Gram negative

Infect Immun, 1985 Oct, 50(1), 123 - 9
Sensitivity of Capnocytophaga species to bactericidal properties of human serum; Wilson ME et al.; Capnocytophaga is a newly described genus of gram-negative bacteria which can cause serious oral and extraoral infections in the susceptible host . In the present study, sensitivity of Capnocytophaga spp . to the bactericidal properties of human serum was investigated . Laboratory strains representative of Capnocytophaga sputigena, C . ochracea, and C . gingivalis and seven oral isolates of Capnocytophaga spp . obtained in primary culture were determined to be sensitive to killing by pooled normal serum . In contrast, little or no killing of Capnocytophaga spp . was observed when these organisms were incubated in the presence of hypogammaglobulinemic serum despite evidence for alternative pathway activation . However, hypogammaglobulinemic serum could be reconstituted to bactericidal activity by the addition of the immunoglobulin M fraction of normal serum . Capnocytophaga spp . failed to activate the classical pathway in hypogammaglobulinemic serum, thus ruling out an antibody-independent mechanism of killing . In contrast, good correlation was observed between serum-mediated killing and antibody-dependent classical pathway consumption . These results indicate that complement in the presence of bactericidal antibody may be an important determinant of host resistance to intra- and extraoral infections caused by Capnocytophaga spp.

Tijdschr Kindergeneeskd, 1985 Oct, 53(5), 185 - 8
{Neonatal meningitis: is the combination ampicillin-gentamycin obsolete?}; Muytjens HL et al.; The course of E . coli-meningitis in two infants, treated with latamoxef (moxalactam) in combination with ampicillin is described . Because of the disappointing results the potential value of latamoxef for therapy of Gram-negative enteric meningitis of infancy is discussed.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1985 Oct, 260(2), 184 - 96
Membrane proteins of Legionellaceae . II . Serogroup- and species-specific antigens in the outer membrane of Legionella pneumophila; Ehret W et al.; Antigens of the outer membrane of Legionella pneumophila were investigated by means of the immunoblotting-technique using rabbit antisera against three different formaldehyde-inactivated strains, and one heat-inactivated strain of L . pneumophila serogroup 1 . Nitrocellulose blots were prepared from membrane fractions extracted with sodium-N-lauryl-sarcosinate from 14 strains of L . pneumophila (eight strains of serogroup 1, and one strain each of serogroups 2-7) and 12 strains of gram-negative rods of various species . After incubation with 125I-protein A or 125I-anti-rabbit IgG immune complexes were identified . These results were compared with Coomassie-stained and silver-stained SDS gels . There was a diffuse reaction in the homologous system between 20 and 80 kilodalton (kDal) after incubation with 125I-protein A, and an intense reaction between 22 and 29 kDal after incubation with 125I-anti-rabbit IgG . Membrane preparations of the different strains of serogroup 1 exhibited clearly discernible patterns . Immunoblots of formaldehyde-inactivated strains when reacted with antiserum against heat-inactivated immunogen showed a single species-specific antigen of approximately 22.5 kDal which could not be assigned to a major protein . Immunoblots of the same antiserum but with heat-inactivated cell wall preparations gave a second species-specific band of approximately 65 kDal . Antisera against formaldehyde-inactivated bacteria demonstrated more complex characteristic patterns, with protein-associated components occurring at 29, 44, 46, 48, 65 and 80 kDal; in addition, cross-reacting fractions were present at 15.5, 17.5 and 22.5 kDal . The 29 kDal major outer membrane protein was immunogenic in most but not all cases.

Arch Intern Med . 1985 Oct;145(10):1908.
Escherichia coli bacteremia, meningitis, and hemochromatosis; Christopher GW; A 67-year-old man with Escherichia coli bacteremia and meningitis was found to have hemochromatosis . To my knowledge this is the first documented case of E coli meningitis occurring in the setting of hemochromatosis . The case raises issues regarding the role of chronic liver disease in the pathogenesis of gram-negative sepsis and the impact of iron loading on host immunocompetence and bacterial virulence.

J Infect Dis, 1985 Oct, 152(4), 811 - 6
Efficacy of ciprofloxacin in experimental arthritis caused by Escherichia coli--in vitro-in vivo correlations; Bayer AS et al.; Ciprofloxacin, a new carboxyquinolone, has potent in vitro bactericidal activity against the major aerobic, gram-negative bacillary pathogens that cause human pyoarthroses . We investigated the in vivo efficacy of ciprofloxacin in a rabbit model of septic arthritis due to Escherichia coli . Animals received either ciprofloxacin (80 mg/kg per day) or gentamicin (5 mg/kg per day) . Ciprofloxacin was rapidly bactericidal in vivo and was significantly more effective in reducing the numbers of E . coli in synovial tissue than was gentamicin at days 10 and 17 of therapy (P less than .0005 and P less than .05, respectively) . Similarly, ciprofloxacin was significantly more active than was gentamicin in reducing the numbers of E . coli in joint fluid on day 10 of therapy (P less than .0005); however, by day 17 of therapy, the numbers of E . coli in joint fluid were not significantly different in the two therapy groups . Neither regimen was effective in preventing the development of postinfectious inflammatory synovitis . There was no in vivo development of resistance to either antibiotic during therapy . Ciprofloxacin therapy was associated with significantly higher bactericidal titers in serum and joint fluid than were observed with gentamicin therapy (P less than .0005) . Ciprofloxacin warrants further in vivo evaluation in invasive E . coli infections.

Eur J Biochem, 1985 Sep 16, 151(3), 613 - 9
Turgor-controlled K+ fluxes and their pathways in Escherichia coli; Meury J et al.; Escherichia coli like most gram-negative bacteria with walls maintains a cytoplasmic osmolarity exceeding that of the medium; the resulting hydrostatic pressure (turgor pressure) pushes the cytoplasmic membrane against the peptidoglycan and creates a tension in the two envelopes . Potassium is the only cation which takes part in the regulation of cellular osmolarity . The adaptation of intracellular K+ concentration to external osmolarity involves K+ turgor-controlled fluxes . When the medium osmolarity is raised an osmodependent influx of K+ can be observed; this is carried out by the K+ transport system TrkA which can also taken up rubidium . A specific and unidirectional pathway allows K+ ions to flow out of the cell when the medium osmolarity is decreased; this pathway reveals two characteristics: it has no affinity for rubidium and it can be blocked by the blockers of eukaryotic K+ channels . Osmodependent fluxes are turned on immediately after the medium osmolarity is disturbed; in contrast, they are turned off gradually as the rate of K+ fluxes approach zero . The rate of K+ influx seems to depend on the level of internal osmolarity and not on the extent of the increase in medium osmolarity . The rate of the efflux is directly proportional to the decrease in medium osmolarity and is independent on the level of internal osmolarity.

Eur J Biochem, 1985 Sep 2, 151(2), 209 - 16
Second lytic target of beta-lactam compounds that have a terminal D-amino acid residue; Tsuruoka T et al.; A new biochemical mechanism of lysing bacterial cells by treatment with certain beta-lactam compounds that possess a terminal D-amino acid moiety in their side chain was demonstrated . The two functions of the molecule, the beta-lactam and terminal D-amino acid moiety, are both involved in the activity of lysing gram-negative bacteria, which is characterized by very rapid lysis of the cells in the first few hours after their contact with the compound . This mechanism was proved by studies on one such compound, named MT-141, which contains a terminal D-cysteine moiety with free amino and carboxyl groups in the 7 beta-side chain of the 7 alpha-methoxy-cephalosporin skeleton . This compound bound to the cell-wall peptidoglycan of Escherichia coli through the D-amino group of its terminal D-amino acid moiety and this seemed to cause rapid cell lysis . Both activities, of binding to peptidoglycan and of causing rapid cell lysis, were inhibited by certain D-amino acids, but not by L-amino acids . Mutants were isolated that had simultaneously gained decreased sensitivity to this kind of beta-lactam compound and supersensitivity to globomycin, an inhibitor of formation of lipoproteins which function in linking the peptidoglycan to the outer membrane . These results suggest that binding of the terminal D-amino acid moiety of the beta-lactam compound to peptidoglycan somehow influences formation of the linkage between the outer membrane and the peptidoglycan and consequently enhances the cell lytic activity of the beta-lactam portion of the molecule.

J Bacteriol, 1985 Sep, 163(3), 1038 - 46
Waveform analysis and structure of flagella and basal complexes from Bdellovibrio bacteriovorus 109J; Thomashow LS et al.; The structure of sheathed flagella from Bdellovibrio bacteriovorus was investigated . The first three periods of these flagella were characterized by progressively smaller wavelengths and amplitudes in periods more distal to the cell . The damped appearance was due to a single nonrandom transition between two helical structures within each filament . The intersection of the two helices, one of which was a threefold-reduced miniature of the other, occurred at a fixed distance along the filament and resulted in a shift in the flagellar axis . Flagella increased in length as the cells aged and assumed a constant miniature waveform at their distal ends . The core filament was the principal determinant of flagellar morphology . It was composed of 28,000- and 29,500-dalton polypeptides . The 28,000-dalton subunits were located in the cell-proximal segment of the filament, and the 29,500-dalton subunits were located in the more distal region . The heteromorphous appearance of bdellovibrio flagella arose from the sequential assembly of these subunits . The basal complex associated with core filaments was examined because of its potential involvement in sheath formation . Bdellovibrio basal organelles were generally similar to those of other gram-negative species, but appeared to lack a disk analogous to the outer membrane-associated L ring which is a normal component of gram-negative basal complexes.

Br J Radiol, 1985 Sep, 58(693), 881 - 4
Anti-lipopolysaccharide toxin therapy for whole body X-irradiation overdose; Gaffin SL et al.; Death in humans from ionising radiation overexposure in the 3-8 Gy (300-800 rad) range is in part due to the toxaemia caused by the entry of gram-negative bacteria and/or their lipopolysaccharide toxin (LPS) into the blood circulation through the walls of partially denuded gut . Anti-LPS hyperimmune equine plasma was evaluated for its ability to lower irradiation-induced lethality . Mice were irradiated with 6.3 Gy (630 rad) and six days later received equine Anti-LPS hyperimmune plasma, control plasma or saline . Mortalities in the three groups were 58%, 92% and 79% (p less than 0.01) respectively . Thus Anti-LPS may prove useful as an adjunct to conventional therapy in treating radiation sickness.

Clin Exp Immunol, 1985 Sep, 61(3), 562 - 8
Factors contributing to the development of cerebral malaria . II . Endotoxin; Usawattanakul W et al.; Limulus amoebocyte lysate test (LALT) was used to detect endotoxin-like substances in the plasma of 15 patients with cerebral malaria, 28 patients with uncomplicated falciparum malaria and 30 healthy controls . On admission, 67% of cerebral malaria patients were positive, whereas only 21.4% of uncomplicated malaria patients and none of controls were positive . Among uncomplicated malaria cases, four of eight patients with parasitaemia over 90,000/mm3 were LALT positive whereas only two of 20 patients with parasitaemia of less than 90,000/mm3 were positive . A follow-up study in cerebral malaria patients showed some variation in LALT positivity rate from day to day (85.7% on day 1, 53.3% on day 3 and all negative on discharge from hospital) . LALT positivity bore no relationship to gram negative bacteraemia . Leucocytosis and elevated serum enzymes were more frequently found in LALT-positive patients . Our results suggest that endotoxin (LALT positivity) of the plasma of malaria patients is derived from either the parasites themselves or from the gut . It relates to parasitaemia, leucocytosis and elevated serum enzymes, but not to the clinical syndrome of cerebral malaria.

J Gen Microbiol, 1985 Sep, 131 ( Pt 9), 2349 - 57
The outer membrane of Treponema pallidum: biological significance and biochemical properties; Penn CW et al.; Rabbits infected intravenously with Treponema pallidum were not markedly febrile, and the pyrogenicity of treponeme preparations administered intravenously to rabbits was negligible . The antibiotic polymyxin B did not induce any ultrastructural changes on the treponemal surface and was not lethal (immobilizing) for T . pallidum, which was, however, highly susceptible to detergents such as SDS . Extraction of treponemes with Triton X-100 removed the outer membrane (despite the presence of Mg2+) as shown by electron microscopy, and solubilized a limited number of proteins detectable by SDS-PAGE, including a dominant antigen (47 kDal) demonstrated by immunoblotting . None of the proteins were heat-modifiable . Periodic acid-silver staining of polyacrylamide gels for carbohydrate together with protease K digestion did not demonstrate major carbohydrate components in whole treponemes, or in the Triton-soluble fraction . Surface iodination of intact treponemes revealed very little surface exposure of treponemal proteins, although a protein which co-migrated with host albumin was labelled and appeared to be associated with the treponemal surface . Many treponemal proteins were, however, labelled when iodination was done in the presence of Triton . These observations, indicate that the outer membrane of T . pallidum differs significantly from those of many Gram-negative pathogens.

J Clin Invest, 1985 Sep, 76(3), 985 - 90
Pretranslational regulation of the synthesis of the third component of complement in human mononuclear phagocytes by the lipid A portion of lipopolysaccharide; Strunk RC et al.; The third component of complement (C3) is a plasma glycoprotein with a variety of biologic functions in the initiation and maintenance of host response to infectious agents . While the hepatocyte is the primary source of plasma C3, mononuclear phagocytes contribute to the regulation of tissue availability of C3 . Lipopolysaccharide (LPS), a constituent of cell walls of gram-negative bacteria, consists of a polysaccharide moiety (core polysaccharide and O antigen) covalently linked to a lipid portion (lipid A) . Using metabolic labeling with {35S}methionine, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis, we examined the effects of LPS on synthesis of C3 by human mononuclear phagocytes as well as synthesis of the second component of complement (C2), factor B, lysozyme, and total protein . LPS increased C3 synthesis 5-30-fold without affecting the kinetics of secretion of C3 or the synthesis of C2, lysozyme, or total protein . Factor B synthesis was consistently increased by LPS . Experiments with lipid A-inactivated LPS (alkaline treated), LPS from a polysaccharide mutant strain, and lipid X (a lipid A precursor) indicated that the lipid A portion is the structural element required for this effect . Northern blot analysis demonstrated at least a fivefold increase in C3 mRNA in LPS-treated monolayers, which suggests that the regulation of the increase in C3 synthesis is pretranslational . C2 mRNA and factor B mRNA were increased approximately twofold . The availability of specific gene products in human mononuclear phagocytes that respond to LPS should permit understanding of the molecular regulation of more complex functions of these cells elicited by LPS in which multiple gene products are coordinately expressed.

Clin Nephrol, 1985 Sep, 24(3), 142 - 6
Effect of hemodialysis on ceftazidime pharmacokinetics; Nikolaidis P et al.; Ceftazidime, a new parenteral third generation cephalosporin, is widely used because of its broad aerobic gram-negative bacterial coverage and its apparent low risk of toxicity . To establish the effect of hemodialysis on the elimination kinetics of ceftazidime and to determine the supplementary dose of the drug after each dialysis session, we studied nine anuric patients with end stage chronic renal failure undergoing hemodialysis . After a single 1 g intravenous bolus injection of ceftazidime a two-compartment open model was used to calculate the pharmacokinetic parameters . The mean ceftazidime concentrations in hemodialysis patients were 64.3 micrograms/ml at the start of dialysis and 20.0 micrograms/ml at the end of dialysis session . A 4-h hemodialysis procedure reduced the area under concentration vs time curve from 795.1 mg/l X h to 175.8/l X h and the elimination half life of ceftazidime from 33.6 h to 3.3 h . Dialyzer clearance was 55.6 ml/min and 55% of the administered dose was recovered in the dialysate fluid . Hemodialysis patients should receive a supplemental dose equal to half their usual maintenance dose immediately after each dialysis session.

Inflammation, 1985 Sep, 9(3), 297 - 308
Endotoxin tolerance diminishes certain antiinflammatory effects of endotoxin; Rosenbaum JT et al.; Endotoxin (bacterial lipopolysaccharide, LPS) is paradoxically both inflammatory and antiinflammatory . A single intravenous injection of 100 micrograms Escherichia coli LPS markedly inhibits the inflammatory changes associated with cutaneous reversed passive Arthus (RPA) reactions in New Zealand white rabbits . Polymorphonuclear (PMN) leukocytes from LPS-treated rabbits exhibit diminished responsiveness in vitro to complement (C5) -derived peptides . Repeated injections of LPS render animals "tolerant", that is, refractory to the toxic and inflammatory effects of LPS . We examined whether tolerance would enhance the ability of LPS to inhibit inflammation not attributable to LPS . Surprisingly, as compared with rabbits receiving a single dose of LPS, tolerant rabbits demonstrated greater inflammatory changes (i.e., PMN exudation, vascular permeability) associated with RPA reactions . PMNs from LPS-tolerant rabbits responded in vitro to C5-derived peptides significantly more than PMNs from rabbits that received a single dose of LPS . We speculate that some antiinflammatory effects of LPS require the toxic or inflammatory effects of LPS itself . These observations might relate to the limited efficacy of fever therapy and the variable effects of gram-negative sepsis on functions of human PMNs.

S Afr Med J, 1985 Aug 31, 68(5), 344 - 5
Aspiration of oral contents in Parkinson's disease . A case report; Cooper RG et al.; A patient with Parkinson's disease developed a non-ketotic hyperosmolar diabetic coma precipitated by chest infection . Initial improvement from treatment with intravenous insulin, ampicillin and fluid therapies was followed by severe deterioration and hypovolaemic shock . Further improvement occurred only when therapy directed against Gram-negative sepsis was added . A barium examination later demonstrated aspiration of oral contents with pulmonary soilage . The differences between the easily recognized early fulminating 'aspiration syndrome' caused by aspiration of gastric contents of low pH and the aspiration of oral contents, which may remain occult for many hours, is highlighted . Life-threatening Gram-negative or anaerobic infection may then occur but remain undiagnosed because the original aspiration of foreign material is unsuspected.

Zh Mikrobiol Epidemiol Immunobiol, 1985 Aug, (8), 87 - 91
{Detection of antibodies neutralizing the endotoxins of gram-negative bacteria using the liposomal potentiometric method}; Vlasov GS et al.; For the first time the study of the indicator system consisting of sensitized liposomes with NaF incorporated as a marker and a fluorine-selective electrode has been made and, as a result, the possibility of the potentiometric determination of the immune lysis of liposomes in the presence of complement and specific antibodies has been demonstrated . The dissolution of the lipid components (Re-chemotype glycolipid and lipid A) in the bilayer matrix obviates the necessity for converting lipid antigens into the water-soluble state in the process of serological tests . As compared with other methods, the liposomal potentiometric method for the determination of Re-chemotype glycolipid and lipid A is highly sensitive (20-40 ng/ml), rapid, technically easy to perform, cheap and does not require large volumes of samples . The disadvantages of this analytical system are the instability of liposomes and the diffusion of fluorine ions from the internal aqueous phase of vesicules . For this reason the immunoassay can be made only within 12 hours after the preparation of sensitized liposomes incorporating the marker.

Appl Environ Microbiol, 1985 Aug, 50(2), 356 - 63
Microbiology and ration digestibility in the hindgut of the ovine; Lewis SM et al.; Contents of the terminal ilea, ceca-proximal colons, and terminal recta were obtained from nine sheep, three of which were fed 100% orchardgrass hay, three of which were fed 60% cracked corn-40% orchardgrass hay, and three of which were fed 80% cracked corn-20% orchardgrass hay . The digestibility of dry matter in the ceca was greatest when the all-hay diet was fed, whereas the percentage of cellulose digestion in the ceca increased with increasing levels of concentrate . For all diets, the total volatile fatty acid concentrations were higher in the ceca than in the other two sites . The cecal pH levels decreased with increased corn levels in the diet . The total microbiol numbers per gram of ileal and cecal contents increased in response to feeding of concentrate; however, across all diets, the ileal counts were 8% or less of the cecal counts . In contrast, the cellulolytic microbial numbers in the ilea were 50% or more of those in the ceca and were highest with the all-hay diet . Both bacterial and end product concentrations in the ceca were equivalent to those occurring in rumina . A total of 16 cellulolytic cultures were isolated and characterized from ileal and cecal contents of animals fed all three diets . Seven gram-negative-rod-shaped organisms were identified as Butyrivibrio fibrisolvens, whose capacity to digest cellulose exceeded that of several rumen strains . The guanine-plus-cytosine content of the DNA of one strain of B . fibrisolvens was 38.8 mol%, compared with the only previously reported value for this species of 41.2 mol%.(ABSTRACT TRUNCATED AT 250 WORDS)

J Clin Microbiol, 1985 Aug, 22(2), 320 - 1
Leptotrichia buccalis bacteremia in neutropenic children; Reig M et al.; Two new cases of Leptotrichia buccalis bacteremia in seriously ill patients were described . The anaerobic, gram-negative microorganism L . buccalis was isolated from blood cultures of two children with severe leukopenia . Anaerobic organisms should be taken into account when a standard protective antibiotic chemotherapy is considered in the immunosuppressed host.

J Gen Microbiol, 1985 Aug, 131 ( Pt 8), 2035 - 9
Immunomodulation by myxospores of Myxococcus xanthus; Ruiz C et al.; Glycerol-induced myxospores of Myxococcus xanthus caused non-specific modulation of humoral and cellular immune responses in laboratory animals . The number of cells which formed specific haemolysins in spleens of mice immunized with sheep erythrocytes was increased when 0.5 X 10(8) myxospores were inoculated 2 d after the erythrocytes, and decreased when myxospores were injected 2 d before or at the same time as the erythrocytes . Both the IgG primary response and the secondary response to erythrocytes were decreased in rabbits after pretreatment with 2 X 10(8) myxospores per rabbit . Delayed-type hypersensitivity to sheep erythrocytes was also suppressed in mice after intraperitoneal (i.p.) injection of 0.3 X 10(8) myxospores . One day after i.p . injection of myxospores, neither an inflammatory response nor bone marrow cell depletion was observed in mice . These results support the idea that M . xanthus myxospores possess diverse immunomodulation properties apparently due to factors different from the classical LPS of Gram-negative bacteria.

Antimicrob Agents Chemother, 1985 Aug, 28(2), 282 - 8
Comparison of aminoglycoside resistance patterns in Japan, Formosa, and Korea, Chile, and the United States; Shimizu K et al.; The resistance mechanisms of more than 2,000 aminoglycoside-resistant gram-negative aerobic bacteria were estimated by a method that assigned a biochemical mechanism based on susceptibility to selected aminoglycosides . Strains from hospitals in Japan, Formosa, and Korea (the Far East) were compared with strains from Chile and the United States . Of the strains from Chile, 90% had an aminoglycoside resistance pattern indicative of the 3-N-acetyltransferase {AAC(3)-V} enzyme . Of the strains from the Far East, 78% had susceptibility patterns suggesting the presence of AAC(6') enzymes . In contrast, strains from the United States had a wider variety of resistance mechanisms including 2''-O-adenylyltidyltransferase {ANT(2'')}, AAC(3), AAC(6'), and AAC(2') . Reflecting these differences in resistance patterns, the frequencies of resistance to gentamicin, tobramycin, dibekacin, and amikacin in strains from the United States were different from those in strains from the Far East . These differences seem to be correlated with different aminoglycoside usage in the two regions . In the United States, where gentamicin was the most widely used aminoglycoside, 92% of the strains were resistant to gentamicin, 81% were resistant to dibekacin, and 8.8% were resistant to amikacin . In the Far East, dibekacin and kanamycin were widely used in the past and more recently amikacin has been frequently used . Of the strains from this region, 99% were resistant to dibekacin, 85% were resistant to gentamicin, and 35% were resistant to amikacin.

J Clin Periodontol, 1985 Aug, 12(7), 507 - 24
Ultrastructure of developing subgingival plaque in beagle dogs; ten Napel JH et al.; The aim of this study was to describe the ultrastructure of developing subgingival plaque . In 6 beagle dogs, healthy gingiva prevailed after a pre-experimental period of intensive plaque control . At the start of the experiment, all oral hygiene measures were withdrawn . Biopsies comprising buccal gingiva and adjacent dental tissues were obtained from premolars and molars on days 0, 4, 7, 21, and 28 and processed for electron microscopy . Sections of day 0 exhibited a junctional epithelium in close contact with the tooth and absence of subgingival aggregations of bacteria on the tooth surface . For each of the subsequent periods studied, some sections did not exhibit subgingival plaque . In the remaining sections with subgingival plaque, almost all bacteria appeared in aggregates attached to the tooth surface . The number of bacteria at the orifice of the gingival sulcus increased with time . Further apically, the micro-organisms formed either a continuous layer of even thickness or, more frequently, discrete microcolonies along the tooth surface . The bacteria comprised practically entirely Gram-negative cocci, rods or spirochetes . The latter were present either mixed with the other types in the discrete microcolonies or constituted the bulk of the continuous layer of plaque . In the apical part of the subgingival plaque, a shift from Gram-negative cocci and rod during the early stages of gingival inflammation, to a predominance of spirochetes in later stages was noted . The study shows that in dogs, starting from a plaque- and gingivitis-free baseline, a predominantly Gram-negative subgingival flora may form within a few days after withdrawal of all oral hygiene measures and that a shift in the bacterial composition of the subgingival plaque takes place during a 4-week period of no oral hygiene.

Appl Environ Microbiol, 1985 Aug, 50(2), 441 - 6
Recombinant plasmid conferring proline overproduction and osmotic tolerance; Jakowec MW et al.; A recombinant plasmid carrying the proBA (pro-74) mutant allele which governs osmotic tolerance and proline overproduction was constructed by using the broad-host-range plasmid vector pQSR49 . The physiological, biochemical, and genetic properties of strains carrying the pQSR49 derivatives pMJ101 and pMJ1, mutant and wild type, respectively, were investigated . pMJ101 conferred enhanced osmotolerance compared with strains carrying the wild type, pMJ1 . These results are in contrast to those obtained previously with strains carrying recombinant plasmids based on pBR322 that failed to confer the osmotic tolerance phenotype . gamma-Glutamyl kinase (first step in proline biosynthesis) from strains carrying pMJ101 was 200-fold less sensitive to feedback inhibition than was the wild-type enzyme . As expected, the intracellular proline levels of strains carrying pMJ101 were more than an order of magnitude higher than those of the wild type . An analysis of copy number revealed that the pQSR49 constructs were present in the cell at a level six- to eightfold lower than those of the pBR322 recombinants, which may account for the difference in phenotype . We found that the genetic stability of the pQSR49 derivative in a variety of gram-negative bacteria was dependent on the insert orientation and the presence of foreign DNA on the plasmid . These factors may be significant in future studies aimed at expanding the osmotolerance phenotype to a broad range of gram-negative bacteria.

Am J Med, 1985 Jul 15, 79(1A), 28 - 36
Role of amikacin in the management of intra-abdominal sepsis; Dougherty SH; Controversy has developed regarding the antibiotic management of intra-abdominal sepsis because of the recent availability of the third-generation cephalosporins and ureidopenicillins as alternatives to traditional combination therapy (aminoglycosides plus clindamycin) . Most observers now acknowledge the need to provide anti-anaerobic as well as anti-aerobic gram-negative drug coverage . Although most of the newer agents do provide such broad-spectrum coverage, doubt remains regarding their efficacy because of flaws in comparative study design and the observation that resistance to the newer agents, which may even extend to the aminoglycosides, can emerge in individual patients during single courses of antibiotic therapy . Indeed, such resistance is most likely to occur during the treatment of seriously ill, immunodepressed patients who have undergone multiple reoperation for persistent or recurrent intra-abdominal sepsis--the precise group for which the new drugs were most desired as less toxic alternatives to the aminoglycosides . On the basis of such observations, combination therapy with the aminoglycosides, appears to remain the most logical choice . In the setting of nosocomial sepsis and pathogen resistance to other aminoglycosides, amikacin may be especially effective . Recent surveillance data indicate that the use of amikacin under such circumstances not only may provide effective antibiotic therapy, but also may actually reduce the level of microbial resistance to the other aminoglycosides . Past concern regarding the development of resistance to amikacin has probably been excessive and should not deter the use of this agent under appropriate clinical circumstances.

Am J Med, 1985 Jul 15, 79(1A), 37 - 42
Three-year experience with amikacin sulfate as an exclusive surgical aminoglycoside in a large acute-care hospital; Lee JT Jr; Aminoglycosides have important roles as perioperative adjunctive antibiotics in the surgical management of peritonitis . In the past, most surgeons have used gentamicin in combination with a drug aimed at intraperitoneal anaerobic pathogens, either clindamycin or metronidazole . Amikacin has been traditionally reserved for culture-proved infections due to gram-negative organisms resistant to gentamicin or other aminoglycosides . At the Minneapolis Veterans Administration Medical Center, a worrisome incidence of gentamicin-resistant hospital isolates led to a decision to make amikacin the exclusive, routine surgical aminoglycoside for all abdominal infections, as well as all hospital-acquired infections in surgical patients involving gram-negative aerobes and requiring parenteral therapy . This report describes the resultant three-year experience with amikacin in surgical patients . Data from four research studies involving these patients suggest that exclusive amikacin use has not led to the emergence of amikacin-resistant organisms or to significant nephrotoxicity . Amikacin use in surgical patients is supported in hospital environments where gentamicin resistance is judged to be a significant clinical risk factor.

Lancet, 1985 Jul 13, 2(8446), 59 - 63
Prevention of gram-negative shock and death in surgical patients by antibody to endotoxin core glycolipid; Baumgartner JD et al.; The prophylactic effect of antibody to endotoxin core glycolipid was studied in surgical patients at high risk of gram-negative infection . At randomisation (on admission to intensive care unit), every 5 days thereafter, and at onset of septic shock, patients received plasma taken from donors before (control) or after immunisation with Escherichia coli J5, a mutant with only core determinants in its endotoxin . Gram-negative shock occurred in 15 of 136 controls and 6 of 126 J5 antibody recipients and related deaths in 9 of 136 and 2 of 126, respectively . J5 antibody was most effective in abdominal surgery patients, in whom shock occurred in 13 of 83 controls and 2 of 71 antibody recipients . Although antibody prophylaxis did not lower the infection rate, it prevented the serious consequences of gram-negative infections and thus improved the overall prognosis.

J Infect Dis, 1985 Jul, 152(1), 177 - 84
Effects of specific antibodies, hormones, and lipoproteins on bacterial lipopolysaccharides injected into the rat; Munford RS et al.; When gram-negative lipopolysaccharides (LPS) are injected intravenously into experimental animals, approximately one-third of the LPS bind to high-density lipoproteins (HDL) in the plasma and are slowly taken up into tissues by HDL-mediated mechanisms . Molecules of LPS in the plasma that do not bind to HDL are taken up more rapidly by tissues that are rich in phagocytic cells (e.g., liver, spleen) . In these experiments we evaluated the effects of several potential host factors on the binding of LPS to HDL and on the tissue uptake of LPS injected intravenously into rats . Antibodies to LPS inhibited LPS binding to HDL and increased the uptake of the injected LPS (as well as the LPS in preformed LPS-HDL complexes) by the liver and spleen . High levels of circulating HDL decreased the uptake of injected LPS and LPS-HDL complexes by the adrenal gland, presumably by occupying tissue receptors for HDL . Pretreating rats with dexamethasone unexpectedly decreased the uptake of injected LPS-HDL complexes by the adrenal gland and increased uptake of LPS by the gland, a result suggesting that this drug has opposing effects on the uptake of HDL-bound and -unbound LPS by the adrenal gland . The host factors studied appear to influence the fate of injected LPS by acting at different sites: IgG antibody to LPS blocks the binding of LPS to HDL in the plasma, whereas all of the factors studied have effects that modulate the uptake of LPS or LPS-HDL complexes or both by the cells of specific tissues.

Arch Intern Med, 1985 Jul, 145(7), 1212 - 6
Improved mortality in gram-negative bacillary bacteremia; McCue JD; From 1979 to 1982, the four years of this study, episodes of gram-negative bacillary bacteremia occurred in a 489-bed community teaching hospital--an increase of 15.9% . Mortality related to bacteremia was 19.4% overall and only 3.2% for the 158 episodes involving nonfatal underlying illnesses, lower figures than those reported in the past . The severity of underlying illnesses in bacteremic patients dominated all other clinical variables that were studied as prognostic factors for the outcome of the episode . The same bacteremia-related mortality was seen in patients who had empirically received (1) multiple-antibiotic regimens in which one or more drugs were active against the pathogenic organism(s), (2) either an appropriate aminoglycoside or beta-lactam antibiotic alone, or (3) both an aminoglycoside antibiotic and a beta-lactam antibiotic active against the pathogenic organism(s).

Vestn Khir Im I I Grek, 1985 Jul, 135(7), 35 - 7
{Antibiotic therapy in the surgical treatment of suppurative cholangitis}; Zemskov VS et al.; The pathogenic agents of purulent cholangitis are known to be the conditionally pathogenic mainly gram-negative bacteria of the intestinal group . Kanamycin, monomycin, furazolidon should be used . The antibiotic treatment should be corrected according to data of inoculations of blood taken during the first days of staying in the hospital, then to data of inoculations of bile taken both during operation and at the 3d-4th postoperative days.

J Assoc Off Anal Chem, 1985 Jul-Aug, 68(4), 661 - 4
Sensitive enzyme immunoassay of colistin and its application to detect residual colistin in rainbow trout tissue; Kitagawa T et al.; An antibody against colistin (CL), an antibiotic effective for gram-negative bacteria, was produced in rabbits immunized with a colistin-protein conjugate . The conjugate was prepared by a novel and convenient procedure devised to couple an amino group of CL to thiol groups of bovine serum albumin (BSA) introduced by thiol exchange reduction of its disulfide bonds with dithiothreitol, using N-(m-maleimidobenzoyloxy)succinimide (MBS) as a cross-linker . Enzyme labeling of CL with beta-D-galactosidase was performed by utilizing another cross-linker, N-(gamma-maleimidobutyryloxy)succinimide, by means of a convenient labeling method . A double antibody enzyme immunoassay of CL, which could determine as little as 30 ng/mL of CL, was developed using labeled CL and anti-CL antiserum . With this assay, drug levels were easily determined in fish tissue after CL administration . The enzyme immunoassay should provide a useful tool for detection and quantitation of residual drugs in foods and related products.

Antibiot Med Biotekhnol, 1985 Jul, 30(7), 495 - 8
{Isoelectric focusing of different classes of beta-lactamases in cell-free extracts of Gram-negative bacteria}; Sazykin AIu; A method of electrofocusing of beta-lactamases in cell-free bacterial extracts was developed . It provided determination of the isoelectric points of beta-lactamases without preliminary preparation of the homogeneous enzyme . Isoelectrofocusing of beta-lactamases of 14 poly-resistant strains of gram-negative bacteria of six species was performed . It was found that the isoelectric points of the beta-lactamase belonging to the same class according to Richmond were different . No species specificity of the beta-lactamases of the gram-negative bacteria was observed.

G Batteriol Virol Immunol, 1985 Jul-Dec, 78(7-12), 144 - 54
{Generation of pro-coagulation activity by leukocytes stimulated in vitro with lipoglycans from mycoplasmas}; Monno RA et al.; Lipoglycans represent a special type of lipopoly-saccharide that differs in structure from the well-known gram-negative bacteria lipopolysaccharide . After briefly describing their most important characteristics, the authors take into consideration the in vitro interaction between lipoglycans from Acholeplasma granularum, Acholeplasma oculi, and Acholeplasma axanthum and human leukocytes in terms of production of procoagulant activity . The results obtained show that the examined lipoglycans possess, similarly to lipopolysaccharides, the capacity to induce the production of procoagulant activity, thromboplastin-like, from human mononuclear cells . However, the pathophysiological significance of this endotoxin-like activity remains to be established.

Arch Microbiol, 1985 Jun, 142(1), 6 - 11
Nature and linkage type of fatty acids present in lipopolysaccharides of phase I and II Coxiella burnetii; Wollenweber HW et al.; The constituent fatty acids of lipopolysaccharides (LPS) of Coxiella burnetii (phase I and II) were qualitatively and quantitatively analysed by combined gas-liquid chromatography/mass spectrometry . The total fatty acid content (per mg LPS) was determined as 90.0 nmol (2.3 wt%) for LPS of phase I cells (LPS I) and 179.1 nmol (4.8 wt%) for LPS of phase II cells (LPS II) . Of the 24 different acyl residues characterized (12 to 18 carbon atoms), nine were 3-hydroxy fatty acids (normal, iso- and anteiso-branched) which quantitatively predominated . All 3-hydroxylated fatty acids were found to possess the (R)-configuration, to be exclusively amide-linked and to be acylated at their 3-hydroxyl group . Ester-linked nonhydroxylated fatty acids (normal, iso- and anteiso-branched) were present but ester-bound 3-hydroxy- or 3-acyloxyacyl residues were lacking from C . burnetii LPS I and LPS II . As the major acyl group (R)-3-(12-methyl-tetradecanoyloxy)-12-methyl-tetradecanoic acid was identified . Our results show that the complex fatty acid spectrum of C . burnetii differs considerably from that of LPS of other Gram-negative bacteria . They further suggest an enormous heterogeneity of the lipid A component of C . burnetii LPS I and LPS II.

Scand J Work Environ Health, 1985 Jun, 11(3 Spec No), 199 - 206
Organic dusts and lung reactions--exposure characteristics and mechanisms for disease; Rylander R; Exposure to organic dusts has been related to pulmonary occupational diseases in a variety of environments . These dusts contain several different agents, but the microbial contamination is always important, particularly regarding molds and Gram-negative bacteria . In the lung, organic dusts cause a series of reactions ranging from irritation with neutrophilic invasion to the initiation of cell mediator release and the development of antibodies . For certain diseases, such as byssinosis, the relation between the cell reactions and the disease is relatively well established; for others, such as allergic alveolitis (hypersensitivity pneumonitis) with fibrosis, more research is needed before the pathogenesis can be established . An understanding of the specific agents causing the disease and the cellular reactions behind its development is essential for its prevention.

J Clin Microbiol, 1985 Jun, 21(6), 865 - 8
Detection of endotoxin in the plasma of patients with gram-negative bacterial sepsis by the Limulus amoebocyte lysate assay; Pearson FC et al.; A total of 120 Limulus amoebocyte lysate (LAL) determinations were made on plasma obtained from normal, healthy human blood donors . Results demonstrated a mean endotoxin level in blood of 0.02 to 1.57 pg/ml . The amount of Escherichia coli endotoxin added to human plasma samples can be quantitated by both nephelometry and turbidimetry . Endotoxin-spiked samples were shown to be significantly different from unspiked samples . When plasma samples were collected from 45 patients hospitalized at three centers, a strong association was demonstrated between a positive Limulus amoebocyte lysate assay and a septic condition . Sensitivity, specificity, and false-positive and false-negative rates for the Limulus amoebocyte lysate assay as a diagnostic test for gram-negative bacteremia were estimated.

Southeast Asian J Trop Med Public Health, 1985 Jun, 16(2), 291 - 5
Pure red cell aplasia in Thailand: report of twenty four cases; Jootar S et al.; Twenty four cases of pure red cell aplasia were reported . No underlying diseases were found in two cases . Of the 22 cases with secondary form, 10 were from infections, mostly gram negative organisms . Three cases had systemic lupus erythematosus, two had autoimmune hemolytic anemia . The following conditions were found in one each: thymoma, thyroid carcinoma, protein calorie malnutrition, rheumatoid arthritis, non-Hodgkin lymphoma and Sheehan's syndrome . Three patients died, two from uncontrolled infection, the other from uncontrolled SLE and subsequently systemic fungal infection . Only one of the 2 primary cases responded to immunosuppressive drugs . The majority of patients with underlying infections, PRCA resolved after the infections were treated . This is the first reported series of PRCA in Thailand.

J Clin Microbiol, 1985 Jun, 21(6), 955 - 8
Evaluation of a lysis direct plating method for pediatric blood cultures; Welch DF et al.; The Isolator 1.5 Microbial tube (E . I . du Pont de Nemours & Co., Inc., Wilmington, Del.) is a commercially available blood culture system for use in pediatrics . The methodology is based on blood lysis followed by direct plating of the sample on culture media to detect bacteria and fungi . Comparative recovery rates of pathogens from blood collected in this and a conventional broth system were similar . The Isolator detected 104 of 120 clinically significant isolates, whereas 106 of 120 isolates were detected by the broth system . The major advantage of the Isolator methodology was early detection of septicemia . Initial detection of gram-negative bacteria occurred an average of 14.2 h earlier by the Isolator system than by the conventional broth method . The Isolator also permitted quantitation of bacteremia and fungemia . Probable contaminants were recovered from 10.0% of the cultures processed by the Isolator, but steps which could be taken to minimize this problem were identified . The Isolator is a useful method for pediatric blood cultures.

Am J Physiol, 1985 Jun, 248(6 Pt 1), E732 - 40
Effect of sepsis on VLDL kinetics: responses in basal state and during glucose infusion; Wolfe RR et al.; The effect of gram-negative sepsis on the kinetics and oxidation of very low-density lipoprotein (VLDL) fatty acids was assessed in conscious dogs in the normal state and 24 h after infusion of live Escherichia coli . VLDL, labeled with {2-3H}glycerol and {1-14C}palmitic acid, was used to trace VLDL kinetics and oxidation, and {1-13C}palmitic acid bound to albumin was infused simultaneously to quantify kinetics and oxidation of free fatty acid (FFA) in plasma . Sepsis caused a fivefold increase in the rate of VLDL production (RaVLDL) . In the control dogs, the direct oxidation of VLDL-fatty acids was not an important contributor to their overall energy metabolism, but in dogs with sepsis, 17% of the total rate of CO2 production could be accounted for by VLDL-fatty acid oxidation . When glucose was infused into dogs with insulin and glucagon levels clamped at basal levels (by means of infusion of somatostatin and replacement of the hormones), RaVLDL increased significantly in the control dogs, but it did not increase further in dogs with sepsis . We conclude that the increase in triglyceride concentration in fasting dogs with gram-negative sepsis is the result of an increase in VLDL production and that the fatty acids in VLDL can serve as an important source of energy in sepsis.

J Infect Dis, 1985 Jun, 151(6), 988 - 94
Age-related prevalence of human serum IgG and IgM antibody to the core glycolipid of Escherichia coli strain J5, as measured by ELISA; Law BJ et al.; Administration of antibody to Escherichia coli strain J5 reduces mortality in patients with gram-negative septic shock . Limited evidence suggests that this antibody is present in the serum of healthy individuals and deficient in persons with malignancy; however, the age-related prevalence of serum antibody to strain J5 in normal subjects is unknown . We developed an ELISA method to measure IgG and IgM antibody to purified J5 lipopolysaccharide (LPS) in sera from 33 maternal-newborn cord pairs, 40 neonates, and 253 individuals aged from one day to twenty years . Reciprocal geometric mean titers for serum antibody to strain J5 LPS were as follows: maternal-IgG, 12.3; IgM, 174; one month old or less-IgG, 8.3; IgM, 5.6; one to 24 months old-IgG, 10.5; IgM, 44.7; greater than 24 months old -IgG, 16.2; IgM, 200 . We conclude that human serum antibody to J5 LPS is predominantly IgM and that it is usually present by two years of age . The ELISA method should provide a valuable tool in studying the relation of core glycolipid antibody to immunity and therapy of gram-negative bacterial infections.

J Infect Dis, 1985 Jun, 151(6), 1012 - 8
Role of antibiotic class in the rate of liberation of endotoxin during therapy for experimental gram-negative bacterial sepsis; Shenep JL et al.; To evaluate the role of antibiotic class in the rate of liberation of endotoxin during therapy for sepsis caused by gram-negative bacteria, we obtained serial blood samples from rabbits with sepsis caused by Escherichia coli and treated with chloramphenicol, gentamicin, or moxalactam . The concentrations of viable bacteria, free endotoxin, and total endotoxin in each blood sample were measured . In rabbits treated with chloramphenicol, the geometric mean levels of free endotoxin remained proportional to the geometric mean levels of bacteremia, a result indicating the absence of antibiotic-induced endotoxin liberation . In contrast, levels of free endotoxin increased rapidly while levels of bacteremia declined after treatment with gentamicin or moxalactam, a result indicating antibiotic-induced release of endotoxin . Despite similar rates of bacterial killing, mean levels of free endotoxin were as much as 20-fold higher in rabbits treated with moxalactam than in paired rabbits receiving gentamicin (P less than .05) . These results indicate that endotoxin liberation during therapy for sepsis caused by gram-negative bacteria is dependent upon the class of antibiotic administered and is not necessarily correlated with the rate of bacterial killing.

Jpn J Antibiot, 1985 Jun, 38(6), 1648 - 53
{Pharmacokinetic study of cefmenoxime concerning its transfer into tissues}; Washida H et al.; Cefmenoxime (1 g) was intravenously administered by one shot to investigate its pharmacokinetic profile with respect to transfer into the kidney, vesical wall and prostatic adenoma . The concentration of cefmenoxime in the kidney reached a peak of 403 micrograms/g at 0.17 hour after administration . The biological half-life was 0.74 hour . In the vesical wall, the level of cefmenoxime reached a peak of 28 micrograms/g at 0.67 hour after administration . The half-life was 2.30 hours . The peak level of cefmenoxime in the prostatic adenoma was 22 micrograms/g at 0.39 hour after administration . The half-life was 2.90 hours . The concentrations of cefmenoxime in these urogenital tissues were higher than its MIC80 and MBC80 against various Gram-negative organisms.

Infect Control, 1985 Jun, 6(6), 231 - 2
Bedside resuscitation bags: a source of bacterial contamination; Thompson AC et al.; Resuscitation bags at the bedside of 12 intubated ICU patients with known culture positive sputum were evaluated as possible reservoirs of pathogenic bacteria . As oxygen was constantly flowing into the bags, sampling of gas continuously emitted from the exhalation valve assembly was sampled to detect if a bacteria-laden aerosol was being created . Positive cultures were obtained from 75% of the valves and 25% of the aerosol samples . In those patients with gram-negative bacteria in their sputum, 71% of the valves and 29% of the gas samples were positive for the same organisms . Various methods to control this possible contributing factor to nosocomial infection in the ICU are discussed.

J Infect Dis, 1985 Jun, 151(6), 1005 - 11
Failure of monoclonal antibodies to core glycolipid to bind intact smooth strains of Escherichia coli; Gigliotti F et al.; To study the ability of antibody to the core glycolipid (CGL) region of lipopolysaccharide (LPS) to bind intact gram-negative bacterial cells, we produced monoclonal antibodies that bind LPS from the Escherichia coli rough mutant J5 . Four representative monoclonal antibodies that bound four distinct epitopes on the CGL region of LPS were studied . All four antibodies bound both isolated J5 LPS and intact J5 bacterial cells, but none of the antibodies bound to intact cells of E . coli O111:B4 or K1:O7 . Binding of the monoclonal antibodies to isolated LPS from these latter two smooth strains was variable . These results confirm the presence of shared antigenic sites in the CGL region of heterologous LPS molecules but indicate that these sites are not necessarily available on smooth gram-negative bacteria for binding by antibody to CGL.

Infection, 1985 May-Jun, 13(3), 120 - 4
Preliminary study on administration of high-titer lipid A antibody serum in sepsis and septic shock patients; Marget W et al.; To determine whether sera containing high titers of lipid A antibody may be applicable at effective doses in treating patients with septicemia, a pharmacokinetic study was performed in seven patients aged 24 to 73 years with gram-negative septicemia and various underlying diseases, one of whom received a placebo, and in one patient without infection . In this study, we attempted to determine the effective dosage, the number of infusions and appropriate administration interval for the prevention and treatment of endotoxin shock . Blood samples from each patient were tested for lipid A antibodies before and at regular intervals after administration using an enzyme-linked immunosorbent assay (ELISA) . Selected, pooled human immunoglobulin preparations containing high titers (expressed as exponents of 10) of IgG (titer = 3-4) and IgM (titer = 2-4) lipid A antibodies were administered first at a dosage of 8 ml/kg body weight . After administration, the mean lipid A antibody titer increased from 0.4 to 2.3 for IgG and from 2 to 2.4 for IgM . The initial increase was followed by a drop in titer within 24 h, which was perhaps due to antibody consumption . Following the second administration (24 h after the first) of only 4 ml/kg body weight, the mean IgG and IgM titers increased to 2.4 and 3.3, respectively, and dropped slower . A notable increase in circulating lipid A antibody titers was achieved, and four of six treated patients recovered from the sepsis . The two patients who died entered the study in a pre-terminal state . These studies encouraged us to initiate a randomized, double-blind controlled study.

Infection, 1985 May-Jun, 13(3), 115 - 9
Serum antibodies against common antigens of bacterial lipopolysaccharides in healthy adults and in patients with multiple myeloma; Stoll C et al.; The incidence of infections caused by gram-negative bacteria is increased in patients with multiple myeloma due to secondary humoral immunodeficiency . In order to diagnose patients with increased susceptibility to gram-negative infections, serum antibodies against common determinants of lipopolysaccharides (lipid A and core-polysaccharide) were determined by a rapid enzyme-linked immunosorbent assay (ELISA) . It was possible to define a group of patients at high risk of contracting gram-negative infections using this test . Intravenous IgG preparations used as a substitute were shown to contain antibodies against these common antigens . However, it is suggested that the clinically recognized efficacy of these preparations could be due to their containing anti-LPS antibodies.

Antimicrob Agents Chemother, 1985 May, 27(5), 858 - 62
Effect of temocillin and moxalactam on platelet responsiveness and bleeding time in normal volunteers; Nunn B et al.; The effects of temocillin and moxalactam on platelet responsiveness and bleeding time were examined in healthy male volunteers . In the first study, moxalactam (4 g intravenously every 12 h) was given to six subjects; template bleeding times were at least doubled in five subjects 12 to 14 h after 7 doses (P = 0.008) and in all six subjects 12 to 14 h after 13 doses (P = 0.004) . ADP-induced primary aggregation was approximately halved after 7 (P = 0.026) and 13 doses (P = 0.008), and there was a markedly increased tendency toward disaggregation . Collagen-induced aggregation was also halved, but the effect only reached statistical significance after 13 doses (P = 0.008) . There was essentially no effect on primary aggregation in response to the thromboxane receptor agonist U46619 or to platelet activating factor . Temocillin (4 g intravenously every 12 h) was given to eight subjects, three of whom had participated in the moxalactam study 8 weeks earlier . Temocillin had no significant effect on template bleeding time 12 to 14 h after 7 or 13 doses . However, in four subjects, the endpoint may have been less abrupt . There was no significant effect on ADP-induced primary aggregation or responsiveness to collagen . Even after 13 doses of temocillin, secondary aggregation in response to normal concentrations of ADP was demonstrable in the platelet-rich plasma of all eight subjects . Neither antibiotic had any effect on prothrombin times . Thus, with methodology that readily detected the effects of moxalactam on hemostasis, we were unable to demonstrate any unequivocal deleterious effects of temocillin at its maximum recommended dose . Temocillin may therefore be particularly useful for the treatment of many gram-negative infections in patients at increased risk of clinical bleeding.

J Bacteriol, 1985 May, 162(2), 693 - 7
Enzymatic release of halogens or methanol from some substituted protocatechuic acids; Kersten PJ et al.; Four strains of gram-negative bacteria capable of growing at the expense of 5-chlorovanillate were isolated from soil, and the metabolism of one strain was studied in particular detail . In the presence of alpha, alpha'-bipyridyl, a suspension of 5-chlorovanillate-grown cells accumulated 5-chloroprotocatechuate from 5-chlorovanillate; in the absence of inhibitor these compounds, and various other 5-substituted protocatechuates and vanillates, were oxidized to completion . Cell suspensions of this strain grown on 5-chlorovanillate or vanillate released chloride quantitatively from 5-chlorovanillate and released methanol from syringate . Extracts of cells grown with 4-hydroxybenzoate, vanillate, or syringate possessed high levels of both protocatechuate 4,5-dioxygenase and 2-pyrone-4,6-dicarboxylate hydrolase; extracts from acetate-grown cells did not . Protocatechuate 4,5-dioxygenase, purified from strains that could grow with 5-chlorovanillate, oxidized 5-halogeno-protocatechuates and 3-O-methylgallate with the formation of 2-pyrone-4,6-dicarboxylate . A crude extract converted 5-chloroprotocatechuate into pyruvate plus oxaloacetate . On the basis of these observations, a meta-fission reaction sequence is proposed for the bacterial degradation of vanillate and protocatechuate substituted at C-5 of the benzene ring with halogen or methoxyl.

Jpn J Antibiot, 1985 May, 38(5), 1319 - 30
{Basic and clinical studies of cefminox in obstetrics and gynecology}; Kubota K; The study was done to evaluate the usefulness of cefminox (CMNX, MT-141) injection for the treatment of infections in the field of obstetrics and gynecology . Fundamental and clinical studies were made and following results were obtained . When 1 g of CMNX was administered by intravenous single shot, the maximum concentrations in various tissues of female genital organs were as follows: 34.98 micrograms/g in oviduct at 1 hour 10 minutes after the single shot, and 37.11, 28.01, 26.84, 30.01 and 40.06 micrograms/g in ovary, endometrium, myometrium, cervix uteri and portio, respectively, after 1 hour 20 minutes . In the clinical studies, CMNX was given to 13 cases with female genital organ infections and others . As for the clinical effects, responses were excellent in 1 case, good in 12 cases among 13 cases in total . The efficacy rate was 100% . As for the clinical effects on causative bacteria, the efficacy rates were 100% (3/3) for single infections due to Gram-negative bacteria, 100% (8/8) for mixed infection cases . Side effects were not observed . CMNX showed a satisfactory clinical efficacy and a potent bacteriological effect in treatment of the infection in the field of obstetrics and gynecology, and it has been concluded that CMNX will be a useful addition to the antibiotics for the therapy of these infections.

Pediatr Infect Dis, 1985 May-Jun, 4(3), 265 - 9
Use of trimethoprim-sulfamethoxazole to prevent bacterial infections in children with acute lymphoblastic leukemia; Goorin AM et al.; We assessed the efficacy of prophylactic antibiotics in children receiving intensive chemotherapy for acute lymphoblastic leukemia . The patients were randomized to receive either trimethoprim-sulfamethoxazole (TMP-SMX) or placebo in a double-blind trial . Thirty patients were evaluated in each group . Children receiving TMP-SMX had fewer episodes of bacteremia (0 vs . 5) and otitis media (3 vs . 18) . The geometric mean of the neutrophil nadir was 172 in the TMP-SMX group and 287 in controls . However, no increased delay or dose reduction of chemotherapy was observed in the TMP-SMX treated patients . Five patients who received TMP-SMX developed Gram-negative rods resistant to TMP-SMX on surveillance stool cultures . We conclude that TMP-SMX prophylaxis decreased certain bacterial infections in children with acute lymphoblastic leukemia without causing clinically significant toxicity . The emergence of Gram-negative rods resistant to TMP-SMX in treated patients suggests that TMP-SMX prophylaxis should be restricted to patients who are at high risk for developing a bacterial infection or Pneumocystis carinii pneumonia.

Infect Immun, 1985 May, 48(2), 464 - 73
Uptake and deacylation of bacterial lipopolysaccharides by macrophages from normal and endotoxin-hyporesponsive mice; Munford RS et al.; Macrophages are thought to play a central role in the responses of animals to gram-negative bacterial lipopolysaccharides (LPS) . Since nothing is known about the metabolism of LPS by these cells, we studied the uptake and deacylation of radiolabeled LPS by thioglycolate-elicited peritoneal macrophages from normal (C3H/HeN) and LPS-hyporesponsive (C3H/HeJ) mice . Macrophages from both kinds of mice took up and deacylated LPS that were added to the culture medium . Opsonization of the LPS with anti-LPS immunoglobulin G antibodies greatly increased LPS uptake; the opsonized LPS also underwent deacylation at rates that were directly related to the amount of cell-associated LPS . An analysis of the fatty acid composition of the cell-associated LPS indicated that the cells have one or more acyloxyacyl hydrolases that remove the non-hydroxylated fatty acids that are normally substituted to the hydroxyl groups of (glucosamine-linked) 3-hydroxytetradecanoate residues in lipid A; we also found evidence for deacylation of 3-hydroxytetradecanoate from the glucosamine backbone . LPS deacylation by macrophages from C3H/HeN and C3H/HeJ mice was qualitatively and quantitatively similar . Nonopsonized LPS are able to stimulate LPS-responsive cells; in these studies we established that animal cells can deacylate nonopsonized LPS, thus raising the possibility that LPS metabolism may play a role in modulating cellular stimulation.

J Am Vet Med Assoc, 1985 Apr 15, 186(8), 798 - 801
Prevalence and causes of ram epididymitis in Utah; Bagley CV et al.; Palpation of the testicles and epididymides was used to determine the prevalence of ram epididymitis (RE) in Utah range flocks over a 5-year period . Of 62 range flocks examined, 58 had rams with palpable lesions . A comparison was made of prevalence by right vs left testicle involvement, but no significant difference was found . Culling all rams with palpable lesions from the flock progressively reduced the annual incidence, but did not eliminate RE from any flock . Semen and specimens of reproductive tract tissue were obtained from representative flocks for bacterial culture . Brucella ovis was found commonly in range flocks, but was not isolated from RE-affected rams in virgin, ram-producing flocks . In the ram-producing flocks, a gram-negative, pleomorphic rod-type organism was found . It was concluded that RE should be considered as 2 disease entities, based on the finding of 2 causative organisms and the management systems under which those organisms are found . Brucella ovis was the major cause in persistently infected range flocks, whereas the gram-negative pleomorphic rod-type organism is the apparent cause of RE in virgin, ram-producing flocks.

Am J Med, 1985 Apr, 78(4), 687 - 90
Cutaneous infection at dog bite wounds associated with fulminant DF-2 septicemia; Kalb R et al.; Severe DF-2 sepsis with disseminated intravascular coagulopathy developed following dog bites in two patients who had undergone prior splenectomy . Eschariform lesions developed at the site of the animal bite in both patients . DF-2 is an unusual animal-borne slow-growing gram-negative rod that can cause fulminant sepsis in splenectomized patients . Splenectomized patients should be aware of the hazards from a dog bite . The presence of eschariform lesions in such patients should provide a clinical clue to the presence of DF-2 infection.

Eur J Cancer Clin Oncol, 1985 Apr, 21(4), 439 - 45
Serum bactericidal activity as a therapeutic guide in severely granulocytopenic patients with gram-negative septicemia; Martino P et al.; The peak and trough levels of bactericidal activity of the serum of 74 severely granulocytopenic patients (less than or equal to 500 polymorphonucleates per microliter) with hematologic malignancies and Gram-negative septicemia were measured using the patient's infectious organism and serum containing the given antibiotics . When the peak titer of bactericidal activity in the serum was greater than 1:8 the septicemia was cured in more than 90% of the cases . However, in order to achieve a satisfactory rate of cure, patients with less than 100 polymorphonucleates/microliter required higher peak levels than patients with 100-500 polymorphonucleates/microliter . Serum bactericidal activity was influenced by the in vitro susceptibility of the offending pathogen and by the presence of in vitro synergism between the given antibiotics . These two variables showed a correlation with the clinical outcome that proved to be increasing with the degree of granulocytopenia . Furthermore, synergistic combination of the antibiotics appeared essential when the in vitro susceptibility shown by the offending pathogen was moderate . These data suggest (i) that determination of the bactericidal activity of the serum may prove to be a useful method to predict the clinical outcome in severely granulocytopenic patients with Gram-negative septicemia; and (ii) under the same conditions, antibiotic combinations that have demonstrable in vitro synergy against the offending pathogen should be given the utmost consideration.

Eur J Clin Microbiol, 1985 Apr, 4(2), 108 - 12
Evaluation of pertinent parameters of a new identification system for non-enteric gram-negative rods; von Graevenitz A et al.; The API 20 NE (Rapid NFT) system for identification of non-enteric aerobic gram-negative rods was evaluated using 431 strains of species covered by the analytical profile index and previously identified by conventional methods . In an analysis of several parameters, 343 (80%) of the strains were identified correctly with good to excellent probabilities of positive reactions (80.0-99.9%), 190 (55%) of these within 24 h; 74 (17%) yielded primary identifications with lower probabilities . Only 3% of the strains could not be correctly identified . The system is very satisfactory for routine use, but could be improved by modified pre-testing of the strains and by including more species in the profile index.

Nippon Geka Gakkai Zasshi, 1985 Apr, 86(4), 381 - 93
{Hyperdynamic state induced by continuous infusion of E . coli endotoxin in dogs}; Wu TT; It is well established that septic shock is caused by Gram-negative endotoxemia, and is characterized by hyperdynamic state being observed in the early stage of shock . This study is designed to elucidate the hemodynamic and metabolic changes induced by continuous infusion of small amount of endotoxin in the dog, with special reference to the existence of hyperdynamic state . Seven mongrel dogs were continuously infused with 10 micrograms/kg/day of E.coli endotoxin for 4 days under fasted, awake condition . All animals revealed mild hypodynamic state with severe catabolic responses in protein-energy metabolism . In contrast, 11 dogs infused with 36kcal/kg/day of glucose and 10 micrograms/kg/day of endotoxin showed marked increase (+30%) in cardiac index, and decrease in mean arterial pressure as well as total peripheral resistance, indicating the presence of hyperdynamic state . Metabolic studies indicate mild catabolic response . Administration of fat emulsion instead of isocaloric glucose has resulted transient hyperdynamic state, but failed to maintain this condition . In conclusion, it is possible to produce hyperdynamic state similar to that seen in septic shock by continuous administration of small amount of endotoxin in the dog, provided proper amount of glucose is supplied.

J Biol Response Mod, 1985 Apr, 4(2), 169 - 84
Biological effects of White-type polysaccharides of gram-negative bacteria; Rothman J et al.; The White-type polysaccharide (WPS), often called Freeman polysaccharide, was obtained by hydrolyzing gram-negative bacteria in 0.2 N acetic acid at 100 degrees C for 2 h . The crude product contained partially degraded O-antigens as well as other components that were active as immune adjuvants, enhancers of macrophage cytotoxicity of tumor target cells, and inducers of osteoclastic bone resorption . The same WPS preparation augmented the tumor cytotoxicity of normal mouse spleen cells and slightly retarded the take of L1210 leukemia in mice . The WPS preparations generated colony-stimulating factor in mice but were not active in lymphoproliferative (mitogenicity) tests . Chemical analyses of the WPS preparations did not detect the presence of components characteristic of the lipid moiety of endotoxins . The WPS samples were also negative in biological assays of endotoxicity such as local Shwartzman and toxicity tests . These findings indicate that gram-negative bacteria contain nonendotoxin components that are potent modifiers of some biological responses.

Oral Surg Oral Med Oral Pathol, 1985 Apr, 59(4), 336 - 9
The presentation and complications of odontogenic septic shock . Report of a case; Quinn P et al.; Although most odontogenic infections spread locally to adjacent fascial spaces, usually contiguous with the offending odontogenic pathosis, occasionally such an infection can spread rapidly across the midline and appear on the opposite side of the face and neck . Debilitated chronic alcohol abusers who are nutritionally deficient are more likely to develop serious life-threatening infections, either through serious airway involvement as seen in Ludwig's angina or manifest as a gram-negative septicemia with life-threatening shock and even cardiac arrest, than the usual dental patient with cellulitis . Early recognition through a high index of suspicion and vigorous monitoring will pick up the initial manifestation of toxic shock, as noted in this case report.

J Clin Hosp Pharm, 1985 Mar, 10(1), 95 - 100
Microbial contamination of non-sterile pharmaceutical products made in hospitals in the North East Thames Regional Health Authority; Baird RM; Non-sterile pharmaceutical products made in nine hospitals in the North East Thames Regional Health Authority were examined for microbial contamination as part of a quality assurance programme . The incidence of contamination was found to vary between hospitals, but in all cases improvements were seen during the 3-year monitoring period in the microbiological quality of products made in individual hospitals . The most common isolates were aerobic spore bearers--usually less than 10(2) colony forming units/g or ml . Gram-negative isolates were found in at least one product from each hospital . Pseudomonas spp . were isolated from several samples of peppermint water.

J Clin Pharmacol, 1985 Mar, 25(2), 67 - 81
Therapeutic initiatives for the avoidance of aminoglycoside toxicity; Whelton A; Aminoglycosides continue to be indispensable in the management of serious and life-threatening aerobic gram-negative infections . On an annual basis in the United States, they are used in the management of four million patients . Despite their clinical utility, they continue to manifest a high profile of toxic side effects such as nephrotoxicity and ototoxicity with the rare occurrence of neuromuscular toxicity . Animal experimental models have been invaluable in elucidating the pathophysiologic mechanisms by which aminoglycosides damage the kidney and the inner ear . However, it is from clinical therapeutic experience and prospective clinical trials that we have been able to solidly define the risk factors that accentuate the development of aminoglycoside-related toxicity . The clinical toxicity of these agents can be kept to a minimum by ensuring the use of an appropriate dose, for periods of time not exceeding nine to ten days, in a well-hydrated, normokalemic patient . Special subpopulation groups such as the elderly, the obese, those with preexisting renal disease, or patients who need the concurrent use of other nephrotoxins, require special care in the monitoring of their aminoglycoside therapy to ensure a safe and effective clinical outcome.

J Surg Res, 1985 Mar, 38(3), 298 - 304
Metabolic effects of pretreatment with Escherichia coli J5 antiserum on guinea pig gram-negative bacterial sepsis; Dunn DL et al.; Antiserum raised to a rough mutant Escherichia coli, termed J5 (anti-J5 RS), protected against lethal gram-negative bacterial sepsis in a guinea pig model when animals were pretreated with both antiserum and heparin . This same model was used to examine and compare the effects of pretreatment with anti-J5 RS, normal rabbit serum (NRS), or saline, each +/- heparin on physiologic and metabolic parameters during a septic insult . Results demonstrated that leukopenia and thrombocytopenia occurred to a similar degree in all pretreatment groups; no significant leuko- or thrombostasis was noted on examination of histologic specimens; complement activation was maximal in those animals receiving anti-J5 RS alone without heparin; the most abnormal amino acid profile was present in the NRS + heparin group; and only the anti-J5 RS + heparin group did not develop glomerular lesions indicative of disseminated intravascular coagulation . A complement-mediated cell aggregation-type injury does not appear to occur in this model . It is hypothesized that both excessive complement and coagulation system activation occur after bacterial challenge when antibody directed against the bacteria is present (anti-J5 RS) leading to antigen-antibody complex formation and complement and coagulation cascade activation . Heparin may block either or both these cascade systems allowing enhanced, antibody-mediated opsonization and clearance of blood-borne bacteria, thus preventing end-organ alterations and organ failure during sepsis when combined with anti-J5 RS.

Am J Obstet Gynecol, 1985 Mar 1, 151(5), 671 - 4
Endotoxemia in the neonatal lamb; O'Brien WF et al.; Although gram-negative sepsis is a major cause of neonatal morbidity and mortality, our understanding of endotoxemia in the neonate has been hampered by the lack of experimental models . Previous studies have suggested neonatal hyporesponsiveness to endotoxin . We studied unanesthetized neonatal lambs which had been exposed to the environment prior to study . These animals demonstrated the classic early phase changes of endotoxemia including pulmonary hypertension which was dependent upon prostanoid production . This model allows further studies of endotoxemia in the neonate.

Med Clin North Am, 1985 Mar, 69(2), 243 - 56
Gram-negative bacillary meningitis; LeFrock JL et al.; The incidence of gram-negative bacillary meningitis has increased significantly in the past two decades . Approximately two thirds of all reported cases have occurred after neurosurgical procedures . With the development of the newer cephalosporins, the overall mortality rate has decreased from 40 to 80 per cent to 10 to 20 per cent.

J Periodontol, 1985 Mar, 56(3), 127 - 31
Transmission and colonization of Actinobacillus actinomycetemcomitans in localized juvenile periodontitis patients; Christersson LA et al.; Actinobacillus actinomycetemcomitans is a Gram-negative oral microorganism, which has been implicated in the etiology of localized juvenile periodontitis and in severe medical infections such as bacterial endocarditis . This study evaluated the ability of periodontal probes to transmit A actinomycetemcomitans from juvenile periodontitis lesions to healthy gingival sulci in the same patient . Localized juvenile periodontitis patients exhibiting first molar and incisor alveolar bone loss and with large numbers of A actinomycetemcomitans in deep periodontal pockets were included in this study . A periodontal probe was inserted into periodontal pockets of 6 mm or greater depth . The probe was then placed into a healthy gingival sulcus of 3 mm or less, in the same subject . Fifty-five transfers by probing were made and A actinomycetemcomitans in both the donor and recipient sites was assessed by a selective culture technique . The results indicate that periodontal probes can become contaminated with A actinomycetemcomitans from juvenile periodontitis lesions during routine dental examinations and can transfer this microorganism from infected to previously uninfected sites . However, A actinomycetemcomitans inoculated into the healthy gingival sulci did not permanently colonize these sites since the organisms were eliminated within 3 weeks.

Proc Natl Acad Sci U S A, 1985 Mar, 82(6), 1790 - 4
Protection against gram-negative bacteremia and endotoxemia with human monoclonal IgM antibodies; Teng NN et al.; Hybridomas producing human monoclonal IgM antibodies (mAbs) against bacterial lipopolysaccharide (LPS) were generated by fusion of B lymphocytes from sensitized human spleen with heteromyeloma cells . The splenocytes were from patients undergoing splenectomy during staging for Hodgkin disease after vaccination with the J5 mutant of Escherichia coli, which is deficient in O antigenic side chains . This deficiency exposes the core oligosaccharide, common to LPS of all Gram-negative bacteria . The mAbs cross-reacted strongly with endotoxins from a wide range of unrelated species of Gram-negative bacteria . The mAbs also gave strong protection against LPS in the dermal Shwartzman reaction and against lethal Gram-negative bacteremia in mice . These findings indicate that monoclonal IgM against LPS endotoxin can neutralize its toxicity in vivo and might be valuable for treatment of patients with Gram-negative bacteremia . Analysis of one of the hybridoma clones, A6(H4C5), showed that the IgM mAb is directed against the covalently bound lipid A, which represents the most conservative and least variable structural element of LPS.

Plasmid, 1985 Mar, 13(2), 149 - 53
Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression; Ditta G et al.; Derivatives of plasmid pRK290 that are useful for cloning and for analyzing gene expression in a wide variety of Gram-negative bacteria are described . A smaller broad host range plasmid derived from RK2, with properties similar to that of pRK290, is also described.

J Bacteriol, 1985 Mar, 161(3), 955 - 62
Construction of broad-host-range cosmid cloning vectors: identification of genes necessary for growth of Methylobacterium organophilum on methanol; Allen LN et al.; Four new cloning vectors have been constructed from the broad-host-range cloning vector pRK290 . These vectors, pLA2901, pLA2905, pLA2910, and pLA2917, confer resistance to kanamycin and tetracycline . The latter two are cosmid derivatives of pLA2901 . The new vectors can be mobilized into, and are stably maintained in, a variety of gram-negative bacteria . A Sau3A genomic bank of Methylobacterium organophilum strain xx DNA has been constructed in pLA2917, and complementation analysis, with a variety of mutants unable to grow on methanol, revealed at least five separate regions necessary for growth on methanol . Complementation analysis and Tn5 mutagenesis data suggest that at least three genes are responsible for expression of active methanol dehydrogenase.

Minerva Med, 1985 Feb 25, 76(7), 261 - 8
{Exogenous infections and environmental flora . Criteria for the characterization of bacteria and parameters influencing their spread}; Pessione E et al.; An assessment was made of environmental micro-organisms isolated in the University of Turin Intensive Surgical Care Unit and Emergency Surgery Department to determine the possible exogenous origin of post-operative infections . The enzymatic characters and antibiotypes were determined for some species, and subsequently compared with similar characters of the "in vivo" isolated flora . A monthly disinfection method based on comparison among three antiseptic agents (chlorexidine, formaldehyde and formotetronium) considerably reduced the flora . The data obtained, while showing that most of the infections observed were endogenous, also provided indications on the way in which the spread of certain Gram-negative germs, often involved in post-operative and post-traumatic infections, could be contained.

J Immunol Methods, 1985 Feb 11, 76(2), 299 - 305
Immunoenzymatic analysis by monoclonal antibodies of bacterial lipopolysaccharides after transfer to nitrocellulose; Sidberry H et al.; A rapid, sensitive immunoenzymatic technique for the analysis of lipopolysaccharide (LPS) from gram-negative bacteria using monoclonal antibodies is described . After separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the LPS was either stained with silver or electrophoretically transferred to nitrocellulose . After reaction with anti-LPS monoclonal antibodies, the transferred antigens were visualized by reaction with alkaline-phosphatase-labelled anti-mouse antibodies and a substrate containing naphthol phosphoric acid and Fast Red.

Am J Med, 1985 Feb 8, 78(2A), 31 - 3
Aztreonam therapy for gram-negative pneumonia; Greenberg RN et al.; Nineteen patients with community- or hospital-acquired gram-negative pneumonia were treated with aztreonam . Other gram-negative antibiotics were withheld . Thirteen patients (68 percent) had clinical cures and 15 (79 percent) had microbiologic cures with aztreonam . No adverse reactions or drug toxicity occurred in this population.

Chemioterapia, 1985 Feb, 4(1), 24 - 7
Mechanisms of resistance to beta-lactam antibiotics; Wiedemann B et al.; The complex mechanism of resistance towards beta-lactam antibiotics in Gram-negative bacteria can be described by a simple equation in which the hydrolytic activity of the enzyme, the affinity to the penicillin binding protein in competition to the affinity to the beta-lactamase, the quantity of the enzyme and the ability of the drug to penetrate the outer membrane are considered . Depending on the characteristics of the drug, the beta-lactamase, and the microorganisms, the hydrolysis of the drug, its binding, or the reduced penetration can be the predominant factors.

Appl Environ Microbiol, 1985 Feb, 49(2), 305 - 9
Legionella anisa: a new species of Legionella isolated from potable waters and a cooling tower; Gorman GW et al.; Between March 1980 and June 1981, five strains of Legionella-like organisms were isolated from water . Four were recovered from potable water collected from hospitals in Chicago, Ill., and Los Angeles, Calif., during outbreaks of nosocomial legionellosis . The fifth strain was isolated from water collected from an industrial cooling tower in Jamestown, N.Y . The strains exhibited biochemical reactions typical of Legionella species and were gram-negative motile rods which grew on buffered charcoal-yeast extract agar but not on blood agar, required cysteine, and were catalase positive, urease negative, nitrate negative, hippurate negative, and nonfermentative . All strains were positive for oxidase and beta-lactamase and produced a brown, diffusible pigment . Of the five strains, four exhibited blue-white autofluorescence under long-wavelength UV light . The fatty-acid composition and ubiquinone content of these strains were consistent with those of other Legionella species . Direct fluorescent-antibody examination of the five strains with conjugates to previously described Legionella species demonstrated no cross-reactions except with the conjugates to L . longbeachae serogroup 2 and L . bozemanii serogroup 2 . Four strains gave a 4+ reaction to the L . longbeachae serogroup 2 conjugate and the fifth strain gave a 1+ reaction . Each of the five strains gave a 4+ reaction with the conjugate to L . bozemanii serogroup 2 . DNAs from the five strains were highly related (84 to 99%) and showed 5 to 57% relatedness to other Legionella species . These strains constitute a new species in the genus Legionella, and the name Legionella anisa sp . nov . is proposed . The type strain of L . anisa is WA-316-C3 (ATCC 35292).

Am Rev Respir Dis, 1985 Feb, 131(2), 209 - 13
Endotoxin in cotton dust and respiratory function decrement among cotton workers in an experimental cardroom; Rylander R et al.; To study the various reactions of gram-negative bacteria and their endotoxins observed in workers exposed to cotton dust, experiments were undertaken where cotton mill workers carded cottons from different geographic locations, each containing different amounts of bacterial endotoxins . Exposure was determined as the vertical elutriator dust and endotoxin levels . Measurements were made of the forced expiratory volume in one second (FEV1) and the number of blood neutrophils before and after work, and the prevalence of symptoms of byssinosis was recorded . There was a significant correlation between the vertical elutriator endotoxin levels and the group mean changes in FEV1 (p less than 0.01) but no correlation between vertical elutriator dust levels and changes in FEV1 . There was also a dose-response relationship between the endotoxin levels and the presence of symptoms of byssinosis in the exposed workers as well as between endotoxin levels and an increase in blood neutrophils . This observation supports findings from several previous studies and suggests that endotoxin triggers the mechanisms responsible for the decrease in respiratory function in the byssinosis syndrome . Other constituents of cotton dust could also be of importance for this reaction.

Mol Gen Mikrobiol Virusol, 1985 Feb, (2), 3 - 14
{Development of a system vector-host in methylotrophic bacteria}; Tsygankov IuD et al.; Cloning of methylotropic and other Gram negative bacteria's genes was performed using vectors derived from IncP4 plasmids . Plasmids, such s RSF1010 are 8.8 kb in length, have a high copy number and broad host range and can be mobilized efficiently by a number of conjugative plasmids . IncP4 plasmids have relatively few restriction enzyme's targets suitable for cloning . In this paper the construction of versatile and special purpose IncP4 vectors available for cloning DNA into broad range of bacterial species are described . The seria of versatile vectors involves the transposon containing plasmid and two-replicon vectors.In genetic construction of special vector for direct cloning of restriction fragments the genetic regulation elements of Tn 1 were used . On the base of IncP4 replicon special vectors for construction of bank genes (cosmids) and the vectors for cloning of regulation sequence were also constructed.

Immunobiology, 1985 Feb, 169(1), 1 - 10
Serum anti-lipid A antibodies in multiple myeloma and Waldenström's macroglobulinaemia; Fink PC et al.; Anti-lipid A antibodies were determined in sera from 38 patients with IgG multiple myeloma, 33 patients with IgA multiple myeloma and 38 patients with Waldenstrom's macroglobulinaemia using an enzyme-linked immunosorbent assay (ELISA) . Compared to 34 healthy adults, significantly (p less than 0.025, p less than 0.005, p less than 0.0025) lower serum anti-lipid A antibody levels were measured for the respective patient groups . Low anti-lipid A antibody levels correlated with a higher infection rate with gram-negative bacteria in patients with monoclonal B-cell malignancies . The highest infection rate was seen in patients with simultaneous low anti-lipid A levels and secondary antibody deficiency.

Immunopharmacology, 1985 Feb, 9(1), 45 - 52
Synthesis of PAF-acether and blood volume changes in gram-negative sepsis; Inarrea P et al.; Gram-negative sepsis was induced in rats by intraperitoneal injection of Escherichia coli . The development of bacterial peritonitis and septicemia was monitored by counting the number of peritoneal cells and by performing cultures of blood samples . Mortality reached a 50% rate when rats were injected with 2 X 10(8) colony-forming units . Rats injected with the doses of bacteria which induced mortality showed a time- and dose-dependent increase of vascular permeability as judged by the presence of abundant peritoneal exudate and by the depletion of the circulating volume . In order to know whether the generation of PAF-acether could be involved in the development of the permeability changes, the formation of this mediator was measured in the peritoneal cells and spleen of animals at different times and in response to different doses of E . coli . Significant amounts of PAF-acether could be obtained preceding the development of blood volume depletion in response to the injection of doses of E . coli which induced both mortality and the development of permeability . These data suggest that PAF-acether might be one of the inflammatory mediators involved in the pathogenesis of the hemodynamic changes observed in endotoxemia.

Antimicrob Agents Chemother, 1985 Feb, 27(2), 217 - 9
Lack of evidence for interaction between tobramycin and shock in their effect on renal function; Ambinder RF et al.; We sought to determine whether there was an interaction between aminoglycoside use and shock in their effect on renal function among seriously ill patients suspected of having gram-negative sepsis . Serial serum creatinine determinations were used to estimate changes in creatinine clearance rates in 179 patients entered onto a prospective randomized trial of tobramycin-nafcillin versus cefotaxime . A 25% decline in estimated creatinine clearance was considered to be clinically important . Univariate chi-square analysis showed that both shock (P less than 0.01) and tobramycin use (P less than 0.001) were independently associated with decline in estimated creatinine clearance . A two-way analysis of variance showed that both shock (F = 10.44, P less than 0.01) and tobramycin use (F = 42.6, P less than 0.001) continued to be significantly associated with renal dysfunction in the presence of each other, but there was no significant interaction between shock and tobramycin in their effect (F = 0.62, P less than 0.43) . A multiple logistic regression with an interaction term representing the occurrence of shock and tobramycin use simultaneously yielded similar results . Our study provided no analytic evidence supporting the existence of an interaction between shock and aminoglycoside use in their effect on renal function.

Schweiz Med Wochenschr, 1985 Jan 19, 115(3), 93 - 6
{Aminoglycoside levels in bronchial secretions}; Mombelli G; Aminoglycosides are of limited clinical efficacy in gram-negative bacillary pneumonia, although they are commonly employed to treat this infection . This poor efficacy has been related in part to host factors (abnormalities in the lung, immunocompromise) and in part to pharmacologic factors . In particular, aminoglycoside levels in bronchial secretions are often borderline or inadequate in relation to the minimal inhibitory concentrations for most gram-negative strains . The low aminoglycoside concentrations result from poor penetrance into the respiratory tract or from local inactivation of these drugs, but basically reflect their low therapeutic-to-toxic ratio . The endotracheal injection of aminoglycosides resulted in high bactericidal activity within the bronchial lumen and in increased clinical efficacy, without increasing systemic toxicity . In view of the potential dangers of topical antibiotics, however, endotracheal treatment should be confined to selected patients.

Circ Shock, 1985, 17(4), 301 - 11
Effects of toxic and nontoxic endotoxin derivatives on glucose kinetics; Lang CH et al.; Although the polysaccharide portion of bacterial endotoxins (ET) and other components of the bacterial cell wall are known to possess biological activity, the toxic effects produced by ET have been attributed mostly to the lipid-A component . The present study examined the influence of selected gram-negative cell wall components on in vivo carbohydrate homeostasis . Chronically catheterized conscious rats were injected with either ET, lipid A, the White polysaccharide supernatant (WPS-S; polysaccharide-rich and lipid-A-free) or the WPS-precipitate (WPS-P; rich in cell wall components and lipid-A-free) at sublethal doses of 100, 10, and 1 microgram/100 g . The acute hypotensive response to ET and lipid A were similar, while the WPS-P induced a smaller reduction in pressure . Endotoxin, lipid A, or the WPS-P (100 and 10 micrograms) all produced hyperglycemia . Hyperlactacidemia was evident in rats receiving ET, lipid A, and WPS-P; ET-treated animals exhibited the highest lactate concentrations . At the highest doses used, these three fractions increased the rate of glucose appearance . The polysaccharide-rich WPS-supernatant elicited no significant alterations in any of the variables studied . These results indicate that cell wall components of gram-negative bacteria, other than the lipid A of endotoxin, induce changes in carbohydrate metabolism that are similar to those produced by the toxic, lipid-A component of endotoxin.

Circ Shock, 1985, 16(2), 195 - 203
Effect of methylprednisolone and of ibuprofen, a nonsteroidal antiinflammatory agent, on bronchoalveolar inflammation following endotoxemia; Rinaldo JE et al.; Septicemia by gram-negative organisms is a common cause of the adult respiratory distress syndrome (ARDS) . The role of neutrophils in causing parenchymal lung damage in ARDS has recently been emphasized . A single intraperitoneal injection of Escherichia coli endotoxin in rats causes acute neutrophil alveolitis similar to that of ARDS . We studied the ability of pretreatment with either ibuprofen (IBU) or methylprednisolone (MP) to ablate directly the alveolar inflammatory response to endotoxin in the rat model . To compare the severity of inflammation, we quantified inflammatory cell recovery by whole lung lavage 24 hours after injection of endotoxin, the time point at which neutrophil alveolitis due to endotoxin is most intense . Pretreatment with a single dose of IBU 3.75 mg/kg prior to endotoxin injections was associated with a significant increase in the total number of inflammatory cells, and in both the percentage and the absolute number of neutrophils recovered from the lung, despite significantly decreasing the plasma level of thromboxane B2, which increased 10-fold after endotoxin . Paradoxically, IBU 30 mg/kg significantly decreased the intensity of neutrophil alveolitis . MP 30 mg/kg had no effect on recovery of inflammatory cells from the lung by bronchoalveolar lavage following endotoxin . Cyclooxygenase inhibitors such as ibuprofen may cause a dose-dependent biphasic effect on lung inflammation following endotoxin: enhancement of inflammation at a low dose and suppression of inflammation at a high dose.

Prog Clin Biol Res, 1985, 189, 347 - 67
The importance of intestinal endotoxins in liver disease; Nolan JP et al.; The development and wider use of the Limulus amoebocyte lysate assay (LAL) for the detection of endotoxin, has led to renewed interest in the link between gut-derived endotoxin, liver injury and the extra-hepatic manifestations of clinical liver disease . The concept that endotoxemia may occur and be harmful in clinical states without the concomitant presence of Gram-negative bacteria is a relatively recent one and has been most intensively studied in the context of liver injury . Since the liver stands between the gut and the systemic circulation, it has been postulated that failure to detoxify endotoxins absorbed into the portal circulation after hepatic injury might lead to further liver damage and escape of this toxic material into the general circulation . The present review will update the status of this hypothesis, discuss mechanisms proposed for the damage and present evidence for the association in animal models . Finally, studies both positive and negative, will be cited that have used the LAL to detect endotoxemia and its consequences in human liver disease.

Prog Clin Biol Res, 1985, 189, 329 - 45
Plasma Limulus gelation assay in infants and children: correlation with gram negative bacterial infection and evidence for "intestinal endotoxemia"; Cooperstock M et al.; Plasma limulus gelation assays for gram negative bacterial endotoxemia were performed in 103 infants and children with evaluable clinical findings . A strong association between gram negative infection and positive assays was found, provided patients with gastrointestinal disorders were considered separately . Ten of 11 patients (91%) with gram negative bacteremia (p less than .001 compared with controls) and five of 10 (50%) with focal gram negative bacterial infections (p = .002) had a positive assay . Only 1/22 (5%) of those classified as having other forms of infection-like illness with negative cultures, and none of 20 considered to have neither infection nor gastrointestinal disturbances had a positive assay . However, 8/24 (33%) of those with major gastrointestinal disturbances had a positive test (p = .003) . This association supports the possibility that endotoxins produced by indigenous gram negative bacteria in the gut may reach the circulation during states of intestinal disturbance (intestinal endotoxemia) . Individual cases suggest several different possible mechanisms which could account for such an occurrence . Rigorous proof of this concept must await the development of confirmatory methodology.

Prog Clin Biol Res, 1985, 189, 169 - 80
Parallel line assays of endotoxins with the LAL chromogenic substrate method; van Noordwijk J; All endotoxins from the gram negative microorganisms studied sofar contain lipid A, which is considered to be responsible for the biological activity . Endotoxins may therefore react in a similar way in quantitative assays . The varying non-lipid A part of different endotoxins may potentiate or reduce the activity of the identical lipid A part without changing the slope of the log-dose vs . response curve of these endotoxins . To test whether 13 endotoxins from different microorganisms react similarly with amoebocyte lysate (LAL) log-dose response curves were made using a chromogenic substrate method . A balanced incomplete block design with 13 blocks and four replications of each endotoxin was applied . A group of eight endotoxins with a common slope was defined with the Student-Newman-Keuls procedure . Two independent potency estimates of these endotoxins were in good agreement.

Prog Clin Biol Res, 1985, 189, 117 - 37
Optimalization of a chromogenic assay for endotoxin in blood; Sturk A et al.; Early detection of Gram-negative septicemia or endotoxemia may become feasible with sensitive and reliable endotoxin (LPS) measurements . We recently published an assay for LPS in blood, based upon the LPS dependent activation of Limulus amebocyte lysate (LAL) and the subsequent chromogenic measurement of the activated enzyme(s) . Inhibitors and activated clotting factors potentially interfering in the assay were removed by dilution and heating . In the present study we describe the further improvement of the assay . Optimal conditions include: blood anticoagulated with 30 I.U./ml heparin; dilution of the platelet-rich plasma (PRP) in water; 5 min . heating at 75 degrees C; 15 mM Mg2+, 1.5 mM Ca2+, 125 mM Na+, 50 mM TRIS, pH 8.5 in the LAL activation step; substrate step without extra addition of Ca2+, Mg2+, or Na+, but in the presence of 50 mM TRIS at pH = 9.5 . Under those optimal conditions less than 10 pg LPS per ml blood (PRP) can easily be detected . Prospective clinical trials are presently envisaged to investigate the clinical usefulness of this extremely sensitive LPS assay.

Drugs, 1985, 29 Suppl 5, 140 - 5
Biliary concentrations of temocillin; Poston GJ et al.; Temocillin is a new semisynthetic beta-lactam antibiotic with a spectrum of activity against most aerobic Gram-negative bacteria . We investigated the biliary excretion of temocillin after administration of 1g either intravenously or intramuscularly to 15 patients with normal liver and renal function who were undergoing elective cholecystectomy for gallstones . Very high concentrations of temocillin (up to 8 times the corresponding serum concentrations) were found in common bile duct bile and gallbladder bile of functioning gallbladders 1 to 2 hours after administration . However, no temocillin was detected in the bile of non-functioning gallbladders . Concentrations of temocillin were significantly higher in common bile duct bile (p less than 0.01) and gallbladder bile (p less than 0.05) following intravenous administration, and there was also a significant correlation (p less than 0.05) between common bile duct bile and corresponding serum concentrations for both routes of administration . Temocillin 1g readily achieved therapeutic concentrations in bile, and may prove a useful agent in the prophylaxis and management of Gram-negative biliary infections.

Drugs, 1985, 29 Suppl 5, 118 - 21
Clinical efficacy, tolerance and pharmacokinetics of temocillin in patients with respiratory tract infections; Legge JS et al.; In this study 13 patients with a history of chronic respiratory disease were treated with temocillin for severe infection of the lower respiratory tract . Eleven patients were clinically improved or cured at the end of treatment and 4 remained free of infection at follow up . However, the 2 remaining patients failed to respond clinically and bacteriologically to temocillin . Blood and sputum pharmacokinetics were studied in patients receiving temocillin at 12-hourly intervals in doses of either 1g by intravenous bolus or 2g by intravenous infusion . On the first day of treatment the mean peak serum concentration was 102 mg/L for the 1g dose and 220 mg/L for the 2g dose, falling to 11 mg/L (1g) and 18.8 mg/L (2g) at 12 hours . Similar results were recorded on the last day of treatment . Temocillin concentrations in the sputum ranged from 1.5 to 2.8 mg/L, with no difference being found in the sputum concentrations of the 2 doses . Temocillin was well tolerated and was found to be an effective treatment for Gram-negative bacterial infections of the lower respiratory tract.

Drugs, 1985, 29 Suppl 5, 103 - 5
Temocillin . Summary of safety studies; Cockburn A et al.; Temocillin is a novel injectable beta-lactam antibiotic designed for parenteral use . It is active against the majority of Gram-negative bacteria and is stable to a wide range of beta-lactamases . Disposition and metabolic studies on temocillin in animals and man demonstrate that the drug is well distributed throughout the body tissues and will cross the placenta . The major route of elimination is via the kidney (89%) where the drug is excreted unchanged as parent compound, mainly by the glomerular filtration of unbound temocillin . A full toxicological safety evaluation programme has been completed and includes acute, subacute and chronic toxicology in rat and dog, and reproductive and mutagenicity studies . Temocillin was well tolerated in animals at dosages of up to 1000 mg/kg and appears to be without any potential hazard for man in dosages which are well in excess of the proposed therapeutic dose.

Res Exp Med (Berl), 1985, 185(3), 181 - 93
Efficacy of immunoglobulins in gram-negative infections in experimentally induced peritonitis in the rat; Frick S et al.; The effect of adjuvantly administered, newly developed immunoglobulin preparations in combination with an antibiotic is investigated by means of an animal experimental model . The animal model was peritonitis which was induced by a left-open colotomy in the rat . Administration of a combination therapy of immunoglobulins and an antibiotic succeeded in reducing lethality and shock index (according to Staub {15}) significantly by up to 50% as compared to the untreated control group . Using sub-therapeutic dosage of the antibiotic (50% of the human equivalent dose) the synergistic effect of the immunoglobulins could be confirmed clearly . A single summation of the offered immunoglobulin preparations cannot achieve an improvement in the results . In the present examinations additional IgM-enrichment does not result in a further improvement over the usually normal immunoglobulin levels in the preparations.

Antibiot Chemother, 1985, 36, 103 - 10
Antibiotic uptake and the cytoplasmic membrane; Bryan LE; Aminoglycoside antibiotics are accumulated through a unique bacterial transport process . Knowledge of the process and how it can alter susceptibility to aminoglycoside antibiotics may be used to investigate mechanisms that might increase the activity of these compounds . It is possible that aminoglycoside activity could be increased by presenting more drug to the cytoplasmic membrane through alteration of the outer membrane of gram-negative bacteria by increasing transporter concentration or driving force in the form of the electrical potential, by decreasing the rate of enzymatic modification so that active drug can be transported, or by changing aminoglycosides to utilize alternative forms of transport and finally to increase ribosomal affinity for these drugs.

Circ Shock, 1985, 15(4), 247 - 59
The effect of hyperdynamic sepsis on myocardial performance; McDonough KH et al.; The reviewed studies support the contention that during the high flow or hyperdynamic phase of gram-negative septicemia, cardiac reserve is compromised because of intrinsic myocardial dysfunction . The latter is not referable to coronary hypoperfusion or peripheral pooling or decreased venous return . Although, under resting, nonstressed conditions, indices of myocardial function may appear normal or even elevated, a decreased reserve is evident when additional stress is imposed on the myocardium . Hearts removed from septic rats during the hyperdynamic stage and perfused in vitro (using the isolated perfused working heart preparation) showed a rightward and downward shift in work function curves, indicating a severe depression in cardiac function . Possible mechanisms for the observed dysfunction are discussed . No significant alterations in high energy phosphate production or substrate utilization were observed, indicating that altered myocardial metabolism is not likely to be a significant contributor to the dysfunction . Our results suggest that cardiac dysfunction is partially due to an elevation in the cytosolic calcium concentration which may slow the rate of ventricular relaxation . These studies emphasize that intrinsic cardiac function is depressed early during the course of the septic episode at a time that precedes the onset of circulatory shock.

Circ Shock, 1985, 15(4), 227 - 45
Isolated cardiac preparations: models of intrinsic myocardial dysfunction in circulatory shock; Parker JL et al.; Isolated cardiac preparations have been adapted for modeling intrinsic myocardial responses to circulatory shock syndromes independently of immediate influence from depressive or supportive constraints operative in the intact host . Left atrial and left ventricular (LV) papillary muscles and coronary-perfused hearts were removed from guinea pigs during development of Escherichia coli endotoxin shock . Preparations were then subjected to a battery of functional analyses under conditions of constant perfusate pH, pO2, pCO2, electrolyte and substrate concentrations, osmolality, and temperature . Evidence for contractile depression intrinsic to the myocardium itself was a consistent and reproducible finding in all three tissue models . The LV mechanical disadvantage of shock hearts was not correlated with changes in cardiac cycle length (beating frequency), active state duration (contraction-relaxation intervals), or tissue water content; neither was it surmounted by maximally effective increments in coronary flow, {Ca2+}o, or diastolic fiber length . Taken in concert, studies to date suggest that gram-negative endotoxin leads in some way to a reduction of intrinsic contractile reserves of the myocardium, and we have postulated that this change underlies the circulatory shock phase of endotoxicosis . The experimental approach embodied in these models may yield unique cardiodynamic interpretations that will allow the formulation of testable hypotheses about the pathogenesis and prevention of intrinsic cardiac complications of endotoxin and related shock forms.

Vet Med Nauki, 1985, 22(1), 36 - 9
{Rapid method for determining the drug resistance of gram-negative bacteria}; Pavlov A et al.; Comparative studies led to the working out of a new diffusion indicative method for the rapid determination of drug sensitivity in Gram-negative bacteria . It is a modification of the diffusion method, consisting in the use of an indicator medium, bromothymol blue in glucose agar, instead of the meat peptone agar or other routinely employed nutritive media . The new method makes it possible to record rapidly the drug sensitivity of bacteria--only for 4-6 hours (compared with 24 hours with the conventional method) . Besides, the new method is readily applicable, precise, distinctive and rapid.

Acta Neuropathol (Berl), 1985, 65(3-4), 344 - 6
"Pink spots" in formalin-fixed brains--poor fixation or bacterial byproduct?
Hedley-Whyte ET.
Circumscribed pink areas 0.3-1.0 cm in diameter seen in deeper parts of otherwise well-fixed brains are usually ascribed to inadequate fixation . Twenty-three patients with pink areas in their fixed brains had evidence of inflammation in at least one organ . Blood vessels in the pink areas contained bacteria . Postmortem cultures identified the organisms in 16 cases . Gram-negative bacteria were the most common . Sharply delineated pink areas in well-fixed brains correlate with postmortem intravascular growth of bacteria.

J Reprod Med, 1985 Jan, 30(1), 67 - 8
Capnocytophaga isolated from the endometrium as a cause of neonatal sepsis . A case report; Mercer LJ; Capnocytophaga, a genus of gram-negative, anaerobic organisms, was isolated from the endometrium of a postpartum woman and the blood of the neonate . This unusual organism, usually isolated from the oral cavity, is sensitive to most antibiotics except the aminoglycosides.

Arch Surg, 1985 Jan, 120(1), 99 - 103
Serum level monitoring of aminoglycoside antibiotics . Limitations in intensive care unit-related bacterial pneumonia; Flint LM et al.; Serum aminoglycoside assays have been accepted as useful methods of enhancing therapeutic efficacy in the treatment of intensive care unit-acquired pneumonia and in avoiding aminoglycoside nephrotoxicity . We prospectively studied 68 surgical patients with normal renal function and gram-negative bacterial pneumonia who were treated with aminoglycosides . Serum levels indicated subtherapeutic levels in 47 patients and verified optimum levels in 13 patients . Toxic trough levels developed in six patients and, despite immediate dosage adjustment, five patients suffered nephrotoxicity . Six additional patients also had nephrotoxicity . Five of these patients never had toxic peak or trough levels and rising trough levels developed in one patient after serum creatinine levels began to rise . We conclude that routine monitoring of serum levels effectively detects subtherapeutic antibiotic levels . This modality is useful for optimizing dosage schedules, but does not serve to predict or avoid nephrotoxicity in critically ill surgical patients.

Drugs Exp Clin Res, 1985, 11(6), 373 - 8
Behaviour of cefmenoxime towards beta-lactamases; Labia R et al.; Using several well-characterized beta-lactamases isolated from Gram-negative bacteria, the interactions of cefmenoxime, a new methoxy-imino-amino-2-thiazol cephalosporin were compared with those of cefotaxime, lamoxactam, cefoperazone and ceftazidime . On-line computerized microacidimetry allowed determination of the affinity of these compounds for the enzymes, which was characterized by Ki values . Microacidimetry showed poor interactions of cefmenoxime with penicillinase TEM-1 (low Vm, poor affinity) whereas it showed a high affinity for the cephalosporinases, as is also the case for cefotaxime or lamoxactam . Both cefmenoxime and cefotaxime showed relative susceptibility in Masuda's double disc technique.

Antibiot Med Biotekhnol, 1985 Jan, 30(1), 55 - 61
{Present-day aminoglycosides in the treatment of suppurative-septic diseases}; Lobuseva AN et al.; The therapeutic efficacy, tolerance and pharmacokinetics of sisomicin, gentamicin and a new dosage form of kanamycin sulfate as an ampoule solution for intravenous injection were studied clinically . High therapeutic efficacy of the above three aminoglycosides in the treatment of severe inflammatory infections caused by gram-negative bacteria was shown with the use of the adequate doses . Favourable results of the therapy estimated by the laboratory and clinical data were observed in 81-90 per cent of the patients.

Prog Biochem Pharmacol, 1985, 20, 26 - 37
Pulmonary dysfunction caused by diffuse lung inflammation . Roles of metabolites of arachidonic acid; Brigham KL; Gram-negative endotoxemia in chronically instrumented sheep causes diffuse lung inflammation . Pathophysiologic responses of the lung include marked changes in lung mechanics, pulmonary vasoconstriction, increased lung vascular permeability, and capillary endothelial injury . Over the course of the response to endotoxemia, cyclooxygenase and lipoxygenase products of arachidonic acid are released into lung lymph and the time course of the two classes of compounds is different . Thromboxane concentrations in lung lymph increase early, coincident with the most marked changes in lung mechanics and the most severe pulmonary hypertension . Concentrations of a prostacyclin metabolite also increase during this early phase of the endotoxin response but peak levels in lung lymph appear slightly later than that for thromboxane . Concentrations of these cyclooxygenase products subside as the later phase of increased vascular permeability develops . Lung lymph concentrations of two lipoxygenase products, 5- and 12-HETE, increase late in the endotoxin reaction, approximately coincident with physiologic evidence of increased lung vascular permeability . Neutrophil chemotactic activity appears in lung lymph early after endotoxin infusion and persists for several hours . Drugs which inhibit cyclooxygenase attenuate the early changes in lung mechanics and the early pulmonary hypertension after endotoxemia, but do not prevent the late phase increase in vascular permeability, suggesting that the net effect of endogenous generation of cyclooxygenase products explains the early constrictor phase of the reaction, but not the later capillary injury . However, neutrophil depletion prevents the early changes in lung mechanics without preventing the coincident pulmonary hypertension or the increase in lung lymph thromboxane concentrations which may indicate that the cell source of constrictor cyclooxygenase products mediating changes in lung mechanics is different than that for products causing pulmonary vasoconstriction . The specific role of prostacyclin in the lungs' response to endotoxemia is not clear . The temporal course of prostacyclin release suggests that it may play a role in moderating the pulmonary hypertension and changes in lung mechanics . The fact that neutrophil depletion, corticosteroids, and the antioxidant n-acetylcysteine diminish both the endotoxin-induced increase in lung vascular permeability and lung prostacyclin release may indicate that the release of prostacyclin is a response to endothelial injury.(ABSTRACT TRUNCATED AT 400 WORDS)

Arch Surg, 1985 Jan, 120(1), 50 - 3
Enhanced survival during murine gram-negative bacterial sepsis by use of a murine monoclonal antibody; Dunn DL et al.; We developed a murine monoclonal antibody (5B10 MAb) that reacted in vitro specifically to lipopolysaccharide (LPS) obtained from Escherichia coli 0111:B4 . Enzyme-linked immunosorbent assay (ELISA) titers to a variety of gram-negative bacterial whole cell and LPS antigens demonstrated that this antibody may react with the O antigen portion of 0111:B4 LPS . We then examined the ability of this antibody to protect mice in vivo against a challenge of either viable bacteria or purified LPS . One milligram of 5B10 MAb was administered intraperitoneally (IP) and protected against a lethal challenge of either viable E coli 0111:B4 or 0111:B4 LPS, but no other type of bacterial or LPS challenge . Protection occurred in an antibody dose-dependent manner, and as little as 0.01 mg of 5B10 MAb enhanced survival . We concluded that IP pretreatment with a single MAb would protect against lethal sepsis or endotoxemia in this animal model and that anti-LPS specificity was a sufficient condition for an antibody to protect during bacteremia, confirming the importance of LPS in the pathogenesis of gram-negative bacterial sepsis.

Boll Ist Sieroter Milan, 1985, 64(4), 255 - 61
Endotoxicity of newly recognized gram negative organism: a collective review; Fumarola D; A comprehensive picture of the in part personal investigations concerning an Endotoxin-like activity associated with some recently recognized Gram negative significant pathogens has been drawn . Suspensions of heat-killed organisms (or endotoxic lipopolysaccharide when available) have been tested using as markers of endotoxicity the Limulus assay and the skin Shwartzman reaction in rabbit . Some general considerations are finally set forth on the possible role of the surface structures in mediating (with other virulence factors) the damage in the host.

Circ Shock, 1985, 16(3), 241 - 52
Myocardial dysfunction in a nonlethal, nonshock model of chronic endotoxemia; Fish RE et al.; Escherichia coli endotoxin (ET) was administered to adult rats by continuous IV infusion from a subcutaneously implanted osmotic pump (Alzet) . Myocardial function was assessed after 6 and 30 hr of ET infusion and compared with control rats which received a saline infusion and were fasted to match the anorexia of ET rats . Cardiac output (CO) and coronary blood flow, measured by the radiolabeled microsphere method, and mean arterial blood pressure, heart rate, total peripheral resistance, and stroke volume, were determined in vivo . Treatment differences were limited to a 13% lower arterial pressure in ET rats after 30 hr of infusion . Myocardial function was evaluated in vitro in similarly treated rats with the isolated perfused working heart preparation; preload was altered by raising the left atrial filling pressure (LAFP) from 10 to 30 cm water . After both 6 and 30 hr of infusion, hearts from ET rats exhibited a significantly lower peak systolic pressure (PSP), CO, and coronary flow in response to increasing LAFP, and a greater oxygen consumption per unit of myocardial work (CO X PSP) . Reduced in vitro work performance of hearts from endotoxemic rats was demonstrated early in the course of chronic endotoxemia and in the absence of in vivo evidence of cardiac dysfunction . Myocardial dysfunction, masked in vivo by compensatory mechanisms used to maintain adequate cardiovascular function, may be an important feature in the pathogenesis of both experimental endotoxemia and clinical gram-negative sepsis.

Prog Clin Biol Res, 1985, 189, 387 - 403
Rapid diagnosis of genitourinary infections by use of the Limulus lysate amebocyte test; Jorgensen JH; The Limulus amebocyte lysate (LAL) assay has realized numerous clinical applications, including rapid diagnosis of significant Gram negative bacteriuria and gonorrhea . Several published studies have demonstrated that the LAL test detects significant Gram negative bacteriuria with greater than 90% accuracy and that the predictive value of a negative test (97%) is similar to that of other currently available screening methods for bacteriuria . The test has also succeeded when used for presumptive diagnosis of gonorrhea in men, although it has not been shown to be superior to Gram stain examination of urethral exudate . The LAL test appears to offer excellent sensitivity for examination of cervical secretions from women being screened for gonorrhea, although the specificity of the test appears low, especially when a low-risk population is being examined.

Adv Exp Med Biol, 1985, 184, 35 - 50
Killing of gram-negative bacteria by neutrophils: role of O2-independent system in intracellular killing and evidence of O2-dependent extracellular killing; Weiss J et al.; For many strains of these two bacterial species, ingested bacteria are efficiently killed by O2-independent mechanisms . Intracellular killing depends not on 2de novo generation of toxic products of the respiratory burst but rather on intracellular delivery to pre-existing cytotoxic proteins . The principal O2- independent bactericidal system toward these bacteria appears to be BPI which rapidly binds to ingested bacteria and whose discrete action closely resembles the initial lesions produced by the intact neutrophil . In addition, at least for one strain of E . coli (S15), extracellular bacteria can be killed in an O2- dependent fashion.

Ann Otolaryngol Chir Cervicofac, 1985, 102(3), 187 - 93
{Postoperative infection in ORL oncologic surgery . The choice of preventive antibiotherapy . Experience of the ORL Department of Hôpital Claude Bernard}; Gehanno P et al.; The incidence of infection after cervicofacial surgery for ENT cancer varied between 38 and 60% as a function of the type of operation, and prophylactic antibiotic therapy administered, in a series of 133 cases . Contamination was greatest after operations of longest duration and with the largest exposure of tissue . Previous cobalt therapy was an aggravating factor . The highest risk of infections complications resulted therefore from combined salvage operations and the use of a musculocutaneous flap . Among these infections, a purulent bronchorrhea was a frequent and potentially serious complication sometimes the cause of septicemia . A review of 4 different antibiotic therapy regimens showed the pre-operative administration failed to alter the outcome . Combined penicillin-5 imidazole treatment, effective in preventing local and regional anaerobic infection, resulted in secondary infection developing in about 56% of cases . It was not able, in fact, to cover under certain circumstances during prolonged surgery, the postoperative risk of inhalation of gram-negative anaerobic and aerobic bacteria form the buccal cavity . Short-term antibiotic therapy was totally ineffective . It is suggested that combined penicillin-5 imidazole be reserved for operations provoking little contamination of the tracheobronchial region, such as total laryngectomy and total pharyngo-laryngectomy, and that prophylactic therapy using an antibiotic with a wide spectrum against gram negative germs be administered prior to major buccopharyngeal operations with tracheotomy.

Ther Drug Monit, 1985, 7(1), 12 - 25
An overview of amikacin; Ristuccia AM et al.; Amikacin, a semisynthetic analog of kanamycin, is very active against most gram-negative bacteria including gentamicin- and tobramycin-resistant strains . The effectiveness of amikacin in the treatment of serious gram-negative bacillary infections is well documented . Due to its resistance to inactivating enzymes, it is the aminoglycoside of choice for the treatment of known or suspected serious gram-negative infections caused by organisms resistant to gentamicin or tobramycin . Amikacin should be part of an empiric antibiotic regimen for the therapy of suspected sepsis in febrile, leukopenic immunocompromised hosts since it exhibits enhanced activity against the organisms most frequently encountered in this patient population . High response rates have been reported with the use of amikacin combined with beta-lactam antibiotics in immunocompromised or granulocytopenic patients . It exhibits impressive in vitro synergy against aminoglycoside-sensitive and -resistant organisms when used in combination with the new acylureidopenicillins and third-generation cephalosporins . Amikacin has the advantage of being the aminoglycoside least inactivated by the semisynthetic penicillins . Amikacin achieves high and predictable serum concentrations and has a favorable therapeutic index . Its potential for nephrotoxicity and ototoxicity is not significantly different than that encountered with gentamicin or tobramycin . Amikacin appears to be the preferred aminoglycoside for use at the present time because of its activity against gentamicin- and tobramycin-resistant organisms, its low resistance potential, its relative low degree of inactivation by the semisynthetic penicillins, and its superior pharmacokinetic profile.

Infection, 1985 Jan-Feb, 13(1), 8 - 11
Opsonic activity of the alternative complement pathway in infected human intra-abdominal fluid; Lam C et al.; The complement-mediated opsonic activity (CMOA) in intra-abdominal exudates collected during laparotomy from peritonitis patients was examined by an in vitro phagocytic bactericidal assay . It was found that infected intra-abdominal exudates barely promoted detectable killing of Escherichia coli 01 by polymorphonuclear leukocytes . Only the minority of bacteria recovered by differential centrifugation from the infected exudates had C3 on their surfaces . Such bacteria were not optimally opsonized in vivo, whereas they became fully coated with C3 during a brief incubation in vitro in normal human serum . The low level of CMOA in the peritoneal fluid paralleled a depletion of complement in the peripheral blood . Thus, in cases complicated by sepsis with gram-negative bacteria, the CMOA in the blood was extremely low . These data suggest that the high susceptibility of the peritoneum to infection by gut flora, despite the normal phagocytic response, may be partly explained by the low local level of functional complement which is inadequate for optimal opsonization of the bacteria.

Circ Shock, 1985, 15(1), 5 - 14
Efficacy of S-2441, a synthetic oligopeptide, in a rat model for gram-negative bacteremia; Balis JU et al.; In vitro effects of S-2441, H-D-Pro-Phe-Arg-NH-Heptyl, include potent anti-bradykinin activity and broad-spectrum inhibition of serine proteases involved in the coagulation cascade . In this study, rats infused with 7.8 X 10(8) viable Escherichia coli were treated either with saline (group A) or with intravenous (0.1 mg) and intraperitoneal (0.4 mg) doses of S-2441 (group B) . Survival rates for groups A and B were 68% and 98%, at 12 hours (P less than 0.001), and 37% and 73% at 24 hours (P less than 0.001), respectively . Hematologic studies revealed that S-2441 significantly inhibited E . coli-induced prolongation of prothrombin time and partial thromboplastin time as well as a rapid decrease in the values of factor X, anti-thrombin III, and fibrinogen . In addition, S-2441 attenuated E . coli-induced hypoglycemia and a marked reduction of serum complement level . Ultrastructural evaluation of the liver demonstrated that S-2441 prevented the development of extensive sinusosoidal microthrombosis and hepatocellular necrosis . The results indicate that S-2441 affords protection in lethal gram-negative bacteremia owing in part to attenuation of disseminated intravascular coagulation and complement-mediated reactions . The findings are consistent with the concept that S-2441 and related oligopeptides modulate serine protease-mediated responses involving inhibition of active enzymes with competitive antagonism of pharmcologically active products formed during the activation of coagulation, fibrinolytic, kallikrein, and complement systems.

J Antimicrob Chemother, 1985 Jan, 15(1), 119 - 21
Penetration of ceftazidime into the human prostate gland following intravenous injection; Abbas AM et al.; The concentration of ceftazidime in human prostatic tissue obtained by transurethral resection was measured by microbiological assay in 24 patients after the intravenous administration of one 2 g dose . The average concentration of ceftazidime in the tissue was 10.1 micrograms/g 70 min after injection and 6.1 micrograms/g 140 min after injection; these levels were each approximately 14% of the corresponding serum levels . The concentration of ceftazidime achieved in the prostatic tissue was greater than the minimum inhibitory concentrations of ceftazidime for Gram-negative pathogens.

J Emerg Med, 1985, 3(6), 475 - 83
Septic shock (Part 1); Berringer R et al.; Septic shock is a distinct clinical entity with an overall mortality of 30% to 40% . Gram-negative organisms are the most frequently identified causal agents . The shock syndrome appears to result from activation of complement, coagulation, kinin, endorphin, and other hormonal systems . Physiologic abnormalities include hypotension, metabolic acidosis, increased cardiac index, and decreased tissue extraction of oxygen . Clinicians must entertain the diagnosis of septic shock in a variety of settings, as prognosis is affected by early institution of therapy . Treatment is directed toward control of infection with antibiotics and surgical drainage and cardiorespiratory support utilizing fluids and pressor agents . Future therapeutic options may include naloxone, cyclooxygenase inhibitors, and antiserum to endotoxin.

J Emerg Med, 1985, 3(5), 387 - 94
Rapid nighttime evacuation of a veterans hospital; Blaser MJ et al.; Loss of essential utilities and danger of explosion forced a rapid nighttime winter evacuation of 229 patients from an acute-care Veterans Administration hospital . Although distribution of patients to recipient hospitals was not optimal, and the location of several patients could not be documented for more than 24 hours, the evacuation in subfreezing weather went smoothly . Continuity of care and careful planning permitted an orderly return to the hospital five days later . Although financial costs were high, no excess mortality or morbidity was associated with the evacuation . No changes in pharyngeal gram-negative bacterial flora of the patients were noted . Further, a critique is presented to aid in planning for similar emergencies elsewhere.

Plasmid, 1985 Jan, 13(1), 70 - 4
A small mobilizable IncP group plasmid vector packageable into bacteriophage lambda capsids in vitro; Selvaraj G et al.; A mobilizable cosmid derivative of an IncP group plasmid was constructed by cloning the oriT region of RK2, a wide host-range plasmid, and the minimal DNA sequence of bacteriophage lambda required for efficient packaging in vitro . This cosmid is 13 kb in size and has unique restriction sites for EcoRI, XhoI, HindIII, and SalI . The XhoI and HindIII sites are within the kanamycin-resistance gene and the SalI site is in the tetracycline-resistance gene . This plasmid was mobilizable from an Escherichia coli donor to a number of diverse gram-negative bacteria at a frequency of 0.8 to 10 per 100 donors . This vector is one of the smallest of all wide host-range cosmids described in the literature . As part of this study, another mobilizable IncP group plasmid vector has also been constructed which, in addition to the sites listed above, has a unique BglII site, but which lacks the packager sequence.

Am J Med, 1984 Dec 21, 77(6A), 43 - 50
Role for dual individualization with cefmenoxime; Schentag JJ et al.; Cefmenoxime concentration/effect relationships were retrospectively explored for gram-negative bacteria isolated from 14 critical care patients treated for nosocomial pneumonia . The effects of cefmenoxime concentrations on in vitro growth kinetics of 21 isolated pathogens were studied using the Abbott MS-2 Research System, from which a dynamic response concentration was derived . Serum pharmacokinetic profiles were obtained in each patient . These data were used to calculate the in vivo total area under the curve over dynamic response concentration and the time that cefmenoxime concentrations exceeded the dynamic response concentration for each bacteria . The same determinations were made in 18 patients prospectively treated, except that dosage was optimized on the basis of previous mathematical relations to achieve bacterial eradication in four days . This method of dosage optimization is termed dual individualization . Serial cultures of infected tissues were evaluated to determine the number of days to the eradication of bacteria, and the pharmacokinetic and pharmacodynamic variables were used to describe the bacteriologic response of the original pathogen isolated in pretreatment culture . Bacterial eradication rates could be described from cefmenoxime pharmacokinetics in the patient and from the relation between concentration and bacterial inhibition . Patients who were prospectively treated using these retrospectively derived relationships had a predictable day of bacterial eradication . This, in turn, was associated with a shorter duration of treatment (p less than 0.05) . The success of prospective dual individualization is encouraging and suggests that more precise optimization of antibiotic dosage can yield a predictable rate of bacterial eradication from the infection site.

Infect Control, 1984 Dec, 5(12), 559 - 61
Peritonitis from automated peritoneal dialysis; Hamory BH et al.; Three consecutive patients undergoing automated peritoneal dialysis developed peritonitis with a nonfermenting gram-negative rod . The same organism was recovered from the dialysate produced by the machine and from its tap water supply . This is the first such outbreak in which the causative organism was also recovered from the tap water supply to the machine . The organism persisted in the machine following standard 3% formaldehyde disinfection, but was eradicated by 10% formaldehyde disinfection.

J Lab Clin Med, 1984 Dec, 104(6), 873 - 81
Elevated plasma phospholipase A2 levels: correlation with the hemodynamic and pulmonary changes in gram-negative septic shock; Vadas P; High levels of circulating phospholipase A2 (PLA2) have been implicated in the pathogenesis of hypotension in experimental endotoxin shock . In an attempt to detect similar mechanisms in clinical gram-negative septic shock, plasma samples from 34 patients with hypotension and septicemia were collected and assayed for PLA2 activity . PLA2 levels in 100% of patients with septic shock were elevated (as high as 43-fold over control levels), and mean plasma PLA2 activity was increased 16-fold over control levels . Those patients with concurrent adult respiratory distress syndrome had a mean plasma PLA2 activity level 20.2-fold greater than did controls and 44% greater than patients whose clinical courses were not complicated by adult respiratory distress syndrome . In patients who survived the hypotensive episode, follow-up determinations during the convalescent phase showed a return in plasma PLA2 levels to baseline values . Using pooled plasma, preliminary molecular weight determination revealed the PLA2 to be a 14,000-dalton lipolytic enzyme with 2-acyl specificity . Our study composes the first demonstration of markedly elevated levels of plasma PLA2 during septic shock . Because circulating PLA2 had been shown experimentally to cause profound systemic hypotension and degradation of pulmonary surfactant, this suggests a mechanistic relationship between the massive release of PLA2 into the systemic circulation during the course of gram-negative septicemia, and the resultant hypotension and respiratory distress syndrome associated with gram-negative septic shock.

J Bacteriol, 1984 Dec, 160(3), 994 - 1002
Chemical and immunological characterization of lipopolysaccharides from phase I and phase II Coxiella burnetii; Amano K et al.; Lipopolysaccharides (LPSs) isolated from phase I and phase II Coxiella burnetii (LPS I and LPS II, respectively) were analyzed for chemical compositions, molecular heterogeneity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunological properties . The yields of crude phenol-water extracts from phase I cells were roughly three to six times higher than those from phase II cells . Purification of LPSs by ultracentrifugation gave similar yields for both LPS I and LPS II . Purified LPS I and LPS II contained roughly 0.8 and 0.6% protein, respectively . The fatty acid constituents of the LPSs were different in composition and content, with branched-chain fatty acids representing about 15% of the total . beta-Hydroxymyristic acid was not detected in either LPS I or LPS II . A thiobarbituric acid-periodate-positive compound was evident in the LPSs; however, this component was not identified as 3-deoxy-D-mannooctulosonic acid by gas and paper chromatographies . LPS II contained D-mannose, D-glucose, D-glyceromannoheptose, glucosamine, ethanolamine, 3-deoxy-D-mannooctulosonic acid-like material, phosphate, and fatty acids . LPS I contained the unique disaccharide galactosaminuronyl glucosamine and nine unidentified components in addition to the components of LPS II . The hydrophobic, putative lipid A fraction of LPS I and LPS II contained the above constituents, but the hydrophilic fraction was devoid of ethanolamine . The LPS I disaccharide galactosaminuronyl glucosamine was found in both fractions of the acetic acid hydrolysates . Analysis of LPSs by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by silver staining indicated that LPS II was composed of only one band, whereas LPS I consisted of six or more bands with irregular spacing . Ouchterlony immunodiffusion tests demonstrated that LPS I reacted with phase I but not with phase II whole-cell hyperimmune antibody, and LPS II reacted neither with phase I nor phase II hyperimmune antibody . From these results, it was concluded that the chemical structures of LPSs from C . burnetii were different from those of the LPSs of gram-negative bacteria; however, the LPS structural variation in C . burnetii may be similar to the smooth-to-rough mutational variation of saccharide chain length in gram-negative bacteria.

Am J Med Sci, 1984 Dec, 288(5), 200 - 7
The value of serial serum angiotensin converting enzyme determinations in hospitalized patients with lung disease; Siefkin AD et al.; Abnormal serum angiotensin converting enzyme (ACE) activity has been reported in various human lung disorders and in laboratory animals with acute lung injuries . To test the value of serum ACE activity as an indicator of lung damage and its assistance in diagnosis or prognosis, 328 serum samples were obtained from 108 hospitalized patients with lung disease and 26 normal subjects . When patients were clinically grouped by disease entity, only the sarcoidosis group showed elevated mean serum ACE . Significantly increased serum ACE was found in 17 patients with various lung diseases (15% of hospitalized patients) 12 of whom also had concomitant liver disease . It is hypothesized that the liver may play a role in the normal metabolism of ACE being released by lung endothelial injury . Significantly low levels were seen in many acute and chronic lung injuries; specifically the groups with chronic obstructive lung disease, lung cancer, acute pneumonia, aspiration pneumonitis, gram-negative sepsis, acute myocardial infarction, and congestive heart failure . Serial measures of ACE in 71 patients with lung injuries showed that significantly decreasing levels over successive days were associated with a very high mortality . A single ACE measurement did not predict the presence or extent of lung injury, or aid in diagnosis or prognosis, but serial levels are of value prognostically.

J Clin Microbiol, 1984 Dec, 20(6), 1190 - 1
Recognition of Pneumocystis carinii by gram stain in impression smears of lung tissue; Felegie TP et al.; In 12 of 20 (60%) biopsy-proven cases of Pneumocystis carinii pneumonia, the diagnosis was first suggested by examination of routine Gram stains of impression smears made from infected lung tissue and later confirmed by methenamine-silver staining . The cysts appeared as 5- to 7-microns unstained spheres, each containing six to eight intracystic gram-negative bodies (sporozoites) . Although the Gram stain does not appear to be as sensitive as more traditional staining techniques for the detection of P . carinii, clinical microbiologists should be aware of the morphology of this organism in gram-stained specimens because this relatively simple procedure gives quick results.

Antimicrob Agents Chemother, 1984 Nov, 26(5), 730 - 3
Penetration of aztreonam into cerebrospinal fluid of patients with and without inflamed meninges; Duma RJ et al.; Aztreonam was administered as a single, 2-g intravenous dose to 25 patients with noninflamed meninges and to 9 patients with inflamed meninges . It was well tolerated and was detected in the cerebrospinal fluid at the initial sampling period at 1 h after the end of infusion . Aztreonam levels in the cerebrospinal fluid of patients with inflamed meninges were four times higher than those recorded for the same time period in patients with noninflamed meninges . Aztreonam concentrations in cerebrospinal fluid in the presence of normal and inflamed meninges exceeded the inhibitory and bactericidal concentrations for most gram-negative bacteria . Thus, a multiple-dose treatment regimen with 2-g intravenous doses every 6 h appears to be appropriate for clinical trials of aztreonam for the treatment of gram-negative bacillary meningitis which is caused by susceptible organisms.

Behring Inst Mitt, 1984 Nov, (76), 59 - 74
Antibody-independent killing of gram-negative bacteria via the classical pathway; Clas F et al.; It has been recognised since 1895 that some gram-negative bacteria are sensitive towards the lytic action of serum . Many aspects of this phenomenon in regard to antibody-dependent activation of the complement system and the activation of the alternative pathway in the presence and absence of antibodies had been investigated . However, a lot of serum-sensitive bacteria are killed in nonimmune sera and bind directly C1 in the absence of antibodies . Therefore, we were interested in the killing capacity of an antibody-independent activated classical pathway . For the immediate killing of these serum-sensitive bacteria within even one hour, all complement components are essential . The effective bactericidal effect is dependent on the classical pathway components like C1, C4, C2 and Ca2+ . C1 is directly bound to the bacteria, becomes activated and is able to cleave C4 . For C2-conversion and the further activation of the cascade, an additional serum factor different from an antibody is required . This factor seems to mediate the attachment of C4b to the bacterial surface, which is a prerequisite for the formation of the classical C3-convertase, C4b2a, on the cell surface . The antibody-independent interaction with C1 occurs via C1q, which binds to LPS and possibly also via another C1-subcomponent, C1r and/or C1s . The latter is supposed to interact with outer membrane proteins providing the tight interaction of C1 with the bacteria . This mechanism might be of importance for the killing of R-forms of gram-negative bacteria.

J Mol Cell Cardiol, 1984 Nov, 16(11), 1075 - 9
Neutrophil-derived, oxygen free radical-mediated cardiovascular dysfunction; Rowe GT et al.; Oxygen free radicals and their metabolites generated from activated neutrophils have been implicated in mediating the cardiovascular dysfunction of such diverse etiologies as myocardial ischemia and reperfusion injury, Gram negative sepsis, myocarditis and acute cardiac allograft rejection, but a direct demonstration of neutrophil derived oxygen free radical mediation of cardiovascular dysfunction has not been accomplished . In this study, we have demonstrated that activation of the canine neutrophil system, in vivo, results in the generation of oxygen free radicals that are capable of disrupting cardiovascular function producing a significant decrease in mean arterial pressure and cardiac index without any significant effect on the conduction system of the myocardium . Neutrophil depletion or pretreatment with superoxide dismutase and catalase inhibited the effects of activated neutrophils . This study provides evidence that neutrophil-derived reduced oxygen intermediates are able to induce severe cardiovascular dysfunction.

J Pediatr, 1984 Nov, 105(5), 726 - 30
Peritonitis with continuous ambulatory peritoneal dialysis and continuous cycling peritoneal dialysis; Warady BA et al.; The frequency and cause of peritonitis in 18 children receiving continuous ambulatory peritoneal dialysis (CAPD) and nine children receiving continuous cycling peritoneal dialysis (CCPD) are described . Cumulative CAPD and CCPD experience demonstrated 58 episodes of peritonitis in 294 patient treatment months (one case per 5.1 patient treatment months) . Total hospitalization for the treatment of peritonitis was 0.18 days per patient treatment month . Life table analysis revealed no significant difference in the peritonitis-free "survival" between the two modalities . Gram-negative organisms accounted for a significantly increased percentage of the peritonitis in CAPD compared with CCPD (65% vs 17%) (P less than 0.001) . Thirty-seven percent of the gram-negative infections in the CAPD population were in children with nephrostomies . Factors predisposing to peritonitis were identified in 76% of cases occurring with CAPD . Peritonitis remains the major contributor to the morbidity associated with peritoneal dialysis, regardless of the technique . The resultant frequency of hospitalization is not prohibitive . Attention to the "high-risk" pediatric patient and education directed at several well-recognized predisposing factors may yield improved results.

J Clin Pathol, 1984 Nov, 37(11), 1282 - 8
Postmortem radiographic, histological and bacteriological studies of terminal respiratory infections and other pulmonary lesions in hospital and non-hospital necropsies; Paakko P et al.; One lung from 109 unselected hospital and 43 unselected non-hospital necropsies was studied using postmortem radiography with histological verification . Signs of acute bacterial infection, including bronchiolitis or bronchopneumonia, were present in 42.2% of the hospital necropsies and infection was the immediate cause of death in 8.3% . There was only one case of incipient bronchopneumonia among the non-hospital necropsies . Emphysema was the most common other pulmonary lesion in both groups . Gram negative bacteria were found significantly more often in the bronchial mucus of lungs with any pulmonary lesion, particularly those with signs of bacterial infection, than in the bronchial mucus of morphologically normal lungs in both groups of patients . In addition, the frequency of infections was associated with length of stay in hospital . This may reflect the prevalence of Gram negative infections in hospitals.

Infect Immun, 1984 Nov, 46(2), 501 - 6
Lipopolysaccharide hyperreactivity of animals infected with Trypanosoma lewisi or Trypanosoma musculi; Ferrante A et al.; Rats and mice infected with Trypanosoma lewisi and Trypanosoma musculi, respectively, showed hyperreactivity to lipopolysaccharide (LPS) from gram-negative bacteria . Fatal shock could be precipitated with a dose of LPS 100 to 1,000 times less in infected compared with noninfected animals . In trypanosome-infected rats and mice, extensive liver damage was evident after LPS challenge . These animals showed a pronounced hypoglycemia, marked elevation of blood aspartate transaminase level, and diffuse severe degeneration and total depletion of glycogen in hepatocytes . Only minor changes were observed in noninfected animals given the same dose of LPS . No mononuclear phagocytic cell infiltration was observed in the liver of infected animals . The most striking change was the great increase in size and the probable increase in phagocytic activity and number of sinusoidal Kupffer cells . We suggest that elevated Kupffer cell activity in trypanosome-infected animals may play a role in LPS-induced hepatotoxicity.

Br J Ophthalmol, 1984 Nov, 68(11), 828 - 32
Topical immunotherapy for pseudomonas keratitis in rabbits: use of antilipopolysaccharide plasma; Welsh NH et al.; Pseudomonas keratitis is currently treated with antibiotics with a variable success rate . Part of the morbidity caused by pseudomonas is due to the action of lipopolysaccharide (LPS) present on the surface membrane of the bacteria . Specific IgG present in equine anti-LPS hyper-immune plasma has been found to bind to the LPS from a range of Gram-negative bacteria, including pseudomonas, and by activating complement it destroys these bacteria . Anti-LPS plasma was therefore used as a therapeutic agent in experimentally induced pseudomonas keratitis in rabbits . Thirteen out of 15 (86.7%) anti-LPS treated eyes improved, whereas four out of 17 (23.5%) saline treated control eyes improved (chi 2 = 12.76 p less than 0.001) No ill effects were noted when anti-LPS was administered to healthy rabbit or baboon eyes . Anti-LPS thus was protective in pseudomonas keratitis, and clinical trials appear to be warranted.

Clin Orthop, 1984 Nov, (190), 66 - 74
The aminoglycosides; Whelton A; Aminoglycoside antibiotics continue to be indispensable in the management of complex aerobic gram-negative infections . In the United States approximately 4,000,000 patients receive this antibiotic class each year . Although the clinical efficacy of aminoglycosides is unsurpassed, these antibiotics nonetheless have an inherent tendency to produce undesirable side effects . In particular, nephrotoxicity and ototoxicity continue to be the toxicologic side effects that merit special concern in patient management . If careful attention is paid to the dose of an aminoglycoside, the duration of its use, and other clinical issues, such as the state of hydration of the patient, toxic side effects can be reduced to the minimum . Aminoglycosides are chemically and microbiologically ideally suited for inclusion in polymethylmethacrylate bone cements as a delivery system for prevention or treatment of orthopedic surgical infections.

Radiology, 1984 Nov, 153(2), 493 - 4
Hematogenous osteomyelitis with fat-fluid level shown by CT; Rafii M et al.; Computed tomography (CT) revealed fat-fluid (pus) levels in two adult patients who had hematogenous osteomyelitis due to gram-negative organisms: in one patient within an intraosseous cavity, and in another patient within the adjacent suprapatellar bursae . Both patients were hospitalized for other chronic illnesses and had confusing clinical manifestations with non-specific radiographic changes . Observation of fluid levels, in addition to bony erosions, correctly suggested the presence of pus, and in one patient prompted a diagnostic intraosseous aspiration . To our knowledge, this CT sign of hematogenous osteomyelitis has not been previously reported.

Plasmid, 1984 Nov, 12(3), 189 - 96
Plasmid-borne streptothricin resistance in gram-negative bacteria; Tschape H et al.; Plasmids, which code for a new type of drug resistance due to the acetylation of streptothricin and belonging to various incompatibility groups (I1, I2, W3, FII, S, X, and N) were identified in gram-negative bacteria . The gene for the acetyltransferase is closely linked to a gene for the streptomycin/spectinomycin adenylyltransferase AAD-3" on a novel type of a transposon, designated Tn1825 . This is related to Tn7.

J Lab Clin Med, 1984 Nov, 104(5), 693 - 7
Effect of atropine on oral clearance of a radiolabeled sulfur colloid; LaForce FM et al.; Physical clearance is an important oral defense mechanism against gram-negative rods . We describe a simple technique that uses commercially available technetium-99m sulfur colloid to measure oral clearance . Technetium-99m sulfur colloid was sprayed into the mouth, and clearance was measured as the percent decrease in radiolabel counts over 2 hours using a radioisotope camera . Results using this technique compared favorably with clearance data using Tc-99m radiolabeled Escherichia coli . Atropine significantly decreased oral clearance rates of the colloid . Decreased clearance may be an important risk factor in the development of gram-negative rod colonization in hospitalized patients.

Klin Wochenschr, 1984 Oct 15, 62(20), 986 - 91
Endotoxemia in intensive care patients: a longitudinal study with the limulus amebocyte lysate test; Fink PC et al.; In patients with severe underlying disease and in polytraumatized patients, clinical signs of septicemia caused by infections with gram-negative bacteria are observed postoperatively with increasing frequency . Using a photometric LAL test, a longitudinal assessment of LAL reactivity on 41 intensive care patients was performed . Postoperatively, all patients developed a septicemia of different severity with body temperatures greater than 38.5 degrees C . Dividing the individual disease course, related to body temperatures, into three phases (A-C) it was found that independent of the severity of septicemia, the majority of patients (38/41) yielded a positive LAL reactivity . In phase B (body temperature greater than 38.5 degrees C) more plasma samples contained LAL-reactive material than in phase A and C (body temperature less than 38.5 degrees C) . A decline of fever (phase B to C) correlated significantly (P less than 0.05) with the change from positive to negative LAL reactivity . In patients with high leukocyte counts (15-50 X 10(9)/l) a positive LAL reactivity was found more frequently . The majority of patients (21/27) who survived were transferred with negative LAL reactivity to the general wards . The results suggest that single determinations of LAL reactivity are of limited clinical validity . Using the individual profile of LAL reactivity gained through a longitudinal assessment, data upon the development of the disease course can be obtained.

FEBS Lett, 1984 Oct 15, 176(1), 207 - 10
Anti-LPS factor in the horseshoe crab, Tachypleus tridentatus . Its hemolytic activity on the red blood cell sensitized with lipopolysaccharide; Ohashi K et al.; Anti-LPS factor, which inhibits the endotoxin mediated coagulation system in the horseshoe crab, Tachypleus tridentatus, was found to lyse red blood cells sensitized with gram-negative bacterial LPS, but not to lyse unsensitized cells . This hemolysis occurred even at 0 degree C and was completed within 1 min . The binding of anti-LPS factor to LPS must be essential for the hemolysis, because free LPS inhibited the hemolytic action of anti-LPS factor.

J Periodontol, 1984 Oct, 55(10), 567 - 73
Bacterial invasion of periodontal tissues in advanced periodontitis in humans; Manor A et al.; Bacterial invasion of the pocket epithelium and underlying connective tissue was found in seven cases of advanced human periodontitis . Four cases showed invasion of the epithelium as well as the connective tissue while in the other three cases bacterial invasion was limited to the pocket epithelium . The microorganisms observed included cocci, rods, filaments, fusiforms and spirochetes and these were morphologically similar to those observed in the apical zone of the subgingival plaque . Most bacteria showed typical Gram-negative cell walls . Bacteria were seen in enlarged epithelial intercellular spaces and among debris of disintegrated epithelial cells . In the connective tissue the bacteria were seen among remnants of collagen fibers and degenerated fibroblasts . Identification of the invading microorganisms may assist in understanding the pathogenesis of chronic periodontitis.

J Clin Microbiol, 1984 Oct, 20(4), 626 - 9
Pseudomonas mesophilica and an unnamed taxon, clinical isolates of pink-pigmented oxidative bacteria; Gilardi GL et al.; Twenty-one strains of pink-pigmented bacteria, isolated from human clinical specimens and an environmental source, were compared with Pseudomonas mesophilica ATCC 29983 and Protaminobacter ruber ATCC 8457 . These isolates were gram-negative, oxidative rods which were motile by means of a single polar flagellum; gave positive catalase, indophenol oxidase, urease, and amylase reactions; and grew slowly at 30 degrees C . Fourteen isolates conformed to the designated type strains Pseudomonas mesophilica ATCC 29983 and Protaminobacter ruber ATCC 8457 . The remaining seven strains represented an undescribed taxon . These pink bacteria appear to be invaders of debilitated patients with an underlying chronic disease.

Mem Inst Oswaldo Cruz, 1984 Oct-Dec, 79(4), 433 - 7
Some characteristics of the hyperreactivity to bacterial lipopolysaccharide induced in mice by Trypanosoma cruzi infection; Bambirra EA et al.; Mice infected with T . cruzi, Y strain, acquire a high level of susceptibility to the effects of bacterial gram-negative LPS . The LD50 of adult female SW mice to LPS from S . typhosa, decreases from 450 to 2,5 mcg 10-12 days after T . cruzi infection . This hyperreactivity to LPS induced by T . cruzi presents all the characteristics of that found in infection caused by many other agents . During the acute phase of experimental infection with T . cruzi Y strain, mice generally die with a hypovolemic shock very similar to that induced in uninfected animals injected with an adequate dose of bacterial endotoxin . There is evidence for and against the hypothesis that LPS absorbed from the intestinal tract may be involved in the mechanism of death of mice during the acute phase of T . cruzi infection.

Burns Incl Therm Inj, 1984 Oct, 11(1), 65 - 71
Systemic treatment of septicaemia caused by P . aeruginosa with special reference to azlocillin; Shah PM; Of the pathogens causing septicaemia due to Gram-negative bacteria at Zentrum der Inneren Medizin, Frankfurt, over a 7-year period (1974-80), 16.8 per cent were due to P . aeruginosa . Analysis of all septicaemias over this period, however, shows a decrease from 13 per cent in the 3-year period 1974-76 to 8 per cent in 1977-79 and 5 per cent P . aeruginosa septicaemia in 1980 . The overall mortality rate was 66 per cent, most of the patients dying within 24 hours . Azlocillin and Piperacillin are at present the drugs of choice for systemic therapy of pseudomonas infections, preferably in combination with an aminoglycoside.

Am J Med, 1984 Oct, 77(4), 663 - 6
Oral trimethoprim/sulfamethoxazole in attempt to prevent infection after induction chemotherapy for acute leukemia; Henry SA et al.; The efficacy of orally administered trimethoprim/sulfamethoxazole for infection prevention following induction chemotherapy was evaluated in 43 patients with acute leukemia . Twenty patients were randomly assigned to treatment with trimethoprim/sulfamethoxazole during 20 episodes of profound granulocytopenia; 23 patients in the control group were followed through 25 granulocytopenic episodes . The incidences of superficial skin and overall infections were significantly lower in those patients with multiple relapses who received trimethoprim/sulfamethoxazole (p = 0.008); however, there was no difference between the groups in regard to days of fever, days of antibiotic administration, days of hospitalization, or gram-negative rod bacteremia . As a result of this study, this regimen cannot be unequivocally recommended for infection prevention in neutropenic patients with acute leukemia undergoing induction or reinduction chemotherapy.

Int J Pediatr Otorhinolaryngol, 1984 Oct, 8(1), 1 - 11
Experimental background for the use of tympanostomy tubes; Soderberg O et al.; Already within 24 h, splitting of the soft palate in rats caused production of effusion in the attic space . During the next couple of days the effusion material filled the meso- and hypotympanon . The production of a serous fluid in the attic space appeared concomitant with an elevated histamine content in the middle ear cavity, irrespective of whether the ears were ventilated or not . In the artificially ventilated middle ears no effusion appeared in the meso- and hypotympanon and the hypotympanal mucosal lining remained normal . The serous effusion filling the middle ear cavities with the intact tympanic membranes was replaced at about 7 days by a purulent effusion material containing mainly Gram-negative enteric rods . Thus it seems that a serous otitis media and a purulent otitis media occur in a continuum . The change to a purulent infection could be prevented by artificial ventilation, but as soon as the myringotomies were healed or the tympanostomy tubes got clogged a flocculent, pus-like effusion also appeared in these middle ear cavities.

Clin Haematol, 1984 Oct, 13(3), 549 - 55
Immunoprophylaxis and immunotherapy of Gram-negative infections in the immunocompromised host; Glauser MP et al.; Gram-negative infections remain a prominent cause of serious morbidity and mortality in hospitalized patients despite skilful antibiotic therapy and supportive care . A recently developed immunological approach to this problem is based on antiserum to an E . coli mutant (J5) which elicits antibody cross-reactive with a wide range of Gram-negative pathogens . The antitoxic and protective powers of E . coli J5 antiserum have been demonstrated in animal models . In carefully conducted clinical trials, J5 antiserum or J5 plasma of human origin has been established as a potent adjunctive therapy for the severe consequences of Gram-negative bacteraemia and in the prophylaxis of such infections in surgical patients . The question remains open whether such antiserum may have a similar prophylactic power in severely neutropenic patients . Clinical trials currently underway or soon to be started should help to clarify the practical prophylactic power of J5 hyperimmune globulin against shock and death in high-risk patients.

Am J Med, 1984 Oct, 77(4), 657 - 62
Association of aminoglycoside plasma levels with therapeutic outcome in gram-negative pneumonia; Moore RD et al.; To determine the association of aminoglycoside plasma levels with therapeutic outcome in gram-negative pneumonia, the case reports of 37 patients from four prospective, randomized, controlled trials of gentamicin, tobramycin, and amikacin were analyzed . Twenty (54 percent) of these patients had a favorable outcome . Patients with maximal one-hour postinfusion (peak) levels of 7 micrograms/ml or greater for gentamicin and tobramycin or 28 micrograms/ml or greater for amikacin more often had successful outcomes (14 of 20, 70 percent) than those with levels less than this (six of 19, 32 percent) (p less than 0.006) . Patients with overall mean peak levels of 6 micrograms/ml or greater for gentamicin and tobramycin or 24 micrograms/ml or greater for amikacin more often had successful outcomes than those with levels less than this (six of 17, 35 percent) (p less than 0.04) . The initial patient temperature, serum urea nitrogen/creatinine ratio, initial polymorphonuclear leukocyte count, and age were also associated with outcome; but by multivariate analysis, achieving an adequate peak concentration was the most important discriminating factor . These results suggest the potential importance of achieving adequate aminoglycoside levels in patients with gram-negative pneumonia.

Surg Gynecol Obstet, 1984 Oct, 159(4), 319 - 24
The mediated effect of endotoxin and lead upon hepatic metabolism; Kuttner RE et al.; A test was made of the possibility that gram-negative bacterial cell wall lipopolysaccharides acted directly on key glucoregulatory enzymes in rat liver cytosol to cause the characteristic hypoglycemia of severe endotoxemia . Fasted male rats were sensitized to endotoxin by the simultaneous intravenous injection of lead acetate . The minimum systemic dosage of endotoxin necessary to perturb the normal pattern of hepatic glycolytic intermediates was determined by serial testing with diminishing dosages of endotoxin . The hepatocyte concentration of endotoxin was then calculated from this minimum dosage by use of literature data on the fraction of endotoxin delivered to liver cells after a systemic intravenous injection of radiochromium labeled lipopolysaccharides . Accepting a molecular weight of 118,000 daltons for the smallest endotoxin monomer capable of evoking a physiologic response, the molar amount of endotoxin present in 1 gram of hepatocytes was readily calculated . The concentration of glucoregulatory enzymes in parenchymal cells was then estimated from other literature sources . It was found that the amount of endotoxin in the hepatocytes was insufficient to combine directly with even 1 per cent of the quantity of a single key glucoregulatory enzyme in liver parenchyma . Since a one to one stoichiometric reaction between endotoxin and enzyme could not occur in the liver cytosol, a direct interaction mechanism between agonist and biocatalyst can be ruled out . It is concluded that bacterial endotoxin must act on hepatic glucoregulation by an indirect mechanism presumably based upon the release and operation of mediators.

J Bacteriol, 1984 Oct, 160(1), 299 - 303
Cell wall receptor for bacteriophage Mu G(+); Sandulache R et al.; The invertible G segment in phage Mu DNA controls the host range of the phage . Depending on the orientation of the G segment, two types of phage particles, G(+) and G(-), are produced which recognize different cell surface receptors . The receptor for Mu G(+) was located in the lipopolysaccharide (LPS) of gram-negative bacteria . The analysis of different LPS core types and of mutants that were made resistant to Mu G(+) shows that the primary receptor site on Escherichia coli K-12 lies in the GlcNAc beta 1 . . . 6Glc alpha 1-2Glc alpha 1-part at the outer end of the LPS . Mu shares this receptor site in E . coli K-12 with the unrelated single-stranded DNA phage St-1 . Phage D108, which is related to Mu, and phages P1 and P7, which are unrelated to Mu but contain a homologous invertible DNA segment, have different receptor requirements . Since they also bind to terminal glucose in a different configuration, they adsorb to and infect E . coli K-12 strains with an incomplete LPS core.

Am Fam Physician, 1984 Oct, 30(4), 169 - 73
Branhamella catarrhalis: an unappreciated pulmonary pathogen; Roth RM et al.; Branhamella catarrhalis is being increasingly identified as an important pulmonary pathogen, particularly in elderly patients with chronic obstructive lung disease . This gram-negative diplococcus can readily be isolated and detected in respiratory secretions . Laboratory identification requires care and beta-lactamase testing . Recognition of lower respiratory tract disease caused by B . catarrhalis is rewarding, since safe and effective therapy exists to treat these patients.

J Antimicrob Chemother, 1984 Oct, 14(4), 431 - 4
Penetration of cefotaxime into heart valves, subcutaneous and muscle tissue of patients undergoing open-heart surgery; Just HM et al.; Twenty-nine adult patients undergoing open-heart surgery were given 2 g cefotaxime as 5 min intravenous bolus injection preoperatively . Within 6 h cefotaxime serum concentrations declined from 81.0 to 6.3 mg/l . Subcutaneous tissue and muscle concentrations varied between 0.3 and 8.7 micrograms/g, heart valve concentrations varied between 2.2 and 13.0 micrograms/g . Cefotaxime concentrations in heart valves are high enough to inhibit most Gram-negative organisms causing postoperative wound infections and endocarditis.

Kidney Int, 1984 Sep, 26(3), 308 - 18
Effect of hemodialysis and peritoneal dialysis on aztreonam pharmacokinetics; Gerig JS et al.; Aztreonam, a new monobactam, will be widely used because of its broad aerobic gram-negative bacterial coverage and its apparent low risk of allergic phenomena in penicillin/cephalosporin-sensitive patients . We examined aztreonam kinetics in patients during hemodialysis and in the interdialytic period and in patients on continuous ambulatory peritoneal dialysis (CAPD), and related aztreonam to urea clearance (CL) . In hemodialysis patients, aztreonam serum half-life was 7.9 hr between and 2.7 hr during dialysis sessions . CLserum, CLrenal, and CLother were 24.4, 0.5, and 23.9 ml/min, respectively, during the interdialytic period . Four hours of dialysis removed 38.2% (range, 27 to 58%) of antibiotic . CL of aztreonam by hemodialysis was 36.6 to 43.2 ml/min, 50 to 77% greater than interdialytic CL . CL of urea by hemodialysis was 112.4 to 115.6 ml/min; CLaztreonam/CLurea ratio was 0.28 to 0.33 during the hemodialysis sessions . During CAPD, aztreonam serum half-life after intravenous dosing was 7.1 hr; dialysate recovery, 9.7% of the dose; CLserum, CLrenal, CLperitoneal dialysis, and CLother were 23.8, 0.5, 2.1, and 21.3 ml/min, respectively . CLurea by CAPD was 6.5 ml/min . Thus, CLaztreonam during CAPD was 32% of CLurea . Aztreonam was detectable in dialysate at 48 hr (eight exchanges) after peritoneal administration in the first exchange . Hemodialysis and CAPD patients given aztreonam treatment should receive the standard dose of aztreonam as a loading dose, followed by one-fourth the loading dose at standard dose intervals . Hemodialysis patients should receive a supplemental dose equal to half their usual maintenance dose immediately after each dialysis session . For CAPD patients with peritonitis due to susceptible organisms, a 1-g i.v . loading dose followed by a 0.5-g i.p . dose every 6 hr is suggested . In any individual patient undergoing hemodialysis or CAPD, the relationship between CLurea and CLaztreonam should allow appropriate antibiotic dose adjustment.

J Clin Microbiol, 1984 Sep, 20(3), 579 - 81
Endocarditis caused by Actinobacillus actinomycetemcomitans; Schack SH et al.; Actinobacillus actinomycetemcomitans is a gram-negative coccobacillus which is a very rare cause of bacterial endocarditis . Preexisting cardiac lesions are a main contributing factor, and antibiotic prophylaxis has long been felt necessary before dental or other manipulation to prevent endocarditis . Penicillin in combination with an aminoglycoside has been the most often used treatment regimen . We present a case of endocarditis caused by this organism which developed after antibiotic prophylaxis for dental cleaning . Streptomycin and rifampin therapy resulted in the cure of the infection . The treatment and epidemiology of Actinobacillus endocarditis are reviewed.

Fed Proc, 1984 Sep, 43(12), 2755 - 9
Regulation of the host response to bacterial lipopolysaccharides; Ulevitch RJ et al.; Bacterial endotoxins or lipopolysaccharides (LPS) are unique glycolipids present in the outer cell membrane of all gram-negative bacteria . It is now generally recognized that LPS is of primary importance in initiating the pathophysiological changes that often accompany gram-negative bacillary infections in humans including hypotensive shock, disseminated intravascular coagulation, and metabolic abnormalities . Although the biochemical mechanisms of these changes are not well understood, increasing emphasis has been placed on defining the biochemical response of the macrophage (M phi) to LPS . In this paper we describe two M phi-derived factors induced by LPS that may be important in the expression of endotoxic activity in the host . These are a procoagulant activity, which is present on the cell membrane of LPS-treated rabbit liver M phi and acts by directly activating coagulation factor X, and a factor released into the supernatant by LPS-treated peritoneal exudate M phi, which suppresses steroidogenesis in explanted adrenocortical cells . The potential role of the M phi in regulating the binding of LPS to high-density lipoproteins through the induction of acute phase proteins is also considered.

J Infect Dis, 1984 Sep, 150(3), 380 - 8
Kinetics of endotoxin release during antibiotic therapy for experimental gram-negative bacterial sepsis; Shenep JL et al.; For study of the kinetics of endotoxin release from bacterial cells during therapy for gram-negative bacterial sepsis, serial blood samples were obtained from rabbits with Escherichia coli sepsis that were treated with either antibiotic or placebo . The concentrations of viable bacteria, free endotoxin, and total endotoxin in each blood sample were quantitated . In animals treated with placebo the concentration of free endotoxin was proportional to the level of bacteremia . In contrast, in animals treated with antibiotic the plasma levels of free endotoxin increased 10- to 2,000-fold, in spite of decreasing levels of bacteremia . Free endotoxin that was present in the plasma following antibiotic treatment appeared to be derived in part from the breakdown of circulating bacteria and in part from the disintegration of bacteria in tissues other than the blood . The results of this study demonstrate that significant amounts of endotoxin are released from bacterial cells following administration of antibiotics in vivo.

Arch Surg, 1984 Sep, 119(9), 1021 - 4
Steroid effect on capillary permeability in gram-negative septic shock . Evaluation by vitreous fluorophotometry; Tom WW et al.; The purpose of this study was to use vitreous fluorophotometry to evaluate the effects of glucocorticoids on capillary permeability during gram-negative septic shock . Twelve preconditioned mongrel dogs were cannulated with arterial lines and Swan-Ganz catheters for complete hemodynamic monitoring . All 12 dogs were injected with fluorescein and live Escherichia coli intravenously . Six of the 12 dogs were administered methylprednisolone at set intervals after the initiation of the E coli infusion . Vitreous fluorophotometric recordings were taken hourly . Leakage of fluorescein into the extravascular space occurred in both groups, up to several hours after E coli infusion . However, in the steroid group, the capillary permeability decreased and stabilized following the initial increase in permeability . We conclude that steroids, when administered during the development of gram-negative septic shock, may modify capillary permeability changes.

Invest Ophthalmol Vis Sci, 1984 Sep, 25(9), 1074 - 9
The inflammatory role of endotoxin in rabbit gram-negative bacterial endophthalmitis; Jacobs DR et al.; The authors used the limulus lysate assays to measure the amount of gram-negative endotoxin produced in two rabbits with experimentally induced gram-negative (Escherichia coli) endophthalmitis . A similar amount of purified enodotoxin was injected into the eyes of 14 rabbits to determine the rate of clearance of endotoxin from the rabbit eyes . Endotoxin was found in clinically inflammatory quantities 2 weeks after injection . Results of pathologic examination showed that endotoxin incites severe inflammatory responses in the eye, affecting the ciliary body, vitreous, choroid, retina, and optic nerve . These results suggest that the limulus lysate assay may be useful for detecting early gram-negative endophthalmitis, and that in such cases, early emergency vitrectomy may be needed to remove the inflammation-inciting endotoxin and preserve useful vision.

Antimicrob Agents Chemother, 1984 Sep, 26(3), 358 - 63
Correlation of the effect of beta-lactamase inhibitors on the beta-lactamase in growing cultures of gram-negative bacteria with their effect on the isolated beta-lactamase; Easton CJ et al.; The effectiveness of clavulanic acid, sulbactam, quinacillin sulfone, and the carbapenems MM13902 and MM4550 as inhibitors of TEM-2 beta-lactamase in growing cultures of gram-negative bacteria has been studied . Each of these beta-lactams inhibited the enzyme in intact cells, and the nature of the inhibition correlated with studies on the purified enzyme . The potency of these compounds as inhibitors of the beta-lactamase in vivo can be correlated with the amounts hydrolyzed by the purified enzyme under saturating conditions during the inhibition of the enzyme in vitro.

J Antimicrob Chemother, 1984 Sep, 14 Suppl B, 203 - 11
The prevention of superinfection in multiple trauma patients; Stoutenbeek CP et al.; In a control group of 59 multiple trauma patients requiring prolonged intensive care, a conventional restrictive antibiotic policy was followed . Forty-eight patients (81%) developed 94 infections . Fifty-one patients received systemic antibiotic therapy with one or more drugs . The total quantity of systemic antibiotics used was very high (18.3 +/- 22.1 antibiotic days per patient) . This policy resulted in a very high incidence of superinfections (24%) with multiply-resistant Gram-negative bacteria, mostly emerging from the digestive tract (secondary endogenous infections) . Five patients died from infection . A novel technique of infection prevention, based on the maintenance of the colonization resistance and on selective decontamination of the digestive tract in combination with systematic antibiotic prophylaxis with cefotaxime, proved to be very effective . Out of 63 multiple trauma patients, intubated and ventilated for 5 days or more in the ICU, 10 (16%) developed 11 infections . Most infections occurring under this regimen were primary endogenous infections which were treated by continuation of cefotaxime . Only two patients (3%) developed a superinfection with cefotaxime-resistant Gram-negative bacteria . No patient died . This approach to infection prevention in the ICU, shows that prophylactic administration of antibiotics significantly reduced the infection rate of critically ill patients without the development of superinfection.

Ann Intern Med, 1984 Aug, 101(2), 157 - 63
Acute bronchoconstriction induced by cotton dust: dose-related responses to endotoxin and other dust factors; Castellan RM et al.; Fifty-four healthy humans, selected for their acute airway responsiveness to cotton dust, had spirometric tests immediately before and after 6 hours of exposure to card-generated cotton dust from seven different cottons (of several grades and growing regions) . During exposures, we measured airborne concentrations of viable fungi and bacteria (total and gram negative), vertically elutriated gravimetric dust, and vertically elutriated endotoxin . Correlation between each of these five exposure indices and exposure-related acute changes in forced expiratory volume in 1 s showed a statistically significant relationship between all of the indices except concentration of viable fungi . Of the other four indices, endotoxin was the most highly correlated (r = -0.94; p less than 0.00001), and gravimetric dust was the least correlated (r = -0.34; p less than 0.05) . These findings suggest that gram-negative endotoxin may play a major role in the acute pulmonary response to inhaled cotton dust.

Antibiotiki, 1984 Aug, 29(8), 580 - 5
{Electron microscopic characteristics of the action of penicillin and vancomycin on Rickettsia conorii and Rickettsia akari in vitro}; Barkhatova OI et al.; The possibility of L-transformation for two species of Rickettsia of the tick group (R . conorii and R . akari) was studied with light and electron microscopy . The experiments were performed on the cultures of L-cells (murine fibroblasts) . Penicillin and vancomycin were used as the transforming agents . Electron microscopy revealed clear changes in the morphological structure of both species when the doses of penicillin and vancomycin were 50-100 and 250-1500 micrograms/ml, respectively . The antibiotics had a morphologically analogous effect on both species of Rickettsia . The resulting morphological structures were large oval spheroplast-like cells with significant expansion of the periplasmic space up to 1 X 1.75 micron under the effect of penicillin and 0.5 X 0.7 micron under the effect of vancomycin . R . akari was somewhat more sensitive to the antibiotics . Electron microscopy of the morphological changes in the Rickettsia induced by penicillin and vancomycin showed that they were similar to the changes observed in gram-negative bacteria on their L-transformation and to the penicillin forms of chlamydia (galprovia) and R . prowazekii . They may be considered as a phase of unbalanced growth and represent the initial stage of L-transformation.

J Trauma, 1984 Aug, 24(8), 742 - 6
Problems in the management of type III (severe) open fractures: a new classification of type III open fractures; Gustilo RB et al.; Between 1976-1979, 87 Type III open fractures (in 75 patients) were treated at the Hennepin County Medical Center . Factors leading to increased morbidity in Type III fractures were: massive soft-tissue damage; compromised vascularity; severe wound contamination; and marked fracture instability . This study demonstrates, because of varied severity and prognosis, that the current designation of Type III open fracture is too inclusive . We recommend, therefore, that Type III open fractures be divided, in order of worsening prognosis, into three subtypes . Type IIIA--Adequate soft-tissue coverage of a fractured bone despite extensive soft-tissue laceration or flaps, or high-energy trauma irrespective of the size of the wound . Type IIIB--Extensive soft-tissue injury loss with periosteal stripping and bone exposure . This is usually associated with massive contamination . Type IIIC--Open fracture associated with arterial injury requiring repair . Wound sepsis in the three subtypes were: Type IIIA, 4%, IIIB, 52%; and IIIC, 42%; while amputation rates were, respectively, 0%, 16%, and 42% . Only two patients developed osteomyelitis, and 12 patients had delayed or nonunions . Five patients died, all as a result of multisystem trauma . The bacterial pathogens in infected open fractures have changed dramatically over the years . In the present series (1976-1979), 77% of infections were due to Gram-negative bacteria, compared with 24% previously (1961-1975) . A change of antibiotic therapy from a first-generation cephalosporin alone to a combination of a cephalosporin and an aminoglycoside, or a third-generation cephalosporin, is currently indicated in Type III open fractures.

Eur J Biochem, 1984 Aug 1, 142(3), 541 - 9
The composition of the pyruvate dehydrogenase complex from Azotobacter vinelandii . Does a unifying model exist for the complexes from gram-negative bacteria?
Bosma HJ, de Kok A, Westphal AH, Veeger C.
An improved purification procedure of the pyruvate dehydrogenase complex of Azotobacter vinelandii is described . This procedure minimizes losses of components and results in the isolation of the pure complex with a specific activity of 15-19 U/mg and an overall yield of 40% . The chain ratio of the three components was determined by covalent modification of the lysine residues with trinitrobenzene sulfonic acid, followed by separation of the components on sodium dodecyl sulfate gels . These determinations yielded an average chain ratio of 1.3:1:0.5 for E1:E2:E3 respectively . Based on E2 this corresponds with a minimum molecular mass of approximately 216 kDa . Because the molecular mass of the complex has been determined previously to be 800 +/- 50 kDa, it is concluded that the complex as isolated from A . vinelandii is based on a tetramer of E2 chains . The complex can be resolved into its individual components, which can be recombined to yield a fully active complex . Titration of E2E3 subcomplexes with E1 resulted in maximum complex activity at an E1/E2 ratio of 1.5-1.6 . Similar titrations of E1E2 subcomplexes with E3 resulted in maximum activity at an E3/E2 ratio of 0.45-0.55 . From these experiments it is concluded that the complex has maximum activity with a composition of three E1 dimers, one E2 tetramer and one E3 dimer . With excess of either E1 or E3 a decrease in activity is observed which indicates competition between these components for binding sites on E2 . As shown before {Bosma, H.J., de Kok, A., Markwijk, B.W., and Veeger, C . (1984) Eur . J . Biochem . 140, 273-280}, the isolated E2 component is composed of 32 peptide chains of 66 kDa each . Upon addition of E1 or E3, E2 dissociated into tetramers . Dissociation is complete upon the addition of four E1 dimers of four E3 dimers per E2 tetramer . Addition of E1 to saturated E2E3 subcomplex or E3 to saturated E1E2 subcomplex did not result in extra binding but rather in displacement of bound E3 or E1 respectively . It is therefore concluded that the binding sites of E1 and E3 to the E2 chains are either identical or so closely spaced that steric hindrance prevents simultaneous binding of both components . A model is presented based on the cubic structure of the isolated E2 component . In this model the 32 E2 peptide chains are arranged in tetramers in the corners of the cube . This model is discussed in connection with the existing model for the Escherichia coli complex.

Mech Ageing Dev, 1984 Aug, 26(2-3), 165 - 81
Ageing and the immune response--a unifying hypothesis?
Horan MA, Fox RA.
In this article we review the abnormalities in immune function which have been described in relation to ageing . It is suggested that thymic involution may not be the only underlying cause . Many similar changes can be induced by the endotoxins from the cell walls of Gram-negative bacteria . The colon is a large repository for these organism and bacterial breakdown products are found in portal venous blood . Spillover into the peripheral circulation is prevented by the hepatic Kupffer cells degrading these substances . A waning in Kupffer cell function is well documented in association with ageing and there may be spillover of endotoxins into the peripheral circulation in elderly individuals . It is suggested that such spillover of endotoxins may contribute to some of the immunological changes previously attributed to ageing.

Infect Immun, 1984 Aug, 45(2), 531 - 3
Detection of free endotoxin in cerebrospinal fluid by the Limulus lysate test; Munford RS et al.; We used a rabbit model of Escherichia coli meningitis to study the basis for positive Limulus lysate tests in infected cerebrospinal fluid . The results indicated that positive Limulus tests are due to endotoxins in cerebrospinal fluid and not to leukocyte proteases or other possible activators of the Limulus clotting system . The results also suggest that bacteria-free endotoxin may be present in localized gram-negative bacterial infections.

Infect Immun, 1984 Aug, 45(2), 417 - 23
Endogenous interferon production by endotoxin-responsive macrophages provides an autostimulatory differentiation signal; Vogel SN et al.; Previous studies have demonstrated that peritoneal macrophages (resident or thioglycolate-induced) derived from mouse strains fully responsive to gram-negative endotoxins continue to differentiate in vitro, as evidenced by an increased capacity to phagocytose via the Fc receptor with time in culture . In contrast, macrophages derived from endotoxin-hyporesponsive mouse strains (e.g., C3H/HeJ or C57BL/10ScN) exhibit no such increase in phagocytic capacity, and, in fact, significantly lose the capacity to phagocytose particles opsonized with immunoglobulin G with time in culture . This defect was found to be fully correctable by the addition to the cultures of an exogenous source of alpha, beta, or gamma interferon . In this study, we compared C3H/HeN (endotoxin-responsive) and C3H/HeJ (endotoxin-responsive) and C3H/HeJ (endotoxin-hyporesponsive) macrophages in an attempt to elucidate the mechanism responsible for this difference in phagocytic (differentiative) potential . The following observations support the hypothesis that endotoxin-responsive macrophages, in contrast to endotoxin-hyporesponsive macrophages, produce significantly higher levels of an autostimulatory differentiation signal that appears to be macrophage-derived interferon . (i) Anti-alpha/beta-interferon antibody greatly reduces the ability of C3H/HeN macrophages to phagocytose opsonized erythrocytes: (ii) C3H/HeJ macrophages can be made more phagocytic by coculture with C3H/HeN macrophages or by treatment with supernatants derived from C3H/HeN macrophage cultures; and (iii) C3H/HeN macrophages spontaneously lose Mac-1 antigen with time in culture . C3H/HeJ macrophages must be interferon-treated to be equivalently down-regulated.

J Periodontol, 1984 Aug, 55(8), 482 - 5
Histopathological and ultrastructural findings in a case of Papillon-Lefèvre syndrome; Sloan P et al.; Gingiva from a 3-year-old Iranian boy suffering from Papillon-Lefevre syndrome was examined by light and electron microscopy . Deep pockets associated with predominantly Gram-negative plaque were present . The gingival lesion was dominated by plasma cells, many of which were degenerate . Russell bodies were a prominent feature . No defect of epithelium was detected.

J Exp Med, 1984 Aug 1, 160(2), 353 - 68
Virulence of Entamoeba histolytica trophozoites . Effects of bacteria, microaerobic conditions, and metronidazole; Bracha R et al.; The association of axenically grown trophozoites of Entamoeba histolytica strains HK-9 or HM-1:IMSS with various types of gram-negative bacteria for relatively short periods markedly increased their virulence, as evidenced by their ability to destroy monolayers of tissue-cultured cells . Interaction of trophozoites with bacteria that were heat inactivated, glutaraldehyde fixed, or disrupted by sonication, or bacteria treated with inhibitors of protein synthesis, did not augment amebic virulence . Lethally irradiated bacteria, however, retained their stimulative properties and trophozoites that ingested bacteria were protected from the toxic effects of added hydrogen peroxide . An increase in virulent properties of amebae was also found in experiments carried out under microaerobic conditions (5% O2, 10% CO2) . The augmentation of amebic virulence due to association with bacteria was specifically blocked by metronidazole, but not by tetracycline or aminoglycosides, and the rate of metronidazole uptake in stimulated trophozoites was two to three times higher . The results obtained suggest that virulence of axenically grown E . histolytica trophozoites may depend to a considerable extent on the cell's reducing power . Both microaerobic conditions and the association with bacteria apparently stimulate the electron transport system of the ameba . Bacteria may function as broad range scavengers for oxidized molecules and metabolites through the contribution of enzymatic systems, components, or products.

Cell, 1984 Aug, 38(1), 203 - 9
Multicopy single-stranded DNA isolated from a gram-negative bacterium, Myxococcus xanthus; Yee T et al.; A gram-negative bacterium, Myxococcus xanthus, was found to contain 500 to 700 copies per chromosome of a short single-stranded linear DNA fragment . When this DNA (multicopy single-stranded DNA; msDNA) labeled at the 5' end with kinase was used as a probe against total chromosomal blots, it hybridized to unique high molecular weight bands, which were cloned and sequenced . Labeling of msDNA was also possible using the Klenow fragment of DNA polymerase I as well as terminal deoxynucleotidyl transferase, permitting direct sequencing . The 5' end of msDNA was found to be primed by a short RNA segment . The DNA portion of msDNA consisted of 163 bases . Exact correspondence was seen between the msDNA sequence and the sequence of a chromosomal clone . An elaborate secondary structure is postulated for the msDNA sequence . A similar satellite DNA was also found in another myxobacterium, Stigmatella aurantiaca.

J Biol Chem, 1984 Jul 10, 259(13), 8388 - 93
Regulation of the synthesis of membrane-derived oligosaccharides in Escherichia coli . Assay of phosphoglycerol transferase I in vivo; Bohin JP et al.; Membrane-derived oligosaccharides are periplasmic constituents of Escherchia coli and other Gram-negative bacteria . Oligosaccharides in this family may be variously substituted with O-succinyl ester residues, and with sn-1-phosphoglycerol and phosphoethanolamine residues derived from membrane phospholipids . Membrane-derived oligosaccharides appear to be important in osmoregulation, because their synthesis is under strict control (Kennedy, E.P . (1982) Proc . Natl . Acad . Sci . U.S.A . 79, 1092-1095) . Maximum rate of synthesis is at very low osmolarity of the medium . Phosphoglycerol residues are transferred from phosphatidylglycerol to membrane-derived oligosaccharides, or to certain beta-glucoside acceptors, in a reaction catalyzed by phosphoglycerol transferase I, an enzyme of the inner membrane (Jackson, B . J., and Kennedy, E.P . (1983) J . Biol . Chem . 258, 2394-2398) . We now report that this enzyme catalyzes the transfer of phosphoglycerol residues to arbutin (p-hydroxyphenyl-beta-D-glucoside) added to the medium with Km similar to that observed with the cell-free enzyme . The active site of the enzyme must therefore be on the periplasmic face of the inner membrane . We assayed phosphoglycerol transferase I in vivo and found that it is present and completely active even in cells growing in medium of very high osmolarity, in which the synthesis of membrane-derived oligosaccharides is severely reduced . We conclude that osmotic regulation must occur at the stage of the synthesis of oligosaccharide chains . A study of the kinetics of transfer of phosphoglycerol residues to membrane-derived oligosaccharides in vivo revealed that synthesis of the polyglucose chains must stop abruptly upon transfer of cells from medium of low to high osmolarity, inconsistent with a model postulating simple dilution of some rate-limiting enzyme during growth at the higher osmolarity.

J Bacteriol, 1984 Jul, 159(1), 427 - 8
Uptake of D-xylose and D-glucose by Spirochaeta aurantia; Roberts S et al.; Uptake of D-{14C}glucose and D-{14C}xylose by Spirochaeta aurantia was demonstrated to be osmotic shock sensitive and to require a high-energy phosphorylated compound rather than a proton motive force . These features are similar to those of binding protein-mediated transport systems in other gram-negative bacteria.

Equine Vet J, 1984 Jul, 16(4), 376 - 82
Clinical and clinicopathological characteristics of the septicaemic neonatal foal: review of 38 cases; Koterba AM et al.; In an effort to identify improved methods for diagnosis of infection in the neonatal foal, clinical records from 38 septicaemic foals of less than one week of age were examined for trends in history, physical examination and clinicopathological findings . The survival rate of septicaemic foals, 26 per cent, was markedly less than the rate for all other foal admissions . Blood cultures were valuable in diagnosis and treatment of septicaemia and identified a preponderance of Gram-negative infection . Zinc sulphate turbidity test results were abnormally low in all septicaemic foals tested . The clinical course was often distinguished by severe complications and multiple organ dysfunction, leading to death . Conditions present in the mare pre-partum resulted in weak or diseased foals; bacterial placentitis, vaginal discharge and premature lactation were most common . There was no single diagnostic criterion of the septicaemic foal . Fever was not a consistent finding . The most useful white blood cell parameters were neutropenia, the presence of band neutrophils (greater than 0.2 X 10(9)/litre) and toxic changes in the neutrophil population . Hypoglycaemia, metabolic acidosis and hypoxaemia were also common findings.

Postgrad Med J, 1984 Jul, 60(705), 485 - 6
Cholecystitis associated with myelomatosis; Sherlock D et al.; A case of myeloma presenting as acute cholecystitis unresponsive to conventional management is reported . The impairment of the immunological response is a well-known aspect of myeloma, although this usually takes the form of recurrent respiratory infections . It is unusual for acute on chronic cholecystitis, a predominantly Gram-negative infection, to present in this way.

Prikl Biokhim Mikrobiol, 1984 Jul-Aug, 20(4), 435 - 44
{Study methods for localizing enzymes in the cells of gram-negative bacteria (a review)}; Tysiachnaia IV et al.; The present review systematizes the literature data on localization of enzymes in cells of Gram-negative bacteria . It centres round the methods used for studying the enzyme localization . The criteria according to which one can locate an enzyme in bacterial cells are discussed.

Gan To Kagaku Ryoho, 1984 Jul, 11(7), 1356 - 68
{Definition of tumor-necrosis factor and its production mechanism}; Urushizaki I et al.; There is significant evidence that the macrophage plays a critical role in the host's defense against neoplasia . Tumor-necrosis factor was recognized by Carswell et al . during a study of the antitumor activity of serum from mice infected with BCG and subsequently injected with endotoxin . The same procedure was applied to rabbits in order to obtain serum containing tumor-necrosis factor (TNF) . Sera from these mice and rabbits contained a factor that induced hemorrhagic necrosis of certain mouse sarcomas in vivo and had cytotoxic effects on mouse and human tumor cells in vitro . Sera from mice and rabbits singly treated with BCG or endotoxin did not have these properties . Other agents such as C . parvum, OK-432, lentinan or zymosan, that cause hyperplasia of reticuloendothelial system and increase sensitivity to endotoxin lethality, could substitute for BCG in priming for TNF release . However, the use of P . acnes as a priming agent was the most effective and lipopolysaccharide from gram-negative bacteria appeared to be unique in its ability to elicit TNF release . TNF is a protein with a molecular weight, ranging from 40,000 to 60,000 that has both tumor necrotizing activity in vitro and tumor killing activity in vitro . It is relatively stable to heating at up 70 degrees C . This result indicated that both in vitro and in vitro activities of mouse and rabbit TNF are a property of one and the same molecule . TNF is thought to be produced by macrophage and is distinguished from the other know macrophage products in serum containing TNF . TNF is cytotoxic to several but not all tumor cell lines . Its most interesting feature is that it reportedly dose not affect any non-transformed cell types, implying that it somehow recognizes transformed cells.

Am J Med, 1984 Jul, 77(1), 157 - 61
Catecholamine-secreting paraganglioma of the interatrial septum; Hodgson SF et al.; A 29-year-old woman was found to have clinical and biochemical evidence of a functioning paraganglioma (pheochromocytoma) five years after the onset of symptoms . Despite extensive investigation of known potential sites of paragangliomas (including laparatomy), the suspected tumor eluded localization . Adrenergic blocking agents controlled her blood pressure but did not affect the other manifestations of catecholamine excess . Ischemic perforation of the colon and refractory gram-negative sepsis resulted in death . At autopsy, a large paraganglioma was found within the heart; the unique tumor location was an important factor in the fatal clinical course.

J Bacteriol, 1984 Jul, 159(1), 407 - 9
Two tuf genes in the cyanobacterium Spirulina platensis; Tiboni O et al.; Probes derived from the tufA gene of Escherichia coli have been utilized to detect homologous sequences on Spirulina platensis DNA . A 6-kilobase-pair fragment of S . platensis DNA appears to contain two sequences homologous to the E . coli gene . Thus, as reported for gram-negative bacteria, the cyanobacterium presumably contains two tuf genes.

Cell Immunol, 1984 Jul, 86(2), 429 - 38
Suppression of rabbit lymphocyte functions by antibodies specific for allotypic membrane determinants; Ohama Y et al.; Regulation of immunoglobulin synthesis and secretion was analyzed by exposing spleen cells of b4b4 rabbits to anti-b4 for 24 hr in culture . As noted previously, no lymphocytes with membrane-bound b4 were found immediately after pulse treatment, but substantial regeneration of membrane Ig (mIg) occurred on further culture in antibody-free medium . Splenocytes cultured either in the presence or absence of anti-b4 showed a marked loss of Ig-secreting cells (ISC) after 24 hr in culture but recovered and exhibited peak numbers of ISC on Day 2 . However, ISC formation in cultures of antibody-treated cells was significantly suppressed and thereafter declined at a more rapid rate than in control cultures . Polyclonal B cell activators from Nocardia and from gram-negative bacteria stimulated ISC formation in cultures of normal spleen cells, but responsiveness to these activators was depressed following antibody treatment . Antibody-induced suppression of Ig synthesis was attributed to interference with differentiation of B lymphocytes to the secretory stage.

Gene, 1984 Jul-Aug, 29(1-2), 93 - 102
Broad host range cloning vectors for gram-negative bacteria; Sharpe GS; A series of cloning vectors has been constructed based on the broad-host-range plasmid R300B . One of these vectors, pGSS33, has a size of 13.4 kb and carries four antibiotic resistance genes {ampicillin (Apr), chloramphenicol (Cmr), streptomycin (Smr) and tetracycline (Tcr)}, all of which have restriction sites for insertional inactivation . The derivation, structure and uses of the plasmids are described.

Hum Pathol, 1984 Jul, 15(7), 670 - 6
Necropsy findings in acquired immunodeficiency syndrome: a comparison of premortem diagnoses with postmortem findings; Hui AN et al.; Necropsies were performed in 12 patients who fulfilled the Centers for Disease Control (CDC) criteria for acquired immunodeficiency syndrome (AIDS), and the postmortem findings were compared with the premortem diagnoses . All of the patients were men with a male sexual preference and histories of multiple episodes of venereal diseases . Four patients were intravenous drug abusers, while two abused amyl nitrate . All 12 of the patients had evidence of cellular immune deficiency at presentation . The causes of death were a variety of opportunistic infections and neoplasms . Pneumocystis carinii pneumonia was diagnosed prior to death in seven patients . Despite current therapy, all seven of those patients had persistent Pneumocystis carinii pneumonia at necropsy, as well as clinically undiagnosed cytomegalovirus infection . In addition, two cases of acid-fast infections, two of visceral candidiasis, one of pneumocystis pneumonia, one of central nervous system lymphoma, one of gram-negative bacterial pyelonephritis, and one of cutaneous aspergillosis were clinically unrecognized and untreated . Nine patients died with two or more infections . Thus, necropsy is a valuable tool for recognizing clinically undiagnosed infections and malignant disorders in AIDS.

Gastroenterol Clin Biol, 1984 Jun-Jul, 8(6-7), 503 - 6
{Anti-Gram-negative bacterial antibodies in alcoholic cirrhosis . Study of 58 patients}; Bercoff E et al.; Gram-negative bacterial infections are frequent and severe in cirrhotic patients . Existence of endotoxemia in cirrhosis is controversial . The demonstration of Gram-negative bacterial antibodies could be an alternative approach to the pathogenic role of these bacteria . In 58 patients with alcoholic cirrhosis, the immunoglobulin G specifically directed against the Gram-negative bacteria lipopolysaccharide expressed by the J5 mutant of Escherichia coli 0111:B4 was measured . Antibody titres were compared to those of a control group of blood donors . The distributions of antibody titres were similar in cirrhotic patients and in control subjects . No correlation was found between antibody titres and biological parameters of liver function . These results seem to confirm previous reports on the absence of latent endotoxemia in cirrhotic patients, and they suggest that antibody production against Gram-negative bacteria lipopolysaccharides is not enhanced in these patients.

J Clin Invest, 1984 Jun, 73(6), 1772 - 84
Effect of N-acetylcysteine on the pulmonary response to endotoxin in the awake sheep and upon in vitro granulocyte function; Bernard GR et al.; Oxygen free radicals released during endotoxemia may contribute to the lung injury of the adult respiratory distress syndrome (ARDS) . As this syndrome occurs frequently after gram-negative sepsis in humans, we studied the effect of intravenous N-acetylcysteine (NAC), a free radical scavenger, upon the endotoxin (E)-induced model of ARDS in awake sheep . In vivo studies demonstrated that NAC attenuates the endotoxin-induced rise in pulmonary artery pressure (62 +/- 3 torr with E control vs . 43 +/- 3 torr for E + NAC), and markedly diminishes the rise in lymph flow at 1 h (8.5 +/- 1.2 vs 4.5 +/- 0.6 ml/15 min) and 4 h (5.0 +/- 0.6 vs . 3.3 +/- 0.4 ml/15 min), respectively, for E control vs . E + NAC . NAC also markedly attenuated the alterations in lung mechanics after endotoxemia . Dynamic compliance at 2 h after endotoxemia was 44 +/- 6% of base line for E vs . 76 +/- 10% of base line for E + NAC . Resistance to airflow across the lung at 1 h postendotoxin was 811 +/- 280% of base line for E vs . 391 +/- 233% of base line for E + NAC . NAC substantially reduced the 1 h postendotoxin rise in lymph concentrations of thromboxane B2 (8.29 +/- 3.28 vs . 2.75 +/- 1.93 ng/ml for E vs . E + NAC) and 6-keto-prostaglandin-F1 alpha (0.91 +/- 0.27 vs . 0.23 +/- 0.12 ng/ml for E vs . E + NAC) . In addition, in vitro studies were performed which revealed NAC to be a potent free radical scavenger in both biologic and nonbiologic free radical generating systems . NAC decreased phorbol-stimulated granulocyte aggregation in a concentration-dependent manner in vitro . Minimal effects were observed, however, upon leukocyte degranulation at the concentrations of NAC achieved during the in vivo tests . Thus, NAC significantly attenuated all monitored pathophysiologic changes in the endotoxin model of ARDS in sheep, possibly by its ability to scavenge toxic oxygen free radicals . A direct impairment of the ability of inflammatory cells to generate oxygen radicals cannot be ruled out.

Chest, 1984 Jun, 85(6), 729 - 32
Cardiopulmonary function following post-cardiac surgical mediastinitis; Grigas D et al.; Eleven survivors of bacterial mediastinitis, which followed cardiac surgery through a median sternotomy approach, underwent noninvasive cardiopulmonary evaluation to determine whether clinically apparent mediastinal fibrosis had developed . Compared to preoperative values, forced vital capacity was reduced by 9.75 percent of predicted; the greatest reduction occurred in those patients with Gram-negative mediastinitis . In one patient, an abnormal jugular venous wave form and apex cardiogram were consistent with constrictive cardiac physiology, but this was not associated with an abnormal echocardiogram or impaired functional status . In this small series, no serious abnormalities in cardiopulmonary function were detected which could be attributed to prior mediastinal infection.

Arch Latinoam Nutr, 1984 Jun, 34(2), 384 - 90
{Bacterial flora of liver paté in 3 stages of processing}; Gesche E et al.; Liver pate is a boiled sausage that, due to its nature and composition, has an appropriate environment for bacterial development . In order to determine the variation in the bacterial development during the processing of this sausage, bacteriological examinations were carried out in three stages of preparation . This involved, first, the fresh product, then, after boiling it, and finally on the 6th day, after cold storage at 0 degrees C . Of the bacterial counts obtained it was determined that boiling greatly reduces the number of bacteria, especially the Gram negative ones, a situation which lasts through the storage period studied . The relationship between initial and final bacterial content of liver pate, clearly shows the influence of contamination of the raw materials employed, on the microbiological quality of the marketed product.

Antibiotiki, 1984 Jun, 29(6), 403 - 7
{Growth of test microorganisms used in determining antibiotic activity on synthetic nutrient media}; Vasil'eva NG et al.; The growth of gram-negative sporulating bacteria and yeast-like fungi used in a microbilogical assay of antibiotic activity was studied on solid synthetic media of simple composition . Their reproduction with the microbial growth of different density was shown to be possible on media containing available and strictly standardized components, such as salts, glucose and disubstituted sodium phosphate . The cultures tested were not similar by their growth requirements.

EMBO J, 1984 Jun, 3(6), 1315 - 9
Repeating functional domains in the pyruvate dehydrogenase multienzyme complex of Escherichia coli; Packman LC et al.; Each polypeptide chain in the lipoate acetyltransferase (E2) core of the pyruvate dehydrogenase complex from Escherichia coli contains three repeating sequences in the N-terminal half of the molecule . The repeats are highly homologous in primary structure and each includes a lysine residue that is a potential site for lipoylation . We have shown that all three sites are lipoylated, at least in part, and that the three lipoylated segments of the E2 chain can be isolated as distinct functional domains after limited proteolysis . Each domain becomes partly acetylated in the intact complex in the presence of substrate . In the primary structure, the domains are separated by regions of polypeptide chain oddly rich in alanine and proline residues . These regions are probably the conformationally mobile segments observed in the 1H-n.m.r . spectrum of the complex and which are removed by tryptic cleavage at Lys-316 . The C-terminal half of the molecule contains the acetyltransferase active site and the binding sites for E1, E3 and other E2 subunits . The pyruvate dehydrogenase complex of E . coli, which has a heterogeneous quaternary structure, is thus far unique among the 2-oxo acid dehydrogenase complexes in possessing more than one lipoyl domain per E2 chain, but this may be a general feature of the enzyme from Gram-negative organisms.

Dig Dis Sci, 1984 Jun, 29(6), 502 - 7
Circulating lipid A antibodies despite absence of systemic endotoxemia in patients with Crohn's disease; Kruis W et al.; Lipid A is a component common to endotoxins of gram-negative bacteria . It has been suggested that the gut wall of patients with inflammatory bowel disease is more permeable to luminal bacterial macromolecules which may stimulate the gut-associated lymphoid tissue . We therefore investigated 40 patients with Crohn's disease (CD) and 23 patients with ulcerative colitis (UC) with respect to their lipid A antibody titers and presence of endotoxins (Limulus test) . Both tests were performed simultaneously using peripheral venous blood . Systemic endotoxin was demonstrated in only two of the patients . The lipid A antibody titers in the CD patients were significantly higher than either in patients with UC or in 42 healthy controls . Lipid A titers of patients with UC did not differ from those of controls . Titers of lipid A were significantly higher in patients with active Crohn's disease than in patients with inactive disease . It is concluded that systemic endotoxemia occurs rarely in patients with inflammatory bowel disease; however, despite this virtual absence of endotoxins in their peripheral blood, patients with CD show an increase in antibody formation against lipid A . This is suggestive of an altered immunologic reactivity against endotoxins in patients with CD and may be relevant to the pathogenesis of this disease.

J Mol Biol, 1984 May 25, 175(3), 251 - 62
Nucleotide sequence of the trfA gene of broad host-range plasmid RK2; Smith CA et al.; The nucleotide sequence of a 1622 base-pair segment of the broad host-range IncP plasmid RK2 (identical to RP1, RP4, R18 and R68) was determined . This region includes the trfA gene, encoding a trans-acting product essential for vegetative plasmid replication . The nucleotide sequence, together with the results described in the accompanying paper by Shingler & Thomas, indicates that the trfA gene encodes two polypeptide products (of 382 and 285 amino acids) by utilizing different translational start points within a single open reading frame . The region common to both trfA polypeptides includes a sequence with homology to a number of proteins that bind to double-stranded DNA . The trfA gene is preceded by another open reading frame, encoding a polypeptide of 116 amino acids of unknown function . Both cistrons are transcribed from a promoter outside the region of sequence reported here; however, much higher levels of the short polypeptide than of either of the trfA gene products are observed . Possible mechanisms for the control of the relative levels of the products of this operon are discussed, together with features of the trfA gene that may be important for its function in the diverse gram-negative bacterial species in which RK2 can be maintained.

J Mol Biol, 1984 May 25, 175(3), 229 - 49
Analysis of the trfA region of broad host-range plasmid RK2 by transposon mutagenesis and identification of polypeptide products; Shingler V et al.; Broad host-range plasmid RK2 is a member of the Escherichia coli incompatibility group P . Unlike most other groups of plasmids, members of the P group are capable of efficient transfer between and maintenance in most gram-negative bacterial species . It is of interest whether this broad host-range results from differences between the mechanism of replication of broad and narrow host-range plasmids . The regions of RK2 required for replication in E . coli have previously been defined as an origin of vegetative replication, oriVRK2 , and a gene, trfA , specifying a positively required trans-acting product . In this study Tn1723 transposon insertions have been used to map the trfA gene and determine its functional gene product . The Tn1723 insertions define the outer limits of the gene, a promoter region, a "leader" region not essential for trfA activity and a coding region . Three polypeptides of 13 X 10(3), 43 X 10(3) and 32 X 10(3) molecular weight are produced from this region and the production of a 32 X 10(3) Mr polypeptide is shown to be correlated with trfA activity in E . coli . Analysis of polypeptides produced from transposon insertion derivatives in which all but 35 base-pairs of inserted DNA is deleted, along with the effect of these insertions on trfA activity, suggest that the 43 X 10(3) and 32 X 10(3) Mr polypeptide coding sequences overlap in the same reading frame and that all three polypeptides (13 X 10(3), 32 X 10(3) and 43 X 10(3) Mr) may be translated from the same initial transcript.

Am J Med, 1984 May 15, 76(5A), 158 - 65
Infections in burn patients: a paradigm for cutaneous infection in the patient at risk; Gelfand JA; The burn patient is in many ways the archetypical immunosuppressed host: the anatomic barriers have been breached and the host's defenses suppressed . Infection is the leading cause of death in hospitalized burn patients not having inhalation injury, being responsible for between 50 and 75 percent of the hospital deaths . Burn injury produces profound abnormalities in immunologic function . These changes are generally proportional to the degree of burn injury (surface area and depth) . Cell-mediated immune function is suppressed . Anergy and depressed allograft rejection occur . Acute burn serum samples contain several factors that suppress cell-mediated immune functions . Antibody levels are usually mildly to moderately reduced . Complement, particularly the alternative complement pathway, may be massively activated and depleted, removing a critical defense mechanism against gram-negative rod infections for which preformed antibodies are absent . This depression is further exacerbated by malnutrition and infection . Fibronectin levels are also reduced . Phagocytic cell function is abnormal with burns . Neutrophil numbers may be depressed, and function is abnormal . Chemotactic responsiveness, cytoplasmic granule enzyme content, and oxygen radical generation are abnormal . Monocyte/macrophage dysfunction has similarly been demonstrated . Burn infections reflect abnormal host functions as well as changes in the hospital environment and microbial selection pressures . The burn patient is a model of cutaneous infection in the patient at risk.

J Bone Joint Surg Br, 1984 May, 66(3), 340 - 3
A six-year follow-up of infected total hip replacements managed by Girdlestone's arthroplasty; Bourne RB et al.; Thirty-three Girdlestone's arthroplasties used to salvage infected total hip replacements were followed for a mean of 6.2 years to assess the efficacy of this procedure . Subjective and objective assessment was obtained at the time of review . Gram-negative organisms, retained cement, bony sequestra and secondary healing were common features of the five patients (15%) whose wounds discharged for more than four weeks after operation . Girdlestone's arthroplasty provided satisfactory pain relief in 91% of patients, and control of infection in 97% . Although leg-length discrepancy (mean 4 cm) and a Trendelenburg gait made walking difficult and tiring , 42% were satisfied with their functional ability; 85% needed walking aids, and 3 patients could not walk at all and were confined to bed or a chair . Overall, 79% were satisfied . Objectively, the mean Harris hip rating was 60; however, if patients with multiple joint problems were excluded, this score increased to 77 . Girdlestone's arthroplasty seems a reasonable salvage procedure in the management of an infected total hip replacement and the results seem to improve with time.

J Antibiot (Tokyo), 1984 May, 37(5), 577 - 87
Inactivation of cephamycins by various beta-lactamases from gram-negative bacteria; Minami S et al.; The enzymic inactivation of cephamycins, i.e . cefoxitin, cefmetazole, cefotetan and cefbuperazone, was investigated by means of bioassay, high pressure liquid chromatography (HPLC) and spectrophotometric analysis using three types of cephalosporinase (CSase, RICHMOND type Ia, Ib and Ic) and one penicillinase (PCase, TEM type) . These four cephamycins were not inactivated by Ic CSase and TEM type PCase or producers of these enzymes . However, the inactivation of cefmetazole and cefoxitin was noted when they were incubated in the cultures of CSase (Ia and Ib)-producers or incubated with a large amount of these purified enzymes although the inactivation of cefbuperazone was not noted . HPLC of culture fluid or enzyme solution which contained cefmetazole or cefoxitin and were incubated at 37 degrees C showed that metabolites of cefmetazole or cefoxitin appeared as the drug disappeared . In addition, the appearance of metabolites corresponded to the loss of the drug's bioactivities and the absorption of iodine . UV and IR spectra of cefmetazole which were taken after incubation with the purified CSase showed the cleavage of the beta-lactam ring.

Ann Clin Lab Sci, 1984 May-Jun, 14(3), 216 - 24
Role of hydrophobicity in adherence of gram negative bacteria to epithelial cells; Sobel JD et al.; Agglutination of yeast, human group A and guinea pig erythrocytes by multiple clinical isolates of E . coli, K . pneumoniae, and P . aeruginosa was investigated and correlated with hydrophobicity measurements of each bacterial strain . Hydrophobicity of the isolates, as measured by hydrophobic interaction chromatography, was similarly correlated with in vitro adherence of the microorganisms to buccal epithelial cells . Agglutination and adherence studies were done in the presence and absence of 0.046 M D (+) mannose . Results showed a wide variability of these parameters among the three general of bacteria . Although E . coli designated mannose sensitive by agglutination showed significantly greater hydrophobicity and attachment to buccal cells, there was no direct correlation between hydrophobic retention and adherence to epithelial cells (p greater than 0.5) . As a group, K pneumoniae strains adhered in higher numbers than other gram negative species, but this was unrelated to the hydrophobicity or the designated mannose sensitive/mannose resistant adhesin status of the strain . P . aeruginosa isolates failed to agglutinate yeast and erythrocytes and also adhered poorly to buccal cells . A relationship between bacterial hydrophobicity and in vitro adherence was not found.

J Immunol, 1984 May, 132(5), 2590 - 2
An immunoprotective monoclonal antibody to lipopolysaccharide; Kirkland TN et al.; The ability of antisera against lipopolysaccharide (LPS) raised by immunization with gram-negative bacteria to prevent LPS toxicity and death from gram-negative bacteremia is well established . To demonstrate conclusively that the protective antibody is specific for LPS, we tested an anti-LPS monoclonal antibody (mAb) in three animal models . 7G is an IgG3 mAb directed against an oligosaccharide side chain determinant of LPS from E . coli 0111:B4 . This anti-LPS mAb increased the LD50 of 0111:B4 LPS in mice and protected rabbits against the dermal Shwartzman reaction elicited by 0111:B4 LPS . 7G mAb also protected mice against lethal infection with mucin-enhanced E . coli 0111:B4 . Pretreatment with 250 micrograms of 7G increased the LD50 by more than 1.5 logs . These studies prove that oligosaccharide side chain-specific antibody to LPS confers protection against LPS toxicity in vivo and against experimental gram-negative infection . In addition, these studies suggest the potential of anti-LPS monoclonal antibody as therapy for gram-negative infection.

J Biol Chem, 1984 Apr 25, 259(8), 4852 - 9
The biosynthesis of gram-negative endotoxin . Formation of lipid A disaccharides from monosaccharide precursors in extracts of Escherichia coli; Ray BL et al.; We have discovered an enzyme in the cytosol of Escherichia coli that generates lipid A disaccharides from monosaccharide precursors by the following route: 2,3-diacyl-GlcN-1-P + UDP-2,3-diacyl-GlcN---- 2,3-diacyl-GlcN (beta, 1----6) 2,3-diacyl-GlcN-1-P + UDP . Previous studies from our laboratory have documented the presence in vivo of the precursors 2,3-diacylglucosamine 1-phosphate (2,3-diacyl-GlcN-1-P) (lipid X of E . coli) and UDP-2,3-diacylglucosamine (UDP-2,3-diacyl-GlcN) (Bulawa, C.E., and Raetz, C.R.H.J . Biol . Chem . 259, 4846-4851) . Both substrates are novel glucosamine-derived phospholipids, acylated with beta-hydroxymyristoyl moieties, and they accumulate in E . coli mutants defective in the pgsB gene . Synthetic ADP-, GDP-, and CDP-2,3-diacylglucosamines are inefficient substrates compared to the naturally occurring UDP derivative . The free-acid form of the tetraacyldisaccharide 1-phosphate product (C68H129N2O20P) that is generated in vitro has Mr = 1325.74 as judged by fast atom bombardment mass spectrometry . Mild acid hydrolysis (0.1 M HCl for 30 min at 100 degrees C) liberates greater than 95% of the phosphate moiety as Pi . Detailed analysis by 1H and 13C NMR spectroscopy confirms the presence of a phosphate residue at position 1 of the disaccharide, an alpha-anomeric configuration at the reducing end, and a beta, 1----6 linkage between the two glucosamines . Importantly the disaccharide 1-phosphate synthase is missing in extracts of E . coli strains harboring the pgsB1 mutation, consistent with the massive accumulation of 2,3-diacyl-GlcN-1-P and UDP-2,3-diacyl-GlcN in vivo . The enzymatic reaction reported here represents a major biosynthetic route for the formation of lipid A disaccharides in E . coli and other Gram-negative bacteria . An in vitro system for the biosynthesis of lipid A disaccharides has not been described previously.

J Chromatogr, 1984 Apr 24, 288(2), 399 - 413
Capillary gas chromatography-mass spectrometry of carbohydrate components of legionellae and other bacteria; Walla MD et al.; Capillary gas chromatography using fused-silica columns followed by electron impact or chemical ionization mass spectrometry was used to profile and identify neutral and amino sugars present in several legionellae and other bacteria . A modified alditol acetate derivatization method was employed to produce volatile carbohydrate derivatives . Muramic acid, a component of bacterial peptidoglycan, was detected in all legionellae examined . Heptose, a component of bacterial lipopolysaccharide, was identified in Escherichia coli organisms and in several purified Gram-negative bacterial lipopolysaccharides but not in the legionellae examined . Two amino dideoxyhexoses were found to be present in several of the Legionellae examined . The potential of gas chromatography-mass spectrometry for the direct chemical characterization of microorganisms is discussed.

Lancet, 1984 Apr 7, 1(8380), 782 - 3
Clinical and microbiological features of Branhamella catarrhalis bronchopulmonary infections; Slevin NJ et al.; Branhamella catarrhalis bronchopulmonary infection was diagnosed in 101 patients with clinical lower respiratory tract infections by the presence of gram-negative intracellular diplococci in sputum and growth of more than 20 colonies of B catarrhalis in quantitative culture at a 10(-7) dilution . 94 patients had either chronic chest disease or were current or previous smokers, 59 had a cause of generalised immunosuppression, and 17 had a high risk of aspiration from the oropharynx . The pathogenicity of B catarrhalis was evident from purulence of sputum, fever, blood leucocytosis, and patchy pulmonary shadowing on chest radiographs . B catarrhalis infection contributed to 4 of 6 deaths . B catarrhalis was the only bacterial pathogen isolated from the sputum of 71 patients and it was isolated with other recognised bacterial pathogens from 30 patients . All 10 isolates of B catarrhalis tested were sensitive to oxytetracycline, all 82 tested were sensitive to cefuroxime, 93 of 96 were sensitive to erythromycin, and 85 of 95 to cotrimoxazole . Beta-lactamase was produced by 38 of 99 isolates of B catarrhalis.

Drug Intell Clin Pharm, 1984 Apr, 18(4), 314 - 6
Hypoprothrombinemia associated with cefoperazone treatment; Cristiano P; A patient with acute renal failure and gram-negative septicemia developed hypoprothrombinemia during treatment with cefoperazone . The coagulation defect was corrected by vitamin K administration . A multifactorial pathogenetic mechanism of vitamin K deficiency that developed during treatment with parenteral antibiotics is presented.

Arch Surg, 1984 Apr, 119(4), 379 - 83
Postoperative pneumonia . Determinants of mortality; Martin LF et al.; Postoperative pneumonia continues to be a major cause of mortality on surgical services . The determinants that affect survival in patients in whom postoperative pneumonia develops are not clearly defined . We completed a retrospective analysis of 136 patients in whom postoperative pneumonia developed after they had major operative procedures between 1974 and 1980 . These patients represented 1.3% of all operative cases, yet comprised 10% of the total 614 patients who died during the study period . The average age of the patients in whom pneumonia developed was 66 years . Significant determinants of death by chi 2 analysis included gram-negative pneumonitis, emergent operation, respirator-acquired pneumonia, postoperative peritonitis, and several factors that suggested that host defenses were overwhelmed (remote organ failure, positive blood cultures, or spread of infection to the second lung) . We concluded that postoperative pneumonia is a disease of elderly patients and that survival depends on the ability of the surgeon to help the patient localize and resist the challenge presented by virulent gram-negative organisms.

Am J Infect Control, 1984 Apr, 12(2), 76 - 82
Effects of handwashing agent, handwashing frequency, and clinical area on hand flora; Larson E; Six hundred twenty-two isolates from 554 samples taken from hands of 103 hospital personnel and 50 controls were obtained over a mean period of 35 days . Eighty-five were obtained immediately before and after handwashing (HW), the remainder after HW only . The relationship of HW agent used, clinical area and job, and frequency of HW to rates of bacterial colonization and numbers and types of organisms isolated, particularly gram-negative bacteria, were evaluated . The HW agent used ( nonantiseptic , hexachlorophene-based, chlorhexidine-based, or iodophor) was significantly correlated with the number of isolates obtained from each sample . Control subjects, all of whom used nonantiseptic soaps, had 1.42 mean isolates per sample; hospital personnel who used nonantiseptic soap had a mean of 1.00 isolates per sample . Other means were 1.25, hexachlorophene; 1.43, iodophor; 0.79, chlorhexidine; and 0.67 for those who used several different antiseptics, p less than 0.0001 . The agent was also correlated with the type of organisms isolated (p = 0.002), but not with the counts of colony-forming units (CFU) . Frequency of HW was significantly correlated with CFU counts before (p = 0.03) and after (p = 0.001) HW . In general, numbers decreased with increasing HW frequency, but at the higher HW frequencies there was a slight rise . There were significant differences in numbers of isolates per sample according to clinical area, with personnel working in obstetrics and nonpatient areas having the greatest number and those working on neonatal and medical-surgical units having the least (p less than 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)

Am J Med, 1984 Apr, 76(4), 664 - 71
Symposium on infectious complications of neoplastic disease (Part II) . Immunoprophylaxis and serotherapy of bacterial infections; Young LS; Immunologic approaches to the prevention and treatment of bacterial infections date back to the antecedent century . Recent interest and controversy has centered about the efficacy of gram-negative bacillary vaccines, antiserum against endotoxin, and pneumococcal vaccines . Immunization of cancer patients with Pseudomonas lipopolysaccharide vaccines has yielded inconsistent results . Factors limiting the further application of this approach are the poor immune responses in neutropenic patients and the marked pyrogenicity and pain associated with vaccine administration . Similarly, patients being treated for neoplasms of the hematopoietic system are not likely to show good antibody responses to pneumococcal antigens, even though they are not toxic . Pneumococcal immunization appears to be effective, at least as measured in terms of antibody titers, in those patients with lymphoma who have not undergone splenectomy and are not receiving chemotherapy at the time of immunization . The most reliable approach towards immunoprophylaxis may be the passive one, with antibody being produced in normal donors . The antibodies are short-lived, and this type of prophylaxis still needs to be evaluated in controlled trials . In a recently completed controlled therapeutic trial, the therapeutic application of an antiserum against core endotoxin antigens resulted in a significant reduction in deaths and increased recovery from shock complicating gram-negative sepsis . However, antiserum failed to protect cancer patients or neutropenic subjects.

South Med J, 1984 Apr, 77(4), 458 - 61
Colonization of central venous catheters; Prager RL et al.; We studied etiologic factors important in colonization of 179 central venous catheters (CVCs) in patients randomized into group 1 (who received daily topical applications of povidone-iodine) or group 2 (who received only dry dressing changes) . Colonization rates of CVC tips were similar between group 1 (18/84 or 21%) and group 2 (22/95 or 23%) . Peripheral blood cultures grew Candida in eight hyperalimented patients (evenly divided between groups 1 and 2), S epidermidis in four other patients (also evenly divided), and gram-negative bacteria in three patients . Colonization rates for CVCs in place for 0 to seven days was 15.6% (17/109) and 76.7% (23/30) if used from eight to 30 days . Inflammatory signs at CVC sites were often absent when CVCs became colonized or produced bacteremia . Unimportant determinants of CVC colonization included skin securement of CVCs, antibiotic infusions through CVC lines, and masking and gowning of physicians before CVC placement . Daily applications of povidone-iodine did not reduce colonization of CVCs as compared to dry dressing changes.

Am J Med, 1984 Apr, 76(4), 645 - 51
Symposium on infectious complications of neoplastic disease (Part II) . Chemoprophylaxis of bacterial infections in granulocytopenic patients; Henry SA; Several prospectively randomized trials have shown that the administration of prophylactic oral nonabsorbable antibiotics may be beneficial in decreasing the incidence of infection in granulocytopenic patients, whereas others have not . Intolerable nausea and vomiting have prevented the prolonged use of these agents in some studies . Discontinuation of therapy while patients are still granulocytopenic has carried the risk of life-threatening infections, often with aminoglycoside-resistant gram-negative organisms . The benefit of selective decontamination with trimethoprim/sulfamethoxazole used prophylactically remains controversial . The use of trimethoprim/sulfamethoxazole may also be associated with the development of resistant, potentially pathogenic, organisms or prolonged neutropenia . These regimens do not appear to be indicated when patients are anticipated to be neutropenic for less than three weeks . Even in patients with prolonged neutropenia, the risks of such treatment must be weighed against potential benefits.

J Pediatr, 1984 Apr, 104(4), 495 - 500
Spontaneous bacterial peritonitis; Clark JH et al.; Spontaneous bacterial peritonitis should be considered in the evaluation of any patient with acute abdominal complaints, especially in the presence of preexistent ascites . Paracentesis is indicated in all suspected cases of spontaneous peritonitis in order to obtain necessary studies, including microbial cultures . Broad-spectrum antibiotic coverage has become necessary because of the increasing incidence of gram-negative isolates . Ascitic pH and lactate may provide accurate information in the evaluation of spontaneous peritonitis, although increased clinical awareness remains the key to proper diagnosis.

Arch Biochem Biophys, 1984 Apr, 230(1), 250 - 63
Purification and properties of gentamicin nucleotidyltransferase from Escherichia coli: nucleotide specificity, pH optimum, and the separation of two electrophoretic variants; Van Pelt JE et al.; Gentamicin nucleotidyltransferase, AAD 2", catalyzes the transfer of a nucleotide to many aminoglycoside antibiotics, which are the drugs of choice in the treatment of gram-negative bacterial infections . The transfer is accompanied by the production of pyrophosphate, which is coupled to three other enzymes so that an increase in absorbance at 340 nm of NADPH can be monitored continuously as a quantitative assay of activity . A purification method was developed for this enzyme using all common principles of protein purification . These include selection of a desirable source of enzyme (choice of plasmid pMY 10), maximizing cellular yield of enzyme (controlled and monitored growth of Escherichia coli pMY 10/W677), selective extraction of protein (modified osmotic shock), removal of nucleic acids (precipitation with streptomycin sulfate), concentration of protein (precipitation with ammonium sulfate), removal of low-molecular-weight impurities (chromatography on Bio-Gel P-2), separation of proteins on the basis of charge (ion-exchange chromatography on DEAE-Bio-Gel A), separation of proteins according to a biospecific property (affinity chromatography on gentamicin-Affi-Gel), and separation of proteins according to size (gel filtration on Ultrogel AcA 54) . Purification to near-homogeneity revealed the presence of two related forms of enzyme . The first had a specific activity of 0.134 units/mg, bound rapidly and tightly to gentamicin-Affi-Gel, eluted as a function of ionic strength from Ultrogel, and migrated faster during electrophoresis in both the presence and absence of sodium dodecyl sulfate . It has an isoelectric point of 5.7 +/- 0.2 and consists of a single polypeptide of 32,500 Da . Kinetic characterization showed a pH optimum of 9.5 and Michaelis constants of 2.76 +/- 0.35 microM for tobramycin, 404 +/- 28 microM for Mg-ATP, 2008 +/- 260 microM for Mg-CTP, 30 +/- 3 microM for Mg-dATP and Mg-dGTP, and 90 +/- 7 microM for Mg-dCTP and Mg-dTTP . The second form had a specific activity of 0.274 unit/mg . It also bound tightly to gentamicin-Affi-gel but the onset of binding was time dependent . This form migrated slower during polyacrylamide gel electrophoresis in both the presence and absence of sodium dodecyl sulfate . It has an isoelectric point of 6.0 +/- 0.2 and consists of a single polypeptide of 31,500 Da . The exact relationship between the two forms has not been elucidated . It is probable that they have a recent common ancestor or are the same polypeptide because the amino acid compositions and polypeptide chain lengths are essentially identical.(ABSTRACT TRUNCATED AT 400 WORDS)

Drug Intell Clin Pharm, 1984 Apr, 18(4), 308 - 9
Isotretinoin: a reappraisal; Conner CS; Isotretinoin is remarkably effective in the treatment of severe cystic acne, however, many complications have been observed during treatment and new toxic effects have been reported . Hypertriglyceridemia associated with decreases in high density lipoprotein (HDL) cholesterol has occurred in 25 percent of patients and requires monitoring during treatment . Painful erosions with granulation tissue recently have been reported in patients with severe acne . Other complications have included corneal opacities, pseudotumor cerebri, hypercalcemia, photosensitivity reactions, abnormal liver function tests, and skeletal hyperostosis . Isotretinoin is teratogenic and should be avoided during pregnancy . With the increasing acceptance and use of isotretinoin for cystic acne, as well as related disorders (inflammatory papulopustular acne with scarring, gram-negative folliculitis, and acne rosacea), a reevaluation of isotretinoin aimed at reducing complications is in order . Patient selection criteria and guidelines directed at reducing these complications are presented.

Am J Med, 1984 Mar 30, 76(3A), 146 - 54
Opportunistic infections in severely burned patients; Pruitt BA Jr et al.; The risk of infection in burn patients, which is proportional to the extent of burn, reflects the combined effect of impairment of all aspects of the host defense system and microbial factors . The microbial flora colonizing the burn wound changes with time following injury and provides the organisms causing infections in burn patients . The temporal pattern of the predominant gram-negative organisms causing infections in a burn unit resembles that of a succession of mini-epidemics necessitating an active program of microbial surveillance to guide treatment of infections . Topical chemotherapy has significantly reduced the occurrence of invasive burn wound infections, but microbial control is imperfect and the burn wound, as well as the patient as a whole, must be closely monitored (using wound biopsies as indicated) to diagnose and treat infection in a timely manner . The treatment of burn wound infections is guided by extent and depth of microbial invasion, density of microorganisms, and systemic changes . As a manifestation of immunologic impairment, infection in sites other than the burn wound remains the most frequent cause of death in burn patients . The use of broad spectrum serologic agents to enhance immuno-competence in extensively burned patients may reduce the occurrence of life threatening opportunistic infections.

Med J Aust, 1984 Mar 3, 140(5), 286 - 7
How zealously should a patient with relapsed acute myeloid leukaemia be treated? Good survival after five simultaneous, potentially lethal, complications; Lowenthal RM; A relapse of acute myeloid leukaemia occurred in a 45-year-old woman 18 months after the disease was initially diagnosed and treated . During remission reinduction therapy, she developed a Gram-negative septicaemia, acute respiratory failure, acute renal failure, diabetic hyperglycaemia with ketoacidosis, and probable bacterial meningitis . She required assisted respiration for two days, received peritoneal dialysis for five days, and was unconscious for seven days . The patient eventually recovered, achieved full remission of her leukaemia, and survived a further 2 1/2 years, mostly in excellent health . Oncologists are often criticized for unjustifiable optimism and excessive zeal in the treatment of patients with malignant disease . This case illustrates that such optimism and zeal may be justified, and that intensive efforts to save the lives of seriously ill patients with chemosensitive malignant diseases are worthwhile.

Cutis, 1984 Mar, 33(3), 281 - 2
Recalcitrant unilateral infection associated with congenital leg hypertrophy cleared by control of hyperhidrosis; Shelley WB et al.; A man with congenital hypertrophy of the left leg experienced concomitant fungal and gram-negative bacterial infection of the foot on the same side . Oral antibiotic therapy failed, compromised by a post-surgical short bowel . Despite two hospitalizations for intensive care with intravenous antibiotic therapy, the infection did not clear until his plantar hyperhidrosis on the affected side was eliminated by the use of topical aluminum chloride.

Biomed Mass Spectrom, 1984 Mar, 11(3), 132 - 41
Laser desorption mass spectrometry of synthetic lipid A-like compounds; Seydel U et al.; The applicability and the present limitations of the laser microprobe mass analyser LAMMA -500 as an instrument for the structural analysis of higher molecular weight, non-volatile, bio-organic compounds (less than or equal to 2000 u) were investigated . For this purpose mass spectra of various synthetic and natural compounds representing cell wall components of Gram-negative bacteria, e.g . phospholipids and lipid A-like molecules were studied . In several cases these spectra exhibited relatively simple and interpretable patterns with a prominent quasi-molecular ion originating from alkali attachment . For one group of the compounds studied--synthetic lipid A-like molecules containing a phosphate moiety--the spectra were rather complicated and lacked pronounced quasi-molecular peaks . Possible reasons for this observation are discussed.

J Infect Dis, 1984 Mar, 149(3), 443 - 8
The association of aminoglycoside plasma levels with mortality in patients with gram-negative bacteremia; Moore RD et al.; To determine the association of aminoglycoside levels with mortality from gram-negative bacteremia, we analyzed the case reports of patients from four prospective, randomized, and controlled clinical trials of gentamicin, tobramycin, and amikacin . Twelve (13.5%) of 89 patients died . One (2.4%) death occurred in 41 patients with early (1-hr postinfusion) peak concentrations of greater than 5 micrograms/ml of gentamicin and tobramycin and of greater than 20 micrograms of amikacin/ml; nine deaths (20.9%) occurred in 43 patients with lower concentrations . Five (8.3%) deaths occurred in 60 patients with mean peak concentrations for the entire course of therapy of greater than 5 micrograms/ml of gentamicin and tobramycin and of greater than 20 micrograms of amikacin/ml; five (20.8%) deaths occurred in 24 patients with lower concentrations . Stepwise discriminant analysis showed that therapeutic early peak concentration was a significant factor in the presence of three other factors: severity of underlying illness, peak temperature, and initial leukocyte count . The results suggest the importance of achieving adequate early aminoglycoside levels in patients with gram-negative bacteremia.

Am J Med, 1984 Mar, 76(3), 429 - 35
Significance of serum bactericidal activity in gram-negative bacillary bacteremia in patients with and without granulocytopenia; Sculier JP et al.; Serum bactericidal activity was determined routinely in 89 patients with gram-negative bacillary bacteremia, 79 of whom were analyzed because they had granulocyte counts either below 100/mm3 or above 1,000/mm3 . A peak (one hour after the administration of the antibiotics) serum bactericidal titer of 1:8 or more in non-granulocytopenic patients or 1:16 or more in severely granulocytopenic patients could be correlated with a favorable clinical response, in 98 percent (44 of 45) (p less than 0.0001) and 87 percent (20 of 23), (p less than 0.001) respectively . Granulocytopenic patients required a statistically significantly higher serum bactericidal activity for a favorable response . Serum bactericidal activity appears to be a useful and simple method to monitor antibiotic treatment in gram-negative bacillary bacteremia, especially when combination therapy is used.

Am J Dis Child, 1984 Mar, 138(3), 240 - 2
Vitamin K-dependent coagulation factors in gram-negative septicemia; Corrigan JJ Jr; Coagulation studies were performed on 16 children with gram-negative septicemia without the complications of septic shock, liver disease, malnutrition, or laboratory evidence of classic disseminated intravascular coagulation (DIC) . Ten (63%) of the 16 cases were found to have abnormal partial thromboplastin and/or prothrombin times . The coagulopathy was caused by a reduction in the vitamin K-dependent coagulation factors . The mechanism that produced this coagulopathy was not known, but evidence was found that suggested that endotoxin may interfere with the vitamin K-carboxylation reaction . The data indicated that abnormal coagulation screening test results in children with gram-negative septicemia were not specific for DIC and that a significant number of patients had a coagulopathy not related to DIC.

Ann Intern Med, 1984 Mar, 100(3), 333 - 8
A new Legionella species, Legionella feeleii species nova, causes Pontiac fever in an automobile plant; Herwaldt LA et al.; From 15 to 21 August 1981, Pontiac fever affected 317 automobile assembly plant workers . Results of serologic tests were negative for Mycoplasma, Chlamydia, respiratory tract viruses, and previously described legionellae . A gram-negative, rod-shaped organism (WO-44C) that did not grow on blood agar, required L-cysteine for growth, and contained large amounts of branched-chain fatty acids was isolated from a water-based coolant . The organism did not react with antisera against other legionellae, and on DNA hybridization the organism was less than 10% related to other Legionella species . Geometric mean titers found by indirect fluorescent antibody testing to WO-44C were significantly higher in ill employees than in controls (p = 0.0001) . Attack rates by department decreased linearly with the department's distance from the implicated coolant system . The etiologic agent apparently was a new Legionella species; we propose the name Legionella feeleii species nova (AATC 35072) . This is the first outbreak of nonpneumonic legionellosis in which the etiologic agent is not L . pneumophila, serogroup 1.

Boll Ist Sieroter Milan, 1984 Mar, 63(1), 57 - 60
Transient symptomatic cryoglobulinemia in gram-negative bacteria infections; Galli M et al.; Mixed polyclonal cryoglobulinemia was observed in seven patients suffering from severe gram-negative bacterial infections and/or septicaemias and presenting with arthralgia and purpuric manifestations on admission . Cryoglobulins disappeared after recovery from infection in all of them and were never found during a longterm follow-up . In our opinion, gram-negative bacteria may induce the synthesis of cryoglobulins via a non-specific T-independent B-cell stimulation triggered by cell-wall lipopolysaccharides . This kind of infections, especially if chronic or relapsing, might play a role in the aetiology of some so-called essential mixed cryoglobulinemias.

Am J Physiol, 1984 Mar, 246(3 Pt 2), H344 - 50
Cardiopulmonary response of the rat to gram-negative bacteremia; Pass LJ et al.; Hemodynamic and respiratory effects of a continuous 5-h intravenous infusion of live Escherichia coli were studied in rats . Control animals were infused with saline . Rats infused with 1.8 +/- 0.4 X 10(10) bacteria/h did not survive a 5-h infusion . These animals developed early hypotension and reduced cardiac output (CO) measured by thermal dilution technique . Rats infused with 8.0 +/- 0.4 X 10(9) bacteria/h survived a 5-h infusion with hypotension and reduced CO occurring later in the course of bacteremia . Heart rate was markedly elevated in both septic groups . Arterial blood gas measurements revealed that partial pressure of O2 was not affected by bacteremia, but partial pressure of CO2 was significantly decreased . Arterial pH remained within the normal range indicating respiratory compensation of a metabolic acidosis . Since hypotension and reduced CO were accompanied by a fall in right atrial pressure (RAP) during bacteremia, a third septic group was studied to evaluate cardiac performance during volume loading . After 3-5 h of bacteremia, a 40% reduction in CO was associated with a significant drop in arterial pressure and RAP . Despite volume loading, ventricular stroke work and arterial pressure were significantly reduced compared with control animals . The results indicate that severe gram-negative bacteremia produces myocardial depression in the rat . This model can be useful for further studies of cardiac dysfunction during sepsis.

J Hosp Infect, 1984 Mar, 5(1), 29 - 37
Pseudomonas septicaemia . A review of 60 cases observed in a university hospital; Lechi A et al.; Epidemiological, clinical features and the pathogenesis of 60 cases of pseudomonas septicaemia, observed over a period of 7 years from 1975 to 1981, are described . The mean frequency of occurrence was 0.40 episodes per 1000 admissions and an incidence of 66 per cent was observed in patients with serious underlying diseases, such as haematological malignancies and neoplasia . Thirty-seven patients had received cytotoxic and immunosuppressive therapy and showed a marked leukopenia, and whenever the leukocyte count fell below 2000/mm3, the prognosis was significantly worse . The most common portals of entry were the respiratory and gastrointestinal tracts . The overall mortality was very high (75 per cent) and mainly related to septic shock . Apart from the very high frequency of this complication (24 patients, 40 per cent of all cases), a typical clinical picture, distinguishing pseudomonas from other Gram-negative septicaemias, did not emerge.

J Infect Dis, 1984 Mar, 149(3), 320 - 9
Promiscuous transfer of drug resistance in gram-negative bacteria; Guiney DG Jr; Bacterial conjugation is a major mechanism for the spread of antibiotic-resistance genes in pathogenic organisms . In gram-negative bacteria, broad-host-range drug-resistance plasmids mediate genetic exchange between many unrelated species . The mechanism of conjugation encoded by the broad-host-range IncP plasmid RK2 has been studied in detail . The location and sequence of the transfer origin of RK2 has been determined . Several barriers limit plasmid transfer between unrelated bacteria: interactions at the cell surface may prevent effective mating contact, restriction systems may degrade foreign DNA, or the plasmid may not replicate in the new host . RK2 has evolved specific mechanisms by which it overcomes these barriers; this plasmid can mediate the transfer of resistance to most gram-negative bacteria.

J Dairy Res, 1984 Feb, 51(1), 59 - 67
Origins and levels of post pasteurization contamination of milk in the dairy and their effects on keeping quality; Schroder MJ; Bacterial post pasteurization contamination with psychrotrophic Gram-negative rods (GNR) was measured in commercial milks before and after transfer to retail containers . The tanks of pasteurized milk feeding the filling units contaminated milk less often (39% of samples) but usually at a higher level than the filling units (92% of samples) . The number of GNR present had a considerable influence on the shelf life of milk, and the range found in commercially pasteurized milk was reflected in a wide range of shelf lives.

J Clin Microbiol, 1984 Feb, 19(2), 116 - 21
Comparison of Legionella pneumophila, L . micdadei, L . bozemanii, and L . dumoffii by transmission electron microscopy; Hebert GA et al.; Clinical isolates of Legionella pneumophila, L . micdadei, L . bozemanii, and L . dumoffii were grown on charcoal-yeast extract agar from a living-medium inoculum and prepared for transmission electron microscopy by three different methods . Cells of all four Legionella species possessed cytoplasmic vacuoles, a gram-negative type of cell envelope with a dense peptidoglycan-like layer, a ruthenium red-positive polysaccharide capsule, and a single subpolar flagellum . The dense polysaccharide capsule seen on cells of L . micdadei was separated from the outer membrane by an extra layer of electron-lucent material that was not present on cells of the other species examined.

Postgrad Med, 1984 Feb 1, 75(2), 177 - 86
Aminoglycoside therapy . Improving patient response and safety; Ackerman BH et al.; The aminoglycosides have long been effective antibiotics in treatment of serious gram-negative infections . However, ototoxicity and nephrotoxicity have been of major concern because of the narrow therapeutic range of these agents and the wide variability in pharmacokinetics among patients . With recent advances in aminoglycoside monitoring techniques, the risk of toxicity has been greatly reduced . Automated methods are now available to quickly and precisely measure aminoglycoside concentration in serum, and by applying pharmacokinetic principles to serum concentration-time data, the clinician can readily calculate the correct dosage for each patient . Programs for computers and programmable calculators, also now available, can assist in these calculations.

J Lab Clin Med, 1984 Feb, 103(2), 284 - 93
Plasma lactoferrin reflects granulocyte activation via complement in burn patients; Wolach B et al.; Complement activation and neutropenia have been observed in thermally injured animals . In burn patients, granulocyte chemotaxis and morphological loss of specific granules occur . We conjectured that complement is activated in humans and, in turn, induces granulocytes to secrete lactoferrin (LF), a marker of granulocyte activation . Twenty burn patients were evaluated for absolute granulocyte count (AGC), plasma levels of anaphylatoxins (C3a, C4a, C5a), and lactoferrin . The AGC directly correlated with the extent of the burn on day 1 . Similarly, plasma LF on day 1 correlated with the percent burn . Those with greater than 30% burn had plasma LF between 10 and 40 micrograms/ml (normal LF = 1.5 +/- 1.8 micrograms/ml) . In five patients without further complications followed serially, plasma LF did not return to normal until 2 to 5 weeks . In all patients, there was evidence of complement activation; C4a ranged between 283 and 13,064 ng/ml and C3a between 19 and 852 ng/ml . In some patients, C5a was detectable, but the values correlated inversely with the extent of burn . On the other hand C3a and C4a levels did not correlate with the extent of burn but threefold to fivefold rises of C3a levels on days 7 and 9 predicted gram-negative sepsis . Although plasma LF did not predict sepsis, levels greater than 12 micrograms/ml on day 1 heralded the onset of neutropenia on day 3 in 60% of patients with 30% burn . Six of 20 patients developed pulmonary radiographic changes and, in five of the six, the changes occurred by day 3 . Plasma LF in all six patients on day 1 was greater than 17 micrograms/ml . In two patients with greater than 50% burn, depletion of granulocyte LF was demonstrated histochemically . These studies indicate that complement is activated in burn patients, which is associated with granulocyte secretion . Measurement of plasma anaphylatoxins and LF may serve as useful aides in clinical management of these patients.

Burns Incl Therm Inj, 1984 Feb, 10(3), 217 - 24
Correlation of quantitative burn wound biopsy culture and surface swab culture to burn wound sepsis; Tahlan RN et al.; The present study includes seventeen patients with second and third degree fresh burns involving 15-50 per cent total body surface area (TBSA) . Surface swabs and quantitative burn wound biopsy cultures were obtained during postburn weeks 1, 2 and 3 and correlation was studied . To obtain bacterial counts the technique described by Loebel et al . (1974) was used . The patients were divided in two groups depending upon burn body surface area involved . The first group includes five patients with burns between 15-29 per cent body surface area and the second group includes the rest of the twelve patients with burns between 30-50 per cent body surface area . No patient from group I showed any sign or symptom of sepsis whereas seven patients from group II developed sepsis and three died . These three patients showed positive blood culture at the time of death . Of the 48 cultures obtained in all the patients over 3 weeks, 7 cultures showed differences between swab and biopsy cultures . Genticyn was the most effective drug against Gram-negative organisms.

Dig Dis Sci, 1984 Feb, 29(2), 171 - 7
Perforated diverticulum of the terminal ileum . A previously unreported cause of suppurative pylephlebitis and multiple hepatic abscesses; Navarro C et al.; We describe the case of a 58-year-old man who presented to the hospital with central abdominal pain, nausea, fever, chills, and dyspnea . While in the hospital, jaundice appeared and the liver function tests revealed features of both cholestasis and hepatocellular injury . He developed gram-negative septicemia and died on the sixth hospital day . Autopsy disclosed a perforated terminal ileal diverticulum and a contiguous mesenteric abscess . There was also severe phlebitis of mesenteric venous radicles which extended superiorly to the intrahepatic portal venules and veins . The portal veins were surrounded by multiple hepatic abscesses that varied in size from microscopic to 2.5 cm . This appears to be the first report in the world literature of suppurative pylephlebitis and hepatic abscesses resulting from a perforated ileal diverticulum . The subject of small bowel non-Meckelian diverticulosis is reviewed because of the rarity of this condition and the diagnostic challenges it poses.

Infect Immun, 1984 Feb, 43(2), 684 - 92
Degree of antibody-independent activation of the classical complement pathway by K1 Escherichia coli differs with O antigen type and correlates with virulence of meningitis in newborns; Pluschke G et al.; A total of 95 K1 Escherichia coli strains of the O (lipopolysaccharide) serotypes O1, O7, or O18 had been analyzed previously for the ability to cause bacteremia after colonizing the gut of newborn rats . In this study, these strains were tested for their resistance to the bactericidal activity of rat serum . All strains that had caused bacteremia in a high percentage of the inoculated rats were able to survive for several hours in 90% adult rat serum . With only a few exceptions, O7:K1 and O18:K1 strains were serum resistant and virulent, whereas O1:K1 strains were serum sensitive and avirulent . Serum sensitivity was due to the classical complement pathway . K1 strains of all three O serotypes were resistant to the alternative complement pathway . O7:K1 and O18:K1 cells were killed efficiently after the classical pathway was triggered by specific antilipopolysaccharide antibodies . However, killing of O1:K1 bacteria by the classical pathway system did not require antibodies . Isolated O1-lipopolysaccharide fixed complement more efficiently than did isolated O7- or O18-lipopolysaccharide, suggesting that the differences in the chemical structure of the O antigens are responsible for the observed differences in complement sensitivity . In combination with epidemiological data, the results indicate that antibody-independent classical pathway activation provides an important defense mechanism for newborns against certain gram-negative infections.

Cancer, 1984 Jan 15, 53(2), 219 - 23
Indwelling venous access catheters in patients with acute leukemia; Reilly JJ Jr et al.; Reliable venous access is often a serious problem in the cancer chemotherapy patient . In one year, the authors of this report have inserted 26 chronic double-lumen silastic Hickman catheters in 25 acute leukemia patients . Each patient received an average 12 courses of combination chemotherapy, 11.5 packed red blood cell units, 48.0 platelet units, 4.2 fresh frozen plasma units, and numerous antibiotic doses via the catheters, which remained in situ 101 +/- 97.4 days . Complications included early hemorrhage in two cases, and catheter occlusion in six . Four catheters were removed for occlusion . Fourteen patients suffered bacteremia, predominantly due to gram-negative organisms; six catheters were removed for persistent sepsis . The remaining catheters remained functional until death or elective removal . Eight patients are currently alive as outpatients with functional catheters . The Hickman catheter effectively achieves reliable venous access in the cancer patient . The risks of catheter sepsis must be considered carefully in the immunosuppressed, leukopenic patient.

Minerva Med, 1984 Jan 14, 75(1-2), 1 - 4
{Infectious endocarditis . Current aspects of its epidemiology and occurrence}; Lenti G et al.; In epidemiological terms the frequency of infectious endocarditis is constant . In contrast acute forms in males and old people are on the increase . Atypical cases are more frequent than in the past so that prompt diagnosis is often difficult . Aetiologically, mycete and gram-negative cases are increasingly common.

Clin Orthop, 1984 Jan-Feb, (182), 117 - 26
Factors influencing the incidence and outcome of infection following total joint arthroplasty; Poss R et al.; During a ten-year period, 4240 total hip, knee, and elbow arthroplasties were performed . The overall infection rate was 1.25% . Certain groups were identified as being at higher risk of infection following total joint arthroplasty: rheumatoid arthritics were at 2.6 times greater risk than osteoarthritics; patients undergoing total hip arthroplasty as a revision of a previous operation were eight times more likely to have infection than those undergoing a primary operation; and patients with metal-to-metal hinged knee prostheses, when compared with patients with metal-to-plastic knee prostheses, were 20 times more likely to have infection . The majority of infections could be attributed either to perioperative problems or late bacterial seeding from a distant site . Although most infections occurred by two years after operation, late infections, particularly in rheumatoid patients via the hematogenous route, occurred as long as nine years after operation . There was no correlation between the Gram's-staining characteristics of the pathogen and the outcome of the infected joint . Gram-negative organisms were frequent in the perioperative period and reflected either nosocomial infection or the ineffectiveness of the prophylactic antibiotic regimen used in inhibiting gram-negative pathogens . The major factors that influenced the outcome of the infected joint included the interval from the initial surgery to recognition of infection, the delay in institution of appropriate treatment, the particular joint that was infected, the integrity of the bone-cement interface, the type of prosthesis used, and the host susceptibility . Identification of high-risk groups and the recognition that patients with joint implants are at risk of infection at any time in the postoperative period may lead to a lowered infection rate in the future.

Br J Dis Chest, 1984 Jan, 78(1), 66 - 74
Immunological abnormalities in patients with chronic bronchial suppuration: a possible relationship with endotoxaemia; Horan MA et al.; Chronic bronchial suppuration has been associated with elevations in immunoglobulin levels, circulating autoantibodies and immune complexes but the cause is obscure . In order to investigate one possible explanation we studied eight male subjects (mean age 54.9 years, range 39-68) and 16 female subjects (mean age 45.2 years, range 24-68) with bronchial suppuration attributed to bronchiectasis . Gram-negative organisms were found in the sputum of 95% of subjects . Forty per cent (10 subjects) had elevations in immunoglobulin levels and 60% (15 subjects) had circulating autoantibodies . Eighty-eight per cent (21 subjects) had mild-moderate increases in the level of C-reactive protein . Endotoxin was detected using the limulus amoebocyte lysate test in 50% (12 subjects) . Endotoxin was found only in patients with an immunological abnormality and was more likely to be detected in those with both elevated immunoglobulins and circulatory autoantibodies . It is suggested that the endotoxin may be of aetiological significance in the development of autoantibodies and elevations of immunoglobulin levels.

Am Surg, 1984 Jan, 50(1), 43 - 8
Synthetic vascular graft infection . The continuing headache; Bunt TJ et al.; Twenty synthetic vascular graft infections are reviewed . Diagnosis of aortic shaft infections has been improved by combined CAT and Gallium scanning, with a 100 per cent sensitivity and specificity . An increasing incidence (70%) of gram-negative or resistant infections is noted, with a correspondingly high mortality . Delay in diagnosis resulted in an 100 per cent mortality versus 20 per cent when diagnosis was made rapidly . Mortality was usually from sepsis, but nearly half of the deaths occurred due to stroke or myocardial infarction.

Ann Otol Rhinol Laryngol, 1984 Jan-Feb, 93(1 Pt 1), 52 - 6
Experimental otitis media with effusion following middle ear inoculation of nonviable H influenzae; DeMaria TF et al.; In order to test the hypothesis that nonviable bacteria can induce middle ear inflammation leading to persistent middle ear effusion (MEE), we conducted an animal experiment using formalin-killed Hemophilus influenzae, the bacterium reported to be the most common pathogen isolated from chronic MEEs . Over 70% of the chinchillas injected with formalin-killed H influenzae type b or a nontypeable isolate developed sterile, straw-colored serous MEEs, and exhibited histological evidence of extensive inflammatory changes of the middle ear mucosal connective tissue and epithelium . Control animals injected with pyrogen-free sterile saline did not exhibit any inflammatory changes or effusions in the middle ears . Our data suggest that endotoxin on the surface of H influenzae, a gram-negative bacterium, may be responsible for the induction of the otitis media with effusion . It is suggested that endotoxin (even when the organisms are no longer viable) may be responsible for the production of serous MEE and inflammatory changes in the middle ear.

Nephron, 1984, 36(1), 65 - 7
Treatment of peritonitis in continuous ambulatory peritoneal dialysis patients with co-trimoxazole; Glasson P et al.; Peritonitis in patients undergoing continuous ambulatory peritoneal dialysis (CAPD) represents the most frequent and difficult problem related to this new form of treatment of ESRD patients . Various treatments have been reported previously . The aim of this study was to investigate the efficiency of a standardized initial treatment in 45 episodes of peritonitis . This was designed to be rapidly efficient, devoided of side-effects and easy enough to be performed by the patients themselves . When peritonitis was clinically suspected, patients received intraperitoneal co-trimoxazole (80 mg trimethoprim, 400 mg sulfamethoxazole), in each of the four daily bags concomitantly with 1,000 U heparin during 2 weeks and half of this dose during 2 other weeks . Our results demonstrate that 88% of the isolates were sensitive to co-trimoxazole and 85% of the patients completed this treatment . All were cured and no relapses were observed . Only 18 days of hospitalisation were required in the 45 episodes of peritonitis . Another anti-infective agent was used in 3 cases of gram-negative peritonitis and 4 other initially resistant to co-trimoxazole . It is concluded that initial treatment of CAPD peritonitis with co-trimoxazole is justified by the high proportion of sensitive germs and that it represents a safe, efficient and inexpensive treatment.

Scand J Urol Nephrol Suppl, 1984, 86, 71 - 4
Gas chromatographic analysis of cellular fatty acids in anaerobic curved bacteria isolated from the vagina; Skarin A et al.; The fatty acid compositions of forty-five strains of anaerobic curved rods isolated from vaginal content were analysed by gas chromatography . Twenty-six of the strains were of a short Gram-variable type and 19 of a long Gram-negative type . The bacteria were subjected to alkaline saponification, after which the liberated fatty acids were methylated using boron trichloride-methanol . The samples were introduced onto a cross-linked (SE-54) fused silica capillary column . The major peaks in the chromatograms from all strains corresponded to C14:0, C16:0, C18:2, C18:1 and C18:0 fatty acids . Five additional components were seen, three of them identified by mass spectrometry as fatty aldehydes (saturated and unsaturated) of 16 to 18 carbon atoms, whereas the remaining two were not identified . The fatty aldehydes indicated the presence of plasmalogens in the bacterial strains studied . With two exceptions, all of the short rods were distinguishable from the long ones by the presence of one aldehyde and two unidentified compounds, all three eluting between C16:0 and C18:2.

Ophthalmology, 1984 Jan, 91(1), 60 - 3
Gram-negative corneal ulcers in elderly aphakic eyes with extended-wear lenses; Lemp MA et al.; We report 13 cases of corneal ulcers caused by gram-negative bacteria . These cases occurred in aphakic eyes wearing hydrophilic contact lenses on an extended-wear basis . The risks for infection in the elderly aphakic eye with extended-wear hydrophilic lenses are increased by delay in removal of the lens . Caution in the use of these lenses is urged . The ability of either the patient or an attendant to remove the lens as soon as redness, diminished vision, or pain presents itself is essential in minimizing the risks for serious infection.

Arzneimittelforschung, 1984, 34(7), 830 - 1
{Metronidazole concentration of the cerebrospinal fluid from slightly inflamed meninges}; Hoffmann HG et al.; The concentrations of metronidazole (Clont i.v.) in the cerebrospinal fluid were measured in 12 patients with viral meningitis or subsiding bacterial meningitis after a single infusion of 500 mg lasting 20 min . 1 h after infusion the CSF-concentrations were between 2,3 micrograms/ml and 7,4 micrograms/ml and 2 h after infusion between 6,5 micrograms/ml and 8,6 micrograms/ml . They attained 45,9% respectively 75,9% of the corresponding serum concentrations . Because the minimal inhibitory concentrations of the most important obligate anaerobic gram-negative bacteria are attained, it appears that metronidazole can be used for the treatment of bacterial meningitis caused by these pathogens.

Z Erkr Atmungsorgane, 1984, 163(3), 208 - 16
Gram-negative bacterial endotoxins in airborne dusts from washed and unwashed cottons; Olenchock SA; Gram-negative bacteria possess lipopolysaccharide-protein complexes termed endotoxins as an integral portion of their cell walls . These biologically active substances elicit profound effects both in vitro and in vivo . Organic dusts from many occupational environments contain measureable amounts of gram-negative bacteria and their endotoxins . In particular, cottons and cotton dusts contain these materials, and research is directed to elucidate the association between endotoxins and the acute and chronic respiratory effects of inhaling cotton dusts . It is the purpose of this paper to describe an experimental process which removes endotoxins from cotton by washing prior to processing the baled cottons . The studies show that water washing is a reliable method for removing endotoxins, and pulmonary function studies correlate well with endotoxin levels in the air . Quantification of airborne endotoxins provides a good indicator of cleanliness and toxocity of the cotton dusts . However, the differential toxicities of different endotoxins must be considered and evaluated further.

Biomed Pharmacother, 1984, 38(8), 397 - 403
{Bacteriologic analysis of 150 episodes of septicemia in a hematology department according to the length of hospitalization}; Durning A et al.; One hundred and fifty episodes of septicaemia in patients in a haematology service were analysed as a function of the date of onset in relation to the date of hospitalisation . One hundred and three patients of the 122 patients were granulocytopenic and 97 patients (65%) had less than 500 polymorphonuclear neutrophils per microliter . The septicaemic episodes were classified according to three time periods: septicaemia starting before admission and during the first 24 hours in hospital, septicaemia occurring between day 1 and day 8 and septicaemia beginning on day 9 or later . For each period different factors have been studied, the number of septicaemic episodes, the frequency of multiple infection, the severity of the granulocytopenia, the antibiotics used compared to their sensitivity in vitro, the outcome of the septicaemia, the organisms isolated by blood culture and their sensitivity to different antibiotics . The patients with septicaemia arriving at the hospital with a fever, and those starting during the first week of hospitalisation are finally comparable, even though the level of mortality was distinctly different (33 and 17% respectively) . The septicaemia starting after day 8 are more frequent and more severe (40% mortality) . These late onset septicaemias are characterised by a higher frequency of Gram negative hospital infections, a high frequency of mixed infection with bacteria resistant to several beta-lactams and aminoglycosides . The observation of these differences between these septicaemias occurring in the first week and those starting after the eighth day should be taken into account in the choice of an empirical antibiotic regimen . An association of 2 or 3 antibiotics for these two situations respectively, involving beta-lactams and aminoglycosides, is proposed by the authors.

Am J Emerg Med, 1984 Jan, 2(1), 45 - 59
Skeletal muscle metabolism and insulin resistance during endotoxin shock in the dog; Raymond RM; The effect of locally infused endotoxin on gracilis muscle glucose uptake was determined in anesthetized mongrel dogs . The effects of infusion of small amounts of Escherichia coli endotoxin into the arteries of isolated, innervated, constant flow perfused gracilis muscles on glucose uptake and other metabolic variables were determined . Locally infused endotoxin consistently caused a significant and substantial increase in skeletal muscle glucose uptake with no alterations in muscle arteriovenous difference of insulin, oxygen, carbon dioxode, or pH, or in venous blood hematocrit or temperature . These data demonstrate that endotoxin can act locally to increase glucose uptake by skeletal muscle, independent of the action of insulin or other metabolic factors . During natural (free flow) conditions, glucose uptake by the muscle increased markedly during six hours of shock . Increased glucose uptake occurred concomitantly with muscle ischemia and hypoxia . However, when muscle blood flow was held constant, thereby preventing local muscle ischemia and hypoxia, glucose uptake by the gracilis muscle did not change during shock . These results implicate local muscle ischemia and/or hypoxia as the mediator(s) of the increased muscle glucose uptake during shock . Further studies demonstrated that local muscle hypoxia was the stimulus for increased glucose uptake by skeletal muscle during endotoxin shock, and muscle ischemia per se did not alter muscle glucose uptake . Since approximately 50% of body mass is composed of skeletal muscle, the contribution of this organ system to the hypoglycemia of endotoxin shock in the dog may be substantial . The ability of insulin to promote glucose diffusion into skeletal muscle before and during gram-negative endotoxin shock was studied in mongrel dogs anesthetized with sodium pentobarbital . The in vivo, isolated, innervated, constant flow perfused gracilis muscle preparation was used . Prior to shock induction, close intra-arterial insulin infusion resulted in a 320% increase in muscle glucose uptake . However, at one, two, and three hours of endotoxin shock, gracilis muscle glucose uptake was unaltered by insulin infusion . This loss of responsiveness to insulin occurred with no alteration in gracilis muscle oxygen uptake, muscle venous P02, or muscle blood flow . During control experiments, however, the muscle response to intra-arterial infusion of insulin (increased glucose uptake) was unaltered during the three-hour control period . These data demonstrate that skeletal muscle insulin resistance develops early and is maintained during three hours of endotoxin shock in the dog.

Curr Med Res Opin, 1984, 9(3), 213 - 8
Serum concentrations and penetration into prostate of mecillinam and ampicillin; Jeppesen N et al.; The absorption of the two antibiotic pro-drugs pivmecillinam and pivampicillin was investigated in 10 patients with normal or slightly impaired renal function . After a therapeutic dose of 200 mg pivmecillinam plus 250 mg pivampicillin, the mean peak concentration in serum was 2.3 micrograms mecillinam and 5.3 micrograms ampicillin per ml . The elimination half-life was approximately 2 hours for both antibiotics . Consequently, a ratio of 2:5 was maintained in serum during the excretion period and this ratio is within the range where optimal synergistic effect between the two antibiotics against many Gram-negative organisms is most likely to occur . Penetration of mecillinam and ampicillin into the prostate, determined in tissue homogenate obtained from 10 patients undergoing transurethral resection of the prostate showed that the antibiotics penetrated equally well with a serum tissue ratio of approximately 2:1.

Comp Immunol Microbiol Infect Dis, 1984, 7(2), 101 - 8
Use of isoelectric focusing of bacterial proteins for characterisation of strains of motile anaerobic "corroding" bacteria isolated from soft tissue infections in cats and dogs; Love DN et al.; Isoelectric focusing of bacterial proteins in 1 mm thick agarose gels and ultrastructural analysis of cell walls were used to facilitate characterisation of motile "corroding" bacteria from subcutaneous abscesses and pyothorax infections from cats and dogs . Some of the motile "corroding" rods had ultrastructural cell wall features identical with Wolinella recta described from human periodontal pockets . Other strains had the more conventional cell wall ultrastructure of Gram negative bacteria . Isoelectric focusing of bacterial proteins divided the motile "corroding" strains into two groups . These groups correlated with the ultrastructural appearance of the cell walls . It is considered that isoelectric focusing techniques using 1 mm thick agarose gels may provide a more rapid method for ultimate differentiation of these organisms than other methods described so far.

J Toxicol Environ Health, 1984, 13(4-6), 545 - 51
Airborne endotoxins in a rice production commune in the People's Republic of China; Olenchock SA et al.; Concentrations of total and vertical elutriated airborne dusts were quantified for five work areas within a rice production commune near Shanghai, the People's Republic of China . Mean (+/- SE) commune levels of 15.69 +/- 1.91 mg/m3 and 6.00 +/- 1.47 mg/m3 were found for total and vertical elutriated dusts, respectively . Analysis of the airborne dusts for the presence of gram-negative bacterial endotoxins resulted in mean levels of 21.23 +/- 8.63 ng/mg total dust and 14.57 +/- 7.97 ng/mg elutriated dust, with respective airborne concentrations of 492.12 and 100.22 ng endotoxin/m3 . These data show that rice production in the People's Republic of China is associated with a dust burden that is contaminated with relatively high levels of endotoxins . The long-term respiratory effects of inhalation of these dusts should be studied.

Dev Biol Stand, 1984, 56, 371 - 83
Problems in the serodiagnosis of canine brucellosis: dog responses to cell wall and internal antigens of Brucella canis; Carmichael LE et al.; Three procedures are commonly used for the serodiagnosis of B . canis infection: 1) The rapid slide agglutination test (RSAT), 2) the tube agglutination test (TAT), and 3) the modified 2-mercaptoethanol tube agglutination test (2ME-TAT) . Hemocultures are always essential for diagnosis . The RSAT was developed to provide a presumptive diagnosis rapidly . It has been found accurate in identifying non-infected dogs; however false positive reactions are common due to shared determinants between the surface antigens of B . canis and certain other gram-negative bacteria . The RSAT has recently been modified to include brief reaction of test sera with 2ME (0.2M) prior to adding test antigen . The modification has improved specificity, but it has not eliminated false positive reactions . The TAT and 2ME-TAT are widely used . Although there is good agreement between tests, both suffer from lack of specificity but are valuable in kennels where B . canis has been identified by blood cultures . Agarose gel immunodiffusion (AGID) tests using extracted (SDC or hot PBS) cell wall antigenic complexes reveal precipitins in the sera of infected dogs usually 8 to 12 weeks postinfection (PI) that may persist 5 years . The antigenic complexes consist of at least 3 antigens; one (antigen '2R') appears to possess high B . canis specificity . Sera from noninfected dogs also may react nonspecifically in AGID tests that employ crude SDC or PBS antigenic extracts leading to 'false positive' interpretations . Cytoplasmic antigens gave up to four precipitin lines with sera from B . canis infected dogs . The antigens (protein or glycoprotein) were present in both S and R Brucella cells, but not in other gram-negative organisms examined . AGID tests that employed cytoplasmic antigens revealed precipitins against one or more (usually 2-3) antigens from PI months 4 through 64 . In some dogs, precipitins were present 12 months after the bacteremia had ceased, a time when other tests were diagnostically insignificant, or equivocal . No 'false positive' field sera reached with the cytoplasmic antigens.

Trans R Soc Trop Med Hyg, 1984, 78(4), 456 - 9
First report of septicaemic melioidosis in Hong Kong; So SY et al.; Septicaemia due to Pseudomonas pseudomallei was found in five patients; it complicated pneumonia in three . Four patients were old and had diabetes mellitus; the fifth patient, though young, was immunocompromised by steroid therapy . The clinical features were indistinguishable from those of other Gram-negative septicaemia . Four of the patients died . This was the first report of septicaemic melioidosis in Hong Kong where the disease might be endemic . Soil sampling and serological survey gave further supporting evidence . Early treatment using a bactericidal anti-Pseudomonal cephalosporin, such as ceftazidime, was suggested for this life-threatening infection.

Dev Biol Stand, 1984, 56, 199 - 211
Biochemical and biological properties of soluble protein preparations from Brucella abortus; Tabatabai LB et al.; Soluble salt-extractable protein antigens (CSP) from Brucella abortus may be of potential value as a vaccine and as a diagnostic reagent for the prevention and diagnosis of bovine brucellosis . These protein antigens, excluding protoplasmic proteins, are those present in the matrix of the outer membrane and in the periplasmic space of gram-negative bacteria . Our research deals mainly with the proteins of the periplasmic space and the nonporin proteins of the outer membrane . We have isolated a group of proteins from intact inactivated cells using mild, nondestructive extraction procedures . The proteins thus obtained range in molecular weight from 10 000 to 51 000 daltons as determined under denaturing conditions and from 10 000 to 124 000 daltons under nondenaturing conditions . The isoelectric pH of these proteins range from 3.5 to 10 with numerous bands in the pH 4.5 to 5.5 region, and a few bands in the pH 6 to 10 region . The low pH region also contains those protein bands which can be labeled with the membrane impermeant reagent, diazotized {125I}-iodosulfanilic acid . The soluble protein preparations are antigenic in guinea pigs, rabbits and cattle, and immunogenic in lemmings, guinea pigs and cattle . Cross-reacting protein antigens are present in protein preparations from B . abortus strains 19 and 2308 when tested with heterologous rabbit sera and with sera from vaccinated and experimentally infected cattle . The soluble protein antigens can be rendered more immunogenic by chemical modification of the primary amino groups with acyl anhydrides, especially with dodecanoyl anhydride . Immunization with dodecanoyl--modified proteins resulted in decreased humoral antibody levels (as measured in guinea pigs and cattle) without loss of protective activity, as measured in lemmings . Furthermore, the soluble proteins have proven to be a sensitive, stable and reliable reagent in the enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Brucella abortus in serum of vaccinated and experimentally infected cattle . Other soluble protein antigen preparations have also been evaluated and compared with strain 19 proteins for immunogenic properties and for use as an ELISA reagent.

J Clin Invest, 1984 Jan, 73(1), 160 - 70
Development and application of an enzyme-linked immunosorbent assay for the quantitation of alternative complement pathway activation in human serum; Mayes JT et al.; We have developed a new, specific, and highly sensitive enzyme-linked immunosorbent assay (ELISA) which quantitates activation of the alternative pathway in human serum, plasma, or on the surface of activators . The ELISA detects the third component of complement (C3b), proteolytic fragment of complement Factor B (Bb), and properdin (P) complex or its derivative product, C3b,P . In the method, activator-plasma mixtures, plasma containing an activated alternative pathway, or other samples are added to the wells of microtitration plates precoated with antibody to P . C3b, Bb,P or C3b,P complexes which become bound are quantitated by subsequently added, enzyme-labeled, anti-C3 . The resulting hydrolysis of the chromogenic substrate is expressed as nanograms of C3b by reference to a C3 standard curve . In addition to absolute specificity for activation of the pathway because of the nature of the complex detected by the assay, the ELISA is highly sensitive and able to reproducibly detect 10-20 ng/ml of C3b,P complexes in serum . This value corresponds to 0.0015% of the C3 in serum . In a series of studies to validate the parameters of the ELISA, reactivity was found to be dependent on the presence of alternative pathway proteins, the functional integrity of the pathway, and on the presence of magnesium . Sheep erythrocytes were converted to activators by treatment with neuraminidase . By using a variety of activators, the kinetics of activation and the numbers of bound C3b molecules quantitated by the ELISA were very similar to those measured by C3b deposition . The ELISA also detected identical activation kinetics when MgEGTA-serum and a mixture of the purified alternative pathway proteins were used as sources of the pathway . ELISA reaction kinetics also correlated with the restriction index, a measure of alternative pathway-activating ability . These studies cumulatively validate the ELISA as a direct and quantitative assay for alternative pathway activation . The sensitivity of the ELISA has permitted its use to detect direct alternative pathway activation by several viruses . The ELISA has also shown that certain classical pathway activators trigger the amplification loop of the alternative pathway while others do not . In addition, stable ELISA reactive complexes appeared in the supernatant of mixtures of serum with certain, but not other activators . The ability of the ELISA to detect activation which has already occurred and the stability of the reactive complexes permits studies of clinical sera . Normal human sera (20) contained low levels (5-20 ng/ml) of ELISA-reactive complexes . A proportion of sera from individuals with the adult respiratory distress syndrome (9-10), typhoid fever (8-10), malaria (3-5), gram-negative sepsis (9 of 47), acute trauma and shock (6 f 25), and systemic lupus erythematosus (3 of 29) showed elevated levels of complexes reactive in the alternative pathway ELISA . In contrast, nine sera from patients with circulating C3 nephritic factor were not reactive in the ELISA.

Scand J Urol Nephrol Suppl, 1984, 86, 87 - 91
Studies on rabbit hyperimmune, patient and blood donor serum with regard to bactericidal activity and serum antibodies against anaerobic curved rods from patients with bacterial vaginosis; Moi H et al.; Serum specimens from patients with bacterial vaginosis (BV) from whom anaerobic curved rods (ACR) were isolated, from healthy female and male blood donors and from rabbits immunized with ACR were tested for anti-ACR antibodies by indirect immunofluorescence and for bactericidal activity . The rabbit sera had antibody titres ranging from 1 : 160 to 1 : 2560 but no bactericidal activity could be demonstrated . Very low titres (1:10) of IgM antibodies were detected in serum from a few patients as well as from a few female and male blood donors, whereas no IgG or IgA antibody titres (less than 1:10) were demonstrated . No bactericidal activity was found in the human sera against ACR . We conclude that no systemic antibody response can be demonstrated in patients with BV from whom such organisms are isolated . Moreover, these organisms behave differently from other Gram-negative bacterial species, in that they are resistant to normal serum as well as the specific antibody-complement-mediated bactericidal activity.

Trans Assoc Am Physicians, 1984, 97, 172 - 81
Endotoxemia induced by antibiotic therapy: a mechanism for adrenal corticosteroid protection in gram-negative sepsis; Johnston CA et al.; The following have been demonstrated: With a standardized model of P . mirabilis peritonitis in ICR mice, when optimal doses of an aminoglycoside antibiotic (kanamycin) are given so as to reduce mortality maximally, abrupt and significant increments occur in endotoxemia, which cannot be attributed to impairment of clearance of endotoxin . Within the antibiotic-treated groups, these increments in endotoxemia correlate with mortality . The level of endotoxemia in the antibiotic-treated survivors is significantly greater than in animals dying without antibiotic therapy . MP, although unable to reduce mortality from P . mirabilis peritonitis in the absence of antibiotic therapy, does so when endotoxin is added to the challenge inoculum . It is concluded from these observations that aminoglycoside antibiotic treatment can shift the lethal mechanisms during P . mirabilis peritonitis from those involving bacterial proliferation and low levels of endotoxemia to those involving bacterial death and release of large amounts of endotoxin . It is postulated that this shift in lethal mechanisms could account for the ability of MP to reduce mortality when used in conjunction with antibiotics . Finally, since animals surviving after antibiotic treatment have significantly greater endotoxemia than those dying without such treatment and since MP does not reduce mortality in the absence of antibiotic therapy but does so if endotoxin is added to the challenge inoculum, it appears that endotoxin may not play a critical role in the pathogenesis of mortality from untreated P . mirabilis peritonitis . Whether the present correlation observed between the significant rises in endotoxemia produced by antibiotics and the protection afforded by MP extends to other models of gram-negative sepsis, or to human gram-negative sepsis, is under study.

Infection, 1984, 12 Suppl 1, S58 - 64
Central nervous system infections in the immunocompromised host; Armstrong D; Central nervous system (CNS) infections in the immunocompromised host are predictable according to the arm of the immune defense which is most defective . A limited number of organisms with a tendency to cause CNS infections will take advantage of the different types of immune defects . Signs and symptoms of CNS infection are often quite subtle in the immunocompromised host because of the diminished inflammatory response . Serologic responses may not be as reliable as in a normal host . Aggressive diagnostic approaches are often necessary, including early brain biopsy and abscess drainage . Aggressive therapy is often also necessary, including multiple potentially toxic antibiotics and devices to deliver them, such as indwelling intraventricular reservoirs . The role of the third generation cephalosporins is still not clear, and certainly they should not be used alone to treat aerobic gram-negative rod meningitis in neutropenic patients . Patients with the acquired immune deficiency syndrome (AIDS) have developed CNS infections with familiar agents producing familiar syndromes . The main CNS infection, however, is a subacute encephalitis and the causative agent remains undocumented.

Am J Pediatr Hematol Oncol, 1984 Fall, 6(3), 247 - 53
Supportive care for children with cancer . Guidelines of the Childrens Cancer Study Group . The role of granulocyte transfusions; Strauss RG; Review of the literature, as it pertains to the use of therapeutic and prophylactic granulocyte transfusions in the treatment of children with cancer, leads to the conclusion that granulocyte transfusions are of definite value in only one clinical setting . That setting is the severely neutropenic patient with Gram-negative septicemia who has failed to improve in response to appropriate antibiotics and in whom prompt bone marrow recovery seems unlikely . The efficacy of therapeutic granulocyte transfusions in other situations has not been demonstrated . Most investigators agree that prophylactic granulocyte transfusions should not be used because the benefits are few and the risks are great.

Dtsch Z Verdau Stoffwechselkr, 1984, 44(3), 109 - 17
{Endotoxins in gastroenterology--a medico-historical sketch}; Knoke M; First descriptions of effects of intestinal bacterial endotoxins date from the middle of the 19th century (P . L . Panum et al.) . The antitoxic function of the liver has been investigated by I . P . Pawlow in 1893 . At the turn of the last century the theory of "auto-intoxication" (C . Bouchard, I . I . Metschnikoff et al.) was well known, but there were also first systematic studies of the facultatively pathogenic intestinal bacteria (T . Escherich, H . Tissier, J . Strasburger) . In the twenties of our century V . van der Reis and L . Bogendoerfer worked out important fundamentals of human gastrointestinal microecology . Endotoxins as component of cellular wall of gram-negative bacteria are found by A . Boivin et al., J . W . Walker et al . First applicable proof for the detection of endotoxins was the pyrogen test with rabbits . The Limulus amoebocyte lysate test (J . Levin and F . B . Bang) has been employed as a more simple, rapid and sensitive method and was introduced in gastroenterology in a larger extent . Connections between endotoxinaemia and liver diseases, effects of endotoxins on gastrointestinal mucosa and on the course of shock are subjects of actual investigations.

Am J Ind Med, 1984, 6(2), 89 - 96
Sources of respiratory insult in the poultry processing industry; Lenhart SW et al.; Although the agricultural community has focused a great deal of attention on improving the overall health of poultry for production purposes, the health of poultry workers has, by comparison, received less direct attention . Recent studies suggest that poultry workers who come in direct contact with live birds may be at risk of pulmonary dysfunction, including the development of chronic respiratory diseases . Exposure of poultry workers to live birds results in potential pulmonary insult from a variety of allergenic and immunologic agents, as well as nuisance dust particles . In addition, marked levels of gram-negative bacterial endotoxins and antibiotic-resistant bacteria contaminate the occupational environment . Health care professionals should be aware of the potential sources of respiratory insult to workers in poultry processing and related industries.

Chir Pediatr, 1984, 25(1), 17 - 21
{Abdominal complications of ventriculo-peritoneal shunts in children . 65 cases}; Lortat-Jacob S et al.; From 1971 to 1982, among 1 200 children with hydrocephalus cured by ventriculo-peritoneal shunt, 65 have presented an abdominal complication . Some of them are rare and easily explainable: 2 intra-operative viscus perforations, 1 intestinal obstruction, 3 extrusions of the catheter into an inguinal hernia . Also rare, C.S.F . ascites (5 cases) et C.S.F . pseudocysts (3 cases) presented with typical symptomatology easily diagnosed, but their etiology is not clear . The main complications are peritonitis without perforation (32 cases) and migrations of the catheter in the colon (19 cases): their etiology is an intra-operative bacterial contamination, they happen during the following weeks or months after the shunt procedure . They are associated with 8 among 9 deaths of our data by gram negative germ meningitis . Removal of the peritoneal catheter and temporary external shunt are their main treatment.

Biol Neonate, 1984, 45(3), 119 - 24
Bacterial endotoxins in umbilical cord blood of neonates; Scheifele DW et al.; We tested umbilical cord blood from 255 infants for evidence of bacterial endotoxins . Using a Limulus lysate gelation technique, endotoxin-like activity (ELA) was detected in 13 (9.1%) of 142 term infants and in 23 (20.3%) of 113 preterm infants (p less than 0.05), in concentrations ranging from 30 to 3,000 pg Escherichia coli equivalent activity per milliliter of plasma . No factors predisposing to endotoxemia could be detected in term pregnancies but among ELA+ premature infants, placental cultures more frequently revealed gram-negative species (p less than 0.05) . Cord blood endotoxemia in the concentrations observed was without obvious consequence in mothers and term infants . Our data confirm the existence of natal endotoxemia, a potential mechanism of perinatal injury.

Mol Gen Genet, 1984, 194(1-2), 248 - 59
The role of insertions, deletions, and substitutions in the evolution of R6 related plasmids encoding aminoglycoside transferase ANT-(2"); Schmidt F; In 7% of gram-negative bacteria resistance to gentamicin is mainly mediated by plasmid-encoded aminoglycoside transferase ANT-(2") . The genome organization of 15 aadB plasmids (42-110 kb) was analyzed by restriction and hybridization techniques . They appeared to be IncFII-like replicons but were distinct from R6 by virtue of small substitutions in the transfer region . Aminoglycoside resistance genes aadB and aadA were located on Tn21 related elements . Only one of them was able to transpose its resistance genes mer sul aadA and aadB ( Tn4000 ), the other elements were naturally occurring defective transposons . In some of these structures deletions were identified at the termini, at sul, aadA , mer or transposition function--insertions adjacent to aadA or mer . The mode of these rearrangements and their site-specificity were considered with respect to the evolution of the Tn21 transposon family.

EMBO J, 1984 Jan, 3(1), 57 - 63
Analysis of copy number control elements in the region of the vegetative replication origin of the broad host range plasmid RK2; Thomas CM et al.; Broad host-range plasmid RK2 is able to replicate in a controlled manner in most Gram negative bacterial species . To analyze the elements of its control mechanism, we have measured the copy number in Escherichia coli of mini-RK2 replicons isogenic except for defined deletions in regions adjacent to the vegetative replication origin, oriVRK2, which have previously been implicated in copy number control because of their expression of plasmid incompatibility . The results indicate that while the previously defined 700-bp HaeII oriVRK2 fragment carries one copy control element (copA), a second (copB) lies at least partly outside this fragment towards the tetracycline resistance genes of RK2 . Deletions affecting both these regions give a mini replicon with a copy number of 35-40 compared with 4-7 for parental RK2 . Further incompatibility experiments indicate that targets for both incA (copA) and incB (copB) lie within the 700-bp HaeII oriVRK2 fragment.

J Immunol, 1984 Jan, 132(1), 369 - 75
A quantitative analysis of C3 binding to O-antigen capsule, lipopolysaccharide, and outer membrane protein of E . coli 0111B4; Joiner KA et al.; The binding of serum C3 to the O-antigen capsule (OAg Cap), lipopolysaccharide (LPS), and outer membrane proteins (OMP) of Escherichia coli 0111B4 was examined . Bacteria were intrinsically labeled with {3H} or {14C}galactose (*gal) in the OAg Cap and LPS moieties or with {14C}leucine (*leu) to label proteins . Organisms were then incubated in serum containing differentially labeled C3, the above fractions were separated, and the proportion of each binding to a column containing anti-C3 was measured . The OAg Cap fraction bound 72 to 82% of the C3, which bound to E . coli 0111B4 during incubation in absorbed 10% pooled normal human serum (10% PNHS) or absorbed 40% C8-deficient serum (C8D) . This distribution did not change when the organism was presensitized with immune IgG before serum incubation . A total of 2.93% +/- 0.48 of OAg Cap and 0.52% +/- 0.16 of LPS *gal bound specifically to Sepharose-containing antibodies to C3 (A:C3-Seph) after incubation in 10% PNHS; these values increased to 10.1% +/- 4.5 and 1.8% +/- 0.3, respectively, when C3 deposition was increased fourfold by incubation in 40% C8D . When encapsulated E . coli 0111B4 was incubated in 10% PNHS containing biotinylated C3, specific attachment of OAg Cap *gal to avidin-Sepharose was demonstrated in 1% sodium dodecyl sulfate (SDS), and complete release of bound *gal but not C3 occurred with 1 M NH2OH . When a mutant of E . coli 0111B4 lacking OAg Cap was incubated in 40% C8D, the outer membrane (OM) bound 85% of C3 . Five percent of OM *gal from the unencapsulated organism bound to A:C3-Seph in 0.05% SDS, indicating that the fraction of LPS molecules with bound C3 increased threefold in the absence of OAg Cap . OAg Cap does not contain protein, and no net specific binding of *leu from OAg Cap fractions to A:C3 was detectable; 2.4 to 3.6% of OM *leu bound to A:C3-Seph . Immunoprecipitation of 82.9% of OAg Cap *gal with antisera that were directed to E . coli 0111B4 was associated with co-precipitation of 69.5% of C3 in the capsular fraction . Therefore, the majority of C3 bound to E . coli 0111B4 was covalently attached to OAg Cap and LPS . As corroboration of experiments with whole bacteria, purified OAg Cap and purified LPS consumed C3 when incubated in serum in the fluid phase . These results are the first to evaluate the acceptor site for C3 deposition on a Gram-negative organism incubated in serum, and show that LPS, OAg Cap, and OMP are all major acceptor sites for C3 in nonimmune serum.

Antonie Van Leeuwenhoek, 1984, 50(5-6), 569 - 84
Excretion of proteins by gram-negative bacteria: export of bacteriocins and fimbrial proteins by Escherichia coli; Oudega B et al.; In gram-negative bacteria only few proteins are exported across both the cytoplasmic membrane and the outer membrane which forms an extra barrier for protein excretion . In this review we describe the mechanisms of production and export of two types of plasmid-encoded proteins in Escherichia coli . These proteins are the bacteriocin cloacin DF13 and the K88ab and K99 fimbrial subunits . Specific so-called helper proteins located at different positions in the cell envelope play an essential role in the export of these proteins . The genetic organization, subcellular location and functions of these helper proteins, as well as the effects of mutations and culture conditions on the export of the proteins are described . Models for the export mechanisms are presented and future application possibilities for engineering foreign protein excretion in E . coli with these export systems are discussed.

Acta Microbiol Pol, 1984, 33(3-4), 195 - 206
The cell wall of the obligate intracellular bacterial parasite of small free-living amoebae . I . Morphology and chemical composition of the rigid layer and peptidoglycan; Drozanski W et al.; The obligate intracellular bacterial parasite "OIBP" of small free-living amoebae, discovered by Drozanski (1956) was propagated in axenic culture of Acanthamoeba castellanii . The peptidoglycan prepared by chemical extraction of intact cells of the bacterium was examined in a transmission electron microscope and analysed chemically . Electron micrographs of heavy metal shadowed preparations revealed a bag-shaped membraneous structure resembling that of the peptidoglycan sacculi of Escherichia coli and the other gram-negative bacteria so far studied . The peptidoglycan may be present in a lipoprotein-peptidoglycan complex, as proteolytic enzyme treatment resulted in changes of the ultrastructure and in chemical composition . Results of chemical analysis of acid hydrolysed peptidoglycan indicate the presence of two aminosugars; glucosamine and muramic acid and also significant amounts of glycine together with three major amino acids; alanine, glutamic acid and diaminopimelic acid . It was shown that the peptidoglycan was, however, resistant to the hydrolytic action of egg-white lysozyme and to the lysosomal endo N-acetylmuramidases of amoebael origin.

Eur J Clin Microbiol, 1983 Dec, 2(6), 554 - 8
Treatment of gram-negative bacillary septicemia with cefoperazone; Lagast H et al.; Cefoperazone, a new broad-spectrum cephalosporin, was administered by continuous infusion or intermittent injection to 20 and 25 patients respectively with gram-negative bacillary septicemia most of whom had severe underlying disease . No difference was observed in the clinical response of the two groups . The overall rate of favourable response was 76% . Of the 40 non-neutropenic patients treated three died of infection despite sensitivity of the organism, three improved temporarily and three died as the result of superinfection . Three of the five neutropenic patients treated failed to respond to cefoperazone therapy, despite adequate serum bactericidal activity . Tolerance to cefoperazone was satisfactory . Emergence of cefoperazone-resistant strains leading to relapse or superinfection was not a major cause of failure.

J Virol, 1983 Dec, 48(3), 774 - 8
Accessibility of phosphatidylethanolamine in bacteriophage PM2 and in its gram-negative host; Brewer GJ et al.; The reaction of trinitrobenzenesulfonic acid with phosphatidylethanolamine in the cytoplasmic membrane of Alteromonas espejiana suggests that 50% of this lipid occupies the outer lamella . In PM2, similar analysis suggests that 56% of this lipid populates the outer lamella of the membrane, the surface of which accounts for 60% of the membrane area.

What Is Pcr?, What Is Anthrax?, What Is Amino Acid?, What Is Staphylococcus Aureus?, What Is Functional Genomics?, r, Bacterium, n, Bacteria, e, Bacteriology, a, Microbe, i, Microbes, r, Growth media, r, Microbiological, e, Escherichia coli, i, Antibiotics, o, Bacteria, a, Salmonella, o, Prokaryotes, i, Vibriosis, e, Bacteria, o, Gram positive, e, Kluyveromyces, e, E coli O157, c, S. cerevisiae, n, Pseudomonas, e, Streptococci, c, Escherichia coli, c, Staphylococcus, c, Enterobacters, o, S. cerevisiae, a, Staphylococcus, i, Eubacter

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005