Gram negative

Eur J Clin Microbiol Infect Dis, 1992 Sep, 11(9), 782 - 8
Patterns of multiple resistance to antibiotics in gram-negative bacteria demonstrated by factor analysis; Leibovici L et al.; Principal component analysis was used to demonstrate the main associations between patterns of resistance to antibiotic drugs in 670 gram-negative bacteria consecutively isolated from blood cultures over a period of two years . Six factors were derived, which accounted for 84% of the total variance of the original matrix . Each factor represented an association between resistance to certain antibiotics as follows: factor 1: aztreonam, third generation cephalosporins and aminoglycosides; factor 2: first and second generation cephalosporins; factor 3: tetracycline and chloramphenicol; factor 4: ampicillin and ureidopenicillins; factor 5: trimethoprim/sulfamethoxazole; factor 6: fluoroquinolones . On two-way analysis of variance the difference in the factor scores was significant between bacteria for all factors except factor 5 . The difference in factor scores between community and hospital acquired strains was significant only for factors 1, 2 and 6 . Only the score of factor 6 showed a clear trend to increase with time during the two-year study period . Patients who were treated with antibiotics prior to bacteremia had higher scores for all factors, the difference being most marked in patients treated with fluoroquinolones . Factor analysis can be used to describe phenotypic associations between resistance to antibiotics, and the factor score used to compare groups of isolates and to demonstrate temporal and other trends.

Clin Investig, 1992 Sep, 70(9), 825 - 42
The inflammatory function of renal glomerular mesangial cells and their interaction with the cellular immune system; Radeke HH et al.; The autoimmune nature of chronic progredient glomerular diseases has been well established . Like in other chronic inflammatory diseases, the active role of organ-borne cells has become increasingly apparent--both for the inflammatory process and for the initiation and perpetuation of the immune reaction . In most forms of glomerulonephritis, intrinsic glomerular mesangial cells are likely candidates to come into intimate contact with immune cells such as monocytes or lymphocytes . On the basis of cell culture studies we would like to integrate the current knowledge available about the responsiveness of mesangial cells to inflammatory agents and the resulting secretory capacity and, moreover, their possible role in sustaining chronic inflammatory injury and autoimmune reactions through a direct interaction with lymphocytes . Apart from being responsive to physiological stimuli such as angiotensin II, glomerular mesangial cells are predominantly activated by agents related to inflammation . This includes exogenous substances such as the components of gram-negative bacteria and an array of highly potent immunological stimuli like antigen-antibody complexes, activated complement, or various cytokines . The transformation of resting mesangial cells to proliferating cells with an accompanying expansion of their secretory profile and responsiveness is due to mediators like platelet-derived growth factor, transforming growth factor, and others . Numerous low-molecular-weight substances (O2-, H2O2, NO, platelet-activating factor, eicosanoids), proteins (proteinases, matrix components, interleukins 1 and 6, colony-stimulating factors, growth factors), and cell-surface molecules released or expressed by mesangial cells participate in the inflammatory process . Among these products interleukin 1 and/or 6, class II major histocompatibility antigen and integrins also support an interaction with the cellular immune system . It has been well documented that mesangial cells induced in vitro by recombinant T-cell lymphokines, such as interferon-gamma, do express MHC II and ICAM-1 and could function as antigen-presenting cells . However, and perhaps more interestingly, our own recent experiments with cocultures of syngeneic mouse lymphocytes and mesangial cells have demonstrated that T-cells are directly activated by cultured mesangial cells, thus resembling a mesangial cell-specific autoimmune reaction . In parallel to clinical studies searching for a mesangial autoantigen these experiments might help to elucidate the mechanisms of initiation and perpetuation of mesangial cell-dependent autoimmune glomerulonephritis.

Z Naturforsch {C}, 1992 Sep-Oct, 47(9-10), 757 - 61
Electrostatic potential barrier in asymmetric planar lipopolysaccharide/phospholipid bilayers probed with the valinomycin-K+ complex; Seydel U et al.; Using the carrier-ion complex valinomycin-K+, current/voltage (I/U) characteristics were registered for planar asymmetric lipid bilayers composed on one side of a phospholipid mixture and on the other side of rough mutant lipopolysaccharide . This system resembles the lipid matrix of the outer membrane of Gram-negative bacteria . The evaluation of the current/voltage curves yielded a highly asymmetric electrical potential barrier . The total potential difference between the phospholipid and the lipopolysaccharide was -85 mV, a result which cannot be explained by contributions of Gouy-Chapman potentials alone . The possible contribution of dipole potentials and influences of headgroup effects are discussed . It is shown that the asymmetry of the I/U-characteristic results from the differences of the surface charge densities of the two monolayers but not from those of the states of order of their hydrocarbon chains.

FEMS Microbiol Immunol, 1992 Sep, 5(1-3), 37 - 43
Activation of Escherichia coli prohemolysin to the membrane-targetted toxin by HlyC-directed ACP-dependent fatty acylation; Hughes C et al.; Hemolysin (HlyA) and related toxins of Escherichia coli and other Gram-negative pathogenic bacteria form membrane pores in cells of the host immune system, causing cell dysfunction and death . An insight into the mechanism by which HlyA is targetted to mammalian cell membranes was achieved by establishing in vitro activation of the non-toxic precursor proHlyA . By this approach we have discovered that conversion of proHlyA to the post-translational active HlyA toxin is determined by fatty acylation of proHlyA in an apparently novel process directed by the HlyC homodimer activator protein, and dependent upon the cellular acyl carrier protein (ACP) . By further exploiting the in vitro activation system it is now possible to obtain direct evidence that HlyC binds to an internal recognition sequence in the proHlyA precursor, in this way providing specificity for the transfer to proHlyA of a fatty acid moiety carried by the ACP . It is possible that the fatty acid modification determines directly the binding of HlyA to mammalian membrane lipids, thus initiating the toxin interaction with the target cells.

Antimicrob Agents Chemother, 1992 Sep, 36(9), 1971 - 7
Studies of the effect of a platelet-activating factor antagonist, CL 184,005, in animal models of gram-negative bacterial sepsis; Torley LW et al.; The effect of CL 184,005, a potent and specific platelet-activating factor antagonist, has been examined in a variety of animal models relevant to gram-negative bacterial sepsis . Pretreatment of mice with CL 184,005 protected them from the lethal effects of platelet-activating factor . When rats or primates rendered hypotensive with endotoxin were treated with CL 184,005, blood pressure was normalized . Pretreatment of rats with CL 184,005 protected them from the gastrointestinal lesions induced by endotoxin . Pretreatment of rats and mice with CL 184,005 protected them from the lethal effects of endotoxin . Plasma tumor necrosis factor levels in endotoxin-treated mice were lower when the mice were pretreated with CL 184,005 . These observations suggest that CL 184,005 may be potentially useful in the treatment of gram-negative bacterial sepsis, and the agent is undergoing clinical evaluation.

Int J Hyperthermia, 1992 Sep-Oct, 8(5), 689 - 99
Lethal interaction between heat and methylene blue in Escherichia coli; Menezes S et al.; Hyperthermia treatment is shown to act synergistically with methylene blue (MB), from the end point of lethality in Gram-negative Escherichia coli bacteria . That this lethality is correlated to the damage produced in DNA by the dye is deduced from the fact that bacteria differing in capacity for repair are almost equally sensitive to heat, but differ considerably in sensitivity to concomitant heat and dye treatment . It is demonstrated that the damage is repairable by the excision-repair system . The role of temperature seems to be that of facilitating the incorporation of the dye, which enables the latter to intercalate into the DNA . Ability of the outer membrane of E . coli AB1157 bacteria to act as a barrier to the penetration of MB remains almost intact up to 46 degrees C, but above this temperature it seems to disrupt abruptly (but reversibly), leading to inactivation of the cells by the dye . Since hyperthermia is in current use for the treatment of cancer, it is suggested that if this synergism also exists in mammalian cells, MB could eventually be used independently of its photodynamic action as an adjuvant in cancer therapy.

J Clin Microbiol, 1992 Sep, 30(9), 2506 - 8
Osteomyelitis and intervertebral discitis caused by Pseudomonas pickettii; Wertheim WA et al.; Pseudomonas pickettii, a nonfermenting, gram-negative rod, is rarely pathogenic . Previous reports of infection with P . pickettii have largely involved direct contamination of supplies presumed to be sterile . We describe a case of vertebral osteomyelitis and intervertebral discitis caused by P . pickettii in a debilitated patient . The aggressive nature of this infection demonstrates that P . pickettii may be a more invasive organism than previously noted, particularly in hosts with weakened immunity secondary to underlying disease.

J Appl Physiol, 1992 Sep, 73(3), 925 - 31
Effects of ibuprofen and pentoxifylline on the cardiovascular response of normal humans to endotoxin; Martich GD et al.; Endotoxin is a major mediator of the life-threatening cardiovascular dysfunction that characterizes Gram-negative sepsis . In animal models of endotoxemia, pretreatment with ibuprofen or pentoxifylline attenuates some of these cardiovascular changes . To evaluate the effects of these agents on the human cardiovascular response to endotoxemia, hemodynamic variables were measured serially in 24 normal subjects who were given intravenous endotoxin . The subjects were randomized to receive oral ibuprofen (n = 9), pentoxifylline (n = 10), or no medication before endotoxin administration (n = 5) . The subjects were volume loaded 3-5 h after endotoxin administration, and hemodynamic measurements were reassessed . Core temperature after endotoxin alone or endotoxin-pentoxifylline approached a maximum at 3 h (greater than or equal to 38.6 degrees C), while the endotoxin-ibuprofen group remained afebrile . At 3 and 5 h, all three groups had significant increases in heart rate, cardiac index, oxygen delivery, and oxygen consumption, while systemic vascular resistance index decreased significantly from baseline . The oxygen extraction ratio remained unchanged . After volume loading, the left ventricular ejection fraction and left ventricular end-diastolic and end-systolic volume indexes did not differ among the groups . The hyperdynamic cardiovascular response to endotoxin in humans occurs in the absence of fever and is not significantly ameliorated by oral cyclooxygenase or phosphodiesterase inhibition.

Int J STD AIDS, 1992 Sep-Oct, 3(5), 355 - 9
Chlamydia trachomatis antigen detection by Chlamydiazyme combined with Chlamydia Blocking Reagent verification; Zeeberg B et al.; Several options exist for the detection of chlamydial infection in a routine laboratory setting . Enzyme immuno assay (EIA) technology offers rapid turn around of results and is less technically demanding than chlamydial cell culture . In addition, recently introduced EIA confirmatory reagents have the potential to improve the accuracy of EIA detection . We have evaluated one such confirmatory reagent (Chlamydia Blocking Reagent, Abbott Laboratories) to determine the accuracy of the Chlamydiazyme EIA with special regard to interpretation of low absorbance values . An initial series of 192 male urethral specimens showed that use of a lowered cut off level (absorbance value 0.05) compared with that recommended by the manufacturer increased sensitivity of the EIA from 0.73 to 0.83, thus motivating studies on this interpretative modification . Of 1101 EIA reactive specimens, 65% were determined to be chlamydia positive by the Chlamydia Blocking Reagent . The proportion of female cervical specimens that did not confirm positive was elevated compared with male urethral specimens, 43% vs . 5.7% respectively . In samples yielding absorbance from the recommended cut off level to 0.05 (approximately 50% below), the corresponding figures were 78% and 14% respectively . In 85 selected EIA reactive samples, examination by a direct immunofluorescence staining assay (DFA) (MicroTrak, Syva Inc.) revealed elementary bodies in 85% of 67 blocking test positive and in 24% of 18 blocking test negative samples . The possibility that Gram-negative bacteria were responsible for unconfirmed EIA reactive specimens was investigated using bacterial suspensions . While EIA reactivity was noted with several strains for Gram-negative bacteria, both the blocking reagent and DFA correctly verified the absence of chlamydial antigen.(ABSTRACT TRUNCATED AT 250 WORDS)

Cesk Epidemiol Mikrobiol Imunol, 1992 Sep, 42(3), 120 - 5
{A program for prevention of pseudomonas infections and immunoprophylaxis with PSAEVA vaccine}; Kahlich R et al.; The authors present three main trends of prevention of Gram-negative opportunistic infections investigated at the Cardiosurgical Clinic of the Institute of Clinical and Experimental Medicine . An important part is played by active and systematic surveillance with early signalling and analysis of new infections with subsequent aimed interventions against concrete sources of infection and mechanisms of transmission . It is necessary to think more of activation of endogenous infection the specific features of which make repression difficult and motivate investigations of preventive approaches . Rational antibiotic policy has so far not appreciated the danger of selection of endogenous and exogenous agents in standard situations where antibiotic therapy lacks an aimed and restrained approach . Immunoprophylaxis with the PSAEVA vaccine in the first preliminary controlled clinical investigation was characterized by the absence of nosocomial epidemics and septicaemia and a significant (fivefold) reduction of the incidence of non-pyocyanic Gram-negative bacteriaemia . The success of vaccination seems to be related to the anti-colonization and anti-invasive mechanism of specific and unspecific nature.

Cesk Epidemiol Mikrobiol Imunol, 1992 Sep, 42(3), 102 - 5
{New findings in emergency care and resuscitation in patients at risk for endotoxic shock}; Drabkova J; Endotoxin shock is not only the reflexion of Gram-negative focal infection but also the consequence of dysfunction of the intestinal mucous barrier and a decline of the detoxication capacity, in particular of the hepatic mesenchymal phagocytic system during a critical state . Cytokines and the primary LPS complex and its lipid A resp . are of basic importance . They start the release of a large amount of TNF alpha, IL-1, IL-6, IL-8 and other cascades . Acute shock is controlled nowadays more frequently than in the past, however, there is a high risk of a very adverse reaction of remote organs, which is very adverse from the prognostic aspect . A series of laboratory markers has a greater validity than the clinical picture alone . For screening derived markers are used not primary markers . Despite this they provide adequate information . Prophylaxis and treatment include selective bacterial decontamination, or active or passive immunization (PSAEVA, hyperimmune sera), minidoses of dopamine in a continuous infusion, early enteral nutritional intervention, in particular enteral nutrition containing glutamine . Monoclonal and polyclonal antibodies against the LPS complex and cytokines are tested, blocking their receptors or possibly early plasmapheresis . Permanent pillars of therapeutic tactics are still a radical and early elimination of possible infectious foci and targeted administration of antibiotics and maintenance of the perfusion pressure and adequate oxygenation.

Ann R Coll Surg Engl, 1992 Sep, 74(5), 329 - 34
Prevention of bacterial infection and sepsis in acute severe pancreatitis; McClelland P et al.; Between 1984 and 1986 six patients with acute respiratory failure (requiring ventilation for at least 3 days) complicating acute pancreatitis were managed on the intensive care unit (median ventilation period 6 days; range 3-41 days) . Between 1987 and 1989 nine similar patients were managed (median ventilation period 35 days, range 4-69 days), and a regimen of enteral tobramycin, polymyxin and amphotericin to selectively decontaminate the digestive tract (SDD) was introduced . Five of six patients treated before 1987 had serious infections (three Gram-negative, one fungal), compared with only one of nine patients treated with SDD (P < 0.05) . Clinical signs of sepsis were evident for 62% of the pre-SDD period, compared with 39% of the period during SDD therapy (P < 0.001) . Systemic antibiotic prescribing was reduced in the SDD group; however, mortality remained unaffected with only two patients surviving pre-SDD and three during SDD treatment . SDD reduces infection rates and sepsis in patients with acute pancreatitis and may help to improve the prognosis of this life-threatening condition.

Ugeskr Laeger, 1992 Aug 31, 154(36), 2437 - 8
{Meningitis caused by Capnocytophaga carnimorsus with development of erythema nodosum}; Storgaard M; A case of meningitis in a 42-year-old previously healthy man caused by the gram-negative rod Capnocytophaga carnimorsus is presented . The patient had erythema nodosum . Bacteremia was possibly induced during dental treatment.

Nature, 1992 Aug 27, 358(6389), 727 - 33
Crystal structures explain functional properties of two E . coli porins; Cowan SW et al.; Porins form aqueous channels that aid the diffusion of small hydrophilic molecules across the outer membrane of Gram-negative bacteria . The crystal structures of matrix porin and phosphoporin both reveal trimers of identical subunits, each subunit consisting of a 16-stranded anti-parallel beta-barrel containing a pore . A long loop inside the barrel contributes to a constriction of the channel where the charge distribution affects ion selectivity . The structures explain at the molecular level functional characteristics and their alterations by known mutations.

J Bacteriol, 1992 Aug, 174(16), 5450 - 3
Identification of a putative eukaryotic-like protein kinase family in the developmental bacterium Myxococcus xanthus; Zhang W et al.; Myxococcus xanthus is a gram-negative bacterium which, upon starvation, undergoes a spectacular developmental cycle culminating in the formation of spore-filled fruiting bodies . We recently characterized a protein serine-threonine kinase (Pkn1) that is required for normal development (J . Munoz-Dorado, S . Inouye, and M . Inouye, Cell 67:995-1006, 1991) . pkn1 was cloned by polymerase chain reaction amplification with primers designed from conserved sequences in eukaryotic protein kinases . In this study, a fragment of the pkn1 gene and an oligonucleotide corresponding to another highly conserved region were employed as probes for Southern blot analyses, which indicated that there are at least 26 putative kinase genes in M . xanthus . Most of the putative kinase genes were cloned, and complete or partial sequencing of eight clones revealed that they indeed contained highly conserved sequences present in eukaryotic kinases . These results suggest that complex kinase cascades similar to those described for eukaryotes might be involved in regulation of the M . xanthus life cycle.

J Bacteriol, 1992 Aug, 174(16), 5237 - 43
Identification of amino acid substitutions that alter the substrate specificity of TEM-1 beta-lactamase; Palzkill T et al.; TEM-1 beta-lactamase is the most prevalent plasmid-mediated beta-lactamase in gram-negative bacteria . Recently, TEM beta-lactamase variants with amino acid substitutions in the active-site pocket of the enzyme have been identified in natural isolates with increased resistance to extended-spectrum cephalosporins . To identify other amino acid substitutions that alter the activity of TEM-1 towards extended-spectrum cephalosporins, we probed regions around the active-site pocket by random-replacement mutagenesis . This mutagenesis technique involves randomizing the DNA sequence of three to six codons in the blaTEM-1 gene to form a library containing all or nearly all of the possible substitutions for the region randomized . In total, 20 different residue positions that had been randomized were screened for amino acid substitutions that increased enzyme activity towards the extended-spectrum cephalosporin cefotaxime . Substitutions at positions 104, 168, and 238 in the TEM-1 beta-lactamase that resulted in increased enzyme activity towards extended-spectrum cephalosporins were found . In addition, small deletions in the loop containing residues 166 to 170 drastically altered the substrate specificity of the enzyme by increasing activity towards extended-spectrum cephalosporins while virtually eliminating activity towards ampicillin.

Chest, 1992 Aug, 102(2), 613 - 7
Potential treatment of sepsis syndrome with cytokine-specific agents; Christman JW; Blocking the effects of cytokines is a potential new therapeutic avenue for the treatment of Gram-negative sepsis . Three classes of agents are currently being evaluated: antibodies, circulating inhibitors, and receptor antagonists . Data in the current literature support the consideration of these agents as potential therapeutic agents in Gram-negative sepsis . The clinical utility of these agents is contingent on the results of well-designed, prospective, randomized, placebo-controlled clinical trials in well-defined clinical populations . These trials will require the cooperation of clinical and basic scientists . At this time, preliminary and early clinical trials are in progress utilizing IL-1 and TNF-alpha circulating inhibitors, IL-1 receptor antagonists, and monoclonal antibodies to TNF-alpha and the TNF-alpha receptor.

J Infect Dis, 1992 Aug, 166(2), 331 - 5
Role of interferon-gamma in experimental gram-negative sepsis; Silva AT et al.; To study the role of interferon-gamma (IFN-gamma) in gram-negative shock, mortality was compared in mice receiving either a monoclonal antibody to IFN-gamma (H22) or an irrelevant monoclonal antibody (L2-3D9) before or after an LD90 dose of Escherichia coli O111:B4 . H22 given either 1 h before or 0.5 h after bacterial challenge protected mice from death (mortality at 48 h, 28% vs . 83%, P less than .001) . Serum tumor necrosis factor-alpha (TNF alpha) levels and bacterial counts in blood and organs (liver, spleen, heart, and brain) were similar in H22-treated animals and controls . The peak serum TNF alpha levels were 95.7 +/- 16.4 ng/mL and 80.7 +/- 14.9 ng/mL in the H22 and control groups, respectively . These results indicate that IFN-gamma plays a significant role in the pathogenesis of gram-negative sepsis.

J Virol, 1992 Aug, 66(8), 4720 - 31
Deletion analysis of two tandemly arranged virulence genes in myxoma virus, M11L and myxoma growth factor; Opgenorth A et al.; Myxoma virus (MYX) is a leporipoxvirus of rabbits that induces a lethal syndrome characterized by disseminated tumorlike lesions, generalized immunosuppression, and secondary gram-negative bacterial infection . A MYX deletion mutant (vMYX-GF- delta M11L) was constructed to remove the entire myxoma growth factor (MGF) coding sequence and that for the C-terminal five amino acids of the partially overlapping upstream gene, M11L . Unexpectedly, this deletion completely abrogates the capacity of MYX to cause the characteristic disease symptoms of myxomatosis . Upon inoculation of rabbits with vMYX-GF- delta M11L, recipient animals developed only a benign, localized nodule reminiscent of a Shope fibroma virus-induced tumor in which a single primary lesion appeared at the site of injection and then completely regressed within 14 days, leaving the animals resistant to challenge with wild-type MYX . No evidence of the purulent conjunctivitis and rhinitis that always accompany wild-type MYX infection was observed . To ascertain whether the attenuation observed in vMYX-GF- delta M11L was due to a combined effect of the MGF deletion and alteration of the upstream M11L gene, two additional MYX recombinants were constructed: an MGF- virus (vMYX-GF-) containing an intact M11L gene and an M11L- virus (vMYX-M11L-) containing an intact MGF gene . Infection with vMYX-GF- resulted in moderated symptoms of myxomatosis, but all clinical stages of the disease were still detectable . In contrast, disruption of M11L alone dramatically reduced the virus virulence, resulting in a nonlethal syndrome whose clinical course was nevertheless distinct from that of vMYX-GF- delta M11L . Upon inoculation with vMYX-M11L-, rabbits developed primary and secondary tumors which were larger and more circumscribed than those of wild-type MYX recipients . Whereas wild-type MYX infection always includes severe, purulent conjunctivitis and rhinitis, vMYX-M11L- recipients remained healthy and displayed only minimal signs of respiratory distress . By about 30 days after infection, the tumors induced by vMYX-M11L- had completely regressed and these animals were immune to challenge with wild-type MYX . Histological analysis indicated that tumors induced by vMYX-M11L- are much more heavily infiltrated with macrophages and heterophils and that the sites of viral replication are more edematous and necrotic than those of wild-type infection, suggesting that the host was able to mount a more vigorous inflammatory response to vMYX-M11L- infection.(ABSTRACT TRUNCATED AT 400 WORDS)

Dan Med Bull, 1992 Aug, 39(4), 304 - 20
The molecular biology and diagnostics of Chlamydia trachomatis; Birkelund S; The rapid development of biotechnological methods provides the potential of dissecting the molecular structure of microorganisms . In this review the molecular biology of chlamydia is described . The genus Chlamydia contains three species C . trachomatis, C . psittaci, and C . pneumonia which all are important human pathogens . Chlamydia is obligate intracellular bacteria with a unique biphasic life cycle . The extracellularly chlamydial elementary bodies (EB) are small, metabolic inactive, infectious particles with a tight outer cell membrane . After internalization into host cells the chlamydial structure changes, they transform to reticulated bodies (RB) which become larger, metabolically active, and start to replicate . Fourtysix hrs post infection RB reorganizes to EB followed by burst of the inclusion . The structure of the EB outer membrane differs from the membrane of gram-negative bacteria since it is highly cross-linked by S-S bridges . There are, however, also similarities to gram-negative cell walls . The chlamydial major outer membrane protein, Omp1, forms pores and is closely associated with lipopolysaccharide, LPS . LPS, however, is more loosely associated with Omp1 than in other gram negative bacteria since incubation of EB with antibodies against LPS will liberate it from the chlamydial surface . Therefore the surface localized LPS may be important for chlamydial survival . OMP1 varies between the different serovar of C . trachomatis . Several very conserved regions are separated by variable domains . The variable domains are very antigenic and are localized at the surface of EB . After chlamydial internalization into the host cell transition to RB starts . Some of the early proteins are DnaK-like and groEL-like heat-shock proteins . The chlamydial DnaK-like protein is very antigenic . Patient serum samples will recognize the chlamydial DnaK-like protein . From the determined DNA sequence the amino acid sequence was determined . It was 57% homologous to the Eschrichia coli DnaK protein . Also the GroEL-like protein is antigenic and very conserved . Factors of importance for pathogenicity of chlamydia have not yet been found . The adhesin(s) is unknown, and no factor of importance for the inhibition of fusion between phagosome and host cell lysosomes has been described . A protein similar to the mip gene product of Legionella pneumofila may be a possible candidate for a pathogenicity factor . Diagnosis of C . trachomatis infections has been done by chlamydia cultivation in tissue culture cells, by immunofluorescence and by ELISA . A new method based on the polymerase chain reaction (PCR) has been developed . As primers sequences from the common plasmid were used . This method has high sensitivity and specificity and does not require live chlamydia.(ABSTRACT TRUNCATED AT 400 WORDS)

APMIS, 1992 Aug, 100(8), 747 - 51
IgG subclass antibodies against Helicobacter pylori heat-stable antigens in normal persons and in dyspeptic patients; Andersen LP et al.; Measurement of serum IgG antibodies against Helicobacter pylori seems to be useful in the diagnosis of H . pylori infections . IgG subclass antibodies against H . pylori have, however, not been investigated thoroughly . In this study IgG1, IgG2, IgG3 and IgG4 antibody levels against H . pylori were measured using an ELISA technique in 187 normal adult persons and in 174 patients with dyspeptic symptoms, of whom 99 patients were H . pylori positive . None of the IgG subclass antibody levels were better than the total IgG level for the diagnosis of H . pylori infection . The discrimination between H . pylori-positive and H . pylori-negative patients was better with IgG1, IgG2 and IgG4 antibody levels than with IgG3 antibody level . IgG2 was the IgG subclass antibody that mainly contributed to the age-dependent increase in the IgG antibody level . This sustains the suspicion that cross-reactions between lipopolysaccharide (LPS) from H . pylori and LPS from other Gram-negative bacteria may occur.

Clin Invest Med, 1992 Aug, 15(4), 318 - 30
A new bioactive molecule for improving vascular graft patency: exploratory trials in dogs; Akoum A et al.; Myxalin is a new bioactive molecule that we have isolated from the culture medium of Myxococcus xanthus, a non-pathogenic Gram negative bacterium . This glycopeptide possesses an antithrombotic effect in vivo and has been shown to promote human endothelial cell growth in vitro . With the object of exploring its ability to improve vascular graft healing and patency, myxalin was immobilized on 6 mm diameter knitted polyester prostheses using gelatin as a carrier, and the prosthesis was then implanted as an infrarenal abdominal arterial substitute in dogs for a period of 2 weeks . Two additional series of implantations were conducted for control purposes: one with gelatin-coated prostheses without myxalin, the other following normal preclotting of the polyester grafts . In order to select adequate sterilization conditions which can preserve the biological activity of myxalin, the prostheses were sterilized according to 3 different sterilization processes (gamma radiation and ethylene oxide either at 63 degrees C or 37 degrees C) . At the sacrifice, all grafts were patent . The myxalin treated prostheses exhibited improved blood compatibility in terms of fewer thrombotic deposits and significant inhibition of platelet and fibrinogen uptake on their luminal surfaces . In addition, the development of a thin collagenous internal capsule with endothelial cells secreting high levels of prostacyclin was observed at both anastomoses of the myxalin-treated grafts sterilized by gamma radiation.

Appl Environ Microbiol, 1992 Aug, 58(8), 2543 - 51
Comparison of bioaerosol sampling methods in barns housing swine; Thorne PS et al.; The air in livestock buildings contains bioaerosol levels that are sufficiently high to cause adverse health effects in animals and workers . These bioaerosols are complex mixtures of live and dead microorganisms and their products as well as other aeroallergens . The effectiveness of sampling methods used for quantifying the very high concentrations of microorganisms in these environments has not been well studied . To facilitate an accurate assessment of respiratory hazards from viable organisms in agricultural environments, three bioaerosol sampling methods were investigated: the Andersen microbial sampler method (AMS), the all-glass impinger method (AGI), and the Nuclepore filtration-elution method (NFE) . These methods were studied in a parallel fashion in 24 swine confinement buildings . Measurements were taken in two seasons with three types of culture media in duplicate to assess total bacteria, gram-negative enteric bacteria, and total fungi . Methods were analyzed for the proportion of samples yielding data within the limits of detection, intraclass reliability, and correlation between methods . For sampling viable bacteria, the AMS had a poor data yield because of overloading and demonstrated weak correlation with the AGI . Conversely, the AGI and NFE gave sufficient numbers of valid data points (90%), yielded high intraclass reliabilities (alpha greater than or equal to 0.92), and were highly correlated with each other (r = 0.86) . The AGI and the NFE were suitable methods for assessing bacteria in this environment, but the AMS was not . The AMS was the only method that consistently recovered enteric bacteria (73% data yield) . For sampling fungi, the AGI and AMS both yielded sufficient data and all three methods demonstrated high intraclass reliability.(ABSTRACT TRUNCATED AT 250 WORDS)

Am J Pathol, 1992 Aug, 141(2), 307 - 17
In vivo effects of endotoxin on intraepithelial mucosubstances in rat pulmonary airways . Quantitative histochemistry; Harkema JR et al.; Bacteria-induced bronchopneumonias are often characterized by an influx of neutrophils and excess mucus in pulmonary airways . This study determined how endotoxin, a component of gram-negative bacteria and a potent inflammatory agent, affects the ultrastructure of the mucociliary apparatus and the amount of stored intraepithelial mucosubstances in the main axial airways within the lung . Rats were intranasally instilled, once a day for 3 days, with endotoxin or saline (controls) . Animals were sacrificed 1, 2, or 7 days after the last instillation . Microdissected intrapulmonary axial airways (generations 8-11) from the right caudal lobes of infusion-fixed lungs were processed for light and electron microscopy . Morphometric techniques were used to determine the volume densities (Vs) of histochemically stained intraepithelial mucosubstances and numerical densities of airway epithelial cells . There were marked increases, compared with controls, in the amount of intraepithelial mucosubstances in the intrapulmonary axial airways at generations 8 and 11 in the right caudal lobes from endotoxin-instilled rats sacrificed 1, 2, and 7 days after the last instillation . There were significantly greater numbers of surface epithelial cells per length of basal lamina (i.e., hyperplasia) in endotoxin-exposed airways compared with airways from controls . This endotoxin-induced hyperplasia was due primarily to an increase in the number of mucus-secretory cells, which in endotoxin-exposed epithelium were columnar and contained numerous, large confluent, electronlucent, secretory granules composed of acidic and neutral glycoproteins . In contrast, secretory cells in airway epithelium from controls were cuboidal and contained small discrete, electron-dense, granules composed of only neutral glycoproteins . The numbers of ciliated cells and basal cells were similar in both control and endotoxin-exposed epithelium . Only endotoxin-exposed epithelium, however, contained atypical epithelial cells with numerous basal bodies, few cilia, and few apical secretory granules . These results indicate that repeated airway instillations of endotoxin induce an increase in the amount of intraepithelial mucosubstances, secretory cell hyperplasia, and excess luminal mucus in pulmonary airways . Therefore, endotoxin released from gram-negative bacteria may be partially responsible for the structural alterations, in the airway surface epithelium, which result in the excess luminal mucus observed in bacteria-induced bronchopneumonias.

J Trop Med Hyg, 1992 Aug, 95(4), 271 - 5
Detection of Brucella melitensis and Brucella abortus by DNA amplification; Baily GG et al.; Suitable reaction conditions and oligonucleotide primers were sought for the detection of Brucella melitensis and Brucella abortus by the polymerase chain reaction . Primers were chosen from within the coding sequence of a gene encoding a 31 kDa B . abortus antigen . The test was shown to be sensitive, and specificity was demonstrated using DNA derived from a panel of Gram-negative pathogens . There was no detectable difference between B . melitensis and B . abortus in the sensitivity of the reaction or in the size of the amplification product . The technique should be applicable in the diagnosis of brucellosis.

Am Rev Respir Dis, 1992 Aug, 146(2), 352 - 7
Inflammatory response to acute inhalation of endotoxin in asthmatic patients; Michel O et al.; Inhalation of 20 micrograms endotoxins (from the membrane of Gram-negative bacteria) has been reported to induce a bronchial obstructive response in asthmatic subjects . The aim of the present study was to evaluate in asthmatic patients the possibility of an inflammatory response to inhaled endotoxins . Eight patients with mild asthma were submitted to bronchial challenge tests, in a single-blind trial, on Day 1 with control solution and on Day 7 with 20 micrograms endotoxin of Escherichia coli (026:B6) . Local inflammatory response was indirectly evaluated by the degree of bronchial hyperresponsiveness (BHR) expressed as PD20 FEV1 histamine (the dose of histamine inducing a 20% decrease in FEV1) at 0, 6, 24, and 48 h and 7 days . Systemic inflammation was investigated by sequential blood determinations of total (and differential) white cells, complement anaphylatoxin C5a, interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha), and C-reactive protein (CRP) . A significant (p < 0.01) bronchial obstructive response was demonstrable 45 min after lipopolysaccharide (LPS) inhalation, lasting 5 h . Comparing the level of BHR after control inhalation, a significant (p < 0.05) increase in BHR was shown 6 h after LPS, partially normalized at 24 and 48 h . A short peak in TNF-alpha at 60 min (p < 0.05) and an increase in total white blood cells (p < 0.01) and neutrophil polymorphonuclear neutrophils at 360 min (p < 0.05) and of CRP at 24 and 48 h (p < 0.05 and p < 0.01) were significant . The other blood parameters did not change significantly.(ABSTRACT TRUNCATED AT 250 WORDS)

J Am Osteopath Assoc, 1992 Aug, 92(8), 1017 - 20
Update on monoclonal antibody therapy in the gram-negative sepsis syndrome; Hecker RB; Two multicenter controlled clinical trials of genetically engineered monoclonal antibodies directed against endotoxin, a potent mediator of inflammation in the gram-negative sepsis syndrome, were recently reported in the medical literature . One of these antiendotoxin antibodies was derived from a murine (mouse) source, and the other antibody was derived mainly from a human source (nebacumab {negative bacteria human monoclonal antibody}) . This article reviews recent literature concerning the use of these agents in the treatment of gram-negative sepsis syndrome . It also projects economic assessment data regarding the use of nebacumab in the United States.

Z Gastroenterol, 1992 Aug, 30(8), 543 - 52
{Spontaneous bacterial peritonitis}; Press AG et al.; Patients with liver cirrhosis and ascites suffer from spontaneous bacterial peritonitis (SBP) in up to 25% . The typical clinical signs are abdominal pain with tenderness and fever . 30% have no signs of peritonitis . Then clinical worsening, encephalopathy, rising serum creatinine levels, and therapy resistant ascites may be the only clinical features . SBP must be differentiated from bacterascites and culture negative neutrocytic ascites by the polymorphonuclear neutrophil (PMN) count in the ascites and the presence of positive culture results, which has prognostic implications . Gram negative rods from the colon play an important etiological role in SBP . Gastrointestinal bleeding, lack of serum complement, a low ascites protein and the extent of intrahepatic shunts predispose to SBP . Then, prophylaxis with the comparable drugs neomycin and norfloxacin is indicated . Coexisting encephalopathy has to be treated by the therefore effective neomycin . Otherwise, norfloxacin is the drug of choice because of better acceptance and lower costs . Chemical parameters of the ascites (pH value less than 7.4; LDH and lactate greater than serum levels; glucose less than 50 mg%) help to assess the severity of peritonitis . The course of ascitic PMN under therapy and the time of persisting positive cultures can discriminate SBP from secondary peritonitis . Antibiotics of choice are amoxicillin-clavulanic acid and cefotaxime . Short course therapy (5 days) is a effective as long course therapy (10 days) . Today SBP is no more life-threatening because diagnosis, prophylaxis and therapy have improved . However, complication rate of patients with liver cirrhosis and ascites has not changed.

Childs Nerv Syst, 1992 Aug, 8(5), 279 - 80
Brain abscess in infants; Krajewski R et al.; Brain abscesses are rare in infants and their clinical presentation is specific for this age group . Seven cases of brain abscess in infants aged 2-11 months are reported . The underlying cause was meningitis in four, sepsis in two, and unknown in one . Gram-negative organisms were cultured in 6 patients . The abscess size was 5 cm or more in five cases; in four there were multiple lesions . Two abscesses were aspirated and irrigated; four particularly large lesions were drained and repeatedly aspirated and irrigated . One craniotomy was done . There were two deaths, one in the postoperative period and the other 6 months after discharge . Follow-up information is available for four children, showing a good result in only one of them . Formation of an abscess should be diagnosed early, and close ultrasound monitoring or CT scanning in infants with bacterial meningitis and sepsis is essential . The prognosis in cases in which large/multiple abscesses develop is poor.

Biokhimiia, 1992 Aug, 57(8), 1196 - 205
{Selective solubilization and biochemical analysis of R . prowazekii outer membrane proteins}; Emel'ianov VV; Solubilization of proteins from total membranes (a mixture of cytoplasmic and outer membranes) of Rickettsia prowazekii, a typical gram-negative bacterium, was studied using three different detergents . It was shown that isolated outer membranes and sarkosyl-insoluble material contain major polypeptides of 134, 31, 29.5 and 25 kDa as well as minor polypeptides of 78, 60, 42, and 17 kDa, while the total membranes--the same plus a great number of additional minor proteins . The material solubilized by octyl glucoside in the presence of MgCl2 contains exclusively major proteins (134, 31, 29.5, and 25 kDa) . No differential solubilization takes place upon membrane treatment with octyl glucoside in the absence of Mg2+ or with Triton X-100 . Rickettsial proteins are insensitive to trypsin in both whole cells and total membranes, unless the latter are presolubilized with octyl glucoside . Proteinase K degrades all of the total membrane proteins but only the 134 kDa polypeptide of whole cells . Upon immunoblotting predominantly the major outer membrane proteins (134, 31, and 20.5 kDa) and, to a lesser extent, the minor proteins (60, 42, and 17 kDa) interact with human convalescent serum.

Br J Haematol, 1992 Aug, 81(4), 480 - 8
Modulation of tissue factor on human monocytes by cisplatin and adriamycin; Walsh J et al.; Coagulation disorders have been associated with the use of chemotherapeutic drugs . Pharmacological doses of cisplatin and adriamycin directly induced low levels of procoagulant on normal human blood monocytes and on a human myelomonocytic cell line, RC2a . Activity was maximal after 24 h and was not due to cell lysis as increasing drug doses which decreased cell viability were less effective . Procoagulant induction was markedly enhanced in the presence of bacterial lipopolysaccharide (LPS), with as little as 10-100 pg/ml LPS potentiating the cisplatin response by 2-5-fold and more than doubling the adriamycin response . Greater than 90% of the procoagulant activity was membrane-bound tissue factor as indicated by the factor VII-dependent generation of factor Xa by viable cells and by the neutralization of this activity by a monoclonal antibody to tissue factor . Tissue factor antigen was measured simultaneously by immunohistochemical staining and by cell ELISA . Blood monocytes activated with LPS expressed high levels of tissue factor antigen; by contrast, adriamycin and cisplatin did not appear to induce antigen expression, but to enhance the specific activity of that already present . Results suggest that membrane alterations which occur following treatment with DNA/RNA intercalating drugs, may result in a highly active form of monocyte/macrophage tissue factor which may contribute to the complications caused by activated coagulation . Secondary Gram-negative infection or cytokines released by an active immune response to a tumour may contribute to the procoagulant potential of these cytotoxic drugs.

Surgery, 1992 Aug, 112(2), 188 - 93; discussion 193-4
Recombinant interleukin-1 receptor antagonist (IL-1ra): effective therapy against gram-negative sepsis in rats; Alexander HR et al.; BACKGROUND . Morbidity and mortality from bacterial sepsis remain high despite aggressive diagnostic and therapeutic intervention . Interleukin-1 has been implicated as mediator of the lethal effects of endotoxemia or bacterial sepsis . The current experiments were designed to evaluate the therapeutic efficacy of a human recombinant interleukin-1 receptor antagonist (IL-1ra) against polymicrobial gram-negative septicemia in rats . METHODS . Male rats underwent placement of indwelling carotid arterial and superior vena caval catheters followed by cecal ligation and puncture (CLP) . After 3 hours rats received either IL-1ra (10 mg/kg intravenous bolus followed by 5 mg/kg/hr) or an equal volume of vehicle intravenously for 24 hours . Heart rate, respirations, mean arterial blood pressure, and temperature were recorded at frequent intervals, and survival was assessed for 30 hours after CLP . RESULTS . There were no differences in vital signs between groups at baseline or before treatment, and all animals appeared ill with huddled posture, piloerection, and hyperventilation . Twenty-four hours after CLP, IL-1ra significantly ameliorated bradycardia (p = 0.01), hypothermia (p = 0.001), and hypotension (p = 0.05), and 30-hour survival was significantly improved (71% vs 20%, p less than 0.05) . CONCLUSIONS . IL-1ra lessens the acute hemodynamic, hypothermic, and mortal effects of gram-negative sepsis induced by CLP in rats . These data suggest that IL-1 receptor blockade may be an important new treatment strategy against overwhelming bacterial sepsis.

J Immunol, 1992 Aug 1, 149(3), 918 - 24
Granulocyte colony-stimulating factor treatment protects rodents against lipopolysaccharide-induced toxicity via suppression of systemic tumor necrosis factor-alpha; Gorgen I et al.; Pretreatment with recombinant human granulocyte CSF (G-CSF) protected mice in two different models of septic shock . Intravenous injection of 250 micrograms/kg G-CSF to mice prevented lethality induced by 5 mg/kg LPS . Injection of 50 micrograms/kg G-CSF protected galactosamine-sensitized mice against LPS-induced hepatitis . In either case, this protection was accompanied by a suppression of LPS-induced serum TNF activity . In contrast, when galactosamine-sensitized mice were pretreated with 50 micrograms/kg murine recombinant granulocyte/macrophage CSF instead of G-CSF and subsequently challenged with LPS, serum TNF activity was significantly enhanced and mortality was increased . The suppressive effect of G-CSF on LPS-induced TNF production was also demonstrated in rats . In vivo, no TNF was detectable in the blood of LPS-treated rats, which had been pretreated with G-CSF . Ex vivo, alveolar macrophages, bone marrow macrophages, Kupffer cells, or peritoneal macrophages prepared from G-CSF-treated rats produced significantly less TNF upon stimulation with LPS than corresponding populations from control rats . However, when these macrophage populations were incubated with G-CSF in vitro, LPS-induced TNF production was unaffected . These data suggest that the G-CSF-mediated suppression of TNF production is not a direct effect of G-CSF on macrophages . To examine whether, independent of the protection against LPS, G-CSF treatment still activated neutrophils, it was demonstrated that granulocytes from G-CSF-treated rats were primed for PMA-induced oxidative burst and for ionophore/arachidonic acid-stimulated lipoxygenase product formation . The experiments of this study support the notion that G-CSF is a negative feedback signal for macrophage-derived TNF-alpha production during Gram-negative sepsis.

N Engl J Med, 1992 Jul 23, 327(4), 234 - 40
Prophylactic intravenous administration of standard immune globulin as compared with core-lipopolysaccharide immune globulin in patients at high risk of postsurgical infection . The Intravenous Immunoglobulin Collaborative Study Group.
{Necrobacillosis}
Pedersen G, Hagelskjaer LH.

Medicinsk afdeling B (haematologi) og klinisk mikrobiologisk afdeling, Aalborg Sygehus SydNecrobacillosis is an infection caused by the anaerobic Gram-negative rod Fusobacterium necrophorum . The infection is most common in previously healthy young adults and is characterised by sore throat followed by rigors, septicaemia and the formation of metastatic abscesses, often in the lungs . The infection has a certain mortality, which is reduced when early and sufficient treatment is administered . The treatment is a prolonged course of penicillin and/or metronidazole . Two case histories and a review of the literature are presented . The purpose of this article is to make the clinician aware of the syndrome, which is so characteristic that the diagnosis can be made from the clinical picture alone before bacteriological verification.

JPEN J Parenter Enteral Nutr, 1992 Jul-Aug, 16(4), 353 - 8
Adaptation to a fish oil diet before inducing sepsis in rats prevents fatty infiltration of the liver; Lanza-Jacoby S et al.; Hypertriglyceridemia and fatty liver are common lipid abnormalities associated with Gram-negative sepsis . Fish oils have been shown to have beneficial effects in reducing plasma triglycerides (TG) . This study was designed to investigate whether fish oils would prevent the elevation of plasma TG and the accumulation of liver lipids during sepsis . One group of rats was fed a 10% menhaden oil diet and the other group was fed a 10% corn oil diet for 14 days . On the 14th day, sepsis was induced by injecting the rats with 8 x 10(7) live Escherichia coli colonies/100 g of body weight and the rats were fasted for 22 hours . The liver composition of total lipids and TG in the septic rats prefed the fish oil was lower than in the septic rats prefed the corn oil . In the rats adapted to the corn oil diet, lipids accumulated in the livers of the septic rats in comparison with the control rats . Hepatocytes isolated from the septic rats adapted to the corn oil diet showed an increased esterification of {1-14C}palmitate into TG and phospholipids than hepatocytes from the control rats . Feeding the fish oil diet instead of the corn oil diet before inducing sepsis reduced TG, cholesterol, and phospholipid synthesis by 58%, 79%, and 71%, respectively . The rise in TG synthesis in the septic rats prefed the corn oil diet was associated with an 89% increase in the activity of phosphatidate phosphohydrolase . There was no significant difference in the activities of glycerol-3-phosphate acyltransferase and phosphatidate phosphohydrolase between control and septic rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Appl Environ Microbiol, 1992 Jul, 58(7), 2245 - 9
Numerical taxonomy of gram-negative, nonmotile, nonfermentative bacteria isolated during chilled storage of lamb carcasses; Prieto M et al.; A numerical taxonomic study using 75 characters was performed with 132 strains of gram-negative, nonmotile, nonfermentative bacteria selected on the basis of lack of motility and Gram reaction among 1,200 cultures isolated during aerobic storage of lamb carcasses . At the 80% similarity level (SSM), eight clusters were formed . Strains in clusters 1 to 6 could be identified as members of the family Moraxellaceae and, more specifically, as members of the Psychrobacter-{Moraxella} phenylpyruvica subgroup . Of these strains, clusters 1 and 2 (88 strains) were identified as {Moraxella} phenylpyruvica and cluster 3 (15 strains) was identified as Psychrobacter immobilis . Clusters 4, 5, and 6 were not identifiable with any species . Clusters 7 and 8 consisted of 14 strains considered nonmotile variants of Pseudomonas fragi . The highest separation indices corresponded to acid production from certain carbohydrates (melibiose, L-arabinose, and cellobiose) . Although strains of Psychrobacter-Moraxella clusters were relatively frequently identified at the completion of slaughter, very few cultures were detected on spoiled carcasses . It appears, therefore, that this group of organisms has only low spoilage potential.

J Clin Invest, 1992 Jul, 90(1), 97 - 106
Interactions of surfactant protein D with bacterial lipopolysaccharides . Surfactant protein D is an Escherichia coli-binding protein in bronchoalveolar lavage; Kuan SF et al.; Surfactant protein D (SP-D) is a collagenous glycoprotein that is secreted into the pulmonary airspaces by alveolar type II and nonciliated bronchiolar cells . SP-D exhibits Ca(++)-dependent carbohydrate binding in vitro and is structurally related to the collagenous C-type lectins, including serum conglutinin, serum mannose-binding proteins, and surfactant protein A . Preliminary studies showed calcium- and saccharide-dependent binding of fluorescein-conjugated or radioiodinated SP-D to a variety of microorganisms, including Gram-negative bacteria and fungi . A laboratory strain of Escherichia coli (Y1088) was chosen to further examine the mechanism(s) of binding . Binding of SP-D to Y1088 was time dependent, saturable, and inhibited by cold SP-D or competing saccharides; Scatchard analysis gave a Kd of 2 x 10(-11) M . At higher concentrations, SP-D also caused Ca(++)-dependent agglutination of Y1088 that was inhibited by alpha-glucosyl-containing saccharides, antisera to the carbohydrate-binding domain of SP-D, or Y1088 LPS . Lectin blots showed specific binding of 125I-SP-D to Y1088 LPS, as well as LPS from other several strains of enteric Gram-negative bacteria . Immunogold studies demonstrated strong and uniform surface labeling of the bacteria . Rat and human bronchoalveolar lavage (BAL) caused Ca(++)-dependent agglutination of E . coli that was dose dependent and inhibited by competing saccharides or anti-SP-D . SP-D was selectively and efficiently adsorbed from rat BAL by incubation with E . coli, and incubation of E . coli with radiolabeled rat type II cell medium revealed that SP-D is the major E . coli-binding protein secreted by freshly isolated cells in culture . We suggest that SP-D plays important roles in the lung's defense against Gram-negative bacteria.

Br J Ophthalmol, 1992 Jul, 76(7), 424 - 7
Possible environmental sources of Acanthamoeba spp in contact lens wearers; Seal D et al.; The water supply and dust samples from the home environment (bathrooms and kitchens) of 50 wearers of contact lenses (CLs) were cultured for the presence of free-living amoebae . CL cases, solutions, and water taps were cultured for bacteria, which amoebae require for growth . Acanthamoeba spp were isolated from water drawn from six bathroom cold water taps (tank supplied), five in the presence of limescale, and from one kitchen cold water tap (mains supplied) . There was an association between the presence of limescale in water and direct culture for free-living amoebae, suggesting that scale provides a favourable microenvironment for amoebae . Acanthamoebae were also found in dust from around one washbasin . Nineteen of 50 CL cases, 12/122 CL care rinsing solutions, and 59/100 cold water taps yielded Gram negative bacteria which could be ingested by amoebae . It is concluded from this study that CLs should not be washed in first-drawn tank-fed cold water, especially if limescale is present, and that soft CLs should be rinsed in manufactured single-use, sterile solutions . Rigid CL and CL cases should only be washed with boiled tap water (preferably hot), or single-use sterile solutions, and stored dry to prevent multiplication of amoebae and Gram negative bacteria.

Infect Immun, 1992 Jul, 60(7), 2769 - 76
Legionella pneumophila lipopolysaccharide activates the classical complement pathway; Mintz CS et al.; Legionella pneumophila is a gram-negative bacterium capable of entering and growing in alveolar macrophages and monocytes . Complement and complement receptors are important in the uptake of L . pneumophila by human mononuclear phagocytes . The surface molecules of L . pneumophila that activate the complement system are unknown . To identify these factors, we investigated the effects of L . pneumophila lipopolysaccharide (LPS) on the classical and alternative complement pathways of normal human serum by functional hemolytic assays . Although incubation of LPS in normal human serum at 37 degrees C resulted in the activation of both pathways, complement activation proceeded primarily through the classical pathway . Activation of the classical pathway by LPS was dependent on natural antibodies of the immunoglobulin M class that were present in various quantities in sera from different normal individuals but were absent in an immunoglobulin-deficient serum obtained from an agammaglobulinemic patient . Additional studies using sheep erythrocytes coated with LPS suggested that the antibodies recognized antigenic sites in the carbohydrate portion of LPS . The ability of LPS to interact with the complement system suggests a role for LPS in the uptake of L . pneumophila by mononuclear phagocytes.

Clin Orthop, 1992 Jul, (280), 175 - 8
Case report of an unusual cause of low back pain . Intervertebral diskitis caused by Eikenella corrodens; Noordeen MH et al.; A 42-year-old man suffered from low back pain caused by intervertebral diskitis . The condition was diagnosed on plain roentgenogram and computed tomography scans . The infected area was biopsied and grew Eikenella corrodens, a gram-negative anaerobic rod, which grows slowly only after culture on blood agar in 5-10% carbon dioxide . This is the first reported case of E . corrodens intervertebral diskitis causing acute low back pain . Recognition of this organism in this condition emphasizes the importance of aerobic and anaerobic cultures of infected disks . The organism has unusual antibiotic sensitivities . The infection, once appropriately treated, responded rapidly.

Am J Surg Pathol, 1992 Jul, 16(7), 650 - 7
Immunocytochemical identification of Rochalimaea henselae in bacillary (epithelioid) angiomatosis, parenchymal bacillary peliosis, and persistent fever with bacteremia; Reed JA et al.; We report the immunocytochemical identification of Rochalimaea henselae, a newly recognized fastidious, Gram-negative, Warthin-Starry-positive organism, as the common pathogen in bacillary angiomatosis (BA), bacillary peliosis (BP) of the liver and spleen, and persistent fever with bacteremia in immunocompromised patients . Immunogenic proteins of the R . henselae strain isolated from the blood of a febrile immunocompromised patient with BP of the liver were used to produce primary immune serum in rabbits . Using immunocytochemical procedures, the polyclonal antiserum reacted strongly not only with the immunizing strain of the bacteria, but also with other blood isolates of R . henselae (five cases) from both immunocompromised and immunocompetent patients and with the organisms present in the tissue lesions of cutaneous BA (five cases) and BP of the liver (two cases) and spleen (one case) . The blood isolates and BA and BP tissue samples were obtained from widely separated geographic areas . The antiserum was weakly cross-reactive with cultures of Rochalimaea quintana, an organism closely related to R . henselae, but this reactivity was eliminated by specific adsorption . The antiserum did not cross-react with the Warthin-Starry-positive organisms associated with cat scratch disease (Afipia felis), syphilis (Treponema pallidum), Lyme disease (Borrelia burgdorferi) or chronic active gastritis (Helicobacter pylori) . Likewise, the antiserum did not identify organisms in eight cases of Kaposi's sarcoma, a disorder of immunocompromised patients that is clinically similar to BA . Further studies are needed to determine the prevalence of this newly recognized organism as well as its possible involvement in other angioproliferative diseases.

Mikrobiyol Bul, 1992 Jul, 26(3), 233 - 41
{In vitro activity of various antibiotic agents against gram-negative nosocomial bacteremia pathogens}; Gur D et al.; Empiric therapy is essential in Gram-negative nosocomial bacteremia . Therapy is initiated according to the most frequently isolated organisms and their susceptibility profiles . In order to determine the appropriate antibiotics for this purpose, a study was conducted in 176 blood isolates in Hacettepe University Hospitals . Antibiotic susceptibilities were determined by microbroth dilution tests . The results showed amikacin, ofloxacin and ciprofloxacin to be the most effective antibiotics in-vitro . It is suggested that these agents should be employed in empirical therapy until the causative agent is defined.

Leber Magen Darm, 1992 Jul, 22(4), 156 - 9
{Spontaneous bacterial peritonitis in a patient with decompensated alcoholic cirrhosis of the liver}; Nechwatal R et al.; Spontaneous bacterial peritonitis (SBP) constitute a rare complication of cirrhosis of the liver with a high mortality rate . Unspecific sings of decompensation like increasing elaboration of ascites or the development of hepatic encephalopathy are often prevailing . Fever and peripheral blood leukocytosis as well as abdominal sings of peritonitis are not rarely completely missing . There is a strong suspicion of SBP with an ascitic polymorphonuclear count of greater than 500/mm3 . Determination of lactate and pH in the ascitic fluid can increase diagnostic sensitivity and specificity if the leukocyte count is dubious . Gram negative rods predominate with over 70% whereas anaerobic organisms appear to be very rare . A 49 year old patient is presented and diagnostic and therapeutic strategies reviewed . In view of the high mortality every clinical deterioration of patients with cirrhosis should alert the physician of the presence of SBP.

Vet Pathol, 1992 Jul, 29(4), 269 - 77
Microscopic and ultrastructural studies of necrotizing hepatopancreatitis in Pacific white shrimp (Penaeus vannamei) cultured in Texas; Frelier PF et al.; Granulomatous hepatopancreatitis of unknown etiology has been considered an important disease of Texas shrimp mariculture since 1985 . Samples of Pacific white shrimp (Penaeus vannamei) were collected during 1986, 1987, and 1990 from three farms and an experimental mariculture facility with histories of production loss and increased mortality rates . Histologic and ultrastructural examination of shrimp from the four sites demonstrated two morphologically distinct, Gram-negative, double-enveloped, intracytoplasmic bacteria in necrotic hepatopancreatic epithelium . The more numerous small, pleomorphic rod as well as the helical organism are both taxonomically unclassifiable . The helical organism lacked ultrastructural characteristics of previously described helical or spiral bacteria . The relationship between the two organisms is unknown, but the pleomorphic rod is thought to play a major role in the disease . The role of a bacterial agent(s) in subsequent disease episodes is suggested by the observation that the use of oxytetracycline-medicated feed resulted in increased production and survival . Hepatopancreatic tubular epithelial necrosis and shrimp mortality correlated directly with the extent of infection by the small pleomorphic rod . Individual discrete bacteria were identified microscopically by Steiner and Steiner's method . Three major developmental stages of the disease were characterized based on the extent and number of hepatopancreatic tubular epithelial cells containing bacteria, the degree of tubular interstitial inflammation, and the extent and chronicity of tubular necrosis . Additional studies are needed to clarify the roles of the different bacteria identified and the potential role of environmental factors on the disease process.

Antimicrob Agents Chemother, 1992 Jul, 36(7), 1499 - 503
Comparison of 5 milligrams of netilmicin per kilogram of body weight once daily versus 2 milligrams per kilogram thrice daily for treatment of gram-negative pyelonephritis in children; Vigano A et al.; The efficacy and safety of netilmicin, 5 mg/kg of body weight once daily or 2 mg/kg thrice daily for 10 days, for the treatment of gram-negative pyelonephritis in children were compared in a prospective, randomized trial . Explicit criteria were used to define the site of infection, treatment outcome, and adverse effects . Netilmicin was given to 74 children once daily and to 70 children three times daily . At 1 week posttreatment, 73 (99%) of 74 children treated with netilmicin once daily and 70 (100%) of 70 children treated with netilmicin three times daily were cured . At 4 weeks posttreatment, no relapse was detected and the rate of reinfection was essentially identical in the two treatment groups . Peak serum netilmicin concentrations were higher in patients given the once-daily regimen, whereas a higher trough level was detected in patients given the three-times-daily regimen . Nephrotoxicity, which was defined as an increase in the serum creatinine level of greater than or equal to 0.3 mg/dl over the baseline, was rare (3%) and reversible and occurred regardless of the treatment regimen . Ototoxicity, which was assessed by pure-tone audiometry (250 to 8,000 Hz) and brain stem-evoked response (6,000 Hz), occurred in 2 of 32 children who were evaluated . In these two children, who were given the once-daily regimen, wave V was not evokable monolaterally below 25 and 40 dB normal hearing level, respectively . Thus, it may be possible to treat childhood pyelonephritis with netilmicin once daily . However, this new approach needs to be confirmed in other studies.

Int J Syst Bacteriol, 1992 Jul, 42(3), 451 - 8
Flexibacter ovolyticus sp . nov., a pathogen of eggs and larvae of Atlantic halibut, Hippoglossus hippoglossus L; Hansen GH et al.; A psychrotrophic Flexibacter sp., Flexibacter ovolyticus sp . nov., was isolated from the adherent bacterial epiflora of Atlantic halibut (Hippoglossus hippoglossus L.) eggs and was shown to be an opportunistic pathogen for halibut eggs and larvae . The strains which we isolated had the enzymatic capacity to dissolve both the chorion and the zona radiata of the egg shells . A total of 35 isolates were characterized by using morphological and biochemical tests . These strains were rod shaped, gram negative, Kovacs oxidase positive, and pale yellow and exhibited gliding motility . They did not produce acid from any of the wide range of carbohydrates tested . Our isolates had the ability to degrade gelatin, tyrosine, DNA, and Tween 80 . Starch, cellulose, and chitin were not degraded . The strains were catalase and nitrate reductase positive, did not produce H2S, and did not grow under anaerobic conditions . F . ovolyticus resembles Flexibacter maritimus, but differs from the latter species in several biochemical and physiological characteristics . DNAs from F . ovolyticus strains had guanine-plus-cytosine contents which ranged from 30.3 to 32.0 mol% (strains EKC001, EKD002T {T = type strain}, and VKB004), and DNA-DNA hybridization studies revealed levels of relatedness between F . ovolyticus EKD002T and F . maritimus NCMB 2154T and NCMB 2153 of 42.7 and 30.0%, respectively . Compared with previously described Cytophaga and Flexibacter spp . with low guanine-plus-cytosine contents, F . ovolyticus constitutes a new species . Strain EKD002 (= NCIMB 13127) is the type strain of the new species.

Acta Gastroenterol Belg, 1992 Jul-Aug, 55(4), 333 - 40
{Prevention of bacterial infection using selective intestinal decontamination in patients with cirrhosis admitted to intensive care . Controlled study in 120 patients}; Henrion J et al.; All cirrhotic patients admitted on a medical intensive care unit, were included in a randomized trial of selective intestinal decontamination provided there was no infection on admission . The selective intestinal decontamination consisted of a regimen of 3 oral, nonabsorbable antibiotics for the 74 first patients (Neomycin 1 gr, Colistin 1.500.000 U, Nystatin 1.000.000 U, every 6 hours), then of norfloxacin, 400 mg BID for the following patients . The duration of treatment was at least 5 days . Of the 120 patients, initially randomized to receive or not the treatment, 26 were ultimately excluded, mainly (18 cases) because of infection present but unrecognized at the time of admission . Ninety four patients were thus compared for the efficiency of the treatment, 45 in the treated group and 49 in the not treated group . The results showed a significant reduction of the episodes of septicemia in the treated group (4 versus 12, P = 0.044) . This reduction was evident only for septicemia due to gram negative germs . Mortality was unaffected . When the risk factors were studied, bacterial infection was linked to the degree of hepatic failure . We recommend selective intestinal decontamination for cirrhotic patients admitted on intensive care unit, particularly when hepatic function is poor.

Wiad Lek, 1992 Jul, 45(13-14), 514 - 8
{Lyme disease--also a childhood disease}; Wygledowska G; In the light of a survey of the world literature Lyme borreliosis, an epidemic disease caused by gram-negative organisms Borrelia burgdorferi is described . The disease has three phases with involvement of the skin, the nervous system, the cardiovascular system and joints . Treatment with antibiotics is effective . The disease lies in the fields of interest of various specialties including paediatrics.

Spec Care Dentist, 1992 Jul-Aug, 12(4), 153 - 6
The role of salivary function on oropharyngeal colonization; Gibson G et al.; Aspiration of oropharyngeal flora is the most common route of acquiring Gram-negative pneumonia, a major nosocomial infection . Epidemiologic studies indicate that the elderly are at increased risk for developing these life-threatening pneumonias . The primary objective of this study was to determine the effects of salivary oral defenses against Gram-negative colonization . The study population consisted of 41 male outpatients, age 70 and older, seen at the Denver VAMC . The group included subjects with both diminished and normal salivary flow . Each subject answered a questionnaire regarding overall health, medication use, and symptoms of salivary dysfunction . We then collected whole saliva, unstimulated and stimulated parotid saliva, and performed a throat swab on each patient . For each throat culture, analysis was done to identify Gram-negative bacteria . Flow rates between colonized (n = 6) and noncolonized subjects (n = 34) were compared . While there were no significant differences in the flow rates between the two groups, a trend was noticed in that flow rates were lower for all three flow measures in the colonized group (whole, 22% decrease; unstimulated, 22% decrease; and stimulated, 28% decrease) . These preliminary findings suggest that subjects with diminished salivary flow may possibly be at increased risk for oropharyngeal Gram-negative colonization . Research on the role that saliva plays in oropharyngeal bacterial colonization is continuing.

Indian Pediatr, 1992 Jul, 29(7), 871 - 4
Chest radiographs in neonatal septicemia; Faridi MM et al.; Chest radiographs of 63 culture proven cases of neonatal septicemia were evaluated in this prospective study . Gram negative septicemia was responsible for 76.2% cases . Radiological abnormalities were observed in 27 cases (42.8%) . Seven of these had no respiratory distress . The findings were right sided infiltrates (27%); hyperinflation (7.9%), bronchopneumonia (6.3%) and pneumothorax (1.6%) . Increasing gestational age, late onset of illness (greater than 3 days) and presence of respiratory signs of distress had a positive correlation with presence of X-ray findings . Term newborns with respiratory distress of late onset sepsis (greater than 3 days) had significantly higher number (p less than 0.05) of abnormal radiographs . Presence of radiological abnormality neither influenced the clinical outcome nor was affected by the causative organisms . The practice of doing a chest radiograph routinely in cases of neonatal septicemia is justified irrespective of presence of respiratory signs of distress.

Can J Microbiol, 1992 Jul, 38(7), 614 - 7
Binding of mutagenic pyrolyzates to fractions of intestinal bacterial cells; Zhang XB et al.; The binding of mutagenic pyrolyzates to cell fractions from some gram-negative intestinal bacteria and to thermally treated bacterial cells was investigated . 3-Amino-1,4-dimethyl-5H-pyrido{4,3-b}indole (Trp-P-1) and 3-amino-1-methyl-5H-pyrido{4,3-b}indole (Trp-P-2) were effectively bound by several of the bacterial cells . The cell wall skeletons of all bacteria effectively bound Trp-P-1 and Trp-P-2 . Their cytoplasmic fractions retained Trp-P-1 and Trp-P-2, but to a lesser extent than the cell wall skeletons . 2-Amino-3-methylimidazo{4,5-f}quinoline (IQ) was not found in their cytoplasmic fractions . These cell wall skeletons also bound 2-amino-6-methyldipyrido{1,2-a:3',2'-d}imidazole (Glu-P-1), 2-amino-5-phenylpyridine (Phe-P-1), IQ, 2-amino-3,4-dimethylimidazo{4,5-f}quinoline (MeIQ), and 2-amino-3,8-dimethylimidazo{4,5-f}quinoxaline (MeIQX) . The amount of each mutagen bound differed with the type of mutagen and the bacterial strain used . The outer membrane of Escherichia coli IFO 14249 showed binding of about 123.7 micrograms/mg of Trp-P-2, and its cytoplasmic membrane bound 57.14 micrograms/mg . Trp-P-2 bound to the bacterial cells was extracted with ammonia (5%), methanol, and ethanol but not with water.

Immunol Today, 1992 Jul, 13(7), 271 - 6
Lipopolysaccharide antagonists; Lynn WA et al.; Bacterial lipopolysaccharide (LPS) is a potent and pleiotropic stimulus of immune cells . LPS has important clinical relevance because it has a direct role in the pathogenesis of Gram-negative bacterial infection . The lipid A moiety of LPS is responsible for the toxic effects of LPS . The identification of structural analogs and precursors of lipid A, which are apparently competitive antagonists of the biological actions of LPS, is strong evidence that the actions of LPS are mediated by a specific LPS receptor or family of receptors . Identification and analysis of these LPS receptors with LPS antagonists should help to define the pathways of cellular activation by LPS and lead to the development of novel anti-LPS strategies in the therapy of bacterial diseases.

Proc Natl Acad Sci U S A, 1992 Jul 1, 89(13), 5735 - 9
In vivo production of a stable single-stranded cDNA in Saccharomyces cerevisiae by means of a bacterial retron; Miyata S et al.; Gram-negative bacteria such as Myxococcus xanthus, Stigmatella aurantiaca, and Escherichia coli contain retroelements called retrons . Retrons consist of the msr-msd region and the gene for reverse transcriptase (RT), which are essential for the production of the branched RNA-linked ms-DNA (multicopy single-stranded DNA) . In this study, we attempted to produce msDNA in the yeast Saccharomyces cerevisiae . Retron Ec67 from E . coli, which is responsible for the production of msDNA-Ec67, was cloned under the GAL10 promoter in a 2-microns-based plasmid . msDNA thus produced was detected by extending the 3' end of the msDNA by avian myeloblastosis virus RT . This yielded a main product of 117 nucleotides . Treatment of this product with RNase A resulted in a DNA of 105 nucleotides . These results are in good agreement with the structure of msDNA-Ec67 . The production of msDNA-Ec67 was further confirmed by Southern blot hybridization . The msDNA production was dependent upon the bacterial RT gene in the clone and was increased severalfold when the RT gene of retron Ec67 was placed in front of the msr-msd region . The potential of msDNA as a eukaryotic vector producing a stable single-stranded DNA as well as RNA is discussed.

Am J Respir Cell Mol Biol, 1992 Jul, 7(1), 81 - 9
Endotoxin enhancement of lymphocyte adherence to cultured sheep lung microvascular endothelial cells; Jones M et al.; The most common predisposing factor for development of the adult respiratory distress syndrome is gram-negative sepsis . Our previous studies have shown that a single infusion of Escherichia coli endotoxin into sheep causes early sequestration of lymphocytes in the lungs' microcirculation . In this report, we examined the effects of endotoxin on sheep lymphocyte adherence to sheep pulmonary microvascular endothelial cells in vitro . Endothelial cells were exposed to endotoxin, and subsequent adherence of 51Cr-labeled lymphocytes was measured in a monolayer adhesion assay . Endotoxin enhanced adherence of lymphocytes isolated from blood and caudal mediastinal node (CMN) lymph in a time- and dose-dependent manner . Adherence of CMN lymphocytes increased from a control value of 13.6 +/- 1.6% to 29.9 +/- 3.1% after 4 h of treatment with 1 microgram/ml endotoxin . Both B and T lymphocytes contributed to the increased adherence . Pretreatment of the endothelial cells with cycloheximide revealed that the endotoxin-enhanced adherence was partially dependent upon protein synthesis . Morphologic studies revealed that enhanced adherence was accompanied by a 5-fold increase in migration of lymphocytes between endothelial cells . In contrast to human umbilical vein endothelial cells, antibodies to the known lymphocyte adherence molecules, lymphocyte function-associated antigen (LFA-1), CD-44, and the lymphocyte homing receptor (LECAM-1), were ineffective in blocking adherence to the sheep pulmonary endothelial cells . We conclude that the acute sequestration of lymphocytes in the pulmonary microcirculation of sheep after endotoxin administration is due to increased adhesive properties of the endothelial cells . Our data suggest that this adherence is mediated by as yet undescribed mechanisms that may be unique to pulmonary microvascular endothelium.

Immunol Lett, 1992 Jul, 33(2), 135 - 8
Murine ascitic fluids contain varying amounts of an inhibitor that interferes with complement-mediated effector functions of monoclonal antibodies; Appelmelk BJ et al.; The ability of murine monoclonal antibodies (mAbs), directed to the inner core of Gram-negative bacterial lipopolysaccharide (LPS, endotoxin), to enhance complement-mediated killing of bacteria, was investigated . The mAbs were tested as present in ascitic fluid . It was found that ascites contains an factor that inhibited the activity of complement . This effect was evident in assays for complement-mediated lysis of antibody-coated Gram-negative bacteria (bacterial killing) or of opsonised red blood cells . Moreover, the amount of inhibitor was found to vary from one ascites to another and spanned a 60-fold range . Thus, in vitro or in vivo experiments where complement is known to play a determining role may yield incorrect results when ascites is used as a source of antibody; the use of ascites prepared from irrelevant antibody as a negative control does not eliminate this problem.

Scand J Gastroenterol, 1992 Jun, 27(6), 453 - 8
Quantification of lipopolysaccharides in human bile with or without gram-negative bacteria; Osnes T et al.; Lipopolysaccharide (LPS) was assayed in 78 samples of human common-duct bile, obtained at endoscopic retrograde cholangiography . The LPS was assayed by a chromogenic limulus amebocyte lysate (LAL) test, after dilution of bile samples in heparinized plasma and inactivation of inhibitors . The assay was not influenced by other biliary constituents, as demonstrated by the recovery of standards . Bile pigments did not influence the results . The LAL test was positive in 60 of the samples, 59 of which had a positive culture for gram-negative bacteria or Candida sp . The levels of LPS were significantly correlated to the total number of bacteria (n = 16, R = 0.55, p less than 0.05) . The median LPS level was 35,250 ng/l and showed a very large variation (140 ng/l to 27.8 mg/l) . In four of the samples gram-negative bacteria were present, but no LPS could be detected . The study demonstrates the presence of LPS in great quantities in human bile and supports the feasibility of using the LAL test on bile samples . The presence of LPS (within the detection limit) appears to be associated with local microbial colonization.

Eur Respir J, 1992 Jun, 5(6), 675 - 9
Branhamella catarrhalis respiratory infections; Barreiro B et al.; Branhamella catarrhalis is an aerobic Gram-negative diplococcus . It has been traditionally regarded as an oropharyngeal commensal and until recently was only identified as a pathogen in cases of bronchopulmonary infections . The aim of this study was to analyse the characteristics of the respiratory infections caused by B . catarrhalis and to know the antibiotic susceptibility of this microorganism . We retrospectively studied 32 lower respiratory tract infections, caused by B . catarrhalis (20 cases of bronchial infection and 12 cases of pneumonia), diagnosed between 1988-1989 in our hospital . All patients had an underlying disease; chronic obstructive pulmonary disease (COPD) and chronic heart disease being the most frequent . The aetiological diagnostic procedures were: sputum culture in 28 cases (15 in pure culture and 13 mixed), protected specimen brush (PSB) in three cases and transthoracic needle aspiration (TNA) in one case . Twenty B . catarrhalis isolates were penicillin and ampicillin-resistant, 11 in the pneumonia group and 9 in the bronchial infection group . All isolates were sensitive to amoxycillin-clavulanic acid and second generation cephalosporin . In our group four patients died . We conclude that B . catarrhalis is a not infrequent cause of respiratory infection, particularly in COPD patients, and that the high incidence of antibiotic resistance to penicillin and ampicillin should be taken into account before considering an empirical antibiotic treatment.

Clin Infect Dis, 1992 Jun, 14(6), 1213 - 28
Gram-negative sepsis and the adult respiratory distress syndrome; Martin MA et al.; Gram-negative sepsis has dramatically increased in frequency throughout the twentieth century in the United States . Currently, approximately 200,000 patients develop gram-negative sepsis each year in this country . Of these, about one-quarter develop the adult respiratory distress syndrome (ARDS) . Among these critically ill patients, mortality is estimated at 60%-90% . In the complex series of events leading to acute lung injury in gram-negative sepsis, endotoxin is the proximal mediator . Although endotoxin may be capable of causing direct injury to the pulmonary endothelium, its primary role is as a trigger activating inflammatory agents, including complement, neutrophils, and platelets, and inducing the production of cytokines and arachidonic acid metabolites . The end results are impairment of the endothelial barrier, diffusely increased capillary permeability, and adherence of neutrophils to the endothelium with subsequent migration into the tissues . The consequent clinical syndrome is one of acute respiratory distress with pulmonary edema, poorly compliant lungs, and refractory hypoxemia . Endothelial injury often becomes widespread, leading to the failure of multiple organs, including the kidneys, brain, intestine, and liver . Conventional therapy consists of supplemental oxygen, positive end-expiratory pressure, inotropic agents, fluid management, and antibiotics aimed at the offending pathogen . Recent discoveries regarding the mediators of sepsis as well as the expansion of the biotechnological armamentarium have provided clinicians with a plethora of new tools with which to manipulate the host's inflammatory response . The challenge for the next decade will be to ensure the safety, efficacy, and cost-effective use of these expensive but potentially lifesaving immunomodulators, singly or in combination, as adjuvant therapy.

Crit Care Med, 1992 Jun, 20(6), 746 - 50
Septic shock in patients with cirrhosis: hemodynamic and metabolic characteristics and intensive care unit outcome; Moreau R et al.; OBJECTIVES: To examine the hemodynamic and metabolic characteristics and ICU outcome of septic shock in patients with cirrhosis . DESIGN: Prospective, comparative study . Measurements performed in the first 24 hrs of septic shock . SETTING: A general hospital ICU . PATIENTS: Twelve patients with cirrhosis and 23 patients without cirrhosis admitted for septic shock . MEASUREMENTS AND MAIN RESULTS: Arterial pressure was measured using an arterial catheter . Pulmonary arterial and right atrial pressures were measured by using a pulmonary artery catheter . Cardiac output was determined by using the thermodilution method . Pulmonary arterial L-lactate plasma concentrations were measured using an automated spectrophotometer, and blood temperature was measured using a cardiac output computer . Arterial and mixed venous PO2, PCO2, and pH values were measured by using specific electrodes . Oxygen saturations and hemoglobin concentrations were measured using a hemoximeter . Patients with cirrhosis had decompensated liver disease (grade C of the Child-Pugh classification) . The number of Gram-negative infections and therapeutic interventions were similar in both groups . Patients with cirrhosis had higher cardiac indices (5.14 +/- 0.52 {SE} vs . 3.91 +/- 0.30 L/min/m2, p less than .05), plasma lactate concentrations (9.0 +/- 2.0 vs . 5.2 +/- 0.7 mmol/L, p less than .05) and ICU mortality rates (100% vs . 43%, p less than .05), and lower blood temperatures (35.5 +/- 0.6 vs . 37.6 +/- 0.2 degrees C, p less than .05) than patients without cirrhosis . Systemic vascular resistance, arterial pressure, pulmonary arterial pressure, oxygen delivery and consumption, and arterial and mixed venous acid-base status were not significantly different between the two groups . CONCLUSIONS: In patients with cirrhosis, septic shock was characterized by severe liver dysfunction, low blood temperature, marked increases in cardiac index and lactic acidemia, and a 100% ICU mortality rate . These findings should be taken into account if patients with cirrhosis are to be included in controlled studies on septic shock.

J Bacteriol, 1992 Jun, 174(11), 3429 - 38
Localization of cytochromes to the outer membrane of anaerobically grown Shewanella putrefaciens MR-1; Myers CR et al.; In gram-negative bacteria, numerous cell functions, including respiration-linked electron transport, have been ascribed to the cytoplasmic membrane . Gram-negative bacteria which use solid substrates (e.g., oxidized manganese or iron) as terminal electron acceptors for anaerobic respiration are presented with a unique problem: they must somehow establish an electron transport link across the outer membrane between large particulate metal oxides and the electron transport chain in the cytoplasmic membrane . When the metal-reducing bacterium Shewanella putrefaciens MR-1 is grown under anaerobic conditions and membrane fractions are purified from cells lysed by an EDTA-lysozyme-polyoxyethylene cetyl ether (Brij 58) protocol, approximately 80% of its membrane-bound cytochromes are localized in its outer membrane . These outer membrane cytochromes could not be dislodged by treatment with chaotropic agents or by increased concentrations of the nonionic detergent Brij 58, suggesting that they are integral membrane proteins . Cytochrome distribution in cells lysed by a French press protocol confirm the localization of cytochromes to the outer membrane of anaerobically grown cells . This novel cytochrome distribution could play a key role in the anaerobic respiratory capabilities of this bacterium, especially in its ability to mediate manganese and iron reduction.

Infect Immun, 1992 Jun, 60(6), 2506 - 13
Binding and neutralization of endotoxin by Limulus antilipopolysaccharide factor; Warren HS et al.; In order to examine the ability of Limulus antilipopolysaccharide factor (LALF) to bind lipopolysaccharide (LPS), we purified LALF to homogeneity from Limulus amoebocyte lysate and coupled it covalently to agarose beads . LALF-coupled beads captured more tritiated LPS from rough and smooth strains of gram-negative bacteria than did control human serum albumin-coupled beads . Unlabeled homologous and heterologous LPS competed for the binding of 3H-LPS to LALF-coupled beads . LALF bound LPS in a dose-dependent manner as assessed by the precipitation of LPS-LALF complexes with 50% saturated ammonium sulfate . We also studied the ability of LALF to neutralize LPS . LPS preincubated with LALF was less mitogenic for murine splenocytes, was less pyrogenic in the rabbit fever assay, was less lethal in mice which had been sensitized to LPS with actinomycin D, and induced less fever, neutropenia, and pulmonary hypertension when infused into sheep . Our findings extend prior studies which suggested that LALF binds to and neutralizes LPS from multiple strains of gram-negative bacteria.

Infect Immun, 1992 Jun, 60(6), 2281 - 7
Intracellular growth of Afipia felis, a putative etiologic agent of cat scratch disease; Birkness KA et al.; The organism Afipia felis, which is though to be an etiologic agent of cat scratch disease, is a gram-negative rod that is clearly seen in infected tissue but is very difficult to isolate from clinical specimens; there has been only one report to date of the successful isolation and maintenance of the bacterium on artificial medium . We have found that A . felis will attach, invade via phagocytosis, and multiply intracellularly within the phagosomes of primary human monocytes and HeLa cells . Once in the cell, the bacterium appears to change morphologically, becoming longer and more pleomorphic, and loses its ability to grow on an artificial medium . Unique proteins have been identified in both the intra- and extracellular variants of A . felis . Convalescent-phase sera from patients with cat scratch disease react poorly with intracellular and extracellular bacteria, suggesting a poor humoral response . The tissue culture protocol presented has been used to isolate 14 new strains of A . felis and has for the first time permitted study of the pathogenesis of this unique organism.

J Infect Dis, 1992 Jun, 165(6), 1138 - 41
Interactions between live Bartonella bacilliformis and endothelial cells; Garcia FU et al.; Bartonella bacilliformis, a gram-negative, flagellated, motile bacterium, is the etiologic agent of verruca peruana . It is found within the verruca, where it can form large cytoplasmic (Rocha-Lima) inclusions in endothelial cells . Previously, an activity has been described in homogenates of B . bacilliformis that in vitro increases the proliferation of endothelial cells and their production of tissue-type plasminogen activator (t-PA) and in vivo is angiogenic . The aim of the present study was to determine if live B . bacilliformis similarly stimulated endothelial cells and produced the Rocha-Lima inclusion . By measuring proliferation of cells and the production of t-PA in vitro, it was found that the live bacteria increased both parameters in a fashion similar to the homogenates of B . bacilliformis . Interaction between the bacteria and endothelial cells appeared to be necessary for proliferation . On electron microscopy, bacteria penetrated the endothelial cell within 1 h, forming a small membrane-bound inclusion . By 12 h, a large membrane-bound inclusion, similar to the Rocha-Lima inclusion, containing numerous bacteria was present . These data provide further evidence that B . bacilliformis has an angiogenic activity and that the bacteria are at least in part responsible for the vascular proliferation of the verruca.

Circ Shock, 1992 Jun, 37(2), 164 - 8
Differential contractile responses of mesenteric and pulmonary artery segments to norepinephrine and phorbol ester in the septic pig; Suba EA et al.; The contractile response of isolated vascular segments was studied in Yucatan miniature swine approximately 48 hr after induction of sepsis by intraperitoneal injection of live Escherichia coli . Compared to non-septic controls, segments of the cranial mesenteric artery from septic animals showed a significantly attenuated contractile response to the adrenergic receptor agonist norepinephrine (NE) . The EC50 for NE increased from 1.1 +/- 0.3 to 6.3 +/- 2.0 microM and the Emax decreased from 1,010 +/- 179 to 387 +/- 75 mg tension/mg tissue . In contrast, segments of the pulmonary artery showed no significant difference in contractility to NE between sham-operated and septic animals . Mesenteric and pulmonary artery segments from both septic and control animals exhibited similar contraction to the protein kinase C activator phorbol-12, 13-dibutyrate . This suggests that the observed hyporeactivity to NE in porcine mesenteric artery segments is not simply due to cellular damage by toxins associated with the septic state . The results also indicate that the impact of gram-negative sepsis on vascular contractile function varies between tissue from the systemic and pulmonary circulation in pigs.

Circ Shock, 1992 Jun, 37(2), 111 - 6
Protein kinase C activity and lipogenesis from glucose in isolated adipocytes of endotoxemic rats; Spitzer JA et al.; The hypothesis was tested that the insulin-like effects of Gram-negative bacterial endotoxins (ET), exerted after in vivo administration on subsequently isolated adipocytes, might be associated with changes in protein kinase C (PKC) activity . The latter is believed to be involved in insulin's mechanism of action on adipocytes . E . coli ET was administered to rats either as a bolus injection (1 mg/100 g bw, in 160-180 g rats, LD50 for 6 hr) or via subcutaneously implanted osmotic pumps (0.1 mg/100 g bw/24 hr, for 30 hr, in 340-380 g rats) . Control animals received sterile saline . At 6 hr after bolus injection, and at 30 hr after the onset of ET infusion, the animals were sacrificed and epididymal adipocytes isolated . PKC activity and intracellular distribution were assayed after partial purification on DE-52 cellulose minicolumns . Lipogenesis was measured by {3-3H}-D-glucose incorporation into triglyceride . ET treatment by either mode induced a significant increase (76-80%) in PKC activity . PKC intracellular distribution was altered only in chronically ET-treated rats and was expressed as an increased enzyme activity in the membrane fraction . The increased PKC activity was associated with elevated rates of insulin-stimulated lipogenesis only in young rats . We conclude that in young rats, whose adipocytes display high rates of lipogenesis along with elevated insulin sensitivity, PKC is likely to be one of the possible factors involved in mediation of insulin-like effects of ET.

Eur J Immunol, 1992 Jun, 22(6), 1663 - 5
Lipopolysaccharide induces up-regulation of CD14 molecule on monocytes in human whole blood; Marchant A et al.; We examined by flow cytometry the expression of lipopolysaccharide (LPS) receptor CD14 molecule on monocytes after addition of LPS to human whole blood . Within 30 min LPS induced an increase in monocyte CD14 expression, peaking between 1 and 3 h and followed by a slow decrease . Maximal increase in anti-CD14 monoclonal antibody binding sites was estimated as twofold the basal value . This effect, already observed with very low concentrations of LPS (10 pg/ml), was dose dependent . Protein synthesis was not involved in the CD14 hyperexpression since it was not influenced by co-incubation with cycloheximide . Finally, LPS-induced up-regulation of monocyte CD14 was associated with an increased binding of fluoresceinated LPS . We conclude that LPS in whole blood up-regulates the expression of its own CD14 receptor on monocytes, a phenomenon that could be relevant to the pathogenesis of gram-negative sepsis.

Res Microbiol, 1992 Jun, 143(5), 471 - 9
Transposon mutagenesis in Legionella pneumophila . I.--Persistence of suicide and broad host-range plasmids; Tully M et al.; Two of three highly virulent strains of Legionella pneumophila could act as recipients at high frequencies in conjugation experiments with Escherichia coli donor strains carrying broad host-range plasmids belonging to incompatibility groups N, P and W . All broad host-range and most transposon-delivery plasmids persisted within transconjugants with high stability . Only one (pSUP1021) of several vehicles designed for the delivery of transposons into the chromosome of Gram-negative bacteria was found to yield transposon mutants of Legionella at a detectable frequency.

Ann Hematol, 1992 Jun, 64 Suppl, A158 - 61
Prevention and treatment of CMV infection after allogeneic bone marrow transplant; Gluckman E et al.; CMV infection is the major infectious complication following bone marrow transplantation . It is most often related to reactivation of latent infection in patients who were CMV seropositive before BMT . The incidence and severity have recently been modified by the use of preventive and curative treatments . Prevention of CMV infection with the transfusion of seronegative blood products is useful only when donor and recipient are seronegative . High-dose acyclovir has been shown effective in one randomized study . A multicenter study is currently being performed in Europe to confirm this result . Intravenous gammaglobulins seemed to lower the number of patients who incur interstitial pneumonitis but not the incidence of viremia . They also decreased the incidence of gram-negative sepsis and severe GVH and improved survival . The treatment is based on the use of gancyclovir . Several studies show that gancyclovir is more effective in asymptomatic patients with viral isolation from blood or bronchoalveolar lavage . The addition to gancyclovir of high-dose gammaglobulin improves survival in symptomatic patients with interstitial pneumonitis . This progress in the prevention and treatment of CMV infection has improved the overall results of allogeneic bone marrow transplantation.

Cytokine, 1992 May, 4(3), 205 - 13
Oxidative tryptophan metabolism in renal allograft recipients: increased kynurenine synthesis is associated with inflammation and OKT3 therapy; Holmes EW et al.; Serum concentrations of tryptophan (TRP) and kynurenine (KYN) were determined in renal allograft recipients (RAR) as an index of interferon-gamma-induced, indoleamine-dioxygenase-catalysed TRP degradation . Serum TRP and KYN in RAR during periods of stable graft function were typically within the normal range, however, the median values for serum KYN demonstrated significant increases 5-7 days prior to biopsy-confirmed acute rejection (1.6-fold, P less than 0.01) and on the day of biopsy (1.7-fold, P less than 0.001) . Serum KYN was also markedly elevated in patients who contracted viral or Gram-negative bacterial infections in the absence of graft rejection . Serum KYN was not correlated with serum creatinine in RAR nor were serum TRP or KYN affected by antirejection therapy with high dose steroids . Retrospective analysis of intra-patient changes in serum KYN demonstrated that KYN monitoring was a useful adjunct to serum creatinine in the early detection of first acute rejection episodes . The first course of OKT3 therapy was associated with low serum TRP and significant increases in serum KYN (two- to three-fold) following the first three doses . The time course of these abnormalities corresponded to that over which many of the side effects of the OKT3 'first dose reaction' have been reported to occur . Significant changes in serum KYN were not observed in patients receiving repeat courses of OKT3 therapy . Significant decreases in serum TRP and significant increases in serum KYN were both prevalent and frequent in RAR during the first two postoperative months.(ABSTRACT TRUNCATED AT 250 WORDS)

Br J Anaesth, 1992 May, 68(5), 499 - 502
Gastroduodenal dysfunction as a cause of gastric bacterial overgrowth in patients undergoing mechanical ventilation of the lungs; Inglis TJ et al.; We have studied 15 patients undergoing mechanical ventilation of the lungs for evidence of a relationship between gastroduodenal dysfunction and gastric bacterial overgrowth . Duodenal reflux was detected by quantitative measurement of conjugated bilirubin in gastric aspirate specimens . The pH and bacterial content of these specimens were analysed . A minority of specimens with pH less than 3.5 contained measurable numbers of viable bacteria . The total bacterial count and the count of Gram negative bacteria correlated significantly with specimen pH in both cases (P less than 0.001, in both) . In the 72 specimens of gastric aspirate with pH greater than 3.5, the presence of Gram negative bacteria was associated significantly with detectable bilirubin (P less than 0.001) . The total bacterial count was greater also in specimens containing bilirubin (P = 0.009) . These results suggest that gastroduodenal dysfunction may promote gastric bacterial overgrowth, contributing to the development of ventilator-associated pneumonia.

Klin Monatsbl Augenheilkd, 1992 May, 200(5), 519 - 21
{Acute anterior uveitis: para-infectious hypothesis in predisposed individuals}; Andenmatten R et al.; The etiology and the physiopathology of acute anterior uveitis (AAU) is not well understood yet . However, two major predisposing factors have been identified: a bacterial infection especially with gram negative organisms (functioning as a trigger) and a genetic background, in particular the expression of HLA B-27 tissue antigen . We report the case of a young woman returning from travel to the Far East with her partner . Both presented simultaneously a gastrointestinal infection with fever and diarrhea . Despite extensive investigations, the infectious agent was never identified because of early empirical antibiotic therapy . A few days later, the patient developed AAU of a moderate grade in both eyes . HLA B-27 testing was positive for her, but not for her partner . Experimental research, based on a animal model such as endotoxin-induced uveitis (EIU), gives us some insight into the possible pathogenic mechanisms of AAU . Footpad injection of endotoxin (lipopolysaccharide component of the wall of gram negative), produce an acute anterior uveitis in rats . Extensive histologic analysis of other organs shows that the anterior segment of the eye is the only structure involved . Intensity of inflammation varies in different rat strains, stressing the importance of the genetic background . The similarity of the animal model to AAU will contribute to orient clinical research towards identifying more thoroughly the possible infectious agent at the origin of AAU and possibly to develop a prophylactic therapy.

Childs Nerv Syst, 1992 May, 8(3), 139 - 41
The probable role of hydrocephalus in the development of intraventricular septa . An observation of one case; Lazareff JA et al.; Multiloculated cerebral ventricles are a well-known complication of neonatal meningitis secondary to gram-negative bacteria . Their presence often hampers the proper functioning of the multiple shunts required for draining the ventricles . To determine whether the associated hydrocephalus may contribute to the development of the ventricular septa, we analyzed the histological structure of the ventricular wall and of the ventricular septa of a child who died as a consequence of Escherichia coli meningitis and unilateral hydrocephalus . Our results show that the ventricular septa are formed by glial protrusion into the ventricles . The ependymal wall of the grossly dilated ventricle was disrupted, but not that of the contralateral ventricle . We hypothesize that the presence of an insufficiently controlled hydrocephalus may have been one of the factors that induced the formation of the septa . It is suggested that an external ventricular derivation should be used during the active phase of the infection.

Clin Infect Dis, 1992 May, 14(5), 1089 - 99
beta-Lactamases of gram-negative bacteria: new challenges for new drugs; Sanders CC; The major emphasis in new drug design within the beta-lactam family has been on compounds less susceptible to hydrolysis by beta-lactamases and on combinations containing an enzyme-labile drug plus a beta-lactamase inhibitor . The introduction of such new compounds into clinical use has been followed by the discovery of novel mechanisms of resistance among gram-negative bacteria . These include the appearance of new enzymes, many of which are derivatives of older beta-lactamases . In addition, genes for certain broad-spectrum enzymes previously restricted to chromosomal sites have moved onto plasmids . There is now a greater appreciation of how alterations in enzyme expression--either alone or in concert with changes in drug permeation--can also lead to resistance . Clearly, recent events in the development of new beta-lactam agents have led to a new phase in the understanding of beta-lactam resistance.

Optom Vis Sci, 1992 May, 69(5), 354 - 7
Conjunctival flora in extended wear of rigid gas permeable contact lenses; Fleiszig SM et al.; A longitudinal study was performed to examine the effect of rigid gas permeable (RGP) contact lenses (Boston Equalens II and Quantum II) on the conjunctival flora of 45 young healthy subjects . Microbial flora were determined before delivery of lenses . Subjects wore lenses on an extended wear basis, removing them every 7 days for cleaning and disinfection . Cultures were repeated after 2 months of lens wear and the microbial flora were found to be significantly altered compared to the prelens wear results (0.05 greater than p greater than 0.02) . Changes to conjunctival flora included an increase in the number of eyes from which potentially pathogenic microorganisms were isolated, an increase in the number of eyes that were culture-negative, and a decrease in the number of eyes harboring only normal conjunctival flora . The increase in potentially pathogenic flora was not specific for Gram-negative bacteria, which are most often associated with infectious keratitis during contact lens wear.

J Trauma, 1992 May, 32(5), 564 - 9
Intra-abdominal sepsis and adrenergic receptor response; Forse RA et al.; This study measured the adrenergic receptor response of 13 patients with severe intra-abdominal sepsis, who required laparotomy and an open abdominal closure with Marlex mesh . The source of the sepsis was gram-negative organisms of intestinal origin . There were seven survivors and six nonsurvivors . When the patients were stratified into survivors and nonsurvivors, the Septic Severity Score, the APACHE II score, the Acute Physiological Score, and the Glasgow Coma Scale score results were not significantly different between groups . The alpha-2 and beta-1 adrenergic receptor responses were measured in the adipose tissue of the abdominal wall and the small bowel mesentery on day 1 of admission to the intensive care unit . The results demonstrated that the alpha-2 and beta-1 receptors of the nonsurvivors had a significantly decreased receptor response with desensitization and down regulation . The alpha-2 and beta-1 receptors of the survivors had an increased response with hypersensitization and up regulation . This study indicates that the adrenergic receptor pattern is distinctly different between survivors and nonsurvivors with severe abdominal gram-negative sepsis . The pattern differences occurred early (within 24 hours) when the patients had similar physiologic profiles . It is concluded that adrenergic receptor response may be a biologic indicator of the magnitude of the septic injury and a predictor of outcome.

Clin Pediatr (Phila), 1992 May, 31(5), 295 - 8
A comparison of silver nitrate with erythromycin for prophylaxis against ophthalmia neonatorum; Zanoni D et al.; Although prophylactic eyedrops to treat ophthalmia neonatorum is mandated nationwide, states and hospitals are free to choose specific drugs . To compare two of these agents, we studied the incidence and characteristics of ophthalmia neonatorum in two UCLA teaching hospitals over a five-year period . One, which used 1% silver nitrate solution exclusively, had 50 cases in 34,772 births, a frequency of 0.14% . The other used 0.5% erythromycin ointment exclusively and had 43 cases in 12,652 births, a frequency of 0.34% . Ophthalmia neonatorum was more frequent in the hospital using erythromycin (p less than 0.001), as was chlamydial conjunctivitis (p less than 0.02) . Although not statistically significant, gonococcal conjunctivitis was found in four infants, all in the hospital using silver nitrate . Because silver nitrate was found more effective in decreasing the total frequency of all cases of ophthalmia neonatorum, and cases caused by Chlamydia and gram-negative bacteria specifically, this drug still should be considered as a primary prophylactic agent against ophthalmia neonatorum.

Infect Immun, 1992 May, 60(5), 1946 - 51
Identification and purification of a cpn60 heat shock protein homolog from Helicobacter pylori; Dunn BE et al.; Helicobacter pylori is associated with gastritis and peptic ulcer disease in humans . We have identified a homolog of the chaperonin cpn60 family of heat shock proteins in H . pylori, referred to as Hp54K . Hp54K, purified from water-extractable H . pylori proteins, migrated as a single band at 54 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Its native molecular mass was 740 kDa; thus, Hp54K apparently comprises a 14-mer . The N-terminal 33 residues of Hp54K exhibited 60.6, 57.6, 54.5, 54.5, 51.5, and 51.5% identity with corresponding sequences in the following cpn60 homologs: HtpB (Legionella pneumophila), P1 (human mitochondria), GroEL (Escherichia coli), BA60K (Brucella abortus), HypB (Chlamydia trachomatis), and the 65-kDa immunodominant protein of Mycobacterium bovis BCG, respectively . Hp54K was the only protein recognized in whole-cell preparations of H . pylori by immunoblotting using monospecific antisera against cpn60 homologs from L . pneumophila, E . coli, C . trachomatis, and M . bovis BCG . Antiserum against Hp54K recognized proteins with molecular masses of 50 to 60 kDa in a large number of gram-negative bacteria, consistent with the known highly conserved nature of cpn60 proteins . Hp54K is a major protein and is immunogenic in humans infected with H . pylori . Thus, Hp54K shares many similarities with known cpn60 homologs . On the basis of the proposed role of other cpn60 proteins in induction of chronic inflammation, immune cross-reactivity between Hp54K and gastric tissue may provide an important link between H . pylori infection and gastritis.

Infect Immun, 1992 May, 60(5), 1756 - 60
Effects of endotoxin-associated protein on hematopoiesis; Porat R et al.; Endotoxin-associated protein (EAP), a gram-negative bacterial cell wall component, was evaluated for its effects on hematopoietic colony formation in vitro . Colony-stimulating activity, induced by EAP on circulating and bone marrow progenitor cells, was found to be partially mediated by T cells and augmented by interleukin-3 . The addition of anti-human interleukin-1 (IL-1) antibodies reduced EAP activity, suggesting that EAP may induce IL-1 production . However, EAP was shown to promote the growth of mature progenitor cells independently, unlike the effects of IL-1 on the hematopoietic system . These studies demonstrate that bacterial components other than lipopolysaccharide, such as EAP, may have hematopoietic activity.

Proc Natl Acad Sci U S A, 1992 May 1, 89(9), 4076 - 80
Coordinated antiinflammatory effects of interleukin 4: interleukin 4 suppresses interleukin 1 production but up-regulates gene expression and synthesis of interleukin 1 receptor antagonist; Vannier E et al.; Interleukin 1 receptor antagonist (IL-1ra), a naturally occurring polypeptide with amino acid sequence homology to interleukin 1 alpha (IL-1 alpha) and interleukin 1 beta (IL-1 beta), prevents Escherichia coli-induced shock and death . Both IL-1 and IL-1ra are produced by monocytes stimulated with lipopolysaccharide (LPS) . Because interleukin 4 (IL-4) suppresses IL-1 production, we investigated whether IL-4 modulated IL-1ra synthesis in LPS-stimulated human peripheral blood mononuclear cells . IL-1 beta and IL-1ra were measured by specific RIAs . IL-4 alone (0.01-100 ng/ml) did not stimulate IL-1 beta synthesis but rather induced IL-1ra (4.82 +/- 0.94 ng/ml) . LPS induced synthesis of both IL-1 beta (6.67 +/- 1.06 ng/ml) and IL-1ra (10.77 +/- 2.79 ng/ml) . IL-4 suppressed LPS-induced IL-1 beta mRNA accumulation and synthesis . However, IL-4 acted synergistically with LPS in inducing IL-1ra . IL-4 enhanced LPS-induced IL-1ra mRNA accumulation 4-fold and IL-1ra protein synthesis nearly 2-fold . Moreover, IL-1ra mRNA levels were maximal after 6 hr of exposure to LPS but peaked within the first 3 hr in the presence of IL-4 . IL-4 added as late as 12 hr after LPS stimulation still enhanced IL-1ra synthesis . In human peripheral blood mononuclear cells stimulated with IL-1 alpha, IL-4 markedly suppressed IL-1 beta production but enhanced IL-1ra synthesis greater than 2-fold . Because IL-4 favors synthesis of the natural antagonist IL-1ra over synthesis of the agonist IL-1, IL-4 may exert potent antiinflammatory effects on host responses to Gram-negative infections.

J Clin Invest, 1992 May, 89(5), 1551 - 7
Interleukin-1 receptor blockade improves survival and hemodynamic performance in Escherichia coli septic shock, but fails to alter host responses to sublethal endotoxemia; Fischer E et al.; The present study was undertaken to evaluate the extent to which an endogenous interleukin-1 (IL-1) response contributes to the hemodynamic and metabolic consequences of sublethal endotoxemia or lethal Gram-negative septic shock . Young, healthy baboons received either a sublethal dose of lipopolysaccharide (LPS) or an LD100 of live Escherichia coli bacteria, and one half of the animals in each group were continuously infused with IL-1 receptor antagonist (IL-1ra) . Plasma IL-1 beta was not detected in this model of endotoxemia . Administration of IL-1ra had only minimal effects on the modest hemodynamic and metabolic responses to sublethal endotoxemia, and did not attenuate the plasma cytokine response . In contrast, high circulating levels of IL-1 beta (range 300-800 pg/ml) were seen during lethal E . coli septic shock . IL-1ra treatment significantly attenuated the decrease in mean arterial blood pressure (MAP) (from -72 +/- 8 to -43 +/- 6 mm Hg; P less than 0.05) and cardiac output (from -0.81 +/- 0.17 to -0.48 +/- 0.15 liter/min; P less than 0.05), and significantly improved survival from 43 to 100% at 24 h (P less than 0.05) . The plasma IL-1 beta and IL-6 responses to lethal E . coli septic shock were also significantly diminished by IL-1ra treatment (P less than 0.05), whereas tumor necrosis factor-alpha (TNF alpha) concentrations were unaffected . We conclude that an exaggerated systemic IL-1 beta response is characteristic of lethal E . coli septic shock, and contributes significantly to the hemodynamic and metabolic consequences of E . coli septic shock . IL-1ra can significantly attenuate the cytokine cascade and improve survival.

Am J Vet Res, 1992 May, 53(5), 748 - 52
Association between serum cytotoxicity and selected clinical variables in 240 horses admitted to a veterinary hospital; MacKay RJ; A total of 378 serum samples from 240 hospitalized horses and 47 sera from healthy control horses were assayed for growth effects on actinomycin D-treated L929 cells . On average, patient and control sera stimulated cell growth; however, mean percentage of the relative growth index (RGI) of sera from clinical cases was significantly (P less than 0.001) lower than that of control sera . Approximately 35% of patient sera and 6% of control sera had tumor necrosis factor-like cytotoxic activity for L929 cells (ie, RGI less than 100%) . Sera from horses with either peritoneal leakage of gastrointestinal tract contents or any bacterial infection were significantly (P less than 0.05) more cytotoxic than sera from horses that did not have these clinical factors . A clear tendency was evident for horses that had the highest serum cytotoxicity (RGI less than 75%) to also have clinical profiles suggestive of endotoxemia . Fever, leukopenia, diarrhea, and gastrointestinal tract leakage were significantly (P less than 0.05) overrepresented among these horses, compared with horses without serum cytotoxicity . Bacterial infections and abdominal surgeries were also increased in this group, but not significantly . Of the 14 horses with serum RGI less than 75%, 13 had some form of gastrointestinal tract disease and the other had gram-negative septicemia . Survival to discharge was significantly (P less than 0.05) lower among horses in the high-cytotoxicity group than among horses without serum cytotoxicity . Diarrhea and bacterial infections were the only clinical factors found more frequently in horses with low serum cytotoxicity than in horses without serum cytotoxicity.(ABSTRACT TRUNCATED AT 250 WORDS)

Am J Vet Res, 1992 May, 53(5), 803 - 7
Neutrophil activation associated with increased neutrophil acyloxyacyl hydrolase activity during inflammation in cattle; McDermott C et al.; Acyloxyacyl hydrolase (AOAH) is a lysosomal enzyme found in neutrophils and macrophages that acts to partially deacylate the lipid-A component of the endotoxin of gram-negative bacteria rendering it less toxic, yet maintaining much of its immunostimulatory potential . We have found that the activity of neutrophil AOAH per cell increased during localized inflammation . The purpose of this study was to determine the mechanism(s) responsible for these increases in neutrophil AOAH activity . Because changes in neutrophil maturity commonly are associated with inflammation, intravascular infusion of purified gram-negative bacterial lipopolysaccharide and SC injection of bovine recombinant granulocyte colony-stimulating factor was used to induce large numbers of circulating immature neutrophils . Immature neutrophils were found to have AOAH activity equal to that of mature cells; however, when neutrophils were stimulated in vitro with known activators, AOAH activity of activated cells was more than that of unstimulated cells . The increase in AOAH activity was inversely related to prestimulation activity . Increases in AOAH activity after neutrophil activation were not a result of de novo synthesis of the enzyme, because cycloheximide did not prevent activation-induced increases in activity.

Eur Respir J, 1992 May, 5(5), 566 - 75
Secretory leucocyte proteinase inhibitor: inhibition of fibronectin degradation by neutrophil elastase; Gervaix A et al.; Degradation of surface-bound fibronectin of the upper respiratory tract by human leucocyte elastase (HLE) was shown to favour colonization of these mucosal surfaces by Gram-negative bacteria . We investigated the degradation of fibronectin by purified HLE and by enzymes released from stimulated human polymorphonuclear leucocytes (PMNs), in the presence of recombinant secretory leucocyte proteinase inhibitor (rSLPI) and alpha 1-proteinase inhibitor (alpha 1-PI), the two main inhibitors of HLE within the airways . Our results show that HLE degraded fibronectin at concentrations as low as 0.2 nM . To inhibit the degradation of fibronectin by pure HLE in an experimental system in which the enzyme was premixed with inhibitor, a twofold molar excess of rSLPI and an equimolar concentration of alpha 1-PI were required . On the other hand, a fivefold molar excess of rSLPI was necessary to inhibit degradation of fibronectin by enzymes released from stimulated neutrophils . In order to estimate the role of oxidants generated by stimulated PMNs in the activation of the inhibitory capacity of rSLPI by stimulated PMNs, we preincubated PMNs with antioxidants such as superoxide dismutase, methionine, catalase or Na-azide prior to stimulation of the cells . Under these conditions, a threefold molar excess of rSLPI over released HLE was required to inhibit the degradation of fibronectin, raising the possibility that either exogenous or endogenous antioxidants in the lung could be important in improving the efficacy of this therapeutic antiprotease . We conclude that a molar excess of rSLPI to HLE is always necessary to inhibit fibronectin degradation by HLE, and that addition of antioxidants partly prevents the inactivation of rSLPI by oxidants released from stimulated PMNs.

J Hosp Infect, 1992 May, 21(1), 73 - 7
A randomized prospective study to compare cefotetan with cefuroxime plus metronidazole as prophylaxis in elective colorectal surgery; Skipper D et al.; This study compares the efficacy of cefotetan with the combination of cefuroxime plus metronidazole as antibiotic prophylaxis in elective colorectal surgery when given over the first 24 h postoperatively . There was no significant difference in wound infection rates between the two groups (14.7% for cefotetan and 13.9% for cefuroxime plus metronidazole), or the rates of other infective complications . Adverse reactions occurred with equal frequency in both treatment groups and no serious side effects occurred . Cefotetan is a safe and effective antibiotic for use as prophylaxis in elective colorectal surgery . Its advantages are that it is a single agent with a spectrum covering both aerobic Gram-negative rods and anaerobic organisms and, because of its long half-life, needs only to be given at 12-hourly intervals.

Gene, 1992 May 1, 114(1), 103 - 7
A simple and efficient system for the construction of phoA gene fusions in gram-negative bacteria; Duchene AM et al.; We have developed a two-plasmid system for generating gene fusions between phoA and cloned genes encoding envelope proteins . The vector plasmid carries a temperature-sensitive replication system and can be rescued at high temperature by insertion of an IS1-based transposon carrying the ori region of pBR322 and a phoA gene lacking transcription and translation initiation signals . The vector plasmid also carries the transfer origin of the conjugative plasmid, F, permitting transfer into a suitable recipient strain . We have used this system in the analysis of the bla gene cloned from pBR322.

Am J Med, 1992 Apr 6, 92(4A), 103S - 107S
Lomefloxacin versus amoxicillin in the treatment of acute exacerbations of chronic bronchitis: an Italian multicenter study; Grassi C et al.; Nine centers in Italy participated in a worldwide, multicenter study comparing the effectiveness and safety of lomefloxacin and amoxicillin in patients with acute exacerbations of chronic bronchitis caused mainly by gram-negative pathogens . The 157 enrolled patients received either 400 mg lomefloxacin once daily (n = 78) or 500 mg amoxicillin every 8 hours (n = 79) for 7-10 days . A total of 131 patients were evaluable for bacteriologic efficacy and 154 for clinical efficacy . At 2-4 days after the conclusion of treatment, the bacterial eradication rate was 84.8% for lomefloxacin-treated patients and 64.6% for amoxicillin-treated patients (p = 0.0065); the clinical success rate (cure plus improvement) for lomefloxacin was 94.7% and for amoxicillin was 83.3% (p = 0.0212) . The reinfection rate was lower in the lomefloxacin group than in the amoxicillin group (3.0% vs 13.8%, p = 0.0382) . Both drug regimens were well tolerated . Once-a-day treatment with 400 mg lomefloxacin was more effective than 500 mg amoxicillin three times daily for the treatment of acute exacerbations of chronic bronchitis caused by gram-negative pathogens.

Pathol Res Pract, 1992 Apr, 188(3), 314 - 7
Ultrastructure of Warthin-Starry stain-positive bacteria in abscess-forming reticular lymphadenitis; Ohtani Y et al.; This study describes the ultrastructure of Warthin-Starry (WS) stain-positive bacteria in abscess-forming reticular lymphadenitis (ARL) compatible with cat scratch disease (CSD) . Sections containing WS-positive bacteria were re-embedded in Epon, and semithin sections were examined by electron microscopy . Silver particles were aggregated on the outer surfaces of the bacteria . Stereoscopic observations clearly showed that the bacteria were pleomorphic, rod-shaped, arranged in a row or at an angle, and frequently showed septal formation . Electron microscopy of ultrathin sections revealed that the cell wall possessed an outer membrane characteristic of gram-negative bacteria . The results indicate that rod-shaped bacteria with the WS-positive, gram-negative staining cause ARL histopathologically consistent with CSD.

J Invest Surg, 1992 Apr-Jun, 5(2), 129 - 41
Use of myxalin for improving vascular graft healing: evaluation of biocompatibility in rats; Akoum A et al.; Myxalin is a glycopeptide extracted recently from a gram-negative bacterium . It has blood anticoagulant properties and can enhance endothelial cell growth . With the ultimate objective of using this bioactive molecule to promote vascular graft healing, this study assessed its biocompatibility in vivo by comparing the cellular and immunological responses of gelatin-coated knitted polyester grafts with and without myxalin following implantation in the peritoneal cavity of rats for prescheduled periods of 3 days and 1, 2, and 4 weeks . A nongelatin-coated virgin polyester graft was included as the reference material . The biological response to gelatin alone was characterized by a slower rate of cellular infiltration into the implant, reduced collagen synthesis, and higher levels of acid phosphatase and esterase activity in the surrounding tissue . The addition of myxalin to this coating resulted in a significant reduction of hydrolase secretion in the tissue surrounding the implant and an enhancement of cellular ingrowth.

J Surg Res, 1992 Apr, 52(4), 314 - 9
Protective anti-lipopolysaccharide monoclonal antibodies inhibit tumor necrosis factor production; Cody CS et al.; Elevated systemic levels of tumor necrosis factor (TNF) have been directly correlated with increased mortality during experimental gram-negative bacterial sepsis . Although monoclonal antibodies (mAbs) directed against gram-negative bacterial lipopolysaccharide (endotoxin, LPS) decrease TNF production in vitro and enhance survival in vivo, the precise relationship between inhibition of TNF secretion and protective capacity has not been defined . We hypothesized that protective anti-LPS mAbs inhibited LPS-stimulated TNF production . To test this hypothesis, we first produced and characterized three anti-LPS mAbs . We then examined the ability of these mAbs to decrease TNF secretion in an in vitro assay using cells from the murine macrophage cell line RAW 264.7 . Subsequently, we assessed the protective capacities of these anti-LPS mAbs in a murine mucin peritonitis model of sepsis using live Escherichia coli 0111:B4 bacterial challenge . Our results demonstrated that those anti-LPS mAbs that decreased LPS-stimulated TNF secretion in vitro were protective in vivo . We concluded that inhibition of TNF secretion in vitro reflected protective capacity and that anti-LPS mAbs may confer protection via abrogation of macrophage TNF secretion . Inhibition of TNF production in vitro may provide a valuable test that may facilitate the selection of protective anti-LPS mAbs.

Br J Urol, 1992 Apr, 69(4), 372 - 4
Ethics, logistics and a trial of transurethral versus open prostatectomy; Jenkins BJ et al.; An increased long-term morbidity rate after transurethral compared with open prostatectomy has recently been claimed on the basis of retrospective studies of operations done up to 20 years ago . These studies have led to a demand for a prospective trial . Most reports show that peri-operative mortality following prostatectomy is virtually confined to unfit men over 80 years old . Before agreeing to participate in a trial we examined our operative mortality in this group of patients . Between 1981 and 1987, 123 octogenarians underwent transurethral prostatectomy: 64 operations were elective and 59 were performed for retention . There were 2 operative deaths (1.6%), both from gram-negative septicaemia despite prophylactic antibiotics . There were no additional deaths in the first 12 months following surgery . Few of these patients would have been considered fit to undergo an open prostatectomy . Any proposed randomised trial would have to exclude such high risk patients until it can be shown that open prostatectomy is equally safe.

Br J Surg, 1992 Apr, 79(4), 300 - 4
Role of selective decontamination in surgery; Tetteroo GW et al.; Aspects of selective decontamination as an antibiotic prophylaxis in general surgery are discussed with respect to the respiratory tract and wound infection . The high rate of nosocomial infection encountered in surgical patients, who experience up to 71 per cent of all such infection, is considered . The infection rate increases to 70-80 per cent after 1 week in the surgical intensive care unit, while the infection-related mortality rate rises to up to 60 per cent . The endogenous origin of nosocomial infection suggests the importance of maintaining colonization resistance to prevent colonization with Gram-negative micro-organisms, especially using selective decontamination which eliminates potentially pathogenic micro-organisms while leaving colonization resistance intact . Selective decontamination has proved effective in decreasing infection rates by 50-80 per cent, and is especially successful in respiratory tract infection . Possible adverse effects and future indications for selective decontamination are discussed.

Clin Infect Dis, 1992 Apr, 14(4), 908 - 15
Alterations in the microbial flora and in the incidence of bacteremia at a university hospital after adoption of amikacin as the sole formulary aminoglycoside; King JW et al.; Because of the rapid emergence of resistance to gentamicin and tobramycin among isolates of aerobic and facultative gram-negative bacteria at our university hospital, we designed a prospective study to track aminoglycoside resistance, bacteremic episodes, and bacteremia-associated deaths before and after the institution of amikacin as the sole formulary aminoglycoside . From June 1984 through June 1987 (immediately before this policy change), amikacin accounted for only 20% of patient-days of aminoglycoside therapy, and rates of resistance to gentamicin, tobramycin, and amikacin among aerobic and facultative gram-negative bacterial isolates were 12.8%, 10.8%, and 5.9%, respectively . During the next 30 months (immediately after the change in policy), amikacin accounted for 98% of patient-days of aminoglycoside therapy, and rates of resistance to gentamicin, tobramycin, and amikacin were 6.3%, 5.0%, and 3.3%, respectively . Furthermore, during the latter 30 months, the incidence of both bacteremia and bacteremia-associated death decreased significantly . Hospitals at which resistance to gentamicin or tobramycin is increasing among the gram-negative flora may benefit from the use of amikacin as the principal aminoglycoside.

J Mol Evol, 1992 Apr, 34(4), 358 - 75
Phylogenetic continuum indicates "galaxies" in the protein universe: preliminary results on the natural group structures of proteins; Ladunga I; The markedly nonuniform, even systematic distribution of sequences in the protein "universe" has been analyzed by methods of protein taxonomy . Mapping of the natural hierarchical system of proteins has revealed some dense cores, i.e., well-defined clusterings of proteins that seem to be natural structural groupings, possibly seeds for a future protein taxonomy . The aim was not to force proteins into more or less man-made categories by discriminant analysis, but to find structurally similar groups, possibly of common evolutionary origin . Single-valued distance measures between pairs of superfamilies from the Protein Identification Resource were defined by two chi 2-like methods on tripeptide frequencies and the variable-length subsequence identity method derived from dot-matrix comparisons . Distance matrices were processed by several methods of cluster analysis to detect phylogenetic continuum between highly divergent proteins . Only well-defined clusters characterized by relatively unique structural, intracellular environmental, organismal, and functional attribute states were selected as major protein groups, including subsets of viral and Escherichia coli proteins, hormones, inhibitors, plant, ribosomal, serum and structural proteins, amino acid synthases, and clusters dominated by certain oxidoreductases and apolar and DNA-associated enzymes . The limited repertoire of functional patterns due to small genome size, the high rate of recombination, specific features of the bacterial membranes, or of the virus cycle canalize certain proteins of viruses and Gram-negative bacteria, respectively, to organismal groups.

Proc Natl Acad Sci U S A, 1992 Apr 1, 89(7), 2713 - 7
Transport and anchoring of beta-lactamase to the external surface of Escherichia coli; Francisco JA et al.; The outer membrane of Gram-negative bacteria presents an effective barrier that restricts the release of proteins from the cell . Virtually all extracellular proteins of Gram-negative bacteria are exported by specialized systems requiring the action of several gene products . We have constructed a tripartite fusion consisting of (i) the signal sequence and first nine N-terminal amino acids of the mature major Escherichia coli lipoprotein, (ii) amino acids 46-159 of the outer membrane protein OmpA, and (iii) the complete mature beta-lactamase (EC 3.5.2.6) sequence . This protein had an enzymatically active beta-lactamase and was found predominantly in the outer membrane . Immunofluorescence microscopy, the accessibility of the fusion protein to externally added proteases, and the rates of hydrolysis of nitrocefin and penicillin G by whole cells demonstrated that a substantial fraction (20-30%) of the beta-lactamase domain of the fusion protein was exposed on the external surface of E . coli . In cells grown at 24 degrees C the localization of beta-lactamase on the cell surface was almost quantitative (greater than 80% of the enzymatically active protein was exposed to the extracellular fluid) as determined by nitrocefin and penicillin G hydrolysis and trypsin accessibility . These results demonstrated that a soluble protein, beta-lactamase, can be transported through--and become anchored on--the outer membrane by fusion to the proper targeting and localization signals.

Chest, 1992 Apr, 101(4), 1056 - 9
Infectious complications following isolated lung transplantation; Maurer JR et al.; STUDY OBJECTIVE: To ascertain the incidence, types, morbidity, and mortality of infectious episodes in isolated lung transplant recipients . DESIGN: Retrospective chart review of patients who have undergone transplants over a six-year period in one institution . PATIENTS: Twenty-three single and 17 double lung transplants followed up between 2 and 68 months . RESULTS: Fifty-one episodes of infection occurred in the group with a slight predominance in the double lung transplants . The 32 episodes of bacterial infection constituted the largest group of infection and more than half of these were pneumonias . Organisms identified were predominantly Gram negative . While bacterial processes made up the bulk of infections, fatalities were rare . Viral and fungal infections were less common, but more often fatal . Of six cases of viral pneumonitis, two were fatal; two of five cases of invasive fungal infection were also fatal . Overall, six patients died of infection . CONCLUSION: Our findings support previous reports from heart-lung centers documenting a high rate of infectious complications, particularly pneumonia, in recipients of lung grafts . In our experience, bacterial infections are the most common (two of three infections), but have the lowest mortality . Efforts should be directed toward establishing effective prophylaxis programs and early detection of infection.

J Infect Dis, 1992 Apr, 165(4), 683 - 8
Nephrotoxicity and ototoxicity of aztreonam versus aminoglycoside therapy in seriously ill nonneutropenic patients; Moore RD et al.; A randomized double-blind clinical trial was done of aztreonam versus aminoglycoside therapy for the empiric treatment of seriously ill nonneutropenic patients suspected of aerobic gram-negative bacterial infection . Each patient was treated for greater than or equal to 72 h with the study drug . Nephrotoxicity, defined by greater than or equal to 50% increase in baseline serum creatinine, occurred in 12 (15%) of 92 patients receiving aminoglycoside therapy and 1 (1%) of 92 patients receiving aztreonam (P less than .004) . More severe nephrotoxicity, defined by greater than or equal to 100% increase in baseline serum creatinine, occurred in 6 (6.5%) of 92 patients receiving aminoglycoside therapy and in 1 of 92 receiving aztreonam (P less than .11) . Patients with an elevated baseline total bilirubin level were most likely to develop nephrotoxicity . Auditory toxicity occurred in 2 (7%) of 28 evaluatable patients receiving aminoglycoside therapy and in 1 (3%) of 33 receiving aztreonam (P less than .58) . One patient, who received aminoglycoside, developed vestibular toxicity . In nonneutropenic patients believed to be at increased risk for renal dysfunction, aztreonam is a less toxic alternative to aminoglycoside therapy for treatment of suspected aerobic gram-negative infection.

Hinyokika Kiyo, 1992 Apr, 38(4), 507 - 10
{Clinical study of cefclidin on bacterial prostatitis}; Suzuki K et al.; Cefclidin (CFCL), a new injectable cephem antibiotic, was used in the treatment of bacterial prostatitis . Concentration in prostatic fluid (PF): One hour after the i.v . administration of 1 g of CFCL the concentration of PF was 0.84 +/- 0.72 micrograms/ml (n = 4), the ratio of PF/serum being 2.24% . CFCL was administered at a dose of 1-2 g a day for 6.5 days on average . In acute bacterial prostatitis, the efficacy was evaluated as excellent or moderate in all 7 cases (100%), but in chronic cases, the effectiveness rate was as low as one out of 4 cases (25%) . In safety profile, transient skin eruption was seen in one patient, but did not require further treatment . Besides this case, no side reactions or abnormal clinical values were encountered . In conclusion, CFCL was evaluated to be very useful in the treatment of acute bacterial prostatitis, caused by gram negative rods.

Bratisl Lek Listy, 1992 Apr, 93(4), 175 - 8
{Bacterial resistance to aminoglycoside antibiotics}; Kettner M et al.; Aminoglycoside antibiotics, such as gentamicin, netilmicin, tobramycin, amikacin, may be inactivated by resistant bacteria producing three types of enzymes: O-nucleotidyltransferases, O-phosphotransferases and N-acetyltransferases . Mechanisms of aminoglycoside resistance were analyzed in clinical isolates of gram-negative bacteria from several regions of Czecho-Slovakia and compared with the use of aminoglycosides . Production of acetyltransferases (AAC) and nucleotidyltransferases (ANT) was observed in 84% of bacterial isolates . The majority of bacterial strains studied produced the AAC/3/enzyme (62%) . Due to dissemination of plasmids coding for the AAC/3/enzyme, the majority of Czecho-Slovak gentamicin-resistant strains was also tobramycin resistant (93%) and netilmicin resistant (68%) . Amikacin remains the most effective aminoglycoside antibiotic against multiresistant bacterial strains.

J Infect Dis, 1992 Apr, 165(4), 658 - 66
Characterization of antigenic determinants of Borrelia burgdorferi shared by other bacteria; Coleman JL et al.; Three antigenic determinants of Borrelia burgdorferi that were shared with other spirochetes and gram-negative bacterial species, as measured by Western blot, ELISA, or both, were identified and characterized using monoclonal antibodies (MAbs) . Two were associated with immunogenic integral membrane lipoproteins of 19- and 22-kDa, respectively, by {3H}palmitate incorporation and Triton X-114 phase partitioning . A third antigenic determinant was shown to be associated with a 72-kDa heat shock protein that was also immunogenic in human patients . MAb agglutination assays with B . burgdorferi and treatment of the spirochete with proteases indicated that none of the antigenic determinants were surface exposed . NH2-terminal sequence analysis revealed the 72-kDa protein to have 100% identity with the first 13 amino acid residues of the Escherichia coli dnaK heat shock protein . The presence of these and other shared antigenic determinants in ELISA antigen preparations could explain the high degree of serologic cross-reactivity in current diagnostic procedures.

Microbiologica, 1992 Apr, 15(2), 187 - 90
The presence in human spermatozoa of sites binding some proteins of the outer membrane of Escherichia coli; Gorga F et al.; In previous works, we noted that porins, surface components of Gram negative bacteria, are toxic for human spermatozoa . In this work we see that porins of E . coli labeled with I125 (0.1-1.0 microgram/ml) bind to spermatozoa, showing the presence of saturable molecular configurations of 0.5-0.6 micrograms/ml . In the presence of non labeled porins, the binding of labeled porins to the spermatozoa is greatly reduced . At saturating concentrations of 0.6 micrograms/ml labeled porins, the binding reaches a saturation plateau at about 15 min.

Biochim Biophys Acta, 1992 Mar 25, 1124(3), 233 - 40
Disturbances in the composition of plasma lipoproteins during gram-negative sepsis in the rat; Lanza-Jacoby S et al.; Hyperlipidemia is associated with gram-negative sepsis . In this study we characterized the plasma lipoproteins of fasted and fed septic and control rats with respect to their lipid and apolipoprotein composition . Sepsis was induced by i.v . injection of 8 x 10(7) live Escherichia coli colonies/100 g body wt . Food was removed from fasted control and fasted E . coli-treated rats after injection . Fed rats were infused intragastrically with a nutritionally complete diet for 5 days prior to E . coli treatment . 24 h after treatment with E . coli, lipid and protein concentrations of very-low-density lipoprotein (VLDL) were over 2-fold higher in the fasted E . coli-treated rats than those of the fasted control rats . This appears to be due to a decrease in the clearance of VLDL . The relative composition of apolipoprotein B-48 and apolipoprotein E were lower while that of apolipoprotein B-100 was higher in fasted E . coli-treated rats than in fasted controls . Low-density lipoprotein (LDL) and high-density lipoprotein lipids were also significantly elevated, indicating greater synthesis of these particles during sepsis and food deprivation . By contrast, VLDL-triacyglycerol from fed, E . coli-treated did not differ from that of their respective controls although the total cholesterol remained elevated . Percentages of apolipoprotein B-48 and apolipoprotein B-100 increased while apolipoprotein E contributed significantly less to the total protein of VLDL from the E . coli-treated rats compared with controls . LDL lipids were also increased . In conclusion, gram-negative sepsis leads to marked changes in the plasma lipoprotein composition which may be attributed to altered hepatic synthesis, peripheral metabolism or hepatic uptake of lipoproteins and their remnants . These in turn may be a function of the nutritional status.

J Immunol, 1992 Mar 15, 148(6), 1890 - 7
Cytokine production after intravenous or peritoneal gram-negative bacterial challenge in mice . Comparative protective efficacy of antibodies to tumor necrosis factor-alpha and to lipopolysaccharide; Zanetti G et al.; The production of TNF-alpha, IL-1, and IL-6 was measured in mice after bolus i.v . Escherichia coli O111 LPS injections and during bacteremia induced either by bolus i.v . or by i.p . challenges of live E . coli O111 . High but transient TNF-alpha peaks were observed after bolus i.v . LPS or bacterial challenges . In contrast, the levels during lethal peritonitis increased progressively to values 50- to 100-fold lower than the peak values observed after i.v . injections, and remained sustained until death . Whereas after i.v . challenge with 1000 LD50 of LPS, anti-TNF-alpha antibody fully protected mice from death and reduced serum IL-1 and IL-6 levels, anti-TNF-alpha antibody did not improve the survival of mice nor reduced serum IL-1 and IL-6 levels after i.p . bacterial challenge . In contrast to anti-TNF-alpha antibodies, anti-LPS antibodies were protective in the peritonitis model . Protection was accompanied by a striking reduction of bacterial numbers and of TNF-alpha, IL-1, and IL-6 levels in the serum, but the levels of these cytokines were only marginally affected in the peritoneal lavage fluid . This latter observation demonstrates that the local peritoneal cytokines did not diffuse readily into the circulation, thus suggesting that at least part of the circulating cytokines are produced systemically . In conclusion, the striking differences between cytokine profiles as well as the divergent efficacy of anti-TNF-alpha antibody after i.v . bolus and after i.p . challenges suggest that TNF-alpha may not be as important in the pathogenesis of lethal peritonitis than after lethal acute bacteremia.

Eur J Clin Microbiol Infect Dis, 1992 Mar, 11(3), 237 - 40
Moraxella catarrhalis bacteremic pneumonia in adults: two cases and review of the literature; Collazos J et al.; Moraxella (formerly Branhamella) catarrhalis is a gram-negative coccus now recognized as one of the common pathogens in respiratory infections . Documented cases of bacteremic pneumonia due to this organism, however, have been a rarity . Two cases of Moraxella catarrhalis bacteremic pneumonia in immunosuppressed adult patients are reported . The clinical characteristics of these patients together with those of the seven adult and the six pediatric patients reported to date in the literature, are analyzed . All patients had an underlying condition and most were male . The mean age was 64.9 years . No adult patient had skin lesion, although purpuric rash was frequent in children . The overall morality rate was only 13.3%, in spite of the underlying diseases . In three patients the pneumonia was nosocomial . The seasonal recovery of Moraxella catarrhalis in respiratory infections is significantly increased during the late fall through early spring period . Because most strains are beta-lactamase positive, empiric use of penicillin, ampicillin or amoxicillin for this organism can no longer be recommended.

Clin Exp Allergy, 1992 Mar, 22(3), 417 - 9
Type I allergy induced by Limulus Amoebocyte Lysate (LAL); Ebner C et al.; During the last few years Limulus Amoebocyte Lysate (LAL) has been extensively used to detect minimal amounts of endotoxins of Gram-negative bacteria in products of the pharmaceutical industry, in food stuff, body fluids, house dust and room air . LAL is produced from cells of the haemolymph (amoebocytes) of the horseshoe crab (Limulus polyphemus), which respond with an extremely sensitive clotting system upon contact with endotoxins . In this study we demonstrate by typical case history, positive skin test and ELISA the occurrence of Type I allergy to LAL in a patient suffering from conjunctivitis and rhinitis at work.

Infect Dis Clin North Am, 1992 Mar, 6(1), 253 - 66
HA-1A . A human monoclonal antibody for the treatment of gram-negative sepsis; Smith CR et al.; HA-1A is a human monoclonal IgM antibody that binds to endotoxin . The results of the clinical trials of HA-1A demonstrate that HA-1A reduces mortality among patients with sepsis and gram-negative bacteremia . Secondary endpoints, including resolution of organ failure, discharge from intensive care unit, and discharge from the hospital, support the beneficial effects of the antibody . The antibody is well tolerated with rare side effects, including hypotension and urticarial rash . No anti-HA-1A antibodies have been detected.

Am J Physiol, 1992 Mar, 262(3 Pt 2), H710 - 8
EDRF and nitric oxide production in cultured endothelial cells: direct inhibition by E . coli endotoxin; Myers PR et al.; Recent studies have yielded contradictory interpretations about the influence of gram-negative endotoxin on endothelium-derived relaxing factor (EDRF) . We tested the hypothesis that Escherichia coli endotoxin exerts primary facilitatory or, alternatively, inhibitory actions on EDRF release and the synthesis of either nitric oxide or a nitroso compound in cultured endothelial cells . Bovine aortic endothelial cells were grown on microcarrier beads and either exposed acutely (30 min) to E . coli endotoxin or incubated with endotoxin for 1 h followed by a 1-h wash (prolonged exposure) . EDRF bioactivity was measured under basal, bradykinin-stimulated, and A23187-stimulated conditions using standard isometric tension recordings . EDRF-derived nitric oxide was quantitated using a specific chemiluminescence technique . Endotoxin (0.005-5 micrograms/ml) decreased EDRF bioactivity and nitric oxide production under both basal and bradykinin-stimulated conditions after prolonged, but not acute, exposure . A23187-stimulated EDRF bioactivity and nitric oxide production were minimally, albeit significantly, reduced after endotoxin . The present results demonstrate that EDRF activity and nitric oxide production are decreased in vascular endothelial cells exposed to endotoxin . Endotoxin itself failed to directly stimulate EDRF release from endothelium . Alternative sources of nitrovasodilators, endothelium-independent effects, or release of other vasoactive mediators by endotoxin may be responsible for systemic hypotension during in vivo endotoxemia.

Arch Intern Med, 1992 Mar, 152(3), 529 - 35
Predictors of bacteremia and gram-negative bacteremia in patients with sepsis . The Veterans Affairs Systemic Sepsis Cooperative Study Group; Peduzzi P et al.; BACKGROUND--We analyzed data from the Department of Veterans Affairs trial of steroid therapy for systemic sepsis to identify predictors of bacteremia and gram-negative bacteremia . METHODS--Of the 2568 patients screened for entry in the trial, 465 met the following criteria: presence of four of seven clinical signs of sepsis; blood cultures at the time of screening; and complete data on nine clinical parameters . The multivariate logistic regression model was used to identify predictors of bacteremia and gram-negative bacteremia . Predicted probabilities of having these types of infections were calculated using the identified predictors . Patients were then classified into groups with and without bacteremia (and gram-negative bacteremia) based on the predicted probability . Misclassification error rates were calculated for each method of categorization by comparing the true with the predicted grouping of patients . RESULTS--Three factors were independently predictive of bacteremia and gram-negative bacteremia: elevated temperature, low systolic blood pressure, and low platelet count . Using these three factors, classification methods were identified that predicted blood infection better than chance, but misclassification was also high . For predicting bacteremia, the maximum predicted positive rate was 83%, with a specificity of nearly 100% and a sensitivity of only 5% . For predicting gram-negative bacteremia, the maximum predicted positive accuracy was 100%, with a specificity also of 100% and a sensitivity of almost 0% . CONCLUSIONS--Using simple clinical parameters, we could not predict either bacteremia or gram-negative bacteremia with sufficient accuracy to be clinically meaningful; however, our approach represents a step in the direction of forecasting the bacterial organism responsible for sepsis in advance of culture results.

Surg Gynecol Obstet, 1992 Mar, 174(3), 225 - 8
Repeat median sternotomy for the automatic implantable cardioverter and defibrillator; Blakeman BP et al.; Forty-four patients with a repeat sternotomy for the implantation of the automatic implantable cardioverter and defibrillator are presented . Thirty-three of the patients had placement of the device only and 11 had a concomitant open heart procedure--nine for aorto-coronary bypass (mean of 1.6) and two for mitral valve replacement . Twenty-two complications occurred in 17 patients . Arrhythmia was the most common problem . Two deaths occurred perioperatively in the group (mortality rate of 4.5 per cent) and were caused by gram-negative pneumonia and pulmonary embolus . There were no specific complications related to the sternotomy . It is, therefore, concluded that the repeated sternotomy is an excellent method to insert the automatic cardioverter and defibrillator.

Bratisl Lek Listy, 1992 Mar, 93(3), 131 - 5
{Mechanisms of resistance to dactimicin and isepamicin in clinical strains of gram-negative rods isolated in Czechoslovakia}; Kettner M et al.; Dactamicin is a new member of the pseudo-disaccharide group of antibiotics . It is structurally close to astromicin . Dactimicin is active against a wide variety of bacteria, including resistant strains with aminoglycoside-inactivating enzymes . Isepamicin is a broad-spectrum aminoglycoside, a derivative of gentamicin B . It is also active against many aminoglycoside-resistant strains . In a series of gram-negative bacterial isolates from various Czecho-Slovak regions, in vitro investigation yielded a good activity of dactimicin and isepamicin against amikacin- and gentamicin-resistant strains producing the enzyme AAC/6'/ or a combination of AAC/6'/ + ANT/2"/ enzymes . Dactimicin was not effective against strains producing the enzyme AAC/3/-I, and isepamicin was not effective or only slightly effective against bacterial isolates producing a combination of AAC/6'/ + AAC/3/ enzymes . (Tab . 3, Fig . 2, Ref . 10.).

Curr Opin Dent, 1992 Mar, 2, 109 - 14
Treatment of failing dental implants; Meffert RM; Recent studies show that the bacterial flora in adult periodontitis and peri-implantitis are identical, but there could be a more pathogenic flora (gram-negative bacteria, spirochetes) in the partially edentulous implant case than in the fully edentulous implant case . These data indicate a possible seeding mechanism from the natural tooth pocket to the implant crevice and may place the implant in the partially edentulous case more at risk . Also, understanding of the importance of loading in a progressive sense to prevent retrograde peri-implantitis involving the dental implant has increased; the density of bone must be evaluated more fully to determine the time frame necessary to gain a compact bone capable of withstanding occlusal forces at the bone-implant interface.

Minerva Pediatr, 1992 Mar, 44(3), 79 - 86
{The use of ofloxacin in cystic fibrosis patients}; Romano L et al.; The new quinolones represent the latest possibility of specific oral antibiotic treatment of infections caused by gram-negative bacteria . Among the new quinolones, ciprofloxacin and ofloxacin are characterized by strong in vitro activity against most Pseudomonas species strains, favourable kinetic in body fluids, good tolerability and the possibility of oral administration . For these reasons they appear to be ideal antibiotics for long-term home therapy of chronic obstructive pulmonary disease in cystic fibrosis (CF) . The efficacy of ciprofloxacin has been recently assessed . In this study, actual effectiveness of ofloxacin in long-term home antibiotic treatment of patients affected by CF was evaluated . The study was a no-blind cross-over study, designed to compare ofloxacin treatment with conventional oral antibiotic therapy . Young adult patients, who needed long-term antibiotic therapy and whom sputum culture were positive for sensitive strains, were randomly assigned to 2 groups . One group received ofloxacin, the other group was given a non-quinolone oral antibiotic, selected according to sputum culture sensitivity . Oral antibiotics were administered for 20 days, then a break of 10 days was allowed during which patients received nebulized aminoglucosides, usually tobramycin . After 3 months, therapies were rotated: the first group received a non-quinolone oral antibiotic and the second group received ofloxacin for another 3 months . The clinical score (according to Huang et al., see table I) and the lung function (FVC, FEV1, pulsed SaO2) were assessed in all the patients at the beginning and at the end of each three months period of oral antibiotic therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

FEBS Lett, 1992 Feb 24, 298(2-3), 273 - 6
Identification and solubilization of a signal peptidase from the phototrophic bacterium Rhodobacter capsulatus; Wieseler B et al.; In Gram-negative bacteria, exported proteins are synthesized with an amino-terminal signal sequence which is cleaved off by the signal peptidase during, or shortly after the translocation process . Here, we report the identification and solubilization of a signal peptidase from the phototrophic bacterium Rhodobacter capsulatus which cleaves homologous and heterologous precursor proteins at the authentic cleavage site . This signal peptidase is the first identified component of the R . capsulatus protein export machinery.

J Surg Res, 1992 Feb, 52(2), 161 - 6
The effect of albumin or crystalloid resuscitation on bacterial translocation and endotoxin absorption following experimental burn injury; O'Brien R et al.; Burn injury induces immune suppression and increases susceptibility to infection . Hypoalbuminemia is an early and consistent finding following thermal injury and is independently associated with gastrointestinal dysfunction and increased rates of infectious morbidity . This study assessed the effects of albumin resuscitation on burn-induced immunosuppression, bacterial translocation, and absorption of gut endotoxin . Male Sprague-Dawley rats, presensitized to keyhole limpet hemocyanin (KLH), underwent a 20% dorsal scald burn injury, followed by laparotomy and IVC catheterization for fluid resuscitation . Animals were randomized to one of three resuscitative regimens: Ringer's lactate 3 ml/kg/% burn, Ringer's lactate 9 ml/kg/% burn, or 5% human albumin 3 ml/kg/% burn . Delayed hypersensitivity (DTH) responses to KLH were depressed 24 hr following injury (preburn 8.9 +/- 0.2 mm, post-burn 3.1 +/- 0.3 mm, P less than 0.001) and were significantly lower in animals in whom gram-negative bacterial translocation had occurred (2.3 +/- 0.4 vs 3.6 +/- 0.2 mm, P less than 0.005) . Serum albumin levels were lower and rates of gram-negative bacterial translocation higher for those animals receiving low volume crystalloid resuscitation; animals resuscitated with albumin or high volume crystalloid experienced similar degrees of postinjury hypoalbuminemia and bacterial translocation . Uptake of radiolabeled endotoxin was maximal in animals resuscitated with albumin . Bacterial translocation is believed to be responsible for a significant number of late nosocomial infections following trauma . These data suggest that the adequacy of early resuscitation rather than the type of resuscitative solution is the more important factor in minimizing translocation.

FEMS Microbiol Immunol, 1992 Feb, 4(3), 123 - 8
Partial characterization of a cell-free hemolytic factor produced by Helicobacter pylori; Wetherall BL et al.; Maximum cell-free hemolytic activity of helicobacter pylori cultured in broth containing 10% horse serum occurred only after the stationary phase of growth was reached, unlike many hemolysins produced by Gram-negative bacteria which are active during exponential growth . This characteristic of the H . pylori hemolytic factor suggested that it might also possess protease activity . However, because no evidence of albumin degradation was found, the hemolysis by cell-free concentrates of H . pylori appears to be due to a unique factor derived from the organism . Because variable hemolysis results were obtained with culture broths lacking albumin or serum, these proteins may act as carriers or stabilizers of the putative hemolysin.

FEMS Microbiol Rev, 1992 Feb, 8(2), 137 - 61
Structural and functional relationships among the RTX toxin determinants of gram-negative bacteria; Coote JG; The RTX (repeats in toxin) cytolytic toxins represent a family of important virulence factors that have disseminated widely among Gram-negative bacteria . They are characterised by a series of glycine-rich repeat units at the C-terminal end of each protein . They also have other features in common . Secretion from the cell occurs without a periplasmic intermediate by a novel mechanism which involves recognition of a signal sequence at the C-terminus of the toxin by membrane-associated proteins that export the toxin directly to the outside of the cell . The structural gene for each protein encodes an inactive toxin which is modified post-translationally to an active cytotoxic form by another gene product before secretion . The genes for toxin synthesis, activation and secretion are for the most part grouped together on the chromosome and form an operon . The toxins all create pores in the cell membrane of target cells leading to eventual cell lysis and they appear to require Ca2+ for cytotoxic activity . Although the toxins have a similar mode of action, they vary in target cell specificity . Some are cytotoxic for a wide variety of eukaryotic cell types while others exhibit precise target cell specificity and are only active against leukocytes from certain host species . The characteristic glycine-rich repeat units have been identified in other exoproteins besides those with cytotoxic activity and it is likely that the novel secretory mechanism has been harnessed by a variety of pathogens to release important virulence-associated factors from the cell or to locate them on the cell surface.

Thorax, 1992 Feb, 47(2), 115 - 8
Community acquired acute bacterial and atypical pneumonia in Saudi Arabia; Kurashi NY et al.; BACKGROUND: Rational treatment of pneumonia requires knowledge of the likely aetiological diagnosis in any community . Little is known about the pattern and outcome of pneumonia in Saudi Arabia . METHODS: A total of 567 pneumonic episodes in adult patients from the Al-Qassim area were reviewed retrospectively . RESULTS: Patients had a mean age of 42.7 years, with 103 patients (18%) aged 13 to 20 years and 103 (18%) aged 60 or more . Almost two thirds of the episodes (64%) occurred in men . An aetiological diagnosis was established in 351 (62%) cases, with 145 episodes being due to pneumococcal infection and 129 to Mycoplasma pneumonia . Inhospital mortality was 6% (35 patients) . Age over 60 years, aspiration pneumonia, and Gram negative infection were the only factors that independently predicted adverse outcome on adjusted mortality analysis . CONCLUSION: This analysis of pneumonia in the Al-Qassim area indicates the pattern and prognosis of acute bacterial and atypical pneumonia that requires admission to hospital in the central region of Saudi Arabia . It should provide a basis for developing rational treatment for community acquired pneumonia in Saudi Arabia.

Oral Microbiol Immunol, 1992 Feb, 7(1), 7 - 13
Identification of Selenomonas species by whole-genomic DNA probes, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, biochemical tests and cellular fatty acid analysis; Maiden MF et al.; Nonisotopic, whole-genomic DNA probes, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), biochemical tests in microtiter trays and cellular fatty acid (CFA) analysis were compared for the identification of 5 oral Selenomonas species . DNA probes were prepared by biotin-labeling DNA extracted from the type strains of Selenomonas noxia, Selenomonas flueggei, Selenomonas artemidis, Selenomonas infelix and Selenomonas sputigena . The probes were hybridized with DNA from 21 reference strains, 18 fresh isolates of Selenomonas species, and 21 strains of other oral gram-negative species . Target DNAs were obtained by in situ extraction of colonies blotted onto filter paper . Streptavidin-linked alkaline phosphatase was used to detect homologous reactions of probe and target DNA . Each Selenomonas species DNA probe reacted with reference strains of only that species . All Selenomonas strains that reacted with the DNA probe for a particular species gave similar biochemical test results, SDS-PAGE protein profiles, and CFA profiles to those of the type strain of the corresponding species . All the methods tested were useful for identifying the species, and all yielded similar identifications of the fresh isolates . The DNA probes, however, had the potential for identifying Selenomonas species directly from primary isolation plates or plaque samples.

J Chemother, 1992 Feb, 4(1), 27 - 9
Penetration of ciprofloxacin into human peritoneal tissue following a single oral dose of 750 milligrams; Dan M et al.; The penetration of ciprofloxacin into the peritoneal tissue was studied in 10 patients after a single oral dose of 750 mg given 5.6 hours (mean) before elective laparotomy . The mean tissue level was 0.29 micrograms/ml (range, 0.082 to 0.96 micrograms/ml) while the mean concomitant serum level was 1.3 micrograms/g (range, 0.52 to 2.57 micrograms/g) . The achieved concentrations are above the minimum inhibitory concentrations (MICs) of ciprofloxacin for most gram-negative bacteria commonly involved in intra-abdominal infections.

Eur J Clin Microbiol Infect Dis, 1992 Feb, 11(2), 162 - 3
Spleen abscess caused by Eikenella corrodens; Perez-Pomata MT et al.; A case is reported of splenic abscess due to Eikenella corrodens, a gram-negative rod which is found as part of normal flora in human mucous surfaces . A 64-year-old man presented with fever, chills, anorexia and abdominal pain . Abdominal ultrasound examination showed a perisplenic fluid collection which was considered to be either blood or a subcapsular spleen abscess . The presence of a splenic abscess was later confirmed during surgery and a splenectomy was performed . Splenic purulent material and blood cultures yielded Eikenella corrodens . The patient received cefotaxime for 19 days and was discharged asymptomatic.

J Bioenerg Biomembr, 1992 Feb, 24(1), 71 - 5
Evidence for extra-mitochondrial localization of the VDAC/porin channel in eucaryotic cells; Thinnes FP; The expression of bacterial porin in outer membranes of gram-negative bacteria and of mitochondrial porin or voltage-dependent anion channel (VDAC) in outer mitochondrial membranes (OMM) of eucaryotic cells was demonstrated about 15 years ago . However, the expression of VDAC in the plasmalemma (PLM) of transformed human B lymphoblasts has recently been indicated by cytotoxicity and indirect immunofluorescence studies . New data suggest that the expression of VDAC may be even more widespread . Different cell types express porin channels in their PLM and in intracellular membranes other than OMM . The functional expression of these channels may differ in the various compartments since recent experiments have demonstrated that the voltage dependence and ion selectivity of mitochondrial VDAC may be altered by their interaction with modulators . The present paper proposes a unifying concept for the ion-selective channels of cell membranes, in particular, those whose regulation is affected in cystic fibrosis.

Circ Shock, 1992 Feb, 36(2), 127 - 33
In vivo and in vitro effects of endotoxin on vascular responsiveness to norepinephrine and signal transduction in the rat; Suba EA et al.; We investigated, after in vitro and in vivo exposure to gram-negative endotoxin, the altered responsiveness of rat aortic smooth muscle to catecholamines . Two hour exposure of aortic rings from normal rats to 100 ng/ml of Escherichia coli 0111:B4 endotoxin in vitro in an artificial medium supplemented with 5% fetal calf serum at 37 degrees C did not effect the basal and norepinephrine (NE)-stimulated (10 microM, 1 hr, 37 degrees C) phosphoinositide (PI) hydrolysis and isometric contractions induced by graded doses (1 nM to 10.0 microM) of NE . Increasing the incubation time with endotoxin to 18 hr did not alter the basal PI hydrolysis but significantly (P less than 0.05) decreased the NE-induced PI hydrolysis (30% inhibition) and contractile sensitivity to NE (increase of EC50 from 20.0 +/- 3.8 to 156.4 +/- 46.7 nM) . Qualitatively similar results were obtained in experiments where rats were injected intravenously with buffer or an LD50 dose (10 mg/kg) of endotoxin . In these ex vivo measurements, only an 18 hr exposure to endotoxin caused significant (P less than 0.001) decreases in basal (58% inhibition) and NE-stimulated (75% inhibition) PI hydrolysis and in NE-induced isometric contractions (increase of EC50 from 11.0 +/- 3.3 to 664.1 +/- 280.0 nM) . The results show that the endotoxin-induced hyporeactivity to alpha 1-adrenergic receptor stimulation 1) is markedly dependent on the length of endotoxin exposure, 2) does not require (although may be enhanced by) contact with blood cells and plasma, and 3) is paralleled by a decrease in both basal and NE-stimulated PI hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Genes Dev, 1992 Feb, 6(2), 268 - 83
Bacterial cytochromes c biogenesis; Beckman DL et al.; We report the primary sequence analyses of two loci, hel and ccl, whose gene products are required specifically for the biogenesis of c-type cytochromes in the Gram-negative photosynthetic bacterium Rhodobacter capsulatus . Genetic and molecular analyses show that the hel locus contains at least four genes, helA, helB, helC, and orf52, and the ccl locus contains two genes, ccl1 and ccl2, that are essential for cytochromes c biogenesis . HelA is homologous to a class of proteins called ABC transporters and helA, helB, and helC are proposed to encode an export complex . Cytochrome c2-alkaline phosphatase gene fusions were used to show that apocytochrome c2 synthesis and secretion are not affected by the hel and ccl defects . Ccl1 and Ccl2 possess typical signal sequences to direct them to the periplasm . The periplasmic orientation of Ccl1 was confirmed using a Ccl1-alkaline phosphatase gene fusion . The Ccl1-alkaline phosphatase gene fusion analysis also demonstrated that Ccl1 does not require hel genes for its synthesis and secretion . Ccl1 is homologous to proteins encoded by chloroplast and mitochondrial genes, suggesting analogous functions in these organelles . Taken together, these results support the hypothesis that the hel-encoded proteins are required for the export of heme to the periplasm where it is subsequently ligated to the c-type apocytochromes.

Lupus, 1992 Feb, 1(2), 63 - 74
Autoimmunity, polyclonal B-cell activation and infection; Granholm NA et al.; It is widely believed that autoimmunity is an integral part of the immune system, and that genetic, immunologic, hormonal, environmental and other factors contribute to the pathogenesis of autoimmune disease . Thus, autoimmune disease may represent an abnormal expression of immune functions instead of loss of tolerance to self, and it can be organ specific or systemic in its manifestations . We review the various factors that contribute to the development of autoimmune disease; we also review the mechanisms of polyclonal B-cell activation, with emphasis on the role of infectious agents . We consider systemic lupus erythematosus in humans and in experimental animals as prototypic autoimmune disease, and we summarize data to indicate that polyclonal B-cell activation is central to the pathogenesis of systemic autoimmune disease . The effect of polyclonal B-cell activation, brought about by injections of a B-cell activator-lipopolysaccharide from Gram-negative bacteria-is sufficient to cause autoimmune disease in an immunologically normal host . In fact, autoimmune disease can be arrested if excessive polyclonal B-cell activation is suppressed; alternatively, autoimmune disease can be exacerbated if polyclonal B-cell activation is enhanced . We explore the mechanism of tissue injury when autoimmune disease is induced or exacerbated, and we consider the pathogenic roles of autoantibodies, immune complexes, complement, the blood cell carrier system, and the mononuclear phagocyte system . Although polyclonal B-cell activation may be the mechanism whereby various factors can cause or exacerbate systemic autoimmune disease, polyclonal B-cell activation may cause autoimmune disease on its own.

Biochemistry, 1992 Jan 28, 31(3), 631 - 8
Polymyxin B induces transient permeability fluctuations in asymmetric planar lipopolysaccharide/phospholipid bilayers; Schroder G et al.; The interaction of the polycationic decapeptide polymyxin B with asymmetric planar bilayers from lipopolysaccharide and phospholipid monolayers, which resemble the lipid matrix of the outer membrane of Gram-negative bacteria, was investigated . The addition of polymyxin B in micromolar amounts to the lipopolysaccharide side of the asymmetric bilayers resulted, under voltage-clamp conditions, in a fast macroscopic increase of their ionic conductance, whereas the polymyxin B nonapeptide induced no significant conductance changes . The polymyxin B induced macroscopic conductance exhibited large fluctuations and was strongly dependent on the amplitude and polarity of the transmembrane potential . The temporal pattern and amplitudes of the fluctuations were characterized by power spectra of the membrane currents and their variances, respectively . In the initial phase following peptide addition, the conductance changes appeared to be channellike discrete fluctuations . The lifetimes of the fluctuations were exponentially distributed, and the mean lifetimes were strongly voltage-dependent, ranging from approximately 30 ms at +80 mV (positive at the side opposite to peptide addition) to less than 5 ms at reverse polarity . The conductance amplitudes of the single fluctuations exhibited a broad distribution with a mean of 2 nS . A comparison of the features of the macroscopic conductance and of the discrete fluctuations showed that the former can basically be understood as a superposition of a large number of the latter . From the amplitudes of the fluctuations, the diameter of the polymyxin-induced lesions was estimated to about 3 nm . The experimental findings can be understood by assuming a detergent-like action of polymyxin B.

Br Dent J, 1992 Jan 25, 172(2), 69 - 70
DNA fingerprinting of oral bacteria; Lancashire P; This elective study was conducted at the Dental School, University of Wales College of Medicine and at the University of Wales College of Cardiff School of Pure and Applied Biology . The student's interest in oral microbiology and desire to contribute to the work being performed in the field of oral microbiology, led him to concentrate his study on developing a DNA fingerprinting method to evaluate the putative new species of the genus Capnocytophaga . The genus Capnocytophaga, which contains Gram-negative, capnophilic fusiform-shaped bacteria, has been implicated in juvenile periodontitis and prepubertal gingivitis . However, the genus is also part of the normal oral flora . The student believed that there was an implication of the existence of both pathogenic and non-pathogenic strains of Capnocytophaga and developed a method of DNA fingerprinting to distinguish Capnocytophaga strains from clusters identified by Khwaja et al . The student felt that the study allowed insight into the tremendous potential of molecular techniques for furthering the understanding of dental disease and confirmed his ambition to pursue a career in academic oral microbiology . This area of study allows new associations to be made between specific bacteria and diseases, aiding the possibility of rapid and early diagnosis.

Biochem Pharmacol, 1992 Jan 22, 43(2), 191 - 6
Endotoxin-inducible cytotoxicity in liver cell cultures--II . Demonstration of endotoxin-tolerance; Hartung T et al.; Endotoxins from gram negative bacteria, central mediators of septic shock, share the characteristic property of inducing tolerance against their own action . This work investigates whether a corresponding ex-vivo tolerance can be observed in a cellular system with endotoxin-inducible hepatocytoxicity . The following experimental approaches were chosen in order to induce an endotoxin-unresponsive state prior to cell preparation: (1) pretreatment of rats with endotoxin, (2) partial hepatectomy, (3) use of neonatal rats and (4) pretreatment of rats with silica . An in-vivo protection against endotoxin-induced liver injury was obtained by all of these four measures: cells prepared from these groups of animals showed greatly diminished sensitivity towards endotoxin-induced hepatocytotoxicity in vitro . The suppressed endotoxin sensitivity after silica pretreatment was partially restored in vitro by the addition of native Kupffer cells (KC) . Isolated KC of all but the endotoxin-pretreated animals secreted tumor necrosis factor-alpha in response to endotoxin . It is concluded that different types of tolerance can be distinguished: (a) impairment of macrophage functions (silica pretreatment), (b) hepatocyte unresponsiveness (neonatal rats and hepatectomy) and (c) impaired macrophage function combined with hepatocyte unresponsiveness (endotoxin-pretreated rats).

J Immunol, 1992 Jan 15, 148(2), 532 - 7
The role of bactericidal/permeability-increasing protein as a natural inhibitor of bacterial endotoxin; Marra MN et al.; Systemic release of endotoxin (LPS) after Gram-negative infection initiates a cascade of host cytokines that are thought to be the direct cause of shock, multisystem organ failure, and death . Endogenous LPS-binding proteins may play a role in regulating LPS toxicity in vivo . The human neutrophil granule protein bactericidal/permeability-increasing protein (BPI) shares sequence homology and immunocrossreactivity with an acute phase lipopolysaccharide binding protein (LBP) which has been shown to bind to LPS and accelerate LPS activation of neutrophils and macrophages . Although structurally similar, LBP and BPI are apparently functionally antagonistic . We previously showed that BPI inhibits LPS-mediated neutrophil activation in vitro . Here we demonstrate that BPI binds to LPS near the lipid A domain, and formation of the LPS-BPI complex abrogates detrimental host responses to LPS . For example, BPI blocks LPS-stimulated TNF release in vitro and in vivo, and LPS complexed to BPI is not pyrogenic in rabbits . Results demonstrating that BPI is released by stimulated human neutrophils further support the idea that BPI functions extracellularly in vivo to neutralize endotoxin . Taken together, these data argue that BPI neutralizes the toxic effects of LPS in vivo, and that BPI may represent a new therapeutic approach to the treatment of endotoxic shock.

Acta Microbiol Hung, 1992, 39(2), 155 - 8
The effect of kanamycin treatment of rats on the development of gastro-intestinal syndrome of radiation disease; Bertok L Jr et al.; Kanamycin pretreatment through stomach tube eliminates the endotoxin producing Gram-negative bacteria of the intestinal flora and can delay the death of rats suffering from gastro-intestinal syndrome of radiation disease . Endotoxins probably play a significant role in the pathological process of the syndrome.

Arch Surg, 1992 Jan, 127(1), 83 - 9; discussion 89
Guidelines for clinical care: anti-infective agents for intra-abdominal infection . A Surgical Infection Society policy statement; Bohnen JM et al.; Several antibiotics have been marketed for therapeutic use in intra-abdominal infection . Often, these agents do not provide a sufficient spectrum activity against both facultative and obligate anaerobic gram-negative organisms, or have certain toxic effects that would not otherwise support their use . Guidelines have been developed for selection of antibiotic therapy for intra-abdominal infections and are presented as a statement of the Surgical Infection Society endorsed by the Executive Council . These guidelines are restricted to infections derived from the gastrointestinal tract and deal with those microorganisms commonly seen in such infections . The recommendations are based on in vitro activity against enteric bacteria, experience in animal models, and documented efficacy in clinical trials . Other concerns regarding pharmacokinetics, mechanisms of action, microbial resistance, and safety were also used in the formation of these guidelines . For community-acquired infections of mild to moderate severity, single-agent therapy with cefoxitin, cefotetan, or cefmetazole or ticarcillin-clavulanic acid is recommended . For more severe infections, single-agent therapy with carbapenems (imipenem/cilastatin) or combination therapy with either a third-generation cephalosporin, a monobactam (aztreonam), or an aminoglycoside plus clindamycin or metronidazole is recommended . Regimens with little or no activity against facultative gram-negative rods or anaerobic gram-negative rods are not considered acceptable.

J Clin Microbiol, 1992 Jan, 30(1), 214 - 5
Comparison of the E test and microdilution for detection of beta-lactam-resistant mutants that are stably derepressed for type I beta-lactamase; Knapp CC et al.; The activities of cefotaxime, ceftazidime, piperacillin, and aztreonam were compared in the E test and broth microdilution test against 30 gram-negative bacterial mutants derepressed for type I beta-lactamases . The results demonstrated complete agreement between 24-h MICs of 80 to 83% and essential agreement between 24-h MICs of 90 to 97% . When sufficient growth was present for the E test to be read at 6 h, the essential agreement between 6- and 24-h E-test MICs was 100% for ceftazidime, piperacillin, and aztreonam and 85% for cefotaxime.

Am J Surg, 1992 Jan, 163(1), 174 - 9; discussion 179-80
Cytokine regulation of intestinal glutamine utilization; Austgen TR et al.; The effects of cytokines on intestinal glutamine metabolism were studied to gain further insight into the regulation of altered glutamine metabolism that occurs during severe infection . One hundred thirteen adult rats were given a single dose of interleukin-1 (IL-1, 50 micrograms/kg), tumor necrosis factor (TNF, 50 micrograms/kg or 150 micrograms/kg), or saline (controls), and flux studies were performed 4 or 12 hours later . Intestinal blood flow was not different between control and cytokine-treated animals at either time point . At the 4-hour time point, arterial glutamine fell by 16% to 21% in the cytokine-treated animals (p less than 0.05); at the 12-hour time point, the arterial glutamine concentration had returned to normal . Intestinal glutamine extraction decreased in the animals treated with IL-1 at both time points (4 hours: 13% +/- 1.3% in IL-1 versus 20% +/- 1.6% in controls, p less than 0.05; and 12 hours: 9% +/- 2% in IL-1 versus 17% +/- 2% in controls, p less than 0.05) . Consequently, net intestinal glutamine uptake fell in the animals treated with IL-1 at both time points (p less than 0.05) . Similarly, the activity of mucosal glutaminase, the principal enzyme of glutamine hydrolysis in the gut, fell by 50% in the 4-hour study (6.1 +/- 0.6 mumol/h/mg protein in IL-1 versus 9.6 +/- 0.8 mumol/h/mg protein in controls, p less than 0.01) and by 40% in the 12-hour study (5.4 +/- 0.5 mumol/h/mg protein in IL-1 versus 8.8 +/- 0.4 mumol/h/mg protein in controls, p less than 0.05) . Concomitant with the aforementioned decrease in gut glutamine metabolism was a 25% incidence of positive blood cultures for gram-negative organisms in IL-1 treated rats studied at the 12-hour time point (p = 0.05 versus controls) . In the doses administered and at the time points studied, TNF had no effects on the parameters of gut glutamine metabolism examined . The results indicate that IL-1 is a potential mediator of the alterations in gut glutamine metabolism observed in sepsis and endotoxemia.

Genetics, 1992 Jan, 130(1), 27 - 36
Promoters of the broad host range plasmid RK2: analysis of transcription (initiation) in five species of gram-negative bacteria; Greener A et al.; A broad host range cloning vector was constructed, suitable for monitoring promoter activity in diverse Gram-negative bacteria . This vector, derived from plasmid RSF1010, utilized the firefly luciferase gene as the reporter, since the assay for its bioluminescent product is sensitive, and measurements can be made without background from the host . Twelve DNA fragments with promoter activity were obtained from broad host range plasmid RK2 and inserted into the RSF1010 derived vector . The relative luciferase activities were determined for these fragments in five species of Gram-negative bacteria . In addition, four promoters were analyzed by primer extension to locate transcriptional start sites in each host . The results show that several of the promoters vary substantially in relative strengths or utilize different transcriptional start sites in different bacteria . Other promoters exhibited similar activities and identical start sites in the five hosts examined.

Crit Rev Oral Biol Med, 1992, 3(1-2), 31 - 60
Host mediators in gingival crevicular fluid: implications for the pathogenesis of periodontal disease; Lamster IB et al.; During the past few years, a considerable number of studies have examined different aspects of the host response in gingival crevicular fluid (GCF), including the relationship of specific markers to the active phases of periodontal disease . Various indicators of the acute inflammatory response (the lysosomal enzymes beta-glucuronidase and collagenase, the cytoplasmic enzyme aspartate aminotransferase, and the arachidonic acid metabolite PGE2) have been shown to be associated with clinical attachment loss in chronic adult periodontitis in man and experimental periodontitis in animal models . In contrast, the relationship of indicators of the humoral immune response in GCF to active periodontal disease is equivocal . Furthermore, a number of indicators of the cellular immune response have been identified recently in GCF (i.e., Interleukin-1 alpha, IL-1 beta, tumor necrosis factor-alpha), but their relationship to active phases of periodontal disease have not been studied . The polymorphonuclear leukocyte (PMN) is the cellular hallmark of acute inflammation . Evidence from the GCF studies suggests that hyperreactivity of these cells plays a critical role in the active phases of some forms of periodontal disease . Metabolic activation of PMN can be associated with a number of potentially destructive reactions . The major effector mechanism for tissue destruction that can be specifically identified with the PMN is the synergistic effect of the release of PMN proteases and the generation of reactive oxygen metabolites by these cells . Priming of the PMN, where the PMN response is enhanced by agents that do not initiate the response, may be an important mechanism for PMN activation in the crevicular environment; for example, cytokines such as IL-1 beta and TNF-alpha, and lipopolysaccharides released from subgingival Gram-negative bacteria, can serve this function . The hypothesis proposed here argues that in addition to the severe forms of periodontal disease that have been associated with qualitative or quantitative PMN defects, tissue destruction in the periodontum can be observed with hyperreactivity of these cells . These differing conclusions do not create a dilemma, but may represent opposite ends of a balance that is no longer in equilibrium.

J Immunol, 1992 Jan 1, 148(1), 234 - 8
A nontoxic, idiotope vaccine against gram-negative bacterial infections; Su S et al.; Experiments were performed to test the ability of mouse antiidiotopic mAb, specific for an antilipid A mAb, to act as a vaccine against gram-negative bacterial infections . Lipid A is a conserved region of bacterial LPS . Immunization with the antiidiotopic antibodies, coupled to an immunogenic carrier protein (hemocyanin), specifically induced anti-LPS antibody responses in animals from different species . In a mouse model, this immunization resulted in protection against both lethal gram-negative bacteremia and endotoxemia . The antiidiotopic antibodies, however, did not stimulate endotoxin-associated bioactivities, such as induction of TNF and IL-1 . These results support the hypothesis that an idiotope vaccine can stimulate beneficial protective immunity against gram-negative infections without the toxicity inherent in LPS.

Exp Clin Immunogenet, 1992, 9(1), 48 - 56
Complement-mediated killing of susceptible gram-negative bacteria: an elusive mechanism; Taylor PW; Exposure of gram-negative bacteria to a source of antibody and complement frequently results in efficient cell killing; this effect follows the deposition of C5b-9 membrane attack complexes onto the bacterial surface . This review examines the molecular and physiological events culminating in C5b-9 deposition and cell death and advances an hypothesis that may account for cytoplasmic-membrane damage following complement-mediated perturbation of the outer membrane.

Res Exp Med (Berl), 1992, 192(3), 197 - 204
Reduced clearance of leukocytes from lungs in septic rats; Hansson L et al.; Leukocytes have previously been shown to sequestrate in the lungs and liver in association with traumatic and septic shock . In a rat model of gram-negative sepsis of intra-abdominal origin, a previously described in vivo technique was used for dynamic studies of leukocyte sequestration in different organs using white blood cells labeled with 111-indium-oxine . One group of rats was either studied immediately after induction of sepsis or for 6 h under a scintillation camera for continuous registration of the activity distribution (i.e., presence of leukocytes) . Another group was studied 12 h after induction of sepsis for 60 min . The activity increased immediately over the lungs, indicating sequestration of the leukocytes during the first 6 h, but there was no significant difference in this respect between septic and control animals . It does not seem possible to study leukocyte sequestration dynamically in this way . When the labeled leukocytes were administered 12 h after induction of sepsis, however, the activity of septic animals' lungs was seen to remain elevated over the time period studied compared with control rats, in which the activity slowly decreased . In the liver and spleen, the activity increased in both groups, but significantly more so in control animals, which may be explained by disturbed leukocyte margination and cell turnover in the septic animals . This study has indicated that leukocyte distribution in different organs is affected by sepsis and this reaction can be studied using radiolabeled leukocytes.

Patol Fiziol Eksp Ter, 1992 Jan-Feb, (1), 23 - 6
{Protective effect of unithiol and its combination with magnesium sulfate in endotoxin poisoning and in infection by gram-negative bacteria}; Grutman MI et al.; It is shown that unithiol protects mice from death in intoxication by S . typhimurium and S . Sonnei lysates and in infection caused by S . typhimurium . Magnesium sulfate strengthens the protective effect of unithiol . It is suggested that the protective effect of unithiol is due to its antioxidant action.

Dig Dis, 1992, 10(3), 144 - 52
'Gastrospirillum hominis', another gastric spiral bacterium; Fischer R; 'Gastrospirillum hominis' is a 'new' tightly coiled gram-negative bacterium carrying bundles of sheathed polar flagella . It has been rather infrequently detected in antral and, even more rarely, in fundic mucosa samples removed at endoscopy from patients investigated for Helicobacter pylori colonization . Until now, it has remained noncultivable but has successfully been maintained in laboratory mice . Its identity with similar bacteria found in the stomachs of cats, dogs, monkeys, pigs, and other animals is uncertain . It was probably already seen by early investigators in the first half of this century . Preliminary data published in case reports suggest that it is associated with more or less active chronic antral gastritis, that it is restricted to the gastric epithelium, and that it possesses a urease, thus limiting the specificity of urease tests for H . pylori . There is hitherto no solid proof that it can induce inflammation although it seems capable of invading parietal and other glandular cells and causing ultrastructural changes . Similar organisms spontaneously colonizing the stomachs of rhesus monkeys were shown to increase gastric acid secretion.

Arkh Patol, 1992, 54(1), 9 - 14
{Pathology and pathogenesis of sepsis}; Permiakov NK; The key problems of the theory of sepsis and criteria of its postmortem diagnosis are discussed on the basis of pathological and bacteriological study of about 4000 purulent-septic diseases observed for the last 40 years in the pathology department of N . V . Sklifosovsky Moscow Research Institute of Emergency Medical Aid (sepsis after abortion, surgical and iatrogenic sepsis, acute septic endocarditis, purulent peritonitis, mediastinitis, pleuritis, phlegmons of body and limbs, bacterial shock, etc.) . Sepsis, according to the author, is a generalized infectious disease developing acyclically, produced mainly by purulent coccal flora and having the course of septicopyemia . A metastatic purulent focus is an obligatory sign of the generalization . Septicemia is a local inflammatory process produced mainly by bacterial gram-negative flora . It can be a prologue of sepsis but more frequently develops in two directions: 1) purulent-resorptive fever with an acute, subacute and chronic course; 2) bacterial shock with a fulminant course and high lethality.

Urol Int, 1992, 48(2), 191 - 4
Diffusion of cefmenoxime and latamoxef into prostatic fluid in the patients with acute bacterial prostatitis; Katoh N et al.; Twenty-two patients with acute bacterial prostatitis were treated with cefmenoxime (CMX) or latamoxef (LMOX), which have susceptibilities against various gram-negative bacteria . First 11 patients received a 5- to 12-day course of cefmenoxime and the next 11 received a 6- to 13-day course of latamoxef . All patients were treated successfully except 1 patient with a drug allergy . Diffusion of CMX or LMOX into prostatic fluid in these patients and healthy controls were evaluated . The mean value of CMX in the expressed prostatic fluid was 12.8 micrograms/ml in the patients receiving 2 g of CMX intravenously and 0.7 micrograms/ml in the controls . The mean value of LMOX was 14.0 micrograms/ml in the patients receiving 2 g of LMOX intravenously and 1.2 micrograms/ml in the controls . The diffusion of CMX and LMOX into prostatic fluid in the patients with acute bacterial prostatitis was strikingly higher than that of controls.

Eur Cytokine Netw, 1992 Jan-Feb, 3(1), 7 - 17
Anti-cytokine strategies; Dinarello CA; Although antibiotics and support systems have reduced the mortality due to bacterial infections, 40-50% mortality is still an unacceptable statistic . Anti-LPS (anti-lipid A) passive immunotherapy has reduced this mortality but only in patients with documented Gram-negative bacteriemia . However, anti-cytokine therapy such as monoclonal antibodies to TNF and the IL-1ra have reduced mortality to all infectious causes . IL-1ra is presently in Phase III trials as is monoclonal anti-TNF . Two other strategies are soluble (extracellular domains) receptors to TNF and IL-1 . These are now entering Phase I trials but animal data strongly support that similar to antibodies to TNF and IL-1ra, these anti-cytokine therapies will also be effective.

Electron Microsc Rev, 1992, 5(2), 381 - 419
Uptake, distribution and fate of bacterial lipopolysaccharides in monocytes and macrophages: an ultrastructural and functional correlation; Kang YH et al.; Bacterial lipopolysaccharides (LPS), which are important components of the cell wall of gram-negative bacteria, induce a number of host responses both beneficial and harmful . The present review elucidates the uptake, distribution and functions of LPS in mononuclear phagocytes in an attempt to gain an insight into the mechanisms which control the pathogenesis of LPS mediated septic shock . The unique feature of LPS bilayer structure, the tagged LPS and antibodies to LPS provide means for studying binding, uptake, fate and subcellular distribution of LPS in tissues and cells . LPS bind to monocytes and macrophages by specific interaction via receptors such as scavenger receptors, CD14 and CD18 and by non-specific interactions, and enter the cells via receptor-mediated endocytosis, absorptive pinocytosis, phagocytosis, and diffusion . The ingested LPS are localized in pinocytic vesicles, phagocytic vacuoles, cytoplasm, mitochondria, rough endoplasmic reticulum, Golgi apparatus, and nucleus . The interactions of LPS with monocytes and macrophages trigger a broad spectrum of cellular responses, including production of important bioactive factors or mediators, such as IL-1, TNF, interferons, prostaglandins, and macrophage-derived growth factor, which are implicated in the pathogenesis of septic shock and wound healing . However, there is no conclusive evidence indicating that production of the mediators can only be induced through specific interactions.

J Dairy Sci, 1992 Jan, 75(1), 78 - 84
Field trial to determine efficacy of an Escherichia coli J5 mastitis vaccine; Hogan JS et al.; Efficacy of an Escherichia coli (O111:B4) J5 bacterin for preventing naturally occurring IMI and clinical mastitis was tested in a 2.5-yr field trial in a 225-cow commercial herd . Cows with odd-numbered identification were vaccinated, and cows with even-numbered identification served as unvaccinated controls for each lactation during the study . Immunizations were subcutaneous on the upper part of the rib cage just posterior to the scapula at drying off, 30 d after drying off, and at calving . Percentage of quarters infected at calving with Gram-negative bacteria did not differ between treatment groups . A total of 67% of Gram-negative bacterial IMI present at calving in control cows became clinical during the first 90 d of lactation compared with 20% in vaccinated cows . Rate of Gram-negative bacterial clinical mastitis was higher in control cows than in vaccinated cows during the first 90 d of lactation . Immunization with the E . coli J5 bacterin did not reduce level of Gram-negative bacterial IMI at calving but did reduce incidence of clinical mastitis.

Biophys J, 1992 Jan, 61(1), 172 - 88
Two-dimensional crystallization of a bacterial surface protein on lipid vesicles under controlled conditions; Paul A et al.; The solubilized surface protein of the Gram-negative bacterium Comamonas acidovorans was reconstituted on lipid vesicles by means of controlled dialysis . To this end, a multichamber dialysis apparatus was built which allows one to control the temperature and the dialysis rate, to apply various temperatures or buffer systems and sample conditions in a single experiment, and to monitor the turbidity of the sample by means of light scattering . The reconstitution conditions were optimized such that the surface protein formed two-dimensional crystals suitable for electron crystallography . The recrystallized surface protein arrays gave a resolution of approximately 1.3 nm in projection after correlation averaging of negatively stained preparations . The surface protein assembled into partially self-contained two-dimensional crystals which possess a strong shape-determining effect and formed cylinders and various cone-shaped vesicles . The development of the various vesicle forms is described in a model.

Intensive Care Med, 1992, 18(5), 293 - 8
Recurrent episodes of gram-negative bacteremia or endotoxemia change reactivity of pre- and post-capillary pulmonary segments to angiotensin or free radicals; Kirton OC et al.; OBJECTIVE: Recurrent episodes of Gram-negative bacteremia (from intraperitoneal abscesses) or endotoxemia cause lung microvascular injury in the rat . Change in vascular reactivity was assessed in response to challenge . DESIGN: In the isolated lung preparation, resistance was partitioned between pre-(PVRa) and post-capillary (PRVv) segments: vasoreactivity was assessed by challenge with Angiotensin II (AII) or reactive oxygen metabolites . Animals received 4 weekly intra-abdominal implants of live E . coli and B . fragilis in a carrier of sterile cecal content and barium sulfate (SEPSIS) or carrier alone (SHAM SEPSIS): or 4 weekly intravenous infusion of E . coli endotoxin (ENDO) or of saline (SHAM ENDO) . A fifth group were untreated controls (CONTROL) . MEASUREMENTS AND RESULTS: In the SEPSIS and ENDO lungs, PVRa and PVRv before challenge were normal . In the SEPSIS lung, AII increased PVRa more than in the control lung, PVRv to a similar degree in both . In the ENDO lung it increased PVRa compared with its effect on the SHAM ENDO lung: In both it also increased PVRv, to a similar degree and well above the baseline . Always tachyphylaxis developed with increases in dosage (to 25 microns and 50 microns, respectively) . Oxygen free radical challenge in the SEPSIS and ENDO lung caused significant vasoconstriction, particularly PVRv, whereas no response was observed in the CONTROL or SHAM-treated lung from either group . CONCLUSION: Abnormal lung vascular reactivity after SEPSIS or ENDOTOXIN is evident on challenge, the two agents used here detecting site specific changes.

Acta Vet Scand, 1992, 33(2), 129 - 38
The role of endotoxins in methscopolamine induced ruminal stasis in calves; Aiumlamai S et al.; Methscopolamine was used to induce ruminal stasis in calves . Clinical and blood biochemical parameters were studied to judge the possible role of gastro-intestinal endotoxins from Gram-negative bacteria . Two trials were carried out where one injection of 100 mg and 3 consecutive injections of 70 mg of methscopolamine were administered . The animals showed signs of ruminal stasis . General clinical signs and changes in blood biochemical parameters were similar to what is found in endotoxaemia or in induced ruminal acidosis . Relevant parameters such as prostaglandin F2 alpha metabolite, endotoxin, iron, zinc, calcium and glutamate dehydrogenase changed significantly indicating exposure of endotoxins.

Acta Vet Scand, 1992, 33(2), 117 - 27
The role of endotoxins in induced ruminal acidosis in calves; Aiumlamai S et al.; Experimentally induced ruminal acidosis was carried out in 4 calves to investigate the possibility of resorption of endotoxins from Gram negative bacteria originating from the gastro-intestinal tract . The ruminal acidosis was induced by overfeeding of oats and the effect was evaluated by clinical and blood biochemical changes . Blood samples were collected every 2 h for 60 h before and after experimental feeding . The animals showed signs of ruminal acidosis and also the clinical and blood biochemical changes were similar to those seen after experimental endotoxaemia . However, although being less dramatic the changes in many relevant parameters such as an increase in prostaglandin F metabolite levels, body temperature, endotoxins and a decrease in iron indicate that an endotoxaemic state had occurred . The results of the present study show that ruminal acidosis/stasis is linked to resorption of endotoxins from the gastro-intestinal tract causing endotoxaemia/endotoxicosis.

Annu Rev Microbiol, 1992, 46, 141 - 63
Genetics of ribosomally synthesized peptide antibiotics; Kolter R et al.; In recent years many peptide antibiotics have been shown to be ribosomally synthesized . Among these are many microcins, produced by diverse strains of gram-negative bacteria . While the structures and modes of action of these peptide antibiotics vary widely, many of them share several important features . Their synthesis is often induced by the cessation of growth . In addition, many of them undergo unusual posttranslational modifications to yield the mature molecule, which is often exported from the cell by a dedicated export apparatus . The genes involved in modification and export of the peptide antibiotics are generally found adjacent to the structural gene and are under the same regulation . The results supporting these conclusions are reviewed and discussed in this chapter.

Przegl Lek, 1992, 49(5), 151 - 3
{Sensitivity of multiresistant bacterial strains to imipenem (Tienam)}; Ruczkowska J et al.; The susceptibility to imipenem of 213 clinical strains of Gram-negative rods resistant to six cephalosporins (cephradine, cefamandole, cefuroxime, cefotaxime, cefoperazone and ceftriaxone) has been studied . It was found that 93.9% of the strains were sensitive and 6.1% were resistant to imipenem . In the examined group 64 (30%) out of 213 strains were simultaneously resistant to four aminoglycosides (gentamycin, tobramycin, netilmicin and amikacin); among them 5 (7.8%) strains were resistant to imipenem . Sensitivity to imipenem 157 out of 213 examined strains as compared to that of ceftazidime was as follows: 29.3% of the strains were sensitive and 3.8% resistant to both of the antibiotics, whilst 64.3% were imipenem-sensitive and ceftazidime-resistant . In vitro imipenem proved to be very active antibiotic against over 90% of multiresistant Gram-negative rods.

Circ Shock, 1992 Jan, 36(1), 45 - 56
Loss of microvascular negative charges accompanied by interstitial edema in septic rats' heart; Gotloib L et al.; We studied the effect of Gram-negative sepsis on negative charges of heart capillaries and myocardial cells . We used a rat model of multiorgan failure, with ruthenium red (RR) and polyethyleneimine (PEI) as cationic binding tracers . Twenty-four hours after induction of sepsis, negative charges had decreased in glycocalyx and basement membrane of myocardial capillary endothelial cells . There were substantial amounts of interstitial edema . Density of anionic charges in the sarcolemmal glycocalyx complex of cardiac cells was markedly reduced . Myocardial cells' mitochondria consistently showed morphologic changes, whose severity ranged between stages II and IV C of Trump . Thirteen days after induction of sepsis, capillary endothelial and myocardial cells had recovered almost completely and showed no intracellular edema . Gram-negative sepsis caused a significant reduction in negative charges normally present in the microvascular wall as well as on myocardial cells . Consequently, several membranes limiting the various compartments of heart tissue lost their structural integrity . This morphometric data could explain the development of protein-rich interstitial edema and defective cell volume regulation observed in cardiac muscle of endotoxin-shocked animals . This myocardial edema may be at the origin of the cardiac dysfunction observed in both experimental and human septic shock.

Infect Immun, 1992 Jan, 60(1), 231 - 6
Roles of protein kinase C and G proteins in activation of murine resting B lymphocytes by endotoxin-associated protein; Bandekar JR et al.; Endotoxin-associated protein (EP) from the outer membrane of gram-negative bacteria is a potent immunomodulator . To examine the mechanism of EP stimulation, the protein kinase C inhibitors H7 and staurosporine were used . Both DNA and RNA synthesis of EP-stimulated murine resting B cells were completely inhibited when inhibitors were added at 0 h, whereas 55 to 76% inhibition of DNA synthesis was observed when H7 was added after 12 h of stimulation . In contrast, HA 1004, which blocks protein kinase A and protein kinase G activity, was relatively ineffective even at high concentrations, suggesting that the activity of protein kinase C is a primary mechanism of EP-induced murine B-cell proliferation . To examine the role of G proteins in EP-induced DNA synthesis in B cells, the effects of pertussis toxin (PT), which inactivates certain G proteins, and the B oligomer of PT (PTB), which does not, were also examined . PT was found to inhibit EP-induced DNA synthesis in a dose-dependent manner . However, PTB also caused equivalent inhibition, suggesting that PTB may be responsible for most of the inhibitory effect seen with the holotoxin . These results serve to question whether G proteins are involved in the signal transduction that occurs during EP-induced DNA synthesis in murine B cells.

Adv Perit Dial, 1992, 8, 39 - 41
Increased uptake of radio labelled white blood cells into abdomen of uremics on peritoneal dialysis; Malacarne F et al.; Peritonitis and exit-site tunnel infection are frequent causes of CAPD drop out . We studied 9 patients, 8 treated by CAPD and 1 by IPD . These patients underwent sonographic and scintigraphic study of the abdomen . All scintigraphic examinations showed a visceral uptake . In two cases, sub-clinical bowel inflammation, demonstrated by scintigraphic study, preceded a gram negative peritonitis . The scintigraphic study with radiolabelled white blood cells may be useful in identifying chronic aseptic inflammations and some bowel and exit-site conditions which are possible risk factors in some cases of peritonitis.

Adv Perit Dial, 1992, 8, 230 - 3
Erythema: does it indicate infection in a peritoneal catheter exit site?
Gonthier D, Bernardini J, Holley JL, Piraino B.
The definition of a peritoneal catheter exit site infection varies from one dialysis center to another . A review of abnormal appearing exit sites (n = 334 in 169 patients) from 1/83 to 3/91 was done to compare outcome in exit sites presenting with erythema (39, 12%) to those with drainage plus erythema (72, 22%) or drainage alone (223, 67%) . Resolution of the abnormality occurred in 48% of those exit sites with drainage, 62% with erythema and drainage, and 79% with erythema alone (p < 0.005) . S . aureus was present in 62% of the cultured exit sites which had erythema alone, 64% with erythema plus drainage, and 41% with drainage alone, while Gram negative rods were present in 13%, 12%, and 35%, respectively (p < 0.005) . Twenty-three of the 39 exit sites with erythema were not initially treated with antibiotics; 87% resolved compared with 69% of those treated immediately with antibiotics (p = ns) . Seven of the 8 erythematous exit sites that did not resolve progressed to tunnel infection and/or peritonitis and required catheter removal, despite the addition of antibiotics in the three initially untreated . Six of the 8 unresolved erythematous exit sites were due to S . aureus . These results indicate that, although drainage is the commonest exit site abnormality and has the worst prognosis, peri-catheter erythema is not always benign, representing an early sign of infection in some cases.

Rev Pneumol Clin, 1992, 48(2), 75 - 8
{Acute respiratory insufficiency caused by hyperinfestation with strongyloides . BALF diagnosis and favourable outcome}; Mejia JH et al.; Hyperinfestation with Strongyloides is a severe complication in immunodepressed patients . It may present with various clinical signs, notably acute respiratory failure . Diagnosis may be difficult, particularly when the strongyloidiasis is associated with septicaemia caused by Gram-negative organisms . We report a new case of hyperinfestation with Strongyloides in a patient treated for periarteritis nodosa . This case was remarkable on two scores: the diagnosis problem raised by the presence of intrapulmonary haemorrhages, and the favourable outcome of an acute and initially severe respiratory failure which had required assisted ventilation . The role played in the patient's cure by the doses of thiabendazole given and the duration of their administration is discussed.

Histopathology, 1992 Jan, 20(1), 67 - 9
Renal botryomycosis; Richmond I et al.; A case of visceral botryomycosis which arose in the left kidney of a 60-year-old woman is reported . This is the fifth reported case of renal botryomycosis which, in this patient, was mistaken clinically for a renal carcinoma . The lesion was composed of confluent abscesses containing 'sulphur granule-like' lesions in which irregularly lobed aggregates of Gram-negative organisms surrounded by an eosinophilic capsule were noted . Awareness of this unusual lesion is important because of its histopathological similarity to actinomycosis.

Virology, 1992 Jan, 186(1), 175 - 91
Deletion of the growth factor gene related to EGF and TGF alpha reduces virulence of malignant rabbit fibroma virus; Opgenorth A et al.; The role of the epidermal growth factor homologue in malignant rabbit fibroma virus (MRV) pathogenicity was investigated by constructing a viral growth factor deletion mutant (MRV-GF-) . Since MRV is a recombinant virus with a myxoma virus background but possesses some terminal sequences derived from Shope fibroma virus, the growth factor gene in MRV is in fact identical to Shope fibroma growth factor (SFGF) . Although no significant differences were detected in the in vitro characteristics of MRV and MRV-GF-, a pronounced attenuation was observed after inoculation of the test rabbits with MRV-GF- . Animals infected with wild-type MRV uniformly developed a fatal syndrome involving disseminated tumors accompanied by purulent conjunctivitis and rhinitis . In contrast, although MRV-GF- recipients developed similar initial signs of the MRV disease syndrome, 75% of these animals completely recovered from the viral and secondary bacterial infections and became immune to subsequent MRV challenge . Tumors in MRV-GF- recipients displayed earlier and more prominent inflammatory reactions than their wild-type MRV counterparts and contained fewer proliferating cells . Squamous metaplasia and hyperplasia of target epithelia were less pronounced in MRV-GF- than in MRV infection . We conclude that SFGF is a major virulence factor in MRV infection and is responsible for at least some of the cellular proliferation observed at tumor sites . In addition, the diminished ability of MRV-GF- to cause hyperplasia in nasal and conjunctival epithelia may decrease the extent of gram negative bacterial overgrowth as compared to the parental virus and hence contribute to the dramatic reduction in the lethality of MRV-GF- infection.

Acta Vet Hung, 1992, 40(3), 195 - 201
Early damages to lung capillaries in enzootic pneumonia of rabbits; Iregui C et al.; The present study is part of a series of investigations aimed at characterizing the "enzootic pneumonia" phenomenon in rabbits, both from the clinical-epidemiological and the pathophysiological point of view . All affected rabbits included in this study showed an acute injury to pulmonary microvasculature, similar to changes reported in pulmonary infections caused by Gram-negative bacteria . Similar alterations were not found in the control rabbits . The clinical and morphological findings revealed certain similarities between the Enzootic Pneumonia Syndrome of rabbits and Adult Respiratory Distress Syndrome (ARDS) of man . Therefore, it is suggested that, just as in the human syndrome, heterophils (PMN) are directly involved in the pathogenesis of the process . Many of the similarities noted between the two syndromes would make it possible to postulate the natural disease in rabbits as an eventual animal model suitable for extrapolating data to humans.

Gig Tr Prof Zabol, 1992, (7), 15 - 7
{Immunological studies in byssinosis from exposure to a mixed plant dust (jute, flax and kenaf}; Zaritskaia LP; Complex immunologic studies using 1 and 2 level tests in 221 individuals (171 workers of raw material processing shops and 50 volunteers) revealed the similar changes in immunologic status of people suffering from byssinosis and workers of long length of service: decreased T-lymphocyte level (owing to T-helpers), phagocyte count and increased serum IgA, IgM in comparison with healthy people . Depressed phagocytosis in 79.6% of people facing byssinosis and 71% of workers with the long length of service can be caused by Gram negative bacteria, contaminating the air of industrial premises . Workers with the short length of service showed increased T-cells level (owing to T-suppressors) and lowered serum IgA, which can be considered as a primary immune response to the dust.

Ann Trop Paediatr, 1992, 12(4), 433 - 40
Bacteraemia in severely malnourished children; Friedland IR; During a 2-year period, 792 severely malnourished children (kwashiorkor, marasmus and marasmic kwashiorkor) were hospitalized (14.8% of all admissions) . Community-acquired bacteraemia was recorded in 7.7% and nosocomial bacteraemia in 2.2% of malnourished children . The relative risk for bacteraemia on admission was 1.6 (95% confidence interval {CI} 1.3-1.9) and 2.0 for nosocomial bacteraemia (95% CI 1.4-3.0) in malnourished children as compared with well nourished children . Bacteraemia was 3.5 times more common in children with kwashiorkor than in marasmic children . Gram-negative enteric aerobes were isolated 2.6 times more frequently from malnourished than from well nourished children . The relative risk of death in malnourished children with bacteraemia was 2.5 times that of malnourished children without bacteraemia . Thirty-six per cent of deaths in children with kwashiorkor were related to bacteraemia . This study emphasizes the increased risk of bacteraemia in malnourished children, particularly those with kwashiorkor, and the impact on mortality.

Ned Tijdschr Geneeskd, 1991 Dec 28, 135(52), 2482 - 5
{Epidemiological surveillance of antibiotic resistance in a general hospital for the assessment of 'blind' antibiotics policy}; van Woensel JB et al.; A study was made of the epidemiology of gentamicin-resistant Gram-negative rods in a large general hospital over the last 12 years, and of the consequences of the presence of these bacteria for antibiotics policies, especially the use of gentamicin . Resistant bacteria were endemic at a low level: of the clinical isolates, 670 from 488 patients contained resistant bacteria, 2% of all Gram-negative rod isolates . Sporadically there were minor epidemics . Especially seriously ill, elderly patients became infected . Every year multiresistant bacteria contributed to a number of deaths . The choice of gentamicin as a drug of first, blind, choice in life-threatening infections remained justified . Continuous monitoring of Gram-negative rods is an important instrument for a preventive policy.

JAMA, 1991 Dec 25, 266(24), 3466 - 71
Cost-effectiveness of HA-1A monoclonal antibody for gram-negative sepsis . Economic assessment of a new therapeutic agent; Schulman KA et al.; OBJECTIVE--To assess the cost-effectiveness of the HA-1A monoclonal antibody for the treatment of gram-negative bacteremia . DESIGN--Cost-effectiveness analysis of a randomized, double-blind, placebo-controlled trial using clinical efficacy data reported in the medical literature . SETTING--Hospitalized patients with sepsis . PATIENTS--543 patients with sepsis and suspected gram-negative infection . Patients enrolled in the study met strict criteria for sepsis, including fever or hypothermia (less than 35.6 degrees C or greater than 38.3 degrees C), tachycardia (greater than 90 beats per minute), tachypnea (greater than 20 breaths per minute), and hypotension or two of six signs of systemic toxicity . INTERVENTION--HA-1A vs placebo in addition to usual care . MAIN OUTCOME MEASURES--We determined the range of possible cost-effectiveness ratios for HA-1A therapy, using modeling techniques when clinical or economic variables were unknown . We subjected the model to rigorous sensitivity analysis . We calculated the incremental cost of care and years of life saved for patients with sepsis and modeled two different treatment strategies: treat all patients with sepsis or test and then treat only patients with positive test results . $24,100 per year of life saved based on the treat strategy and $14,900 based on the test strategy . In sensitivity analysis the ratios ranged from $5200 to $110,200 per year of life gained . Annual costs of care for these two strategies are $1.3 billion for the test strategy and $2.3 billion for the treat strategy . CONCLUSION--Economic assessment of new technologies early in their development can be used to guide their efficient clinical introduction.

J Biol Chem, 1991 Dec 25, 266(36), 24446 - 50
Plasticity of Escherichia coli porin channels . Dependence of their conductance on strain and lipid environment; Buehler LK et al.; The conductance properties of three members of the porin family which form channels across the outer membrane of Gram-negative bacteria were compared . With their endogenous lipopolysaccharide (LPS) bound, the closely related porins F and C from Escherichia coli reveal significantly different conductance steps and closing potentials, with values of 0.82 nS (nanosiemens) and 89 mV for F-type channels, and 0.49 nS and 158 mV for C-type pores (1 M NaCl), respectively . On the basis of their closing potentials, the two channel types can be distinguished unequivocally . If reconstituted in asolectin and extraneous LPS, porin C forms F-type in addition to C-type channels . Substitution of asolectin by mitochondrial lipids yields the native C-type pores only . Both channel types can be induced to assume the mutually other channel configuration by variation of ionic strength . A multiplicity of channel subtypes is observed by variation of the pH of the medium . The three channels within a trimer are, however, consistently of the same type . Since structural studies have revealed a single channel per monomer, the several conductance steps observed are likely to reflect distinct configurations of the same channel . Best channel recoveries were observed if endogenous LPS remained associated to porin during purification . Significant yields could nevertheless be obtained also if LPS was removed from porin and replaced with various precursors or chemically synthesized analogues . As function requires the presence of glycolipids, yet crystallization is perturbed by heterodisperse endogenous LPS, the smallest monodisperse analogues yielding good channel recovery were determined . The minimal synthetic moiety is a monoglucosaminetetraacyl compound . The characteristics of porin B from E . coli BE are shown to be indistinguishable from those of porin F . The conductance properties of this porin, refolded from random coil configuration, are indistinguishable from those exhibited by native protein . The formation of channels is thus encoded by the sequence of the mature polypeptide alone.

Gene, 1991 Dec 20, 109(1), 167 - 8
An improved Tn7-based system for the single-copy insertion of cloned genes into chromosomes of gram-negative bacteria; Bao Y et al.; A system is described for the single-copy, stable insertion of cloned DNA sequences into the chromosomes of Gram- bacteria . Two narrow-host-range plasmids form the basis of this system: the 'carrier' plasmid contains the mini Tn7-Km transposon, into which foreign DNA can be cloned; the 'helper' plasmid provides the Tn7 transposition functions in trans . Both plasmids are readily transferred into Gram- bacteria by conjugation . The functionality of this system has been demonstrated in Rhodospirillum rubrum.

FEBS Lett, 1991 Dec 2, 294(1-2), 43 - 6
In vivo study of the state of order of the membranes of gram-negative bacteria by Fourier-transform infrared spectroscopy (FT-IR); Schultz C et al.; Temperature-induced order/disorder transition profiles were obtained from the membranes of intact Gram-negative bacterial cells by FT-IR analysis of the frequency shifts of the acyl chain methylene symmetric stretching band as a monitor . Cells grown at different temperatures yielded distinct transition profiles . At the individual growth temperatures, however, the nearly alike frequency values indicated a very similar 'state of order' of the bacterial membranes . The FT-IR data were complemented by GC analysis of whole cell fatty acid composition . The FT-IR data obtained in vivo gave direct evidence of the adaptation of the 'state of order' and 'fluidity' of bacterial membranes to varying growth temperatures.

Crit Care Med, 1991 Dec, 19(12), 1480 - 5
Effect of individualized pharmacokinetic dosing on patient outcome; Whipple JK et al.; OBJECTIVE: To study the effect of individualized pharmacokinetic dosing of aminoglycosides on patient outcome . DESIGN: Prospective, randomized study . SETTING: Tertiary care hospital . PATIENTS: Ninety-five patients with documented Gram-negative infections received 97 courses of aminoglycoside therapy . INTERVENTIONS: Patients were randomized between pharmacokinetic dose adjustment and monitoring or traditional physician-directed techniques . Patients were stratified by severity of underlying illness before randomization . MEASUREMENT AND MAIN RESULTS: Sixty-two courses of treatment were satisfactorily completed . Patients in the severely ill group (eight kinetic, eight traditional) had significantly (p less than .05) better survival (7 kinetic, 3 traditional) when managed with pharmacokinetic consultation . The kinetic arm received greater doses (156 +/- 59 mg/dose; 2.4 +/- 0.6 mg/kg) than the traditional arm (81 +/- 27 mg/dose; 1.5 +/- 0.6 mg/kg) (p less than .001) . In addition, the dose per day (mg/kg) was greater in the kinetic arm (4.1 +/- 1.5) than the traditional arm (3.2 +/- 1.3) (p less than .001) . The improved survival was achieved by attaining therapeutic peak serum concentrations earlier in the course of the infection and by administering more total aminoglycoside without increasing toxicity . CONCLUSIONS: We conclude that pharmacokinetic management of aminoglycoside dosing may improve the outcome of severely ill patients.

J Bacteriol, 1991 Dec, 173(23), 7684 - 91
Amount of peptidoglycan in cell walls of gram-negative bacteria; Wientjes FB et al.; The amount of diaminopimelic acid (Dap) in the cell wall of Escherichia coli was measured in two ways . A radiochemical method first described by us in 1985 (F . B . Wientjes, E . Pas, P . E . M . Taschner, and C . L . Woldringh, J . Bacteriol . 164:331-337, 1985) is based on the steady-state incorporation of {3H}Dap during several generations . Knowing the cell concentration and the specific activity of the {3H}Dap, one can calculate the number of Dap molecules per sacculus . The second method measures the Dap content chemically in sacculi isolated from a known number of cells . With both methods, a value of 3.5 x 10(6) Dap molecules per sacculus was obtained . Combined with electron microscopic measurements of the surface area of the cells, the data indicate an average surface area per disaccharide unit of ca . 2.5 nm2 . This finding suggests that the peptidoglycan is basically a monolayered structure.

J Bacteriol, 1991 Dec, 173(23), 7534 - 9
Temporal sequence of the recovery of traits during phenotypic curing of a Cytophaga johnsonae motility mutant; Gorski L et al.; The lack of cell translocation and the resulting formation of nonspreading colonies of mutants of the gram-negative gliding bacterium Cytophaga johnsonae have been correlated with the loss of cell surface features of the organism . These cell surface traits include the ability to move polystyrene-latex beads over the cell surface and the ability to be infected by bacteriophages that infect the parent strain . In order to assess whether these traits reflect structures or functions that actually play a role in gliding, we studied a mutant (21A2I) selected for its inability to form spreading colonies; it is deficient in sulfonolipid, lacks bead movement ability, and is resistant to at least one bacteriophage . The provision of cysteate (a specific sulfonolipid precursor) restores lipid content and gliding to the mutant; hence, the lipids are necessary for motility . Growth with cysteate also restores bead movement and phage sensitivity . In order to determine the temporal relationship of these traits, we undertook a kinetic study of the appearance of them after addition of cysteate to the mutant . One predicts that appearance of a trait essential for cell translocation will either precede or accompany the appearance of this ability, while a nonessential trait need not do so . Sulfonolipid synthesis was the only trait that appeared before gliding; this is consistent with its established importance for motility . Bead movement and phage sensitivity first appeared only after gliding started, suggesting that the machinery involved in those processes is not necessary, at least for the initiation of gliding.

J Chemother, 1991 Dec, 3(6), 352 - 6
New epidemiological data on resistance to netilmicin and other aminoglycosides; Toscano MA et al.; In previous investigations we considered the increasing resistance of Gram-negative organisms to aminoglycosides over time . We therefore performed further studies to evaluate eventual variations in the incidence of bacterial resistance to aminoglycosides over the last four years . We tested Gram-negative microorganisms recently isolated from pathological materials of various clinical origins with gentamicin, amikacin, tobramycin and netilmicin . Our data show evidence that there was no significant increasing resistance to aminoglycosides, and that netilmicin continues to be active as an anti-Gram-negative antibiotic.

Mol Microbiol, 1991 Dec, 5(12), 2977 - 81
A protein kinase C-like activity in Escherichia coli; Norris V et al.; The protein kinase C (PKC) family comprises calcium- and phospholipid-dependent kinases whose activity is stimulated by diacylglycerol and tumour-promoting phorbol esters such as 12-tetradecanoyl phorbol-13-acetate (TPA) . In the Gram-negative bacterium Escherichia coli, functional similarity to PKC was demonstrated in crude extracts by calcium and phospholipid-dependent, TPA-stimulated phosphorylation of a small number of endogenous substrates . Activity was reduced by sphingosine, a known inhibitor of eukaryotic PKC . Structural similarity to PKC was demonstrated in crude and partially purified bacterial extracts by cross-reactivity with several monoclonal antibodies . This revealed isozyme-specific homology between a protein(s) of relative molecular mass 80-85,000 in E . coli and the alpha- and gamma-isozymes, but probably not the beta-isozyme, of eukaryotic PKC.

Acta Med Port, 1991 Dec, 4 Suppl 1, 20S - 27S
{Biochemical characterization and metabolic effects of tumor necrosis factor}; Martins e Silva J; The tumor necrosis factor, preliminary identified because of its antitumor properties, refers to two kinds of similar polypeptides (TNF or cachectin, and TNF-beta or lymphotoxin), which share some biological effects . Both substances, as members of the class of cytokines, play a role as mediators of inflammation and the cellular immune response . Human cachectin is produced as a prohormone and activated by cleavage of a 76 residue peptide . Mature cachectin (which comprises 157 amino acid residues) share a 28% amino acid sequence homology with lymphotoxin . Both cytokines are encoded by different genes of chromosome 6 and may compete for a common receptor . Cachectin is produced by a wide variety of cells (phagocytic and non-phagocytic), mainly by activated macrophages and monocytes . Different invasive stimuli (mainly lipopolysaccharide, a constituent of the Gram-negative bacteria's outer wall) activate cachectin biosynthesis, which is controlled chiefly at a post-transcriptional level . The newly synthetized cachectin remains associated as a transmembrane form, affecting their targets by direct cell-to-cell contact, or is actively secreted in the circulation to distant sites in the body, where it binds to high affinity cachectin receptor, on a variety of cell types . Cachectin exerts pleiotropic effects on normal, transformed, or tumoral cells . The biological effects mediate by cachectin may be beneficial or deleterious to the body, depending on the quantity produced, duration of cell exposure and further biochemical mediators in the environment of the target cells . Cachectin (frequently associated with severe infection and cancer) seems to be the result of a persistent exposure to raised levels of cachectin.(ABSTRACT TRUNCATED AT 250 WORDS)

Jpn J Clin Oncol, 1991 Dec, 21(6), 428 - 34
A randomized trial comparing imipenem/cilastatine alone with latamoxef plus tobramycin in febrile neutropenic patients with lung cancer; Matsui K et al.; We conducted a randomized trial to compare the efficacy of imipenem/cilastatine (IPM/CS) monotherapy with that of a combination of latamoxef (LMOX) and tobramycin (TOB) in the initial management of fever and neutropenia in patients with lung cancer . Leukocytopenic febrile patients (less than 3,000 leukocytes per microliters; temperature greater than 38 degrees C) with lung cancer given induction therapy were randomly assigned to receive intravenous treatment with either 1 g IPM/CS twice daily or 2 g LMOX plus 90 mg TOB twice daily . A total 101 febrile episodes were studied . Fifty-one episodes were treated with IPM/CS and 50 with LMOX+TOB . Fifty-nine of the febrile episodes were bacteriologically confirmed, while an organism could not be isolated despite the presence of obvious clinical infection in the remaining 42 . The response rate was 82% with IPM/CS and 80% with combination therapy . This difference was not statistically significant . The response rate regarding gram-negative infections was 10 out of 14 (71%) in the IPM/CS group and seven out of 12 (58%) in the LMOX+TOB group . This difference was also not significant (P = 0.484) . The response rate in severely neutropenic patients (neutrophils less than 100/microliters) was low (P = 0.078) . Three patients in the IPM/CS group were withdrawn from the study due to skin rash and vomiting . Therapy with IPM/CS monotherapy was as effective as a combination regimen.

Lymphokine Cytokine Res, 1991 Dec, 10(6), 487 - 93
In situ activation of mouse lung macrophages by coadministration of liposomes containing the lipopeptide CGP 31362 and interleukin 2 involves interaction with T lymphocytes and natural killer cells; Utsugi T et al.; These studies were undertaken to determine the mechanism for augmented tumoricidal activity of alveolar macrophages (AM) in mice injected intravenously with multilamellar liposomes containing a lipopeptide analogue of Gram-negative bacteria cell wall (MLV-CGP 31362) and intraperitoneally with interleukin 2 (IL-2) . BALB/c mice were injected into the kidney with syngeneic renal carcinoma cells . Ten days later, this kidney was resected, and the mice were treated intravenously with MLV-CGP 31362 and/or intraperitoneally with IL-2 . Treatment with MLV-CGP 31362 led to a reduction in the number of lung metastases, whereas treatment with IL-2 alone did not . The coadministration of intravenous liposomes and intraperitoneal IL-2 produced significant eradication of lung metastases . MLV-CGP 31362 (iv) and IL-2 (ip) were injected both into control immune-competent and nude mice or into mice whose natural killer (NK) cells had been depleted by systemic administration of anti-asialo GM1 antibodies . MLV-CGP 31362 activated tumoricidal properties in AM of all groups of mice . The additive tumoricidal activation of AM by IL-2 was associated with its effects on both T cells and NK cells.

Int J Food Microbiol, 1991 Dec, 14(3-4), 247 - 60
Borrelia burgdorferi: another cause of foodborne illness?
Farrell GM, Marth EH.
Borrelia burgdorferi was identified as the etiological agent of Lyme disease in 1982 . This Gram-negative spirochete is classified in the order Spirochaetales and the family Spirochaetaceae . The pathogen is fastidious, microaerophilic, mesophilic and metabolises glucose through the Embden-Meyerhof pathway . A generation time of 11 to 12 h at 37 degrees C in Barbour-Stoenner-Kelly medium has been reported . Lyme disease, named after Lyme in Connecticut, is distributed globally . It is the most commonly reported vector-borne disease in the United States, where the incidence is highest in the eastern and midwestern states . Since establishment of national surveillance in 1982, there has been a nine-fold increase in the number of cases reported to the U.S . Centers for Disease Control . The deer tick of the genus Ixodes is the primary vector of Lyme borreliosis . The tick may become infected with B . burgdorferi, by feeding on an infected host, at any point in its 2-year life cycle which involves larval, nymphal and adult stages . The infection rate in deer ticks may be as high as 40% in endemic areas . The primary vertebrate reservoirs for Ixodes are the white-footed mouse (Peromyscus leucopus) and the white-tailed deer (Odocopileus virginianus) . Dairy cattle and other food animals can be infected with B . burgdorferi and hence some raw foods of animal origin might be contaminated with the pathogen . Recent findings indicate that the pathogen may be transmitted orally to laboratory animals, without an arthropod vector . Thus, the possibility exists that Lyme disease can be a food infection . In humans, the symptoms of Lyme disease, which manifest themselves days to years after the onset of infection, may involve the skin, cardiac, nervous and/or muscular systems, and so misdiagnosis can occur.

Infect Dis Clin North Am, 1991 Dec, 5(4), 899 - 913
The role of monoclonal antibodies in the management of gram-negative sepsis . Experience with the E5 antibody; Gorelick KJ et al.; Immunologic targeting of the mediators of sepsis is a new approach to reducing mortality associated with this often-fatal complication . When sepsis is due to infection with a gram-negative pathogen, endotoxin plays a key role in its pathogenesis . Antiendotoxin antibody E5 binds endotoxin from a broad spectrum of clinically relevant gram-negative bacteria and reduces mortality from endotoxemia and bacteremia in animal models . It seems to be safe to administer to patients with suspected gram-negative sepsis; fewer than 2% of patients experienced allergic-type reactions, a frequency similar to that seen with third-generation cephalosporins . When administered in a dose of 2 mg/kg daily for two days, E5 reduces mortality and improves the outcome of multi-organ failure in patients with gram-negative sepsis, especially when administered before the development of refractory shock . Patients with sepsis of other etiology have not been shown to benefit from antiendotoxin immunotherapy . E5 antibody appears to be an effective agent for the adjunctive treatment of gram-negative sepsis . Further evaluation of E5 antibody is warranted in the treatment of patients with neutropenia, burns, and shock.

Infect Dis Clin North Am, 1991 Dec, 5(4), 835 - 46
Prevention of infection by gram-negative microorganisms; Verhoef J; The epidemiology and prevention of gram-negative infections are discussed . In addition, the controversial concept of selective decontamination is addressed.

Infect Dis Clin North Am, 1991 Dec, 5(4), 817 - 34
Antibiotic therapy for gram-negative bacteremia; Calandra T et al.; Although antibiotic therapy is the mainstay of therapy for gram-negative bacillary bacteremia, the amelioration of the underlying conditions, the correction of predisposing factors, the drainage of abscesses, the removal of infected foreign bodies, and adequate supportive care are also of paramount importance for curing the infection and should not be neglected . Beginning in the late 1960s, most of the clinical work on gram-negative infections has focused on the evaluation of new antibiotics . Numerous studies have shown that early, appropriate antibiotic treatment of gram-negative bacteremia significantly improved patients' outcomes and prevented the development of septic shock . Prescribing standard doses of antibiotics does not necessarily mean that therapeutic levels will be reached in all patients, and relapses of infections or breakthrough bacteremias can occur in patients with subinhibitory serum levels of antibiotics . The monitoring of serum concentrations of antibiotic is therefore recommended in critically ill septic patients . Whereas initial studies on the antibiotic treatment of gram-negative bacteremia were carried out in nonneutropenic patients, more recent clinical investigations have been performed almost exclusively in cancer patients with neutropenia . Studies conducted in the 1970s and 1980s among these patients have shown the following: (1) early empirical therapy reduced the mortality of gram-negative bacteremia; (2) therapy with a combination of two antibiotics, be it an extended spectrum penicillin plus an aminoglycoside or a third-generation cephalosporin, has significantly improved patients' outcomes; and (3) triple-drug combinations (i.e., a penicillin plus a cephalosporin plus an aminoglycoside) are not superior to combinations of beta-lactams and aminoglycosides . For the treatment of gram-negative bacteremia, clinicians today have a choice between well-established antibiotic combinations and broad-spectrum single-agent therapy with third-generation cephalosporins or carbapenem antibiotics . Although recent studies suggested that monotherapy could be as effective as combination therapy for the empirical treatment of fever in the neutropenic host, no definitive study has so far unquestionably demonstrated the equivalence of these treatments in patients with gram-negative bacteremias, especially those caused by P . aeruginosa, or in patients with adverse prognostic conditions, such as persistent and profound granulocytopenia . This literature should however be reviewed with great caution . Indeed, only a minority of studies have included a sufficient number of patients to confidently assess the impact of therapy on patients' outcomes . Obviously, small studies can have a significant risk of type II errors, that is, making false-negative conclusions.(ABSTRACT TRUNCATED AT 400 WORDS)

Infect Dis Clin North Am, 1991 Dec, 5(4), 781 - 91
Current understanding of the pathogenesis of gram-negative shock; Waage A et al.; There is increasing evidence that gram-negative bacteria via endotoxin induce the excessive production of inflammatory cytokines, which are active in the pathogenesis of septic shock, multiorgan failure, and ARDS . In animals the injection of TNF induces pathophysiologic and histopathologic changes that are characteristic of the septic shock syndrome, and in patients there is a close association between levels of TNF and the severity of the shock . IL-1 and IFN-gamma markedly potentiate the toxic TNF effects in animal experiments . IL-6 is frequently released into serum during septic shock, and its levels are associated with the severity of the shock . The cytokine is probably not directly involved in the pathogenesis of the shock but may contribute to fever, neutrophilia, and production of acute-phase proteins . Endothelial cells and neutrophils are important target cells for the cytokines in mediation of septic shock and late complications . Underlying conditions like cancer, trauma, burns, and other kinds of stress may alter the induction mechanism of the cytokines and the susceptibility of the organism . The pathogenetic significance of TNF and other cytokines in different categories of septic shock remains to be clarified.

Infect Dis Clin North Am, 1991 Dec, 5(4), 739 - 52
Epidemiology and clinical impact of gram-negative sepsis; Martin MA; Gram-negative sepsis, which was uncommon in the pre-antibiotic era, has become a major problem throughout the latter half of this century, particularly among persons with severe underlying diseases . The development of antibiotics with dramatic bactericidal activity against these pathogens and advances in supportive care have improved the outlook . Nonetheless, the mortality remains high among the growing numbers of elderly patients with chronic illnesses, immunocompromised patients, patients subjected to invasive procedures, and patients in critical care units colonized with highly antibiotic-resistant flora.

Onderstepoort J Vet Res, 1991 Dec, 58(4), 227 - 37
The tick-borne rickettsia Cowdria ruminantium has a Chlamydia-like developmental cycle; Jongejan F et al.; The development of the tick-borne rickettsial pathogen Cowdria ruminantium (S stock) was studied in bovine umbilical endothelial (BUE) cell cultures and in goat choroid plexus, by light- and electron microscopy . Cowdria divided by binary fission within intracytoplasmic vacuoles resulting in large colonies of reticulate bodies . After three to four days in culture, reticulate bodies developed into smaller intermediate bodies characterized by an electron-dense core . Shortly before disruption of the host cells, intermediate bodies condensed further into electron-dense elementary bodies, which were released into the culture medium . Elementary bodies invade other endothelial cells thus initiating a new infectious cycle which lasts between 5 and 6 days . In the infected goat choroid plexus similar reticulate and intermediate bodies were identified within vacuoles of capillary endothelial cells . However, extracellular elementary bodies were not detected . Another stock of Cowdria (W) showed an identical developmental cycle as that of the S stock . The W isolate was also pathogenic for mice, making it possible to test the infectivity of reticulate and elementary bodies in these animals . Reticulate bodies appeared to be less infective than elementary bodies . The developmental cycle of Cowdria resembles the cycle known to occur in Chlamydia . Moreover, Cowdria has other similarities with Chlamydia . It has a Gram-negative envelope, it does not store iodine-stainable carbohydrates and may lack peptidoglycan as does Chlamydia . It is concluded, that Cowdria and Chlamydia are to a certain extent related, confirming a recent report that both organisms have certain antigenic determinants in common . Since Cowdria is also related to Ehrlichia it may well be that Cowdria takes an intermediate position between Chlamydia and Ehrlichia . The phylogenetic relationship between Cowdria and Chlamydia and also with Ehrlichia should be further elucidated by molecular analysis using 16S ribosomal DNA sequences.

Microbiol Rev, 1991 Dec, 55(4), 649 - 74
Synthesis of the cell surface during the division cycle of rod-shaped, gram-negative bacteria; Cooper S; When the growth of the gram-negative bacterial cell wall is considered in relation to the synthesis of the other components of the cell, a new understanding of the pattern of wall synthesis emerges . Rather than a switch in synthesis between the side wall and pole, there is a partitioning of synthesis such that the volume of the cell increases exponentially and thus perfectly encloses the exponentially increasing cytoplasm . This allows the density of the cell to remain constant during the division cycle . This model is explored at both the cellular and molecular levels to give a unified description of wall synthesis which has the following components: (i) there is no demonstrable turnover of peptidoglycan during cell growth, (ii) the side wall grows by diffuse intercalation, (iii) pole synthesis starts by some mechanism and is preferentially synthesized compared with side wall, and (iv) the combined side wall and pole syntheses enclose the newly synthesized cytoplasm at a constant cell density . The central role of the surface stress model in wall growth is distinguished from, and preferred to, models that propose cell-cycle-specific signals as triggers of changes in the rate of wall synthesis . The actual rate of wall synthesis during the division cycle is neither exponential nor linear, but is close to exponential when compared with protein synthesis during the division cycle.

Hepatogastroenterology, 1991 Dec, 38(6), 547 - 9
Abdominal sepsis following liver resection in the rat; Andersson R et al.; A standardized 2/3 liver resection was performed in the rat . One and seven days following liver resection, gram-negative sepsis was induced by cecal ligation and puncture (CLP) . Mortality significantly increased following CLP one day after hepatectomy, but no difference was to be seen after seven days as compared with sham operated animals . Clearance of radiolabeled heat-killed 125I E . coli injected intravenously was significantly decreased one day following liver resection, but not after seven days . The capability of bacterial clearance and survival correlated well with the increase in weight of the liver remnant following liver resection, as did organ uptake of radiolabeled bacteria within the liver, spleen and lungs, as a measure of reticuloendothelial system function . Splenic and pulmonary uptake initially increased following liver resection, but normalized within seven days . In conclusion, the present study shows that the liver is responsible for most of the reticuloendothelial system function and that a major liver resection increases the risk of fatal outcome before regeneration of the liver remnant has occurred, despite normal function of the residual liver tissue.

J Gen Microbiol, 1991 Dec, 137 ( Pt 12), 2823 - 30
Identification of single bacterial cells using digoxigenin-labelled, rRNA-targeted oligonucleotides; Zarda B et al.; Oligonucleotides were end-labelled with digoxigenin (DIG), chemically at the 5'-end or enzymically at the 3'-end . Following specific in situ hybridization of these probes to intracellular rRNA molecules, the hybrids were detected with anti-DIG Fab fragments labelled with fluorescent dyes . The antibody fragments penetrated through the bacterial cell periphery and specifically bound to their antigens . Probe-conferred and non-specific fluorescence per cell were quantified by flow cytometry and compared to values obtained with end-labelled fluorescent probes . The DIG reporter molecules could also be detected in whole fixed cells by antibodies labelled with either alkaline phosphatase or horseradish peroxidase . The penetration of the large antibody-enzyme complexes into the cells required lysozyme/EDTA treatment prior to the hybridization and has so far only been achieved for Gram-negative bacteria . This technique has the potential for significant signal amplification as compared to the fluorescently end-labelled oligonucleotides hitherto used for single cell identification in microbial ecology . Moreover, it can be used instead of fluorescent assays in natural samples showing autofluorescence.

Proc Natl Acad Sci U S A, 1991 Dec 1, 88(23), 10586 - 90
Immunoglobulin-like PapD chaperone caps and uncaps interactive surfaces of nascently translocated pilus subunits; Kuehn MJ et al.; Molecular chaperones are found in the cytoplasm of bacteria and in various cellular compartments in eukaryotes to maintain proteins in nonnative conformations that permit their secretion across membranes or assembly into oligomeric structures . Virtually nothing, however, has been reported about a similar requirement for molecular chaperones in the periplasm of Gram-negative bacteria . We used the well-characterized P pilus biogenesis system in Escherichia coli as a model to elucidate the mechanism of action of a periplasmic chaperone, PapD, which is specifically required for P pilus biogenesis . PapD probably associates with at least six P pilus subunits after their secretion across the cytoplasmic membrane, but PapD is not incorporated into the pilus . We used purified periplasmic complex that PapD forms with the PapG adhesin to investigate the function of interactions between the chaperone and its targets . We demonstrated that PapD binds to PapG to form a stable, discrete bimolecular complex and that, unlike cytoplasmic chaperones, the periplasmic PapD chaperone maintained PapG in a native-like conformation . Bound PapD in the complex was displaced by free PapD in vitro; however, the in vivo release of subunits to the nascent pilus is probably driven by an ATP-independent mechanism involving the outer membrane protein PapC . In addition, the binding of PapD to PapG in vitro prevented aggregation of PapG . We propose that the function of PapD and other periplasmic pilus chaperones is to partition newly translocated pilus subunits into assembly-competent complexes and thereby prevent nonproductive aggregation of the subunits in the periplasm . These data provide important information for understanding the mechanism of action of this general class of chaperones that function in the periplasmic space.

Cell, 1991 Nov 29, 67(5), 995 - 1006
A gene encoding a protein serine/threonine kinase is required for normal development of M . xanthus, a gram-negative bacterium; Munoz-Dorado J et al.; PCR reactions were carried out on the genomic DNA of M . xanthus, a soil bacterium capable of differentiation to form fruiting bodies, using oligonucleotides representing highly conserved regions of eukaryotic protein serine/threonine kinases . A gene (pkn1) thus cloned contains an ORF of 693 amino acid residues whose amino-terminal domain shows significant sequence similarity with the catalytic domain of eukaryotic protein serine/threonine kinases . The pkn1 gene was overexpressed in E . coli, and the gene product has been found to be autophosphorylated at both serine and threonine residues . The expression of pkn1 is developmentally regulated to start immediately before spore formation . When pkn1 is deleted, differentiation starts prematurely, resulting in poor spore production . These results indicate that the protein serine/threonine kinase plays an important role in the onset of proper differentiation.

Schweiz Rundsch Med Prax, 1991 Nov 19, 80(47), 1324 - 6
{Cat-scratch disease}; Vassalli G et al.; Cat-scratch disease is usually a self-limited illness characterized by regional lymphadenopathy and a skin inoculation lesion . These findings generally resolve spontaneously over a period of several weeks or months . The discovery of the causative organism, an intracellular gram-negative rod and the recognition of severe systemic courses in immuno-compromised patients have renewed the interest in this disease . We report a prolonged clinical course with lymph node suppuration in an immunocompetent patient . The diagnosis was confirmed by the detection of the organism in Warthin-Starry silver stained lymph node biopsy.

J Cataract Refract Surg, 1991 Nov, 17(6), 847 - 8
Branhamella catarrhalis colonization of exposed nylon sutures; Harris DJ Jr; A 63-year-old man who had uncomplicated cataract surgery four years previously complained of having redness, blurred vision, and photophobia in his right eye for one week . Examination revealed numerous exposed nylon sutures with peculiar leaf-like, white deposits surrounding them, associated with underlying conjunctival leukocytic infiltration . Smears of the adherent material showed gram-negative diplococci and cultures produced heavy growth of Branhamella catarrhalis . Removal of the offending sutures and fortified topical antibiotic therapy resolved the ocular inflammation . The unique form in which this infection presented is a reminder that ocular surgical patients deserve long-term follow-up for problems like exposed sutures.

Vet Pathol, 1991 Nov, 28(6), 492 - 6
Brucella abortus-associated meningitis in aborted bovine fetuses; Hong CB et al.; Granulomatous meningitis was present in 6/33 bovine fetuses from which Brucella abortus (B . abortus) had been isolated . Meningitis was severe in three fetuses, moderate in one fetus, and mild in the remaining two fetuses . The meningitis was characterized by the infiltration of a mixed population of lymphocytes, plasma cells, and macrophages in the leptomeninges . Vasculitis characterized by the infiltration of lymphocytes and plasma cells in the vascular wall was observed in the vessels of the cerebral cortices of 4/6 fetuses . Gram negative coccobacilli were present in the cytoplasm of the leptomeningeal macrophages and extracellularly . Brucellar antigens labeled by the avidin-biotin-peroxidase complex method were present in massive amounts in leptomeningeal macrophages and in small foci of stained cells in the choroid plexus and ependyma . The findings indicate that B . abortus is one of pathogens capable of inducing meningitis in bovine fetuses.

Nucleic Acids Res, 1991 Nov 11, 19(21), 6021 - 5
Specific incorporation of glycine into bacterial lipopolysaccharide . Novel function of specific transfer ribonucleic acids; Gamian A et al.; It has been found that the bacterial endotoxins (lipopolysaccharides, LPSs) contain some amino acids and glycine is the most abundant amino acid in the polysaccharide core preparations of LPSs of gram-negative bacteria . Until now nothing was known about the mechanism of amino acid incorporation into the lipopolysaccharide core . We found that one out of three glycyl-tRNAs(Gly) from Escherichia coli is the donor of amino acid and is the substrate for a putative aminoacyl-tRNA:LPS transferase . We have isolated, purified this tRNA and determined its nucleotide sequence to be major E.coli tRNA(3Gly) . This tRNA(Gly) (anticodon GCC) conserved the tRNA structural features . The assay for determination of the specific incorporation of glycine into the lipopolysaccharide was also invented and described.

J Clin Invest, 1991 Nov, 88(5), 1747 - 54
Dysregulation of in vitro cytokine production by monocytes during sepsis; Munoz C et al.; The production by monocytes of interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF alpha) in intensive care unit (ICU) patients with sepsis syndrome (n = 23) or noninfectious shock (n = 6) is reported . Plasma cytokines, cell-associated cytokines within freshly isolated monocytes and LPS-induced in vitro cytokine production were assessed at admission and at regular intervals during ICU stay . TNF alpha and IL-6 were the most frequently detected circulating cytokines . Despite the fact that IL-1 alpha is the main cytokine found within monocytes upon in vitro activation of cells from healthy individuals, it was very rarely detected within freshly isolated monocytes from septic patients, and levels of cell-associated IL-1 beta were lower than those of TNF alpha . Cell-associated IL-1 beta and TNF alpha were not correlated with corresponding levels in plasma . Upon LPS stimulation, we observed a profound decrease of in vitro IL-1 alpha production by monocytes in all patients, and of IL-1 beta, IL-6, and TNF alpha in septic patients . This reduced LPS-induced production of cytokines was most pronounced in patients with gram-negative infections . Finally, monocytes from survival patients, but not from nonsurvival ones recovered their capacity to produce normal amounts of cytokines upon LPS stimulation . In conclusion, our data indicate an in vivo activation of circulating monocytes during sepsis as well as in noninfectious shock and suggest that complex regulatory mechanisms can downregulate the production of cytokines by monocytes during severe infections.

Paediatr Indones, 1991 Nov-Dec, 31(11-12), 334 - 42
Transfusion in the newborn; Sunarto; Transfusion, either with whole blood or blood components is frequently needed in the neonatal intensive care . Certain aspects are very important to consider . Citrated blood is preferred to heparinized blood . Transfusion must be rational, either with whole blood or blood components . Whole blood is only indicated for repletion of blood volume, exchange transfusion and certain cases in which no blood component needed is available . To improve oxygen carrying capacity, to stop bleeding due to coagulation defect, thrombocytopenic bleeding due to depressed platelets production and to counter gram negative septicemia, blood component is indicated to obtain optimal effects with minimal side effects.

Rozhl Chir, 1991 Nov, 70(10-11), 472 - 9
{Anatomic correction of a double outlet right ventricle using the Taussig-Bing arterial switch}; Hucin B et al.; Four children with double outlet right ventricle type Taussig-Bing were operated in 1988-1990 by anatomical correction-arterial switch . Primary correction of the defect was performed in one infant at the age of three months . Three children had a previous palliative operation (banding of the pulmonary artery, resection of coarctation of the aorta and ductus arteriosus) . Anatomical correction of the defect was performed in a second stage at the age of 14, 15 and 19 months . One child died from Gram-negative septicaemia on the 8th day after operation . Three children are completely free from complaints and develop normally . The authors discuss some technical aspects of the arterial switch operation in infants with double outlet right ventricle.

J Antimicrob Chemother, 1991 Nov, 28(5), 753 - 64
A prospective randomized study comparing once- versus twice-daily amikacin dosing in critically ill adult and paediatric patients; Marik PE et al.; Three hundred and forty eight critically-ill patients with a documented Gram-negative infection were randomized to receive amikacin once- (od) or twice-daily (bd) . The amikacin was given by slow intravenous injection in a daily dose of 20 mg/kg in patients under the age of one year (paediatric group) and 15 mg/kg in patients over the age of one year (adult group) . Paediatric and adult patients on the od regimen received a loading dose of 25 and 20 mg/kg respectively . The dosages were subsequently adjusted to achieve desirable blood levels . Patients received other antibiotics as clinically indicated . Forty-eight patients were withdrawn from the study due to death or azotaemia occurring in the first 72 h . One hundred and fifty five patients (76 paediatric) received an od dose and 145 (65 paediatric) received a bd dose . The clinical cure rate was 83% in the od group compared to 66% in the bd group (P = 0.001) . The bacteriological cure rate was 81% in the od group compared to 58% in the bd group (P = 0.005) . In the paediatric sub-group the cure rate was higher with the od regimen (P = 0.002) but this difference was not statistically significant in the adult patients (P = 0.1) . The serum creatinine rose in 35% of patients in the bd group compared to 21% in the od group (P = 0.05) . Although audiometry was not performed there was no clinical evidence of ototoxicity in any of the patients . In conclusion od amikacin dosing resulted in a higher cure and less nephrotoxicity than conventional bd dosing.

Circ Shock, 1991 Nov, 35(3), 159 - 63
Impaired carnitine transport in the rat heart during E . coli sepsis; Lanza-Jacoby S et al.; To determine the mechanism for the reduced content of myocardial carnitine during gram-negative sepsis, carnitine transport was examined in isolated perfused hearts . Rats were injected i.v . with 8 x 10(7) live colonies of E . coli per 100 g body weight or physiological saline . All rats were fasted after injection for 22 hr to equalize the differences in food intake . Total carnitine uptake in the hearts from both groups was essentially linear between 10 and 20 min of perfusion with 40 microM of carnitine . However, total uptake was significantly reduced by approximately 30% in the hearts from the E . coli-treated rats . In addition to the significant reduction in total carnitine uptake at 40 microM, uptake was significantly lower at 100 microM perfusate carnitine in hearts from E . coli-treated rats . However, total uptake was not significantly different between the groups at 200 and 300 microM perfusate carnitine . Carnitine uptake in the presence of 0.05 M mersalyl acid was comparable in both groups of hearts at all perfusate carnitine concentrations indicating no change in the diffusion component of transport in the hearts from the fasted E . coli-treated rats . The reduction in total uptake at 40 microM and 100 microM of perfusate carnitine was due to a 42% and 26%, respectively, decrease in the carrier-mediated uptake . This data suggests that the reduced content of myocardial carnitine in fasted E . coli-treated rats is due to the decrease in the carrier-mediated component of transport.

Circ Shock, 1991 Nov, 35(3), 139 - 51
High-dose corticosteroid therapy in human septic shock: has the jury reached a correct verdict?
Sjolin J.
Many studies have been published concerning high-dose corticosteroids (HDC) in septic shock . Most of them have been criticized for not adhering to methodologic standards . Four studies were published during the 1980s taking into consideration the criticism of previous trials . The general conclusion following these trials was the HDC should not be used in the treatment of septic shock . The aim of this review is to compare these trials as regards their aims, study design, results, and conclusions . The validity of the data presented is discussed . It is concluded that there is no unequivocal evidence that HDC is beneficial in septic shock, but it must be realized that there are tremendous difficulties in obtaining the true state of affairs in clinical trials regarding patients in septic shock . Accumulated data indicate that HDC, given as one single dose during the initial hours of gram-negative septic shock, more likely than not has a beneficial effect on mortality.

Surv Ophthalmol, 1991 Nov-Dec, 36(3), 223 - 32
Acute anterior uveitis and HLA-B27; Wakefield D et al.; Acute anterior uveitis is a common ocular disease characterized by inflammation of the iris and ciliary body . In the majority of patients presenting with an acute attack of anterior uveitis, the only clues to the pathogenesis of this disease are its close association with the genetic marker HLA-B27 and the likely triggering role of a variety of gram negative bacteria . HLA-B27 acute anterior uveitis appears to be a distinct clinical entity frequently associated with the seronegative arthropathies, such as ankylosing spondylitis and Reiter's syndrome . Recent advances in our understanding of the structure and function of class I HLA molecules have revealed their fundamental function in antigen presentation and this has led to a reevaluation of their role in disease predisposition.

Ther Drug Monit, 1991 Nov, 13(6), 502 - 6
Individualizing amikacin regimens: accurate method to achieve therapeutic concentrations; Zaske DE et al.; Amikacin's pharmacokinetics and dosage requirements were studied in 98 patients receiving treatment for gram-negative infections . A wide interpatient variation in the kinetic parameters of the drug occurred in all patients and in patients who had normal serum creatinine levels or normal creatinine clearance . The half-life ranged from 0.7 to 14.4 h in 74 patients who had normal serum creatinine levels and from 0.7 to 7.2 h in 37 patients who had normal creatinine clearance . The necessary daily dose to obtain therapeutic serum concentrations ranged from 1.25 to 57 mg/kg in patients with normal serum creatinine levels and from 10 to 57 mg/kg in patients with normal creatinine clearance . In four patients (4%), a significant change in baseline serum creatinine level (greater than 0.5 mg/dl) occurred during or after treatment, which may have been amikacin-associated toxicity . Overt ototoxicity occurred in one patient . The method of individualizing dosage regimens provided a clinically useful means of rapidly attaining therapeutic peak and trough serum concentrations.

Nihon Kyobu Shikkan Gakkai Zasshi, 1991 Nov, 29(11), 1499 - 504
{A case of Legionnaires' disease cured with a combination of erythromycin and steroid therapy}; Yonemaru M et al.; A 53-year-old male was admitted to Keio University Hospital with a pneumonia shadow in the left lung field and respiratory failure . Because there was progression of respiratory failure, mechanical ventilation was required to maintain appropriate oxygenation . Although erythromycin administration was started at the time of admission, a steroid (prednisolone 60 mg/day) was added a few days later to temporarily inhibit the acute inflammatory response in the lung parenchyma . This intensive therapy resulted in resolution of the patient's pneumonia and improvement of his respiratory failure . No pathogens were detected in the clinical specimens . Indirect immunofluorescence examination demonstrated a marked increase in titers against Legionella pneumophila serogroup 1, which was sufficient to confirm a diagnosis of Legionnaires' disease . The causative organism of this disease, a gram-negative short rod, is rarely cultured on conventional culture media . Two clinical subtypes are known based on clinical manifestations: 1) the Pontiac fever-type in which the predominant symptom is fever alone; and 2) the pneumonia-type which was observed in the epidemic in Philadelphia when the disease was first reported in 1976 . The present case of Legionnaires' disease was the severe pneumonia-type which was successfully treated with a combination of erythromycin and a steroid.

J Appl Bacteriol, 1991 Nov, 71(5), 439 - 44
Mercuric reductase activity in the adaptation to cationic mercury, phenyl mercuric acetate and multiple antibiotics of a gram-negative population isolated from an aerobic fixed-bed reactor; Henriette C et al.; Eighty-eight strains, isolated from an aerobic fixed-bed reactor and identified to the genus level, were examined for resistance to 21 antibiotics, cationic mercury and phenylmercuric acetate . All except three were able to grow on Mueller-Hinton agar plates containing 8 micrograms/ml mercuric chloride, but only 42 exhibited a mercuric reductase and an organomercurial lyase activity . Furthermore, 82 of them were multiply-antibiotic resistant, whereas no positive correlation between this property and cationic mercury volatilization capacity was found . It was concluded that this bacterial community-adapted response to these selective agents, which has been most often shown to be mediated by R plasmids, was the result of two independent phenomena . Moreover, the high percentage of multiple antibiotic and mercury resistance found in this population suggested that simultaneous selections occurred on filters of bacteria which exhibited mucoid colonies and tolerance to these two categories of stress agents.

Br J Haematol, 1991 Nov, 79(3), 366 - 71
Drug-induced agranulocytosis: prognostic factors in a series of 168 episodes; Julia A et al.; The prognostic value of 36 clinical and analytical parameters at diagnosis in patients with drug-induced agranulocytosis was analysed in an adult population . This multicentre, retrospective study examined possible prognostic factors by multiple logistic regression analysis in a series of 168 clinical episodes . The overall mortality was 16% . Renal insufficiency at diagnosis and the development of bacteraemia were associated with a poor prognosis . Advanced age, decreased leucocyte count, lymphocytopenia, bone marrow myeloid hypoplasia, increased percentage of bone marrow plasma cells and shock were found to be associated with a poor prognosis only in the univariate analysis . An independent analysis of the myeloid cellularity at diagnosis showed an inverse correlation with the time to recovery of the granulocyte counts (r = -0.43; P = 0.001) . Our data indicate that despite some important clinical differences (higher incidence of infections of the oropharynx, shorter period of neutropenia and almost exclusive presence of gram-negative organisms), the infections complicating the treatment of cancer patients have the same prognostic features than those seen in patients with acute agranulocytosis . Therefore the established therapeutic guidelines for neutropenia after cancer chemotherapeutic agents are applicable to patients with acute agranulocytosis.

Surg Neurol, 1991 Nov, 36(5), 343 - 53
Perioperative complications of encephalo-duro-arterio-synangiosis: prevention and treatment; Matsushima Y et al.; We performed encephalo-duro-arterio-synangiosis on 169 sides of 81 Moyamoya and 8 non-Moyamoya patients in the past 10 years . The advantage of this operation is its minimal operative invasion, giving rise to few operative complications . During 10 years of practice, however, we encountered some perioperative problems . Perioperative cerebral infarction of varying severity was seen in six Moyamoya patients . Severe hyperventilation with crying was the main trigger of infarction . Two patients developed wound infection with Gram-negative rods . Removal of an infected bone flap was necessary . One Moyamoya patient developed malignant hyperthermia during the operation . One acute epidural hematoma necessitated an emergency hematoma removal . Five patients showed a temporary aggravation of involuntary movements . Prolonged mild fever and temporary and limited hair loss around the wound were often observed . Cases and their treatment are presented with discussion of preventive measures.

N Engl J Med, 1991 Oct 17, 325(16), 1132 - 6
Helicobacter pylori infection and gastric carcinoma among Japanese Americans in Hawaii; Nomura A et al.; BACKGROUND . Helicobacter pylori are gram-negative spiral bacteria that are associated with chronic gastritis, a known precursor of gastric carcinoma . Persons at high risk for gastric carcinoma have been shown to have a high prevalence of H . pylori infection . METHODS . We studied the relation of H . pylori infection and gastric carcinoma in a cohort of Japanese American men living in Hawaii . The 5908 men were enrolled and examined from 1967 to 1970 . By 1989, 109 cases of pathologically confirmed gastric carcinoma had been identified . The store serum of each patient with gastric carcinoma and of each matched control subject were tested for the presence of serum IgG antibody to H . pylori . RESULTS . Ninety-four percent of the men with gastric carcinoma and 76 percent of the matched control subjects had a positive test for H . pylori antibodies, for an odds ratio of 6.0 (95 percent confidence interval, 2.1 to 17.3) . As the level of antibody to H . pylori increased, there was a progressive increase in the risk of gastric carcinoma (P for trend = 0.0009) . The association was strong even for men in whom the diagnosis was made 10 or more years after the serum sample was obtained (odds ratio, 10.5; 95 percent confidence interval, 2.5 to 44.8) . CONCLUSIONS . Infection with H . pylori is strongly associated with an increased risk of gastric carcinoma . However, most persons infected with H . pylori will never have gastric carcinoma . Therefore, other factors that increase the risk of gastric carcinoma among persons infected with H . pylori need to be identified.

J Biol Chem, 1991 Oct 15, 266(29), 19499 - 509
Macrophage catabolism of lipid A is regulated by endotoxin stimulation; Hampton RY et al.; Lipopolysaccharide (LPS) is a Gram-negative bacterial glycolipid that is believed to cause, by virtue of its stimulatory actions on macrophages and other eukaryotic cells, the life-threatening symptoms associated with Gram-negative infections . Macrophages both respond to and catabolically deactivate LPS . The lipid A moiety of LPS is responsible for the stimulatory actions of LPS on macrophages . We have previously developed methods employing a radiolabeled bioactive lipid A precursor, 4'-32P-lipid IVA, to study the interaction of this class of lipids with animal cells (Hampton, R . Y., Golenbock, D . T., and Raetz, C . R . H . (1988) . J . Biol . Chem . 263, 14802-14807) . In the current work, we have examined the uptake and catabolism of 4'-32P-lipid IVA by the RAW 264.7 cell line in serum-containing medium at physiological temperatures and have studied the effect of LPS stimulation on the ability of these cells to catabolize lipid IVA . RAW 264.7 macrophage-like cells avidly take up 4'-32P-lipid IVA under cell culture conditions at nanomolar concentrations . Uptake of lipid IVA was accompanied by lysosomal dephosphorylation of a fraction of the lipid to yield 4'-monophosphoryl lipid IVA . Chemically generated 4'-monophosphoryl lipid IVA was found to be substantially less bioactive than lipid IVA in the RAW cell, indicating that this catabolic dephosphorylation results in detoxification . In uptake experiments of 3-4 h duration, all metabolism of lipid IVA is blocked by ligands of the macrophage scavenger receptor . In longer experiments (24 h), both scavenger receptor-dependent and -independent uptake are responsible for the lysosomal catabolism of lipid IVA . Preincubation of RAW 264.7 cells with LPS caused dose-dependent inhibition of lipid IVA dephosphorylation . Sufficient LPS stimulation resulted in essentially complete inhibition of lipid IVA catabolism in both short- and long-term uptake experiments . This effect occurred at physiologically relevant concentrations of LPS (IC50 less than 1 ng/ml), and our data indicate that LPS-induced blockade of lipid IVA catabolism was due to the resultant physiological stimulation of the cells, and not inhibition of dephosphorylation by competition for uptake or enzymatic sites or by simple sequestration of labeled lipid IVA by LPS aggregates . We suggest that in the macrophage, LPS can modulate its own catabolism by virtue of its pharmacological properties . This effect of LPS could play a role in LPS pathophysiology as well as in macrophage biology.

Surgery, 1991 Oct, 110(4), 785 - 91; discussion 791-2
Endotoxin filtration and immune stimulation improve survival from gram-negative sepsis; Cheadle WG et al.; Polymyxin B, when bound to a polystyrene fiber (PMX-F), has been used experimentally as an extracorporeal blood filter to reduce serum lipopolysaccharide levels, which are believed to be responsible for physiologic alterations in the septic state . To validate our theory that a combination of PMX-F, systemic antibiotics, and immune stimulation would improve survival, 78 rats were given intravenous doses of Escherichia coli (range, 4.6 to 6.2 X 10(8) colony-forming units/ml) . They were then randomized into groups receiving either systemic gentamicin (n = 10); pretreatment with muramyl dipeptide (n = 11); or extracorporeal hemoperfusion through either a sham column (n = 8), PMX-F-packed column with systemic gentamicin (n = 8); or PMX-F-packed column with systemic gentamicin and muramyl dipeptide pretreatment (n = 8) . Thirty-three control rats received no treatment . Sham hemoperfusion (13%) and control (21%) rats had the lowest survival rate, although increased improvement was noted in rats treated with gentamicin (30%) or the combination of PMX-F filtration and gentamicin (50%) . The most significant improvements occurred in rats pretreated with muramyl dipeptide (53%) and in rats treated with a combination of PMX-F, gentamicin, and muramyl dipeptide (88%) . These data show that lipopolysaccharide filtration and nonspecific immune stimulation are additive to antibiotic therapy and are useful as adjunctive measures in the multimodal treatment of experimental gram-negative bacterial infection.

Infect Control Hosp Epidemiol, 1991 Oct, 12(10), 591 - 6
Surveillance of surgical wound infections following open heart surgery; Sellick JA Jr et al.; OBJECTIVE: To define accurate wound infection rates for the cardiac surgery service based on site of infection and characterization as "deep" or "incisional" and to determine whether a correctable cause for an apparent increase in deep wound infection rates existed . DESIGN: Observational . SETTING: Tertiary-care teaching hospital . PARTICIPANTS: All adults undergoing open heart surgery in 1988 and 1989 . INTERVENTIONS: Changed from razor to clipper preoperative hair removal in January 1989 . RESULTS: Deep sternotomy wound infections decreased significantly from 1.2% in 1988 to 0.2% in 1989 (p = .010) and deep venectomy (vein donor site) wound infections declined from 1.6% to 0.4% (p = .014) during the same time period . Incisional wound infection rates did not change . Patients with deep infections more likely required readmission or operation to treat their infection than those with incisional wound infections . The percentage of gram-negative organisms causing wound infections decreased from 56.3% in 1988 to 34.7% in 1989 (p = .017) . CONCLUSIONS: Preoperative hair removal using a clipper appears to have decreased the risk of deep wound infection compared with razor preparation . The dichotomous wound classification of "deep" and "incisional" distinguished between patients who required additional interventions for treatment of wound infections.

J Biol Chem, 1991 Oct 5, 266(28), 18538 - 42
Purification and functional characterization of MerD . A coregulator of the mercury resistance operon in gram-negative bacteria; Mukhopadhyay D et al.; Mercury resistance operons (mer) from transposons Tn21, Tn501, and plasmid pDU1358 are highly homologous and inducible with Hg2+ . The regulatory gene merR is transcribed from one promoter, which is divergently oriented from the promoter for the other mer genes . MerR, the product of the regulatory gene, negatively regulates its own expression as well as the expression of the other genes . MerR activates transcription of the operon in the presence of inducing concentrations of Hg2+ . The most promoter distal gene, merD, which is cotranscribed with the structural genes, down regulates the mer operon . A frame-shift mutation in merD, created by deletion of 3 bp and an insertion of a 16 bp sequence upstream of the major inverted repeats present at the 3' end of the merD sequence, resulted in increased synthesis of the structural gene transcript and higher level of resistance to Hg2+ by a factor of about 2 . MerD protein was over-produced using a T7 expression system . The overproduced protein was present in the pellet fraction, when cell lysates were centrifuged at a low speed . Approximately 80% pure MerD protein was recovered from the pellet fraction by extracting with a buffer solution containing 5 M urea . The purified protein migrated as a 13,500 molecular weight protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the N-terminal amino acid sequence corresponded to that deduced from the DNA sequence of merD . MerD bound specifically with the mer promoter sequence . DNase I footprinting experiments identified a common mer operator sequence for MerR and MerD.

J Med Microbiol, 1991 Oct, 35(4), 197 - 202
Adherence of Helicobacter pylori to human gastric epithelial cells in vitro; Hemalatha SG et al.; Gram-negative spiral organisms, currently referred to as Helicobacter pylori, are associated with primary gastritis and duodenal ulceration . The organisms colonise gastric mucus and adhere to epithelial cells of inflamed antra . To further examine the binding of H . pylori to human gastric epithelial cells, we developed and characterised an in-vitro bacterial adherence assay . Scanning electronmicroscopy suggested that spiral-shaped bacteria were adherent to the surface of KATO-III cells which were derived from a human gastric adenocarcinoma . Transmission electronmicroscopy confirmed the attachment of H . pylori to these epithelial cells in tissue culture . Some bacteria were adherent to intact microvilli, others were closely adherent to the plasma membrane in regions where microvilli were effaced . In studies with radiolabelled H . pylori, adherence to epithelial cells in tissue culture contrasted with minimal binding of bacteria to polystyrene wells alone . Incubation of bacteria with gastric cells at 4 degrees C significantly reduced adherence of H . pylori . We conclude that adherence of H . pylori to gastric epithelial cells in tissue culture involved "attachment and effacement mechanisms" . This assay could serve as a suitable in-vitro model for the study of the bacterial adhesins and host receptors which mediate attachment of H . pylori to gastric epithelial cell surfaces.

Epidemiol Infect, 1991 Oct, 107(2), 373 - 81
An outbreak of pneumonia and meningitis caused by a previously undescribed gram-negative bacterium in a hot spring spa; Hubert B et al.; An outbreak of infection caused by a previously undescribed Gram-negative bacterium affected people attending a hot (37 degrees C) spring spa in France in 1987 . Thirty-five case of pneumonia and two cases of meningitis occurred . None of these patients died . Attack rates were significantly higher for patients above 70 years old and for male patients . An epidemiological comparison of the 26 hospitalized cases with 52 matched controls suggests that spa treatment early on the first day (OR = 4.8) and attendance at the vapour baths (OR = 10.7) were significant risk factors for acquiring the infection . Person-to-person spread was not thought to have occurred . The same bacterium was isolated from the hot spring water . All strains studied shows a single rRNA gene restriction pattern . Epidemiological data indicated that the thermal water was the source of infection . This outbreak stresses the need for increased surveillance of infections in people attending hot spring spas.

J Am Geriatr Soc, 1991 Oct, 39(10), 979 - 85
A randomized study of ciprofloxacin versus ceftriaxone in the treatment of nursing home-acquired lower respiratory tract infections; Hirata-Dulas CA et al.; OBJECTIVE: To compare the efficacy and safety of ciprofloxacin and ceftriaxone in patients with nursing home-acquired lower respiratory tract infections requiring initial hospitalization . DESIGN: Prospective, randomized trial . SETTING: Extended care nursing homes affiliated with a teaching hospital . PATIENTS: Fifty patients aged 60 years or older with normal or mildly impaired renal function admitted to the hospital for treatment of lower respiratory tract infections . INTERVENTIONS: Twenty-four patients received initial therapy with intravenous ciprofloxacin, 200 mg every 12 hours (19 patients) or 400 mg every 12 hours (5 patients) during the acute phase followed by 750 mg orally every 12 hours during the convalescence phase . Twenty-six patients received initial therapy with intravenous ceftriaxone, 2 g every 24 hours during the acute phase followed by 1 g administered intramuscularly every 24 hours during the convalescent phase . The total duration of therapy was 14 days . MAIN OUTCOME MEASUREMENTS: Successful outcome was defined as resolution or marked improvement in clinical signs and symptoms of lower respiratory tract infection upon completion of the treatment course . RESULTS: Twelve (50%) of the ciprofloxacin-treated and 14 (54%) of ceftriaxone-treated patients had successful outcomes . Recurrent oropharyngeal aspiration was the reason for treatment failure in most patients refractory to either antibiotic . Mortality during therapy was 8% in each group . From 21 satisfactory sputum specimens collected, S . pneumoniae was the most common isolate, followed by H . influenzae and other Gram-negative bacteria . Ciprofloxacin therapy was well tolerated; ceftriaxone therapy was discontinued in two patients (8%) due to adverse reactions (intramuscular pain and drug fever) . CONCLUSIONS: Sequential intravenous/oral ciprofloxacin appears to be as safe and effective as sequential intravenous/intramuscular ceftriaxone . The optimal dosage of intravenous ciprofloxacin in this patient population appears to be 400 mg every 12 hours; however, additional clinical and pharmacokinetic studies with this regimen are warranted.

J Assoc Physicians India, 1991 Oct, 39(10), 747 - 9
Enoxacin therapy for severe pleuro-pulmonary infections; Thadepalli H et al.; Enoxacin, a new 6-fluoroquinolone known to be active in vitro against most common pulmonary pathogens, was evaluated in comparison with ceftazidime, a third generation cephalosporin proven to be effective in the treatment of gram negative pneumonias . Clinical and microbiologic responses to therapy were satisfactory and comparable in both antibiotic groups . Enoxacin could be an effective alternative choice in the treatment of lower respiratory tract infections caused by gram negative organisms.

Hybridoma, 1991 Oct, 10(5), 625 - 31
Generation of monoclonal antibodies against surface antigens of Moraxella bovis; Aleixo JA et al.; Six hybridoma lines producing monoclonal antibodies (MAbs) against Moraxella bovis were established from fusions between the SP2/0 myeloma cells and BALB/c mice splenocytes . Three antibodies were of the IgG1 isotype, two were IgG2a, and one was IgG2b . The specificity of the antibodies was determined by indirect enzyme-linked immunosorbent assay (ELISA) using whole cells of M . bovis and of other Gram-negative bacteria, and lipopolysaccharide (LPS) from M . bovis JUR2 and E . coli as antigens . Ascitic fluid produced by the six hybridoma lines inhibited hemagglutination by M . bovis GF9 . One MAb (35F) reacted specifically with purified M . bovis LPS in the ELISA test . The MAb panel detected heterogeneity among the isolates recovered from different geographical regions.

Neth J Med, 1991 Oct, 39(3-4), 170 - 6
Life-saving immunotherapy with cross-reactive monoclonal antibodies against endotoxin: a critical evaluation of experimental and clinical studies; Cornelissen JJ et al.; In this paper clinical and experimental studies are reviewed for evidence that monoclonal antibodies (MoAbs), directed against common epitopes in the core glycolipid region of lipopolysaccharide (LPS) of gram-negative bacteria, protect against the lethal effect of bacteremia of serologically unrelated gram-negative bacteria . Only those experimental studies that included control MoAbs and excluded the possibility of survival due to endotoxin tolerance, provided evidence for the protective efficacy of cross-reactive MoAbs . Two recently performed clinical trials evaluating the protective value of the MoAbs HA-1A and E5 showed beneficial effects (enhanced survival) by each MoAb in different subgroups of patients with proven gram-negative bacterial infections . It is concluded that immunotherapy by MoAbs is a promising new tool in the treatment of gram-negative bacterial septic shock . However, the pathophysiological mechanism of protection remains to be elucidated and the group of patients which may benefit most remains to be defined more clearly . Therefore, further experimental and clinical studies are warranted.

J Pediatr Surg, 1991 Oct, 26(10), 1156 - 60
Adjuvant effects of beta-adrenergic drugs on indomethacin treatment of newborn canine endotoxic shock; Goto M et al.; Newborns are susceptible to gram-negative sepsis/septic shock, but there is no established method of its treatment . This study was performed to evaluate the adjuvant effects of dopamine and dobutamine in the indomethacin treatment of newborn endotoxic shock . Endotoxic shock was induced in newborn dogs (2 to 10 days old; 300 to 800 g) by Escherichia coli lipopolysaccharide (LPS; 1.5 mg/kg, intravenously {IV}) . Indomethacin (1.5 mg/kg, IV) was injected 5 minutes after LPS injection . Dopamine (5 micrograms/kg/min) or dobutamine (5 micrograms/mg/min) infusion started 5 minutes after LPS injection immediately following indomethacin injection . Hemodynamic parameters were monitored serially for 120 minutes . LPS induced bradycardia and hypotension, decreased the cardiac output and cardiac performance, and increased the total vascular resistance . When dopamine, dobutamine, or indomethacin were used alone, they attenuated the hemodynamic deterioration by LPS . Dopamine infusion following indomethacin administration improved the hemodynamics further, although dobutamine infusion did not . Therefore, we conclude that the adjuvant therapy of dopamine in the indomethacin treatment of newborn endotoxic shock is beneficial.

J Anim Sci, 1991 Oct, 69(10), 4201 - 8
Aquaculture disease and health management; Meyer FP; Disease problems constitute the largest single cause of economic losses in aquaculture . In 1988, channel catfish producers lost over 100 million fish worth nearly $11 million . Estimates for 1989 predict even higher losses . The trout industry reported 1988 losses of over 20 million fish worth over $2.5 million . No data are available on losses sustained by producers of shellfish . Bacterial infections constitute the most important source of disease problems in all the various types of production . Gram-negative bacteria cause epizootics in nearly all cultured species . Fungal diseases constitute the second most important source of losses, especially in the culture of crustaceans and salmon . External protozoan parasites are responsible for the loss of large numbers of fry and fingerling fin fishes and are a cause of epizootics among young shellfish . The number of therapeutants approved by the Food and Drug Administration is limited . Research to support the registration of promising therapeutic agents is urgently needed.

J Antimicrob Chemother, 1991 Oct, 28(4), 523 - 32
Synergic post-antibiotic effect of mecillinam, in combination with other beta-lactam antibiotics in relation to morphology and initial killing; Hanberger H et al.; The synergic in-vitro post-antibiotic effect (PAE) of mecillinam, in combination with either ampicillin, aztreonam, ceftazidime or piperacillin, on a reference strain of Escherichia coli was evaluated by bioluminescence assay of bacterial ATP . Ampicillin, ceftazidime and mecillinam alone induced a concentration dependent PAE (greater than 3 h) on E . coli, whereas aztreonam and piperacillin alone induced a short (less than 1 h) non-dose dependent PAE . At most concentrations, the combination of mecillinam and ampicillin, aztreonam, ceftazidime or piperacillin induced longer PAEs on E . coli than the sum of the individual antibiotics' PAEs . Long PAEs were seen concomitantly with the presence of spheroplasts . In addition to the synergistic PAE, the decrease in colony counts and changes in ATP values after a 2 h exposure to mecillinam, in combination with the other beta-lactam antibiotics, were more prominent than the respective values after exposure to the individual antibiotics . The change in ATP was generally less pronounced than the decrease in colony counts . This could be due to lysis of spheroplasts on agar plates, leading to an over-estimation of the initial killing when assayed by viable counting . Mecillinam, which induced long PAEs on E . coli at almost all concentrations in this study, has a high affinity for penicillin binding protein 2 (PBP 2) and induced spheroplast formation at all concentrations . However, mechanisms other than the affinity for PBP 2 and spheroplast formation are involved in the PAE of beta-lactam antibiotics on Gram-negative bacteria; since the PAE was prolonged when mecillinam was combined with ampicillin, aztreonam, ceftazidime or piperacillin, which bind preferentially to PBP 1 and 3.

J Vet Diagn Invest, 1991 Oct, 3(4), 328 - 33
Light and electron microscopic observations of a segmented filamentous bacterium attached to the mucosa of the terminal ileum of pigs; Sanford SE; Segmented filamentous bacteria were seen attached to apical villous enterocytes of the terminal ileum in 15 of 2,766 live pigs submitted for necropsy over a 6-year period . Infected pigs ranged in age from 2 to 13 weeks . All pigs except 2, however, were greater than 4 weeks old and had been weaned . All infected pigs came from conventional commercial herds with intensive, all-indoor, confinement rearing management systems . The bacteria were gram negative or gram variable and were not associated with any clinical disease . Bacteria were more commonly attached to epithelium on the dome villi in the ileum . Electron microscopic examination revealed organisms composed of a cranial segment with a nipple-like appendage and several other segments making up elongated filaments of various lengths . Each filament was divided into segments by transverse septa . The nipple-like appendage served as an attachment apparatus by indenting the enterocyte surface without physically penetrating it . Occasionally, the colonizing bacterium was itself colonized by small rod-shaped bacteria that completely surrounded the filament.

Bone Marrow Transplant, 1991 Oct, 8(4), 245 - 51
Influence of total body irradiation on infections after autologous bone marrow transplantation; Callum JL et al.; Infectious complications were analysed in 50 consecutive autologous bone marrow transplant (ABMT) patients treated with high dose etoposide and melphalan, 30 of whom also received total body irradiation (TBI) . Fever developed in 44 patients and bacteremia was documented in 13 (26%) . Patients given TBI had increased susceptibility to bacteremia; 11 of 30 patients who received TBI had positive blood cultures, in contrast to two of the 20 who did not (p = 0.035) . In addition, patients who received TBI had significantly more severe diarrhea (p = 0.037) when compared with those who received chemotherapy alone . Thirty-five patients treated with trimethoprim-sulfamethoxazole prophylaxis had a signficantly lower incidence of gram-negative bacteremia (p = 0.024) . However, when those patients who received trimethoprim-sulfamethoxazole until neutrophil recovery were analysed alone, those who were also given TBI still had significantly more bacteremia (p = 0.047) . Forty-seven patients with follow-up of more than 12 months are available for analysis of varicella zoster (VZV) infections . Of the 29 patients who received TBI, 11 (38%) developed VZV infections, in contrast to one of 18 patients (6%) treated with chemotherapy alone (p = 0.013) . These results suggest that addition of TBI to the intensive therapy regimen for ABMT is associated with significantly more bacteremia and late VZV infections.

Zentralbl Bakteriol, 1991 Oct, 275(4), 451 - 66
Capillary gas chromatography of cellular carbohydrates as a means for the differentiation of fastidious, slow-growing or anaerobic gram-negative bacteria--a review; Mutters R; The phenotypic differentiation of microaerophilic or anaerobic Gram-negative bacteria of human origin is traditionally based on the analysis of metabolic characteristics . Due to outstanding growth requirements of many kinds of parasitic bacteria, e.g . of the facultatively and strictly anaerobic members of the human periodontal flora, conventional biochemical tests are time-consuming and sometimes lead to doubtful results . Commercial diagnostic test kits often produce misidentifications because of insufficient databases or inappropriate test combinations . In this situation, cytochemical methods can provide an alternative . Especially in the case of Gram-negative bacteria which contain considerable amounts of carbohydrates in their cell wall lipopolysaccharide layers, capillary gas chromatography of peracetylated carbohydrate derivatives prepared from bacterial whole cell hydrolysates proved to be useful to characterize individual species, and sometimes even subspecies . As soon as a pure culture is available, the derivatization of the cellular carbohydrates to their peracetylated aldononitriles and peracetylated O-methyloximes, respectively, needs only about four hours and the gas chromatographic spectra are easy to evaluate.

Hum Antibodies Hybridomas, 1991 Oct, 2(4), 194 - 9
Biological activity of a human monoclonal antibody to Bordetella pertussis lipooligosaccharide; Brodeur BR et al.; The heterohybridoma cell line HBp2 secreting human monoclonal antibody (hMAb) directed against Bordetella pertussis was generated by fusing SP2/HPT heteromyeloma cells with human spleen lymphocytes, after in vitro stimulation for 6 days . The hybridoma was maintained in culture for more than 1 year with continuous antibody secretion . The hMAb HBp2, an IgM, reacted with untreated and proteinase K-treated B . pertussis outer membrane antigens, whereas the reactivity was lost when the antigen was treated with sodium periodate . Human MAb HBp2 was shown to be specific to B . pertussis LOS by immunoblotting of whole cell extracts after SDS-PAGE . In a dot enzyme immunoassay, HBp2 reacted with all B . pertussis strains and clinical isolates tested except for four atypical variant strains of the LOS B phenotype . Human MAb HBp2 also reacted with a clinical isolate of B . bronchiseptica . No reaction was observed against B . parapertussis and other gram-negative species . Together these studies suggested that HBp2 is reactive with carbohydrate epitopes present on the LOS A . Binding assays with live bacteria demonstrated that hMAb HBp2 reacted with cell surface exposed epitopes on B . pertussis but the antibody did not bind significantly to the surface on intact B . bronchiseptica cells . When examined for bactericidal activity in the presence of complement, hMAb HBp2 showed high lytic capability against B . pertussis while no killing was obtained against B . bronchiseptica . These experiments established that LOS A is a target for human bactericidal antibodies . This antigen merits further investigation as a potentially important component in human immunity to B . pertussis infection.

Curr Opin Genet Dev, 1991 Oct, 1(3), 313 - 8
Biogenesis of the bacterial pilus; Hultgren SJ et al.; The assembly of surface structures in gram-negative bacteria requires specialized secretion and chaperone systems localized on both sides of the cytoplasmic membrane . Major advances have been made over the last year in understanding how these systems form part of a general strategy used by bacteria to cap and target interactive subunits imported into the periplasmic space to outer membrane uncapping and assembly sites.

Curr Opin Biotechnol, 1991 Oct, 2(5), 661 - 7
Protein secretion in bacteria; Gennity JM et al.; Most secretory proteins are synthesized as precursors with an amino-terminal signal peptide . Genetic identification of proteins essential for signal peptide dependent translocation to the Escherichia coli periplasm has led to the biochemical dissection of the secretion pathway . Additional mechanisms exist in Gram-negative bacteria for protein secretion to the extracellular environment.

J Biol Chem, 1991 Sep 15, 266(26), 17358 - 68
Structure and bacterial receptor activity of a human salivary proline-rich glycoprotein; Gillece-Castro BL et al.; Using an overlay technique, we previously showed that the Gram-negative periodontal pathogen Fusobacterium nucleatum binds to a glycoprotein of Mr 89,000 (Prakobphol, A., Murray, P., and Fischer, S.J . (1987) Anal . Biochem . 164, 5-11) in the parotid saliva of some individuals . We now show that deglycosylation of the purified glycoprotein results in loss of receptor activity . Amino acid analysis of the protein core showed predominantly proline, glycine, and glutamic acid/glutamine, a characteristic of proline-rich glycoproteins (PRG) . The amino terminus contained repeating sequences of Ser-Gln-Gly-Pro-Pro-Pro-Arg-Pro-Gly-Lys-Pro-Glu-Gly-Pro-Pro-Pro- Gln-Gly that had significant compositional and sequence homology to that encoded by exon 3 of the PRB3 gene . We analyzed the PRG oligosaccharides by a combination of mass spectrometry techniques and nuclear magnetic resonance spectroscopy . Twenty-seven highly fucosylated structures were identified . The most abundant was as follows (where Fuc is fucose) . (formula; see text) To understand the structural basis of F . nucleatum binding, we screened glycolipids and neoglycolipids carrying carbohydrate structures related to those of the PRG for receptor activity; components with unsubstituted terminal lactosamine residues best supported adherence . Neoglycolipids constructed from PRG oligosaccharides were also receptors . Treatment with beta-galactosidase, but not alpha-fucosidase, abolished binding, suggesting that unsubstituted lactosamine units, including the 6-antenna of the major oligosaccharide, mediate F . nucleatum adherence.

Rev Infect Dis, 1991 Sep-Oct, 13(5), 985 - 92
Use of immunoglobulins in prevention and treatment of infection in critically ill patients: review and critique; Zanetti G et al.; The study of the use of standard intravenous immunoglobulin (IVIG) preparations as adjunctive therapy for seriously ill patients is motivated by the need to restore immunoglobulin G depleted because of trauma or surgery and/or by the need to provide patients with specific antibodies to various microorganisms . Whereas no clinical studies have shown that standard IVIG has therapeutic efficacy, some data suggest that its prophylactic use is beneficial . Antisera or IVIG prepared from individuals who are hyperimmunized with the biologically active, highly conserved core portion of the endotoxin of gram-negative bacteria confer variable degrees of protection in animal models and clinical trials . Two clinical trials with use of monoclonal antibodies to core lipopolysaccharide have been completed . Only subsets of patients with gram-negative sepsis were protected by the monoclonal antibodies, but the results of the studies were discrepant in regard to the specific characteristics of patients who benefited from the administration of these antibodies . Further studies will be necessary to establish whether this therapy can be recommended for critically ill patients.

J Antimicrob Chemother, 1991 Sep, 28(3), 377 - 88
Uptake of a catecholic cephalosporin by the iron transport system of Escherichia coli; Critchley IA et al.; beta-Lactam antibiotics containing a catechol moiety show potent activity against Gram-negative bacteria, particularly organisms grown under iron-limited conditions, suggesting that the iron-regulated outer membrane proteins (IROMPs) play a role in antibiotic uptake . A catecholic C(7) alpha-formamido-substituted cephalosporin showed increased penetration into Escherichia coli cells grown in an iron-deficient medium compared with cells grown in a medium supplemented with iron . In contrast, penetration of the corresponding monohydroxyphenyl analogue was not influenced by iron concentration . Susceptibility studies with mutants of E . coli lacking one or more IROMPs suggested that the catecholic analogue was able to utilize the Fiu (83 kDa) and Cir (74 kDa) proteins, but not the enterobactin receptor FepA (81 kDa) . Mutants lacking both Fiu and Cir showed a specific decreased susceptibility for catechol-containing cephalosporins . Radio-ligand binding studies with a Fe-catecholic cephalosporin confirmed an association with these proteins.

FASEB J, 1991 Sep, 5(12), 2652 - 60
Gram-negative endotoxin: an extraordinary lipid with profound effects on eukaryotic signal transduction; Raetz CR et al.; The lipid A domain of lipopolysaccharide (LPS) is a unique, glucosamine-based phospholipid that makes up the outer monolayer of the outer membrane of most gram-negative bacteria . Because of its profound pharmacological effects on animal cells, especially those of the immune system, lipid A is also known as endotoxin . Despite decades of earlier work, the precise chemistry of endotoxins and the biochemical pathways for their enzymatic synthesis have been elucidated only within the past 5 years . In this review, we summarize the essentials of endotoxin biochemistry and also present recent experiments aimed at identifying surface receptors, signal-transducing elements, transcriptional factors, and key intracellular targets involved in the response of animal cells to endotoxins.

Chest, 1991 Sep, 100(3), 802 - 8
Gram-negative sepsis . Background, clinical features, and intervention; Bone RC; Gram-negative sepsis remains an urgent medical problem, with more than 200,000 cases occurring each year in the United States and an associated mortality rate of 20 to 50 percent . Since the onset of shock greatly worsens prognosis and to encourage early intervention, the term sepsis syndrome was developed to describe the features of a preshock septic state . Early clinical and metabolic indicators are discussed, and current therapy is reviewed . Better understanding of the pathophysiology of endotoxin release from Gram-negative bacteria and advances in biotechnology have led to the development of potential new treatments for sepsis . One such development--monoclonal antibodies to endotoxin--has shown great promise in the effort to block the progression to septic shock, reduce mortality, and decrease the overall costs of sepsis to the patient and to the national economy.

J Bacteriol, 1991 Sep, 173(17), 5585 - 8
Analysis of outer membrane ultrastructure of pathogenic Treponema and Borrelia species by freeze-fracture electron microscopy; Walker EM et al.; We analyzed the outer membrane (OM) ultrastructure of four pathogenic members of the family Spirochaetaceae by freeze fracture . The OM of Treponema pallidum subsp . pertenue contained a low intramembranous particle concentration, indicating that it contains few OM transmembrane proteins . The concave OM fracture faces of Treponema hyodysenteriae and Borrelia burgdorferi contained dense populations of particles, typical of gram-negative organisms . A relatively low concentration of particles which were evenly divided between a small and a large species was present in the concave OM fracture face of Borrelia hermsii; the convex OM fracture face contained only small particles . As for gram-negative bacteria, the convex OM fracture face particle concentrations of these pathogens were low . These spirochetes cleaved preferentially within the OM, in contrast to typical gram-negative bacteria, which tend to fracture within the inner membrane . The OM ultrastructure of T . pallidum subsp . pertenue provides an explanation for the lack of antigenicity of the treponemal surface and may reflect a mechanism by which this pathogen evades the host immune response.

Crit Care Med, 1991 Sep, 19(9), 1104 - 13
Treatment of gram-negative septic shock with an immunoglobulin preparation: a prospective, randomized clinical trial; Schedel I et al.; OBJECTIVE: To evaluate the effectiveness of a polyclonal immunoglobulin (Ig) preparation containing IgG, IgM, and IgA as an adjunctive therapy for septic shock . DESIGN: Prospective, randomized clinical trial . SETTING: A clinical immunology ward at the center for internal medicine in a university hospital . PATIENTS: Fifty-five patients with septic shock were randomly allocated to two groups according to criteria of septic shock . INTERVENTION: One group of patients (n = 27) received a commercially available immunoglobulin preparation (containing high titers of antibodies specific for determinants to bacterial endotoxin) during the first 3 days after inclusion in the study . The other randomized group (n = 28) did not receive any immunoglobulin preparation . MEASUREMENTS AND MAIN RESULTS: During the period of less than or equal to 6 wks after the beginning of clinically apparent septic shock, death related to the septic process occurred in one (4%) of 27 patients who received immunoglobulin . By comparison, nine (32%) of 28 control group patients died during this period (p less than .01) . Within the first 48 hrs after onset of the clinically apparent septic process, significantly increased activity of circulating endotoxin and simultaneously decreased specific IgG serum titers to lipid A were detected in the group of nonsurvivors . CONCLUSION: Administration of a polyclonal immunoglobulin preparation in the early phase of septic shock was associated with significantly improved survival.

Infect Immun, 1991 Sep, 59(9), 3333 - 6
Distribution, expression, and long-range mapping of legiolysin gene (lly)-specific DNA sequences in legionellae; Bender L et al.; The legiolysin gene (lly) cloned from Legionella pneumophila Philadelphia 1 confers the phenotypes of hemolysis and browning of the culture medium . An internal lly-specific DNA probe was used in Southern hybridizations for the detection of lly-specific DNA in the genomes of legionellae and other gram-negative pathogenic bacteria . Under conditions of high stringency, the lly DNA probe specifically reacted with DNA fragments from L . pneumophila isolates; by reducing stringency, hybridization was also observed for all other Legionella strains tested . No hybridization occurred with DNAs isolated from bacteria of other genera . The lly gene was mapped by pulsed-field gel electrophoresis to the respective genomic NotI fragments of Legionella isolates . By using antilegiolysin monospecific polyclonal antibodies in Western blots (immunoblots), Lly proteins could be detected only in L . pneumophila isolates.

J Infect Dis, 1991 Sep, 164(3), 599 - 601
Comparison of the early dynamics of systemic prostacyclin release after administration of tumor necrosis factor and endotoxin to healthy humans; van der Poll T et al.; Excessive production of prostaglandins may be of importance for the development of organ damage in generalized infection . To investigate the role of tumor necrosis factor (TNF) in systemic prostacyclin release in gram-negative septicemia, the plasma concentrations of its stable metabolite 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) were sequentially measured after intravenous bolus injections of recombinant human TNF (50 micrograms/m2; n = 6) and Escherichia coli endotoxin (2 ng/kg; n = 3) in healthy men . TNF induced a rapid increase in plasma 6-keto-PGF1 alpha from 0.11 +/- 0.01 to 0.44 +/- 0.15 ng/ml after 30 min (P less than .001) . Endotoxin also elicited a rise in plasma 6-keto-PGF1 alpha, but peak values were reached only after 90 min (from 0.07 +/- 0.01 to 0.19 +/- 0.04 ng/ml; P less than .002) . These results indicate that TNF may serve as an intermediate factor in systemic elaboration of prostacyclin in endotoxemia and gram-negative septicemia.

J Clin Microbiol, 1991 Sep, 29(9), 2041 - 2
Meningitis caused by Psychrobacter immobilis in an infant; Lloyd-Puryear M et al.; Psychrobacter immobilis was isolated from the cerebrospinal fluid (CSF) and blood of a 2-day-old infant who appeared well except for a fever and a full anterior fontanelle . The infant was treated with antibiotics intravenously . After 48 h, he became afebrile and CSF and blood cultures were negative; he was then discharged . After 96 h of incubation, CSF and blood cultures yielded a gram-negative organism, P . immobilis . The child was readmitted to the hospital, and the same organism was again isolated from his blood and CSF.

Circ Shock, 1991 Sep, 35(1), 60 - 4
Prophylaxis and treatment of newborn endotoxic shock with anti-lipid A monoclonal antibodies; Goto M et al.; The newborn is very susceptible to gram-negative sepsis/septic shock . The mortality of newborn endotoxic shock continues to be high . Since lipid A is responsible for the toxic effects of lipopolysaccharide, anti-lipid A antibodies may prevent endotoxic shock in the newborn . This study showed that both anti-lipid A monoclonal IgG (A78S1) and anti-lipid A monoclonal IgM (A523) decreased the mortality of endotoxic shock in 10 day old rats . Prophylactic administration of A78S1 and A523 to the pregnant rat decreased the mortality of endotoxic shock in their 0-day-old offspring . Prophylaxis was due to transplacental passage of A78S1 treatment . The mechanism of prophylaxis remains unclear in A523 treatment.

JPEN J Parenter Enteral Nutr, 1991 Sep-Oct, 15(5), 503 - 8
Harry M . Vars Research Award . Arginine supplementation improves histone and acute-phase protein synthesis during gram-negative sepsis in the rat; Leon P et al.; Mechanisms of nutrient alteration of hepatic protein synthesis during sepsis are unclear . In vitro, arginine downregulates endotoxin-stimulated hepatocyte protein synthesis but in vivo effects are unknown . This study evaluated the effects of supplemental arginine or glycine on fibrinogen (acute-phase protein), histone, albumin, and liver protein synthesis after Gram-negative sepsis in the rat . Adult rats (225 g, n = 36) were randomized to receive isonitrogenous isocaloric total parenteral nutrition supplemented with 264 mg of N per kilogram per day as either arginine or glycine . On day 5, each group was further randomized to control or sepsis . Sepsis was induced by injection of 8 x 10(7) Escherichia coli per 100 g body weight, and then a continuous infusion of {1-14C}leucine was started . The rats were sacrificed 4 hours later . The fractional protein synthesis rates (percent per day) of histone, fibrinogen, albumin, and liver were determined . Supplemental arginine led to significantly increased histone (p less than 0.05, analysis of variance) and fibrinogen (p less than 0.01, analysis of variance) synthesis in the septic rats compared with all other groups . Histone and albumin synthesis were also significantly increased (p less than 0.05) in the arginine-supplemented control group compared with the glycine-supplemented control group . Arginine supplementation during sepsis significantly increased (p less than 0.05) albumin and liver protein synthesis compared with controls . Histones which are involved in DNA synthesis and are rich in arginine may play a role in the host response to stress and sepsis . These in vivo results appear to contradict hepatocyte-Kupffer cell coculture studies perhaps because of the hormonal and cytokine responses to nutrient substrate and acute septicemia.

J Hosp Infect, 1991 Sep, 19 Suppl C, 7 - 17
The ecology and immunology of the gastrointestinal tract in health and critical illness; Marshall JC; Interactions between the indigenous flora of the gastrointestinal tract and immunologically competent cells in the gut mucosa, gut-associated lymphoid tissues and liver play an important role in normal immune homeostasis . The microbial flora of the normal gut is complex, yet remarkably constant over time . The relative sterility of the upper gut is maintained by multiple factors including gastric acid, bile salts, normal motility and mucosal IgA, while the lower gut is densely colonized with a complex flora . An intact Gram-negative flora is a prerequisite for normal immunological maturation . On the other hand, overgrowth of the gut, particularly by Gram-negative bacteria or fungi, facilitates the translocation of bacteria into the host, and results in suppression of T-cell responses and altered hepatic Kupffer cell function . Bacterial overgrowth and the consequences of the interactions of this potentially pathogenic flora with the gut immune system may contribute to the septic state in critical illness, and to the syndrome of multiple organ failure.

JAMA, 1991 Aug 28, 266(8), 1097 - 102
A controlled clinical trial of E5 murine monoclonal IgM antibody to endotoxin in the treatment of gram-negative sepsis . The XOMA Sepsis Study Group; Greenman RL et al.; OBJECTIVE: To assess the efficacy of adjunctive monoclonal antibody antiendotoxin immunotherapy in patients with gram-negative sepsis . DESIGN: Double-blind, randomized, placebo-controlled trial . SETTING: Thirty-three university-affiliated centers, including Veterans Affairs, community, and municipal hospitals . PATIENTS: Hospitalized adults with signs of gram-negative infection and a systemic septic response . INTERVENTION: Patients were assigned to receive either 2 mg/kg of a murine monoclonal antibody directed against gram-negative endotoxin (E5) or placebo . A second infusion was administered 24 hours later . MAIN OUTCOME MEASURES: Mortality over the 30-day study period, resolution of organ failures, and safety . RESULTS: Four hundred eighty-six patients were enrolled . Three hundred sixteen had confirmed gram-negative sepsis (54% bacteremic, 46% nonbacteremic) . The survival difference was not statistically significant for all patients . Among patients with gram-negative sepsis who were not in shock at study entry (n = 137), E5 treatment resulted in significantly greater survival (relative risk, 2.3; P = .01) . Resolution of individual organ failures was more frequent among these patients, occurring in 19 (54%) of 35 patients in the E5 group vs eight (30%) of 27 in the placebo group (P = .05) . Four reversible allergic reactions occurred among 247 patients (1.6%) receiving E5 . No other toxicity was identified . CONCLUSIONS: Treatment with E5 antiendotoxin antibody appears safe . It reduces mortality and enhances the resolution of organ failure among patients with gram-negative sepsis who are not in shock when treated.

Biochemistry, 1991 Aug 27, 30(34), 8415 - 23
Expression and characterization of recombinant human acyloxyacyl hydrolase, a leukocyte enzyme that deacylates bacterial lipopolysaccharides; Hagen FS et al.; The molecular cloning and eukaryotic cell expression of the complementary DNA for human neutrophil acyloxyacyl hydrolase (AOAH) are described . AOAH is a leukocyte enzyme that selectively removes the secondary (acyloxyacyl-linked) fatty acyl chains from the lipid A region of bacterial lipopolysaccharides (endotoxins), thereby detoxifying the molecules . The two disulfide-linked subunits of the enzyme are encoded by a single mRNA . The amino acid sequence of the protein contains a lipase consensus sequence in the large subunit and a region in the small subunit that is similar to the saposins, cofactors for sphingolipid hydrolases . The recombinant enzyme, like native AOAH, hydrolyzes secondary acyl chains from more than one position on the lipopolysaccharide backbone . Acyloxyacyl hydrolase is a novel two-component lipase that, by deacylating lipopolysaccharides, may modulate host inflammatory responses to Gram-negative bacterial invasion.

Curr Opin Oncol, 1991 Aug, 3(4), 628 - 33
Use of blood and blood products for supportive care in cancer treatment; Wallerstein R Jr et al.; Prophylactic platelet transfusion can prevent spontaneous hemorrhage in thrombocytopenic cancer patients undergoing intensive treatment, but it is limited by the development of platelet alloimmunization . Exclusive use of leukocyte-depleted blood products delays or prevents the development of platelet alloimmunization and also decreases the frequency of chill and fever reactions and reduces the transmission of cytomegalovirus and possibly other viral infections . Management of disease refractory to platelet transfusion remains a difficult problem . Intravenous immune globulin reduces the incidence of infection and may modify the severity of gram-negative sepsis, decreases the incidence of acute graft-versus-host disease, and may benefit some patients with platelet refractoriness . All blood products given to severely immunocompromised patients, including those undergoing chemoradiotherapy and high-dose therapies, should be irradiated . In addition, designated donor transfusions from first-degree relatives (even when given to immunocompetent patients) should be irradiated.

Gastroenterology, 1991 Aug, 101(2), 437 - 45
Helicobacter pylori infection induces antibodies cross-reacting with human gastric mucosa; Negrini R et al.; The authors' previous observation that many of the monoclonal antibodies against Helicobacter pylori cross-react with the cells of the human gastric mucosa prompted them to investigate the possibility that gastric self-antigens cross-reacting with H . pylori could be involved in the immune response against this organism . It was found that three antibodies against H . pylori, CB-4, CB-10, and CB-14, that cross-react with the human gastric mucosa also intensely cross-reacted with murine gastric epithelial cells . A strong reaction against autologous mucosa was also evident in the sera of mice immunized with H . pylori but not with other bacteria . A serological study performed in a group of 82 patients undergoing gastroscopy showed that the presence of seropositivity against H . pylori was strongly correlated with the presence of autoantibodies against human antral gastric mucosa . This activity was neutralized after absorption of the sera with H . pylori but not with other gram-negative bacteria . The antibodies in the mouse and in the human did not react with other segments of the gastrointestinal tract or with most of the other organs . Mice bearing hybridomas secreting a cross-reacting antibody (CB-4) had histopathologic abnormalities in their stomachs . These lesions were absent in the stomachs of mice bearing hybridomas secreting a non-cross-reacting antibody (CB-26) . It was concluded that H . pylori infection can stimulate antibodies cross-reacting with gastric autoantigens and that this immunologic mechanism may represent a pathogenic link between H . pylori and gastritis.

Microb Pathog, 1991 Aug, 11(2), 111 - 21
Structure and function of the B and D genes of the Actinobacillus actinomycetemcomitans leukotoxin complex; Lally ET et al.; The Actinobacillus actinomycetemcomitans leukotoxin gene complex, consisting of four genes, has been cloned and the sequence of the AaLtC and AaLtA genes reported . The present paper details the sequences of the AaLtB and AaLtD genes which, like AaLtC and AaLTA, are also homologues of genes found in other cytolytic toxin complexes of several other Gram-negative bacterial pathogens . When tested in a recombinant expression system, the AaLtB and/or AaLtD genes are required for the translocation and insertion of the A . actinomycetemcomitans leukotoxin (AaLtA) into the cell membrane of Escherichia coli.

Aust N Z J Ophthalmol, 1991 Aug, 19(3), 217 - 20
Kingella kingae endophthalmitis in an infant; Carden SM et al.; An 11-month-old girl presented with metastatic endophthalmitis . The causative organism was the Gram-negative coccobacillus Kingella kingae . This is an uncommon organism first recognised in 1967 . It has never been previously reported to cause endophthalmitis, but is associated with at least four other infective syndromes in children . Endophthalmitis is a potentially lethal and sight-threatening disease . Kingella kingae is primarily a paediatric pathogen which fortunately responds well to antibiotics . A case study, details of the bacteriology and a table of other clinical syndromes associated with this organism seen at our institution constitute this article . (12 references).

J Immunother, 1991 Aug, 10(4), 236 - 46
Optimization of the liposomes encapsulating a new lipopeptide CGP 31362 for efficient activation of tumoricidal properties in monocytes and macrophages; Nii A et al.; The purpose of this study was to optimize a suitable liposomal carrier for CGP 31362, a new synthetic lipopeptide analogue of gram-negative bacterial cell walls . CGP 31362 was inserted into the membranes of different liposomes with different phospholipid composition . We determined the ability of these liposomes to activate tumoricidal properties in mouse peritoneal and bone marrow macrophages, and in human monocytes . The ideal liposome carrier for CGP 31362 consisted of phosphatidylcholine and phosphatidylserine in a 7:3 molar ratio . Subsequent to efficient binding and endocytosis, CGP 31362 in liposomes of this composition rendered mouse macrophages and human monocytes highly tumoricidal . Moreover, even in the absence of interferon-gamma, human monocytes released significant levels of tumor necrosis factor and interleukin-1 . These data show that in a suitable liposomal carrier, the new synthetic lipopeptide in liposomes is a potent activator of tumoricidal properties in macrophages.

J Surg Res, 1991 Aug, 51(2), 106 - 12
Polymyxin B reduces cecal flora, TNF production and hepatic steatosis during total parenteral nutrition in the rat; Pappo I et al.; Hepatic complications are common in patients receiving total parental nutrition (TPN) and who have no underlying liver disease . In the present study we examined the hypothesis that endotoxin (LPS) or possibly TNF derived from the overgrowth of intestinal gram-negative bacteria is responsible for TPN-associated hepatic steatosis, and that bowel decontamination and specific anti-LPS activity of polymyxin B will reduce fatty infiltration of the liver during TPN . Forty-five male Sabra rats underwent jugular vein cannulation, were placed in metabolic cages, and were randomized into five groups . Group I was continuously infused with normal saline and allowed food ad lib, while group II-V were continuously infused with a TPN formula containing 4.25% amino acids and 25% dextrose for a total of 36 calories and 3.0 g protein per 100 g body weight/day . In addition, groups III-IV were also treated by oral polymyxin B while Groups IV and V received a combination of neomycin, metronidazole, and vancomycin (NMV) . Thus, Group III received polymyxin B, Group IV received both polymyxin B and NMV, while Group V received NMV only . On Days 7-8 of the study, all animals were sacrificed and spontaneous production of TNF by peritoneal macrophages, bacterial translocation to mesenteric lymph nodes, culture of the cecum, and fat, triglyceride, and cholesterol contents of the liver were determined . All groups infused with TPN exhibited higher levels of total fat, triglycerides, and cholesterol compared to the free feeding control group (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

J Trauma, 1991 Aug, 31(8), 1083 - 6; discussion 1086-7
Bacterial translocation in trauma patients; Peitzman AB et al.; Sepsis and multiple system organ failure (MSOF) are major causes of morbidity and mortality in trauma patients . Bacterial translocation induced by hypotension, endotoxemia, or burns is a reproducible phenomenon in the laboratory . The incidence of bacterial translocation to mesenteric lymph nodes (MLNs) in 29 critically ill patients was evaluated to determine its relationship to subsequent sepsis and MSOF . Bacterial translocation was documented in 3 of 4 patients who underwent laparotomy for gastrointestinal (GI) disease . No trauma patient (25 patients), even at second exploration 3-5 days after injury, had a positive MLN culture . Five patients died; 4 trauma patients, one with GI disease . Forty percent of the trauma patients had major complications, predominantly pulmonary infections with gram-negative bacteria . However, infectious complications and outcome were not related to MLN culture results . The classical progression of bacteria from the gut to the bloodstream via the MLNs may require time and gut mucosal injury . The data suggest that bacterial translocation to the MLNs is not a common occurrence in acutely injured trauma patients.

Clin Chem, 1991 Aug, 37(8), 1415 - 9
Computerized approach to monitoring aminoglycosides; Leal T et al.; Aminoglycosides are still used extensively in the treatment of nosocomial infections with Gram-negative bacteria . However, the treatment is associated with several adverse effects . Aminoglycosides monitoring is therefore essential to prevent toxic accumulations and to reach therapeutic concentrations . A computer program, PHARMONITOR, has been developed to optimize aminoglycosides monitoring, responding to the demands of most clinical daily situations . This program, based on a one-compartment open pharmacokinetics model, is developed for IBM PC-compatible computers, using D-Base III+ . It can calculate t1/2, Vd, Cldrug, Cpmax, and the theoretical optimal dose and interval and also evaluates the creatinine clearance . The program has been conceived to allow maximal speed, flexibility, and reliability by the use of (e.g.) a linear least-squares analysis, the possible reference to previous protocols, the extensive use of keywords to classify and recall patients according to their pathologies, the development of messages recommending maximal dose or minimal dosing interval, and increasing the safety of the analysis . We consider the program a valuable tool for adjusting aminoglycoside dosage in individuals.

Mol Gen Mikrobiol Virusol, 1991 Aug, (8), 20 - 2
{Cloning of the REP-region of plasmids from a broad circle of pM3 hosts (INC P-9)}; Titok MA et al.; The rep-region of a broad host range plasmid pM3 (IncP-9) has been cloned by connecting the 7.2 kb PstI fragment of the plasmid with the 1.4 kb PstI fragment of the pUC4K vector plasmid . The latter includes the gene for kanamycin resistance and is unable to replicate by itself . The obtained minireplicon designated pMT2 contains rep-genes permitting its inheritance in the different gram-negative cells and a mob-site coding for incompatibility and temperature instability characteristic of the parent plasmid pM3.

J Clin Microbiol, 1991 Aug, 29(8), 1589 - 92
Clinical and microbiological observations on CDC group DF-3, a gram-negative coccobacillus; Gill VJ et al.; Sequential stool cultures submitted for routine culture were screened for the presence of CDC group DF-3 . Of 690 specimens, 11 (1.6%) yielded moderate to heavy growth of DF-3 . Information on the 11 patients from whom these specimens were obtained showed that 4 had a history of prolonged diarrheal disease that resolved after specific therapy to eradicate DF-3, while for the other 7 patients no clear role could be established . Microbiological characterization of the stool isolates and 10 CDC strains of DF-3 suggested the presence of two subtypes within the group . Antibiotic susceptibility studies showed DF-3 to be relatively resistant to a wide variety of antibiotics.

J Pharm Pharmacol, 1991 Aug, 43(8), 589 - 91
Cepharanthine (biscoclaurine alkaloid) treatment in endotoxic shock of suckling rats; Goto M et al.; Gram-negative sepsis/septic shock causes significant mortality in newborns . However, there has been no established method for newborn endotoxic shock treatment . Prostaglandins play a role in endotoxic shock . Cepharanthine is a biscoclaurine alkaloid that primarily inhibits phospholipase A2 . Therefore, the effects of cepharanthine have been studied on endotoxic shock in newborn rats . Cepharanthine decreased the 24 h mortality of endotoxic shock in a dose-related manner . At the dose of 0.2 mg kg-1 it effectively reduced the mortality from 90 to 21% in newborn rats . It also induced hyperglycaemia in control rats and blunted the hypoglycaemia of endotoxic shock . Cepharanthine did not suppress body weight gain nor did it delay death as seen with glucocorticoid treatment . We conclude that cepharanthine is beneficial in the treatment of newborn endotoxic shock.

Immunology, 1991 Aug, 73(4), 428 - 32
Lipopolysaccharide augments HLA-A,B,C molecule expression but inhibits interferon-gamma-induced HLA-DR molecule expression on cultured human endothelial cells; Otsuka A et al.; The effect of bacterial lipopolysaccharide (LPS) on the expression of class I and II major histocompatibility complex (MHC) molecules on the surface of cultured human umbilical vein endothelial cells (HUVEC) was determined by indirect immunofluorescent staining followed by flow cytometric analysis . LPS at concentrations higher than 0.01 micrograms/ml augmented class I MHC (HLA-A,B,C) expression on HUVEC in a concentration-dependent manner . Optimal augmentation, approximately sixfold compared with control, was seen with 10 micrograms/ml of LPS . Time-course experiments indicated that the augmentation was maximal on Day 4 . In contrast, LPS had no effect on the induction of class II MHC (HLA-DR) molecules and at concentrations higher than 0.01 micrograms/ml inhibited the interferon-gamma(IFN-gamma)-induced class II MHC expression . The inhibition was about 60% at the concentration of 100 micrograms/ml of LPS . Interleukin-1 (IL-1) had a similar effect as LPS on class I and II MHC expression . However, LPS appeared to affect MHC expression directly and not through production of IL-1 or cyclo-oxygenase pathway products, since anti-IL-1 antibodies or an inhibitor of cyclo-oxygenase pathway products, indomethacin, failed to reverse the effects of LPS . These data stress the role of LPS as a direct modulatory factor of class I and II MHC expression on endothelial cells during the development of immune and inflammatory response against Gram-negative bacteria.

Appl Microbiol Biotechnol, 1991 Aug, 35(5), 631 - 7
Expression of the core antigen gene of hepatitis B virus (HBV) in Acetobacter methanolicus using broad-host-range vectors; Schroder R et al.; Using the broad-host-range promoter probe vector pRS201 for cloning of phage Acm1 promoters, we established a convenient vector system for expression of heterologous genes in different Gram-negative bacteria . The usefulness of this system was demonstrated by expression of the HBV core gene in Acetobacter methanolicus . Plasmids carrying the HBV core gene downstream of different Acm1-phage promoters were transferred to A . methanolicus, a new potential host for recombinant DNA expression . Using enzyme immunoassay and immunoblot techniques, the amount and composition of core antigen produced in A . methanolicus were compared with that derived from Escherichia coli . The expression of immunoreactive core antigen in A . methanolicus exceeds by sevenfold that in E . coli using an expression system with tandemly arranged promoters . Morphological observations by electron microscopy show that the HBV core gene products isolated from both hosts are assembled into regular spherical particles with a diameter of about 28 nm that are comparable to original viral nucleocapsids.

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