J Am Vet Med Assoc, 1986 Oct 15, 189(8), 913 - 5 Serratia liquefaciens mastitis in a dairy herd; Bowman GL et al.; Serratia liquefaciens mastitis was detected and investigated in a 41-cow Holstein herd . Twenty cows were treated for mastitis over a 3-month period . Serratia liquefaciens was isolated from milk samples obtained from 8 of 12 cows tested during the epizootic . Results of an epidemiologic investigation suggested that extensive frostbite of the teats decreased the udder defense . Poor milking technique and hygiene were responsible for increased exposure of the damaged teats to potential udder pathogens . Treatment of each cow resulted in initial clinical improvement, but exacerbations occurred in 75% of the cows with documented S liquefaciens infections. Arch Phys Med Rehabil, 1986 Oct, 67(10), 722 - 5 Predicting renal calculus occurrence in spinal cord injury patients; DeVivo MJ et al.; This case-control study develops a model to predict the occurrence of renal calculi in patients with spinal cord injury (SCI) . Risk factors were assessed at the time of diagnosis in 25 patients who developed calculi, and at a comparable postinjury time period in 100 patients with SCI who remained calculus-free several years after injury . Logistic regression analysis was used to develop a predictive model; accuracy was assessed by using the model to classify all 125 patients studied . Renal calculi occurred more frequently on the right side and 72% of the affected patients developed a second calculus within two years . Patients who developed renal calculi were more likely to be older, have neurologically complete quadriplegia, have Klebsiella or Serratia infections, a history of bladder calculi, and high serum calcium values . The predictive model was 84% sensitive and 81% specific . While other determinants of renal calculi undoubtedly exist, these findings demonstrate that high risk patients may be identified with a comparatively small set of predictor variables . Although these findings are encouraging, use of any predictive model is meant only to supplement and not replace clinical judgement. South Med J, 1986 Oct, 79(10), 1252 - 5 Total hip arthroplasty: comparison of infection rates in a VA and a university hospital; Hofammann DY et al.; We reviewed charts of total hip arthroplasties from a ten-year period (January 1973 through December 1982) to determine the rate of infection at the Birmingham Veterans Administration Medical Center . Of 321 procedures, 296 were reviewed (92%) . Overall, there were 24 infections (8.1%), 13 of which (4.4%) were deep infections . Seven of the deep infections were due to Serratia marcescens, resulting in six implant removals . These figures are compared to the 1.1% deep infection rate after total hip arthroplasty at the adjoining University Hospital, University of Alabama at Birmingham . The main determinants for risk of infection at the VA Hospital were length of procedure and previous procedures on the affected joint. J Neurosurg, 1986 Oct, 65(4), 557 - 9 Brain abscess aspiration in nursery with ultrasound guidance . Case report; Nagle RC et al.; The authors report the case of a 1000-gm neonate who developed a frontal brain abscess due to a Serratia marcescens infection . The relationship of the mass to the neighboring ventricle and to the coronal suture was determined by computerized tomography . The mass was then successfully aspirated in the intensive care nursery with ultrasound guidance . A more prolonged operative procedure was avoided. J Infect Dis, 1986 Oct, 154(4), 670 - 5 Correlation of antibiotic synergy in vitro and in vivo: use of an animal model of neutropenic gram-negative sepsis; Chadwick EG et al.; The predictive value of in vitro studies of antibiotic interaction for clinical drug interactions is unclear . Five clinical isolates (two Klebsiella, two Pseudomonas aeruginosa, and one Serratia marcescens) were evaluated by the time-kill curve method for in vitro synergy between amikacin and imipenem . When we used the stringent definition of synergy of Hallander et al., no synergy was present for any study strain; however, when we used a more-conventional definition of synergy, these drugs interacted synergistically against all study strains . The results of these in vitro studies were correlated with in vivo interactions by using neutropenic infant rats injected ip with study organisms and given various treatment regimens . For 80% of the study strains, treatment of rats with amikacin and imipenem resulted in significantly greater survival than did therapy with either drug alone or than could be predicted by addition of survival rates achieved with either agent alone (P less than .005) . In vitro studies predicted this in vivo synergy in 80% of the cases when the more-conventional definition of synergy was used, whereas they were not predictive when the more-stringent definition of synergy was used . This rat model of neutropenia and gram-negative sepsis may provide more insight into in vivo drug interactions than do current methods. Arzneimittelforschung, 1986 Oct, 36(10), 1489 - 92 Protective effect of Nocardia rubra cell wall skeleton on experimental infection in normal and immunosuppressed mice; Mine Y et al.; The protective effect of Nocardia rubra cell wall skeleton (N-CWS) on experimental infections was investigated in normal and immunosuppressed mice . Pretreatment with N-CWS provided protection against acute systemic infections due to Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Serratia marcescens and Listeria monocytogenes in normal mice . N-CWS administered before or after challenge also showed protective activity against Herpes simplex virus infection in normal mice . N-CWS was the most effective against extracellular parasitic Pseudomonas aeruginosa infection when administered 1 day before challenge, but displayed the most potent protective activity against infection with Listeria monocytogenes, a facultative intracellular parasite, when administered 7 to 14 days before challenge . In mice with subcutaneous Pseudomonas aeruginosa infection, N-CWS suppressed abscess formation and decreased the viable cell count in the resultant abscess foci when administered either intraperitoneally or subcutaneously to the infected site . In addition, treatment with N-CWS markedly restored host defense ability against pseudomonal infection in mice immunosuppressed with cyclophosphamide, hydrocortisone and X-ray irradiation. J Biol Chem, 1986 Sep 25, 261(27), 12579 - 85 The von Willebrand factor-binding domain of platelet membrane glycoprotein Ib . Characterization by monoclonal antibodies and partial amino acid sequence analysis of proteolytic fragments; Handa M et al.; The glycoprotein Ib (GPIb), a two-chain integral platelet membrane protein, acts as a receptor for von Willebrand factor . In order to obtain information on the domain involved in this function, as well as on the structural organization of GPIb, the protein has been purified and submitted to limited proteolysis using three different enzymes . The resulting fragments were topographically oriented by means of partial NH2-terminal sequence analysis and immunological identification using monoclonal antibodies . One of these antibodies (LJ-Ib1) inhibited the von Willebrand factor-GPIb interaction completely, one (LJ-P3) partially, and one (LJ-Ib10) had no inhibitory effect . Three distinct fragments, the 38-kDa fragment produced by Serratia marcescens protease as well as the 45- and 35-kDa fragments produced by trypsin, had the same NH2 terminus as the intact GPIb alpha-chain (apparent molecular mass = 140 kDa) . These fragments and the alpha-chain reacted with the inhibitory antibodies . On the other hand, three fragments produced by Staphylococcus aureus V8 protease, one of 92 kDa similar to the previously described "macroglycopeptide" and two others of 52 and 45 kDa, had NH2-terminal sequences different from that of the GPIb alpha-chain and reacted only with the noninhibitor monoclonal antibody LJIb10 . Thus, the binding domain for von Willebrand factor resides near the NH2 terminus of the GPIb alpha-chain, whereas the carbohydrate-rich region is part of the innermost portion of GPIb and does not appear to be involved in the von Willebrand factor binding function. J Chromatogr, 1986 Sep 12, 364, 215 - 23 Ligand-exchange chromatography of nucleases; Varlamov VP et al.; The synthesis of chelating sorbents for ligand-exchange chromatography of enzymes is described . An inorganic support "Silochrom" and organic "Spheron", TSK-Gel HW 55 and cellulose were used as initial supports . The chelating sorbents contained iminodiacetic acid and iminodimethylphosphonic acid as stationary ligands . In order to obtain monofunctional sorbents, iminodiacetic acid was added in the form of its dimethyl ester . The concentration of stationary ligands on the sorbents varied from 10 to 100 mumol per ml sorbent . A chelating sorbent (in nickel form) was shown to be effective in the purification of exonuclease A5 from actinomyces . Electrophoretically homogeneous exonuclease A5 was obtained with a 25% yield . A chelating sorbent with iminodiacetic groups (in copper form) was applied to the isolation of endonuclease from Serratia marcescens directly from the culture medium . The capacity of the chelating sorbents for the endonuclease was studied as a function of the stationary ligand concentration . After one stage of purification, more than 70% pure enzyme was obtained with a yield exceeding 80%. J Gen Microbiol, 1986 Sep, 132 ( Pt 9), 2505 - 14 The role of surface polysaccharide in determining the resistance of Serratia marcescens to serum killing; Jessop HL et al.; Two O14:H12 strains of Serratia marcescens with different sensitivities to killing by normal pooled human serum were investigated . Complement binding, studied by measuring hydrophobicity and using rocket immunoelectrophoresis with anti-human C3, showed the sensitive cells (S1220) rapidly bound and fixed complement whereas the resistant cells (4444-60) bound less C3b . The strains had identical membrane protein composition . Crossed immunoelectrophoresis suggested that in S1220 cells the polysaccharide material including LPS was less antigenic and present in smaller amounts than in 4444-60 cells . This was confirmed by examining extracted polysaccharide material chemically and by SDS-PAGE . The resistant strain had 33% more phenol-extractable polysaccharide material than the sensitive strain, possibly comprising LPS with longer O antigen chain lengths, or a microcapsule of O antigen polysaccharide . Extra polysaccharide material on the surface of the resistant strain prevents complement components binding and reaching the hydrophobic membrane where lytic lesions occur. Antimicrob Agents Chemother, 1986 Sep, 30(3), 414 - 7 Novel morphological changes in gram-negative bacteria caused by combination of bulgecin and cefmenoxime; Nakao M et al.; The mode of action of bulgecin was investigated by examining its bactericidal and bacteriolytic activities, its effect on bacterial morphology, and its interaction with penicillin-binding proteins (PBPs) . Bulgecin alone did not show any antibacterial activity against Escherichia coli and Serratia marcescens, but in concert with cefmenoxime, it induced potent growth-inhibitory and bactericidal activities . Electron microscopic examination of E . coli cells exposed to bulgecin combined with cefmenoxime revealed that a bulge developed in the middle of the cell, and additional smaller bulges were formed halfway between the central bulge and the polar ends . At the site of bulge development, vesicular mesosomelike structures appeared in the cytoplasm, the peptidoglycan layer facing them became faint, and the outer membrane protruded to form blebs . These morphological changes were quite different from those caused by the mecillinam-cefmenoxime combination that produces big bulges in E . coli . When S . marcescens was exposed to the combination of bulgecin and cefmenoxime, not only bulge formation, but also branching of the cells was observed . Bulgecin neither showed affinity for any PBPs of E . coli nor affected the binding of cefmenoxime or mecillinam to the PBPs. Antimicrob Agents Chemother, 1986 Sep, 30(3), 398 - 402 Astromicin-induced membrane damage in Serratia marcescens; Umeda A et al.; The morphological changes in Serratia marcescens induced by astromicin were determined by a new technique of electron microscopy, a rapid freezing and substitution fixation technique, and a freeze-fracturing technique . Two structural changes were observed . One was damage to the cytoplasmic membrane, and the other was the accumulation of a large electron-dense mass in the cytoplasm . The damage observed in the cytoplasmic membrane was the disappearance of the unit membrane structure from the thin-sectioned profile of the drug-treated bacteria and the loss of the membrane particles from the fractured surface of the membrane . Damage to the membrane was also suggested by the results of examination of the spheroplasts for stability . The spheroplasts prepared from the drug-treated bacteria were unstable in an osmotically controlled buffer . Most of the spheroplasts were lysed within 3 h, whereas those prepared from control cells were stable for more than 15 h . The electron-dense mass in the cytoplasm was usually seen in the polar region of the cell in close contact with the cell membrane . These structural changes were not specific for astromicin but were also found in gentamicin-treated cells. J Med Microbiol, 1986 Sep, 22(2), 151 - 6 A survey of potential virulence factors in clinical and environmental isolates of Serratia marcescens; Franczek SP et al.; One hundred and forty seven isolates of Serratia marcescens were collected from diverse clinical and environmental sources in south-east Texas . Natural isolates were compared with hospital strains for the occurrence of 12 potential virulence determinants . Their overall frequency was as follows: haemolytic activity 48%; lecithinase 95%; lipase 95%; motility 99%; pigmentation 24%; plasmid carriage 46%; proteolytic activity 98%; siderophore activity 99%; urease activity 5%; mannose-sensitive haemagglutination 96%; mannose-resistant haemagglutination 61%; and mannose-resistant type-K haemagglutination (MR/K-HA) 68% . Clinical strains demonstrated a significantly higher occurrence of MR/K-HA (p less than 0.001) and non-pigmentation (p less than 0.01) than environmental isolates. Infect Immun, 1986 Sep, 53(3), 522 - 9 Degradation of protease inhibitors, immunoglobulins, and other serum proteins by Serratia protease and its toxicity to fibroblast in culture; Molla A et al.; We investigated the effect of the extracellular protease of Serratia marcescens on human serum constituents such as immunoglobulins, fibronectin, alpha 1-protease inhibitor, alpha 2-macroglobulin, lysozyme, and transferrin . At a very low concentration of Serratia 56-kilodalton protease (56K protease), purified human plasma fibronectin was degraded rapidly into three structural domains or small fragments . Immunoglobulin G3 (IgG3) and IgA1 were also degraded within 30 min with 1 microgram of this protease per ml, more rapidly than their other subclass of IgG or IgA . alpha 1-Protease inhibitor, which did not inhibit the 56K protease, was degraded similarly by the protease . These events were demonstrated by fluorescence polarization and sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The protease was considerably inhibited by human alpha 2-macroglobulin and chicken ovomacroglobulin . However, when there was a 2 M excess of ovomacroglobulin or a 4 M excess of alpha 2-macroglobulin over the 56K protease, about 25 or 40% proteolytic activity remained, respectively . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the protease degraded the alpha 2-macroglobulin extensively during prolonged incubation, which paralleled with regeneration of the protease activity . The protease also cleaved human lysozyme, although moderately . Human serum transferrin was degraded slightly, and human serum albumin was almost resistant to the 56K protease . The enzyme seemed to have no effect on reconstituted collagen, but it degraded rat tropocollagen and yielded fragments of beta and gamma chains by cleaving the intramolecular cross-links . Most of the above proteolysis by the 56K protease appears to result in a limited type of substrate specificity . Thus, the present study demonstrates that the protease is capable of degrading defense-oriented humoral proteins and tissue constituents . Furthermore, it is toxic to fibroblasts . These findings also clarified the possible role of Serratia protease as a virulence factor in the pathogenesis of serratial infections . We recently demonstrated this notion in vivo with rabbit cornea (R . Kamata et al., Ophthalmology 92:1452-1459, 1985). J Antimicrob Chemother, 1986 Aug, 18(2), 239 - 50 Enhanced bactericidal action of mouse macrophages by subinhibitory concentrations of monobactams; Iida-Tanaka K et al.; The effects of sub-minimum inhibitory concentrations (sub-MICs) of monobactams (aztreonam and AMA1080) on the host-parasite relationship were studied in an in vitro system using an established mouse macrophage cell line . The presence of sub-MICs aztreonam or AMA1080 enhanced significantly the macrophage bactericidal activity against Escherichia coli S615, Pseudomonas aeruginosa K1, Klebsiella pneumoniae 12 and Serratia marcescens US5 . Even four times the MIC of monobactams had no direct effect on macrophages . A synergistic bactericidal effect against E . coli was also observed with sub-MICs of monobactams and lysozyme or macrophage lysate . Furthermore, E . coli treated with sub-MICs of aztreonam was more sensitive to two bactericidal macrophage products, hydrogen peroxide and superoxide anion . These results suggest that the effects of monobactams are exerted on bacteria and not on macrophages; sub-inhibitory levels of monobactams may alter the bacterial cell rendering it more susceptible to bactericidal substances released by macrophages, thus favouring phagocytosis and killing by macrophages . Electron microscopic observations support these conclusions . This study provides evidence that monobactams at sub-MICs may work in partnership with host defenses against Gram-negative bacterial infections. Acta Ophthalmol (Copenh), 1986 Aug, 64(4), 456 - 62 Persistence of Serratia marcescens, Serratia liquefaciens and E . coli in solutions for contact lenses; Parment PA et al.; Twenty-four different brands of contact lens solutions were experimentally inoculated with strains of S . marcescens, S . liquefaciens and E . coli . Only flexol and hexidin could sufficiently suppress the growth of Serratia strains . If a soaking agent is to be effective in suppressing S . marcescens it must have a chlorhexidine concentration of at least 50 micrograms/ml and a thiomersal concentration of 10 micrograms/ml. Toxicology, 1986 Aug, 40(2), 199 - 205 Development of sensitive bacterial tests, exemplified by two mycotoxins; Lenz P et al.; For a sensitive bacterial test for mycotoxins the cup plate assay, based on growth inhibition, was optimised with Bacillus thuringiensis as test strain . Bacillus thuringiensis allowed the detection of 1.25 microgram kojic acid . A minimal amount of 12.5 micrograms kojic acid or 1.25 micrograms patulin was detectable by means of pigment suppression with isolated mutants of Serratia marcescens, whereas the wild type of this strain was insensitive. Nucleic Acids Res, 1986 Jul 25, 14(14), 5843 - 55 Cloning and sequencing of Serratia protease gene; Nakahama K et al.; The gene encoding an extracellular metalloproteinase from Serratia sp . E-15 has been cloned, and its complete nucleotide sequence determined . The amino acid sequence deduced from the nucleotide sequence reveals that the mature protein of the Serratia protease consists of 470 amino acids with a molecular weight of 50,632 . The G+C content of the coding region for the mature protein is 58%; this high G+C content is due to a marked preference for G+C bases at the third position of the codons . The gene codes for a short pro-peptide preceding the mature protein . The Serratia protease gene was expressed in Escherichia coli and Serratia marcescens; the former produced the Serratia protease in the cells and the latter in the culture medium . Three zinc ligands and an active site of the Serratia protease were predicted by comparing the structure of the enzyme with those of thermolysin and Bacillus subtilis neutral protease. Yale J Biol Med, 1986 Jul-Aug, 59(4), 453 - 9 Hospital-acquired gangrenous mucormycosis; Patterson JE et al.; A post-operative diabetic patient who had been treated for Serratia marcescens bacterial sepsis developed recurrent thrombosis of the left femoral artery following intra-arterial instrumentation . Pathological examination of arterial thrombus ultimately demonstrated invasive mucormycosis of the femoral artery and cultures of this material grew Rhizopus oryzae . The occurrence of cutaneous and subcutaneous mucormycosis is reviewed, as well as recently recognized nosocomial risk factors for mucormycosis, such as elasticized bandages and wound dressings. Antimicrob Agents Chemother, 1986 Jul, 30(1), 176 - 8 Frequency of aminoglycoside 6'-N-acetyltransferase among Serratia species during increased use of amikacin in the hospital; Larson TA et al.; The incidence of tobramycin-resistant, gentamicin-susceptible Serratia species at the Minneapolis Veterans Administration Medical Center decreased from an average 42.1 to 2.5% (P less than 0.001) during a 4.5-year period despite the predominant use of amikacin . These organisms were shown to express a 6'-N-acetyltransferase-modifying enzyme (EC 2.3.1.82) . Resistance was not shown to be plasmid mediated. J Pharm Pharmacol, 1986 Jul, 38(7), 510 - 4 Electrokinetic properties of endotoxins and their significance for the limulus amoebocyte lysate test; Baggerman C et al.; The electrokinetic properties of the endotoxins of Escherichia coli and Serratia marcescens have been examined . Both endotoxins are negatively charged, with the zetapotential being increased by the presence of cations whose relative influence resembles the Schulze-Hardy rule for colloid stability . Fe3+ and Th4+ ions are capable of reversing the negative charge of the endotoxin particles to positive . These cations were found to have a strong inhibitory effect on the activity of endotoxins in the limulus amoebocyte lysate test but the inhibitory effect did not parallel changes in the zetapotential because the effect occurred at concentrations too low to affect this parameter. J Hosp Infect, 1986 Jul, 8(1), 86 - 95 A comparison of strains of Serratia marcescens isolated from neonates with strains isolated from sporadic and epidemic infections in adults; Gaston MA et al.; As a result of the increased number of outbreaks of Serratia marcescens in special care baby units in the UK, a study was undertaken to compare strains isolated from outbreaks of neonatal infection with strains isolated from outbreaks of infections in adults and with isolates from sporadic infections . None of the biochemical, serological, bacteriological markers examined could distinguish the three groups of strains . When considered as groups there was no difference in the ability of the strains to survive desiccation on hands . Strains from neonatal and sporadic infections were more sensitive to antibiotics than adult epidemic strains . The feature common to all of the neonatal strains tested was the ability to agglutinate one or more species of erythrocytes in the presence of mannose . Only one strain in each of the other two groups possessed mannose resistant haemagglutinins. Am J Med, 1986 Jun 30, 80(6B), 22 - 8 Resistance surveillance programs and the incidence of gram-negative bacillary resistance to amikacin from 1967 to 1985; Gerding DN et al.; Data relating to amikacin resistance among gram-negative bacilli were obtained by means of a review of published literature and resistance surveillance studies . Data from the first several years of amikacin use are difficult to interpret because the 10-micrograms disk used for Kirby-Bauer susceptibility testing resulted in apparent greater resistance than the present 30-micrograms disk . A large United States susceptibility surveillance program that monitors antibiotic use has shown a trend since 1977 of greater susceptibility of Serratia species and greater resistance among Pseudomonas aeruginosa for all the aminoglycosides . Pseudomonas resistance to amikacin has shown the smallest increase of any aminoglycoside . Several hospitals (Strong Memorial Hospital, University of Maryland Cancer Center, and Minneapolis Veterans Administration Medical Center) have reported either no significant change or a decrease in resistance to amikacin when it was the most frequently used aminoglycoside . In a large, 14-center, prospective study, high-level use of amikacin resulted in a significant decrease in resistance to gentamicin and tobramycin (p less than 0.01) and a marginal increase (p less than 0.05) in amikacin resistance . Significantly increased amikacin resistance has been reported from two institutions, neither of which used amikacin as the predominant aminoglycoside . Overall, the high-level use of amikacin in large multi-center surveillance programs for as long as five years has not resulted in a significant increase in amikacin resistance rates at any of the individual institutions surveyed. J Biol Chem, 1986 Jun 15, 261(17), 7663 - 8 Ribokinase from Escherichia coli K12 . Nucleotide sequence and overexpression of the rbsK gene and purification of ribokinase; Hope JN et al.; The nucleotide sequence of a 1455-base pair TaqI-HinfI fragment of the rbs operon of Escherichia coli K12 has been determined . It includes the 3' terminus of rbsB (the gene for ribose-binding protein) and the entire rbsK gene, encoding ribokinase . Potential consensus promoter sequences and a stable stem-loop structure are present in the rbsB-rbsK intercistronic region . The regulatory significance of these sequence features is discussed with respect to the rbs operon . rbsK has been cloned downstream from the Serratia marcescens trp promoter on a multicopy plasmid . Cells harboring this plasmid, when grown on minimal ribose plus ampicillin, express ribokinase at the level of 2% of the soluble protein, and induction with indoleacrylic acid raises ribokinase levels another 8-fold . Ribokinase has been purified to homogeneity (216 mumol/min/mg) from a strain harboring this plasmid . Protein sequence analyses of peptides generated by cyanogen bromide cleavage and o-iodosobenzoic acid cleavage confirmed the translation initiation site and the reading frame of the DNA sequence . Amino acid compositions of native ribokinase and the C-terminal dodecapeptide agree with the predicted amino acid compositions, confirming the accuracy of the DNA sequence and the translation termination site. Invest Ophthalmol Vis Sci, 1986 Jun, 27(6), 932 - 9 Immunization against experimental Pseudomonas aeruginosa and Serratia marcescens keratitis . Vaccination with lipopolysaccharide endotoxins and proteases; Kreger AS et al.; Rabbits vaccinated with lipopolysaccharide endotoxins or with purified protease preparations from Pseudomonas aeruginosa and Serratia marcescens before corneal challenge with the viable bacteria exhibited significantly less corneal damage than rabbits not vaccinated with the bacterial products . However, the rabbits vaccinated with the lipopolysaccharide endotoxin preparations were significantly better protected than rabbits vaccinated with the bacterial proteases . Rabbits vaccinated with antisera raised against the proteases showed significantly less corneal damage than rabbits vaccinated with normal rabbit serum, and the passive protection was not significantly different than that elicited by active immunization against the bacterial proteases . The ability of the antiserum raised against the pseudomonas elastolytic protease to passively protect against severe corneal damage produced by experimentally induced pseudomonas keratitis was confirmed in mice . These findings support the idea that the bacterial endotoxins and proteases are virulence factors during the development of pseudomonas and serratia keratitis. J Bacteriol, 1986 Jun, 166(3), 937 - 44 Specific excretion of Serratia marcescens protease through the outer membrane of Escherichia coli; Yanagida N et al.; A DNA fragment of Serratia marcescens directing an extracellular serine protease (Mr, 41,000) was cloned in Escherichia coli . The cloned fragment caused specific excretion of the protease into the extracellular medium through the outer membrane of E . coli host cells in parallel with their growth . No excretion of the periplasmic enzymes of host cells occurred . The cloned fragment contained a single open reading frame of 3,135 base pairs coding a protein of 1,045 amino acids (Mr 112,000) . Comparison of the 5' nucleotide sequence with the N-terminal amino acid sequence of the protease indicated the presence of a typical signal sequence . The C-terminal amino acid of the enzyme was found at position 408, as deduced from the nucleotide sequence . Artificial frameshift mutations introduced into the coding sequence for the assumed distal polypeptide after the C terminus of the protease caused complete loss of the enzyme production . It was concluded that the Serratia serine protease is produced as a 112-kilodalton proenzyme and that its N-terminal signal peptide and a large C-terminal part are processed to cause excretion of the mature protease through the outer membrane of E . coli cells. Antimicrob Agents Chemother, 1986 Jun, 29(6), 1098 - 100 Bactericidal activity of ciprofloxacin against amikacin- and cefotaxime-resistant gram-negative bacilli and methicillin-resistant staphylococci; Simberkoff MS et al.; The MICs and MBCs of ciprofloxacin were determined for clinical isolates of antibiotic-resistant aerobic bacteria . Decreased susceptibility to ciprofloxacin of cefotaxime- and amikacin-resistant Serratia marcescens and amikacin-resistant Pseudomonas aeruginosa strains were noted . The data suggest that ciprofloxacin susceptibility should be carefully monitored in treating patients with hospital-acquired bacterial infections. Eur J Biochem, 1986 May 2, 156(3), 597 - 601 Structure of the O-specific polysaccharide from the lipopolysaccharide of Serratia marcescens O8; Oxley D et al.; Structural studies have been carried out on the O-specific polysaccharide from the lipopolysaccharide of the reference strain (CDC 1604-55) for serogroup O8 of Serratia marcescens . The polymer has a branched, tetrasaccharide repeating unit of D-galactose(Gal),D-glucose(Glc), and 2-acetamido-2-deoxy-D-glucose(GlcNAc) with the following structure: (Formula: see text) . The anomeric configuration assigned to the glucose residue differs from that (beta) previously proposed {Tarcsay, L., Wang, C . S., Li, S.-C . and Alaupovic, P . (1973) Biochemistry 12, 1948-1955} . The structure of the O8 polymer is identical with that of one of two polymers present in the cell envelope of a strain (CDC 1783-57) of S . marcescens O14. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1986 May, 182(3), 333 - 5 {Determination of alkylation of bacterial DNA as a rapid test for toxicological evaluation of alkylating xenobiotic agents}; Botzenhart K et al.; Alkylated purine bases from hydrolized DNA can be separated by HPLC and quantified with a fluorescence detector . We applied this method to bacterial DNA . 7-methylguanine was detected after treatment of Serratia marcescens with iodoacetamide, dimethyl sulfate and with polluted air. J Immunol Methods, 1986 Apr 17, 88(2), 175 - 83 A new simple fluorometric assay for phagocytosis; Oda T et al.; A highly sensitive, but simple and quantitative, fluorometric assay method for phagocytosis by cells such as macrophages and polymorphonuclear leukocytes was developed by utilizing fluorescent particles . Escherichia coli, Serratia marcescens, yeast, and latex particles were conjugated with fluorescein isothiocyanate and used as fluorescent particles . The assay procedure requires phagocytic cells, appropriate medium, fluorescent particles, sodium dodecyl sulfate, microtiter culture plate (24 wells), clinical centrifuge, and fluorescence spectrophotometer . One hundred assays can be done within 30 min after the incubation period . A time course analysis with this method showed that the phagocytosis of all these particles was dependent on temperature, and that the number of particles ingested by cells increased rapidly during the initial 30 min of incubation at 37 degrees C . Free fluorescent particles can be removed effectively by aspiration from the well . At 0 degree C, very few particles were ingested by cells or adsorbed onto the phagocytic cell surface as confirmed by fluorescence microscopy . An inhibitory effect of cepharanthin and sodium azide on phagocytosis was also confirmed by this method . The differential susceptibility of E . coli B and S . marcescens to phagocytosis also could be determined by this method. Z Kinderchir, 1986 Apr, 41(2), 78 - 80 Bacterial colonisation of the upper pouch in neonates with oesophageal atresia; Leung TS et al.; The bacterial flora in the upper oesophageal pouch of forty neonates with oesophageal atresia was studied at daily intervals preoperatively . Of the twenty-nine infants whose oesophagus was anastomosed within 24 hours of admission, no organisms were isolated in sixteen, despite the fact that only nine of these patients had antibiotics . The remaining thirteen grew oropharyngeal organisms . Of eleven infants having delayed anastomosis eight received antibiotics . All eleven grew organisms in the upper pouch . Pseudomonas and serratia grew only in those receiving antibiotics . These results suggest that prophylactic antibiotics are rarely indicated . Efficient continuous aspiration of the pouch is probably more important. Zh Mikrobiol Epidemiol Immunobiol, 1986 Apr, (4), 3 - 6 {New R-plasmids in strains of Serratia marcescens with multiple drug resistance}; Kurnosova LM et al.; In 4 S . marcescens polyresistant strains isolated from patients conjugative plasmids transferred to Escherichia coli have been detected . Two of these strains carry each one plasmid which codes resistance to 10 different antibiotics, including aminoglycosides which rarely occur in our country, and belongs to group IncC . The third strain is the host of 2 plasmids . One of them is similar to the above-mentioned 2 plasmids with respect to the incompatibility group and a set of markers, but additionally codes resistance to cephalosporins; the second plasmid has been determined as belonging to group IncM, unstable and capable of rendering the cells highly resistant only to aminoglycosides . And, finally, the fourth strain also carries 2 plasmids: one of them is unstable and belongs, supposedly, to group IncI alpha, and the second plasmid is stable and belongs to group IncM . The plasmid of group IncI alpha differs from all other plasmids of our Serratia by its capacity of rendering the cells highly resistant to chloramphenicol. Ann Thorac Surg, 1986 Apr, 41(4), 401 - 6 Airborne contamination during cardiopulmonary bypass: the role of cardiotomy suction; van Oeveren W et al.; Airborne contamination of the wound area and the cardiopulmonary bypass circuit during sham open-heart operations on dogs was studied . The air of the operating room (OR) was contaminated with two typeable bacterial strains . It was found that the number of wounds, blood specimens, oxygenators, and cardiotomy reservoirs contaminated with Staphylococcus aureus was related to the number of S . aureus present in the air of the OR, but that contamination with Serratia marcescens was related to the type of suction used . This form of contamination was considerably higher when air was aspirated together with blood into the suction line (p less than 0.05) . The oxygenator and cardiotomy reservoir were contaminated mainly by aspirating wound fluid from the airborne-contaminated wound area . The low number of sample sites positive for S . marcescens may be due to a better preserved host defense mechanism if only wound fluid is sucked . A rather high incidence of postoperative infections occurred even in dogs operated on in an OR with a low level of airborne contamination. Am J Med, 1986 Apr, 80(4), 753 - 4 Serratia marcescens osteomyelitis of the clavicle and sternoclavicular arthritis complicating infected indwelling subclavian vein catheter; Watanakunakorn C; Serratia marcescens bacteremia, osteomyelitis of the right clavicle, and septic arthritis of the right sternoclavicular joint developed in a 69-year-old woman after a right subclavian vein catheter was in place for five days . The infections were cured with the combination of gentamicin and ceftizoxime . There have been three previously reported cases of osteomyelitis of the clavicle following indwelling subclavian vein catheterization; two were caused by Staphylococcus aureus and one by Pseudomonas aeruginosa. J Bacteriol, 1986 Apr, 166(1), 244 - 52 Nucleotide sequence of the Escherichia coli motB gene and site-limited incorporation of its product into the cytoplasmic membrane; Stader J et al.; The motB gene product of Escherichia coli is an integral membrane protein required for rotation of the flagellar motor . We have determined the nucleotide sequence of the motB region and find that it contains an open reading frame of 924 nucleotides which we ascribe to the motB gene . The predicted amino acid sequence of the gene product is 308 residues long and indicates an amphipathic protein with one major hydrophobic region, about 22 residues long, near the N terminus . There is no consensus signal sequence . We postulate that the protein has a short N-terminal region in the cytoplasm, an anchoring region in the membrane consisting of two spanning segments, and a large cytoplasmic C-terminal domain . By placing motB under control of the tryptophan operon promoter of Serratia marcescens, we have succeeded in overproducing the MotB protein . Under these conditions, the majority of MotB was found in the cytoplasm, indicating that the membrane has a limited capacity to incorporate the protein . We conclude that insertion of MotB into the membrane requires the presence of other more hydrophobic components, possibly including the MotA protein or other components of the flagellar motor . The results further reinforce the concept that the total flagellar motor consists of more than just the basal body. Infect Immun, 1986 Mar, 51(3), 932 - 5 Cell surface hydrophobicity of pigmented and nonpigmented clinical Serratia marcescens strains; Rosenberg M et al.; The cell surface hydrophobicity of 10 pigmented and 4 nonpigmented clinical Serratia marcescens strains was studied, based on the ability of the strains to adhere to hydrocarbons and to polystyrene . The cell surface hydrophobicity depended greatly on growth temperature; all of the strains tested were adherent following growth at 30 degrees C, whereas none was adherent following growth at 38 degrees C . In previous studies, the pigment prodigiosin has been cited as responsible for cell surface hydrophobicity in various Serratia strains . However, the observed ability of the nonpigmented strains to adhere to the test hydrocarbons and to polystyrene indicates that Serratia strains can possess hydrophobic surface properties in the absence of this pigment . Moreover, strain 1785 cells were adherent whether they were grown at 30 or 36.5 degrees C, even though pigment was not synthesized at the higher temperature . In Escherichia coli correlations have been noted between increased cell surface hydrophobicity and the presence of mannose-specific adhesins; no such relationship was found in the S . marcescens strains tested . The expression of cell surface hydrophobicity in clinical S . marcescens strains at 30 degrees C and the loss of hydrophobicity at host temperatures raise the possibility that infective cells from the environment are initially hydrophobic, but lose this property upon subsequent proliferation within a host. J Heart Transplant, 1986 Mar-Apr, 5(2), 171 - 2 Pericardiectomy for effusive constrictive pericarditis after heart transplantation; Copeland JG et al.; The following is a case report of an unusual complication after heart transplantation . The patient was a 37-year-old man who underwent heart transplantation because of idiopathic cardiomyopathy . His postoperative course was complicated by cardiac tamponade, coagulopathy, and chronic constrictive pericarditis . After transplantation, he underwent three subsequent open-chest procedures: the first for tamponade, the second for Serratia mediastinitis, and the third for a pericardiectomy for constrictive pericarditis . Although constrictive pericarditis and pericardiectomy have been described following coronary bypass surgery and valve replacement, they have not yet been reported in a heart transplant recipient. J Antimicrob Chemother, 1986 Mar, 17(3), 327 - 32 Effects of ethylenediaminetetraacetic acid and gentamicin on the antibacterial activity of pyridone carboxylic acid derivatives against gram-negative bacilli; Miyake Y et al.; Ofloxacin (9-fluoro-3-methyl-10-(4-methyl-1-piperazynyl)-7-oxo-2,3-dihydro-7 H-pyrido-(1,2,3-de)1,4-benzoxazine-6-carboxylic acid) and enoxacin (1-ethyl-6-fluoro-1,4-dehydro-4-oxo-7-(1-piperazinyl)-1, 8-naphthyridine-3-carboxylic acid) are newly developed pyridone carboxylic acid derivatives with broad and potent antibacterial activities against Gram-negative and Gram-positive bacteria . Antibacterial activities of six pyridone carboxylic acid derivatives, including these two new antibiotics, were examined against Gram-negative bacilli in the presence and absence of ethylenediaminetetraacetic acid (EDTA) or gentamicin . The minimal inhibitory concentrations (MICs) of nalidixic acid, cinoxacin and piromidic acid were reduced by the addition of EDTA or gentamicin . However, the MICs of pipemidic acid, ofloxacin and enoxacin were unaffected . These findings indicated the high permeability of pipemidic acid, ofloxacin and enoxacin through the outer membrane . The effects of EDTA and gentamicin against Serratia marcescens were different from those against other Gram-negative bacilli. J Hosp Infect, 1986 Mar, 7(2), 149 - 54 An outbreak of Serratia marcescens transmitted by contaminated breast pumps in a special care baby unit; Gransden WR et al.; Laboratory surveillance of clinical isolates for Serratia spp . revealed a sudden increase from babies in the Special Care Baby Unit (SCBU) . It was established that breast-milk pumps on the post-natal wards were being disinfected inadequately, resulting in contamination of milk and cross-infection within the SCBU . Thirty babies were colonized and no deaths were attributable to the organism . Rectal carriage by the babies was common and often prolonged . The outbreak was brought under control when the method of disinfection of the pumps was changed from soaking in hypochlorite solution to washing at 80 degrees C. Eur J Biochem, 1986 Feb 3, 154(3), 581 - 5 Expression of the polyoma middle-size T antigen in Escherichia coli; Palme K et al.; We constructed a plasmid encoding a hybrid protein, consisting of the N-terminal signal sequence of the major outer membrane lipoprotein (lpp) of Serratia marcescens joined to the polyoma middle-size T antigen (mT antigen) . The hybrid protein expressed under the control of a lpp-lac hybrid promoter was synthesized at levels up to 5% of newly synthesized protein and could be accumulated in Escherichia coli strains carrying the Cap R mutation . The mT antigen produced in E . coli was precipitated by polyoma antitumor serum, and by serum directed against a synthetic peptide corresponding to the C terminus of the authentic mT antigen . The protein was secreted into the periplasmic space, from which it could be released by osmotic shock . The bacterial mT antigen had no detectable associated protein kinase activity. J Lab Clin Med, 1986 Feb, 107(2), 136 - 40 Bacterial adherence to intravenous catheters and needles and its influence by cannula type and bacterial surface hydrophobicity; Ashkenazi S et al.; Bacterial adherence to intravenous (IV) catheters and needles (cannulas) was studied morphologically by scanning electron microscopy and determined quantitatively with radiolabeled bacteria . Electron micrographs showed that bacteria adhered well to IV cannulas with formation of microcolonies . The adherence process was studied quantitatively, as related to cannula composition and bacterial surface hydrophobicity . The adherence of the bacteria examined (per square centimeter) was lowest to siliconized steel needles, higher to Teflon catheters, and highest to polyethylene catheters . The results for Staphylococcus aureus were (9.9 +/- 0.9) X 10(5) bacteria/cm2 adhered to steel needles, (37.2 +/- 2.8) X 10(5) bacteria/cm2 to Teflon catheters, and (168.4 +/- 15.6) X 10(5) bacteria/cm2 to polyethylene catheters . Hydrophobic bacteria (S . aureus and Serratia marcescens), as determined by their adherence to liquid hydrocarbons, adhered better than less hydrophobic species (Escherichia coli) . The role of hydrophobicity was documented by showing that hydrophobic S . marcescens adhered to IV catheters 18- to 27-fold better than its less hydrophobic mutants . It is concluded that IV steel needles have an advantage over plastic cannulas regarding bacterial adherence in vitro, and inasmuch as infectious complications in vivo were indeed shown to be lower with IV needles, their usage should be preferred. Antimicrob Agents Chemother, 1986 Feb, 29(2), 355 - 8 In vitro activity of A-56619 and A56620, two new aryl-fluoroquinolone antimicrobial agents; Smith BR et al.; The in vitro antimicrobial activity of two new aryl-fluoroquinolone antibiotics, A-56619 and A-56620, was compared with those of norfloxacin and several other antibiotics against 448 bacterial isolates . A-56620 was the most active agent tested . The usual 90% MIC of A-56620 was less than or equal to 2 micrograms/ml, except for enterococci, gentamicin-resistant Serratia marcescens, and gentamicin-resistant Pseudomonas aeruginosa, for which the 90% MIC was 4 micrograms/ml . A-56619 and norfloxacin were generally severalfold less active than A-56620 . Cross resistance was observed between the quinolone antibiotics and other unrelated antibiotic classes. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1986 Feb, 261(1), 85 - 94 Intraphagocytic bactericidal activity of ofloxacin compared with that of aztreonam and ceftriaxone against Serratia marcescens; Traub WH et al.; Addition of phenylbutazone (2 mg/ml) to 55 vol % of fresh defibrinated human blood permitted leukocytic ingestion of serum-resistant Serratia marcescens bacteria, but blocked phagocytic killing activity . The group A (phage tail) bacteriocin bA+ 16 served to kill extraphagocytic test bacteria . At greater than or equal to 2 X MBC, the DNA gyrase inhibitor ofloxacin revealed potent intraphagocytic bactericidal activity against S . marcescens test bacteria (99% kill; 3 h observation period) which corresponded to that of the control drug rifampin (97% kill) . The monobactam aztreonam (11% kill) and the third generation cephalosporin ceftriaxone (14% kill) corresponded to cefotaxime (26% kill) in terms of suboptimal intraphagocytic activity . Ofloxacin and aztreonam yielded additive effects following combination of supra-(2 X MIC) and inhibitory (MIC), but not sub-inhibitory (0.5 X MIC) concentrations with 55 vol % of defibrinated human blood against S . marcescens and Escherichia coli control strain ATCC 25922; sub- and inhibitory concentrations of ceftriaxone yielded indifferent effects. J Biochem (Tokyo), 1986 Feb, 99(2), 357 - 64 Instability of an arginine-overproducing mutant of Serratia marcescens and its stabilization; Takagi T et al.; Arginine productivity of an arginine-producing mutant of Serratia marcescens decreased during successive batch culturing . The mutant grew more slowly than the parent strains in a minimal medium, and spontaneously produced derivatives that grew more rapidly than the mutant . A large majority of the derivatives required N-acetylglutamate or arginine for growth, due to lack of N-acetylglutamate synthase, the argA gene product . The argA1 allele carried by the mutant was found to be relatively unstable . While the mutation rate in a stable argA mutant allele was less than 1 X 10(-8) per cell per generation, that in the argA1 allele was 9 X 10(-7) . The instability of the arginine productivity, therefore, was owing to both a disadvantage of the mutant in growth and a high mutability in the argA1 allele . In addition to the auxotrophs, the unstable arginine-producing mutant spontaneously produced at low frequency stable arginine-producing derivatives; among them, AT428 formed N-acetylglutamate synthase with a reduced affinity for glutamic acid . The derivative showed restored capability for propagation, and stably produced a large amount of arginine in the presence of glutamic acid or fumaric acid . By transductional analysis, the derivative was found to have acquired in the argA allele an additional mutation leading to the reduced affinity independently of the original one leading to the feedback-resistant enzyme. Arch Ophthalmol, 1986 Jan, 104(1), 79 - 83 Contact lens-associated microbial keratitis; Ormerod LD et al.; During a 14-year period, 42 cases of microbial keratitis were associated with contact lens (CL) wear . Pseudomonas aeruginosa was isolated in 40% of the cases and Staphylococcus in 31%; Streptococcus pneumoniae, alpha-hemolytic Streptococcus, and Serratia marcescens were the next most commonly isolated pathogens . There was a single fungal corneal ulcer . Bandage CL use was associated with a high prevalence of infection with quasi-commensal organisms and with polymicrobial keratitis, a pattern of disease quite distinct from that induced by other types of CLs . Marked visual loss frequently occurred . There was a disturbing increase in the number of infections associated with extended-wear CLs (worn for either aphakia or myopia) over the last 18 months of the study. J Fr Ophtalmol, 1986, 9(4), 305 - 9 {Hydrophilic contact lenses and pathogenic microorganisms}; Simitzis-Le Flohic AM et al.; A first study was conducted on 243 hydrophilic contact lenses: 65 were macroscopically abnormal and 100 were infected with fungi . Moreover on Sabouraud's medium with chloramphenicol, 30 bacterial strains were isolated of which 25 Pseudomonas sp . (10 Ps . cepacia) and 1 Serratia liquefaciens . Then a second study was conducted on 107 among the 243 lens solutions: the 50 of the B trade mark were spoiled with 12 X 10(7) bacteria/ml and the 57 of the C trade mark with 5 X 10(7) bacteria/ml . This quantitative bacterial difference was confirmed with a qualitative one: from B trade-mark 6 Pseudomonas aeruginosa strains were isolated and from C only one . The authors emphasize the importance of Ps . aeruginosa in corneal ulcers associated with contact lenses, of lens solution composition, and of preservative usefulness. Pediatr Pathol, 1986, 6(2-3), 351 - 8 Autopsy findings of Serratia meningoencephalitis in infants; Ariel I et al.; Serratia meningoencephalitis is often a fatal disease that causes widespread destruction of brain tissue despite aggressive antibiotic treatment . The autopsy findings of 2 cases are described . In a case caused by S . liquefaciens, previously not reported as the causative organism of meningoencephalitis, suppurative meningitis, ventriculitis, vasculitis, and extensive necrotic process of the brain matter were found . In the other case, caused by S . marcescens, the findings were those of acute and subacute abscesses with hemorrhagic necrosis. Chemotherapy, 1986, 32(6), 530 - 6 Passive protection of NMRI mice with commercial, intravenously applicable human immunoglobulin IgG preparations against Serratia marcescens; Traub WH; Three commercial, intravenously applicable human immunoglobulin IgG preparations (Gamma-Venin, Polyglobin, and Sandoglobulin) revealed low-titered O-agglutinins against the majority of 22 O-antigen reference strains of Serratia marcescens The three IgG preparations passively protected NMRI mice against intraperitoneal challenge with 8 of 19 selected, serologically defined O-antigen reference strains and clinical, nosocomially significant isolates of S . marcescens. Blood Cells, 1986, 12(1), 167 - 77 Phagocytic capacity of cytokineplasts from human blood polymorphonuclear leukocytes; Malawista SE et al.; Cytokineplasts (CKP) are membrane-bounded anucleate cytoplasmic fragments, induced from polymorphonuclear leukocytes (PMN) by the brief application of heat; derived from the cortical cytoplasm that gathers at the leading front of migrating PMN; and endowed with many of the motile properties of the parent cell . In this study we examine their phagocytic capacity by quantitative methods . CKP ingest Staphylococcus aureus and Serratia marcescens somewhat less avidly than do the corresponding intact PMN, yet rather impressively when one considers how restricted a portion of the parent cell they represent . Under the conditions employed, CKP killed about half as many of the bacteria presented to them as did their parent PMN . Thus, despite a heat-associated loss of demonstrable respiratory burst oxidase activity and a paucity of cytoplasmic granules, the organelle-depleted CKP deals with bacteria in a way that mimics its parent PMN. Microbiol Immunol, 1986, 30(6), 509 - 19 Survey of modifying enzymes and plasmids in amikacin-resistant Serratia marcescens; Katoh-Kanno R et al.; Forty amikacin-resistant strains of Serratia marcescens isolated from four different hospitals (A, B, C, and D) were examined for modifying enzymes and plasmids . Twenty-one of the isolates produced acetyltransferase that modified amikacin . Eighteen of the 21 acetyltransferase-bearing isolates were from different inpatients in hospital A and the other three were from hospital C . Amikacin resistance was mediated by conjugative plasmid of 24 megadaltons in 15 of the 18 acetyltransferase-bearing isolates of hospital A and by nonconjugative plasmids, derivatives of the 24-megadalton plasmids, in the remaining three isolates of the same hospital . The 24-megadalton plasmid determined aminoglycoside acetyltransferase (6') IV . This plasmid-borne enzyme conferred amikacin resistance on S . marcescens but not on Escherichia coli K12 . The frequency of transfer of the 24-megadalton plasmid from the S . marcescens isolate to E . coli K12 by conjugation was approximately 10(-7) (transconjugants/donors) and was 0.1% of that between E . coli strains . In acetyltransferase-bearing isolates from hospital C, the enzyme was mediated by a nonconjugative plasmid in one case and could not be associated with a plasmid in the remaining two cases . Neither enzymes nor plasmids could be associated with amikacin resistance of the isolates of the other two hospitals. Chemotherapy, 1986, 32(2), 148 - 58 Antibacterial antagonism of beta-lactam antibiotics in experimental infections; Kasai K; In vitro, 5 micrograms/ml of cefoxitin induced the highest beta-lactamase activity in Serratia marcescens TMS22, and the drug at this optimal dose required 2 h to increase the enzyme activity . The increasing enzyme activity was found to decline rapidly after the enzyme inducer effect was lost . When antagonism of cefoxitin against another beta-lactam, cefotaxime, was examined in infected granuloma pouch of rats, cefoxitin antagonized the antibacterial activity of cefotaxime administered at 4 and 6 h after cefoxitin (cefoxitin levels in pouch exudate were around 5 micrograms/ml) . The antagonism of an enzyme inducer and another antibiotic may be prevented by administering the non-enzyme inducer before the enzyme inducer exerts its inducer effect or after the enzyme inducer level decreases to an ineffective one. Circ Shock, 1986, 18(3), 171 - 8 Effect on cardiopulmonary changes of gram-negative endotoxemia in sheep after type-specific, cross-reactive, and nonspecific immune stimulation; Girotti MJ et al.; The purpose of this study was to examine the effects of prior nonspecific immune stimulation (BCG), cross-reactive immunization (E coli J5 0111 whole cells {J5 WC}, and core glycolipid {J5 CGL}), and type-specific immunization (Serratia marcescens core glycolipid {SM CGL}) on the cardiopulmonary variables and white blood cell counts of awake, monitored sheep following IV Serratia marcescens endotoxin . Comparison of cardiac output, pulmonary artery pressure, pulmonary capillary wedge pressure, pulmonary vascular resistance, alveolar-arterial oxygen gradients, and total white blood cells lead us to conclude that type-specific immunization (SM CGL) most effectively ameliorates the changes of gram-negative endotoxemia contrasted to nonimmunization . Core glycolipid cross-reactive (J5 CGL) immunization was somewhat more effective than whole-cell cross-reactive (J5 WC) immunization in this regard . Nonspecific immune stimulation (BCG) was able only to significantly decrease the changes in pulmonary vascular resistance compared to nonimmunization. Infect Immun, 1986 Jan, 51(1), 286 - 93 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and monoclonal antibodies as tools for the subgrouping of Escherichia coli lipopolysaccharide O18 and O23 antigens; Pluschke G et al.; The lipopolysaccharide (LPS) of Escherichia coli O18 isolated from a wide variety of sources was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) . Four different LPS types, designated O18A, O18A1, O18B, and O18B1, were identified . Most O18 strains possess O18A, O18A1, or O18B LPS types, and these types are clonally associated . A reference test strain with the classical O18ab designation possessed O18B LPS, while two reference O18ac strains possessed O18A and O18A1 LPS, respectively . A panel of 15 anti-O18A B-cell hybridomas was isolated . Enzyme-linked immunosorbent assays revealed that some of the monoclonal antibodies produced by these cells recognize different epitopes . Four of these antibodies suffice to distinguish the four O18 types . Numerous strains whose LPS had been typed by SDS-PAGE were tested by agglutination with seven monoclonal antibodies whose specificities had been determined by enzyme-linked immunosorbent assays . The results indicated a perfect correlation between the two methods . Rabbit antisera raised against O18A bacteria agglutinated boiled bacteria of each of the O18 LPS types efficiently . The antisera were adsorbed with bacteria possessing each of the LPS types . The adsorbed sera only distinguished between two groups: O18A and O18A1 versus O18B and O18B1, as shown by agglutination assays and Western blotting . E . coli O4 and O23 and Serratia marcescens O8 antigens, which are reputed to cross-react with O18, were also analyzed . One O4, one O8, and four O23 strains were tested . All made an LPS which was distinguishable from O18 LPS types by SDS-PAGE . Each O23 strain synthesized a different LPS, and three of them synthesized only few short chains . Some of the monoclonal antibodies reacted with O4, O8, and O23A LPSs . The results are interpreted as indicating that numerous E . coli O serogroups will prove to be chemically heterogeneous and that future analyses of subgroup heterogeneity should be guided by results from SDS-PAGE and rely preferentially on monoclonal antibodies as opposed to rabbit hyperimmune sera. Crit Care Med, 1986 Jan, 14(1), 26 - 8 High risk of nosocomial infection in the pediatric critical care patient; Donowitz LG; During this one-year prospective study, 61 (13.7%) of 444 patients admitted to the pediatric ICU at the University of Virginia Hospital developed nosocomial infections . By comparison, general medical/surgical ward patients had an overall 4.8% risk of acquiring an infection during their hospital stay . Patients who had prolonged ICU stays and those on plastic surgery, neurosurgery, and pediatric surgery services were more likely to become infected . The four bloodstream pathogens isolated in five episodes of hospital-acquired bacteremia were Staphylococcus epidermidis, S . aureus, Escherichia coli, and Serratia liquifaciens. Curr Genet, 1986, 10(12), 909 - 13 Isolation of the DNA sequence coding indole-3-glycerol phosphate synthetase and phosphoribosylanthranilate isomerase of Schizophyllum commune; Munoz-Rivas AM et al.; A Schizophyllum gene library was made in plasmid pRK9 . Plasmids from this library were tested for their ability to complement several auxotrophic mutations of Escherichia coli . The goal was to isolate a Schizophyllum auxotrophic gene that could be used to transform a corresponding Schizophyllum auxotrophic mutant to prototrophy . Complementation was observed only for E . coli trpC indole 3-glycerol phosphate synthetase (IGPS) and phosphoribosyl-anthranilate isomerase (PRAI) mutations . Plasmids with a Schizophyllum sequence coding for both IGPS and PRAI activities were recovered from E . coli transformants . Expression of the Schizophyllum gene (TRP1) in E . coli is probably dependent on the Serratia marcescens promoter of plasmid pRK9 . The DNA sequence containing the Schizophyllum TRP1 gene was not obviously rearranged in cloning. Chemotherapy, 1986, 32(1), 31 - 6 Activity of fosfomycin and R-plasmid conferring fosfomycin resistance among some clinical bacteria isolates in Nigeria; Obaseiki-Ebor EE; The in vitro activity of fosfomycin (an antibiotic that has clinically not been widely used in Nigeria) against 516 clinical bacteria isolates and the screening for the presence of R plasmid conferring resistance to fosfomycin among the test bacteria isolates were determined . In the presence of added glucose-6-phosphate (25 micrograms/ml) to the growth medium, all the isolates were inhibited at fosfomycin minimum inhibitory concentrations (MIC) of less than or equal to 32-64 micrograms/ml . Without the glucose-6-phosphate, fosfomycin had MIC75, MIC70, MIC48, and MIC20 at 64 micrograms/ml against Escherichia coli, Pseudomonas aeruginosa, Klebsiella spp., and Serratia spp., respectively, while the rest of the strains maintained about the same susceptibility as in the presence of glucose-6-phosphate . An R plasmid of about 59 megadaltons in size, conferring resistance to streptomycin, ampicillin, carbenicillin, and fosfomycin, was obtained from a Serratia liquifacens isolated in an area where fosfomycin had not been clinically used. Ann N Y Acad Sci, 1986, 485, 387 - 95 Platelet activation by alpha-thrombin is a receptor-mediated event; Harmon JT et al.; Computer-assisted data analysis of binding isotherms (LIGAND) has shown that human platelets have binding sites for alpha-thrombin of high (Kd 0.3 nM), moderate (Kd 10 nM), and low affinities (Kd 3 microM) . Application of similar techniques has shown that TLCK-thrombin does not, whereas PPACK-thrombin does, bind to the high-affinity binding site accessible to alpha-thrombin, but that both bind to the moderate and low-affinity sites . Treatment of platelets with Serratia marcescens protease destroys the high-affinity site but does not affect moderate-affinity binding . In accordance with this model, both modified thrombins compete with alpha-thrombin for platelet activation at the moderate-affinity site, but only PPACK-thrombin competes at the high-affinity site . These results establish that platelet activation by either low or moderate concentrations of thrombin are receptor-mediated events and explain the paradox of the differential effects of TLCK-thrombin on the binding and activation of platelets by alpha-thrombin. Physiol Bohemoslov, 1986, 35(5), 473 - 80 Effect of the activation of macrophages on the course of regeneration of rat liver following partial hepatectomy; Simek J et al.; Stimulation of the Kupffer cells with E . coli endotoxin (the purified lipopolysaccharide) or with prodigiosan (a polysaccharide from Serratia marcescens) 24 h before partial hepatectomy (resection of 65-70% of the liver) stimulated and intensified the onset of liver regenerative activity (evaluated from changes in liver DNA synthesis, the H5 labelling index and the mitotic activity of the hepatocytes) . Liver DNA synthesis increased together with the dose of endotoxin (i.v., from 25 to 1000 micrograms/kg body weight) . If E . coli endotoxin was injected during or 3 h after partial hepatectomy, partial inhibition of liver DNA synthesis was observed . In mice stimulated with zymosan (a polysaccharide isolated from yeast), administered 5 days before performing partial hepatectomy, proliferation of the hepatocytes (evaluated from changes in the 3H labelling index and in the mitotic activity of the hepatocytes) was evaluated . The results confirm that proliferation is correlated to the state of reactivity of the Kupffer cells. Adv Exp Med Biol, 1986, 198 Pt B, 71 - 8 Enhancement of vascular permeability upon serratial infection: activation of Hageman factor--kallikrein--kinin cascade; Matsumoto K et al.; A zinc dependent serratial 56K protease caused enhancement of vascular permeability followed by edema formation when injected into the guinea pig peripheral cornea, the subconjunctival space, or the skin . Because this enhancement was not affected by antihistamine, involvement of the kinin-generating system in this permeability enhancement was investigated . The 56K protease induced permeability much greater extent than that by bradykinin on weight basis, and more so on molar basis . The phenomenon was inhibited by soybean trypsin inhibitor, a well known inhibitor of plasma kallikrein, and also by corn trypsin inhibitor, which is the best inhibitor of the activated Hageman factor . In vitro experiments using numbers of synthetic peptide substrates, the 56K protease exhibited a similar substrate specificity to that of plasma kallikrein . Kallikrein is a known endogenous activator of Hageman factor . The enhancement by 56K protease was greatly augmented by inhibition of kininase II with Glu-Trp-Arg-Pro-Gln-Ile-Pro-Pro-OH (SQ 20,881), suggesting generation of bradykinin . Thus, these results indicate that the enhancement of vascular permeability induced by the 56K protease is caused by an activation of Hageman factor by 56K protease followed by subsequent activation of cascade amplification, and resulted in kinin generation in vivo. Folia Microbiol (Praha), 1986, 31(2), 106 - 12 Production of methionine and glutamic acid from n-alkanes by Serratia marcescens var . kiliensis; Ghosh BB et al.; A hydrocarbon-utilizing Serratia marcescens var . kiliensis grew and accumulated methionine and glutamic acid in a synthetic medium with hydrocarbon as sole carbon source . n-Hexadecane and ammonium phosphate were found as the most suitable carbon and nitrogen sources, respectively . Optimum pH for growth and methionine production was 7.2, and that for glutamic acid accumulation was 7.4 . Yeast extract significantly stimulated growth and amino acid production and could be substituted by cyanocobalamine . Benzylpenicillin, Tween 80, SDS or EDTA did not increase amino acid production . Under optimal cultural conditions in the laboratory the organism produced 1.68 g of glutamic acid and 0.78 g of methionine per litre. Int Arch Allergy Appl Immunol, 1986, 80(2), 200 - 10 Are cross-reacting natural antibodies multispecific? Milgrom F, Swierczynska Z. Human natural antibodies to antigens of Escherichia coli and Serratia marcescens were studied for cross-reactivity . The organisms were grown on synthetic media and extracted at 100 degrees C . The extracts were precipitated three times at 71% ethanol concentration and redissolved at the desired concentration . These preparations were referred to as Escherichia antigen (Ea) and Serratia antigen (Sa) . They readily coated red blood cells (RBC), which then could be used for passive agglutination tests . Human serum selected for this study had strong agglutinins combining with both antigens (cross-reacting antibody) and rather weak agglutinins combining with Ea only or Sa only, a property that became obvious from absorption experiments . Absorption of the serum with RBC coated by Ea removed all activity for Ea and most of the activity for Sa, and the opposite effect was noted after absorption of the serum by RBC coated by Sa . The activity against both antigens could be recovered by elution of antibodies at 56 degrees C from RBC coated by either Ea or Sa . Significantly, inhibition of the serum or the eluate with soluble antigens gave strictly specific results in that Ea abolished only the reaction with RBC coated by Ea and did not influence the reaction of RBC coated by Sa, and the opposite result was obtained upon inhibition of the serum with Sa . These results strongly indicated to us that the cross-reacting antibodies under study were multispecific, i.e., had different antigen-reactive sites (haptophore groups) for Ea and Sa . Further evidence supporting this contention was obtained from a study in which the cross-reacting antibody was 'labelled' by a bacterial antigen . To this end, the tested serum was neutralized with Sa and then reacted with RBC coated by Ea . From these RBC, antibodies were eluted from which Sa was recovered . A 'mirror image' experiment was also conducted in which Ea was recovered from antibodies that were blocked by Ea and reacted with RBC coated by Sa. Carbohydr Res, 1985 Dec 15, 145(1), 81 - 7 Structural studies of a neutral polymeric fraction from the lipopolysaccharide of Serratia marcescens C.D.C . 1783-57 (O14:H9); Brigden CJ et al.; A "neutral" polymer of glucose, galactose, and 2-acetamido-2-deoxyglucose (molar ratios 1:1:2) has been isolated from the lipopolysaccharide of Serratia marcescens strain C.D.C . 1783-57 (O14:H9) . Degradative and spectroscopic studies established that the polysaccharide has a branched tetrasaccharide repeating-unit of the structure shown . The polymer was absent from other strains of serogroup O14 studied, but a polymer differing only in the configuration of the glucose residue has previously been isolated from a strain of S . marcescens O8 . The polymer from strain C.D.C . 1783-57 also shares structural features with the Escherichia coli O18 antigen, which is known to be serologically related to the S . marcescens O8 antigen . (Formula: see text). Jpn J Antibiot, 1985 Dec, 38(12), 3487 - 96 {Studies on the combination action of sisomicin, gentamicin and cefmenoxime, ceftizoxime, cefoperazone, cefotetan against Serratia marcescens and Pseudomonas aeruginosa}; Kubo M et al.; The combined actions of sisomicin (SISO), gentamicin (GM) and cefmenoxime (CMX), ceftizoxime (CZX), cefoperazone (CPZ), cefotetan (CTT) against S . marcescens IFO-3735 and P . aeruginosa IFO-12689 were investigated . The results were summarized as follows . The synergistic effect of the combinations of SISO-CMX, SISO-CZX, SISO-CPZ, SISO-CTT, GM-CMX, GM-CZX, GM-CPZ and GM-CTT using the checker board dilution method on S . marcescens IFO-3735 and P . aeruginosa IFO-12689 were found . Especially the minimum FIC index value of combination of SISO-CTT and GM-CTT were rather low as 0.14 and 0.13, respectively and these synergistic effect was remarkably strong on S . marcescens IFO-3735 . With the killing kinetic method, all combinations tested showed the synergistic effect in both bacteria. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1985 Dec, 181(3-5), 309 - 19 {Transmissible formaldehyde resistance in Serratia marcescens}; Kaulfers PM et al.; It was possible to isolate a strain of Serratia marcescens from fresh clinical bacterial isolates which was 4-6 times more resistant against formaldehyde than other strains . It was shown that the strain harbours two plasmids with molecular sizes of 58- and 90 Mdal . It was demonstrated by conjugation-, transformation- and plasmid-curing experiments that the formaldehyderesistance is plasmid mediated and transferable to E . coli . It was shown by labelling with 14C-formaldehyde that the resistant strains bind much more formaldehyde than the sensible strains. Scand J Dent Res, 1985 Dec, 93(6), 555 - 9 Study of titanium screws as retrograde fillings using bacteria and dye; Luomanen M et al.; The tightness of retrograde titanium screw fillings and retrograde amalgam fillings was compared in 17 human, single-rooted teeth using Serratia marcescens bacteria in vitro . The root canals were subjected to instrumentation and irrigation, after which 2 mm was cut off from the apical end . Eight of the teeth were sealed using retrograde titanium screw fillings and nine using retrograde amalgam fillings . The teeth were suspended by means of wires in test tubes, with the crowns upwards and the roots immersed in trypticase soy broth . Suspensions of Serratia marcescens bacteria were placed in the root canals, and samples from the broth were plated daily . The bacteria penetrated the apical titanium screw seals in 2 to 7 days, and the retrograde amalgam fillings readily on the first day . Thus, the titanium screws seemed to provide a tighter seal . Staining with India ink showed that penetration had occurred at the tooth-filling margin and that the instrumentation had not caused any fractures to the roots. J Mol Biol, 1985 Nov 5, 186(1), 87 - 96 Isolation and characterization of mutants affecting functional domains of ColE1 RNAI; Dooley TP et al.; The control of DNA replication initiation in the plasmid ColE1 is mediated by RNAI, a 108 nucleotide plasmid-encoded RNA that is entirely complementary to the 5'-terminal region of the replication primer RNA . RNAI acts in trans to inhibit primer maturation . Previously, we constructed a plasmid in which the ColE1 RNAI was separated from the primer and placed under transcriptional control of the Serratia marcesens tryptophan promoter . This plasmid provides RNAI in trans in vivo and mediates ColE1-type incompatibility . To determine the critical structural and functional domains of RNAI, we have undertaken a mutational analysis of the RNAI gene carried by this plasmid . We have selected mutants that no longer mediate ColE1-type incompatibility in trans . From the DNA sequences of 18 mutants we have identified mutations at nine new sites in RNAI . In addition, we have determined the secondary structural features of several mutant RNAI species and compared them to wild-type RNAI . Analysis of these mutations has revealed several key features of RNAI secondary structure and function . The domains of RNAI identified in this work which are essential for its function are: the single-stranded loop regions; the integrity of the double-stranded stems; and the single-stranded 5' terminus. Rev Infect Dis, 1985 Nov-Dec, 7 Suppl 4, S538 - 44 Gram-negative bacterial infections: a look at the past, a view of the present, and a glance at the future; Weinstein L; An overview of infections caused by gram-negative bacteria over the past 40 years discloses remarkable changes in their specific etiology and management . These organisms were responsible for disease in the preantibiotic era but at a much lower frequency than at present, and fatality rates were generally high . Bacteria such as Pseudomonas and Serratia, now relatively commonly involved in infection, rarely, if ever, caused disease before antibiotics became available . Although often present in surgical wounds, these organisms clearly were colonizers rather than pathogens . It is clear that the use of potent antimicrobial agents has been responsible for a general decrease in the fatality rates associated with gram-negative bacterial infections . However, it is evident that their use has been involved in the development of potentially lethal complications not seen in the past . Factors presently recognized as playing important roles in the problems associated with the treatment of gram-negative bacterial infections are the increasing number of older and sometimes debilitated patients and the longer survival of individuals with tumors or leukemia who, because of immunosuppression due to disease or treatment, are highly susceptible to invasion by almost any organism . Factors that have increased the magnitude of the problem of treatment of gram-negative bacterial infections include an increasing frequency of bacterial resistance to one or more antibiotics and a tendency of physicians to use more than one drug, sometimes as many as three or four, to treat gram-negative bacterial infections, especially in immunosuppressed people . Such polypharmacy is responsible for the development of suprainfections, some of which are caused by organisms very difficult to eradicate. Biol Reprod, 1985 Nov, 33(4), 925 - 33 Follicle stimulating hormone binding inhibitor produced by the bacteria Serratia interacts with receptor for follicle stimulating hormone in calf testis membranes; Sluss PM et al.; Bacteria of the genus Serratia, including a strain of Serratia liquifaciens isolated from contaminated porcine follicular fluid, produced an inhibitor of 125I-human follicle-stimulating hormone (hFSH) binding to calf testis membranes in vitro . In order to evaluate its potential usefulness and significance, we undertook studies to identify the site of action of this inhibitor . Large quantities (grams) of inhibitor-containing material (SL-1) were obtained by enrichment culture techniques and its chemical composition was determined . Follicle-stimulating hormone-binding inhibitory activity (FSH-BI) in SL-1 was associated with a protein-containing substance of approximately 30,000 Mr and also with larger Mr (greater than 300,000) forms . Preincubation studies demonstrated that binding inhibition by SL-1 was due to effects on the membranes rather than effects on the radioligand (125I-hFSH) . Kinetics studies indicated that FSH inhibition by SL-1 was a relatively slow process that reached steady-state conditions between 23 and 25 h at 20 degrees C, in contrast to FSH, which reached steady-state conditions by 12 h at 20 degrees C . Estimates of FSH-BI activity, e.g., mass required to produce a 50% inhibition of 125I-hFSH binding, varied drastically when these kinetics differences were not taken into account . Our observations emphasize the need to establish steady-state conditions for each ligand before assessing mechanisms of action using Michaelis-Menton assumptions (e.g., competitive binding assays, Scatchard analyses).(ABSTRACT TRUNCATED AT 250 WORDS) Infect Immun, 1985 Oct, 50(1), 218 - 24 Bacterial adherence to human endothelial cells in vitro; Ogawa SK et al.; Differences in the ability of bacteria to adhere to normal valvular endothelium may account for the predominance of particular species as pathogens in acute endocarditis . An in vitro adherence assay was developed to simulate the host surface encountered in acute bacterial endocarditis by using confluent monolayers of human endothelial cells . Adherence of 32 gram-positive and -negative blood culture isolates to this surface was compared . All five Staphylococcus aureus strains tested were highly adherent to endothelial cells, as was one gram-negative strain (Serratia marcescens) . The remaining gram-positive and -negative isolates, including four viridans streptococci, were relatively nonadherent . Transmission electron microscopy demonstrated attachment of Staphylococcus aureus and invagination of the underlying endothelial cell membrane at 1 h followed by engulfment of large numbers of bacteria after 3 h . The intracellular bacteria appeared to be contained within vacuoles . Preferential attachment of some strains of bacteria, in particular Staphylococcus aureus, to human endothelial cells occurred in vitro, suggesting that adherence is an important determinant of bacterial pathogenicity in acute endocarditis . Active uptake of bacteria by endothelial cells may help account for the virulence of Staphylococcus aureus in endovascular infections and for the ability of this organism to establish multiple metastatic foci of infection. Zh Mikrobiol Epidemiol Immunobiol, 1985 Oct, (10), 19 - 21 {Properties of Serratia marcescens strains isolated in septic diseases of newborn infants}; Abrikosova NIu et al.; As the result of the study of blood and liquor samples from 120 newborns, Serratia marcescens was isolated in 21 cases (17.5 %) . 8 strains were isolated from the environment of these patients . Almost all strains isolated from both the patients and the environment (with the exception of one environmental strain) belonged to serotype 04 . The isolated S . marcescens strains were resistant to penicillin, ampicillin, streptomycin, kanamycin, oxacillin, methicillin, ceporin and moderately sensitive to polymixin . 2 strains from the environment and 9 strains from the patients were mildly sensitive to gentamicin . In one hospital all isolated strains were found to have 2 transmissive R plasmids with the molecular weight 40 and 60 megadaltons . The presence of R plasmids with the same molecular weight in all S . marcescens strains isolated in this hospital, as well as their serological identity, suggest that in all patients infection originated from a common source. J Biochem (Tokyo), 1985 Oct, 98(4), 1139 - 42 Preliminary X-ray studies on Serratia protease; Katsuya Y et al.; Preliminary X-ray studies on Serratia protease have been carried out using crystallographic and small angle scattering techniques . The enzyme has been crystallized in three different crystalline forms by microdialysis and vapor diffusion methods using 50 mM phosphate buffer, pH 6.0, at 24 degrees C . They have orthorhombic space groups: C222(1) for one form and P2(1)2(1)2(1) for the other two forms . A small angle X-ray scattering study showed that the radius of gyration and the maximal dimension of the molecule in aqueous solution are 26.6 A and 94.5 A, respectively . The molecular weight of the enzyme was determined to be 45,000-48,000 by various physical methods. Ophthalmology, 1985 Oct, 92(10), 1452 - 9 The serratial 56K protease as a major pathogenic factor in serratial keratitis . Clinical and experimental study; Kamata R et al.; A possible cause and the difference in clinical severity of serratial keratitis were investigated . Two strains of Serratia marcescens were isolated: one from a patient with severe liquefactive keratitis, who had diabetes mellitus, and one from a patient with mild superficial keratitis, but who had no underlying disease . When the same numbers of bacteria were injected separately into corneas of the same rabbits or guinea pigs, the strain from the first patient elicited severe corneal destruction, remarkable intracorneal edema; and liquefactive necrosis, but the strain from the second caused mild keratitis with erosion or intracorneal abscess . The keratitis induced by the former strain required a longer time to heal, and the prognosis was poorer than that for the other keratitis . Therefore, the difference in severity between the two cases of experimentally induced keratitis paralleled that of the clinical cases . Thus, the severity of the serratial keratitis might be attributed more to the virulence of the bacteria than the condition of the host . The virulence factor seemed to be a heat-labile metabolic product (or products) of the bacteria . To clarify this virulence factor, the major secretory protease (56K protease) produced by these two strains of bacteria was compared by using in vitro and in vivo systems . The virulent strain produced about ten times more protease during culture than the less virulent strain . When injected into the corneas of experimental animals, the 56K protease from the virulent strain induced severe lesions similar to those caused by the living virulent strain of bacteria . These results indicated that one of the major factors causing the virulence was correlated with the tissue destructive 56K protease produced by S . marcescens. J Bacteriol, 1985 Oct, 164(1), 217 - 22 Leucine regulation of the ilvGEDA operon of Serratia marcescens by attenuation is modulated by a single leucine codon; Hsu JH et al.; The effect of leucine limitation and of restricted leucine tRNA charging on the expression of the ilvGEDA operon of Serratia marcescens was examined . In this organism, the ilv leader region specifies a putative peptide containing only a single leucine codon that could be involved in leucine-mediated control by attenuation (E . Harms, J.-H . Hsu, C . S . Subrahmanyam, and H . E . Umbarger, J . Bacteriol . 164:207-216, 1985) . A plasmid (pPU134) containing the DNA of the S . marcescens ilv control region and three of the associated structural genes was studied as a single chromosomal copy in an Escherichia coli strain auxotrophic for all three branched-chain amino acids . The S . marcescens ilv genes responded to a multivalent control similar to that found in other enteric organisms . Furthermore, the S . marcescens ilv genes were derepressed when the charging of leucine tRNA was restricted in a leuS derivative of E . coli that had been transformed with pPU134 . It was concluded that ribosome stalling leading to deattenuation is not dependent on either tandem or a consecutive series of codons for the regulatory amino acid . However, the fact that the single leucine codon is a less frequently used codon (CUA) may be important . The procedure for obtaining the cloned ilv genes in single chromosomal copy exploited the dependence of ColE1 replicons on the polA gene . The cloning experiments also revealed a branched-chain amino acid-glutamate transaminase in S . marcescens that is different from transaminase B. Obstet Gynecol, 1985 Sep, 66(3 Suppl), 48S - 51S Gonococcal endocarditis during pregnancy: simultaneous cesarean section and aortic valve surgery; Burstein H et al.; Gonococcal endocarditis is a rare and potentially fatal consequence of disseminated gonococcal infection . Presented is the first known case of culture-proved gonococcal and serratia endocarditis in pregnancy . The case was further complicated by fetal distress at 30 weeks' gestation as a result of maternal decompensation from worsening congestive heart failure secondary to rapid destruction of her aortic valve . Consequently, cardiopulmonary bypass with subsequent aortic valve replacement and implantation of a left ventriculoaortic shunt was initiated immediately after an emergency cesarean section. J Gen Microbiol, 1985 Sep, 131 ( Pt 9), 2343 - 8 Immunochemical characterization of the outer membrane complex of Serratia marcescens and identification of the antigens accessible to antibodies on the cell surface; Jessop HL et al.; Serratia marcescens New CDC O14:H12 contains major outer membrane proteins of 43.5 kDal, 42 kDal (the porins) and 38 kDal (the OmpA protein) which can be separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis . Immunoblotting of whole cell or outer membrane preparations using antiserum raised against the whole cells revealed similar complex patterns of antigens . The OmpA protein was the major immunogen, although six other outer membrane proteins were also detected; the porins reacted only weakly with antibodies in this system . Immunoabsorption of antisera with whole cells showed that only the O antigenic chains of lipopolysaccharide and the H (flagella) antigens were accessible to antibody on the cell surface . Failure to detect the OmpA protein and other envelope antigens in this way suggests that their antigenic sites are not able to react with antibodies and are possibly masked by the O antigen. Can J Microbiol, 1985 Aug, 31(8), 730 - 5 Effect of bacteria on the inactivation and adsorption on clay minerals of reovirus; Lipson SM et al.; This investigation studied the antiviral activity of, and the utilization of viruses as substrates by, bacteria . Reovirus type 3 and bacterial species representative of those endemic to sewage, aquatic, and terrestrial habitats were used in the model systems . Culture supernatants from Bacillus subtilis maintained for 5 days in a minimal salts medium displayed antiviral activity, but supernatants from Escherichia coli or Serratia marcescens did not . Both live and toluene-killed cells reduced the inactivation of reovirus during 4 days of incubation at 23 +/- 2 degrees C . This protective effect was more pronounced with killed than with live cells of B . subtilis, confirming the presence of an antiviral component(s) in this species and indicating that the component(s) was metabolic in origin . When reovirus was presented to these bacteria as a sole source of carbon, some growth (determined spectrophotometrically) of B . subtilis and S . marcescens occurred with reovirus concentrations of 3.1 X 10(6) and 8.2 X 10(6) mean tissue culture infective dose-fifty X mL-1, respectively . Growth of S . marcescens did not occur with a reovirus concentration of 8.0 X 10(4) mean tissue culture infective dose-fifty X mL-1, nor did that of E . coli with any virus concentration used in this study . Adsorption of reovirus on kaolinite was enhanced by the culture supernatant from S . marcescens and on montmorillonite, albeit to a lesser extent, by that from E . coli . The effect of culture supernatants from B . subtilis on the adsorption of reovirus on clay minerals could not be determined, as a result of the antiviral component produced by these cells . The virus was not adsorbed on the bacteria. J Antimicrob Chemother, 1985 Aug, 16(2), 227 - 34 Clinical efficacy of a synergistic combination of cefotaxime and amikacin against multiresistant Pseudomonas and Serratia infections; Maslow MJ et al.; The synergistic activity of cefotaxime and amikacin against 21 highly resistant Pseudomonas aeruginosa and Serratia marcescens isolates was evaluated in-vitro by the checkerboard tube dilution method and in-vivo in five patients with serious infections caused by these organisms . All isolates were resistant to gentamicin, tobramycin, amikacin, and cefotaxime . Synergy was observed in 90% of isolates and occurred when the MIC of amikacin was less than 256 mg/l and of cefotaxime less than 1024 mg/l . A clinical response occurred in 100% and bacteriological cure in 80% of patients . Our results demonstrate a high degree of synergism between amikacin and cefotaxime in-vitro and clinical efficacy in the treatment of infections due to multiply-resistant Pseudomonas and Serratia species. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1985 Aug, 260(1), 35 - 40 Fibrinolytic activity of purified Serratia marcescens metalloproteases; Traub WH et al.; Two purified metalloproteases of Serratia marcescens revealed fibrinolytic activity, i.e., degraded human fibrin in agar as well as commercially available human fibrinogen and fibrinogen in fresh human plasma. Appl Environ Microbiol, 1985 Aug, 50(2), 487 - 90 Droplet enrichment factors of pigmented and nonpigmented Serratia marcescens: possible selective function for prodigiosin; Burger SR et al.; Drops produced by bursting bubbles provide a mechanism for the water-to-air transfer and concentration of matter . Bacteria can adsorb to air bubbles rising through bacterial suspensions and enrich the drops formed by the bubbles upon breaking, creating atmospheric biosols which function in dispersal . This bacterial enrichment can be quantified as an enrichment factor (EF), calculated as the ratio of the concentration of bacteria in the drop to that of the bulk bacterial suspension . Bubbles were produced in suspensions of pigmented (prodigiosin-producing) and nonpigmented cultures of Serratia marcescens . EFs for pigmented cultures were greater than EFs for nonpigmented cells . Pigmented cells appeared hydrophobic based on their partitioning in two-phase systems of polyethylene glycol 6000 and dextran T500 . The surface hydrophobicity of pigmented cells may result from the hydrophobic nature of prodigiosin and could account for the greater ability of these bacteria to adsorb to air bubbles and enrich airborne droplets . Enhancement of the aerosolization of S . marcescens may be a selective function of the bacterial secondary metabolite prodigiosin. Am J Med, 1985 Jul 15, 79(1A), 61 - 7 Pneumonia caused by gram-negative bacilli; Karnad A et al.; Gram-negative bacillary pneumonia has become an increasingly important disease in immunosuppressed, elderly, and hospitalized patients . The clinical features, etiologic agents, population at risk, treatment, and outcome in patients with well-documented gram-negative pneumonia were compared in two groups of patients: those with bacteremic pneumonia and those with nonbacteremic pneumonia documented by transtracheal aspiration . Clinical features were frequently subtle in both groups . A wide range of gram-negative bacilli were implicated as pathogens and pneumonias documented by transtracheal aspiration were frequently mixed infections . Pseudomonas aeruginosa and Serratia marcescens were the most common pathogens causing bacteremic pneumonias, whereas Escherichia coli and Klebsiella were more common in the nonbacteremic group . Gram-negative bacillary pneumonia was frequently a lethal disease despite two-drug therapy, particularly in bacteremic patients. Can J Microbiol, 1985 Jul, 31(7), 590 - 7 Chemical and enzymatic variation in the cell walls of pathogenic Candida species; Lyon FL et al.; Cell walls, isolated from seven pathogenic species of Candida, were lipid extracted and fractionated by treatment with ethylenediamine or enzymatically hydrolyzed using chitinase and laminarinase . Two different chitinase preparations were used, one from Streptomyces sp . which had some beta-1,3-glucanase activity, and another from Serratia marcescens which did not have glucanase activity . Laminarinase was a commercial preparation . The monosaccharide constituents of whole cell walls and the fractions derived from them were determined qualitatively and quantitatively by gas-liquid chromatography of the products of a mild acid hydrolysis and by the phenol - sulfuric acid assay of the products of a stronger acid hydrolysis . The monomeric constituents of the enzymatic hydrolyses were analyzed using gas-liquid chromatography . Approximately 50% of all walls was soluble in ethylenediamine . Glucose and mannose were the only monosaccharides found in all of the fractions derived from ethylenediamine extraction examined . Similarities among the strains, based upon relative amounts of glucose and mannose, were more apparent than differences, but statistical analyses of the data revealed a general trend of decreasing similarity in the following order, C . albicans and C . stellatoidea, C . tropicalis and C . parapsilosis, and C . pseudotropicalis, C . guilliermondii, and C . krusei . In the enzymatic assays, mannose and glucose were released by laminarinase, whereas glucose and N-acetyl-D-glucosamine or N-acetyl-D-glucosamine alone were released by the chitinases . These assays supported the trend in relationships cited above, with the data being somewhat more definitive. J Clin Microbiol, 1985 Jul, 22(1), 5 - 8 Isolation medium for the recovery of Pseudomonas cepacia from respiratory secretions of patients with cystic fibrosis; Gilligan PH et al.; A new medium for the isolation of Pseudomonas cepacia from respiratory tract secretions of patients with cystic fibrosis (CF) is described . This medium consists of inorganic salts, 0.5% pyruvate, and 0.1% proteose peptone as nutritive components and 0.0001% crystal violet, 0.15% bile salts, 100 micrograms of ticarcillin per ml, and 300 U of polymyxin B per ml as selective agents . The medium, designated PC medium, supported superior growth of 38 of 50 stock isolates of P . cepacia after 48 h of incubation when compared with MacConkey agar (0 of 50) . The medium completely inhibited the growth of 112 of 124 stock isolates of organisms commonly found in respiratory secretions of CF patients . Cultures were made on PC medium with respiratory secretions of 169 CF patients . P . cepacia was recovered from 35 patients with isolates on PC medium but from only 21 patients with isolates on MacConkey agar . Of 221 other potentially pathogenic isolates found in these specimens, only six (two Pseudomonas aeruginosa isolates, two molds, one yeast, and one Serratia marcescens isolate) grew on PC medium . PC medium should facilitate the recovery of P . cepacia from CF patients. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1985 Jul, 259(4), 461 - 7 Degradation of human fibronectin by metalloproteases of Serratia marcescens; Traub WH et al.; The purified metalloproteases of Serratia marcescens strains SF 178 and SH 186 attacked soluble human serum fibronectin . This observation points to a novel mechanism through which bacterial 'aggressins' might abolish nonspecific host defense mechanisms. Proc Natl Acad Sci U S A, 1985 Jul, 82(14), 4663 - 7 Translation activates the paused transcription complex and restores transcription of the trp operon leader region; Landick R et al.; It has been proposed that RNA polymerase pausing in the leader region of the tryptophan (trp) operon of Escherichia coli is responsible for the synchronization of transcription and translation essential to attenuation control . In this report we use an in vitro coupled transcription/translation system to study the effect of trp leader peptide synthesis on RNA polymerase pausing in the trp leader region . Wild-type and translation-defective trp leader templates of E . coli and Serratia marcescens were employed, and pause RNA synthesis and paused complex release (activation) were quantified relative to synthesis of the terminated leader transcript . It was observed that pausing in the trp leader region was prolonged when translation of the leader transcript was reduced by mutations in the leader region or by addition of the translation inhibitor kasugamycin or chloramphenicol . Experiments with S-30 extracts from a mutant strain that is inefficient in translating the tryptophan codons in the leader transcript indicated that ribosome movement to these codons also releases the paused transcription complex . These findings indicate that the paused trp leader transcription complex resumes transcription when released by ribosome movement over the leader peptide coding region . This release would facilitate the coupling of transcription and translation essential to attenuation control. Antimicrob Agents Chemother, 1985 Jul, 28(1), 107 - 12 Binding and neutralization of bacterial lipopolysaccharide by colistin nonapeptide; Warren HS et al.; Polymyxin nonapeptides, proteolytic derivatives of polymyxin antibiotics, are less toxic than their parent compounds but retain some of their antibacterial activities . To confirm and expand observations that polymyxin nonapeptides have anti-endotoxin activity, we studied the ability of colistin nonapeptide to bind to bacterial lipopolysaccharide (LPS) and to inhibit the effects of LPS on Limulus amoebocyte lysate and lymphocyte mitogenicity . Colistin nonapeptide was purified by high-pressure liquid chromatography and was demonstrated to bind to LPS by equilibrium dialysis . The ability of colistin nonapeptide to render E . coli ATCC 25922 cells sensitive to erythromycin was abrogated by 50% after incubation with E . coli O18 LPS in a ratio by weight of LPS to colistin nonapeptide of 3.9:1 . The presence of 4 micrograms of colistin nonapeptide or colistin per ml increased by 130- and 800-fold, respectively, the concentration of E . coli O113 LPS required to produce 50% gelation of Limulus amoebocyte lysate as measured by a spectrophotometric assay . Neutralization of LPS by colistin nonapeptide was time and concentration dependent . In contrast to the neutralization seen with LPS derived from a colistin-sensitive organism, colistin nonapeptide neutralized very little LPS extracted from a strain of Serratia marcescens that was resistant to colistin . Colistin nonapeptide also inhibited LPS-induced {3H}thymidine uptake by splenic lymphocytes, but its activity was less than 1/10 that of colistin . We conclude that colistin nonapeptide binds to LPS and possesses antiendotoxin activity . However, the anti-endotoxin activity of the nonapeptide is considerably less than that of its parent compound, colistin. Am J Med, 1985 Jun 7, 78(6A), 62 - 72 Potential of imipenem as single-agent empiric antibiotic therapy of febrile neutropenic patients with cancer; Wade JC et al.; Infection remains a major cause of morbidity and mortality for the patient with cancer who experiences episodes of severe granulocytopenia . The search continues for new antimicrobial agents with improved efficacy and lower incidence of toxicity . Imipenem is a new carbapenem antibiotic which possesses a broad antibacterial spectrum with excellent activity against Pseudomonas aeruginosa and the other commonly recovered enteric gram-negative bacilli that infect the granulocytopenic patient with cancer . The combination of imipenem plus an aminoglycoside has shown in vitro synergy against P . aeruginosa and Staphylococcus aureus whereas the combination of imipenem plus piperacillin or the extended spectrum cephalosporins have frequently shown antagonism when tested against P . aeruginosa and Serratia marcescens . The use of a P . aeruginosa-infected neutropenic rat model has provided an in vivo system to evaluate the activity of new antibiotics or antibiotic combinations . Monotherapy with imipenem is as effective in this model as any of the currently available synergistic antibiotic combinations . This degree of activity has not been found with other broad-spectrum antibiotics when used alone . Imipenem provides serum bactericidal activity well above a 1:8 dilution for the four most commonly isolated pathogens: P . aeruginosa, Escherichia coli, Klebsiella species, and S . aureus . In addition, imipenem's post-antibiotic effect against P . aeruginosa may be pertinent . Imipenem is a unique antibiotic, with properties that make it well suited for study as monotherapy for fever and suspected infection in granulocytopenic patients with cancer . A prospective randomized, double-blind study comparing imipenem with a control regimen of piperacillin plus amikacin as empiric antibiotic therapy of febrile granulocytopenic patients with cancer is currently underway at the University of Maryland Cancer Center. J Thorac Cardiovasc Surg, 1985 Jun, 89(6), 888 - 99 Deleterious effects of cardiopulmonary bypass . A prospective study of bubble versus membrane oxygenation; van Oeveren W et al.; A number of hematologic and immunologic parameters that reflect erythrocyte and platelet damage and host defense mechanisms against infection were studied in 20 patients undergoing cardiopulmonary bypass during coronary operations . The patients were randomly assigned to a group in which a bubble oxygenator or a hollow-fiber membrane oxygenator was used . Hemolysis, thrombocytopenia, and significant release of beta thromboglobulin occurred in patients from the bubble oxygenator group and, to much lesser extent, in patients from the membrane oxygenator group . Polymorphonuclear leukocytes and monocytes from bubble oxygenator patients demonstrated increased generation of reactive oxygen species in the resting state and in the presence of the stimulating agents N-formyl-methionyl-leucyl-phenylalanine, concanavalin A, and opsonized zymosan, as compared with cells from membrane oxygenator patients . No difference was found between bubble and membrane oxygenator patients in the time of occurrence or intensity of leukopenia during bypass, of leukocytosis at the end of bypass, nor in the rate of complement activation, as assessed by quantitation of plasma C3a antigen . Complement activation was dependent on the alternative pathway . Immunoglobulin M concentration significantly decreased during bypass in both groups of patients . The serum opsonizing capacity for endotoxin and serum bactericidal activity for Serratia marcescens were decreased in both groups, mainly because of hemodilution, although they were additionally affected by bubble oxygenation . Several deleterious hematologic consequences of cardiopulmonary bypass can be minimized by the use of a membrane oxygenator . However, complement activation remains a potential risk factor even in membrane oxygenator patients and requires further investigation to obtain better hemocompatible materials for cardiopulmonary bypass circuits. J Hosp Infect, 1985 Jun, 6(2), 218 - 20 An 'outbreak' of pulmonary pseudoinfection by Serratia marcescens; Siegman-Igra Y et al.; The recovery of multiple isolates of Serratia marcescens from bronchial lavage specimens was traced to contaminated fibreoptic bronchoscopes . Four patients were involved and none became infected . Awareness of a cluster of serratia cultures and immediate investigation and institution of control measures may have prevented the occurrence of true infections. Infect Immun, 1985 Jun, 48(3), 747 - 53 A serratial protease causes vascular permeability reaction by activation of the Hageman factor-dependent pathway in guinea pigs; Kamata R et al.; The 56-kilodalton (56K) protease isolated from a culture filtrate of Serratia marcescens caused vascular permeability enhancement followed by edema formation when injected into guinea pig peripheral corneas and subconjunctival space or skin . The character and the mechanism of permeability enhancement were analyzed in vivo . The enhancement was maximum at 5 to 10 min . The permeability reaction increased exponentially by the amount of enzyme used . The enhancement of permeability induced by the 56K protease was not affected by treatment with an antihistamine but was greatly augmented by simultaneous injection of a kinin potentiator, Glu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro-OH (SQ 20,881) . Furthermore, the permeability activity of the protease, but not the amidolytic activity, was inhibited by soybean trypsin inhibitor, a well-known inhibitor of plasma kallikrein, as well as by corn trypsin inhibitor, the best inhibitor of activated Hageman factor . Results of these in vivo studies indicate that the permeability-enhancing reaction induced by the 56K protease is caused by activation of the Hageman factor-dependent pathway in the tissue . The permeability-increasing activity of the 56K protease was parallel with the enzyme activity . Serratial lipopolysaccharide did not produce a permeability enhancement reaction within 30 min when injected into guinea pig skin . These results are consistent with the results of recent in vitro experiments in which activation of the purified Hageman factor but not of prekallikrein by the 56K protease was elucidated (Matsumoto et al., J . Biochem . (Tokyo) 96:739-749, 1984) . Thus, the molecular mechanism described above appears to be operative in the pathogenesis of corneal edema and chemosis, which is induced by S . marcescens, in addition to the direct tissue destruction by the protease. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1985 May, 259(3), 410 - 25 Active immunization of NMRI mice against Serratia marcescens; Traub WH et al.; Phenol-hot water lipopolysaccharide (LPS) extracts of Serratia marcescens strains CDC O3:H1, CDC O6:H3, NEW CDC O14:H12, and SH 186 (serotype O6/O14:H12) significantly protected NMRI mice against intraperitoneal challenge with the more mouse virulent homologous strains; overall, there was moderate cross-protection against the minority of heterologous challenge strains . Trichloroacetic acid LPS extracts and K-antigen extracts of strains NEW CDC O14:H12 and SH 186 also proved protective antigens . The purified metalloproteases of strains SH 186 and SF 178 (serotype O6/O14:H12) effected active murine immunization . Neither active nor passive immunization of NMRI mice with E . coli Rc mutant J5 afforded significant protection against various challenge strains of S . marcescens. Infection, 1985 May-Jun, 13(3), 140 - 5 Antiserum against Escherichia coli J5: a re-evaluation of its in vitro and in vivo activity against heterologous gram-negative bacteria; Trautmann M et al.; Antiserum against Escherichia coli J5, a "rough" mutant of E . coli 0111, has been reported to confer broad-spectrum protection against serologically unrelated gram-negative bacteria . In order to re-evaluate these findings, we examined the influence of rabbit antiserum against E . coli J5 on the phagocytosis of heterologous gram-negative bacteria by rabbit granulocytes in vitro and its ability to protect mice against gram-negative bacterial infection . In vitro, J5 antiserum enhanced the phagocytosis of E . coli 0111, E . coli 06 and Serratia marcescens 06/014:H2 when compared to normal rabbit serum . However, J5 antiserum did not enhance the phagocytosis of Klebsiella pneumoniae type 2 and Pseudomonas aeruginosa serotype 9 . In vivo, the protective effect of J5 antiserum against lethal gram-negative infection was not superior to that of normal (pre-immune) serum with the exception of E . coli 0111 septicemia . In contrast, type-specific antiserum against each of the smooth gram-negative bacteria markedly enhanced phagocytosis in vitro and exerted significant protection in vivo . Thus, in this study antiserum against E . coli J5 proved to be of limited value for opsonization of gram-negative bacteria and protection against gram-negative bacterial infection. Arch Microbiol, 1985 May, 141(4), 371 - 6 Hemolytic activity of Serratia marcescens; Braun V et al.; A cell-bound hemolytic activity was found in several strains of Serratia marcescens . One Serratia cell per ten erythrocytes was sufficient to cause complete lysis of human erythrocytes within 2 h in the liquid assay . The hemolytic activity resided in the membrane fraction and could be inactivated by incubating cells with proteases . The hemolytic activity was greatly enhanced in actively metabolizing Serratia cells and was partially controlled by the iron supply . Hemolysis was accompanied by degradation of erythrocyte membrane proteins (band 3 and 6, glycophorin) and was independent of the blood group . The exoprotease secreted by S . marcescens in large amounts was not involved in hemolysis . Comparison with various hemolytic strains of Escherichia coli showed that hemolysis of erythrocytes was more pronounced with S . marcescens than with E . coli . In contrast to hemolysis by E . coli, lysis of erythrocytes by S . marcescens was not enhanced by Ca2+ ions. J Antimicrob Chemother, 1985 May, 15(5), 559 - 65 Decreased susceptibility of Serratia marcescens to chlorhexidine related to the inner membrane; Lannigan R et al.; An isolate of Serratia marcescens (strain 100) obtained from a handwashing solution of chlorhexidine (Hibitane) was found not to release potassium ions when exposed to chlorhexidine as compared to chlorhexidine susceptible clinical isolates of Escherichia coli and Ser . marcescens . Lysozyme-tris-EDTA spheroplast preparations o