemia develop a number of acid-base and electrolyte disturbances attributed to various pathogenetic mechanisms. Diagn Microbiol Infect Dis, 1993 Aug-Sep, 17(2), 163 - 6 Central venous catheter infection caused by Moraxella osloensis in a patient receiving home parenteral nutrition; Buchman AL et al.; We report the first case of a central venous catheter infection caused by Moraxella osloensis, which was successfully treated without catheter removal . The isolation, identification, and pathogenesis of this species are discussed . It is recommended that Moraxella isolates be identified to species in order to determine the relative pathogenic and opportunistic roles of the various Moraxella species . Our case also demonstrates that catheter sepsis caused by some Gram-negative organisms may be amenable to systemic antibiotic therapy without the necessity of catheter removal. Thorax, 1993 Aug, 48(8), 785 - 9 Factors of importance for the long term prognosis after hospital treated pneumonia; Hedlund JU et al.; BACKGROUND--Elderly patients admitted to hospital for community acquired pneumonia have a high risk of recurrence of pneumonia and of death during the years after discharge . In this study potential factors of importance for the long term prognosis after hospital treated pneumonia were retrospectively investigated . METHODS--A total of 241 patients (103 men) with a mean age of 60 (range 18-102) years discharged from hospital after treatment for community acquired pneumonia were studied . After an average follow up period of 31 months, 18 independent variables present during hospital treatment of the initial pneumonia were examined for association with the following end points: recurrence of pneumonia, death from any cause, and death from pneumonia . RESULTS--Age adjusted analysis showed that systemic treatment with corticosteroids correlated significantly with recurrence of pneumonia and with death . The presence of low serum albumin levels on admission or colonisation of the respiratory tract with Gram negative enteric bacteria seemed to be important negative prognostic factors for the outcome during pneumonia recurrences after discharge . CONCLUSIONS--Patients who are admitted to hospital with pneumonia are at risk of subsequent pneumonia and death after discharge . This risk seems to be even higher in patients who are treated with corticosteroids systemically, who have a low serum albumin level on admission, or who become colonised in the respiratory tract with Gram negative enteric bacteria during their hospital stay. J Rheumatol, 1993 Aug, 20(8), 1388 - 96 Responses to gram negative enteric bacterial antigens by synovial T cells from patients with juvenile chronic arthritis: recognition of heat shock protein HSP60; Life P et al.; OBJECTIVE . To investigate the antigenic specificity of synovial T cells in juvenile chronic arthritis (JCA) . METHODS . Synovial fluid and peripheral blood mononuclear cells from 24 patients with JCA were tested for their proliferative responses to recall antigens, enteric organisms associated with reactive arthritis, influenza A and a recombinant preparation of a mycobacterial heat shock protein, HSP65 . To investigate further recognition of this last antigen, synovial T cells from one B27+ patient with pauciarticular disease were cloned using HSP65 . The specificity of the resultant clones was then examined . RESULTS . Marked synovial T cell responses to enteric organisms and to HSP65 were noted, particularly in HLA-B27+, pauciarticular patients; these were similar to those seen in a B27+ patient with reactive arthritis . Responses to enteric organisms and to HSP65 were significantly correlated, suggesting recognition of an epitope common to these antigen preparations . However, all of the T cell clones obtained using HSP65 proved to recognize E . coli derived antigens contaminating the recombinant HSP65 rather than the mycobacterial antigen; these contaminants included the 60 kDa E . coli HSP, GroEL . The GroEL specific T cells did not respond to heat shocked human cells; this suggests (but does not prove) that they do not crossreact with human HSP60 . CONCLUSION . Synovial T cell recognition of antigens from enteric organisms associated with reactive arthritis is a common feature in pauciarticular JCA . Among the target antigens is the GroEL HSP, but T cells recognizing this antigen do not necessarily crossreact with the homologous human HSP60. Infect Immun, 1993 Aug, 61(8), 3149 - 56 Endotoxin-mediated endothelial cell injury and activation: role of soluble CD14; Arditi M et al.; Vascular endothelial cell (EC) injury by lipopolysaccharides (LPS) plays a major role in the pathogenesis of gram-negative bacterial sepsis and endotoxic shock . The studies described here were performed to define further the molecular mechanisms involved in the EC responses to LPS . We showed that serum was required for LPS-mediated cytotoxicity for bovine brain microvessel, pulmonary, and aortic ECs and that anti-human CD14 antibodies completely blocked LPS-mediated cytotoxicity for ECs in the presence of human serum . The addition of a recombinant soluble form of human CD14 to serum-free medium restored the LPS-mediated cytotoxicity, whereas the addition of LPS binding protein (LBP), a serum protein that potentiates LPS-induced responses to monocytes, had no effect . A similar dependency on serum or recombinant soluble CD14 (under serum-free conditions) was observed for LPS-induced secretion of interleukin-6 by human umbilical vein ECs . These findings indicate that soluble CD14 is required for LPS-mediated EC responses independently of LPB, suggesting that serum soluble CD14 represents a naturally occurring agonist for EC responses to LPS. J Immunol, 1993 Jul 15, 151(2), 916 - 21 IFN-gamma involvement in the severity of gram-negative infections in mice; Kohler J et al.; The role of IFN-gamma in the regulation of inflammation leading to gram-negative septic shock is still poorly understood . IFN-gamma blockade has been shown to improve the survival of animals challenged with i.v . bolus injections of LPS and gram-negative bacteria . We have investigated a model of focal Escherichia coli infection leading to peritonitis and septic shock . Mice were challenged i.p . with an inoculum near the LD50 . The addition of rIFN-gamma together with bacteria increased the mortality and the level of blood TNF-alpha and IL-6 . Conversely blockade of IFN-gamma with a neutralizing mAb significantly improved the survival of the mice . This beneficial effect was not associated with a stringent decrease in blood bacterial counts and TNF-alpha and IL-6 levels in survivors . In this model, the protective effect of anti-IFN-gamma mAb contrasted with the ineffectiveness of a neutralizing anti-TNF-alpha mAb . These findings suggest that overproduction of IFN-gamma might have a more detrimental role than overproduction of TNF-alpha during focal gram-negative infections. Ann Emerg Med, 1993 Jul, 22(7), 1164 - 8 Effect of needle changing and intravenous cannula collection on blood culture contamination rates; Smart D et al.; STUDY OBJECTIVES: We tested the hypotheses that blood culture positivity and contamination rates were not increased by not changing needles between venipuncture and inoculation of blood culture bottles or by taking blood for culture by freshly inserted IV cannulae . DESIGN: A prospective study of blood cultures collected by venipuncture or IV cannulae taken from an emergency department population . Venipuncture samples were randomized into needle change (standard method) or no needle change before inoculation into blood culture bottles . PARTICIPANTS: Nine hundred forty patients requiring blood cultures after assessment in the ED . INTERVENTIONS: A standard disinfection procedure using 0.5% chlorhexidine in 70% alcohol was used . Blood was collected by venipuncture and inoculated with or without needle change . Blood collected by IV cannula was inoculated with a fresh needle applied to the collection syringe . MEASUREMENTS AND MAIN RESULTS: There was no statistically significant difference in contamination rates for blood collected by venipuncture with no needle change (6.4%) compared with needle change (4.2%, P > .30) . No significant difference in contamination rates was noted for blood taken by freshly inserted IV cannulae (4.3%) compared with venipuncture with needle change after sampling (4.2%, P > .90) . Some problems with randomization resulted in unequal numbers in the needle-change (286) versus no-needle-change (141) subgroups, and this may have introduced bias . A higher rate of pathogen growth was observed in blood taken by IV cannula (11.4%) compared with the standard method (6.3%) (P < .025) . A significantly greater rate of Gram-negative sepsis was noted in the IV cannula group (6.6%) compared with direct venipuncture with needle change (1.1%) and no needle change (4.2%, P < .01) . CONCLUSION: The results of this study do not support the practice of changing needles before inoculating blood samples into blood culture bottles . Collection of blood for culture through freshly inserted IV cannulae is associated with a low contamination rate and is an acceptable alternative to direct venipuncture . Sources of bias in this study suggest that further research is needed to determine the optimal technique for collecting blood cultures. J Gen Microbiol, 1993 Jul, 139 ( Pt 7), 1531 - 5 Phylogeny and phenotypic characterization of the stalk-forming and iron-oxidizing bacterium Gallionella ferruginea; Hallbeck L et al.; The 16S rRNA gene of Gallionella ferruginea was amplified by polymerase chain reaction and sequenced by direct double-stranded sequencing . The phylogenetic analysis placed G . ferruginea in the beta-group of the Proteobacteria, with 90.0% similarity to Nitrosolobus multiformis and 88.6% to Rhodocyclus purpureus . The published phenotypic characteristics of G . ferruginea were compiled and supplemented with growth experiments using ferrous iron, thiosulphate and sulphide as electron donor, and nitrate as nitrogen source . G . ferruginea is a Gram-negative, curved bacterium with one polar flagellum . It grows auto- and mixotrophically with CO2, glucose, fructose and sucrose as carbon sources, ferrous iron as an electron donor and ammonium or nitrate as nitrogen sources . Two G . ferruginea specific oligonucleotide probes are suggested . An iron-oxidizing bacterium without stalk-forming ability, but with the same growth pattern as G . ferruginea, was identified as G . ferruginea by comparison of highly variable parts of the 16S rRNA gene . This indicates that the stalk is not essential for growth. Curr Opin Oncol, 1993 Jul, 5(4), 625 - 32 Febrile neutropenia; Klastersky J; Severe neutropenia and its related infectious complications remain a permanent threat for patients receiving intensive chemotherapy, especially in the context of bone marrow transplantation . Chemoprophylaxis and use of colony-stimulating factors have altered the severity of the clinical picture in a favorable direction: neutropenia can be shortened, and gram-negative infection can be made less frequent; neither can be yet abolished . Early therapy, eg, empiric combination treatment, remains the cornerstone of our approach to febrile neutropenia; the actual choice of agents is probably less important and should be guided by local epidemiologic conditions . The concepts of empiric therapy also starts to be more widely accepted for the control of fungal and viral infections . Finally, it is fair to recognize that, at the other end of the spectrum of febrile neutropenia, conventional chemotherapy that results in only moderate and short neutropenia can usually be managed without much problem, namely with broad-spectrum monotherapy . Other possible simplified approaches should be investigated under controlled conditions and in patients selected on the basis of favorable prognostic factors. Pharmacotherapy, 1993 Jul-Aug, 13(4), 330 - 9 The role of Helicobacter pylori in peptic ulcer disease; Partipilo ML et al.; The pathophysiology of peptic ulcer disease (PUD) is often described as an imbalance between aggressive factors such as acid and pepsin and alterations in the mucosal protective mechanisms . Helicobacter pylori is a gram-negative organism that has been identified as a potential causative agent in the pathogenesis of PUD . The exact mechanism by which it contributes to mucosal damage is unknown . It is thought that the organism may disrupt the protective mucous layer, allowing the underlying epithelium to be injured by gastric acid . Significant evidence indicates that H . pylori is a major etiologic factor in type B gastritis . Data confirming its etiologic role in duodenal ulcer (DU) disease is not conclusive; however, eradication of the organism is associated with a reduction in the recurrence of DU . Optimum therapy to eradicate H . pylori has not been established, although several multidrug regimens have been evaluated . Treatment of H . pylori infection should be reserved for individuals in whom conventional therapy for DU is unsuccessful and those whose ulcers relapse during maintenance therapy. Appl Environ Microbiol, 1993 Jul, 59(7), 2077 - 81 Isolation and characterization of Selenomonas ruminantium strains capable of 2-deoxyribose utilization; Rasmussen MA; Microbes from ruminal contents of cattle were selectively enriched by using 2-deoxyribose (2DR) as a substrate for growth . Bacterial isolates growing on 2DR were gram-negative, curved, motile rods . The isolates grew on a broad range of substrates, including deoxyribose, glucose, ribose, mannitol, and lactate as well as ribonucleosides and deoxyribonucleosides . The strains also grew on rhamnose (6-deoxymannose) but not DNA . Organic acids produced from growth on hexoses and pentoses included acetate, propionate, lactate, and succinate . The isolates were identified as Selenomonas ruminantium subsp . lactilytica on the basis of morphology, substrate specificity, and other biochemical characteristics . Several characterized species of ruminal bacteria were also screened for growth on 2DR, with only one strain (S . ruminantium PC-18) found able to grow on 2DR . Ethanol was produced by 2DR when strains were grown on ribose or 2DR. Radiographics, 1993 Jul, 13(4), 787 - 96 Imaging of complications of lung transplantation; O'Donovan PB; With the increasing number and improved survival of lung transplant recipients, radiologists should be aware of the imaging features of lung transplants and the associated complications . Reimplantation response, a noncardiogenic pulmonary edema seen 48 hours after transplantation that subsequently resolves, varies in appearance from a mild perihilar haze to a dense consolidation in the perihilar areas and lung bases . A late complication of omentopexy (used to prevent bronchial dehiscence) is herniation of abdominal contents through the diaphragmatic incision into the thorax . Extrabronchial air collections are a radiologic manifestation of anastomotic dehiscence . Stricture formation that compromises the bronchial lumina is usually visible with plain radiography, but computed tomography can aid in the evaluation . Acute rejection is evident radiographically as new or increasing pleural effusions, septal lines, subpleural edema, peribronchial cuffing, and air-space disease, without increase in cardiac size . Radiographic features of chronic rejection include both increased and diminished lung volumes, central and peripheral bronchiectasis, localized air-space disease, partial lobar atelectasis, thin linear irregular areas of increased opacity, pleural thickening, and diminished peripheral lung markings . Infection is frequently seen, especially gram-negative pneumonias, with fewer occurrences of cytomegalovirus infection, candidiasis, and invasive aspergillosis. Parasitology, 1993 Jul, 107 ( Pt 1), 49 - 53 Endotoxins and the pathogenesis of Trypanosoma brucei brucei infection in mice; Alafiatayo RA et al.; The involvement of endotoxins in Trypanosoma brucei brucei infection in CD-1 mice was investigated by the Limulus amoebocyte lysate (LAL) test . At 7 days post-infection mean serum endotoxin level was elevated by 2.5 times (36.4 pg/ml cf . control 14.25 pg/ml, P < 0.001) and a similar increase was maintained throughout the infection (survival 28-35 days) . Purified disrupted parasites contained significant endotoxin activity (mean value 280 pg/mg protein) . The mouse infections were also associated with progressive Gram-negative bacteraemia (present in 4 out of 5 infected animals by day 28 p.i.) . The increased endotoxin levels may be due to parasite products, the products of intercurrent bacterial infections, other unidentified sources (e.g . from the gut), or a combination of these . It is concluded that the raised endotoxins may be important contributive factors in the pathogenesis of experimental murine trypanosomiasis. Infect Control Hosp Epidemiol, 1993 Jul, 14(7), 383 - 9 A prospective randomized trial comparing manual and automated endoscope disinfection methods; Fraser VJ et al.; OBJECTIVE: To compare the efficacy of endoscope disinfection using automated and manual systems . DESIGN: Prospective randomized trial . SETTING: A 1,000-bed tertiary care referral center . METHODS: All endoscopes underwent a three-stage decontamination process including brushing and cleaning with water and detergent, manual or automated disinfection with 2% glutaraldehyde, and 70% alcohol rinse with forced air drying . Cultures were obtained from endoscopes from both groups before and after alcohol rinse and then after overnight storage . RESULTS: Cultures from 8/30 (27%) automated and 11/30 (37%) manually disinfected (P = 0.58) endoscopes grew gram-negative bacteria and/or nontuberculous mycobacteria before the alcohol rinse . After alcohol rinse, 3 (10%) of 30 automated and 8 (27%) of 30 manually disinfected endoscopes remained contaminated (P = 0.28) . Manually disinfected endoscopes were contaminated more frequently with coliform bacteria, whereas endoscopes undergoing automated disinfection were more frequently contaminated with nontuberculous mycobacteria, but the differences were not statistically significant . After alcohol rinse and forced air drying, there was no difference in contamination rates between freshly disinfected endoscopes and those stored overnight (7/30 (23%) versus 4/30 (13%), P = 0.50) . Colonoscopes and duodenoscopes were contaminated more often than gastroscopes (P = 0.00001) . CONCLUSION: The persistent endoscope contamination after manual and automated disinfection indicates the importance of developing more reliable and effective disinfection methods. J Clin Microbiol, 1993 Jul, 31(7), 1936 - 9 Use of immunoelectron microscopy to demonstrate Francisella tularensis; Geisbert TW et al.; Three immunoelectron microscopy (IEM) methods were employed to show laboratory-cultivated Francisella tularensis . By the IEM assays, F . tularensis was distinguished from four antigenically distinct gram-negative bacteria . IEM should be a valuable tool for confirming presumptive isolates of F . tularensis and may potentially be useful for demonstrating other medically important bacteria. J Leukoc Biol, 1993 Jul, 54(1), 81 - 8 Lipopolysaccharide-induced suppression of erythrocyte binding and phagocytosis via Fc gamma RI, Fc gamma RII, Fc gamma RIII, and CR3 receptors in murine macrophages; Sundaram R et al.; In this study we have investigated the ability of lipopolysaccharide (LPS) to suppress binding and phagocytosis of erythrocytes via various receptors on mouse macrophages . Thioglycollate-elicited peritoneal macrophages were treated in vitro with LPS and the ability to bind and phagocytose radiolabeled sheep red blood cells was determined . We show that LPS can directly suppress phagocytosis of immunoglobulin G-opsonized and nonopsonized sheep red blood cells (SRBCs) by inflammatory macrophages . Suppression was dose dependent and was observed after 4 h of exposure . This effect lasted for at least 24 h following the removal of LPS . LPS suppressed the binding, rate, and absolute level of phagocytosis via Fc receptors . Phagocytosis via all Fc receptors (Fc gamma RI, Fc gamma RII, and Fc gamma RIII) was suppressed by LPS . Furthermore, suppression was not limited to Fc receptor-mediated phagocytosis because binding and uptake of C3bi-opsonized SRBCs to CR3 receptors was also decreased following LPS treatment . LPS did not exert its effects via the production of interleukin-1 (IL-1), IL-6, tumor necrosis factor alpha, or interferon alpha/beta, because antibodies to these cytokines did not abrogate the effect . The ability of LPS to suppress binding and phagocytosis of microorganisms may contribute to the toxic effects of LPS during gram-negative sepsis by preventing or delaying elimination of bacteria by host macrophages. Ann Surg, 1993 Jul, 218(1), 79 - 90 Comparison of peripheral blood leukocyte kinetics after live Escherichia coli, endotoxin, or interleukin-1 alpha administration . Studies using a novel interleukin-1 receptor antagonist; Hawes AS et al.; OBJECTIVE: This study was undertaken to evaluate whether hematologic and immunologic effects observed after bacteremia and endotoxemia in the host could be replicated by administration of recombinant human interleukin-1 alpha (IL-1 alpha) in a primate model . Furthermore, to determine whether endogenously produced interleukin-1 (IL-1) contributes to the changes observed during endotoxemia or gram-negative septic shock, a specific IL-1 receptor antagonist (IL-1 ra) was administered . SUMMARY BACKGROUND DATA: The lipopolysaccharide (LPS) component of the outer membrane of gram-negative bacteria initiates a constellation of metabolic and immunologic host responses . IL-1, a macrophage-derived cytokine, acts as a key mediator in the host response to infection and inflammation . METHODS: Baboons were randomly assigned to receive either recombinant human IL-1 alpha, LPS, or live Escherichia coli both with or without concomitant administration of IL-1ra . Blood was collected hourly and analyzed using flow cytometric techniques . RESULTS: Both endotoxemia and live E . coli bacteremia induced an acute granulocytopenia; however, the granulocytopenia gradually resolved in the endotoxemic group, but was sustained in the bacteremic group . An early lymphopenia and monocytopenia was elicited by LPS or E . coli and persisted throughout the experiment . Recombinant human IL-1 alpha induced the following: (1) an early, transient decline in granulocytes followed by a sustained granulocytosis; (2) a lymphopenia; and (3) a transient monocytopenia followed by a gradual return to baseline . Although IL-1ra had no effect on leukocyte kinetics with either live E . coli or LPS, the IL-1ra significantly abrogated the monocytopenia seen with recombinant human IL-1 alpha administration alone . CONCLUSIONS: These results suggest that administration of recombinant human IL-1 alpha can replicate some of the characteristic patterns of hematologic change associated with bacteremia and endotoxemia . However, an endogenous IL-1 response is not required for these changes to occur . Rather, the data suggest that other inflammatory mediators induced by endotoxemia or gram-negative bacteremia, such as tumor necrosis factor-alpha (TNF alpha), may be involved. Hosp Pract (Off Ed), 1993 Jul, 28 Suppl 2, 36 - 9; discussion 60-1 Outpatient parenteral antibiotic therapy . Management of serious infections . Part II: Amenable infections and models for delivery . Osteomyelitis; Tice AD; Osteomyelitis is one of the most common and well-established indications for outpatient parenteral antibiotic therapy . Because patients are usually otherwise healthy and therapy is prolonged (four to six weeks), this infection is especially suited to outpatient management . While most gram-negative infections in adults can be treated with an oral quinolone, others usually require IV therapy. Hepatology, 1993 Jul, 18(1), 173 - 8 Gram-negative bacterial lipopolysaccharide impairs hyaluronan clearance in vivo and its uptake by the isolated, perfused rat liver; Deaciuc IV et al.; The purpose of this investigation was to examine the effect of gram-negative bacterial lipopolysaccharide on hyaluronan concentration in blood plasma, hyaluronan removal from the blood and hyaluronan uptake by isolated, perfused rat liver . Intravenous administration of Escherichia coli lipopolysaccharide to rats markedly increased plasma hyaluronan concentration in a dose-dependent manner . One day after lipopolysaccharide challenge (0.1 or 1.0 mg per 100 gm body wt), plasma hyaluronan levels were 570.7 +/- 66.8 ng x ml-1 and 1,951.0 +/- 120.3 ng x ml-1, respectively, as compared with 94.2 +/- 12.2 ng x ml-1 in the time-matched control animals . Removal of intravenously injected hyaluronan (30 micrograms per 100 gm body wt) was suppressed 32% by lipopolysaccharide administration (100 micrograms per 100 gm body wt) . At the same dose, lipopolysaccharide induced a severe inhibition (60% to 80%) of hyaluronan uptake by perfused livers isolated 3 or 24 hr after lipopolysaccharide administration . The inhibitory effect of lipopolysaccharide on hyaluronan uptake by the isolated, perfused liver was not abolished by pretreatment with either antibodies to tumor necrosis factor-alpha IgG or indomethacin, an inhibitor of the cyclooxygenase pathway . Continuous intravenous infusion of recombinant murine tumor necrosis factor-alpha for 18 to 20 hr did not affect plasma hyaluronan concentration . These data suggest that neither tumor necrosis factor-alpha, an early cytokine induced by lipopolysaccharide, nor prostaglandins are involved in the mechanism of lipopolysaccharide-induced inhibition of hyaluronan uptake by the perfused rat liver.(ABSTRACT TRUNCATED AT 250 WORDS) J Bacteriol, 1993 Jul, 175(13), 4225 - 34 Molecular cloning and sequence analysis of the gene encoding OmpL1, a transmembrane outer membrane protein of pathogenic Leptospira spp; Haake DA et al.; Pathogenic Leptospira spp . are spirochetes that have a low transmembrane outer membrane protein content relative to that of enteric gram-negative bacteria . In a previous study we identified a 31-kDa surface protein that was present in strains of Leptospira alstoni in amounts which correlated with the outer membrane particle density observed by freeze fracture electron microscopy (D . A . Haake, E . M . Walker, D . R . Blanco, C . A . Bolin, J . N . Miller, and M . A . Lovett, Infect . Immun . 59:1131-1140, 1991) . The N-terminal amino acid sequence was used to design a pair of oligonucleotides which were utilized to screen a lambda ZAP II library containing EcoRI fragments of L . alstoni DNA . A 2.5-kb DNA fragment which contained the entire structural ompL1 gene was identified . The structural gene deduced from the sequence of this DNA fragment would encode a 320-amino-acid polypeptide with a 24-amino-acid leader peptide and a leader peptidase I cleavage site . Processing of OmpL1 results in a mature protein with a predicted molecular mass of 31,113 Da . Secondary-structure prediction identified repeated stretches of amphipathic beta-sheets typical of outer membrane protein membrane-spanning sequences . A topological model of OmpL1 containing 10 transmembrane segments is suggested . A recombinant OmpL1 fusion protein was expressed in Escherichia coli in order to immunize rabbits with the purified protein . Upon Triton X-114 extraction of L . alstoni and phase separation, anti-OmpL1 antiserum recognized a single band on immunoblots of the hydrophobic detergent fraction which was not present in the hydrophilic aqueous fraction . Immunoelectron microscopy with anti-OmpL1 antiserum demonstrates binding to the surface of intact L . alstoni . DNA hybridization studies indicate that the ompL1 gene is present in a single copy in all pathogenic Leptospira species that have been tested and is absent in nonpathogenic Leptospira species . OmpL1 may be the first spirochetal transmembrane outer membrane protein for which the structural gene has been cloned and sequenced. Am J Pathol, 1993 Jul, 143(1), 76 - 84 Murine tissue factor gene expression in vivo . Tissue and cell specificity and regulation by lipopolysaccharide; Mackman N et al.; Regulation of tissue factor (TF) gene expression was studied in vivo employing a murine model system . In untreated mice, TF mRNA was detected in brain, lung, kidney, and heart by Northern blot analysis . After administration of lipopolysaccharide, steady-state levels of TF mRNA were unchanged in brain, decreased in heart, and increased in both kidney and lung . In the brain, Bergmann glia within the Purkinje cell layer of the cerebellum and neuroglia within the cerebral cortex expressed TF mRNA by in situ hybridization . Epidermal cells of the skin and tongue also expressed TF mRNA . At present, we have not identified the cell type(s) in the kidney and lung responsible for increased TF gene expression . These results demonstrate tissue- and cell-specific TF gene expression in vivo . Lipopolysaccharide-mediated increases in TF expression in the kidney and lung may promote fibrin deposition in these organs during Gram-negative sepsis. West Afr J Med, 1993 Jul-Sep, 12(3), 180 - 4 Acute encephalopathy, hypertension and gram negative sepsis in sickle cell disease; Akpede GO et al.; The case histories of two patients with sickle cell disease and gram negative sepsis complicated by encephalopathy and hypertension is presented . The first patient had 2 episodes of "hypertensive encephalopathy" before control of her blood pressure was achieved while the second patient had only one . The occurrence, though apparently rare, can have serious implications . Possible mechanisms are discussed and the need to monitor the blood pressure of children with sickle cell disease is stressed. ASAIO J, 1993 Jul-Sep, 39(3), M773 - 7 Production of platelet activating factor by human neutrophils after backfiltration of endotoxin contaminated dialysate; David S et al.; Lipopolysaccharide (LPS) from gram negative bacteria has been shown to prime human polymorphonuclear neutrophil (PMN) production of platelet activating factor (PAF) . PAF is a lipid mediator of inflammation and endotoxic shock and is also involved in leukocyte activation occurring during hemodialysis; PAF induces leukopenia, degranulation of lysosomal granules, and adherence to hemodialysis membranes . Transmembrane passage of LPS has also been shown to occur . To evaluate the relevance of transmembrane passage of LPS on the priming of PMN derived production of PAF, we designed in vitro studies using an experimental circuit equipped with different membranes (Cuprophan, polysulfone, polymethylmethacrylate, polyamide) and recirculation of purified human PMNs . At different time intervals, PMNs were stimulated with FMLP (10 microM) after back-filtration of sterile and LPS contaminated dialysate . The results of these studies suggest that priming of PMN derived production of PAF was related to the percent of backfiltered LPS, and they emphasize the need for careful assessment of microbiologic quality to improve biocompatibility. Cytokine, 1993 Jul, 5(4), 348 - 53 Antilipid a monoclonal antibody HA-1A: immune complex clearance of endotoxin reduces TNF-alpha, IL-1 beta and IL-6 production; Katsikis P et al.; HA-1A is a human monoclonal IgM antibody which recognizes the lipid A component of lipopolysaccharide (LPS) . This antibody has reduced mortality in the septic shock syndrome resulting from Gram negative bacteria, in which many of the manifestations are considered to be due to cellular activation and secretion of cytokines, most notably TNF-alpha . However HA-1A does not directly neutralize LPS effectively in vitro, and studies reported to date have not defined its mechanism of action . Here we demonstrate that HA-1A, which in the presence of complement promotes immune adherence, may inhibit LPS action by facilitating its sequestration on red blood cells and clearance to an extent that cytokine production is reduced . Incubation of LPS at clinically significant (pg/ml) does with HA-1A at therapeutic levels (e.g . 10 micrograms/ml) and complement resulted in LPS association with erythrocyte CR1 receptors . This reduced the ability of the residual, free LPS by 50-70% to induce the secretion of TNF-alpha, IL-1 beta and IL-6 from normal blood mononuclear cells . This mechanism is likely to be operative in vivo, and could account for the protective effect of HA-1A, and its reduction of TNF-alpha production in vivo. Infection, 1993 Jul-Aug, 21(4), 245 - 7 The effect of clindamycin gel insert in periodontal pockets, as observed on smears and cultures; Sauvetre E et al.; This study is aimed at the evaluation of a 1% clindamycin hydrochloride containing gel on the microbial flora of periodontal pockets deeper than 5 mm . In order to achieve that purpose, 20 patients with pocketing in the premolar-molar regions were selected . Active and placebo gel were inserted once during the first 2 weeks of this experimental study . Microbial samplings were performed 1, 2, 4 and 12 weeks after the experiment started . The samples were submitted to microscopic examination and also to culture . Changes in the microbial content of the periodontal pockets treated by subgingival scaling and clindamycin 1% gel were significant, compared with those obtained with subgingival scaling and placebo gel, particularly with respect to anaerobic black-pigmented bacteria and the motile gram-negative flora . However, after 3 months, most of the treated cases were recolonized by the same initial species, though never at pre-clindamycin levels . In the light of this study, it will be concluded that the use of a small amount of clindamycin hydrochloride inserted into a periodontal pocket, once a week for 2 weeks as a complement to periodontal subgingival scaling, is beneficial in the treatment of adult periodontitis, by eliminating more effectively the microbial pocket colonization. Eur J Biochem, 1993 Jun 15, 214(3), 685 - 93 Outer-membrane porins from gram-negative bacteria stimulate platelet-activating-factor biosynthesis by cultured human endothelial cells; Tufano MA et al.; Porins are a family of hydrophobic proteins located in the outer membrane of the cell wall in Gram-negative bacteria . The effect of porins on the biosynthesis of platelet-activating factor (PAF) by cultured human umbilical-cord-vein-derived endothelial cells (HUVEC) was investigated . The results demonstrate that porins were able to induce a dose-dependent synthesis of PAF in HUVEC . PAF, synthesized after stimulation with porins, was mainly cell associated and the synthesis peaked at 15 min, decreasing rapidly thereafter . Experiments with radiolabeled precursors demonstrated that PAF, a 1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphorylcholine, was synthesized via the remodeling pathway involving the acetylation of 1-O-alkyl-2-lyso-sn-glyceryl-3-phosphorylcholine (2-lysoPAF) generated from 1-O-alkyl-2-acyl-sn-glyceryl-3-phosphorylcholine by phospholipase-A2 activity . The activation of phospholipase A2 in HUVEC stimulated by porins was detected by observing the mobilization of {14C}arachidonic acid . In addition, the activity of acetyl-CoA:1-alkyl-sn-glycero-3-phosphorylcholine 2-O-acetyltransferase was transiently increased in porin-stimulated HUVEC and, after incubation with {3H}CoASAc or {3H}acetate, the {3H}acetyl group was incorporated into newly synthesized PAF . Porins, by forming transmembrane channels, induced a sustained influx of extracellular 45Ca2+ into the cytosol . The activation of PAF synthesis by porins depended on this influx rather than on intracellular calcium mobilization, since PAF synthesis did not occur in the absence of extracellular Ca2+. J Immunol, 1993 Jun 15, 150(12), 5556 - 65 Neutrophil CD14: biochemical properties and role in the secretion of tumor necrosis factor-alpha in response to lipopolysaccharide; Haziot A et al.; CD14 is a myeloid cell differentiation Ag expressed primarily by monocytes and macrophages . CD14 has recently been shown to function as a receptor for a complex of LPS and LPS binding protein (LBP), an acute phase serum protein also present in normal serum in trace amounts . In the presence of LBP, LPS strongly activates monocytes via CD14 as measured by TNF secretion . This pathway of monocyte activation is thought to be a major contributor to the symptoms of endotoxin shock . Another major cell type involved in the response to Gram-negative infection is the neutrophil . Recent studies have shown that neutrophils also express CD14 and suggest that they can respond to LPS through a similar pathway . However, the biochemical nature of neutrophil CD14 has not previously been described . In this report, we have analyzed several biochemical characteristics of neutrophil CD14 . We show that CD14 is actively synthesized by neutrophils as a glycosylphosphatidyl-inositol-anchored protein, indistinguishable in size from monocyte CD14 . Furthermore, neutrophils, like monocytes, shed a smaller soluble form of CD14 into culture supernatants . In addition, like monocytes, neutrophils respond to LPS/LBP complexes via CD14 by releasing TNF-alpha . The described properties and function of neutrophil CD14 suggest that it may directly participate in the acute inflammatory response and in endotoxin shock. Schweiz Med Wochenschr, 1993 Jun 5, 123(22), 1165 - 8 {Cervical abscess caused by Capnocytophaga ochracea}; Henzen C et al.; Capnocytophaga is a gram-negative capnophilic bacterium which is part of the normal oral flora of humans (C . ochracea, C . gingivalis, C . sputigena) and mammals such as canines, cats, and rodents (C . animorsus and C . cynodegmi) . Its role in the pathogenesis of periodontal disease is not well defined, and normally it represents an opportunistic germ of low pathogenicity . Threatening and fulminant infections have been observed in immunodeficient patients, and lately in immunocompetent hosts . We describe an otherwise healthy woman who developed a cervical abscess due to C . ochracea . Recurrent aphthous lesions are suspected to be the port of entrance for the germs . Bacteriological, clinical, epidemiological, and therapeutic aspects of Capnocytophaga infection are discussed. Transplantation, 1993 Jun, 55(6), 1250 - 6 Lectin-dependent cell-mediated cytotoxicity and natural killer function in rejecting and infected lung allografts; Nguyen DM et al.; Differentiation between rejection and infection of lung allografts remains difficult . The effects of these two pathologic entities on the cytolytic activity of bronchoalveolar lavage (BAL) and PBL were investigated . Left lung allotransplantation was performed on 16 mongrel dogs of which 12 were available for complete studies . All animals received CA, AZA, and PRED for 2 weeks . Four grafts developed left lower lobe Gram negative pneumonia . The eight remaining recipients progressed gradually to severe rejection after acute reduction of immunosuppression . Cytolytic activity of blood and left lung BAL lymphocytes was quantitated by the natural killer (NK) and lectin-dependent cell-mediated cytotoxicity (LDCMC) assays . Two additional groups serving as controls were either given a 10-day course of immunosuppressants or had right lower lobe pneumonia induced by transbronchial inoculation of gram negative bacteria . Immunosuppressed control animals showed significant depression of PBL and BAL lymphocyte LDCMC and NK activity . Similarly, BAL lymphocytes expressed very low LDCMC in normal allografts (2.8 +/- 0.8%) . Once rejection developed and progressed, LDCMC became significantly higher (15.6 +/- 2.2 and 52.7 +/- 2.8% in mild and severe rejection, respectively) . There was no detectable NK activity in rejecting lung allografts . BAL lymphocytes from infected allografts, on the other hand, showed an elevation of both NK and LDCMC activity (9.1 +/- 1.1 and 14.6 +/- 1.0%, respectively) . Similarly, bacterial pneumonia in control animals manifested an increase in NK and LDCMC activity in lung and blood . PBL lymphocytes of lung allograft recipients, however, had increased NK and LDCMC activity in both rejection and infection . LDCMC/NK activity ratio (LM/NK index) of lung lymphocytes was significantly higher in rejecting allografts (11.2 +/- 1.0 and 12.4 +/- 1.6 for mild and severe rejection, respectively) than in infected ones (1.2 +/- 0.3, P < 0.0001) . It appears, from this study, that rejection of the lung allograft results in alterations in BAL lymphocyte phenotypes and functions that differ from those associated with bacterial infection . Such differences may be useful in distinguishing episodes of acute allograft rejection from bacterial infection. J Clin Invest, 1993 Jun, 91(6), 2850 - 60 Tissue factor pathway inhibitor reduces mortality from Escherichia coli septic shock; Creasey AA et al.; This study was designed to test the hypothesis that tissue factor pathway inhibitor (TFPI) plays a significant role in vivo in regulating coagulation that results from exposure of blood to tissue factor after vascular injury as in the case of gram negative sepsis . Highly purified recombinant TFPI (6 mg/kg) was administered either 30 min or 4 h after the start of a lethal intravenous Escherichia coli infusion in baboons . Early posttreatment of TFPI resulted in (a) permanent seven-day survivors (5/5) with significant improvement in quality of life, while the mean survival time for the controls (5/5) was 39.9 h (no survivors); and (b) significant attenuations of the coagulation response and various measures of cell injury, with significant reductions in pathology observed in E . coli sepsis target organs, including kidneys, adrenals, and lungs . TFPI administration did not affect the reduction in mean systemic arterial pressure, the increases in respiration and heart rate, or temperature changes associated with the bacterial infusion . TFPI treated E . coli infected baboons had significantly lower IL-6 levels than their phosphate buffered saline-treated controls, however tumor necrosis factor levels were similarly elevated in both groups . In contrast to the earlier 30-min treatment, the administration of TFPI at 4 h, i.e., 240 min, after the start of bacterial infusion resulted in prolongation of survival time, with 40% survival rate (2/5) and some attenuation of the coagulopathic response, especially in animals in which fibrinogen levels were above 10% of normal at the time of TFPI administration . Results provide evidence for the significance of tissue factor and tissue factor pathway inhibitor in bacterial sepsis, and suggest a role for blood coagulation in the regulation of the inflammatory response. Biophys J, 1993 Jun, 64(6), 1691 - 700 Deformations in the cytoplasmic membrane of Escherichia coli direct the synthesis of peptidoglycan . The hernia model; Norris V et al.; To explain the growth of the Gram-negative envelope and in particular how it could be strengthened where it is weakest, we propose in the hernia model that local weakening of the peptidoglycan sacculus allows turgor pressure to cause the envelope to bulge outwards in a hernia; the consequent local alteration in the radius of curvature of the cytoplasmic membrane causes local alterations in phospholipid structure and composition that determine both the synthesis and hydrolysis of peptidoglycan . This proposal is supported by evidence that phospholipid composition determines the activity of phospho-N-acetylmuramic acid pentapeptide translocase, UDP-N-acetylglucosamine:N-acetylmuramic acid-(pentapeptide)-P-P-bactoprenyl-N-acetylglucosamine transferase, bactoprenyl phosphate phosphokinase, and N-acetylmuramyl-L-alanine amidase . We also propose that the shape of Escherichia coli is maintained by contractile proteins acting at the hernia . Given the universal importance of membranes, these proposals have implications for the determination of shape in eukaryotic cells. J Clin Pharmacol, 1993 Jun, 33(6), 562 - 7 Determination of gentamicin pharmacokinetics by bioelectrical impedance in critically ill adults; Zarowitz BJ et al.; This investigation compares the accuracy of calculating gentamicin pharmacokinetic parameters by a noninvasive body composition technique (bioelectrical impedance analysis; BIA) with an empiric method, against the two-point method as the criterion standard . A prospective concurrent open label design was used . The 32 medical and surgical intensive care unit beds at Henry Ford Hospital, a not-for-profit, university-affiliated teaching hospital, served as the setting . Twenty critical ill adults, Therapeutic Index Scoring System (TISS) = 4, who required gentamicin as part of their normal course of therapy for gram-negative bacillary infections, were evaluated . Gentamicin Vd and k were calculated by three methods . After measurement of body composition parameters by BIA, previously derived gentamicin dosing equations were used to predict gentamicin volume of distribution (Vd) and elimination rate constant (k) (BIA method) . Empiric estimates of these parameters (Vd = 0.3L/kg and k derived from creatinine clearance) were compared with the BIA parameters against a criterion standard Vd and k determined from a two-point sampling of gentamicin serum concentrations . Measurements of BIA parameters and gentamicin serum concentrations were made in duplicate with coefficients of variation, < or = 2% and < or = 3%, respectively . The BIA and empiric methods produced resultant pharmacokinetic parameters (Vd and k) not different than those measured by the two-point method . There were no statistically significant differences in mean error (bias), or mean squared error (precision) for both Vd and k assessed by the empiric or BIA methods.(ABSTRACT TRUNCATED AT 250 WORDS) Anasthesiol Intensivmed Notfallmed Schmerzther, 1993 Jun, 28(4), 258 - 60 {Septic shock with acute abdomen in idiopathic hemochromatosis}; Sydow M et al.; The case of a patient with abdominal crisis and shock without any discernible origin who died 36 hours after hospital admission despite maximal therapy is described . Gram-negative sepsis, peritonitis and haemochromatosis with hepatic siderosis was the post-mortem diagnosis . We consider spontaneous peritonitis arising from translocation of normal intestinal flora (E . coli) through the intact wall of the gut combined with the impaired ability of the reticulo-endothelial system to remove endotoxin to be the causative factors . It is unknown whether the adrenal insufficiency due to siderophilic adrenal hypophysis and adrenal glands contributed to the fulminant course of the disease . Undiagnosed liver cirrhosis and especially haemochromatosis should therefore be included in the differential diagnostic considerations in patients presenting with these symptoms, and in whom no obvious cause for a septic focus can be found. Cesk Epidemiol Mikrobiol Imunol, 1993 Jun, 42(2), 87 - 92 {Lyme borreliosis: review of present knowledge}; Schwarzova K; The author reviews hitherto assembled knowledge on a bacterial disease, Lyme borreliosis transmitted by ticks . Initial information on Lyme borreliosis appeared at the beginning of the 20th century . In Czechoslovakia attention to the disease was paid since cca 1960 . The infection occurs as a rule in the summer months during the period when ticks are parasitic and at that time the causal agent of the disease is transmitted to hosts . Information on the prevalence and incidence of Lyme borreliosis in Europe is not complete and so far we do not possess a standardized diagnostic method for assessment of circulating antibodies in the patients' serum and cerebrospinal fluid . The infectious disease is caused by the gram-negative spirochete Borrelia burgdorferi . The Borrelia cell has a similar morphological structure as cells of other gram-negative bacteria . Chemical analysis of the external membrane of B . burgdorferi revealed the presence of 46% proteins, 51% lipids and 3% carbohydrates . The typical shape of borrelias indicates marked ondulation of 8-14 periplasmatic flagellae along the cell body . Borrelias can be cultivated in vitro in modified Barbour-Stoenner-Kelly medium at an optimal temperature of 30-37 degrees C . The change of morphology during cultivation is typical for B . burgdorferi . Clinically Lyme borreliosis is manifested in two stages . A typical manifestation of the early stage is a skin lesion--erythema migrans . The later stage is characterized above all by relapsing arthritis, CNS infection and chronic acrodermatitis chronica atrophicans . The disease is treated by administration of a number of antibiotics either by the oral route or by injection.(ABSTRACT TRUNCATED AT 250 WORDS) Changgeng Yi Xue Za Zhi, 1993 Jun, 16(2), 93 - 8 Ultrasonography and fine needle aspiration cytology of acute suppurative thyroiditis; Lin JD et al.; Although acute suppurative thyroiditis is a rare disorder, from January . 1985 to December 1991, we observed 11 cases of acute suppurative thyroiditis in our hospital . All patients underwent thyroid ultrasonography and fine needle aspiration biopsy and cytologic evaluation of the specimens . In eight cases, pus cultures were positive, four of them grew Gram negative bacteria . Ultrasonographic finding showed either local or diffuse low to a very low echo density on both lobes . However, a cystic lesion was also found . Polymorphonuclear cells were the main elements in the smears from the thyroid aspirates . In conclusion, thyroid ultrasound with fine needle aspiration and cytology can assist in confirming the early diagnosis of acute suppurative thyroiditis. Clin Infect Dis, 1993 Jun, 16 Suppl 4, S203 - 10 Bacterial mediators in periodontal disease; Loesche WJ; Periodontal disease is the general description given to the inflammatory response of the gingiva and underlying connective tissue to bacterial accumulations (dental plaque) on the teeth . A limited number of cultivable species are usually associated with periodontal disease . The majority of putative periodontal pathogens are gram-negative anaerobic rods . Some of the characteristics of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Treponema denticola will be discussed, given their prominence in the literature . These organisms share the ability to penetrate the gingival epithelium, such that their endotoxins, immunologically active compounds, and cytotoxic enzymes and molecules are presented directly to the host's inflammatory cells . This ability may be what distinguishes these gram-negative species from the plethora of other gram-negative species that inhabit the subgingival plaque . In addition, these organisms tend to be selected for in disease-associated plaques, suggesting that their nutritional needs are met when the gingival crevicular fluid contains a variety of inflammatory mediators and products of tissue breakdown . A . actinomycetemcomitans produces a leukotoxin, and the immunologic response of the host to this antigen may explain the unique pattern of tooth involvement in localized juvenile periodontitis . Both P . gingivalis and T . denticola have a trypsin-like enzyme that could be a virulence factor, primarily because this enzyme(s) may allow these organisms to grow in the presence of the inflammatory response of the host. Tokai J Exp Clin Med, 1993 Jun, 18(1-2), 71 - 80 Results and prognostic factors in the radical sinus operation for chronic sinusitis; Iida M et al.; Radical surgical procedures of the maxillary sinus with or without involvement of the ethmoid, sphenoid and frontal sinuses were performed in 382 adult patients in the Department of Otorhinolaryngology . Tokai University Hospital in the 8-year period from 1984 to 1991 . Of 382 patients, the postoperative symptomatology was studied in 189 patients by mcans of questionnaires . The postoperative results were finally analyzed in 63 patients whose subjective symptoms and objective findings were completely recorded . The follow-up period was over 3 months and subjective postoperative improvement was achieved in 83% and objective postoperative improvement in 75%; the overall improvement rate was 73% . In an analysis of results of the radical surgical operation, it appeared that poor results were closely related to the following factors: (1) maxillary sinus mucosa with cellular infiltration or edematous changes; (2) multiplicity of bacterial species including Gram-negative rods isolated from maxillary sinus secretion; (3) poor mucociliary clearance function; and (4) unsatisfactory postoperative treatment. J Immunol, 1993 Jun 1, 150(11), 5033 - 40 Differences in cytokine response and induction of nitric oxide synthase in endotoxin-resistant and endotoxin-sensitive mice after intravenous gram-negative infection; Evans TJ et al.; Previous reports have suggested that the endotoxin-resistant C3H/HeJ strain of mouse is more susceptible to infection than is the endotoxin-sensitive parent strain, C3H/HeN, although they have never been compared in an i.v . model of sepsis . We therefore have used these mouse strains in an i.v . model of Gram-negative sepsis to compare their sensitivities to infection, their cytokine responses, and the levels of induction of the enzyme nitric oxide synthase assayed in their livers . By using i.v . infection with Escherichia coli we have found that both strains are approximately equally sensitive to this organism, despite the C3H/HeJ mice having a markedly attenuated TNF-alpha response . IFN-gamma levels after infection were identical in the two strains; the levels of nitric oxide synthase induced in their livers were about fourfold greater in the C3H/HeJ mice . This difference could not be explained by differences in bacterial load . These experiments suggest that factors other than TNF-alpha are important in determining outcome from Gram-negative sepsis and that TNF-alpha is not a major factor in the induction of hepatic nitric oxide synthase after infection in vivo. FEBS Lett, 1993 May 17, 322(3), 231 - 4 Pentoxifylline inhibits the expression of tissue factor mRNA in endotoxin-activated human monocytes; Ollivier V et al.; Tissue factor (TF) is a transmembrane glycoprotein which, in association with factor VII(a), is the main activator of coagulation . In illnesses such as Gram-negative endotoxemia, circulating monocytes synthesize and express substantial TF activity, resulting in extensive disseminated intravascular coagulation . We investigated the way in which TF is suppressed by pentoxifylline (PTX), and found that PTX down-regulates immunologic TF expression and specific mRNA production in response to LPS . Since TF mRNA stability is not altered, this effect appears to take place at the transcriptional level. Gene, 1993 May 15, 127(1), 127 - 31 Sequence and transcriptional analysis of the nourseothricin acetyltransferase-encoding gene nat1 from Streptomyces noursei; Krugel H et al.; We have determined the nucleotide (nt) sequence of nat1, a gene encoding nourseothricin (Nc) acetyltransferase (AT) from Streptomyces noursei, and its transcriptional start point (tsp) . The nt sequence upstream from the coding region is completely different from that of the stat gene (encoding streptothricin AT) from Streptomyces lavendulae {S . Horinouchi, K . Furuya, M . Nishiyama, H . Suzuki and T . Beppu, J . Bacteriol . 169 (1987) 1929-1937}, even though the nt sequences of the two genes and the deduced amino acid (aa) sequences of the two enzymes show a high degree of similarity . Another stat gene, derived from a Gram-negative plasmid, showed only deduced aa similarity, but not nt sequence similarity, to the above two . A database search for related aa sequences did not reveal any clear-cut homologies to other types of protein . A multiple aa sequence alignment of several ATs is presented. JAMA, 1993 May 5, 269(17), 2221 - 7 A controlled trial of HA-1A in a canine model of gram-negative septic shock; Quezado ZM et al.; OBJECTIVE--To investigate the therapeutic efficacy and microbiological and physiological effects of a human IgM monoclonal antibody (HA-1A) directed against the lipid A component of endotoxin in a canine model of sepsis that simulates the cardiovascular abnormalities of human septic shock . DESIGN--Blinded, placebo-controlled 28-day trial . INTERVENTIONS--Purpose-bred beagles were implanted with an intraperitoneal clot infected with Escherichia coli O111:B4 . At clot placement, animals received HA-1A (10 mg.kg-1), control human IgM antibody (10 mg.kg-1), or control human serum albumin intravenously . All animals were given antibiotic and fluid therapy . MEASURES--Survival and microbiological and physiological events . RESULTS--Only two (15%) of 13 animals in the HA-1A group, compared with eight (57%) of 14 control animals (combined control human IgM antibody and control human serum albumin groups) (P = .05), survived 28 days . At 24 hours, the HA-1A group had lower mean arterial pressure (P = .04) and cardiac index (P = .004) and higher lactate levels (P = .05) compared with the combined-controls group . In addition, these parameters in the HA-1A group were significantly more predictive of death . The HA-1A and combined-controls groups had similar significant increases in the level of endotoxemia and bacteremia . Studies of toxic effects showed no harmful effects of control human IgM antibody in infected animals or HA-1A in non-infected animals . CONCLUSION--In a canine model of E coli sepsis, HA-1A did not alter levels of bacteremia or endotoxemia and actually decreased survival . If these data are relevant to human septic shock, HA-1A therapy should be limited until the conditions under which this monoclonal antibody has beneficial or deleterious effects are more completely defined. Biochemistry, 1993 May 4, 32(17), 4579 - 86 Effect of pH on solubility and ionic state of lipopolysaccharide obtained from the deep rough mutant of Escherichia coli; Din ZZ et al.; The dissociation of the highly aggregated form of lipopolysaccharide (LPS) from Gram-negative bacteria to the monomeric (or soluble) form is though to be the initial step in the activation of responding cells (macrophages, B-cells, neutrophils, monocytes, and endothelial cells) by LPS . This process is presently not adequately understood . Using the equilibrium dialysis apparatus and a highly purified and well-characterized radiolabeled deep rough chemotype LPS ({14C}ReLPS) from Escherichia coli D31m4, we have examined the effect of pH on its solubility (CT) and ionic states in aqueous media . The solubility range of {14C}ReLPS suspended in 50 mM Tris-HCl-100 mM KCl buffer (or 50 mM MES-100 mM KCl buffer at pH 6.5) was determined to be from (2.91 +/- 0.01) x 10(-8) to (4.55 +/- 0.07) x 10(-8) M over a pH range of 6.50-8.20, respectively . These experimental data satisfactorily fitted the curve generated by the solubility equation CT = S0(1 + K5/{H+})/({H+}/K4' + 1), where S0 is the concentration of the tetraanionic ReLPS, K5 is the dissociation constant of the tetraanionic ReLPS in solution, and K4' is the dissociation constant of the trianionic ReLPS at the surface of the solid particles in suspension . The increase in solubility of ReLPS with increase in pH from 7.00 to 8.20 is primarily caused by the formation of the pentaanionic form from the tetraanions . The pK5 (primarily the second dissociation of the 1-phosphate) of ReLPS was determined to be 8.58 from experimental data.(ABSTRACT TRUNCATED AT 250 WORDS) Toxicol Lett, 1993 May, 68(1-2), 251 - 63 Ozone- and endotoxin-induced mucous cell metaplasias in rat airway epithelium: novel animal models to study toxicant-induced epithelial transformation in airways; Harkema JR et al.; Mucous (goblet) cell proliferation and hypersecretion of airway mucus are important characteristics of human respiratory disorders, especially chronic bronchitis and cystic fibrosis . These changes in secretory patterns also occur in animals experimentally exposed to chemical irritants such as ozone (O3), sulfur dioxide (SO2), and cigarette smoke . The cellular and molecular mechanisms involved in irritant-induced mucous cell metaplasia (MCM; transformation of airway epithelium, normally devoid of mucous cells, to a secretory epithelium containing numerous mucous cells) are still unclear . We used two experimental models of toxicant-induced MCM in rat airways to study the cellular and molecular changes that occur during the development of this respiratory tract lesion . MCM can be induced in the nasal transitional epithelium of rats by repeated exposure to ambient levels of ozone . In addition, MCM can be induced in the tracheobronchial airways of rats repeatedly exposed to endotoxin, a lipopolysaccharide-protein molecule found in the outer walls of Gram-negative bacteria . The pathogenesis of ozone- or endotoxin-induced MCM has been partially characterized using a variety of morphometric and histochemical techniques . Toxicant-induced changes in the numbers and types of airway epithelial cells have been estimated using morphometric methods designed for estimating the abundance of cell populations . Nasal pulmonary airway tissues are also processed for light microscopy and stained with Alcian Blue (pH 2.5)/Periodic Acid Schiff (AB/PAS) for detection of acidic and neutral mucosubstances (the specific glycoprotein product of mucous cells), respectively, within the tissue . Computerized image analysis is used to quantitate the amount of the stained mucous product within the airway epithelium . To better characterize the molecular and cellular events in the pathogenesis of ozone- or endotoxin-induced MCM in the rat airway epithelium, we are conducting studies to determine when, and in which epithelial cells, the mucin gene is expressed after exposure to the toxicant . In these studies, rats undergo single or repeated exposures to ozone or endotoxin and are then sacrificed immediately or a few days after the end of the exposures . Airway tissues are microdissected from specific regions of the exposed respiratory tract, and changes in mucin core polypeptide mRNA are evaluated by Northern analysis using human and rat mucin cDNA . In future studies using in situ hybridization, we will establish when, and in which epithelial cells, the expression of high molecular weight airway mucin is initiated in response to ozone or endotoxin.(ABSTRACT TRUNCATED AT 400 WORDS) J Clin Microbiol, 1993 May, 31(5), 1122 - 6 Occurrence of bacterial endosymbionts in Acanthamoeba spp . isolated from corneal and environmental specimens and contact lenses; Fritsche TR et al.; Free-living and parasitic protozoa are known to harbor a variety of endosymbiotic bacteria, although the roles such endosymbionts play in host survival, infectivity, and invasiveness are unclear . We have identified the presence of intracellular bacteria in 14 of 57 (24%) axenically grown Acanthamoeba isolates examined . These organisms are gram negative and non-acid fast, and they cannot be cultured by routine methodologies, although electron microscopy reveals evidence for multiplication within the amoebic cytoplasm . Examination for Legionella spp . with culture and nucleic acid probes has proven unsuccessful . We conclude that these bacteria are endosymbionts which have an obligate need to multiply within their amoebic hosts . Rod-shaped bacteria were identified in 5 of 23 clinical Acanthamoeba isolates (3 of 19 corneal isolates and 2 of 4 contact lens isolates), 4 of 25 environmental Acanthamoeba isolates, and 2 of 9 American Type Culture Collection Acanthamoeba isolates (ATCC 30868 and ATCC 30871) previously unrecognized as having endosymbionts . Coccus-shaped bacteria were present in one clinical (corneal) isolate and two environmental isolates . There was no statistical difference (P > 0.8) between the numbers of endosymbiont strains originating from clinical (26% positive) and environmental (24% positive) amoebic isolates, suggesting that the presence alone of these bacteria does not enhance amoebic infectivity . Rods and cocci were found in both clinical and environmental isolates from different geographical areas (Seattle, Wash., and Portland, Oreg.), demonstrating their widespread occurrence in nature . Our findings suggest that endosymbiosis occurs commonly among members of the family Acanthamoebidae and that the endosymbionts comprise a diverse taxonomic assemblage . The role such endosymbionts may play in pathogenesis remains unknown, although a variety of exogenous bacteria have been implicated in the development of amoebic keratitis, warranting further evaluation. Cell Immunol, 1993 May, 148(2), 397 - 407 Iron augments macrophage-mediated killing of Brucella abortus alone and in conjunction with interferon-gamma; Jiang X et al.; Brucella abortus are Gram negative facultative intracellular bacteria, which survive and replicate in host macrophages . We have recently demonstrated that activation of macrophages with interferon-gamma increases their anti-brucella activities but does not result in elimination of intracellular brucellae . Here we demonstrate that iron-loaded macrophages have an enhanced capacity to kill or prevent replication of intracellular brucellae . Iron added bound to transferrin or as a salt, iron-nitrilotriacetate, can mediate the effect . Macrophages supplemented with iron-loaded transferrin in addition to activation with interferon-gamma can frequently eliminate the intracellular organisms by 48 hr after infection . The effect is apparent following phagocytosis of either nonopsonized or antibody-opsonized brucellae, and with both attenuated and virulent strains of B . abortus . The killing can be blocked by the hydroxyl radical scavengers mannitol and thiourea . This is consistent with the Haber-Weiss reaction, in which iron catalyzes the generation of hydroxyl radicals from hydrogen peroxide. Infect Immun, 1993 May, 61(5), 1756 - 63 Reactivity of the human antiendotoxin immunoglobulin M monoclonal antibody HA-1A with lipopolysaccharides from rough and smooth gram-negative organisms; Mascelli MA et al.; Clinical data suggest that the human immunoglobulin M antiendotoxin antibody HA-1A reduced mortality in patients diagnosed with gram-negative bacteremia and bacteremia with shock . Previous studies have demonstrated that HA-1A binds to the lipid A domain of lipopolysaccharide (LPS) . The present study evaluated the ability of HA-1A to interact with LPs isolated from various strains of gram-negative bacteria by using liquid-phase rate nephelometry and solid-phase immunoblotting assays . HA-1A formed immune complexes in solution with LPSs isolated from both rough and smooth gram-negative organisms . Western blot (immunoblot) analysis of these LPS preparations revealed that HA-1A bound to LPS isolated from rough gram-negative organisms and to a rough LPS-like component present in smooth LPS . HA-1A also bound to LPS-protein complexes found in certain commercial rough LPS preparations . Preincubation of HA-1A with lipid A completely blocked subsequent binding of HA-1A to LPS in both liquid- and solid-phase assay formats, suggesting that the interaction of HA-1A with LPS is through the lipid A domain . Evidence that the binding of HA-1A to LPS was mediated through the antigen-combining (Fv) region of the antibody was provided by the finding that a murine anti-idiotypic antibody to HA-1A inhibited binding . These findings suggested that the broad antiendotoxin reactivity exhibited by HA-1A appeared to be due to the ability of HA-1A to bind to the conserved lipid A moiety of LPSs derived from both smooth- and rough-phenotype gram-negative bacterial strains. Infect Immun, 1993 May, 61(5), 1715 - 21 Cytocidal effects of Escherichia coli hemolysin on human T lymphocytes; Jonas D et al.; Escherichia coli hemolysin is the prototype of a large family of pore-forming toxins produced by gram-negative organisms . Besides its known cytotoxic activities against granulocytes, monocytes, endothelial cells, and renal epithelial cells, we now demonstrate that the toxin potently kills human T lymphocytes . Evidence based on different and independent approaches indicates that lymphocidal activity is due to formation of transmembrane pores . Additionally, cells prestimulated with phytohemagglutinin respond to low doses of E . coli hemolysin with DNA fragmentation similar to that observed in cells undergoing programmed cell death . Kinetic considerations lead us to conclude that DNA degradation may, however, represent an epiphenomenon . Killing of T cells is another means through which E . coli hemolysin could directly impair host defense. Infect Immun, 1993 May, 61(5), 1630 - 5 Endotoxin-induced tumor necrosis factor alpha synthesis in murine embryo fibroblasts; Havell EA et al.; Murine embryo fibroblasts (MEF) were found to secrete tumor necrosis factor (TNF) in response to stimulation with endotoxin . Endotoxin-induced TNF production by MEF was inhibited by cycloheximide . However, reversal of the effect of this inhibitor on protein synthesis results in TNF being secreted in amounts equivalent to those produced by endotoxin-induced MEF not treated with cycloheximide . Actinomycin D treatment of MEF blocked the production of endotoxin-induced TNF . Maximal production of TNF required MEF gene transcription during the first 6 h of incubation with endotoxin . To determine whether endotoxin-induced TNF alpha (TNF-alpha) and/or TNF beta were produced by MEF, cDNA was synthesized from the total RNA isolated from endotoxin-induced MEF and amplified by the polymerase chain reaction in the presence of oligonucleotide primers specific for each cytokine . On the basis of the polymerase chain reaction analysis, it was determined that TNF-alpha mRNA levels were increased in endotoxin-induced MEF . Thus, production of TNF-alpha by fibroblasts in response to the endotoxin component of bacterial cell walls is likely to contribute to the expression of TNF-mediated effects occurring in fibroblast-rich tissues infected with gram-negative bacteria. Am J Crit Care, 1993 May, 2(3), 224 - 35; quiz 236-7 Epidemiology, pathophysiology and clinical presentation of gram-negative sepsis; Hazinski MF et al.; OBJECTIVE: To review the epidemiology and pathophysiology of gram-negative sepsis and the new consensus terminology describing the clinical signs of sepsis . DATA SOURCES: Review of the medical literature and compiled data from animal and clinical trials . PARTICIPANTS: Members of the Society of Critical Care Medicine, American College of Chest Physicians and American Association of Critical-Care Nurses with expertise on the subject of sepsis and its complications . RESULTS: Preconference and general sessions were offered at the National Teaching Institutes of the American Association of Critical-Care Nurses, with the goal of clarifying the epidemiology, risk factors and pathophysiology of gram-negative sepsis . In addition, current terminology and new (1992) consensus terminology describing the clinical signs of sepsis were presented . Special emphasis was placed on the role of the healthcare provider in the prevention and recognition of sepsis and the role of the septic mediators in the septic cascade . CONCLUSIONS: If the incidence of sepsis is to be reduced, the healthcare provider must be aware of the risk factors for sepsis and methods of reducing nosocomial infections . A thorough understanding of the role of mediators and consensus terminology used to describe sepsis, severe sepsis, septic shock and multiple organ dysfunction syndrome is necessary to recognize early or progressive signs of sepsis and to initiate state-of-the-art therapy. J Gen Microbiol, 1993 May, 139 ( Pt 5), 1105 - 14 The melA gene is essential for melanin biosynthesis in the marine bacterium Shewanella colwelliana; Fuqua WC et al.; The surface-adhering, Gram-negative marine bacterium Shewanella colwelliana synthesizes a red-brow melanin in the late stage of exponential growth in laboratory culture . Previous studies identified a single gene, melA, from S . colwelliana that could impart the ability to produce melanin to an E . coli host . However, these studies did not demonstrate a requirement for melA during melanization in S . colwelliana . In this paper, genetic analyses, using a broad host range conjugation system to generate specific lesions, reveal that melA null mutants fail to synthesize pigment . The wild-type melA gene provided in trans on a low copy number plasmid complemented these null mutations, as well as a spontaneous pigment variant, to wild-type melanin synthesis . Polyclonal antibodies, raised against a MelA-LacZ fusion protein, were used to confirm the presence of the melA gene product in wild-type S . colwelliana and verify its absence in the non-pigmented mutants . In addition, detection of the MelA protein over the course of growth in batch culture revealed a constant steady-state level of MelA protein, suggesting that the timing of melanization and the quantity of melanin synthesized is not controlled at the level of melA expression. J Antimicrob Chemother, 1993 May, 31 Suppl D, 61 - 70 Pharmacodynamics of antibiotics in the therapy of meningitis: infection model observations; Schmidt T et al.; Detailed studies of pharmacodynamic principles relevant to the therapy of bacterial meningitis are difficult to perform in man, while the rabbit model of bacterial meningitis has proved to be extremely valuable and has led to insights that appear relevant for the treatment of humans . Most importantly in the light of the restricted penetration of antibiotics into the CSF, animal studies have shown that in meningitis there is a dose-response curve between the CSF concentrations achieved by antibiotics and their bactericidal activity . This appears to be true for all classes of antibiotics thus far examined, including the beta-lactams, which do not show such a dose-response behaviour in other infections . Only CSF concentrations that exceed the MBC of the infecting organism by at least 10-30-fold achieve consistent and rapid bactericidal activity . Such rapid bactericidal activity is a requirement for successful therapy with beta-lactams and can be impaired with certain antibiotics by the specific conditions in infected CSF (protein content; acidic pH; slow-growing bacteria) . However, rapid antibiotic killing of the infecting organisms may not be without adverse effects either . Some antibiotics, particularly beta-lactams lead to the brisk liberation of bacterial cell wall components (e.g . endotoxin, in the case of Gram-negative organisms) which have an inflammatory effect on the host and can lead to a temporary deterioration of the disease . Dexamethasone, when administered with the antibiotic, can prevent some of the adverse effects of rapid bacterial lysis. Circ Shock, 1993 May, 40(1), 53 - 60 Lipopolysaccharide alters suckling rat liver glycogenolysis; Goto M et al.; Gram-negative sepsis/septic shock in the newborn continues to be a major medical problem, causing high mortality . Hyperglycemia followed by hypoglycemia is a common symptom in endotoxic shock . However, the mechanism of newborn glucoregulatory response to endotoxin has not been well understood . Paradoxically, monocyte-phagocytes can contribute to shock by overwhelming secretion of cytokines and also host defense by detoxifying endotoxin . Since monocyte-phagocyte function is immature in the newborn, this study was performed to evaluate Kupffer cell's role in liver glycogenolysis during endotoxic shock . Endotoxin (LPS) induced hyperglycemia in 10-day-old rats, and increased net glucose output in the isolated perfused liver . 1) Cytarabine decreased Kupffer cell function (decreased hepatic colloid carbon uptake) and blunted LPS-increased liver net glucose output in the Cytarabine + LPS-treated group (104 +/- 4 vs . 146 +/- 3 micrograms/min/g wet liver in the LPS-treated group: P < .001) . 2) Indomethacin (IND) suppressed LPS-induced liver net glucose output in the LPS + IND-treated group (133 +/- 5 vs . 146 +/- 3 micrograms/min/g wet liver, P < .05) . Thus, prostaglandins were suggested to contribute to glycogenolysis in the 10-day-old rat liver . 3) Phorbol 12-myristate 13-acetate (PMA) increased liver net glucose output (166 +/- 4 micrograms/min/g wet liver), and H-7, a protein kinase C inhibitor, blunted PMA-induced liver glucose output (140 +/- 2 micrograms/min/g wet liver, P < .05) . H-7 enhanced LPS-induced liver net glucose output (196 +/- 9 micrograms/min/g wet liver, P < .01) . Therefore, protein kinase C may not be the dominant cell signaling system for LPS stimulation in suckling rat Kupffer cells. J Dairy Res, 1993 May, 60(2), 223 - 8 Routine limulus amoebocyte lysate (LAL) test for endotoxin determination in milk using a Toxinometer ET-201; Mottar J et al.; A rapid method of performing the Limulus amoebocyte lysate (LAL) test in milk is proposed using the Toxinometer ET-201 . This instrument measured the increase in turbidity due to the interaction between the endotoxins of the Gram-negative bacteria and the LAL reagent, monitored the ratio Rt of the sequential to the initial transmission at 12 s intervals and quantified endotoxins by determination of the reaction time Tr required to obtain a 5% decrease in Rt . There was a good correlation between the toxinometrically determined endotoxin concentrations and the number of Gram-negative bacteria (SD, 0.18 log(plate count units)), and the repeatability (CV, 6-10%) was high . The assay may be useful for screening raw materials for UHT milk production, as the endotoxin content of the raw material is related to the rest proteinase activity in the UHT milk. Lancet, 1993 May 1, 341(8853), 1133 - 5 Reappraisal of the role of endotoxin in the sepsis syndrome; Hurley JC; There is strong evidence to implicate endotoxin released from gram negative bacteria in the pathogenesis of the sepsis syndrome and related conditions, but equally compelling data bring the role of endotoxin into doubt . Reappraisal of endotoxin and its release from gram negative bacteria suggests that it is not directly responsible for the complications of sepsis syndrome . Rather, release of endotoxin is a marker for the transition of gram negative organisms to cell-wall-deficient forms (L-forms) that may persist undetected despite antibiotic therapy directed against the parental form . This transition has two consequences in compromised patients: L-forms cause organ failure, and they serve as a sanctuary from which cell-wall-intact revertants may arise. New Horiz, 1993 May, 1(2), 312 - 23 Disseminated intravascular coagulation: pathophysiology, diagnosis, and treatment; ten Cate H et al.; Disseminated intravascular coagulation is a frequent finding in critically ill patients, and may be diagnosed in the majority of patients with Gram-negative sepsis . Tissue damage may result from intravascular thrombosis, and disseminated intravascular coagulation is an underestimated causal factor in the pathogenesis of organ failure in sepsis . The diagnosis of disseminated intravascular coagulation is difficult, as the initial coagulation process that leads to thrombosis is counteracted by fibrinolytic responses, that in the context of ongoing consumption of clotting factors may result in an overwhelming bleeding tendency . Hence, depending on underlying disease, and the time at which the patient is evaluated, different laboratory results may be obtained . The treatment of disseminated intravascular coagulation is even more challenging than its diagnosis . No well-designed, controlled clinical trials exist that form a basis for rational treatment decisions . Treatment frequently needs to be individualized, and rapid adjustments may be necessitated by the course of the disease . Nonetheless, we believe that recent insights in the pathophysiology of disseminated intravascular coagulation, in particular concerning the role of the extrinsic coagulation pathway, provide ground for some optimism concerning future therapeutic options. Trends Microbiol, 1993 May, 1(2), 50 - 5 A novel secretion apparatus for the assembly of adhesive bacterial pili; Jacob-Dubuisson F et al.; The biogenesis of most types of bacterial pili requires two specialized proteins: a chaperone that caps the pilus subunits in the periplasm, and an outer membrane usher that receives the subunits and serves as an assembly platform . This secretion and assembly machinery is proposed to be a novel export apparatus found widely in Gram-negative pathogens. J Struct Biol, 1993 May-Jun, 110(3), 180 - 7 Demonstration of the glycocalyces associated with three oral gram-negative bacterial species using a modern acrylic resin technique; Barber PM et al.; The complex highly hydrated chemical composition of the bacterial glycocalyx renders it difficult to preserve and visualize at the ultrastructural level . Polyanionic stains such as ruthenium red help to maintain some structural integrity, and other more modern approaches include antibody stabilization, lectins, and the addition of lysine to the primary fixative . It has been suggested that the glycocalyx of certain disease-associated organisms may play a role in the pathogenesis of some microbially based diseases such as periodontitis . New, more adequate, modern methodologies are therefore required for the further study of this structure . In the present study a cold dehydration process in conjunction with LR white acrylic resin has been employed to study the glycocalyces of three oral gram-negative bacterial species reported to be periodontopathogens . When compared with organisms processed conventionally and with ruthenium red, the organisms processed by the cold dehydration and LR white method demonstrated a fibrous matrix that was not seen in the other specimens . These results indicate that a combination of reduced dehydration temperature and cold acrylic resin embedding provides the best methodology for the visualization of the fine structure of the bacterial glycocalyx . This approach may be particularly useful in the study of organisms within specific disease-associated environments such as the periodontal pocket. J Clin Microbiol, 1993 May, 31(5), 1027 - 9 Rejection criteria for endotracheal aspirates from adults; Morris AJ et al.; Although criteria have been established to assess the quality of sputum specimens, no criteria for assessing the quality of endotracheal suction aspirates (ETSA) exist . Therefore, we compared the Gram stain (GS) and culture results for 504 consecutive ETSA specimens . Results recorded for GS included the numbers of squamous epithelial cells (SEC) and polymorphonuclear leukocytes (PML) per low-power field (LPF) (magnification, x100) as well as the quantities and types of organisms per high-power field (HPF) (magnification, x1,000) . Culture results were quantitated by organism . Only 15% of ETSA specimens tested by GS contained > 10 SEC per LPF, and 21, 20, and 59% had < or = 10, 11 to 24, and > or = 25 PML per LPF, respectively . For 40% of ETSA specimens, no organisms were visible by GS . Of these specimens, 40% were sterile, 48% grew normal oropharyngeal flora (NF) only, 5% grew 1+ NF (i.e., > 10 colonies in the first quadrant) and 1+ gram-negative rods (GNR), and 7% grew < or = 1+ GNR either alone or in mixed culture . The mean numbers of organisms recovered from ETSA with < or = 10 SEC per LPF and > 10 SEC per LPF were 2.35 and 4.05, respectively . We therefore recommend that ETSA specimens that show no organisms by GS be rejected, in addition to those with > 10 SEC per LPF . Application of these rejection criteria enabled us to reject 847 (41%) of 2,068 ETSA specimens over a 6-month period . This represents a saving of approximately $66,000/year in unnecessary laboratory charges to patients. J Clin Invest, 1993 May, 91(5), 2076 - 83 Mechanisms of stimulation of interleukin-1 beta and tumor necrosis factor-alpha by Mycobacterium tuberculosis components; Zhang Y et al.; The granulomatous immune response in tuberculosis is characterized by delayed hypersensitivity and is mediated by various cytokines released by the stimulated mononuclear phagocytes, including tumor necrosis factor-alpha (TNF alpha) and IL-1 beta . We have demonstrated that Mycobacterium tuberculosis cell wall component lipoarabinomannan (LAM), mycobacterial heat shock protein-65 kD, and M . tuberculosis culture filtrate, devoid of LPS as assessed by the Amebocyte Lysate assay, stimulate the production of TNF alpha and IL-1 beta proteins and mRNA from mononuclear phagocytes (THP-1 cells) . The effect of LAM on the release of these cytokines was specific, as only LAM stimulation was inhibited by anti-LAM monoclonal antibody . Interestingly, we found that LAM and Gram-negative bacterial cell wall-associated endotoxin LPS may share a similar mechanism in their stimulatory action as demonstrated by inhibition of TNF alpha and IL-1 beta release by monoclonal antibodies to CD14 . Anti-CD14 monoclonal antibody MY4 inhibited both TNF alpha and IL-1 beta release with LAM and LPS but no effect was observed with other mycobacterial proteins . An isotype antibody control did not inhibit release of cytokines under the same experimental conditions . M . tuberculosis and its components upregulated IL-1 beta and TNF alpha mRNAs in THP-1 cells . Nuclear run-on assay for IL-1 beta demonstrated that LAM increased the transcription rate . The induction of IL-1 beta was regulated at the transcriptional level, in which these stimuli acted through cis-acting element(s) on the 5' flanking region of the IL-1 beta genomic DNA . M . tuberculosis cell wall component LAM acts similarly to LPS in activating mononuclear phagocyte cytokine TNF alpha and IL-1 beta release through CD14 and synthesis at the transcriptional level; both cytokines are key participants in the host immune response to tuberculosis. J Immunol, 1993 Apr 15, 150(8 Pt 1), 3397 - 403 Mice treated with a leumedin or antibody to Mac-1 to inhibit leukocyte sequestration survive endotoxin challenge; Burch RM et al.; Endotoxin challenge causes metabolic dysfunction mediated by TNF, and sequestration of leukocytes . NPC 15669, N-carboxy-L-leucine, N-{2,7-dimethylfluoren-9-yl)methyl} ester, inhibits leukocyte recruitment into inflammatory lesions in animals, and inhibits endotoxin-induced neutropenia and lymphopenia in mice . This study was carried out to determine whether the ability of NPC 15669 to inhibit leukocyte sequestration is sufficient to promote survival after endotoxin challenge . To inhibit leukocyte sequestration directly, mice were treated with anti-CD11a (LFA-1) or anti-CD11b (Mac-1) before endotoxin challenge . Anti-CD11b partly inhibited neutropenia and lymphopenia in response to challenge with LPS, but anti--CD11a had little effect on leukopenia . At doses of 100 and 1000 micrograms/kg, anti-CD11b increased survival to endotoxin challenge from 0 to 20 and 40%, respectively, whereas anti-CD11a was without effect . These observations, coupled with the finding that NPC 15669 does not inhibit endotoxin-induced TNF release suggest that inhibition of leukocyte sequestration can increase survival after endotoxin challenge, and that NPC 15669 or antibodies to Mac-1 may represent effective therapies for gram-negative sepsis and shock. JAMA, 1993 Apr 14, 269(14), 1829 - 35 Anticytokine strategies in the treatment of the systemic inflammatory response syndrome; Dinarello CA et al.; The systemic inflammatory response syndrome (SIRS) is an acute illness characterized by generalized activation of the endothelium . The most severe form of the syndrome is found in patients with shock due to gram-negative sepsis . We examined both animal and limited human data for the contribution of cytokines to this syndrome . Cytokines are endogenously produced proteins of small molecular weight and multiple biological effects . The cytokines interleukin 1 (IL-1) and tumor necrosis factor (TNF), as well as interferon-gamma and interleukin 8, are discussed . Laboratory investigations suggest that these cytokines play a critical role in SIRS by promoting the biochemical and clinical characteristics of SIRS . The biochemical changes induced by TNF and IL-1 include increased synthesis of nitric oxide, prostaglandins, platelet-activating factor, and endothelial cell adhesion molecules . Specific blockade of TNF using neutralizing antibodies or soluble receptors to TNF in animal models of SIRS reduces mortality and severity of disease . Similar results have been observed blocking IL-1 using soluble IL-1 receptors or IL-1 receptor antagonists . Preliminary clinical studies suggest that blockade may be useful in treating human SIRS . The various strategies for blocking IL-1 and TNF are presented; in addition, their mechanism(s) of action and safety in humans are discussed . We conclude that based on animal studies and preliminary clinical trials, strategies to block IL-1 or TNF may benefit patients with the syndrome, although thorough clinical trials have not been completed. Anaesth Intensive Care, 1993 Apr, 21(2), 172 - 3 Aminoglycoside volume of distribution and illness severity in critically ill septic patients; Marik PE; The volume of distribution of amikacin and the APACHE II score were determined in 42 critically ill patients being treated for a gram-negative infection . The mean volume of distribution (Vdt) was 0.41 +/- 0.12 l/kg with a wide range (normal of 0.25 l/kg) . There was a good relationship between the Vdt and illness severity as measured by the APACHE II score (r = 0.70; P < 0.001) . Critically ill patients should receive larger loading doses of aminoglycosides in order to achieve therapeutic blood levels . The aminoglycoside Vdt may be useful in determining the degree of capillary leak and tissue oedema that accompanies sepsis. Infusionsther Transfusionsmed, 1993 Apr, 20 Suppl 1, 16 - 9; discussion 20 {Detection of endotoxin in plasma: specificity and value for development and prognosis of infection}; Urbaschek R et al.; A number of problems may be involved in the detection of endotoxin in plasma of patients using LAL (Limulus amebocyte lysate) . When collecting blood or processing samples, contamination with endotoxin or its adsorption to material must be avoided . In our laboratory a kinetic LAL microtiter assay was developed that takes into account plasma-related interferences with the LAL endotoxin reaction by performing an internal standardization in each sample . Negative results do not absolutely exclude the involvement of endotoxins in the underlying disease . High levels of endotoxins do not necessarily reflect the severeness of the clinical status of the patient . Due to nonendotoxin-specific reactions with some complete lysates, false-positive levels may result, e.g., following immunoglobulin therapy . In spite of these limitations, the LAL test remains a valuable tool in the evaluation of gram-negative infections. Am J Vet Res, 1993 Apr, 54(4), 576 - 9 Pharmacokinetics of cefotaxime in neonatal pony foals; Gardner SY et al.; Serum concentrations of cefotaxime and desacetylcefotaxime were measured in 1-week-old pony foals after IV administration of a single dose of cefotaxime . The cefotaxime disposition data conformed to a two-compartment model with elimination half-life of 0.60 hour . The combined cefotaxime and desacetylcefotaxime data was best described by a four-compartment model . The apparent half-life describing the disappearance of desacetylcefotaxime was 1.69 hours . Dosage of 40 mg/kg of body weight given IV every 4 to 6 hours for neonatal foals with gram-negative septicemia and every 2 hours for foals with meningitis is recommended for further study. J Am Vet Med Assoc, 1993 Apr 1, 202(7), 1137 - 42 Cholelithiasis in dogs: 29 cases (1980-1990); Kirpensteijn J et al.; Medical records of 29 dogs with cholelithiasis were reviewed . Aged female small-breed dogs were overrepresented . Mean age was 9.5 years, and mean weight was 12 kg . Vomiting, anorexia, weakness, polyuria/polydipsia, weight loss, icterus, fever, and signs of abdominal pain were the most common clinical signs . Leukocytosis, neutrophilia with left shift, monocytosis, high activity of serum hepatic enzymes, hypoalbuminemia, and high concentrations of serum total bilirubin were common . Radiopaque choleliths were evident on abdominal radiography of 13 of 27 dogs . Microbial culturing of bile isolated organisms in 15 of 20 dogs . Gram-negative bacteria were most common . Surgery was performed in 22 dogs . Four dogs were treated medically, and 3 dogs were euthanatized without treatment . Surgical treatment consisted of cholecystectomy in 11 dogs, choledochotomy in 5 dogs, cholecystotomy in 4 dogs, and cholecystojejunostomy in 1 dog . Sphincter of Oddiotomy was performed in 1 dog . Five dogs had concurrent generalized peritonitis attributable to bile . Multiple choleliths were detected in most of the dogs . Choleliths were located in the gallbladder in 20 dogs and in the bile ducts in 14 dogs . The most common abnormalities of the gallbladder, identified histologically, were chronic cholecystitis, mucosal hyperplasia, and pericholecystic inflammation . The most common abnormalities of the liver were cholestasis, hepatocellular degeneration, and periportal fibrosis . Survival rate of dogs that underwent cholecystectomy tended to be higher (86%) than that of dogs treated via cholecystotomy (50%) or cholecystectomy in combination with choledochotomy (33%) . Dogs that underwent medical treatment, abdominal exploratory, cholecystojenunostomy, choledochotomy, and sphincter of Oddiotomy died or were euthanatized because of redevelopment of clinical signs associated with cholelithiasis.(ABSTRACT TRUNCATED AT 250 WORDS) EMBO J, 1993 Apr, 12(4), 1265 - 75 Growth phase dependence of the activation of a bacterial gene for carotenoid synthesis by blue light; Fontes M et al.; Myxococcus xanthus responds to blue light by producing carotenoid pigments . A mutation at a gene named carC is known to block the metabolism of phytoene, a carotenoid precursor, and this gene has now been cloned and sequenced . We show here that gene carC, which is homologous to phytoene dehydrogenase genes from other organisms, is tightly regulated by light through a mechanism that operates only when the cells have reached the stationary phase or are starved of a carbon source . A genetic element that mediates the effect of the growth phase has been identified . Gene carC is integrated with another unlinked carotenogenic gene in a single 'light regulon' controlled by common trans-acting genetic elements . A potential -35 site for the binding of sigma factors has been found upstream of the carC transcriptional start . However, the -10 region shows no similarity with analogous sites at promoters of other Gram-negative bacteria. J Bacteriol, 1993 Apr, 175(7), 2157 - 61 Heat shock-induced axenic growth of Bdellovibrio bacteriovorus; Gordon RF et al.; The bdellovibrios are obligately predatory bacteria that attack other gram-negative bacteria . They grow only in the periplasmic space of prey unless they mutate to forms that can grow axenically . A culture medium that promoted enhanced growth of prey-independent bdellovibrios was developed . The ability of this medium to support the growth of prey-dependent bdellovibrios was tested under transcription-altering conditions . This approach tested the hypothesis that the inability to grow prey-dependent bdellovibrios in artificial media was rooted in both nutritional and transcriptional signal deficiencies . It was assumed that nutritional deficiencies had been resolved and that empirically applied artificial signals may evoke the expression of genes required for axenic growth of bdellovibrios . Prey-dependent bdellovibrios could be grown in PPYE medium (0.1% proteose peptone 3 and 0.03% Bacto yeast extract adjusted to pH 7.0 and supplemented with 3 mM MgCl2 and 2 mM CaCl2 after autoclaving) after heat shock, and subsequent rounds of growth occurred after additional heat shocks . Heat shock may have generated or simulated signals normally derived from prey. J Infect Dis, 1993 Apr, 167(4), 865 - 75 Human anti-endotoxin antibody HA-1A mediates complement-dependent binding of Escherichia coli J5 lipopolysaccharide to complement receptor type 1 of human erythrocytes and neutrophils;