Cytokine, 1994 Sep, 6(5), 530 - 6 Endotoxin and tumour necrosis factor do not cause mortality from caecal ligation and puncture; McMasters KM et al.; Macrophage tumour necrosis factor-alpha (TNF-alpha) production is thought to represent an important pathogenic mechanism by which Gram-negative sepsis is mediated . We compared the effects of caecal ligation and puncture (CLP) on endotoxin-sensitive (C3H/HeSnJ) and endotoxin-resistant (C3H/HeJ) mice . Mortality after CLP for C3H/HeSnJ mice compared with C3H/HeJ mice was not significantly different (32% and 55%, respectively) . When survivors were injected with lipopolysaccharide intraperitoneally on the 7th day after CLP, the mortality rate was 82% for C3H/HeSnJ mice versus 0% for C3H/HeJ mice (P < 0.0001) . Serum endotoxin levels at 24 h after CLP were only slightly elevated . Serum TNF levels and peritoneal macrophage TNF production were undetectable in C3H/HeJ mice and were only slightly elevated in C3H/HeSnJ mice by 24 h after CLP . Peritoneal macrophage mRNA levels for TNF-alpha, IL-1 beta, and I-A alpha displayed a similar pattern in the two strains of mice, with a 2- to 3-fold increase in TNF-alpha and IL-1 beta mRNA levels by 24 h and a sharp decrease in I-A alpha mRNA by 24 h . The cause of mortality in mice that undergo CLP cannot be attributed to overwhelming endotoxemia and/or TNF production. Shock, 1994 Sep, 2(3), 203 - 9 Cardiodynamic response to Escherichia coli endotoxemia: effects of fluid resuscitation; Zhong J et al.; We tested the influence of in vivo volume resuscitation on intrinsic contractile properties of left ventricular (LV) preparations of endotoxemic guinea pigs . Escherichia coli endotoxin (LPS)-injected animals were divided into nonresuscitated and resuscitated groups . Volume resuscitation improved cardiac output and stroke volume, increased arterial pH and body temperature, and decreased mortality . In isovolumetric LV preparations isolated 4 h after LPS injection, LV systolic pressures (in mmHg) preparations isolated 4 h after LPS injection, LV systolic pressures (in mmHg) of LPS with (42 +/- 3) and without (42 +/- 2) fluid resuscitation were consistently less than control values (70 +/- 3) . LV end-diastolic pressure-volume (compliance) decreased in LPS-nonresuscitated hearts, while LV compliance of LPS-resuscitated hearts was similar to control . Thus, intravascular volume expansion selectively improved LV diastolic compliance of LPS hearts without affecting LV systolic function . These findings suggest that LV systolic and diastolic dysfunctions associated with endotoxemia and Gram-negative sepsis may involve separate pathogenic mechanisms. Neurosurgery, 1994 Sep, 35(3), 484 - 90; discussion 491-2 Efficacy of prophylactic antibiotics for craniotomy: a meta-analysis; Barker FG 2nd; A meta-analysis of published randomized studies comparing prophylactic antibiotics to placebo in craniotomies was performed . Ten studies were examined; eight met criteria for inclusion into the meta-analysis . The analysis showed an advantage of antibiotics over placebo at the P < 10-8 level . Tests for homogeneity of effect size between the individual studies showed similar effects of antibiotic treatment between trials, despite variation in the randomization methods and antibiotic regimens used . No statistically significant difference was detected between antibiotic regimens that did or did not cover gram-negative organisms or between single- and multiple-dose regimens . Cumulative meta-analyses showed that this conclusion could have been confidently drawn by 1988, after only four of the eight eligible trials had been published . Trials published since that time have reinforced these conclusions but have not significantly altered them . Future studies should compare proposed new antibiotic regimens with one of those already demonstrated to be effective, not with a placebo. Lancet, 1994 Aug 13, 344(8920), 429 - 31 Relative concentrations of endotoxin-binding proteins in body fluids during infection; Opal SM et al.; Endotoxin initiates the systemic inflammatory response, haemodynamic changes, and multi-organ failure that may occur as a consequence of systemic gram-negative bacterial infection . The serum protein lipopolysaccharide-binding protein (LBP) binds to the lipid A component of bacterial endotoxin and facilitates its delivery to the CD14 antigen on the macrophage, where inflammatory cytokines are released and a cascade of host mediators is initiated . The neutrophil granular protein bactericidal/permeability-increasing protein (BPI) competes with LBP for endotoxin binding and functions as a molecular antagonist of LBP-endotoxin interactions . We have measured concentrations of both proteins in body fluids from 49 consecutive patients . In 16 of 17 samples of fluid from closed-space infections, BPI was present in greater concentration than LBP (median BPI/LBP ratio 7.6 {95% CI 2.32-22.1}) . The ratio of BPI and LBP was not significantly different from 1.0 in abdominal fluid from 10 patients with peritonitis (ratio 0.235 {0.18-0.47}), whereas the BPI/LBP ratio was low in 22 non-infected body fluids (0.01 {0.001-0.04}) and concentrations of both proteins approached those in normal human plasma . BPI concentrations were directly correlated with the quantity of neutrophils within clinical samples (rs = 0.81, p < 0.0001) . Thus, within abscess cavities BPI is available in sufficient quantities for effective competition with LBP for endotoxin . BPI may attenuate the local inflammatory response and the systemic toxicity of endotoxin release during gram-negative infections. Schweiz Rundsch Med Prax, 1994 Aug 9, 83(32), 865 - 9 {Non-surgical therapy of pancreatitis complications (pseudocyst, abscesses, stenoses)}; Singer MV et al.; Acute and chronic pseudocysts differ . Chronic pseudocysts develop during the evolution of chronic pancreatitis unrelated to a specific bout of clinically recognizable acute pancreatitis . Acute pseudocysts arise in conjunction with an episode of acute pancreatitis . Whereas until recently surgical therapy has been the standard treatment for acute (or chronic) pancreatic pseudocysts, a range of nonsurgical options has been developed . The most important nonsurgical treatment of all is to watch and wait . Pseudocysts following acute pancreatitis should be observed when they are truly asymptomatic and less than or equal to 6 cm in diameter and left alone if not increasing in size . Only if after a six-week observation period pancreatic pseudocysts increase in diameter and become symptomatic, percutaneous needle aspiration, catheter drainage or an endoscopic drainage procedure (cystogastrostomy/cystoduodenostomy) or ultimately operative drainage procedure should be considered . Antibiotic therapy should be considered for all patients presenting with pancreatic necrosis . They should be treated with drugs administered intravenously at the maximum recommended dose as early as possible after onset of symptoms, continued throughout at least the first two weeks of the disease . Moreover, they should be treated alone and/or in combination with antibiotics that are active against gram-negative organisms of intestinal origin, commonly isolated in necrotic tissue, pseudocysts and infected pancreatic abscesses, and that are capable of penetrating into the pancreatic juice and necrotic tissue (e.g . mezlocillin, cephalosporin, metronidazole) . Removal of pancreatic stones and pancreatic stenosis by endoscopic procedures in the treatment of pain in patients with chronic pancreatitis is still not an established and generally accepted treatment . Controlled trials to validate stenting and ESWL in chronic pancreatitis are needed. FEMS Microbiol Lett, 1994 Aug 1, 121(1), 11 - 8 Controlled expression of click beetle luciferase using a bacterial operator-repressor system; Enrique Vazquez M et al.; The bioluminescent phenotype conferred by luciferase genes in a particular bacterium has demonstrated to be one of the most versatile and useful methods to detect microorganisms . Genetic constructions derived from miniTn5 vectors have been constructed for the introduction and stable maintenance of the click beetle luciferase gene, lucOR, in various Gram-negative bacteria . To attenuate the expression in the environment where the marked strain has to survive (and to allow sensitive detection when desired) a DNA fragment containing the repressor gene lacIq and a Ptrc::lucOR fusion was cloned onto a suicide plasmid . This construction is able to express high luciferase levels only when induced by IPTG . Matings between Escherichia coli containing the suicide transposon vector and different recipient bacteria gave transposition frequencies from 10(-7) to 10(-5) . Strains with miniTn5-lucOR insertions showed luciferase activity induced by IPTG addition . The stringency of the regulation and the intensity of light emission depended on the tagged strain . This system allows stable maintenance of the marker and tight control of luciferase expression in the environment. Am J Physiol, 1994 Aug, 267(2 Pt 1), L137 - 44 Interleukin-8 is a major component of pleural liquid chemotactic activity in a rabbit model of endotoxin pleurisy; Boylan AM et al.; Gram-negative endotoxin induces production of the potent chemotactic factor interleukin-8 (IL-8) in vitro; however, the importance of IL-8 in endotoxin-induced inflammation in vivo is unknown . We asked whether IL-8 is an important contributor to chemotactic activity in acute inflammatory liquids formed in response to endotoxin, and, if present, what concentrations of IL-8 antigen are generated . For these studies, we cloned and expressed rabbit recombinant IL-8 (rrIL-8), developed specific anti-rabbit IL-8 monoclonal antibodies (mAb), and then used these reagents to develop assays to detect rabbit IL-8 bioactivity and measure rabbit IL-8 antigen . Escherichia coli endotoxin (20 ng/ml, n = 4, or 2,000 ng/ml, n = 4) was instilled into the pleural space of eight rabbits for 6 h . Rabbit IL-8 bioactivity in the endotoxin pleurisy samples was assayed by measuring the migration of rabbit neutrophils toward the pleural liquid under two different conditions: 1) after addition of an anti-IL-8 neutralizing mAb and 2) after desensitization of the neutrophils to rrIL-8 . Addition of the anti-IL-8 mAb decreased neutrophil migration toward the pleural liquid by 65 +/- 13 and 75 +/- 22% (mean +/- SE, after 20 and 2,000 ng/ml endotoxin, respectively; P < 0.01 compared with a control mAb) . Desensitization of neutrophils to rrIL-8 decreased their migration toward the pleural liquid by 72 +/- 5% (P = 0.03, compared with exposure of neutrophils to buffer alone.(ABSTRACT TRUNCATED AT 250 WORDS) South Med J, 1994 Aug, 87(8), 821 - 2 Pseudomonas vesicularis causing bacteremia in a child with sickle cell anemia; Oberhelman RA et al.; Pseudomonas vesicularis is a gram-negative rod infrequently found in environmental sources or clinical specimens . We report a case of bacteremia due to Pseudomonas vesicularis in a child with sickle cell anemia, fever, and pneumonia . This is the first published report of invasive P vesicularis in a child. Crit Care Med, 1994 Aug, 22(8), 1227 - 34 An evaluation of the hemodynamic effects of HA-1A human monoclonal antibody; Kett DH et al.; OBJECTIVES: We sought to determine whether there might be acute changes in hemodynamics attributable to HA-1A, a monoclonal antibody to endotoxin, in patients with presumed Gram-negative sepsis . DESIGN: Post hoc analysis of a multicenter, randomized, double-blind, placebo-controlled study . PATIENTS: A total of 543 patients with severe sepsis presumed to be caused by Gram-negative bacteria who were enrolled in a clinical trial to evaluate the efficacy and safety of HA-1A human monoclonal antibody . INTERVENTIONS: Patients were randomly assigned to receive either 100 mg of HA-1A or placebo . MEASUREMENT AND MAIN RESULTS: Patients were grouped by the study drug, HA-1A, or placebo, and the presence or absence of Gram-negative bacteremia . Hemodynamic variables were monitored from before, until 72 hrs after infusion of the study drug . For the entire study population (n = 543), no changes over time attributable to study drug were noted in the mean arterial pressure (p > .19), heart rate (p > .53) or the need for vasopressor administration (p > .62) . One hundred ninety-seven patients underwent pulmonary artery catheterization and had hemodynamic data available from before the infusion of HA-1A or placebo until at least 12 hrs after infusion . Evaluating all 197 patients on an intent to treat basis demonstrated no significant differences over time in cardiac index (p > .15), oxygen delivery index (p > .43), or left ventricular stroke work index (p > .48) between patients who received HA-1A and those patients receiving placebo . Grouping patients by the presence of Gram-negative bacteremia and study drug received also failed to demonstrate any significant difference attributable to HA-1A in mean arterial pressure (p > .54), heart rate (p > .84), cardiac index (p > .13), oxygen delivery index (p > .05), or left ventricular stroke work index (p > .48) between populations . CONCLUSION: There is no apparent relationship between the administration of HA-1A, the presence of Gram-negative bacteremia, and hemodynamic profiles over the 72-hr study period. Clin Chem, 1994 Aug, 40(8), 1575 - 9 Endotoxin binding to platelets in blood from patients with a sepsis syndrome; Salden HJ et al.; Endotoxin, the lipopolysaccharide cell wall constituent of Gram-negative bacteria, produces symptoms of the Gram-negative sepsis syndrome . By measuring endotoxin in blood from septic patients it may be possible to select a subpopulation of patients in which mortality can be prevented by treatment with anti-endotoxin antibodies . We evaluated the performance of an endotoxin-free blood-collection tube . Within-run and between-run CVs of our endotoxin assay were 4-18% and 8-20%, respectively . In endotoxin-positive samples (LPS > or = 6 ng/L), the concentration of endotoxin in platelet-rich plasma was significantly higher (P < 0.001) than in platelet-poor plasma . Apparent binding of endotoxin to platelets ranged from 0% to 92% . The correlation between the apparent percentage binding of LPS to platelets and the platelet count in platelet-rich plasma is linear and positive, but LPS is not bound solely to platelets . We conclude that endotoxin must be measured in platelet-rich plasma. Acta Paediatr, 1994 Aug, 83(8), 797 - 801 Eosinophilia in newborn infants; Patel L et al.; To evaluate the clinical significance of eosinophilia in newborn infants, 261 admissions to the neonatal unit over a 12-month period were studied retrospectively; 33 babies with eosinophilia (> 1.0 x 10(9)/l) were studied . Clinical and laboratory data for the first month of life were compared, where available, between gestational age-matched pairs with and without eosinophilia . Of the 33 babies with eosinophilia, 23 were > 26 weeks' gestation and all had age-matched controls; 10 were < or = 26 weeks' gestation but had no appropriate gestational age-matched controls . Babies > 26 weeks' gestation with eosinophilia had a significantly higher number of septic episodes than controls: 20 of 23 versus 4 of 23 . All 10 babies < or = 26 weeks' gestation with eosinophilia developed sepsis . Infections with gram-negative organisms and necrotizing enterocolitis occurred only in babies who developed eosinophilia . In 5 babies no cause for the eosinophilia was found . In conclusion, eosinophilia in the newborn is usually explainable and is most often associated with infection. Microbiology, 1994 Aug, 140 ( Pt 8), 2143 - 51 Nucleotide sequence and characterization of the Rhodobacter sphaeroides glnB and glnA genes; Zinchenko V et al.; The glnA gene of Rhodobacter sphaeroides encoding glutamine synthetase (GS) has been cloned and sequenced . Molecular analysis revealed that there is a glnB gene upstream of glnA, in a single glnBA operon . A putative glnAp1-type promoter sequence, a consensus ntrC gene product binding site and a consensus upstream activator sequence were detected upstream of the glnB gene . The deduced amino acid sequences of the GS and GlnB proteins of R . sphaeroides showed strong homology with the same proteins from other Gram-negative bacteria . The sequence of the glnA gene isolated from glutamine auxotroph Gln83 was also determined . The glnA83 mutation was shown to result in premature termination of GS synthesis and formation of a 17 kDa C-truncated GS which could be complemented by a 5'-truncated glnA gene which encodes a 30 kDa N-truncated GS . This phenomenon is characteristic for interallelic complementation. East Afr Med J, 1994 Aug, 71(8), 501 - 2 Highly resistant bacterial osteomyelitis in Somalia; Jumale AH et al.; Chronic osteomyelitis is a major health problem in the aftermath of the conflict in Somalia . We studied the microbiology of chronic osteomyelitis among 30 patients in a large hospital in Mogadishu . Gram-negative bacteria were isolated from 77% of patients; most of these isolates were highly resistant to standard antibiotic agents but all were sensitive to ciprofloxacin, an oral fluoroquinolone antibiotic . Ciprofloxacin may prove useful in the management of chronic osteomyelitis due to highly resistant gram-negative bacteria. Immunopharmacol Immunotoxicol, 1994 Aug, 16(3), 449 - 61 Downregulation of human polymorphonuclear cell activities exerted by microorganisms belonging to the alpha-2 subgroup of Proteobacteria (Afipia felis and Rochalimaea henselae); Fumarola D et al.; Intracellular pathogens have evolved effective mechanisms in order to survive in an intracellular environment, thus avoiding destruction by phagocytic cells . In this regard, a correlation between resistance to phagocytic killing and expression of pathogenic potency has been established . In this report, we have studied the interaction between human polymorphonuclear cells (PMN) and two gram-negative microorganisms, Afipia felis and Rochalimaea henselae, which belong to the alpha-2 subgroup of the class Proteobacteria . A . falis has been previously proposed as the causative agent of Cat Scratch Disease (CSD), but several recent lines of evidence attribute a major role to R . henselae . Of note, CSD is a syndrome characterized by a chronic lymphoadenopathy, involving macrophages and endothelial cells with a progression towards a granulomatous process and/or angiogenesis . Since members of the alpha-2 subgroup of Proteobacteria have the property to survive intracellularly, we have evaluated the effects exerted by A . felis and R . henselae on human PMN in terms of chemotaxis locomotion, degranulation and oxidative metabolism . Results will show an impairment of PMN activities as a consequence of the challenge with both microrganisms . In particular, inhibition of PMN oxidative function occurred either as result of a direct exposure to both A . felis and R . henselae or when PMN were primed by bacteria for the N-formyl-methionyl-leucyl-phenylalanine enhancement of the oxidative burst . These findings may account for the ability of A . felis and R . henselae to survive within PMN as expression of a further mechanism of pathogenic potency, influencing also the nature and the evolution of inflammatory response in the lesion sites. FEBS Lett, 1994 Jul 25, 349(1), 69 - 74 Characterisation of the porin of Rhodobacter capsulatus 37b4 in planar lipid bilayers; Bishop ND et al.; The outer membrane of Gram-negative bacteria contains aqueous channels, porins, which aid the diffusion of small hydrophilic molecules across it . Escherichia coli, as enteric bacteria, are able to survive a hostile environment of proteases, surfactants, and drastic changes of osmotic pressure . Rhodobacter capsulatus is not an enteric bacterium and as such has not evolved to resist the same challenges . Porins, which have molecular weight of approximately 35 kDa, form trimeric channels with a solute exclusion limit of about 600 Da . Most of them open and close in a controlled manner as a function of p.d . This function is little understood at present . The functional properties of single trimers of the major porin of Rhodobacter capsulatus 37b4 have been investigated in planar artificial bilayers . On application of a suitable p.d . the observed trimer closes in approximately three equal steps . The behaviour is completely symmetrical as regards closure in response to p.d.'s of opposite polarity and is strongly cation selective. Ann Intern Med, 1994 Jul 1, 121(1), 1 - 5 Treatment of septic shock with human monoclonal antibody HA-1A . A randomized, double-blind, placebo-controlled trial . CHESS Trial Study Group; McCloskey RV et al.; OBJECTIVE: To compare the effectiveness of 100 mg of HA-1A and placebo in reducing the 14-day all-cause mortality rate in patients with septic shock and gram-negative bacteremia in the Centocor: HA-1A Efficacy in Septic Shock (CHESS) trial, and to assess the safety of 100 mg of HA-1A given to patients with septic shock who did not have gram-negative bacteremia . DESIGN: Large, simple, group-sequential, randomized, double-blind, multicenter, placebo-controlled trial . SETTING: 603 investigators at 513 community and university-affiliated hospitals in the United States . PATIENTS: Within 6 hours before enrollment, the patients had been in shock with a systolic blood pressure of less than 90 mm Hg after adequate fluid challenge or had received vasopressors to maintain blood pressure . These episodes of shock began within 24 hours of enrollment . A presumptive clinical diagnosis of gram-negative infection as the cause of the shock episode and a commitment from the patients' physicians to provide full supportive care were required . MEASUREMENTS: Blood cultures were obtained within 48 hours of enrollment, and death at day 14 after treatment was recorded . Adverse events occurring within 14 days after enrollment were also tabulated . RESULTS: 2199 patients were enrolled; 621 (28.2%) met all enrollment criteria, received HA-1A or placebo, and had confirmed gram-negative bacteremia . Mortality rates in this group were as follows: placebo, 32% (95 and HA-1A, 33% (109 of 328) (P = 0.864, Fisher exact test, two-tailed; 95% CI for the difference, -6.2% to 8.6%) . Mortality rates in the patients without gram-negative bacteremia were as follows: placebo, 37% (292 of 793) and HA-1A, 41% (318 of 785) (P = 0.073, Fisher exact test, one-tailed; CI, -0.8% to 8.8%) . CONCLUSIONS: In this trial, HA-1A was not effective in reducing the 14-day mortality rate in patients with gram-negative bacteremia and septic shock . These data do not support using septic shock as an indication for HA-1A treatment . If HA-1A is effective in reducing the mortality rate in patients dying from endotoxemia, these patients must be identified using other treatment criteria. Can J Microbiol, 1994 Jul, 40(7), 555 - 60 Description of bacteria able to degrade isoquinoline in pure culture; Aislabie J et al.; Isoquinoline is a nitrogen heterocyclic compound that is associated with coal- and oil-derived wastes . Four strains of bacteria able to degrade isoquinoline in pure culture were isolated from sites known to be contaminated with oil . Isoquinoline was used as the sole source of carbon and nitrogen by these isolates . Isoquinoline was initially transformed to 1-hydroxyisoquinoline, which accumulated in the broth culture, and then disappeared . The four strains isolated were Gram negative, aerobic, rod-shaped bacteria with polar flagella . The strains have been presumptively identified as members of the family Comamonadaceae. Clin Exp Immunol, 1994 Jul, 97 Suppl 1, 69 - 72 Polyclonal intravenous immune globulin for prevention and treatment of infections in critically ill patients; Cometta A et al.; Infections remain the leading cause of death among patients admitted to intensive care units (ICU) . Infections due to Gram-negative bacteria are both frequent and difficult to treat . The poor outcome of such infections has been attributed to the endotoxin . The high mortality rate related to Gram-negative sepsis has prompted the testing of new, adjunctive therapies to prevent and treat infections in critically ill patients . Immunotherapy or immunoprophylaxis have long been investigated in this context . Passive immunotherapy consists of the administration of immune plasma or serum, or standard or hyperimmune purified immune globulins . Several clinical studies using such preparations to treat critically ill patients are reviewed in this article . While two studies using hyperimmune plasma or serum appeared to be successful, two studies using hyperimmune globulin failed to show a beneficial effect in the treatment or the prevention of Gram-negative septic shock . Regarding the infusion of standard intravenous immune globulin (IVIG) two studies have demonstrated a substantial benefit in the prevention of severe infections; the reduction of nosocomial pneumonia recorded in both trials and the shortness of stay in ICU may also afford savings in hospital costs . The cost effectiveness of such prophylactic administration of IVIG is worthy of further investigation. Crit Care Med, 1994 Jul, 22(7), S12 - 8 Current prospects for the treatment of clinical sepsis; Suffredini AF; OBJECTIVES: To review the role of antimediator therapy in the inflammatory cascade associated with sepsis, and to review the status of animal and clinical studies being conducted on novel therapies for septic shock . DATA SOURCES: Information presented at the 22nd Educational and Scientific Meeting of the Society of Critical Care Medicine on June 9-13, 1993 in New York City was reviewed, along with supportive documentation from the English language literature . STUDY SELECTION: Controlled animal studies that provide evidence for the effectiveness of antiendotoxin and anticytokine therapies . The preliminary results of selected clinical trials are also presented . DATA EXTRACTION: This review focuses on data describing the potential role of mediator antagonists in the treatment of septic shock . DATA SYNTHESIS: Information concerning the effectiveness and tolerability of these therapies has been integrated into a description of emerging therapies for septic shock . CONCLUSIONS: Clinical trials of antiendotoxin antibodies have not shown them to have therapeutic benefit . New agents that neutralize or antagonize the cellular effects of endotoxin may provide an alternative means to inhibit endotoxin effects during severe Gram-negative infections . Anti-interleukin-1 and antitumor necrosis factor-alpha therapies have demonstrated efficacy in animal models, but the results have been inconsistent in human trials . Preliminary results from clinical trials of cytokine antagonists suggest that these therapies may be effective in the most severely ill patients . Further clinical trials will be required to determine the therapeutic role of these agents in septic shock. Am J Respir Crit Care Med, 1994 Jul, 150(1), 179 - 83 Increased salivary exoglycosidase activity during critical illness; Quinn MO et al.; Exoglycosidases remove peripheral monosaccharides from oligosaccharides and hence are capable of altering respiratory epithelial cell surface carbohydrates . We obtained saliva and tracheal secretions from 34 critically ill patients and saliva from 23 healthy subjects . Compared with the normal subjects, the ill patients had large amounts of mannosidase, fucosidase, hexosaminidase, and sialidase activity . Sialidase increased adherence of several gram-negative bacteria to epithelial cell monolayers and pure glycoproteins . Pretreatment of glycoproteins with some of the patients' saliva samples also increased bacterial adherence to the glycoproteins . We conclude that respiratory tract exoglycosidase activity increases during critical illness . By altering normal cell surface carbohydrates, exoglycosidases may facilitate bacterial adherence and respiratory tract colonization. Am J Respir Crit Care Med, 1994 Jul, 150(1), 131 - 4 Buccal cell carbohydrates are altered during critical illness; Weinmeister KD et al.; Cell-surface carbohydrates mediate the adherence of many pathogenic bacteria to epithelial cells . Because gram-negative bacteria adhere especially well to respiratory epithelial cells obtained from severely ill patients, we compared respiratory epithelial cell-surface carbohydrate levels of normal subjects with those of critically ill patients . Lectins were used to quantitate the amount of mannose, galactose, fucose, and sialic acid on buccal and tracheal cells . Fifteen critically ill patients, 20 normal subjects, and 10 minimally ill hospitalized patients were studied . The severely ill patients' buccal cells had decreased amounts of sialic acid and galactose . No differences were found between the normal and critically ill patients' tracheal-cell carbohydrates . The results obtained with a sialic acid-specific lectin were confirmed by direct measurement of buccal-cell sialic acid . We conclude that severely ill patients have decreased amounts of galactose and sialic acid on their upper-airway epithelial cells, and that loss of these two monosaccharides may explain the high prevalence of gram-negative bacterial colonization and pneumonia in the critically ill. J Bacteriol, 1994 Jul, 176(13), 4173 - 6 Transformation of Actinomyces spp . by a gram-negative broad-host-range plasmid; Yeung MK et al.; The gram-negative broad-host-range vector pJRD215 was transferred by electroporation into strains of Actinomyces viscosus or Actinomyces naeslundii at efficiencies which ranged from 10(2) to 10(7) transformants per microgram of plasmid DNA . The Actinomyces transformants expressed pJRD215-encoded resistance to kanamycin and streptomycin . Moreover, the transforming plasmid DNA had not undergone any deletions or rearrangements, nor had it integrated into the genomes of these strains. J Bacteriol, 1994 Jul, 176(13), 3825 - 31 A family of extracytoplasmic proteins that allow transport of large molecules across the outer membranes of gram-negative bacteria; Dinh T et al.; Seventeen fully sequenced and two partially sequenced extracytoplasmic proteins of purple, gram-negative bacteria constitute a homologous family termed the putative membrane fusion protein (MFP) family . Each such protein apparently functions in conjunction with a cytoplasmic membrane transporter of the ATP-binding cassette family, major facilitator superfamily, or heavy metal resistance/nodulation/cell division family to facilitate transport of proteins, peptides, drugs, or carbohydrates across the two membranes of the gram-negative bacterial cell envelope . Evidence suggests that at least some of these transport systems also function in conjunction with a distinct outer membrane protein . We report here that the phylogenies of these proteins correlate with the types of transport systems with which they function as well as with the natures of the substrates transported . Characterization of the MFPs with respect to secondary structure, average hydropathy, and average similarity provides circumstantial evidence as to how they may allow localized fusion of the two gram-negative bacterial cell membranes . The membrane fusion protein of simian virus 5 is shown to exhibit significant sequence similarity to representative bacterial MFPs. Blood, 1994 Jul 1, 84(1), 27 - 35 Mice lacking both macrophage- and granulocyte-macrophage colony-stimulating factor have macrophages and coexistent osteopetrosis and severe lung disease; Lieschke GJ et al.; Mice deficient in granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF, CSF-1) were generated by interbreeding GM-CSF-deficient mice generated by gene targeting (genotype GM-/-) with M-CSF-deficient osteopetrotic mice (genotype M-/-, op/op) . Mice deficient in both GM-CSF and M-CSF (genotype GM-/-M-/-) are viable and have coexistent features corresponding to mice deficient in either factor alone . Like M-CSF-deficient mice, they have osteopetrosis and are toothless because of failure of incisor eruption . Like GM-CSF-deficient mice, they have a characteristic alveolar-proteinosis-like lung pathology, but it is more severe than that of GM-CSF-deficient mice and is often fatal . In particular, in GM-/-M-/- mice the accumulation of lipo-proteinaceous alveolar material is more marked, and bacterial pneumonic infections are more prevalent and more extensive, particularly involving Gram-negative bacteria . Neutrophilia consistently accompanies pulmonary infections, and some older GM-/-M-/- mice have polycythemia . Survival of GM-/-M-/- mice is significantly reduced compared with mice deficient in either factor alone, and all GM-/-M-/- mice have broncho- or lobar-pneumonia at death . These observations indicate that in vivo, M-CSF is involved in modulating the consequences of GM-CSF deficiency in the lung . Interestingly, GM-/-M-/- mice have circulating monocytes at levels comparable with those in M-CSF-deficient mice and the diseased lungs of all GM-/-M-/- mice contain numerous phagocytically active macrophages, indicating that in addition to GM-CSF and M-CSF, other factors can be used for macrophage production and function in vivo. Infect Immun, 1994 Jul, 62(7), 2800 - 5 Affinity of the C9 molecule for the C5b-8 complex compared with that for the complex containing C9 molecules; MacKay SL et al.; Gram-negative bacterial cells exposed to a complement source may carry membrane attack complexes containing variable numbers of C9 molecules per C5b-8 site . In order to investigate the assembly of this complex, the ability of C9 molecules to bind to C5b-8 complexes was compared with the binding characteristics of C9 for C5b-8 complexes containing variable numbers of bound C9 molecules . The apparent dissociation constant (Kd) of the C9 molecule for the C5b-8 site on a complement-sensitive strain of Escherichia coli was 1.2 (+/- 0.15) nM at 0 degree C . These conditions allow the binding of one C9 molecule per C5b-8 site . The C5b-8 site containing one C9 molecule bound a second C9 molecule at 0 degree C only after incubation at 37 degrees C . The binding of C9 to a C5b-8 site containing one C9 molecule was found to be 1.3 (+/- 0.2) nM . Therefore, the presence of a C9 molecule did not significantly alter the binding capacity of the C5b-8 site for additional C9 molecules . A similar result was obtained by using rabbit erythrocytes bearing either C5b-8 sites or C5b-8 sites containing one molecule of C9 per complex at 0 degree C . The similarity of binding characteristics for the first and second C9 molecules argues that the initial C9 molecule in the complex does not affect the binding of subsequent C9 molecules . This suggests that a unique C9 binding site that does not involve previously bound C9 molecules may exist on the forming membrane attack complex. Clin Microbiol Rev, 1994 Jul, 7(3), 277 - 89 Molecular adjuvants and immunomodulators: new approaches to immunization; Johnson AG; Epitopes on microbial antigens responsible for protective immunity have begun to be identified and isolated, and their chemical structures have been determined . Ensuing knowledge of their weak immunizing capacity per se has led to an appreciation of the need for adjuvants to increase the immunogenicity of these low-molecular-weight synthetic structures . As such, a recent surge in adjuvant research has emerged . Accordingly, this review will highlight a number of those adjuvant substances whose activity in animals indicates a potential use in human vaccines . In addition, the potential of several well-defined substances, termed immunomodulators, which nonspecifically stimulate resistance of animals to multiple 50% lethal doses of microbial challenge is described . Among the most extensively characterized adjuvants of microbial origin discussed in detail are (i) the lipopolysaccharides isolated from gram-negative bacteria and their nontoxic analogs, (ii) the synthetic muramyl dipeptides and their multiple analogs, and (iii) the synthetic polyribonucleotide complexes, mimicking the interferon-inducing capacity of viruses . Discussed also are the heat-labile enterotoxin of Escherichia coli, the nonionic block copolymers, the saponins, a quinolamine derivative, and the hormone dihydroepiandrosterone. Circ Shock, 1994 Jul, 43(3), 115 - 21 Renal interleukin-1 expression during endotoxemia and gram-negative septicemia in conscious rats; Laszik Z et al.; Endotoxin-induced cytokines such as interleukin-1 (IL-1) and tumor necrosis factor (TNF) are thought to contribute to the proinflammatory effects of endotoxin in gram-negative infections . Using a conscious rat model of sepsis, induced by intravenous challenge with LD95 doses of endotoxin (n = 24) or live Escherichia coli (E . coli) (n = 24), we examined frozen sections of kidney at various intervals for evidence of IL-1 alpha and TNF alpha expression . A transient glomerular endothelial IL-1 alpha expression was demonstrated at 30 and 90 min after initiation of the sepsis in both endotoxin and E . coli-treated animals using immunohistochemistry . The endothelial IL-1 alpha expression as determined by immunohistochemistry occurred at the same time as IL-1 alpha mRNA expression, as determined by Northern blot analysis . The glomerular endothelial IL-1 alpha expression coincided with a slight but significant increase in the number of the glomerular polymorphonuclear leukocytes as identified by naphthol AS-D chloroacetate esterase enzyme histochemical reaction . Glomerular endothelial IL-1 alpha expression was virtually absent by 180 and 360 min . No TNF alpha expression was detected in the renal tissues at any time interval . Neither alpha-naphthyl acetate esterase-positive nor acid phosphatase-positive monocytes/macrophages were identified in the glomeruli . Our findings provide direct in vivo evidence that the IL-1 alpha gene product is expressed locally in the kidney by glomerular endothelial cells in this septic rat model. Circ Shock, 1994 Jul, 43(3), 107 - 14 Monophosphoryl lipid A protects against endotoxic shock via inhibiting neutrophil infiltration and preventing disseminated intravascular coagulation; Yao Z et al.; The major objective of the present study was to determine the effects of a partial structure of the lipid A moiety of gram-negative lipopolysaccharide, monophosphoryl lipid A (MLA), on endotoxin-induced mortality and disseminated intravascular coagulation (DIC) in rats . A second objective was to examine the role of polymorphonuclear neutrophil invasion to visceral organs, including lung, liver, heart, and kidney in the pathogenesis of the compromised multiorgan function which occurs in endotoxic shock . Finally, a third aim was to determine if the potential protective effects of MLA might be mediated via inhibiting neutrophil invasion to various visceral organs . Male Sprague-Dawley rats (220-260 g) were fasted over night and used the following day . In control rats, endotoxin (S . abortus equi LPS, 15 mg/kg, i.v.) produced a 89% mortality at 48 hr following its administration, and gross pathological and laboratory signs of DIC at 3 hr after injection . The latter included increased serum fibrin(ogen) degradation products (FDP, 24.00 +/- 7.81 vs . 0 micrograms/ml, P < .05), prothrombin time (PT, 16.20 +/- 1.12 vs . 13.03 +/- 0.20 sec, P < .05), and activated partial thromboplastin time (APTT, 32.70 +/- 3.83 vs . 20.11 +/- 0.60 sec, P < .05), and decreased plasma fibrinogen (233.2 +/- 41.6 vs . 406.3 +/- 23.2 mg/dl, P < .05) as well as evidence of gross visceral hemorrhage . Pretreatment with MLA (5 mg/kg) for 24 hr produced a marked reduction in endotoxin-induced mortality at 48 hr (0% versus 89% in controls) and inhibited all of the manifestations of DIC produced by endotoxin.(ABSTRACT TRUNCATED AT 250 WORDS) PDA J Pharm Sci Technol, 1994 Jul-Aug, 48(4), 197 - 204 Validation of dry heat inactivation of bacterial endotoxins; Hecker W et al.; Bacterial endotoxins (ETs) are lipopolysaccharides from the cell wall of Gram negative bacteria . ETs get into the environment as a result of autolytic desintegration of the bacterial cells . There exist a number of depyrogenation methods, either serving to remove or to inactivate ET . The most common means of ET inactivation is dry heat . Unfortunately no uniform regulation exists describing the conditions for sufficient ET inactivation . While the USP and FDA require an ET reduction of 3 log steps, no European regulation exists regarding depyrogenation of final containers for parenterals . However, the Ph . Eur . specifies the temperature and time conditions for depyrogenation of glassware in the pyrogen test monograph, allowing to choose between the two variants 250 degrees C/30 minutes or 200 degrees C/60 minutes which are not equivalent . In the present study those conditions for depyrogenation of glass containers in production of parenterals were investigated which, on the one hand, are technically feasible and, on the other hand, comply with the requirements of the main Pharmacopeias; furthermore, an ET preparation suitable for validation studies was selected . The preparation of ET indicators, the dry-heat inactivation and the recovery of ET are described in detail . Based on the results obtained, it is recommended to follow a defined treating temperature and period for safe depyrogenation of glass containers for parenterals, which results in a 4 log step reduction in ET without fillers . Thereby the USP/FDA requirement for a 3 log step reduction as well as the 200 degrees C/60 minutes requirement variant given in the Ph . Eur . can be fulfilled. Int J Syst Bacteriol, 1994 Jul, 44(3), 379 - 86 The phloem-limited bacterium of greening disease of citrus is a member of the alpha subdivision of the Proteobacteria; Jagoueix S et al.; Using the PCR, we amplified the 16S ribosomal DNAs (rDNAs) of an Asian strain and an African strain of the uncultured, gram-negative, walled, phloem-limited bacterium-like organism (BLO) associated with citrus greening disease . We evaded coamplification of chloroplast 16S rDNA by using restriction enzymes; the chloroplast 16S rDNA was sensitive to BclI digestion and resistant to EcoRI digestion, while the 16S rDNA of the BLO was resistant to BclI digestion and sensitive to EcoRI digestion . The 16S rDNA of the African BLO strain was amplified from BclI-digested DNA extracted from infected periwinkle leaf midribs . The Asian strain was isolated from plant extract by using a specific monoclonal antibody coated onto the surface of a PCR tube . The 16S rDNAs of the two BLO strains were cloned and sequenced . Comparisons with sequences of 16S rDNAs obtained from the GenBank data base revealed that the two citrus greening disease BLOs belong to the alpha subdivision of the class Proteobacteria . Even though their closest relatives are members of the alpha-2 subgroup, these BLOs are distinct from this subgroup as we observed only 87.5% homology between the 16S rDNAs examined . Therefore, the two BLOs which we studied probably are members of a new lineage in the alpha subdivision of the Proteobacteria . We propose the trivial name "liberobacter" for this new group of bacteria and will wait until additional characteristics have been determined before we propose a formal name. J Immunol, 1994 Jul 1, 153(1), 287 - 99 Discordant adaptation of human peritoneal macrophages to stimulation by lipopolysaccharide and the synthetic lipid A analogue SDZ MRL 953 . Down-regulation of TNF-alpha and IL-6 is paralleled by an up-regulation of IL-1 beta and granulocyte colony-stimulating factor expression; Knopf HP et al.; Human peritoneal macrophages were exposed to increasing doses of LPS or a synthetic lipid A analogue (SDZ MRL 953) and production of the cytokines IL-1 beta, IL-6, TNF-alpha, and G-CSF was assessed at the protein and mRNA level . Cells were also prestimulated with low doses of LPS and SDZ MRL 953 to study their adaptation to a secondary challenge with high doses of LPS . The ability of macrophages to produce high levels of TNF-alpha and IL-6 after stimulation with LPS could be relieved almost completely by preincubating cells with low doses of LPS . Decreases of TNF-alpha and IL-6 production resulted from inhibition of gene transcription and/or changes in mRNA stability, as transcript levels of these cytokines were down-modulated by the process of LPS adaptation . Surprisingly, however, adapted cells were able to synthesize even larger quantities of G-CSF and IL-1 beta when exposed to a secondary LPS challenge . mRNA levels of the adapted cells remained unaltered for IL-1 beta, but were slightly increased for G-CSF as assessed by Northern blot analysis . High doses of the synthetic lipid A analogue SDZ MRL 953 were also able to adapt macrophages to a secondary LPS challenge by down-regulating TNF-alpha and IL-6 production, whereas priming secretion of G-CSF and IL-1 beta as well . We describe here the discordant adaptation of human peritoneal macrophages to a secondary LPS stimulus in vitro . These findings appear to have ramifications for the in vivo endotoxin response during inflammation and also Gram-negative septicemia. Proc Natl Acad Sci U S A, 1994 Jun 21, 91(13), 6017 - 20 Protection of mice from endotoxic death by 2-methylthio-ATP; Proctor RA et al.; The lethal effects of endotoxin, a bacterial product shed into the blood during bacteremia, are thought to be due to macrophage release of mediators such as tumor necrosis factor alpha and interleukin 1 . Although much is known about the pathophysiology of endotoxemia, relatively little is known about the cellular signaling mechanisms that are involved . The data in this study suggest that extracellular adenine nucleotides can influence the development of endotoxin shock . An adenine nucleotide analog, 2-methylthio-ATP, inhibited the endotoxin-stimulated release of toxic mediators (i.e., tumor necrosis factor alpha and interleukin 1), and it protected mice from endotoxin-induced death . These studies suggest a fundamental and unusual role for adenine nucleotides on endotoxin action, and they provide a potentially new therapeutic approach for the control of the pathophysiology of Gram-negative septicemia. Arch Intern Med, 1994 Jun 13, 154(11), 1241 - 9 Projected impact of monoclonal anti-endotoxin antibody therapy; Bates DW et al.; BACKGROUND: The goals of this study were to evaluate the criteria for administration of HA-1A monoclonal antibody therapy from the HA-1A trial in patients with suspected gram-negative bacteremia and to evaluate the accuracy with which Bone's criteria for sepsis syndrome identify patients with gram-negative bacteremia . METHODS: This prospective cohort study included 1509 episodes in which hospitalized patients had blood cultures performed in an urban tertiary-care hospital . The main outcome measures were gram-negative bacteremia and gram-negative sepsis . RESULTS: Of 1509 episodes, 115 (8%) represented bacteremia and 40 (3%) included gram-negative rods . Of these 40 patients, nine died in the hospital, including five patients who had gram-negative sepsis; all five had another rapidly fatal disease . Using criteria for treatment and exclusions from the HA-1A trial, three of the patients with gram-negative bacteremia would have been treated, while at least 52 patients without gram-negative bacteremia might have received HA-1A therapy (positive predictive value of criteria, 5.5%) . Of the 1509 episodes, sepsis syndrome as defined by Bone was present in 34 (2.3%) . While 32 of the 34 patients had suspected gram-negative bacteremia, only five had blood cultures positive for gram-negative bacteria . CONCLUSIONS: In this population, current criteria for administration of monoclonal anti-endotoxin antibody therapy were not sensitive or specific for gram-negative bacteremia, and many patients with gram-negative sepsis were too ill from other conditions to benefit . Indiscriminate use of these therapies could thus be costly yet yield few benefits . To identify patients who should receive novel therapies, better risk-stratification methods and cost-effectiveness analysis are needed. Metabolism, 1994 Jun, 43(6), 691 - 6 Regulatory factors in the development of fatty infiltration of the liver during gram-negative sepsis; Lanza-Jacoby S et al.; To further understand the development of fatty liver during gram-negative sepsis, we measured fatty acid uptake in addition to esterification and secretion of lipids by freshly isolated hepatocytes from fasted and fed control and Escherichia coli-treated rats . Rats were made septic by intravenous (IV) injection of 8 x 10(7) live E coli colonies per 100 g body weight . For the fasted groups, food was removed after E coli injection . Fed rats received a nutritionally adequate diet intragastrically for 5 days before and 24 hours after inducing sepsis . Twenty-four hours after E coli injection, the esterification of newly synthesized fatty acids, as measured by 3H2O incorporation, and the esterification of exogenous fatty acids, measured from 14C-palmitate incorporation, into triglyceride (TG), total cholesterol, and total phospholipid phosphorus were significantly greater in hepatocytes from fasted septic rats compared with their control rats . In fed septic rats, esterification of 14C-palmitate into TG was fourfold greater than in the fed control rats . The increased rates of esterification in hepatocytes from fasted and fed septic rats were not accompanied by an increase in the labeled TG in the medium . This inability to secrete the additional TG that the hepatocytes produce resulted in a higher concentration of cellular TG in fasted and fed septic rats than in their controls . The enzymes glycerol-3-phosphate acyltransferase (GPAT) and phosphatidate phosphohydrolase (PPH) do not appear to be factors contributing to the increased TG synthesis, since the increase in enzyme activity was not accompanied by a similar increase in TG synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) J Clin Invest, 1994 Jun, 93(6), 2600 - 7 Prenatal immune challenge alters the hypothalamic-pituitary-adrenocortical axis in adult rats; Reul JM et al.; We investigated whether non-abortive maternal infections would compromise fetal brain development and alter hypothalamic-pituitary-adrenocortical (HPA) axis functioning when adult . To study putative teratogenic effects of a T cell-mediated immune response versus an endotoxic challenge, 10-d-pregnant rats received a single intraperitoneal injection of 5 x 10(8) human red blood cells (HRBC) or gram-negative bacterial endotoxin (Escherichia coli LPS: 30 micrograms/kg) . The adult male progeny (3 mo old) of both experimental groups showed increased basal plasma corticosterone levels . In addition, after novelty stress the HRBC group, but not the LPS group, showed increased ACTH and corticosterone levels . Both groups showed substantial decreases in mineralocorticoid (MR) and glucocorticoid receptor (GR) levels in the hippocampus, a limbic brain structure critical for HPA axis regulation, whereas GR concentrations in the hypothalamus were unchanged and in anterior pituitary were slightly increased . HRBC and LPS indeed stimulated the maternal immune system as revealed by specific anti-HRBC antibody production and enhanced IL-1 beta mRNA expression in splenocytes, respectively . This study demonstrates that a T cell-mediated immune response as well as an endotoxic challenge during pregnancy can induce anomalies in HPA axis function in adulthood . Clinically, it may be postulated that disturbed fetal brain development due to prenatal immune challenge increases the vulnerability to develop mental illness involving inadequate responses to stress. Surg Clin North Am, 1994 Jun, 74(3), 497 - 517 Principles of antibiotic therapy; Solomkin JS et al.; The primary development in the area of antibiotic treatment for surgical infections in the last 5 years has been the expanded clinical importance of beta-lactamases in the protection of Gram-negative organisms from previously active drugs . To counter this problem, a series of new antibiotic agents has been developed, including new cephalosporins, carbapenems, quinolones, and beta-lactamase inhibitors . This article describes the various beta-lactamases and their mechanisms of action, and details the activity of new antibiotic agents against resistant Gram-negative organisms . Recent information on the importance of combination therapy for patients with severe Gram-negative infections is reviewed . The use of optimized aminoglycoside dosing regimens, including once-a-day dosing, provides an additional strategy for treating serious Gram-negative infections. J Immunol, 1994 Jun 1, 152(11), 5420 - 8 Regulation of neutrophil responses by phosphotyrosine phosphatase; Cui Y et al.; By using immunofluorescent flow cytometry, we observed a profound up-regulation of CD45 on the plasma membrane of neutrophils exposed to low levels of a culture supernatant of the Gram-negative pathogen, Fusobacterium nucleatum (FN) . Plasma membranes of neutrophils freshly prepared form human blood possessed little enzymatically active phosphotyrosine phosphatase . The activity of this enzyme was markedly potentiated in plasma membranes prepared from cells preexposed to the FN culture supernatant . This activity was vanadate sensitive and could be immunoprecipitated with anti-CD45 Ab . Cells preexposed to the FN culture supernatant were inhibited in their ability to release superoxide when challenged with the bacterial chemotactic factor, FMLP, but not PMA . The tyrosine kinase inhibitor, genistein, likewise inhibited FMLP but not PMA-induced superoxide release . Pretreatment of neutrophils with vanadate reversed FN-mediated inhibition of FMLP-triggered superoxide release but had no effect on genistein-mediated inhibition of FMLP-induced superoxide release . Of several proteins tyrosine phosphorylated in response to treatment of neutrophils with FMLP, Western analysis revealed one (m.w . approximately 93,000) that was lost when FMLP-stimulated cells were exposed to FN . This effect was inhibited when the cells were preexposed to vanadate . These results are consistent with the hypothesis that plasma membrane tyrosine phosphatase modulates FMLP-induced superoxide release by reversing the effects of tyrosine kinases activated in the initial phases of cell stimulation. Infect Immun, 1994 Jun, 62(6), 2315 - 21 Uroepithelial cells are part of a mucosal cytokine network; Hedges S et al.; This study compared the cytokine production of uroepithelial cell lines in response to gram-negative bacteria and inflammatory cytokines . Human kidney (A498) and bladder (J82) epithelial cell lines were stimulated with either Escherichia coli Hu734, interleukin 1 alpha (IL-1 alpha), or tumor necrosis factor alpha (TNF-alpha) . Supernatant samples were removed, and the RNA was extracted from cells at 0, 2, 6, and 24 h . The secreted cytokine levels were determined by bioassay or immunoassay; mRNA was examined by reverse transcription-PCR . The two cell lines secreted IL-6 and IL-8 constitutively . IL-6 and IL-8 mRNA were constitutively produced in both cell lines; IL-1 beta mRNA was detected in J82 cells . IL-1 alpha induced significantly higher levels of IL-6 secretion than did E . coli Hu734 or TNF-alpha . IL-1 alpha and TNF-alpha induced significantly higher levels of IL-8 secretion than did E . coli Hu734 . Secreted IL-1 beta was not detected; IL-1 alpha and TNF-alpha were not detected above the levels used for stimulation . IL-1 alpha, IL-1 beta, IL-6, and IL-8 mRNAs were detected in both cell lines after exposure to the stimulants . TNF-alpha mRNA was occasionally detected in the J82 cell line after TNF-alpha stimulation . Cytokine (IL-6 and IL-8) and control (glyceraldehyde 3-phosphate dehydrogenase {G3PDH} and beta-actin) mRNA concentrations were quantitated with internal PCR standards . Cytokine mRNA levels relative to beta-actin mRNA levels were the highest in E . coli-stimulated cells . In comparison, the cytokine mRNA levels relative to G3PDH mRNA levels were the highest in IL-1 alpha-stimulated cells . beta-Actin mRNA levels decreased after bacterial stimulation but not after cytokine stimulation, while G3PDH mRNA levels increased in response to all of the stimulants tested . These results suggested that E . coli Hu734 lowered the beta-actin mRNA levels in uroepithelial cells, thus distorting the IL-6 and IL-8 mRNA levels relative to this control . In summary, E . coli IL-1 alpha and TNF-alpha were found to activate the de novo synthesis and secretion of IL-6 and IL-8 in uroepithelial cells . These results emphasize the role of epithelial cells in cytokine-mediated responses during the early stages of infection. Infect Immun, 1994 Jun, 62(6), 2270 - 6 Lipopolysaccharide smooth-rough phase variation in bacteria of the genus Chlamydia; Lukacova M et al.; In two strains of Chlamydia psittaci and in Chlamydia trachomatis serotype L1, we have detected a so-far-unknown antigen which (i) is resistant to heat and proteolytic digestion, (ii) can be extracted with phenol-water into the water phase, (iii) gives a ladder-like banding pattern in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, (iv) is immunogenic in rabbits and mice, and (v) contains immunoreactivity of lipid A, a common and characteristic component of gram-negative lipopolysaccharides (LPS) . Thus, chlamydiae contain, in addition to the known rough-type LPS, another LPS type which is phenotypically smooth (S-LPS) . S-LPS was observed preferentially in chlamydiae grown in the yolk sac of embryonated eggs; it was, however, also detected by immunofluorescence in tissue culture-grown chlamydiae with a monoclonal antibody against S-LPS. Trends Microbiol, 1994 Jun, 2(6), 193 - 8 Gram-negative bacterial communication by N-acyl homoserine lactones: a universal language? Swift S, Bainton NJ, Winson MK. The ability of bacterial cells to use small signalling molecules to monitor population growth may help them to react rapidly to environmental change . Regulatory systems made up of a small sensor molecule, an N-acyl homoserine lactone and a protein effector have been identified recently in a wide range of Gram-negative bacteria . These mediate signal cascades that amplify and coordinate the induction of single or multiple regulons. J Appl Bacteriol, 1994 Jun, 76(6), 559 - 67 Carbofuran-degrading bacteria from previously treated field soils; Parekh NR et al.; Laboratory incubation studies were made on soils collected from five field sites with different histories of treatment with carbofuran . All soils treated earlier with carbofuran degraded the compound more rapidly than untreated samples of the same soils . Reduced rates of degradation in the presence of chloramphenicol imply that soil bacteria are primarily responsible for the breakdown of carbofuran in these soils . Sixty-eight bacteria, capable of degrading carbofuran as the sole source of carbon and nitrogen, were isolated from liquid cultures of treated soils . The concentration of carbofuran in the liquid medium used for isolation and subsequent culture of carbofuran-degrading isolates appeared to affect the stability of their ability to degrade . Similar types of carbofuran-degrading bacteria were isolated from different soils and several different types were isolated from one soil . All carbofuran-degrading isolates were Gram-negative, aerobic rods which hydrolysed the insecticide to carbofuran phenol . They were separated into four groups on the basis of a limited number of phenotypic characters . There was a good correlation between the phenotype of carbofuran-degrading isolates and the stability of their ability to degrade . Fourteen isolates were placed in phenotypic group I and 13 of these did not degrade carbofuran after one subculture in liquid medium . Phenotypic groups II, III and IV consisted of 54 isolates in total (3, 46 and 5 isolates respectively) and 52 of these retained their ability to degrade carbofuran when subcultured. J Surg Res, 1994 Jun, 56(6), 579 - 85 Polymicrobial sepsis but not low-dose endotoxin infusion causes decreased splenocyte IL-2/IFN-gamma release while increasing IL-4/IL-10 production; Ayala A et al.; Although studies indicate that polymicrobial sepsis produces marked depression in lymphocyte functions, it remains unclear whether this dysfunction is due to the chronic exposure of immune cells to endotoxin (ETX; a product of the gram-negative bacterial cell wall) at levels typically encountered in the septic state . The aim of this study, therefore, was to determine whether the changes in lymphokine release seen during polymicrobial sepsis are comparable to those observed with chronic ETX infusion . To assess this, splenocytes were harvested from C3H/HeN mice (ETX-sensitive) at 1 or 24 hr following cecal ligation and puncture (CLP; to induce polymicrobial sepsis), Sham CLP (Sham), or laparotomy followed by peritoneal implantation of a mini-osmotic pump which delivered either saline vehicle (Sal-pump) or ETX (ETX-pump; 0.025 micrograms lipopolysaccharide/25 g body wt/24 hr) . Splenocytes were then stimulated with concanavalin A (2.5 micrograms/ml/48 hr) and their capacity to release interleukin (IL)-2, interferon (IFN)-gamma, IL-4, and IL-10 was determined by bioassay or ELISA . The results indicated that there were no changes in lymphokine release capacity at 1 hr after CLP or ETX-pump implantation . However, prolonged sepsis (i.e., at 24 hr) caused a marked suppression of IL-2 and IFN-gamma release (immune-enhancing lymphokines characteristic of Th1-cells), while enhancing the release of immunosuppressive Th2-cell products IL-4 and IL-10 . Chronic exposure to ETX at a level comparable to that seen in CLP caused no depression in lymphokine (IL-2/IFN-gamma) release . This implies that a bacterial component other than ETX mediates the differential alterations observed in lymphokine release during prolonged polymicrobial sepsis. J Surg Res, 1994 Jun, 56(6), 549 - 55 Tissue tumor necrosis factor mRNA expression following cecal ligation and puncture or intraperitoneal injection of endotoxin; Hadjiminas DJ et al.; Tumor necrosis factor (TNF) has been implicated as a key mediator of the septic response . Although very high serum levels of TNF are detected in animal models of endotoxemia or gram-negative bacteremia, human patients with sepsis rarely have greatly elevated TNF serum levels . It has therefore been postulated that TNF may act in a paracrine fashion to cause local injury . In order to examine the role of locally produced TNF in sepsis, we compared serum TNF levels and TNF messenger RNA (mRNA) expression in various tissues following cecal ligation and puncture (CLP) or intraperitoneal injection of a sublethal dose of endotoxin . TNF mRNA expression was determined by the reverse transcription differential polymerase chain reaction using beta-actin as an internal standard . Serum levels of TNF were threefold higher after endotoxin administration compared to CLP . TNF mRNA in peritoneal macrophages rose fourfold after both endotoxin injection and CLP, with rapid return to normal in endotoxin-treated animals . There was a significant increase in TNF mRNA in the liver and lung, but not the spleen, during the first 24 hr after CLP . An increase in TNF mRNA was seen in all three tissues after injection of endotoxin . These results support the concept of locally produced TNF as a contributing factor in tissue damage and multiple organ failure during sepsis. Lab Invest, 1994 Jun, 70(6), 862 - 7 Lack of suppressed renal thrombomodulin expression in a septic rat model with glomerular thrombotic microangiopathy; Laszik Z et al.; BACKGROUND: The thrombomodulin-dependent protein C anticoagulant pathway plays a major physiologic role in the down-regulation of the coagulation process . In cell culture, inflammatory cytokines or endotoxin can down-regulate endothelial thrombomodulin (TM) suggesting that suppressed TM expression may contribute to thrombotic complications noted in Gram-negative sepsis . EXPERIMENTAL DESIGN: In the present study, we have examined TM expression in the kidneys of septic rats utilizing indirect immunofluorescence and have quantified TM antigen and TM activity in extracts of the same kidneys by enzyme-linked immunosorbent assays and protein C activation assays, respectively . Conscious Sprague-Dawley rats were injected intravenously with LD95 doses of live E . coli (N = 30), or endotoxin (N = 30) . Control animals (N = 30) were injected with equivalent volumes of saline . The rats were killed 30, 90, 180, 360, and 720 minutes after the initiation of sepsis . RESULTS: Glomerular capillary thrombosis developed by 180 minutes in approximately half of the animals after the initiation of sepsis . We failed to demonstrate suppressed TM expression in the kidneys of septic animals using immunofluorescence . Neither enzyme-linked immunosorbent assays, nor protein C activation assays showed decreased levels in TM antigen expression or activity at different time points during the sepsis . CONCLUSIONS: These results indicate that suppressed TM expression does not contribute to the development of the glomerular capillary thrombosis in this septic rat model. Eur J Surg, 1994 Jun-Jul, 160(6-7), 363 - 7 Bacteriological studies of bile from the gallbladder in patients with carcinoma of the gallbladder, cholelithiasis, common bile duct stones and no gallstones disease; Csendes A et al.; OBJECTIVE: To compare the presence of bacteria of bile from the gallbladder in control subjects, patients with gallstones, and patients with carcinoma of the gallbladder . DESIGN: Prospective open study . SETTING: University department of surgery . SUBJECTS: 372 patients of whom 36 had no signs of gallbladder disease; 211 underwent cholecystectomy for either symptomatic gallstone disease (n = 165) or acute cholecystitis (n = 46); 67 had common bile duct stones and 58 were operated on for carcinoma of the gallbladder . INTERVENTIONS: Aspiration of bile from the gallbladder . MAIN OUTCOME MEASURES: Prevalence of pathogenic bacteria in bile from the gallbladder and correlations between the presence of bacteria, the presence of cancer, and age . RESULTS: No pathogenic bacteria were grown from the bile of the patients who had no signs of gallstones disease . Among the 165 with symptomatic gallstone disease 52 (32%) had pathogens in their bile, and among the 46 with acute cholecystitis the corresponding figure was 19 (41%) while among patients with common bile duct stones this figure was 39 (58%) . Among the 58 patients with carcinoma of the gallbladder the bile grew organism in 47 (81%) . Patients over the age of 60 years tended to be more likely to have organism in their bile than patients aged 60 or less, and the difference was significant for symptomatic gallstone disease (p < 0.003) . Significant differences were also found between patients with symptomatic gallstone disease and those with carcinoma of the gallbladder in both age groups (p < 0.002 in each case) . Most of the organism were aerobic or anaerobic Gram negative species, irrespective of type of disease or age . CONCLUSION: Bacteria may have a role in the development of carcinoma of the gallbladder. Eur Respir J, 1994 Jun, 7(6), 1125 - 30 Niacin attenuates acute lung injury induced by lipopolysaccharide in the hamster; Nagai A et al.; Lipopolysaccharide plays a major role in the development of lung injury induced by Gram-negative bacteria, but a protective agent to attenuate the LPS-induced lung injury has not been found . The aim of this study was to examine the effects of niacin on LPS-induced acute lung injury . We administered LPS (Escherichia coli) 0.01 mg.100 g-1 body weight, transtracheally into the lungs of hamsters . Niacin (250 or 500 mg.kg-1 body weight) was injected intraperitoneally 24 h before, and 1 h after the LPS administration . LPS treatment increased wet/dry lung weight, albumin content and neutrophil counts in bronchoalveolar lavage fluid . In hamsters treated with niacin, wet/dry lung weight, albumin content and intra-alveolar cell counts were normal . Nicotinamide adenine dinucleotide (NAD) was significantly decreased in lung tissue of hamsters treated with LPS alone, but was increased in hamsters treated with LPS and niacin . Histopathological examination revealed that niacin-treated LPS-administered hamsters had lungs with no or occasional inflammatory cell infiltration in alveolar spaces, in contrast to the lungs of LPS-treated hamsters, which were infiltrated with numerous inflammatory cells . We conclude that niacin attenuates LPS-induced acute lung injury, probably, in part, by preventing the depletion of NAD. Ann Pharmacother, 1994 Jun, 28(6), 757 - 66 Once-daily administration of aminoglycosides; Bates RD et al.; OBJECTIVE: To provide an overview of the efficacy and adverse effects associated with once-daily administration of aminoglycosides . DATA SOURCES: An extensive MEDLINE search and review of journals was conducted to identify information for this review . DATA SYNTHESIS: Aminoglycosides alone or in combination with beta-lactams are commonly used for their activity against gram-negative microorganisms . Numerous studies have been performed comparing efficacy and toxicity of once-daily administration of aminoglycosides with multiple-daily dosing . Two studies have found a significant difference in clinical efficacy between once-daily and multiple-daily dosing of aminoglycosides . Several studies have observed a lower incidence of toxicity with once-daily than multiple-daily dosing, but others have found no difference . CONCLUSIONS: Review of the literature suggests that once-daily administration of aminoglycosides may be as safe and effective as multiple-daily dosing regimens for the treatment of certain infections caused by gram-negative bacteria. Neurosurgery, 1994 Jun, 34(6), 974 - 80; discussion 980-1 Intracranial suppuration: a modern decade of postoperative subdural empyema and epidural abscess; Hlavin ML et al.; A retrospective study of subdural empyema and epidural abscess spanning 11 years and encompassing 41 patients was performed, demonstrating that the clinical characteristics of intracranial suppuration have changed over time . Sinusitis and otitis media, previously the predominant etiologies, were predisposing factors in only 29% of patients . A prior craniotomy had been performed in 66% of cases and was the most common risk factor for abscess development . The postoperative patients were subjected to detailed analysis . Patients who had undergone a prior craniotomy were notable for the following features: older age, lack of fever, evidence of wound infection, frequent false-negative computed tomographic scans, and a high percentage of Gram negative aerobic organisms or skin flora as pathogens . The population at highest risk for abscess development ranged from 50 to 60 years old, older than in previous series . Older age and an advanced degree of encephalopathy were indicative of a poor prognosis . Patients with subdural empyema had a worse prognosis as well . Hyponatremia was a frequent complicating factor . A much greater percentage of Gram-negative aerobic bacteria were isolated than in previous studies . Computed tomographic scans, half of which were performed with intravenous contrast material, were nondiagnostic in 30% of patients . The mortality rate was 18.5%, and delay in treatment correlated with increased risk of poor outcome . All patients were treated with a craniotomy . Repeated operations were required in three patients and were associated with the development of intraparenchymal abscess. Shock, 1994 Jun, 1(6), 432 - 7 Effects of sepsis on recovery of the heart from 50 min ischemia; McDonough KH et al.; Gram-negative sepsis as well as administration of agents that simulate or occur naturally subsequent to a septic challenge, can present as an oxidant stress to the myocardium . These stresses may then induce the development of protection of the heart from future stresses . This protection of the heart may occur in spite of the fact that sepsis itself induces myocardial dysfunction . In the present study we determined if sepsis is protective of a 50 min ischemic episode, one in which some degree of irreversible damage may occur . In addition we determined if sepsis-induced protection was still present when this ischemic challenge was imposed upon the heart of the alcoholic septic animal in which the chronic alcoholic state can lead to a potentiation of sepsis induced cardiac depression . Thus animals were fed an ethanol containing diet or a control diet for 8-10 weeks and were then made septic by the administration of Escherichia coli into the dorsal subcutaneous space . Control animals received sterile saline . The following day, hearts were studied in the isovolumic beating preparation and, after basal function was assessed, hearts were made globally ischemic for 50 min and reperfused for 30 min . Left ventricular pressure was continuously monitored and coronary flow was measured at specific intervals . After ischemia and reperfusion, hearts from control- and alcohol-fed animals that were nonseptic showed significantly decreased left ventricular performance . Ventricular pressure development of hearts from septic and alcoholic septic rats was not significantly decreased after ischemia and reperfusion compared to preischemia although preischemic function was significantly lower in the sepsis groups compared to their nonseptic control groups.(ABSTRACT TRUNCATED AT 250 WORDS) Ann N Y Acad Sci, 1994 May 28, 725, 173 - 82 Endotoxin-induced neutrophil adherence to endothelium: relationship to CD11b/CD18 and L-selectin expression and matrix disruption; Finn A et al.; The injury to vascular endothelium seen in severe bacterial infection may be mediated by neutrophil-derived enzymes . Neutrophil adhesion to endothelium, a prerequisite for this process, is mediated sequentially by the leukocyte adhesion molecules L-selectin and the beta 2 integrins, including CD11b/CD18 . We have explored the relationship between expression of these molecules, neutrophil adherence, endothelial activation, and consequent endothelial injury, as assessed in vitro by changes to HS and FN matrices that colocalize . Endothelial prestimulation with LPS (endotoxin) caused an increase in adherence and an inversely proportional disruption in the HS matrix; disruption of the FN matrix only occurred on the further addition of fMLP . Although maximal changes in these matrices were associated with elevation of neutrophil CD11b/CD18 and reduction in L-selectin expression, these changes did not determine either the nature or extent of endothelial damage . CD11b/CD18 expression was similar in both adherent and nonadherent neutrophils, while L-selectin was shed in association with adherence in the absence of other stimuli . These changes in expression were thus independently regulated . This model may provide further insights into the interrelationship between neutrophil adhesion and activation and endothelial damage in infection with gram-negative bacteria. Blood, 1994 May 15, 83(10), 2985 - 94 The heterogeneity of azurophil granules in neutrophil promyelocytes: immunogold localization of myeloperoxidase, cathepsin G, elastase, proteinase 3, and bactericidal/permeability increasing protein; Egesten A et al.; Azurophil granules of myeloid cells form in promyelocytes . They store cytotoxic and digestive agents which when released are involved in the defense against infection . In order to characterize the intragranular distribution of these agents, ultrastructural methods using immunogold were used on promyelocytes . Azurophil granules were divided into nucleated, large spherical (large azurophil) and small electron-dense (small azurophil) granules . Myeloperoxidase showed a peripheral distribution of large azurophils and a uniform distribution of small and nucleated azurophils, consistent with previous findings . Likewise, the major neutral proteases of azurophils, cathepsin G, granulocyte elastase, and proteinase 3, displayed a similar distribution, with a peripheral localization in large azurophils and a uniform distribution in small and nucleated azurophils, except for proteinase 3, which was associated with the crystalloid structure in nucleated azurophils . In contrast, the bactericidal/permeability increasing protein, which is bacteristatic and bactericidal for Gram-negative bacteria, was localized to the membrane area in all types of azurophil granules, consistent with a suggested association of this protein with the granule membrane . The observed differences in intragranular distribution of the proteins investigated may reflect variations in binding to matrix structures and granule membranes. Proc Natl Acad Sci U S A, 1994 May 10, 91(10), 4308 - 12 Structural similarity of a developmentally regulated bacterial spore coat protein to beta gamma-crystallins of the vertebrate eye lens; Bagby S et al.; The solution structure of Ca(2+)-loaded protein S (M(r) 18,792) from the Gram-negative soil bacterium Myxococcus xanthus has been determined by multidimensional heteronuclear NMR spectroscopy . Protein S consists of four internally homologous motifs, arranged to produce two domains with a pseudo-twofold symmetry axis, overall resembling a triangular prism . Each domain consists of two topologically inequivalent "Greek keys": the second and fourth motifs form standard Greek keys, whereas the first and third motifs each contain a regular alpha-helix in addition to the usual four beta-strands . The structure of protein S is similar to those of the vertebrate eye lens beta gamma-crystallins, which are thought to be evolutionarily related to protein S . Both protein S and the beta gamma-crystallins function by forming stable multimolecular assemblies . However, protein S possesses distinctive motif organization and domain packing, indicating a different mode of oligomerization and a divergent evolutionary pathway from the beta gamma-crystallins. J Infect Dis, 1994 May, 169(5), 1151 - 4 Circulating polymorphonuclear leukocytes from patients with gram-negative bacteremia are not primed for enhanced production of leukotriene B4 or 5-hydroxyeicosatetraenoic acid; Sorrell TC et al.; The hypothesis was tested that polymorphonuclear leukocytes (PMNL) from patients with gram-negative bacteremia are primed to produce leukotriene B4 (LTB4) or 5-hydroxyeicosatetraenoic acid (5-HETE), in response to concentrations of calcium ionophore A23187, which are substimulatory for control PMNL . PMNL from 11 bacteremic patients and 8 healthy subjects (11 samples) produced similar quantities of LTB4, omega-oxidation products of LTB4, and 5-HETE after incubation with 0.3 and 0.5 microM A23187 for 5 min . At the detection threshold of 0.3 microM A23187, LTB4 was present in PMNL preparations from 9 of 11 patients and 7 of 11 control samples and 5-HETE from the same 9 patients and from 6 controls . There was no correlation between LTB4 or 5-HETE and plasma levels of endotoxin . In this group of patients, priming of PMNL by gram-negative bacteremia did not lead to enhanced production of LTB4, its omega-oxidation products, or 5-HETE when PMNL were challenged with low concentrations of A23187. J Exp Med, 1994 May 1, 179(5), 1653 - 58 Gene structure of the Helicobacter pylori cytotoxin and evidence of its key role in gastric disease; Telford JL et al.; The gram negative, microaerophilic bacterium Helicobacter pylori colonizes the human gastric mucosa and establishes a chronic infection that is tightly associated with atrophic gastritis, peptic ulcer, and gastric carcinoma . Cloning of the H . pylori cytotoxin gene shows that the protein is synthesized as a 140-kD precursor that is processed to a 94-kD fully active toxin . Oral administration to mice of the purified 94-kD protein caused ulceration and gastric lesions that bear some similarities to the pathology observed in humans . The cloning of the cytotoxin gene and the development of a mouse model of human gastric disease will provide the basis for the understanding of H . pylori pathogenesis and the development of therapeutics and vaccines. Ann Intern Med, 1994 May 1, 120(9), 771 - 83 Selected treatment strategies for septic shock based on proposed mechanisms of pathogenesis; Natanson C et al.; PURPOSE: To review selected new therapies for septic shock designed to inhibit bacterial toxins or endogenous mediators of inflammation . DATA SOURCES: Scientific journals, scientific meeting proceedings, and Food and Drug Administration advisory committee proceedings . STUDY SELECTION AND EXTRACTION: Preclinical and clinical data from trials using core-directed antiendotoxin antibodies and anticytokine therapies for sepsis and studies in animal models of sepsis from our laboratory . RESULTS OF DATA SYNTHESIS: Ten clinical trials using core-directed antiendotoxin antibodies produced inconsistent results and did not conclusively establish the safety or benefit of this approach . Both anti-interleukin-1 and anti-tumor necrosis factor (TNF) therapies have been beneficial in some animal models of sepsis but did not clearly improve survival in initial human trials, and one anti-TNF therapy actually produced harm . Neutrophils, another target for therapeutic intervention, protect the host from infection but may also contribute to the development of tissue injury during sepsis . In a canine model of septic shock, granulocyte colony-stimulating factor increased the number of circulating neutrophils and improved survival, but an anti-integrin (CD11/18) antibody that inhibits neutrophil function worsened outcome . Nitric oxide, a vasodilator produced by the host, causes hypotension during septic shock but may also protect the endothelium and maintain organ blood flow . In dogs challenged with endotoxin, the inhibition of nitric oxide production decreased cardiac index and did not improve survival . CONCLUSIONS: No new therapy for sepsis has shown clinical efficacy . Perhaps more accurate clinical and laboratory predictors are needed to identify patients who may benefit from a given treatment strategy . On the other hand, the therapeutic premises may be flawed . Targeting a single microbial toxin such as endotoxin may not represent a viable strategy for treating a complex inflammatory response to diverse gram-negative bacteria . Similarly, the strategy of inhibiting the host inflammatory response may not be beneficial because immune cells and cytokines play both pathogenic and protective roles . Finally, our scientific knowledge of the complex timing of mediator release and balance during sepsis may be insufficient to develop successful therapeutic interventions for this syndrome. Antimicrob Agents Chemother, 1994 May, 38(5), 959 - 62 beta-Lactamase hydrolysis of cephalosporin 3'-quinolone esters, carbamates, and tertiary amines; Georgopapadakou NH et al.; The beta-lactam hydrolysis of five cephalosporin 3'-quinolones (dual-action cephalosporins) by three gram-negative beta-lactamases was examined . The dual-action cephalosporins tested were the ester Ro 23-9424; the carbamates Ro 25-2016, Ro 25-4095, and Ro 25-4835; and the tertiary amine Ro 25-0534 . Also tested were cephalosporins with similar side chains (cefotaxime, desacetylcefotaxime, cephalothin, cephacetrile, and Ro 09-1227 {SR 0124}) and standard beta-lactams (penicillin G, cephaloridine) . The beta-lactamases used were the plasmid-mediated TEM-1 and TEM-3 enzymes and the chromosomal AmpC . The cephacetrile-related compounds Ro 25-4095 and Ro 25-4835 were hydrolyzed by all three beta-lactamases with catalytic efficiencies (relative to penicillin G) ranging from approximately 5 (TEM-1, AmpC) to approximately 25 (TEM-3) . The cephalothin-related Ro 25-2016 was also hydrolyzed by all three beta-lactamases, particularly the AmpC enzyme (relative catalytic efficiency, 110) . The cefotaxime-related compounds Ro 25-0534 and Ro 23-9424 were hydrolyzed to any significant extent only by the TEM-3 enzyme (relative catalytic efficiencies, 1.2 and 4.7, respectively. Mol Gen Mikrobiol Virusol, 1994 May-Jun, (3), 3 - 9 {Resistance to metals determined by plasmids of gram-negative bacteria}; Anisimova LA et al.; The data are reviewed on plasmid determinants controlling resistance of gram-negative bacteria to heavy metals . The structural and functional arrangement of operons determining resistance to copper, zinc, cadmium, cobalt, nickel, tellurites, and chromates is emphasized . The fields of use of the genetic potential of the bacteria resistant to metals in the modern biotechnological processes are envisaged. J Vet Intern Med, 1994 May-Jun, 8(3), 203 - 6 Hepatic abscesses associated with diabetes mellitus in two dogs; Grooters AM et al.; Two diabetic dogs were presented for anorexia, persistent fever, and poor control of hyperglycemia . Both had neutrophilia with left shift, hypoalbuminemia, and increased serum alkaline phosphatase (SAP) activity . Radiography indicated intrahepatic gas densities in 1 dog and a hepatic mass in the other . Abdominal sonography demonstrated multiple well-demarcated hypoechoic hepatic lesions consistent with abscesses . Both dogs were successfully treated by surgical resection of the abscessed liver lobes in conjunction with antibiotics and supportive therapy . Good control of hyperglycemia was achieved in both dogs after recovery . Intracellular and extracellular Gram-negative rod-shaped bacteria were abundant in the abscesses from both dogs . These cases suggest an association between diabetes mellitus and hepatic abscessation. J Appl Physiol, 1994 May, 76(5), 2006 - 14 Bactericidal/permeability-increasing protein ameliorates acute lung injury in porcine endotoxemia; Vandermeer TJ et al.; Bactericidal/permeability-increasing protein (BPI), a cationic protein isolated from human neutrophils, binds lipopolysaccharide (LPS), kills gram-negative bacteria, and neutralizes many of the effects of LPS in vitro and in vivo . We hypothesized that a recombinant 23-kDa NH2-terminal fragment of BPI (BPI23) would reduce acute lung injury in endotoxemic pigs . At -18 h, pigs received an intravenous priming dose of LPS (20 micrograms/kg) . Anesthetized ventilated swine were randomized to receive 1) no further treatment (n = 4); 2) LPS (250 micrograms/kg over 50 min) and BPI23 (3-mg/kg bolus and 3 mg/kg over 60 min) (n = 6); or 3) LPS and thaumatin, a cationic protein devoid of LPS neutralizing activity that has a molecular mass and isoelectric point that are similar to that of BPI23 (n = 7) . BPI23 treatment significantly ameliorated LPS-induced hypoxemia, functional upregulation of opsonin receptors on circulating phagocytes, and alveolitis but had no effect on the elaboration of tumor necrosis factor-alpha or thromboxane A2 . The salutory effects of BPI23 on acute lung injury in endotoxemic pigs may be mediated, at least in part, by inhibition of direct activation of phagocytes by LPS. J Am Vet Med Assoc, 1994 May 1, 204(9), 1475 - 8 Clinical ehrlichiosis in a cat; Bouloy RP et al.; Clinical ehrlichiosis was diagnosed in a cat from Colorado on the basis of cytologic, serologic, and clinical findings . Clusters of gram-negative organisms that were morphologically similar to morulae of Ehrlichia spp were found only in mononuclear cells . The cat had clinical signs that were referable to infection by a rickettsial organism, and antibodies against E canis (titer, 1:80) and E risticii (titer, 1:40) were detected in serum . Exclusion of other obvious causes inducing similar clinical signs, and positive clinical response to doxycycline, an antibiotic with known antirickettsial actions, aided in diagnosis . The cat was clinically normal within days following initiation of treatment, and was clinically normal, as well as seronegative for antibodies against E canis and E risticii, 1,365 days after discharge. FEMS Microbiol Lett, 1994 May 1, 118(1-2), 113 - 9 Accumulation and intracellular fate of tellurite in tellurite-resistant Escherichia coli: a model for the mechanism of resistance; Lloyd-Jones G et al.; The tellurite accumulation properties of three Escherichia coli strains containing different tellurium-resistance determinants of Gram-negative origin, from plasmids pMER610, pHH1508a and RK2, were compared . In all three cases membrane-associated tellurium crystallization was observed, and neither reduced uptake nor increased export contributed to the resistance . Specific membrane-proximal reduction is proposed as the mechanism of resistance to tellurite coded by all three determinants, despite their lack of sequence homology. Res Microbiol, 1994 May, 145(4), 269 - 72 Fever and the control of gram-negative bacteria; Green MH et al.; Although it seems obvious that fever has some important general adaptive value, it is still not clear by what means this function is manifested . One postulate is that febrile conditions result in the sequestration of soluble iron, effectively starving some pathogens of that essential nutrient . On the basis of our recent experiments, we propose a new mechanism for how fever serves to restrict a wide range of Gram-negative bacteria . The elevated temperature prevents the bacteria from synthesizing their protective LPS, thereby enabling serum complement to perforate and kill the invading pathogens even prior to the production of host antibodies. Circ Shock, 1994 May, 43(1), 34 - 43 Altered mitochondrial redox responses in gram negative septic shock in primates; Simonson SG et al.; Gram negative sepsis causes changes in oxygen supply-demand relationships . We have used a primate model of hyperdynamic gram negative sepsis produced by intravenous infusion of Escherichia coli (E . coli) to evaluate sepsis-induced alterations in mitochondrial oxidation-reduction (redox) state in muscle in vivo . The redox state of cytochrome a,a3, the terminal member of the intramitochondrial respiratory chain, was assessed in the intact forearm by near-infrared (NIR) spectroscopy . The muscle NIR data were compared to routine measures of oxygen delivery (DO2) and oxygen consumption (VO2) . After E . coli infusion and fluid resuscitation, DO2 and VO2 showed minimal changes through 24 hr of sepsis . In contrast, changes in cytochrome a,a3 redox state evaluated by NIR occurred within a few hours and were progressive . Mitochondrial functional responses were correlated with structural changes observed on serial muscle biopsies . Gross morphological changes in muscle mitochondria were present in some animals as early as 12 hr, and, in most animals, by 24 hr . The morphologic changes were consistent with decreases in oxidative capacity as suggested by NIR spectroscopy . The NIR data also suggest that two mechanisms are operating to explain abnormalities in oxygen metabolism and mitochondrial function in lethal sepsis . These mechanisms include an early defect in oxygen provision to mitochondria that is followed by a progressive loss in functional cytochrome a,a3 in the muscle. FEMS Immunol Med Microbiol, 1994 May, 8(4), 321 - 8 Heat shock response in Actinobacillus actinomycetemcomitans; Lokensgard I et al.; The heat shock response in Actinobacillus actinomycetemcomitans, a capnophilic Gram-negative bacterial species that is implicated in the development of certain forms of periodontitis, was characterized . Different strains of A . actinomycetemcomitans were grown at 37, 42 and 48 degrees C in the presence of 35S-methionine . The bacterial cells were lysed, run on SDS-PAGE and subsequently blotted on nitrocellulose paper . After autoradiography of the blots, several protein bands from the cultures at 42 degrees C showed an increased intensity; major bands were observed at 90, 70, and 60 kDa, but increased protein synthesis was also detected at 54, 28 and 17 kDa . Nitrocellulose blots were also incubated with a panel of monoclonal and polyclonal antibodies directed to epitopes on different heat shock proteins . Strong reactivity was found with several antibodies at the position corresponding to a molecular mass of 60 kDa . The protein is probably the GroEL homologue in A . actinomycetemcomitans, a member of the 'common bacterial antigen' family. Mol Cell Biol, 1994 May, 14(5), 2914 - 25 Definition of a lipopolysaccharide-responsive element in the 5'-flanking regions of MuRantes and crg-2; Shin HS et al.; Macrophages are stimulated by lipopolysaccharide (LPS) of gram-negative organisms . The changes in LPS-stimulated macrophages include transcriptional activation of multiple immediate-early genes, which may contribute to the natural immunity to microorganisms . We have defined by deletion and mutational analysis LPS-responsive elements (LREs) in two chemokine genes, MuRantes and crg-2, which are activated in an immediate-early manner . LRE consists of two motifs, TCAYR, which is an AP-1 half site with two flanking bases, and (A/T) (G/C)NTTYC(A/T)NTTY, which resembles in part the interferon-stimulated responsive element (ISRE) . The orientation of these two motifs relative to each other in MuRantes differed from that in crg-2 . These two motifs are separated by 10 and 6 nonconsensus nucleotides in the MuRantes and crg-2 LREs, respectively . Stimulation of macrophage-like RAW 264.7 cells with alpha/beta interferon did not activate MuRantes, indicating that the ISRE-like motif in MuRantes does not have ISRE activity . Upon stimulation of RAW 264.7 cells with LPS, proteins capable of binding to LRE accumulate in the nuclei as measured by electrophoretic mobility shift assay . These LRE-binding proteins include c-Jun and CREB. Biochem Pharmacol, 1994 Apr 29, 47(9), 1553 - 9 Protective effect of a recombinant fragment of bactericidal/permeability increasing protein against carbohydrate dyshomeostasis and tumor necrosis factor-alpha elevation in rat endotoxemia; Lin Y et al.; Endotoxin (lipopolysaccharide, LPS), a component of the gram-negative bacterial cell wall, induces carbohydrate dyshomeostasis and the release of proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) when administered to experimental animals . Bactericidal/permeability increasing protein (BPI), a cationic protein found in human neutrophil granules, binds with high affinity to LPS and is capable of neutralizing its biological activity . The present study was designed to determine if a recombinant N-terminal fragment of BPI, rBPI23, attenuates LPS-induced alterations in serum glucose, lactate, and TNF-alpha in rats . In anesthetized animals challenged with a 30 min infusion of Escherichia coli O111:B4 LPS (0.25 mg/kg), there was an early transient increase in serum levels of glucose followed by a drop to 60% of those found in saline control rats . A prolonged elevation in serum levels of lactate and a transient, but marked, elevation of TNF-alpha were also observed following LPS infusion . These LPS-induced changes were inhibited significantly by simultaneous infusion of rBPI23 . Different dose-response profiles of rBPI23 on LPS-induced alterations in glucose, lactate and TNF-alpha were observed . When rBPI23 was infused 30 min after the initiation of LPS infusion, it significantly inhibited the alterations in glucose and lactate, but not TNF-alpha . The rise in TNF-alpha was reduced significantly with a 15 min delayed infusion of rBPI23 . A control protein failed to alter any responses to LPS . The results indicate that rBPI23 can provide significant protection against the metabolic disturbances and TNF-alpha release associated with endotoxemia . In addition, the results suggest that LPS-induced metabolic alterations in glucose and lactate are at least partially independent of TNF-alpha release. Biochemistry, 1994 Apr 26, 33(16), 4769 - 79 Refined 1.89-A structure of the histidine-binding protein complexed with histidine and its relationship with many other active transport/chemosensory proteins; Yao N et al.; The structure of the histidine-binding protein (HBP, M(r) = 26,100), involved solely in active transport, has been determined by the molecular replacement technique and refined to 1.89-A resolution and to an R-factor of 0.199 . The structure is that of two protein molecules, each with a bound L-histidine, in the asymmetric unit . Replacement solution was achieved by using a model of the crystal structure of the ligand-free, open-cleft form of the lysine/arginine/ornithine-binding protein which was modified so that the two domains are close to each other by bending the hinge connecting the two domains . The bound histidine is held in place by 10 hydrogen bonds, 2 salt links, and about 60 van der Waals contacts . Elucidation of the HBP structure brings a total of eight different binding proteins structures determined in our laboratory, including those with specificities for monosaccharides, maltodextrins (linear and cyclic), aliphatic amino acids, and inorganic oxyanions . These structures comprise about a third of the entire family of periplasmic binding proteins which act as initial primary high-affinity receptors of active transport in Gram-negative bacteria . Two of the binding proteins with specificities for glucose/galactose and maltodextrins also serve in a similar capacity in chemotaxis . Though these proteins have different molecular weights (ranging from 26,000 to 40,000), amino acid sequences, and ligand specificities, their three-dimensional structures are similar overall . They are elongated (axial ratios of 2:1) and composed of two similar globular domains separated by a deep cleft wherein the ligand-binding site is located . These structures provide understanding of molecular recognition of a variety of ligands at the atomic level and functional roles of the binding proteins. J Biol Chem, 1994 Apr 22, 269(16), 11869 - 72 Functional characterization of the Escherichia coli glycerol facilitator, GlpF, in Xenopus oocytes; Maurel C et al.; The glycerol facilitator of Escherichia coli, GlpF, is a putative nonselective transport channel in the inner membrane of this Gram-negative bacterium . It is a member of the major intrinsic protein (MIP) family of transmembrane channel proteins . Its characterization has been hampered by the lack of a heterologous test system in which its activity can be examined in the absence of other bacterial proteins . Transport of glycerol mediated by this protein was characterized following injection of glpF mRNA into Xenopus laevis oocytes . The properties of GlpF were compared with those of the homologous plant water channel protein, gamma tonoplast intrinsic protein (gamma TIP), as well as the nonhomologous Xenopus K+ channel, Xsha . GlpF selectively transported glycerol but not water or ions, while gamma TIP and Xsha were specific for water and K+, respectively . Voltage clamp experiments showed that GlpF was not voltage-activated for ion transport . Glycerol transport via GlpF proved to be nonsaturable up to 200 mM and exhibited a low temperature of activation (Ea = 4.5 kcal/mol), consistent with the conclusion of Heller et al . (Heller, K . B., Lin, E . C . C., and Wilson, T . H . (1980) J . Bacteriol . 144, 274-278) that GlpF mediates glycerol diffusion via a pore type mechanism . GlpF-mediated transport of glycerol was blocked by mercuric ions (Hg2+) but not N-ethylmaleimide . The inhibitory effect of Hg2+ was partially prevented by inclusion of a high concentration of glycerol and reversed by mercaptoethanol . The results serve to characterize the transport properties of the E . coli glycerol facilitator. Ann N Y Acad Sci, 1994 Apr 15, 712, 146 - 54 C-reactive proteins, limunectin, lipopolysaccharide-binding protein, and coagulin . Molecules with lectin and agglutinin activities from Limulus polyphemus; Liu TY et al.; In 1964, Levin and Bang discovered that gram-negative bacterial endotoxin could rapidly induce gelation of Limulus amebocyte lysate . This observation has led to the development of the most sensitive and specific method for the detection of bacterial endotoxin in pharmaceuticals and drugs intended for human use . Over 10 years ago, Bang injected endotoxin into young horseshoe crabs and observed a time and dose-dependent coagulation of the whole hemolymph . Limunectin, LEBP-PI, and Limulus CRP are found together with coagulin as part of the hemolymph clot at the time of endotoxin-induced exocytosis of amebocytes . In this manner, these molecules with agglutinin/lectin activities could work in concert to assist in the recognition and eventual removal of invading microorganisms from the circulating system . Although the mechanism of endotoxin-induced clot formation is to a large extent understood, the mechanism of clot dissolution and removal in the Limulus hemolymph remains to be clarified. Science, 1994 Apr 15, 264(5157), 418 - 20 Structure of the Tet repressor-tetracycline complex and regulation of antibiotic resistance; Hinrichs W et al.; The most frequently occurring resistance of Gram-negative bacteria against tetracyclines is triggered by drug recognition of the Tet repressor . This causes dissociation of the repressor-operator DNA complex and enables expression of the resistance protein TetA, which is responsible for active efflux of tetracycline . The 2.5 angstrom resolution crystal structure of the homodimeric Tet repressor complexed with tetracycline-magnesium reveals detailed drug recognition . The orientation of the operator-binding helix-turn-helix motifs of the repressor is inverted in comparison with other DNA binding proteins . The repressor-drug complex is unable to interact with DNA because the separation of the DNA binding motifs is 5 angstroms wider than usually observed. Ann Intern Med, 1994 Apr 15, 120(8), 653 - 62 Nosocomial pneumonia in mechanically ventilated patients receiving antacid, ranitidine, or sucralfate as prophylaxis for stress ulcer . A randomized controlled trial; Prod'hom G et al.; OBJECTIVE: To assess three anti-stress ulcer prophylaxis regimens in mechanically ventilated patients for bacterial colonization, early- and late-onset nosocomial pneumonia, and gastrointestinal bleeding . DESIGN: Randomized controlled trial . PATIENTS: Consecutive eligible patients with mechanical ventilation and a nasogastric tube . Of 258 eligible patients, 244 were assessable . SETTING: Medical and surgical intensive care units . INTERVENTION: At intubation, patients were randomly assigned to receive one of the following: antacid (a suspension of aluminum hydroxide and magnesium hydroxide), 20 mL every 2 hours; ranitidine, 150 mg as a continuous intravenous infusion; or sucralfate, 1 g every 4 hours . MEASUREMENTS: Using predetermined criteria, the incidence of gastric bleeding, gastric colonization, early-onset pneumonia, and late-onset pneumonia was assessed in patients intubated for more than 24 hours . RESULTS: Of 244 assessable patients, macroscopic gastric bleeding was observed in 10%, 4%, and 6% of patients assigned to receive sucralfate, antacid, and ranitidine, respectively (P > 0.2) . The incidence of early-onset pneumonia was not statistically different among the three treatment groups (P > 0.2) . Among the 213 patients observed for more than 4 days, late-onset pneumonia was observed in 5% of the patients who received sucralfate compared with 16% and 21% of the patients who received antacid or ranitidine, respectively (P = 0.022) . Mortality was not statistically different among the three treatment groups . Patients who received sucralfate had a lower median gastric pH (P < 0.001) and less frequent gastric colonization compared with the other groups (P = 0.015) . Using molecular typing, 84% of the patients with late-onset gram-negative bacillary pneumonia were found to have gastric colonization with the same bacteria before pneumonia developed . CONCLUSION: Stress ulcer prophylaxis with sucralfate reduces the risk for late-onset pneumonia in ventilated patients compared with antacid or ranitidine. Mol Gen Genet, 1994 Apr, 243(1), 112 - 8 A superfamily of proteins involved in different secretion pathways in gram-negative bacteria: modular structure and specificity of the N-terminal domain; Genin S et al.; The family of PulD proteins, which has been characterized in a wide variety of microorganisms, comprises several membrane-associated proteins essential for the transport of macromolecules across bacterial membranes . These proteins are involved in the transport of complex structures (such as phage particles, DNA) or various proteins (such as extracellular enzymes and pathogenicity determinants) . Amino acid sequence analysis revealed a possible modular organisation of proteins of this superfamily, with highly conserved C-terminal domains and dissimilar N-terminal domains . In the C-terminal domain, four highly conserved regions have been found, one of them containing a remarkable common motif: (V, I)PXL(S, G)XIPXXGXLF . Structural comparisons between the N-terminal domains indicate that proteins of this superfamily can be divided into at least two subgroups, probably reflecting the existence of distinct secretion mechanisms . This implies that members of the superfamily of PulD-related proteins are independently involved in (1) the general secretory pathway, (2) a new signal-peptide-independent secretion pathway found in several bacterial pathogens, and possibly in (3) the translocation of bacteriophage particles through the bacterial cell envelope. Int J Syst Bacteriol, 1994 Apr, 44(2), 204 - 8 Porphyromonas canoris sp . nov., an asaccharolytic, black-pigmented species from the gingival sulcus of dogs; Love DN et al.; A new species, Porphyromonas canoris, is proposed for black-pigmented asaccharolytic strains isolated from subgingival plaque samples from dogs with naturally occurring periodontal disease . This bacterium is an obligately anaerobic, nonmotile, non-spore-forming, gram-negative, rod-shaped organism . On laked rabbit blood or sheep blood agar plates, colonies are light brown to greenish brown after 2 to 4 days of incubation and dark brown after 14 days of incubation . Colonies on egg yolk agar and on nonhemolyzed sheep blood agar are orange . The cells do not grow in the presence of 20% bile and have a guanine-plus-cytosine content of 49 to 51 mol% . The type strain is VPB 4878 (= NCTC 12835) . The average levels of DNA-DNA hybridization between P . canoris strains and other members of the genus Porphyromonas are as follows: Porphyromonas gingivalis ATCC 33277T (T = type strain), 6.5%; Porphyromonas gingivalis cat strain VPB 3492, 5%; Porphyromonas endodontalis ATCC 35406T, 1%; Porphyromonas salivosa NCTC 11362T, 5%; and Porphyromonas circumdentaria NCTC 12469T, 6% . The level of hybridization between P . canoris NCTC 12835T DNA and Porphyromonas asaccharolytica ATCC 25260T DNA is 3% . P . canoris cells produce major amounts of acetic, propionic, isovaleric, and succinic acids and minor amounts of isobutyric and butyric acids as end products of metabolism in cooked meat medium . The major cellular fatty acid is 13-methyltetradecanoic acid (iso-C15:0) . Glutamate and malate dehydrogenases are present, as are glucose-6-phosphate dehydrogenase activity (65.7 nmol mg of protein-1 min-1) and 6-phosphogluconate dehydrogenase activity (63.0 nmol mg of protein-1 min-1).(ABSTRACT TRUNCATED AT 250 WORDS) J Trauma, 1994 Apr, 36(4), 544 - 6; discussion 546-7 Comparison of acid neutralizing and non-acid neutralizing stress ulcer prophylaxis in thermally injured patients; Cioffi WG et al.; We have compared the effectiveness of non-acid neutralizing stress ulcer prophylaxis (SUC) with sucralfate (n = 48) with that of acid neutralizing prophylaxis (AN) utilizing antacids and cimetidine (n = 48) in the prevention of stress ulcer bleeding and nosocomial pneumonia (PN) in thermally injured patients . In the subset of intubated patients, the incidence of PN was 17.8% and 42.8% in the AN and SUC groups, respectively (p < 0.05) despite a similar postburn time of onset of pneumonia . Ten patients in each group died . Three patients in the SUC group developed upper GI bleeding with one requiring gastrectomy . Bacterial colonization of the upper airway occurred in virtually all patients, whereas 83% (SUC) and 96% (AN) had colonization of gastric contents . Gram-negative colonization rates for the upper airway were not different (70%) whereas 48% of SUC patients compared with 60% of AN patients had gram-negative gastric colonization . In conclusion, SUC therapy was efficacious in the prevention of stress ulcer bleeding but did not alter the rate of bacterial colonization of the airway or gastric contents, and was associated with a higher incidence of nosocomial pneumonia in intubated patients. Clin Orthop, 1994 Apr, (301), 205 - 12 Treatment of the infected total hip arthroplasty with a two-stage reimplantation protocol; Lieberman JR et al.; Forty-four patients (46 hips) with infected total hip arthroplasties were evaluated . They were entered into a protocol that included resection arthroplasty, six weeks of intravenous antibiotics which obtained a minimum postpeak serum bactericidal titer of 1:8, and possible reimplantation . Thirty-two of 46 hips (70%) were reimplanted . At an average of 40 months (range, 24-74 months) after reimplantation, infection recurred in three hips (9%) . In two of the three recurrent infections, 1:8 bactericidal titers were not attained . Both of these hips were infected with gram-negative organisms . Minimum postpeak serum bactericidal titers of 1:8 were attained in 28 of 32 hips that were reimplanted, and only one of these hips (4%) had a recurrent infection (p = 0.035) . The presence of retained cement after resection arthroplasty (ten hips) was not associated with recurrent infection . Fourteen hips (12 patients were not reimplanted as a result of a combination of factors, including inadequate bone stock, poor soft-tissue quality, and antibiotic resistance of the infecting organism . The treatment of an infected total hip arthroplasty with resection arthroplasty, six weeks of intravenous antibiotics that attains a minimum postpeak serum bactericidal titer of 1:8, and reimplantation can be an effective and safe treatment. Infect Immun, 1994 Apr, 62(4), 1185 - 91 Competition between rBPI23, a recombinant fragment of bactericidal/permeability-increasing protein, and lipopolysaccharide (LPS)-binding protein for binding to LPS and gram-negative bacteria; Gazzano-Santoro H et al.; Lipopolysaccharide (LPS)-binding protein (LBP) and bactericidal/permeability-increasing protein (BPI) are two structurally related lipid A-binding proteins with divergent functional activities . LBP mediates activation of macrophage and other proinflammatory cells . In contrast, BPI has potent bactericidal and LPS-neutralizing activities . A recombinant fragment of BPI (rBPI23) retains the potent biological activities of the holo protein and may represent a novel therapeutic agent for the treatment of gram-negative infections, sepsis, and endotoxemia . For therapeutic effectiveness in many clinical situations, rBPI23 will have to successfully compete with high serum levels of LBP for binding to endotoxin and gram-negative bacteria . The relative binding affinities of rBPI23 and human recombinant LBP (rLBP) for lipid A and gram-negative bacteria were evaluated . The binding of both proteins to lipid A was specific and saturable with apparent Kds of 2.6 nM for rBPI23 and 58 nM for rLBP . rBPI23 was approximately 75-fold more potent than rLBP in inhibiting the binding of 125I-rLBP to lipid A . The binding affinity of rBPI23 (Kd = 70 nM) for Escherichia coli J5 bacteria was also significantly higher than that of rLBP (Kd = 1,050 nM) . In addition, rBPI23 at 0.2 micrograms/ml was able to inhibit LPS-induced tumor necrosis factor release from monocytes in the presence of 20 micrograms of rLBP per ml . These results demonstrate that rBPI23 binds more avidly to endotoxin than does rLBP and that, even in the presence of a 100-fold weight excess of rLBP, rBPI23 effectively blocks the proinflammatory response of peripheral blood mononuclear cells to endotoxin. Infect Immun, 1994 Apr, 62(4), 1171 - 5 Binding of lysozyme to lipopolysaccharide suppresses tumor necrosis factor production in vivo; Takada K et al.; Endotoxin (lipopolysaccharide {LPS}) released during gram-negative bacterial infection induces varieties of cytokines which directly and/or indirectly cause shock, disseminated intravascular coagulation, and death . We previously showed that lysozyme (LZM) was an LPS-binding protein and inhibited various immunomodulating activities of LPS . In this study, we examined the effect of LZM on the LPS-triggered septic shock model induced by carrageenan treatment and assessed by tumor necrosis factor production . The data presented in this report strongly suggest that LZM-LPS complex formation completely abrogates tumor necrosis factor production and the mortality caused by LPS and that LZM may be useful for the treatment of endotoxin shock. Wei Sheng Wu Xue Bao, 1994 Apr, 34(2), 96 - 9 {A new species of Pseudomonas}; Zou G et al.; A strain No . 9191 isolated from fruit juice mineral drink is gram-negative short-rod . It differs from reported species of Pseudomonas in morphological, physiological and biochemical characteristics . Main characteristics of No . 9191 are single polar flagelled, oxidase and catalase positive, no acid from glucose but producing alkali, nitrate reduced to nitrite, inability to hydrolize gelatin and starch, arginine dihydrolase absent, no growth in presence of 0.85% NaCl . The G+C mol% of DNA is 65.15 . It is named Pseudomonas halosensibilis Zou & Cai nov . sp . by the sensibility property for salt. Inflammation, 1994 Apr, 18(2), 221 - 33 Signal transduction pathways of bacterial lipopolysaccharide-stimulated bovine vascular endothelial cells; Yang Z et al.; Increased procoagulant activity of vascular endothelial cells may be an important component in the pathogenesis of intravascular coagulation associated with gram-negative bacterial diseases . Two bovine endothelial cell (BEC) lines isolated from pulmonary arteries (ENS-2 and ENT-18) were used in this study to investigate procoagulant signal transduction pathways of endotoxin (lipopolysaccharide, LPS)--stimulated BECs . The endothelial cell line ENS-2 was sensitive to LPS as demonstrated by tissue factor (TF) expression, but in contrast, the ENT-18 endothelial cell line was unusually resistant to the effects of LPS . No remarkable quantitative difference in binding of radiolabeled LPS was detected between the two endothelial cell lines . A protein kinase C (PKC) activator (phorbol 12-myristate 13-acetate, PMA) failed to induce TF expression in either cell line at concentrations ranging from 0.05 to 1.00 microM when used as a sole stimulus for the endothelial cells . However, when PMA was used in combination with LPS, PMA enhanced the stimulatory effect of LPS on the endothelial cells . In parallel experiments, PKC inhibitors (H-7 and GF 109203X) interfered with the stimulatory effect of LPS on the cells by decreasing tissue factor expression . We also found that an activator of adenylate cyclase, forskolin, similarly inhibited LPS-induced tissue factor activity . In contrast, protein tyrosine kinase inhibitors (genistein, lavendustin A) had no inhibitory effect on LPS-induced endothelial cell tissue factor expression . Our results collectively suggest that activation of PKC is an important step in stimulation of endothelial cells by LPS, and that LPS and phorbol esters may synergize to produce an enhanced stimulatory effect . Our results also suggest participation of cAMP in controlling LPS-mediated stimulation of endothelia