Staphylococcus aureus

Pediatr Res, 1986 Sep, 20(9), 843 - 7
Biosynthesis of variant medium chain acyl-CoA dehydrogenase in cultured fibroblasts from patients with medium chain acyl-CoA dehydrogenase deficiency; Ikeda Y et al.; We prepared monospecific antiserum in rabbits against medium chain acyl-CoA dehydrogenase (MCAD) purified from rat liver and studied the biosynthesis of MCAD in cultured skin fibroblasts from patients with MCAD deficiency using the antibody . Cells were incubated with {35S}methionine . The labeled MCAD was immunoprecipitated using the anti-rat MCAD antiserum and Staphylococcus aureus cells and then analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . We first demonstrated that antirat MCAD antibody crossreacted specifically with human MCAD . In 13 MCAD-deficient cell lines tested, the residual MCAD activity ranged from 5-12% of the mean of normal controls, but the variant MCAD in all of these cells was indistinguishable from the normal human MCAD on the basis of molecular size, indicating that MCAD deficiency in all of these patients is most likely due to point mutation(s) in the MCAD gene.

J Bacteriol, 1986 Sep, 167(3), 1016 - 9
Determination of hydrophobicity on bacterial surfaces by nonionic surfactants; Noda Y et al.; The hydrophobicity of the bacterial cell surface was determined by using nonionic surfactants . The method is based on the adsorption of nonionic surfactants at the hydrophobic sites of the cell surface . Among many nonionic surfactants, C18H37O(CH2CH2O)13H was preferred . The surfactant was added in excess to a bacterial suspension, and the suspension was mixed by sonication or mechanical stirring . The amount of surfactant remaining in the supernatant after centrifugation was determined spectrophotometrically by measuring the absorbance of tetrabromophenolphthalein ethylester . Effective dispersion of bacterial cells such as Staphylococcus aureus and Mycobacterium smegmatis was achieved by sonication in the presence of the nonionic surfactant . Adsorption measurements coincided with Langmuir's equation, indicative of monolayer adsorption . The method is useful for the determination of the hydrophobicity of various bacterial cell surfaces.

Antimicrob Agents Chemother, 1986 Sep, 30(3), 382 - 4
Efficacy of ciprofloxacin for experimental endocarditis caused by methicillin-susceptible or -resistant strains of Staphylococcus aureus; Carpenter TC et al.; The efficacy of ciprofloxacin for experimental aortic valve endocarditis in rabbits infected by either a methicillin-susceptible or a methicillin-resistant strain of Staphylococcus aureus was compared with standard therapy of nafcillin or vancomycin, respectively . After 4 days of therapy, ciprofloxacin reduced the counts of organisms in aortic valve vegetations as effectively as the standard regimen for both susceptible and resistant strains . Mean concentrations of ciprofloxacin in serum achieved 1 h after a dose exceeded the MBC for each strain by twofold or less . In these experiments ciprofloxacin was as efficacious as standard regimens currently used to treat staphylococcal infections in humans.

J Clin Microbiol, 1986 Sep, 24(3), 349 - 52
Detection of methicillin-resistant Staphylococcus epidermidis; Woods GL et al.; To determine whether methods suggested for detecting methicillin-resistant Staphylococcus aureus apply equally to methicillin-resistant Staphylococcus epidermidis, 135 S . epidermidis isolates were tested by the Vitek AMS gram-positive susceptibility card (Vitek Systems, Inc., Hazelwood, Mo.) and by modifications of agar screen, disk diffusion, and microdilution methods . Modifications included 24- versus 48-h incubation, unsupplemented versus 2% NaCl-supplemented broth, and standard versus direct inoculum . At 24 h, the highest number of resistant strains, 59, was detected by oxacillin (1 microgram) disk diffusion . At 48 h, three additional strains were judged resistant . With one exception, results for oxacillin disk diffusion and agar screen were equivalent at 24 and 48 h . Vitek detected 50 resistant strains . Significantly fewer resistant strains were detected at 24 h by methicillin disk diffusion (5 micrograms) and methicillin microdilution with 2% NaCl . For oxacillin microdilution, neither 2% NaCl supplementation nor the method of inoculum preparation significantly affected the results . Oxacillin microdilution with cation- rather than non-cation-supplemented broth detected significantly fewer (n = 33) resistant strains at 24 h; 51 were resistant at 48 h . To detect methicillin-resistant S . epidermidis, a direct inoculum with either 24-h oxacillin disk diffusion and reincubation of intermediate strains for an additional 24 h or 24-h oxacillin agar screen and reincubation of strains with no growth for a total of 48 h is recommended.

J Bacteriol, 1986 Sep, 167(3), 975 - 80
Molecular cloning of the gene of a penicillin-binding protein supposed to cause high resistance to beta-lactam antibiotics in Staphylococcus aureus; Matsuhashi M et al.; A novel penicillin-binding protein, PBP-2' (Mr about 75,000), is known to be induced in excessively large amount by most beta-lactam compounds in cells of a clinically isolated strain of Staphylococcus aureus, TK784, that is highly resistant to beta-lactams and also most other antibiotics . This protein has very low affinities to most beta-lactam compounds and has been supposed to be the cause of the resistance of the cells to beta-lactams . A 14-kilobase DNA fragment was isolated from the cells that carried the gene encoding this penicillin-binding protein and also a genetically linked marker that is responsible for the resistance to tobramycin . This DNA was cloned on plasmid pACYC184 and was shown to cause both production of PBP-2' and resistance to tobramycin in Escherichia coli cells . However, the formation of PBP-2' in E . coli was only moderate and was independent of normal inducer beta-lactams . The PBP-2' formed in the E . coli cells showed slow kinetics of binding to beta-lactams similar to that of PBP-2' formed in the original S . aureus cells and gave a similar pattern of peptides to the latter when digested with the proteolytic V8 enzyme of S . aureus.

Biochem J, 1986 Sep 1, 238(2), 561 - 70
A novel isoform of cytoplasmic actin that binds poly-L-proline; Kedersha NL et al.; An actin-like protein was purified to apparent homogeneity from chick-embryo homogenates and chick-embryo fibroblasts by the use of poly-L-proline-agarose affinity chromatography; we therefore refer to this protein as PBP (poly-L-proline-binding protein) . PBP binds to deoxyribonuclease-agarose, co-migrates with known actin standards on SDS/polyacrylamide-gel electrophoresis, and has an amino acid composition similar to that of actin . Linear peptide maps after digestion with Staphylococcus aureus proteinase reveal its apparent homology with gamma-actin; however, isoelectric-focusing experiments show that PBP is clearly more acidic than any of the three major isoforms of actin . PBP polymerizes in the presence of ATP to form fibrillar structures resembling actin paracrystalline aggregates . In chick-embryo fibroblasts, immunofluorescence with antibodies to PBP shows that its distribution is cytoplasmic: perinuclear staining of the cytoplasm, generalized cytoplasmic staining and peripheral fibrillar structures are evident . In contrast, antibodies specific for the (alpha, gamma)-actins reveal the typical stress fibre structures characteristic of fibroblastic cells . PBP appears to constitute a novel isoform of cellular actin, distinct from the known actin isoforms in terms of its lower isoelectric point, its ability to bind poly-L-proline and its distinct subcellular localization.

Arzneimittelforschung, 1986 Sep, 36(9), 1301 - 2
Bactericidal effect of combinations of cephalosporins with tobramycin on clinical isolates of Escherichia coli, Klebsiella and Staphylococcus aureus; Wagenvoort JH et al.; The antimicrobial activities of the cephalosporins cefazolin, cefotaxime, moxalactam or ceftazidime in combination with tobramycin against clinical isolates of E . coli, of Klebsiella, of beta-lactamase-negative and beta-lactamase-producing S . aureus strains were compared in vitro by the checkerboard technique . Antibiotic dilutions differed by small arithmetic increments . Favourable interactions occurred with each bacterial species . This trend applied to third-generation cephalosporins against E . coli and Klebsiella as well as to cefazolin against S . aureus . The antibiotics with already the highest intrinsic activity generally exhibited also the most favourable interaction.

Am J Vet Res, 1986 Sep, 47(9), 2017 - 9
Comparison of four test procedures to identify Staphylococcus aureus isolated from bovine intramammary infections; Hogan JS et al.; A comparison was made between conventional tube coagulase, macrocupsular coagulase, latex agglutination, and miniaturized biochemical test systems for identification of Staphylococcus aureus of bovine origin . A total of 303 gram-positive, catalase-positive cocci of bovine origin were tested . Agreement between each pair of 4-hour tube coagulase, macrocupsular coagulase, latex agglutination, and miniaturized biochemical test results within isolates was greater than 95.0 . Seventeen (5.6%) isolates were test negative for 4-hour tube coagulase, but test positive for 24-hour tube coagulase . Thirteen (76.5%) of these isolates were identified as S hyicus, 3 were S aureus, and 1 was not identified.

Nurs Res, 1986 Sep-Oct, 35(5), 263 - 8
Covergowns and the control of operating room contamination; Copp G et al.; This study assessed the effectiveness of cotton/polyester covergowns in protecting scrubsuits against bacterial contamination when operating room (OR) personnel are outside the clean environment of the operating suite . Rodac impression plates were used to measure bacterial contamination . The subjects were nurses working a normal daily OR routine . Bacterial colony counts on the right shoulder decreased when covergowns were worn over scrubsuits during the lunch period outside the OR and when fresh scrubsuits were put on following the lunch period . Colony counts rose over the lunch period when scrubsuits were worn unprotected outside the OR and when scrubsuits were removed before and put on again following lunch . Left thigh samples showed no significant effects of experimental treatments and yielded a mean colony count 2.8 times higher than right shoulder samples . Fifty-three percent of subjects were positive for Staphylococcus aureus and 16% yielded positive plates on 3 or more study days . The incidence of S . aureus contamination was affected by experimental treatments in a way similar to overall bacterial contamination . The results indicated that wearing covergowns protects against above-waist bacterial contamination of scrubsuits.

Infect Immun, 1986 Sep, 53(3), 663 - 70
Regulation of staphylococcal toxic shock syndrome toxin-1 and total exoprotein production by magnesium ion; Mills JT et al.; The effect of Mg2+ on in vitro production of extracellular proteins and, specifically, of toxic shock syndrome toxin-1 (TSST-1), by Staphylococcus aureus in a chemically defined medium was examined . As previously observed, the organisms did not proliferate in the absence of divalent cations . Low levels of Mg2+ (0.02 to 0.04 mM) permitted submaximal proliferation and elevated production of exoproteins . When the Mg2+ concentration was raised to 0.4 mM, multiplication was optimal and exoprotein levels were depressed . Ca2+ and Mn2+ diminished the effect of limiting Mg2+ . The increased levels of exoproteins were not due to cell lysis or leakage since intracellular TSST-1 levels were not high enough to account for the increase in extracellular TSST-1 and since the intracellular enzyme, lactate dehydrogenase, was not found in culture supernatants . Cells cultured in low levels of Mg2+ remained in logarithmic growth longer than did those cultured in high concentrations of Mg2+ and, unlike the latter, produced exoproteins throughout the logarithmic growth phase . Low Mg2+ had no effect on cultures in the stationary phase, and organisms cultured in low Mg2+ recovered fully when transferred to high Mg2+ . We conclude that, when cultured in medium deficient in Mg2+, S . aureus responds early in the growth cycle by increasing production of many extracellular proteins, including TSST-1.

Eur J Pediatr, 1986 Sep, 145(4), 252 - 7
In vitro analysis of lymphocyte functions in common variable immunodeficiency: heterogeneity in B-cell defects; Matsuoka H et al.; Ten patients with common variable immunodeficiency were classified into three groups according to the number of circulating B-cells, i.e . B-cells being absent (three patients), very low (three patients) or within the normal range (four patients) . The four patients in the last group showed significant proliferative responses to the T-independent B-cell mitogen, formalin-fixed Staphylococcus aureus, Cowan I . Further study of these patients by co-cultures with allogeneic T or B-cells in various combinations with pokeweed mitogen showed that two patients had an intrinsic B-cell defect without T-cell defect . The third patient had a T-cell dysfunction (i.e . his T-cells could only help the B-cells of some individuals) resulting in a defect in Ig production . The T-cells of the fourth patient showed poor helper function towards all controls . All six patients with absent or very low numbers of B-cells in group I and II had normal T-cell helper function . This study demonstrates that the immunological defect in common variable immunodeficiency is most often a B-cell defect at different stages of their differentiation with sometimes an additional T-cell dysfunction.

J Clin Microbiol, 1986 Sep, 24(3), 490 - 2
Failure of rapid agglutination methods to detect oxacillin-resistant Staphylococcus aureus; Ruane PJ et al.; Although a latex agglutination test (StaphAurex) and a hemagglutination test (Staphyloslide) correctly identified all strains of Staphylococcus aureus that were susceptible or had intermediate susceptibility to oxacillin, 17 of 73 (23%) and 18 of 73 (25%) strains of oxacillin-resistant S . aureus were not identified by StaphAurex and Staphyloslide, respectively . All strains not detected were resistant to trimethoprim-sulfamethoxazole and rifampin.

J Clin Invest, 1986 Sep, 78(3), 612 - 7
Staphylococcus aureus Cowan I . Potent stimulus of immunoglobulin M rheumatoid factor production; Levinson AI et al.; These studies demonstrate that Staphylococcus aureus Cowan I (SAC), a protein A-positive Staphylococcal strain, is a potent and consistent inducer of IgM rheumatoid factor production by normal human peripheral blood mononuclear cells . The frequency and magnitude of this response greatly exceeded that of parallel cultures stimulated with pokeweed mitogen or the protein A-negative S . aureus Wood strain, although all three agents induced a similar amount of total IgM . Cell fractionation studies indicated that SAC-induced IgM rheumatoid factor is T cell-dependent . The striking ability of SAC to induce IgM rheumatoid factor may relate to its protein A content, since cultures stimulated with protein A-coupled sepharose beads also consistently produced this autoantibody . Thus SAC is a new probe of in vitro IgM rheumatoid factor production and its use has provided further evidence that most healthy individuals harbor precursors of IgM rheumatoid factor secreting cells . Unlike other polyclonal activators, SAC is unique in its capacity to bind immunoglobulin, a property that may account for its prominent anti-IgG inducing capacity.

Blood, 1986 Sep, 68(3), 708 - 11
Granulocyte-macrophage colony-stimulating factor enhances phagocytosis of bacteria by human neutrophils; Fleischmann J et al.; In order to determine whether human granulocyte-macrophage colony-stimulating factor (GM-CSF) can enhance phagocytosis, neutrophils were combined with Staphylococcus aureus (S aureus), and both the number of bacteria per neutrophil and the percent of neutrophils phagocytizing were assessed in the absence and presence of GM-CSF . Exposure to GM-CSF did not enable neutrophils to ingest unopsonized bacteria . When bacteria were opsonized with serum, both the number of bacteria per neutrophil and the percent of cells phagocytizing were increased by treatment with GM-CSF . Digestion of extracellular organisms by lysostaphin was used to substantiate phagocytosis . These results indicate that another effect of GM-CSF on the mature neutrophil is the enhancement of phagocytosis.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Sep, (9), 29 - 32
{Determination of protein A I staphylococcus aureus strains isolated from monkeys}; Dzhikidze EK et al.; Protein A content in Staphylococcus aureus isolated from 6 species of monkeys at the Sukhumi Monkey Nursery has been studied . Protein A has been detected in 73% of the studied strains . One strain isolated from a rhesus macaque has been found to release high amounts of protein A into the environment.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1986 Sep, 262(3), 287 - 97
Biochemical properties of fibrinogen binding protein (clumping factor) of the staphylococcal cell surface; Usui Y; The staphylococcal fibrinogen binding protein of a strain of coagulase-negative Staphylococcus aureus was purified 229 fold in terms of the haemagglutination unit compared to the starting material by affinity chromatography on fibrinogen-Sepharose . By polyacrylamide gel electrophoresis in both reduced and unreduced gels, the protein showed one major band and minor bands with relative molecular masses of 62,000, 61,000, and 59,000, respectively . The isoelectric point was between 10.2 and 10.8 determined by isoelectric focusing in polyacrylamide gel . By the agar diffusion test one band was obtained against anti-whole cell rabbit serum . Amino acid analysis of fibrinogen binding protein showed glycine, glutamic acid, lysine, alanine, aspartic acid, and arginine as the major components . Protein A or teichoic acid extracted from the homologous strain did not show fibrinogen binding activity assayed by haemagglutination and anti-fibrinogen binding activity of anti-whole cell Fab . The amount of the fibrinogen binding protein to absorb the activity of anti-whole cell Fab was decreased to 1/60 of that of the starting material . Sheep red blood cells coated with the fibrinogen binding protein agglutinated in 0.001% (w/v) fibrinogen solution.

J Antibiot (Tokyo), 1986 Sep, 39(9), 1314 - 21
Studies on pristinamycin synergism in Staphylococcus aureus; Lacroix P et al.; Binding experiments were performed with both components of the pristinamycin complex (pristinamycin IA (PIA) and pristinamycin IIA (PIIA} using ribosomes from sensitive and resistant Staphylococcus aureus . Fluorescence polarization was used to measure PIA binding . The results obtained show a direct correlation between inhibition, synergy and the enhancement of the affinity of PIA for its receptor in the presence of PIIA . The uptake of PIA by intact cells seems to be directly correlated with affinity between PIA and ribosomes, a phenomenon which is probably shared with the macrolide antibiotics.

Res Vet Sci, 1986 Sep, 41(2), 191 - 5
Variations in immunoglobulin isotype produced during the antibody response to Brucella abortus and Staphylococcus aureus vaccines in sheep; Kerlin RL et al.; Experiments in sheep were carried out to examine factors modifying the immunoglobulin (Ig) isotype of the antibody response to Brucella abortus . Live B abortus (S19) stimulated higher titres of agglutinating antibody and IgG1 and IgG2 antibody than did killed B abortus . Live B abortus stimulated a more protracted synthesis of IgG2 antibody during the primary and secondary responses than did the killed S19 vaccine . In a second experiment, the capacity of live and killed Staphylococcus aureus to modify the antibody response to killed B abortus was examined . Both live and killed S aureus enhanced production of anti-brucella antibodies; this response was attributed to the adjuvant properties of S aureus . Killed S aureus enhanced production of anti-brucella antibody to a greater extent than live S aureus . Live S aureus did not preferentially enhance production of IgG2 anti-brucella antibody . The results suggested that the enhanced production of IgG2 antibody induced by live vaccines does not depend solely on a pyogenic lesion at the vaccination site.

J Antimicrob Chemother, 1986 Sep, 18(3), 359 - 64
Resistance of Mycoplasma pneumoniae to macrolides, lincomycin and streptogramin B; Stopler T et al.; Of four strains of Mycoplasma pneumoniae, highly resistant to erythromycin and related antibiotics, three were homogeneously resistant, but the fourth showed heterogeneous resistance, with only 1% of the cells manifesting this property . Stable, homogeneous resistance was generated in this strain in the presence of erythromycin, whereas the heterogeneous resistance was lost spontaneously on passage in the absence of antibiotics, or on treatment with acridine orange . The mechanism of induction of stable resistance appears to be different from that seen in Staphylococcus aureus.

J Med Microbiol, 1986 Sep, 22(2), 115 - 8
Drug resistance patterns and susceptibility to aflatoxin B1 of strains of Escherichia coli and Staphylococcus aureus; Tiwari RP et al.; The antibacterial properties of aflatoxin B1 have been evaluated against antibiotic-resistant clinical isolates of Escherichia coli and Staphylococcus aureus . The inhibition of growth ranged from 11.5 to 60.0% and 4.5 to 18.5% in the strains of S . aureus and E . coli, depending on the extent of drug resistance . Aflatoxin-B1 binding varied with toxin concentration, the presence of surfactants (Tween-80 or EDTA) as well as with the antibiotic-resistance pattern; binding was maximal in antibiotic-sensitive strains and least in the most resistant strains . Binding of aflatoxin B1, correlated with growth inhibition . Aflatoxin B1 also caused leakage of cell contents and decrease in inulin uptake, effects which were also concentration dependent.

J Bacteriol, 1986 Sep, 167(3), 888 - 92
Nucleotide sequence of the constitutive macrolide-lincosamide-streptogramin B resistance plasmid pNE131 from Staphylococcus epidermidis and homologies with Staphylococcus aureus plasmids pE194 and pSN2; Lampson BC et al.; The complete nucleotide sequence of the Staphylococcus epidermidis plasmid pNE131 is presented . The plasmid is 2,355 base pairs long and contains two major open reading frames . A comparison of the pNE131 DNA sequence with the published DNA sequences of five Staphylococcus aureus plasmids revealed strong regional homologies with two of them, pE194 and pSN2 . The region of pNE131 containing the reading frame which encodes the constitutive ermM gene is almost identical to the inducible ermC gene region of pE194, except for a 107-base-pair deletion which removes the mRNA leader sequence required for inducible expression . A second region of pNE131 contains an open reading frame with homology to the small cryptic plasmid pSN2 and potentially encodes a 162-amino-acid protein.

J Antimicrob Chemother, 1986 Sep, 18(3), 301 - 6
Uptake of gentamicin by Staphylococcus aureus possessing gentamicin-modifying enzymes: enhancement of uptake by puromycin and N,N'-dicyclohexylcarbodiimide; Gilman S et al.; Uptake of gentamicin by a gentamicin-resistant strain of Staphylococcus aureus possessing the aminoglycoside-modifying phosphotransferase enzyme APH(2") was enhanced by the protein synthesis inhibitor, puromycin or by the proton-translocating ATPase inhibitor, N,N'-dicylohexylcarbodiimide . Such enhanced uptake was inhibited by carbonyl cyanide p trifluoromethoxyphenylhydrazone or by valinomycin in the presence of potassium ions, suggesting a role for the transmembrane proton motive force in the process . The accumulated gentamicin did not cause loss of cell viability and exhibited altered chromatographic mobility compared with a control (unmodified) preparation of gentamicin.

J Biomed Mater Res, 1986 Sep, 20(7), 989 - 1002
The biological performance of calcium phosphate ceramics in an infected implantation site: I . Biological performance of hydroxyapatite during Staphylococcus aureus infection; van Blitterswijk CA et al.; In the present study the biological performance of macroporous and dense hydroxyapatite after implantation in the rat middle ear was evaluated during an induced Staphylococcus aureus middle ear infection . The course of the infection was similar to that in the absence of an implant . Hydroxyapatite was frequently integrated with fibrous ingrowths in the middle ear lumen, originating solely from the infection . Good epithelial covering of the implant with all types of epithelial cells of importance for middle ear defence, was found . Increase of the exudate in the pores due to the infection was relatively small, and most of the exudate was restricted to pores on the implant surface . The bony tissue in the pores was not influenced significantly by the induced infection . Degradation of hydroxyapatite was consistent with earlier results obtained in the non-infected middle ear . The results obtained so far suggest that hydroxyapatite is highly suitable for middle ear implantation.

J Biomed Mater Res, 1986 Sep, 20(7), 1003 - 15
The biological performance of calcium phosphate ceramics in an infected implantation site: II . Biological evaluation of hydroxyapatite during short-term infection; van Blitterswijk CA et al.; Macroporous hydroxyapatite was implanted submucosally in the rat middle ear and studied after intratympanic injection of a Staphylococcus aureus suspension . The middle ear infection was induced 1 week after the implantation, and the effects of infection on the middle ear and the implant material were evaluated after 1, 3, 7, and 14 days by light and electron microscopy . The findings in the infected middle ear with an implant corresponded well with those described for the infected middle ear cavity without an implant . The reactions of the tissue over the implant were similar to those of the original mucosa of the middle ear . Bone was deposited on the implant and in its pores in relatively large quantities . Biodegradation, due at least partially to phagocytic activity of macrophages and multinucleated cells, was more prominent than previously found . This higher degree of biodegradation may be attributed to the use of the mucosal implantation technique, because this was the only point of divergence with respect to material or methods from earlier work reported by our group . The present results, together with those published earlier, suggest that this material has promising features for use as a bone substitute in reconstructive middle ear surgery . Definitive conclusions on biological performance and biofunctionality will, however, have to await long term clinical trials.

Pediatr Res, 1986 Sep, 20(9), 848 - 52
Effect of the synthetic inhibitor tosylamino-phenylethyl-chloromethylketone on chemotactic peptide receptor activation and superoxide production in human neutrophils; Suter S et al.; It was previously shown that inhibitors such as tosylamido-phenylethyl-chloromethylketone (TPCK) inhibit superoxide production by human neutrophils . These studies suggested that a chymotrypsin-like protease inhibited by TPCK was involved in the activation of the neutrophils oxidative system . In this study, we attempted to define the step in cellular activation and/or cell function inhibited by TPCK . TPCK 10(-5) M did not inhibit the following early events thought to be involved in the activation of oxidase . 1) f-met-leu-phe-induced activation of phospholipase C assessed by the production of inositol-tris-phosphate (IP3), 2) f-met-leu-phe-induced membrane potential changes, 3) f-met-leu-phe-induced increase in free cytosolic calcium, and 4) phorbol-myristate acetate-induced protein phosphorylation in 32P labeled neutrophils . We also showed that TPCK 10(-5) M inhibited bactericidal activity of neutrophils on Staphylococcus aureus, whereas it did not inhibit the ingestion rate of endotoxin-coated Oil red O particles . We conclude that 1) TPCK at the concentration of 10(-5) M inhibits superoxide production but not ingestion of Oil red O particles and 2) TPCK inhibits superoxide production at a step distal from calcium mobilization and protein phosphorylation . Radiolabeled TPCK may therefore be a useful tool to study, whether a protease is involved in the activation of the oxidative system distal to second messenger generation.

Proc Natl Acad Sci U S A, 1986 Sep, 83(18), 6980 - 4
The murine Fc receptor for immunoglobulin: purification, partial amino acid sequence, and isolation of cDNA clones; Hibbs ML et al.; The murine Fc receptor for IgG (Fc gamma R) was purified to homogeneity by immunoaffinity chromatography from detergent lysates of the macrophage cell line J774 . Microsequencing of intact protein yielded a single amino-terminal sequence, which was confirmed and extended to 20 residues by the isolation of an overlapping peptide . The isolation of additional proteolytic fragments obtained by using Staphylococcus aureus V8 protease, cyanogen bromide, and lysine C proteinase, facilitated sequence analysis of a total of 119 amino acid residues . Codon usage charts were used to construct oligonucleotide probes based on the amino acid sequences of three nonoverlapping peptides . These probes were used to screen a cDNA library derived from the WEHI-3B myelomonocytic cell line, and a single cDNA clone (pFc24) to which all three probes hybridized was isolated . This clone, containing a 1.02-kilobase cDNA insert, has been characterized by restriction mapping and partial DNA sequencing, and it has been shown to encode the Fc gamma R . The sequence at the 5' end of the clone contained the coding information for the amino-terminal sequence of the Fc gamma R as well as a putative 13-amino acid signal sequence . The 3' end of the clone encoded a peptide identified in purified receptor preparations . Thus, the presence of coding information at the 5' and 3' ends of this clone suggests that full-length Fc receptor cDNA spans greater than 1 kilobase.

J Hosp Infect, 1986 Sep, 8(2), 143 - 8
Methicillin-resistant Staphylococcus aureus in the UK and Ireland . A questionnaire survey; Cooke EM et al.; The results of a questionnaire survey of the distribution of methicillin-resistant Staphylococcus aureus (MRSA) in the UK and Ireland between 1982 and 1983 are reported . Information was obtained about the geographical distribution of MRSA, the units affected, the sites of isolation and the preventive measures employed . Serious clinical problems were confined to a small number of hospitals with high isolation rates of MRSA.

J Biochem (Tokyo), 1986 Sep, 100(3), 651 - 61
Antigenic structure of adenylate kinase from porcine skeletal muscle . IV . Two antigenic determinants on carboxyl-terminal peptide 126-194; Endo S et al.; Delineation of the location(s) of antigenic activity in CNBr peptide 126-194 from porcine skeletal muscle adenylate kinase (AK) was attempted . Peptide 126-194 was digested with chymotrypsin, Staphylococcus aureus V8 protease and trypsin, and several short peptides were purified from the digests by reverse-phase high-performance liquid chromatography (HPLC) . Inhibition of the binding of radioiodinated peptide 126-194 to goat antibody to porcine skeletal muscle AK (anti-AK antibody) by the peptides obtained by the enzymatic cleavages was examined by solid phase radioimmunoassay (RIA) . At least two antigenic determinants have been identified from the results . One is in the amino (N)-terminal half region 126-154, especially in the vicinity of 131-144, and the other is in the carboxyl (C)-terminal half region 165-183, especially in the vicinity of 165-171 . Both of them seem to correspond to exposed and accessible regions in the three-dimensional structure of AK . The correlation between antigenicity and high mobility of the loop in the estimated antigenic region 131-144 is also discussed.

J Biol Chem, 1986 Aug 25, 261(24), 11334 - 40
Comparisons of antibody reactivity and enzyme sensitivity between small proteoglycans from bovine tendon, bone, and cartilage; Vogel KG et al.; Preparations of small proteoglycans from bovine tendon, bone, and cartilage have been compared for sensitivity to various enzymes and reactivity with different polyclonal antibodies . Chondroitinase ABC digestion of all proteoglycans generated a core protein preparation that migrated similarly in sodium dodecyl sulfate-polyacrylamide electrophoresis as a doublet band with Mr approximately equal to 45,000 . The small proteoglycans of cartilage were divided into two populations based upon electrophoretic migration of the intact molecules (Rosenberg, L . C., Choi, H . U., Tank, L-H., Johnson, T . L., Pal, S., Webber, C., Reiner, A., and Poole, A . R . (1985) J . Biol . Chem . 260, 6304-6313) . The core preparations of tendon, bone, and the faster-migrating (PG II) proteoglycans of cartilage all interacted in Western blot/enzyme-linked immunosorbent assay analysis with polyclonal antibody raised against either the tendon or bone proteoglycans . The slower-migrating (PG I) proteoglycans of cartilage did not react with these antibodies . Digestion of the tendon small proteoglycan with Staphylococcus aureus V8 protease released glycosaminoglycan chains from the molecule and generated a 40-kDa protein fragment that was resistant to further rapid degradation by the enzyme . This large digestion fragment was also prominent following V8 protease digestion of the faster-migrating (PG II) population of small cartilage proteoglycans, but not the small proteoglycan of bone . The N-terminal amino acid sequence of the tendon (PG II) proteoglycan was determined . These observations provide additional evidence for heterogeneity among the chemically similar small proteoglycans from different tissues.

J Biol Chem, 1986 Aug 25, 261(24), 11315 - 9
Sensitization of the Escherichia coli cyclic AMP receptor protein to trypsin cleavage by polydeoxyribonucleotides and polyribonucleotides; Angulo J et al.; In the absence of cAMP the cyclic AMP receptor protein (CRP) is relatively resistant to trypsin whereas the cAMP X CRP complex is attacked yielding N-terminal core fragments of 14,300 and 18,500 Da which still bind cAMP . The DNA X CRP complex formed at low ionic strength in the absence of cAMP is cleaved by trypsin with the formation of 9,700- and 6,000-Da fragments and the concomitant loss of cAMP binding activity . DNA X CRP remains as resistant to attack by subtilisin, clostripain, and the Staphylococcus aureus V8 protease as unliganded CRP but is slowly digested by chymotrypsin . All of the double-stranded polydeoxyribonucleotides and several of the single-stranded polydeoxyribonucleotides and polyribonucleotides tested render CRP sensitive to cleavage by trypsin . CRP is less rapidly cleaved by trypsin in the presence of d(A)n, d(I)n, and r(C)n indicative of a weaker affinity of CRP for these polynucleotides . The 9,700-Da fragment is N-terminal in CRP and probably terminates at Lys-89 . The loss of cAMP binding activity following trypsin cleavage of DNA X CRP indicates that regions beyond this residue are important in the function of the cAMP-binding domain of CRP . The 6,000-Da fragment extends from Val-131 to Arg-185 or Lys-188 and contains part of the F helix involved in DNA binding by CRP.

J Biol Chem, 1986 Aug 25, 261(24), 11290 - 4
Functional domains of assimilatory NADH:nitrate reductase from Chlorella; Solomonson LP et al.; Assimilatory nitrate reductase from Chlorella is a homotetramer which contains one of each of the prosthetic groups FAD, heme, and molybdenum per subunit . Besides the reduction of nitrate by NADH, nitrate reductase also catalyzes the partial activities NADH:cytochrome c reductase, NADH:ferricyanide reductase, and reduced methyl viologen:nitrate reductase . Incubation of native nitrate reductase with either trypsin, Staphylococcus aureus V8 protease, or a natural inactivator protease from corn results in a loss of NADH:nitrate reductase and NADH:cytochrome c reductase activities but no loss of reduced methyl viologen:nitrate reductase activity . Incubation of nitrate reductase with V8 protease or corn inactivator protease resulted in two different products, each of which retained a different partial activity . Reduced methyl viologen:nitrate reductase activity was associated with a homotetrameric fragment of about 260 kDa which contained heme and molybdenum but no FAD . The molecular mass of native nitrate reductase determined under the same conditions was 375 kDa . NADH:ferricyanide reductase activity was associated with a monomeric species of approximately 30 kDa which contained FAD and the NADH-binding site . These results are consistent with a structure-function model of nitrate reductase which has the following features: FAD/NADH-binding domains exposed on the surface of the molecule, a protease-sensitive hinge region which connects the nitrate-reducing and NADH dehydrogenase moieties, and the quaternary structure maintained via association sites on the heme/molybdenum domain.

J Biol Chem, 1986 Aug 15, 261(23), 10945 - 51
Topology of mannosidase II in rat liver Golgi membranes and release of the catalytic domain by selective proteolysis; Moremen KW et al.; The orientation of mannosidase II, an integral Golgi membrane protein involved in asparagine-linked oligosaccharide processing, has been examined in rat liver Golgi membranes . Previous studies on mannosidase II purified from Golgi membranes revealed an intact subunit of 124,000 daltons, as well as a catalytically active 110,000-dalton degradation product generated during purification (Moremen, K . W., and Touster, O . (1985) J . Biol . Chem . 260, 6654-6662) . In Triton X-100 extracts of Golgi membranes, the intact enzyme was cleaved by a variety of proteases to generate degradation products similar to those observed previously . At appropriate concentrations, chymotrypsin, pronase, and proteinase K generated 110,000-dalton species, while trypsin and Staphylococcus aureus V8 protease generated 115,000-dalton forms . Cleavage by chymotrypsin under mild conditions (10 micrograms/ml, 10 min, 20 degrees C) resulted in a complete conversion to a catalytically active 110,000-dalton form of the enzyme which was extremely resistant to further degradation . Attempts to demonstrate these protease digestions in nonpermeabilized Golgi membranes were unsuccessful, a result suggesting that the protease-sensitive regions are not accessible on the external surface of the membrane . In Golgi membranes permeabilized by treatment with 0.5% saponin, mannosidase II could readily be cleaved to the 110,000-dalton form by digestion with chymotrypsin under conditions similar to those which result in a proteolytic inactivation of galactosyltransferase, a lumenal Golgi membrane marker . Although mannosidase II catalytic activity was not diminished by this chymotrypsin digestion, as much as 90% of the enzyme activity was converted to a nonsedimentable form . To examine the effect of the proteolytic cleavage on the partition behavior of the enzyme, control and chymotrypsin-treated Triton X-114 extracts of Golgi membranes were examined by phase separation at 35 degrees C . The undigested enzyme partitioned into the detergent phase consistent with its location as an integral Golgi membrane protein, while the 110,000-dalton chymotrypsin-digested enzyme partitioned almost exclusively into the aqueous phase in a manner characteristic of a soluble protein . These results suggest that mannosidase II catalytic activity resides in a proteolytically resistant, hydrophilic 110,000-dalton domain . Attachment of this catalytic domain to the lumenal face of Golgi membranes is achieved by a proteolytically sensitive linkage to a 14,000-dalton hydrophobic membrane anchoring domain.

J Biol Chem, 1986 Aug 15, 261(23), 10526 - 33
Structure of hemocyanin II from the horseshoe crab, Limulus polyphemus . Sequences of the overlapping peptides, ordering the CNBr fragments, and the complete amino acid sequence; Nakashima H et al.; The amino acid sequence of the largest fragment, CNBr Ia (203 residues) has been reported (Yokota, E., and Riggs, A . F . (1984) J . Biol . Chem . 259, 4739-4749) . The amino acid sequences of the second largest fragment, CNBr Ib (142 residues), and of the 12 smaller fragments are reported in accompanying papers (Moore, M . D., Behrens, P . Q., and Riggs, A . F . (1986) J . Biol . Chem . 261, 10511-10519; Behrens, P . Q., Nakashima, H., and Riggs, A . F . (1986) J . Biol . Chem . 261, 10520-10525) . The complete amino acid sequence of hemocyanin component II has been established by isolation and analysis of 13 methionine-containing peptides from either a tryptic digest or a Staphylococcus aureus strain V8 protease digest of whole carboxamidomethylated hemocyanin II . Hemocyanin II is composed of 628 residues and has a molecular weight with two copper atoms of 72,946.

Biochem J, 1986 Aug 15, 238(1), 65 - 73
Substrate specificity and characterization of rat liver p-nitrophenol, 3 alpha-hydroxysteroid and 17 beta-hydroxysteroid UDP-glucuronosyltransferases; Falany CN et al.; Purified preparations of rat liver 17-hydroxysteroid, 3-hydroxyandrogen and p-nitrophenol (3-methylcholanthrene-inducible) UDP-glucuronosyltransferases were further characterized as to their substrate specificities, phospholipid-dependency and physical properties . The two steroid UDP-glucuronosyltransferases were shown to exhibit strict stereospecificity with respect to the conjugation of steroids and bile acids . These enzymes have been renamed 17 beta-hydroxysteroid and 3 alpha-hydroxysteroid UDP-glucuronosyltransferase to reflect this specificity for important endogenous substrates . An endogenous substrate has not yet been identified for the p-nitrophenol (3-methylcholanthrene-inducible) UDP-glucuronosyltransferase . The steroid UDP-glucuronosyltransferase activities were dependent on phospholipid for maximal catalytic activity . Complete delipidation rendered the UDP-glucuronosyltransferases inactive, and enzymic activity was not restored when phospholipid was added to the reaction mixture . After partial delipidation, phosphatidylcholine was the most efficient phospholipid for restoration of enzymic activity . Partial delipidation also altered the kinetic parameters of the 3 alpha-hydroxysteroid UDP-glucuronosyltransferase . The three purified UDP-glucuronosyltransferases are separate and distinct proteins, with different amino acid compositions and peptide maps generated by limited proteolysis with Staphylococcus aureus V8 proteinase . Some similarity was observed between the amino acid composition and limited proteolytic maps of the steroid UDP-glucuronosyltransferases, suggesting they are more closely related to each other than to the p-nitrophenol UDP-glucuronosyltransferase.

J Immunol, 1986 Aug 15, 137(4), 1208 - 13
Identification of an early activation antigen (Bac-1) on human B cells; Suzuki T et al.; We have produced a monoclonal antibody, Bac-1, that appears to identify a novel antigen on activated human B cells . The Bac-1 antigen can be detected between 8 to 16 hr, as well as transferrin receptors (T9), after activation of small resting B cells with phorbol myristic acetate, anti-IgM antibody, Staphylococcus aureus Cowan I, or Epstein-Barr virus . The expression of the Bac-1 antigen precedes that of IL 2 receptors (Tac-1) . Peak expression of the Bac-1 antigen was observed on day 3 after activation, and decreased thereafter . The Bac-1 antigen was present on a minor subpopulation of relatively large B cells isolated from blood samples, and on "preactivated" B cells of heterogeneous size isolated from spleens and tonsils . It was not detected on bone marrow pre-B cells, blood small B cells, or plasma cells, nor was it expressed by resting or activated T cells or nonlymphoid cells . Certain B cell neoplasms and B lymphoblastoid cell lines were Bac-1+, but neoplastic cells of non-B lineage were Bac-1- . With immunoperoxidase staining, Bac-1+ cells were detected predominantly in the germinal centers of tonsil sections . The Bac-1 antigen on activated B cells was destroyed by protease treatment and was enhanced by neuraminidase treatment, suggesting that the Bac-1 antibody detects a cell surface molecule via an antigenic determinant which is partially obscured by neighboring sialic acid residues . The reactivity pattern of Bac-1 differs from the patterns of cellular reactivity reported for other monoclonal antibodies with specificity for activated human B cells.

Eur Heart J, 1986 Aug, 7(8), 679 - 84
Treatment of Staphylococcus aureus endocarditis: an analysis based on 25 proven cases; Freeman R et al.; Twenty-five consecutive unequivocal cases of Staphylococcus aureus endocarditis in a single centre over 5 years were reviewed . Despite an unusually high proportion of prosthetic infections an early mortality of only 16% was observed . Analysis reveals that early diagnosis and prompt effective antibiotic chemotherapy were important factors in achieving this low mortality . Monotherapy with an isoxazolyl penicillin (flucloxacillin) was shown to be as effective as combination therapies, and there was a significant association between the use of aminoglycosides and subsequent renal damage.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Aug, (8), 85 - 8
{Comparative evaluation of the diagnostic effectiveness of staphylococcal allergens of different manufacture in bronchospastic reactions of sensitized lungs}; Shamsutdinova GS et al.; With allergic immediate hypersensitivity test to the antigens of Staphylococcus aureus strain Wood 46 specific bronchospastic reactions were simulated in the lungs of guinea pigs by means of two allergens: commercial allergen manufactured in Czechoslovakia and staphyloallergen prepared by ultrasonic treatment under laboratory conditions at the Alma-Ata Research Institute of Epidemiology, Microbiology and Infectious diseases (USSR) . Allergens prepared at the Kazan Research Institute of Epidemiology and Microbiology (USSR) and in Bulgaria produced no reactions.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Aug, (8), 14 - 8
{Comparative immunochemical characteristics of extracellular polysaccharide-containing preparations and teichoic acids of Staphylococcus aureus}; Iastrebova NE et al.; The sensitivity of the enzyme immunoassay on the basis of polysaccharide-containing preparations was found to be lower than that of the assay on the basis of teichoic acids . The specificity of both assays was 100% . There was no complete coincidence between the results of the detection of antibodies to these two antigens in patients' sera.

Scand J Haematol, 1986 Aug, 37(2), 162 - 7
Hyperactive phagocytosis by circulating neutrophils and monocytes in Chédiak-Higashi syndrome; Komiyama A et al.; Phagocytic capacity of circulating neutrophils and monocytes was studied by electron microscopy in 2 children with Chediak-Higashi syndrome (CHS) . Apparent phagocytosis of autologous leucocytes by the CHS neutrophils and monocytes was demonstrated when they were coincubated with Staphylococcus aureus in vitro, indicating their capacity for hyperactive phagocytosis . The hyperactive phagocytic capacity of the CHS phagocytes was a cellular abnormality . CHS neutrophils and monocytes are known to be defective in chemotaxis and bactericidal capacity, and the hyperactive phagocytic capacity is another functional abnormality of CHS phagocytes.

Neth J Surg, 1986 Aug, 38(4), 118 - 20
Bacterially contaminated detached autogenous bone fragments--an experimental study; Stroosma OC et al.; Bacterially contaminated and uncontaminated standardized autogenous bone fragments were compared in an experimental canine model . Bacterial contamination was effected by immersing in a Staphylococcus aureus suspension of 6 X 10(8)/ml for 15 minutes . The fragments were replaced and either left detached or fixed with a lag screw . In a number of cases bone replacement was combined with transverse osteotomy followed by reduction and fixation with a six-hole neutralization plate based on the AO-principle . Follow-up carried out after six and 12 weeks, comprised radiological, histological, microradiographic, fluorescence microscopic and micro-angiographic investigation . Non-quantitative assessment indicated that the rate of resorption and new bone formation in the bacterially contaminated fragments definitely exceeded that of the uncontaminated fragments . Semi-quantitative findings seemed to confirm these conclusions . After 12 weeks this difference had virtually disappeared, except in loosely replaced infected fragments, which still showed more resorption and new bone formation . Integration of all fragments was good, there was no encapsulation with sequestration.

J Am Acad Dermatol, 1986 Aug, 15(2 Pt 1), 192 - 7
Density of the microflora in hand eczema before and after topical treatment with a potent corticosteroid; Nilsson E et al.; Twenty patients with hand eczema were studied with the use of quantitative bacteriologic cultures before and after successful topical treatment with a potent corticosteroid . One sample was taken from the most pronounced eczematous lesion, a second from skin affected with only erythema, and a third from clinically normal skin . Before quantitative bacteriologic analysis, the different species were combined into three main groups: Staphylococcus aureus, other aerobes, and anaerobes . Before treatment, S . aureus colonized the most pronounced eczematous lesion in eighteen of twenty patients and exceeded 10(5) cfu/cm2 in fifteen of twenty patients . The geometric mean count of S . aureus before treatment was significantly higher in eczema (10(4.8) cfu/cm2) than in erythema (10(3.4) cfu/cm2) and significantly higher in erythema than in normal skin (10(1.7) cfu/cm2) . The density of other aerobes and anaerobes was similar in the three sampling sites . After treatment, the mean count of S . aureus was significantly reduced at all three sampling sites, and the densities became equal . Treatment did not affect the mean count of other aerobes or anaerobes.

J Med Microbiol, 1986 Aug, 22(1), 9 - 15
Isolation and characterisation of a family of small plasmids encoding resistance to nucleic acid-binding compounds in Staphylococcus aureus; Emslie KR et al.; A family of small plasmids encoding resistance to nucleic acid-binding (NAB) compounds has recently been identified in strains of Staphylococcus aureus isolated in Italy, Texas and Western Australia . The mol . wts of the NAB-resistance plasmids are in the range (1.5-1.9) X 10(6) and all but one encode resistance to acridine yellow, ethidium bromide and quaternary ammonium compounds . The largest of the plasmids, pWG1773, differed in that it did not confer resistance to ethidium bromide . Restriction enzyme analysis of these plasmids revealed four distinct patterns corresponding to plasmids of four different mol . wts and physical maps were constructed based on the restriction patterns . Two plasmid types of molecular sizes approximately 2440 and 2240 base pairs had a 610-base pair region in common . Physical maps of the other two plasmid types were not related . The presence of a family of small NAB-resistance plasmids which carry no other known phenotypic markers provides further evidence for the strong selective advantage associated with maintenance of this determinant in clinical isolates of S . aureus.

J Bacteriol, 1986 Aug, 167(2), 726 - 8
Sequence of the exfoliative toxin B gene of Staphylococcus aureus; Jackson MP et al.; We sequenced the Staphylococcus aureus exfoliative toxin B gene contained on a 1.7-kilobase HindIII fragment of plasmid pRW001 . The gene was located by comparison of the amino acid sequences of open reading frames with the amino-terminal sequence of exfoliative toxin B and the total amino acid composition of the protein (A.D . Johnson, L . Spero, J.S . Cades, and B.T . De Cicco, Infect . Immun . 24:679-684, 1979) . The primary translation product consists of 274 amino acids and contains a 31-amino-acid N-terminal peptide presumably necessary for transport.

Infect Immun, 1986 Aug, 53(2), 441 - 4
Immunological protection of rabbits infected with Staphylococcus aureus isolates from patients with toxic shock syndrome; Scott DF et al.; Toxic shock syndrome toxin-1 (TSST-1) isolated from the growth medium of Staphylococcus aureus 1169 and 555 was used to immunize male rabbits before infection with either a TSST-1+ or a TSST-1- strain of S . aureus isolated from cases of TSS . None of the immunized rabbits died as a result of the infections, whereas 50% of the nonimmunized rabbits infected with the TSST-1- strain, D4508, and 75% of those infected with the TSST-1+ strain, 555, died . Western blots of crude extracellular protein preparations probed with sera from immunized rabbits indicated that the TSST-1- strain produces a 30,000-molecular-weight protein that cross-reacts with antiserum to TSST-1 . Because both organisms caused similar diseases in rabbits, we propose to designate the cross-reacting protein as TSST-2.

Chest, 1986 Aug, 90(2), 293 - 5
Endocarditis of a tricuspid prosthesis causing valvular stenosis and shunting through a patent foramen ovale; Bier A et al.; A young intravenous drug user presented with Staphylococcus aureus endocarditis involving the tricuspid valve, which was replaced with a Hancock bioprosthesis . She presented again with fever and dyspnea five months later and was found to be cyanotic . Recurrent endocarditis involving the prosthesis with right-to-left shunting through a patent foramen ovale was documented by echo and confirmed at autopsy.

Clin Pediatr (Phila), 1986 Aug, 25(8), 395 - 9
Neonatal mastitis; Walsh M et al.; Forty-one cases of neonatal mastitis seen at Children's Hospital, Boston since 1947 have been analyzed and the literature since 1950 reviewed . All 41, like those in the literature, occurred in full-term infants 1-5 weeks of age, with a sex ratio of 2:1 (females:males) . Bilaterality was rare (3) cases in this series, one in the literature review) and systemic spread or extramammary foci even rarer . The incidence has changed little in the past 35 years except for the larger number of cases in the 1950s . In the present series, all but a few cases have been caused by Staphylococcus aureus, and gram-negative enteric bacilli have not been seen . Therapy is surgical incision and drainage when fluctuance is present, but early treatment with appropriate intravenous antibiotics has apparently obviated the need for surgery in many recent cases . The prognosis for cure of the infection is excellent.

Clin Orthop, 1986 Aug, (209), 249 - 54
Prophylaxis with cefamandole nafate in elective orthopedic surgery; Henley MB et al.; A prospective, randomized, double-blind study was conducted to determine the efficacy of cefamandole nafate in reducing infections in general orthopedic procedures . Of 743 patients initially entered into the study, 715 (362 on cefamandole, 353 on placebo) fulfilled the requirements of the protocol . The infection rate was 1.6% for the cefamandole-treated group and 4.2% for the placebo group . In operations lasting longer than two hours, there were two infections in the cefamandole group and seven infections in the placebo group (p less than 0.05) . Staphylococcus aureus and gram-negative bacilli were the common pathogens . Adverse side effects were limited to transient elevations in liver enzymes.

J Infect Dis, 1986 Aug, 154(2), 273 - 82
Antigen-induced experimental septic arthritis in rabbits after intraarticular injection of Staphylococcus aureus; Mahowald ML et al.; With the Dumonde-Glynn model of antigen-induced arthritis, a rabbit model was developed to examine the histopathologic differences between normal and arthritic joints in the same animal infected by intraarticular injections of Staphylococcus aureus . Microscopic examination of whole joint sections and a quantitative histopathologic scale were used to compare changes in all the articular components of 17 normal and 17 arthritic joints infected for less than two weeks . The histological changes were more severe in infected arthritic joints than in infected normal joints (mean +/- SD total histology score, 13.8 +/- 2.4 and 9.3 +/- 4.0, respectively; P less than .001) . In infected arthritic joints, subsynovial abscesses extended into subchondral bone via the pannus of chronic synovitis at articular margins and intraarticular attachments of cruciate ligaments, rather than by initial cartilage destruction and direct extension into subchondral bone.

Endocrinol Jpn, 1986 Aug, 33(4), 489 - 96
Analysis of thyroglobulin antibody synthesis by cultured peripheral blood lymphocytes from patients with Hashimoto's thyroiditis using biotin-avidin solid phase enzyme immuno-assay; Hirose W et al.; The influence of bovine thyroglobulin (Tg) and/or staphylococcus aureus cowan I (SAC) on Tg antibody synthesis has been studied using cultures of 8 Hashimoto's and 5 normal peripheral blood lymphocytes (PBL) . The detection of Tg antibody in the culture supernatants was performed by sensitive biotin-avidin solid phase enzyme immunoassay . By using this technique, we were able to detect small amounts of Tg antibody synthesized by cultured Hashimoto's PBL responsive to bovine Tg and/or SAC; PBL from three out of eight patients produced increased levels of Tg antibody in the presence of 0.02 microgram/ml bovine Tg . On the other hand, PBL from two other cases among them which were unresponsive to bovine Tg alone became responsive to bovine Tg following simultaneous stimulation with SAC . PBL from the other three cases failed to respond to bovine Tg or simultaneous stimulation with bovine Tg and SAC . The former five patients had serum Tg tanned red cell hemagglutination (TGHA) titers greater than 1:409,600 except in one case and the latter had serum TGHA titers less than 1:12,800 . These results indicated the presence of the different functional stages of B cells to produce Tg antibody in the circulation of Hashimoto's patients and suggested that sufficient number of lymphocytes responsive to bovine Tg are present in the circulation of Hashimoto's patients with high titers of serum TGHA.

Chemioterapia, 1986 Aug, 5(4), 257 - 62
In vitro and in vivo evaluation of L/105, a new topical intestinal rifamycin; Venturini AP et al.; L/105 (4-deoxy-4'-methylpyrido {1',2'-1,2} imidazo {5,4-c} rifamycin SV; INN: Rifaximin) is a new rifamycin active in vitro against both gram-positive and gram-negative microorganisms . The activity of L/105 was comparable to that of rifampicin and, against gram-positive bacteria, higher than that of neomycin . The antibacterial activities of L/105 and rifampicin were equally affected by the highest size of inoculum used (10 cells/ml) and they were equally bactericidal against Staphylococcus aureus and Escherichia coli . The speed and the degree of development of resistance to L/105 were quite superimposable on those of neomycin . In vivo, L/105 did not show therapeutic activity by oral route in the staphylococcal infection in the mouse till the highest dosage used (10 mg/kg b.w.); under the same conditions, gentamicin was equally ineffective . After subcutaneous administration, L/105 showed therapeutic activity (ED50 = 0.46 mg/kg b.w.) practically superimposable on that of orally administered rifampicin.

South Med J, 1986 Aug, 79(8), 947 - 51
Chronic osteomyelitis caused by Staphylococcus aureus: controlled clinical trial of nafcillin therapy and nafcillin-rifampin therapy; Norden CW et al.; A controlled trial of treatment of chronic osteomyelitis caused by Staphylococcus aureus compared nafcillin alone with nafcillin plus rifampin for a six-week period . Treatment was well tolerated, the only adverse effect being mild neutropenia in four of 18 patients; no toxicity was observed from rifampin . Eight of ten patients in the combined treatment group had a favorable clinical response (with follow-up of two to four years) as compared to four of eight in the nafcillin group (P = .2) . Despite the failure to show a statistically significant advantage of rifampin plus nafcillin, we conclude that the combination, along with appropriate surgery, should be considered for patients with chronic staphylococcal osteomyelitis.

Biochem J, 1986 Aug 1, 237(3), 723 - 30
Reversible deactivation of beta-lactamase by quinacillin . Extent of the conformational change in the isolated transitory complex; Persaud KC et al.; Conditions have been established where the deactivation of the beta-lactamase from Staphylococcus aureus PC1 by the penicillin substrate, quinacillin, is close to complete but fully reversible . The temperature-dependence of the rate of re-activation indicated a half-life of about 170 min for the deactivated state at 0 degrees C . Measurement of the relative viscosity of mixtures of enzyme and quinacillin at 8.4 degrees C ruled out any significant difference in shape or solvation between the deactivated and the normal enzyme . C.d . measurements of the deactivated protein, separated from excess quinacillin, showed that the quinacillin side-chain chromophore was bound in an asymmetric environment . The ellipticity associated with the bound quinacillin chromophore decreased with the same first-order rate constant as that for reappearance of enzyme activity . These findings support the accumulation of a deactivated state that contains bound quinacillin or a derivative . Quinacillin caused a 3-fold increase in the rate of 3H exchange-out (at a rate that was low compared with that for the substantially unfolded or expanded protein) . However, there was rapid exchange-out of about 50 3H atoms on addition of 1 M-urea to the deactivated enzyme, whereas the same concentration had no effect on the exchange-out of 3H from native enzyme . The interpretation that quinacillin increases the susceptibility of the native state to unfolding in the presence of urea is supported by the demonstration that SO4(2)- ions decreased the rate and extent of deactivation but had no effect on the rate of re-activation, as predicted from the observation that SO4(2)- ions, in competition with urea, stabilize the native state relative to the partially unfolded state H {Mitchinson & Pain (1985) J . Mol . Biol . 184, 331-342}.

Clin Exp Immunol, 1986 Aug, 65(2), 303 - 10
In vitro synthesis of IgM rheumatoid factor by lymphocytes from patients with essential mixed cryoglobulinemia; Meroni PL et al.; Peripheral blood mononuclear cells (PBMC) from patients with Essential Mixed Cryoglobulinemia (EMC) were studied for their ability to synthesize polyclonal IgM and rheumatoid factor (RF) IgM in vitro . Our results indicate: that EMC-PBMC produce smaller amounts of polyclonal IgM but higher quantities of IgM-RF than normal PBMC after pokeweed mitogen (PWM) or Staphylococcus aureus activation, so that the IgM-RF to total IgM ratio is significantly greater in EMC than in normal cultures; that enriched EMC-B lymphocytes display a significantly higher spontaneous synthesis of IgM-RF than normal B lymphocytes and that the IgM-RF B cell clones are receptive to T cell regulation . Taken together these findings suggest an expansion of B cell clones committed to IgM RF production and the presence in peripheral blood of differentiated B lymphocytes capable of secreting IgM-RF in EMC.

Hum Genet, 1986 Aug, 73(4), 346 - 9
A subpopulation of t(2;14)(p11;q32) cells in ataxia telangiectasia B lymphocytes; Butterworth SV et al.; Partially purified B cells from ataxia telangiectasia (A-T) patients and normal individuals were stimulated with Staphylococcus aureus Cowan I organisms (SAC) . High levels of apparently random rearrangements were seen in the A-T B cells only . In addition a t(2;14)(p11;q32) rearrangement was identified in B cells from more than one patient.

J Infect Dis, 1986 Aug, 154(2), 283 - 8
Use of technetium-99m methylene diphosphonate and gallium-67 citrate scans after intraarticular injection of Staphylococcus aureus into knee joints of rabbits with chronic antigen-induced arthritis; Mahowald ML et al.; Numerous clinical studies have questioned the ability of radionuclide scans to differentiate septic from aseptic joint inflammation . A clinical study may not be able to document an underlying disease process or duration of infection and, thus, may make conclusions about the accuracy of scan interpretations open to debate . In this study, the Dumonde-Glynn model of antigen-induced arthritis in rabbits was used as the experimental model to study technetium and gallium scans in Staphylococcus aureus infection of arthritic and normal joints . Gallium scans were negative in normal rabbits, usually negative in antigen-induced arthritis, but positive in septic arthritis . The bone scan was usually negative in early infection but positive in late septic arthritis, a finding reflecting greater penetration of bacteria into subchondral bone because of the underlying inflammatory process.

J Oral Pathol, 1986 Aug, 15(7), 386 - 8
A comparison of oral rinse and imprint sampling techniques for the detection of yeast, coliform and Staphylococcus aureus carriage in the oral cavity; Samaranayake LP et al.; The sensitivity of the impression culture, the neat rinse culture (NRC) and the concentrated rinse culture (CRC) methods in detecting the oral carriage of yeasts, coliforms and Staphylococcus aureus was estimated in 75 individuals . The recovery of organisms from the imprint cultures of the tongue and the CRC was similar and there was highly significant positive correlation between the two techniques . The CRC was simple to perform, equally sensitive and superior in quantifying yeast, coliform and S . aureus carriage than the imprint culture technique . Hence, it is suggested that the CRC technique be preferentially employed in future investigations to obtain comparable data from different centres.

Allergy, 1986 Aug, 41(6), 423 - 8
Topical sodium cromoglycate in atopic dermatitis . A disappointing but informative trial; Kjellman NI et al.; Forty children with atopic eczema requiring topical steroids entered a double-blind group comparative study over 12 weeks and were randomized to either 4% sodium cromoglycate (SCG) in an oil-in-water cream or matching placebo cream . The eczema was evaluated on area charts for 20 parts of the body at five clinic visits . In addition, the families kept diaries on symptoms and treatment . After 3 weeks there were small but statistically significant decreases in severity scores recorded at the clinical visits in the SCG group compared with small increases in the placebo group . However, there were no statistically significant differences in the diary card data during the first 3 weeks of treatment or in any other period, nor were significant differences found in any efficacy data collected during the other 9 weeks of the trial . There were no marked differences in treatment opinions, unusual symptoms, skin infections, use of topical steroids or drugs, or acceptability data between the groups . Staphylococcus aureus was found once or twice in cultures from eczema lesions in 31 of 40 children with no marked group difference . The trial showed that there is great need for improved information, family support and topical as well as general treatment in childhood atopic eczema, but topical SCG did not relieve the patients' eczema.

J Interferon Res, 1986 Aug, 6(4), 331 - 6
Determination of the complete amino acid sequence of recombinant human gamma-interferon produced in Escherichia coli; Yamazaki S et al.; The complete amino acid sequence of recombinant human gamma-interferon (HuIFN-gamma) produced in Escherichia coli was determined using a gas-phase protein sequencer . The sequence was established by automated Edman degradation on the intact protein and its peptides obtained after Staphylococcus aureus V8 protease or trypsin digestion . The result was identical to the amino acid sequence predicted from the nucleotide sequence of the cloned HuIFN-gamma cDNA except that it was missing the four carboxy-terminal residues, Arg-Ala-Ser-Gln.

J Clin Microbiol, 1986 Aug, 24(2), 186 - 8
Screening method for recovery of methicillin-resistant Staphylococcus aureus from primary plates; La Zonby JG et al.; A study designed to screen for the presence of methicillin-resistant Staphylococcus aureus from primary plates was conducted from 1 January to 1 September 1985 in a small community hospital . The screening method used a plate of lipovitellin salt mannitol agar and a 4-microgram oxacillin disk incubated at 30 degrees C . Growth of yellow colonies, typical of S . aureus, around the disk without a zone of inhibition was called presumptive methicillin-resistant S . aureus . All susceptibilities were confirmed by using the National Committee for Clinical Laboratory Standards macrodilution technique . Of 224 cultures containing S . aureus, 118 (53%) were positive for methicillin-resistant S . aureus isolates . Of these 118, 111 (94%) were correctly identified from the primary plates as methicillin-resistant S . aureus . Of the 224 isolates, 14 could not be categorized from the primary plates as methicillin-resistant S . aureus due to the small amounts of S . aureus recovered.

Eur J Immunol, 1986 Aug, 16(8), 925 - 32
The roles of interleukin 2 and interferon-gamma in human B cell activation, growth and differentiation; Jelinek DF et al.; The roles of interleukin 2 (IL 2) and interferon-gamma (IFN-gamma) in human peripheral blood B cell activation, growth and differentiation were examined . Highly purified B cells stimulated with Cowan I Staphylococcus aureus (SA) proliferated minimally and generated no immunoglobulin-secreting cells (ISC) without the addition of T cell supernatants (T sup) produced by mitogen-activated T cells . Recombinant IL 2 (rIL 2) alone was able to promote maximum proliferation and generation of ISC in cultures of highly purified SA-stimulated B cells when present from the initiation of the incubation . IFN-gamma, by contrast, could not support either response alone . When a two-step culture system was employed to determine the effect(s) of T cell influences during both initial activation and in propagating the response following activation, it was found that B cells activated by SA alone subsequently responded maximally to T sup but only minimally to IL 2 and not at all to IFN-gamma . However, the presence of T sup, rIL 2, or rIFN-gamma during initial activation with SA was found to facilitate greatly the subsequent capacity of the activated B cells to proliferate and differentiate in response to either T sup or IL 2 . These data suggest two distinct pathways of human B cell responsiveness . Activities in T sup other than IL 2 or IFN-gamma can support the growth and differentiation of B cells initially activated with SA alone, whereas rIL 2 is capable of promoting these responses maximally only when B cells have been initially activated by SA in the presence of T cell lymphokines, such as IL 2 or IFN-gamma . The results emphasize the role of specific T cell factors in determining the outcome of humoral immune responses.

Proc Natl Acad Sci U S A, 1986 Aug, 83(16), 6174 - 8
Calcium and calmodulin-enhanced in vitro phosphorylation of hen brain cold-stable microtubules and spinal cord neurofilament triplet proteins after a single oral dose of tri-o-cresyl phosphate; Suwita E et al.; The effect of a single 750-mg/kg oral dose of tri-o-cresyl phosphate (TOCP) on the endogenous phosphorylation of brain microtubule preparations and spinal cord neurofilaments was assessed in hens after the development of delayed neurotoxicity . Protein phosphorylation with {gamma-32P}ATP was analyzed by one-dimensional and two-dimensional gel electrophoresis, autoradiography, and microdensitometry . TOCP treatment enhanced the Ca2+- and calmodulin-dependent phosphorylation of tubulin in crude chicken brain cytosol (160% for alpha-tubulin and 140% for beta-tubulin) and cold-stable microtubules (165% and 155% for alpha- and beta-tubulin, respectively) . Microtubule-associated protein 2 (MAP-2) phosphorylation was also increased in brain fractions studied--i.e., brain cytosol (145%), cold-stable microtubules (133%), and cold-labile microtubules (328%) . There was significant increase in phosphorylation of a 70-kDa protein in the brain cytosol and in the cold-stable microtubule fractions . TOCP also stimulated the phosphorylation of spinal cord proteins of 70 kDa (119%) and 160 kDa (129%) in a Mg2+-dependent manner . Addition of Ca2+ and calmodulin further enhanced the phosphorylation of these 70-kDa (563%) and 160-kDa (221%) proteins as well as of 52-, 59-, and 210-kDa proteins by as much as 126%, 160%, and 196%, respectively . Two-dimensional electrophoresis was carried out to identify these proteins . They were confirmed as alpha- and beta-tubulin (52 and 59 kDa) in brain and spinal cord preparations and the neurofilament triplet proteins (70, 160, and 210 kDa) in the spinal cord preparation . The 70-kDa protein in brain was not neurofilament in origin . Peptide mapping using Staphylococcus aureus V8 protease showed the brain and spinal cord cytoskeletal proteins have identical phosphopeptide patterns in control and TOCP-treated hens, indicating that it was unlikely that the phosphorylation sites were altered by TOCP treatment.

Proc Natl Acad Sci U S A, 1986 Aug, 83(15), 5439 - 43
Exchange of guanine nucleotide between GTP-binding proteins that regulate neuronal adenylate cyclase; Hatta S et al.; GTP-binding proteins have been demonstrated to stimulate and inhibit rat brain adenylate cyclase without the prior addition of hormone . Exposure of rat cerebral cortex membranes to hydrolysis-resistant GTP analogs results in inhibition (or stimulation) of adenylate cyclase, which persists subsequent to buffer washing . The hydrolysis-resistant GTP photoaffinity probe P3-(4-azidoanilido)-P1-5' GTP (AAGTP) can promote a similar persistent inhibition of adenylate cyclase, and, after removal of unbound AAGTP and subsequent UV photolysis, AAGTP is covalently linked to the 40-kDa inhibitory GTP binding protein, GNi (inhibitory guanine nucleotide binding regulatory subunit of adenylate cyclase) . Under conditions where the persistent inhibition of adenylate cyclase is overcome by subsequent incubation with 5'-guanylyl imidodiphosphate or NaF, AAGTP bound to the 40-kDa GNi protein is diminished while that bound to the 42-kDa stimulatory GTP-binding protein (GNs) is increased . Additionally, we have identified a 32-kDa protein that binds AAGTP with an affinity similar to that of GNs . This protein does not appear to be a byproduct of proteolysis as demonstrated by Staphylococcus aureus V8 protease digestion experiments, and it is not a substrate for ADP-ribosylation by bacterial toxins . The sum of the AAGTP bound by the GNi and GNs proteins is constant, and the transfer of nonphotoactivated AAGTP to GNs from GNi is stable to buffer washing . Furthermore, this alteration in the AAGTP-labeling pattern corresponds to the shift in adenylate cyclase from inhibition to stimulation . These data raise the possibility that hydrolysis-resistant GTP analogs might be exchanged directly between the GNi and GNs and that there exists some interaction between those proteins in the regulation of adenylate cyclase activity.

Int J Radiat Biol Relat Stud Phys Chem Med, 1986 Aug, 50(2), 337 - 43
Rhodium(III) as a potentiator of the effects of X-rays on cells; Richmond RC et al.; A rhodium compound, Rh(NH3)3Cl3, does not sensitize the spores of Bacillus megaterium to X-rays . However, it is a very effective sensitizer of vegetative cells of Staphylococcus aureus, raising the sensitivity four times in O2 and over 100 times in anoxia . The inhibition by oxygen of the sensitizing action of Rh(III), which operates over a wide range of {O2}, is noteworthy . These experiments were performed in saline-phosphate buffer using 50 kVp X-rays . The results are discussed in terms of the known radiation chemistry of this compound.

Aust J Exp Biol Med Sci, 1986 Aug, 64 ( Pt 4), 367 - 79
Conjugative, staphylococcal plasmids carrying hitch-hiking transposons similar to Tn554: intra- and interspecies dissemination of erythromycin resistance; Townsend DE et al.; Two staphylococcal plasmids, pWG4 and pWG25, encode production of a diffusible pigment and resistance to erythromycin and spectinomycin . The former was found occurring naturally in a clinical isolate of Staphylococcus aureus and the latter in S . epidermidis . Both plasmids are conjugative, capable of high-frequency, interspecies transfer, only isolated in the open-circular form and identical in molecular weight and pattern of restriction-endonuclease fragments . The only difference between the plasmids is in the expression of resistance, pWG4 encoding inducible and pWG25 constitutive erythromycin resistance . The resistance determinants of both plasmids behave as hitch-hiking transposons in cultural conditions that favour phage-mediated or phage-independent conjugation, always inserting a copy of themselves into the recipient's chromosome, except in S . epidermidis in which the chromosomal insertion site may be absent . The resistance determinants have been cloned and located on a 4 X 7 kbp EcoR1/HindIII restriction fragment which has a restriction map similar to that of the right arm of Tn554 (Murphy and Lofdahl, 1984) . The hitch-hiking transposon of plasmid pWG25 has been designated Tn3853.

Mol Gen Genet, 1986 Aug, 204(2), 341 - 8
Incompatibility between plasmids with independent copy control; Projan SJ et al.; Incompatibility between autonomous plasmids has been attributed, for the most part, to interaction between plasmids' negative control systems and/or partitioning systems . In this report it is shown that indirectly regulated plasmids with non-interactive negative control systems are incompatible on the basis of their shared initiator protein . This principle was demonstrated for a family of Staphylococcus aureus plasmids whose copy number is regulated by inhibitory RNAs that control the production of a rate-limiting, trans-active, initiator protein . We have constructed a pair of plasmids that have the same regulation systems and different initiator proteins and another pair with different regulation systems and the same initiators . Both of these pairs of plasmids were shown to be incompatible.

J Antimicrob Chemother, 1986 Aug, 18(2), 233 - 7
Effects of cefotaxime and cefodizime on human granulocyte functions in vitro; Labro MT et al.; In vitro, cefotaxime and cefodizime enhanced significantly the bactericidal activity of human neutrophils against Staphylococcus aureus P 209 A, but not phagocytosis . The increase was about 150% for cefotaxime and 400% for cefodizime at concentrations as low as 1 mg/l . Furthermore, by two different techniques (NBT and cytochrome C reduction tests) cefotaxime but not cefodizime significantly enhanced superoxide anion production by zymosan-stimulated neutrophils . Other neutrophil functions (chemotaxis and myeloperoxidase-mediated iodination of proteins) were not significantly altered by either antibiotic, even at concentrations as high as 1000 mg/l.

Proc Natl Acad Sci U S A, 1986 Aug, 83(16), 5793 - 7
Insulin stimulates the generation from hepatic plasma membranes of modulators derived from an inositol glycolipid; Saltiel AR et al.; Insulin binding to plasma membrane receptors results in the generation of substances that acutely mimic the actions of the hormone on certain target enzymes . Two such substances, which modulate the activity of the high-affinity cAMP phosphodiesterase (EC 3.1.4.17), have been purified from hepatic plasma membranes . The two have similar properties and activities but can be resolved by ion-exchange chromatography and high-voltage electrophoresis . They exhibit a net negative charge, even at pH 1.9, and an apparent molecular weight of approximately 1400 . The generation of these substances from membranes by insulin can be reproduced by addition of a phosphatidylinositol-specific phospholipase C purified from Staphylococcus aureus . This enzyme is known to selectively hydrolyze phosphatidylinositol and release from membranes several proteins that are covalently linked to phosphatidylinositol by a glycan anchor . Both enzyme-modulating substances appear to be generated by the phosphodiesterase cleavage of a phosphatidylinositol-containing glycolipid precursor that has been characterized by thin-layer chromatography . Some of the chemical properties of these substances have been examined . They appear to be related complex carbohydrate-phosphate substances containing glucosamine and inositol . These findings suggest that insulin may activate a selective phospholipase activity that hydrolyzes a membrane phospholipid, releasing a carbohydrate-containing molecule that regulates cAMP phosphodiesterase and perhaps other insulin-sensitive enzymes.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Aug, (8), 52 - 6
{Protective role of effectors and T-suppressors of delayed hypersensitivity in mice with a localized staphylococcal infection}; Bekhalo VA et al.; To induce delayed hypersensitivity (DH) in mice, experimental local infection with a small dose of Staphylococcus aureus was used . The production of suppressor cells was shown to occur after the intravenous injection of a large dose of killed staphylococcal culture . Experiments with the use of cell transfer and the treatment of lymphocytes with Thy-1 antiserum in the presence of the complement demonstrated the T-lymphocytic nature of DH and its suppression . The study revealed that the role played by DH in antistaphylococcal immunity was different in the animals infected by the subcutaneous routes; besides, the regulatory action of T-suppressors of DH was established.

EMBO J, 1986 Aug, 5(8), 1783 - 90
Inhibition of N-linked oligosaccharide trimming mannosidases blocks human B cell development; Tulp A et al.; Deoxymannojirimycin (dMM) or swainsonine (SW), which block conversion of high-mannose to complex-type N-linked glycans, strongly inhibited the production of immunoglobulin (Ig) when added to cultures of human lymphocytes together with the polyclonal B cell activators pokeweed mitogen (PWM) and Staphylococcus aureus (SAC) . To obtain the inhibitory effect, inhibitor had to be present during the first 36 h of culture . Addition at later timepoints was less effective and showed that neither inhibitor interfered with rate of production or secretion of Ig as such . Viability and proliferation of the lymphocytes, as defined by cell number and rate of DNA synthesis, were not influenced by the presence of dMM or SW, and no changes in the relative number of helper (T4+) or suppressor (T8+) cells were observed . Thus, for normal differentiation of human B lymphocytes into Ig secreting (plasma) cells in response to PWM and SAC, conversion of high-mannose to complex N-linked glycans is essential.

Proc Natl Acad Sci U S A, 1986 Aug, 83(15), 5474 - 8
Integration of staphylococcal phage L54a occurs by site-specific recombination: structural analysis of the attachment sites; Lee CY et al.; Lysogenization by staphylococcal phage L54a induces the loss of lipase (glycerol ester hydrolase) activity in its host Staphylococcus aureus . The attachment site of the bacterial chromosome (attB) for the phage is at the 3' end of the lipase gene, geh . The DNA fragment containing the attB (base pairs 2620-2637 inclusive) site has been sequenced . We have also cloned and determined the nucleotide sequence of the DNA fragments containing the other three attachment sites--i.e., the attP locus on the circularly permuted phage genome and the attL and attR loci at the left and right ends of the prophage in the lysogenized strain . These results reveal that an 18-base-pair core sequence is common to all four att sites . These data indicate that the crossover point must exist within the core sequence and, further, that integration is site- and orientation-specific . We also localized the viral recombinase gene to a 2.1-kilobase DNA segment extending rightward to the attP site . This region was found to be essential for integration of plasmids containing the attP site.

J Am Acad Dermatol, 1986 Aug, 15(2 Pt 2), 385 - 9
Staphylococcal scalded skin syndrome in a homosexual adult; Richard M et al.; A homosexual man developed staphylococcal scalded skin syndrome associated with a Staphylococcus aureus septicemia . We discuss the role of prednisone, renal insufficiency, and immunosuppression as predisposing factors to staphylococcal scalded skin syndrome in adults . In particular, our study of the patient's immune function revealed anergy and lymphopenia, with a reduced response to phytohemagglutinin . Studies of T cell subpopulations revealed an elevated percentage of T suppressor cells and a diminished percentage of T helper cells with a depressed T helper/T suppressor ratio . Because of those abnormalities, we suspected acquired immunodeficiency syndrome . A few months after recuperation from the acute disease, the patient has had a normalization of the T helper/T suppressor ratio, but because of persistent polyadenopathy, hypergammaglobulinemia, and a negative sensitization to keyhole-limpet hemocyanin (KLH), we now consider the patient to have an acquired immunodeficiency syndrome-related complex.

Infect Immun, 1986 Aug, 53(2), 366 - 71
Humoral antibody response against Bacteroides gingivalis-specific antigen recognized by monoclonal antibody in adult periodontal patients; Miyoshi T et al.; An antigen recognized by monoclonal antibody specific for Bacteroides gingivalis was purified in the presence of 0.5% (wt/vol) beta-octyl-glucoside by immunoadsorbent column chromatography . The purified antigen was homogeneous as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining, and the pattern of SDS-PAGE agreed with that of immunoblotting . The molecule exhibited an apparent molecular weight of about 62,000 by SDS-PAGE . The antigen was sensitive to trypsin, Staphylococcus aureus V8 protease, DNase I and II, and heating, but insensitive to RNase and neuraminidase . By the enzyme-linked immunosorbent assay method, the purified antigen was not cross-reactive with rat polyclonal antibodies to each of several black-pigmented Bacteroides species, Fusobacterium nucleatum, Eikenella corrodens, and Actinobacillus actinomycetemcomitans . These results indicate that the purified antigen is specific for B . gingivalis . Humoral antibody titers in adult periodontal patients against the specific antigen were measured by the enzyme-linked immunosorbent assay . Serum immunoglobulin G antibody titers against the specific antigen in adult periodontal patients correlated significantly with the severity of periodontal disease . However, such significant correlation was not observed with serum immunoglobulin M antibody titers.

Immunology, 1986 Aug, 58(4), 583 - 9
Control of human B-lymphocyte replication . I . Characterization of novel activation states that precede the entry of G0 B cells into cycle; Walker L et al.; Tonsillar B lymphocytes of a particularly high buoyant density were prepared essentially free of contaminating monocytes and T cells . When exposed to anti-immunoglobulin, such cells initiated the hydrolysis of inositol phospholipids . This provides a postulated 'dual signal' for growth through the liberation of intracellular calcium stores and the activation of protein kinase C . Nevertheless, neither anti-immunoglobulin nor direct agonists of this bifurcating pathway (respectively, calcium ionophore and the phorbol ester TPA) were capable, when used alone, of driving cells out of G0 and into RNA synthesis . All three agents did, however, induce two activation antigens at the surface of G0 B cells, which included CD23, p45 and a lineage-unrestricted antigen identified by the monoclonal antibody BK.19.9 . Cells that had been exposed to calcium ionophore, but not those activated with either TPA or anti-immunoglobulin, revealed further change indicated by an increased accessibility of their native DNA for the intercalating dye acridine orange . Cells receiving full mitogenic signals in the form of Staphylococcus aureus Cowan Strain I (SAC) or a combination of TPA and calcium ionophore showed the same initial sequelae but continued to enter the cell cycle and progress through to DNA synthesis . The observations identify two phases in the early activation of human B cells, both in terms of various temporal events, and the signals required to promote each activation state . before entering the proliferative cycle . Thus, the exit of human B cells from G0 appears subject to multiple controls that precede those associated with G1 and later phases of the cell cycle.

Biochem Biophys Res Commun, 1986 Jul 31, 138(2), 611 - 7
The complete amino acid sequence of human brain-derived acidic fibroblast growth factor; Gimenez-Gallego G et al.; Acidic fibroblast growth factor is a potent mitogen for a variety of cells in culture, including vascular endothelial cells, and is angiogenic in vivo . The complete amino acid sequence of human brain-derived acidic fibroblast growth factor has been determined from amino terminal sequence analysis and carboxypeptidase A digestion of the whole protein and sequence analyses of peptides generated by tryptic, Staphylococcus aureus V8 protease and cyanogen bromide cleavages . A potential Asn-Gly-Ser glycosylation sequence is present in the human protein . The complete amino acid sequence is compared to that of the equivalent protein purified from bovine brain.

Biochemistry, 1986 Jul 29, 25(15), 4309 - 14
Amino acid sequence of a basic Agkistrodon halys blomhoffii phospholipase A2 . Possible role of NH2-terminal lysines in action on phospholipids of Escherichia coli; Forst S et al.; A basic (pI = 10.2) phospholipase A2 of the venom of the snake Agkistrodon halys blomhoffii is one of a few phospholipases A2 capable of hydrolyzing the phospholipids of Escherichia coli killed by a bactericidal protein purified from human or rabbit neutrophil granules . We have shown that modification of as many as 4 mol of lysine per mole of the phospholipase A2, either by carbamylation or by reductive methylation {Forst, S., Weiss, J., & Elsbach, P . (1982) J . Biol . Chem . 257, 14055-14057}, had no effect on catalytic activity toward extracted E . coli phospholipids or the phospholipids of autoclaved E . coli . In contrast, modification of 1 mol of lysine per mole of enzyme substantially reduced activity toward the phospholipids of E . coli killed by the neutrophil protein . To explore further the role of lysines in the function of this phospholipase A2, we determined the amino acid sequence of the enzyme and the incorporation of {14C}cyanate into individual lysines when, on average, 1 lysine per molecule of enzyme had been carbamylated . After incorporation of approximately 1 mol of {14C}cyanate per mole of protein, the phospholipase A2 was reduced, alkylated, and exhaustively carbamylated with unlabeled cyanate . The amino acid sequence was determined of the NH2-terminal 33 amino acids of the holoprotein and of peptides isolated after digestion with trypsin and Staphylococcus aureus V-8 protease . The protein contains 122 amino acid residues, 17 of which are lysines . The NH2-terminal region is unique among more than 30 phospholipases A2 previously sequenced because of its high content of basic residues (His-1, Arg-6, and Lys-7, -10, -11, and -15).(ABSTRACT TRUNCATED AT 250 WORDS)

Biochemistry, 1986 Jul 29, 25(15), 4431 - 7
Identification of amino acid residues photolabeled with 2-azido{alpha-32P}adenosine diphosphate in the beta subunit of beef heart mitochondrial F1-ATPase; Garin J et al.; When beef heart mitochondrial F1-ATPase is photoirradiated in the presence of 2-azido{alpha-32P}adenosine diphosphate, the beta subunit of the enzyme is preferentially photolabeled {Dalbon, P., Boulay, F., & Vignais, P . V . (1985) FEBS Lett . 180, 212-218} . The site of photolabeling of the beta subunit has been explored . After cyanogen bromide cleavage of the photolabeled beta subunit, only the peptide fragment extending from Gln-293 to Met-358 was found to be labeled . This peptide was isolated and digested by trypsin or Staphylococcus aureus V8 protease . Digestion by trypsin yielded four peptides, one of which spanned residues Ala-338-Arg-356 and contained all the bound radioactivity . When trypsin was replaced by V8 protease, a single peptide spanning residues Leu-342-Met-358 was labeled . Edman degradation of the two labeled peptides showed that radioactivity was localized on the following four amino acids: Leu-342, Ile-344, Tyr-345, and Pro-346.

J Biol Chem, 1986 Jul 25, 261(21), 9678 - 83
Isolation of an active-site peptide of lipoprotein lipase from bovine milk and determination of its amino acid sequence; Reddy MN et al.; Lipoprotein lipase from bovine milk reacted stoichiometrically with diisopropylphosphorofluoridate (DFP), an inactivator of serine esterases, resulting in the loss of enzymatic activity against triacylglycerols . The reaction obeyed first-order kinetics with a rate constant of 0.69 h-1 . In order to isolate the peptide containing the diisopropylphosphoryl moiety (DIP), partially purified lipoprotein lipase was covalently labeled with {3H}DFP, and the labeled protein was reduced, carboxymethylated, and further purified to about 90% homogeneity . Cyanogen bromide cleavage followed by gel filtration yielded a radioactive peptide of 6-8 kDa . This peptide was succinylated and then digested with Staphylococcus aureus V8 proteinase . From this digest, a peptide containing 0.95 mol of {3H} DIP/mol of peptide was isolated by gel-permeation chromatography followed by reverse-phase high performance liquid chromatography . Automated Edman degradation provided the following sequence: Ala-Ile-Gly-Ile-His-Trp-Gly-Gly- (DIP)Ser-Pro-Asn-Gln-Lys-Asn-Gly-Ala-Val-Phe-Ile-Asn-(Ser, Leu)-Glu . Analysis of the sequence for secondary structure suggests that the reactive serine of lipoprotein lipase is in a beta-turn, a structure similar to those of the active sites of most other serine proteinases . Lipoprotein lipase appears to share this secondary structure with other serine hydrolases despite significant differences in the primary structure of this domain.

N Engl J Med, 1986 Jul 10, 315(2), 91 - 6
Staphylococcus aureus nasal carriage and infection in patients on hemodialysis . Efficacy of antibiotic prophylaxis; Yu VL et al.; We conducted a five-year prospective controlled study of prophylaxis of Staphylococcus aureus nasal carriage and infection among patients in a hemodialysis unit . Carriers tended to have chronic colonization with a single phage type . S . aureus infections occurred significantly more frequently in carriers than in noncarriers and, in 93 percent of the infected carriers, were caused by the same phage type as that carried in the nares . Neither intravenous vancomycin nor topical bacitracin was found to be efficacious in eradicating nasal carriage . However, oral rifampin given for five days decreased S . aureus carriage over a one-month follow-up period, but within three months colonization of the nares recurred in most carriers, often with an S . aureus of the original phage type . Carriers were then randomly assigned to receive either rifampin or no prophylaxis . Rifampin was readministered at three-month intervals if culture of the anterior nares yielded S . aureus . Infections with S . aureus occurred significantly more frequently in carriers given no prophylaxis than in those given a full course of rifampin . S . aureus resistant to rifampin was isolated from the anterior nares of four patients, but these isolates were not implicated in any infections . The incidence of infection at the dialysis access site, skin, and soft tissue of patients on hemodialysis can be decreased by interventions directed at nasal carriage of S . aureus.

FEBS Lett, 1986 Jul 7, 202(2), 303 - 8
The complete primary structure of GTP:AMP phosphotransferase from beef heart mitochondria; Tomasselli AG et al.; To complete the amino acid sequence of GTP:AMP phosphotransferase (MgGTP + AMP in equilibrium with MgGDP + ADP) from beef heart mitochondria it was necessary to sequence an intermediate region of about 33 residues after position 102 {(1984) Eur . J . Biochem . 143, 331-339} and find a suitable overlap with the rest of the protein . The required peptides were obtained by cleaving the enzyme with endoproteinase Lys-C . One peptide, covering the region from residue 79 to 144, was sequenced up to residue 124 . Another peptide, extending from residue 79 to 169, was subcleaved with Staphylococcus aureus V8 protease and provided the fragment from residue 99 to 139 which was sequenced . Several other peptides from endoproteinase Lys-C cleavage were used to check large sections of the previously published sequence work . The complete sequence contains 225 amino acids and has an Mr of 25 469.

J Biol Chem, 1986 Jul 5, 261(19), 8836 - 41
Chicken liver H-protein, a component of the glycine cleavage system . Amino acid sequence and identification of the N epsilon-lipoyllysine residue; Fujiwara K et al.; The complete amino acid sequence of H-protein from chicken liver was determined by aligning peptides obtained by cyanogen bromide, endoproteinase Lys-C, Staphylococcus aureus V8 protease, and chymotrypsin cleavage together with the partial NH2- and COOH-terminal sequence of the intact protein . H-protein consists of 125 amino acids and a lipoic acid moiety linked to lysine 59 . The sequence is: (sequence in text) . The lysyl residue involved in lipoic acid attachment is indicated with an asterisk . The molecular weight including lipoic acid is calculated to be 13,883 . From the secondary structure predicted by the method of Chou and Fasman (Chou, P . Y., and Fasman, G . D . (1978) Adv . Enzymol . 47, 45-148) the lipoic acid binding region shows alpha-helical structure and is predicted to be an interior portion of the protein from the hydropathic profile according to Kyte and Doolittle (Kyte, J., and Doolittle, R . F . (1982) J . Mol . Biol . 157, 105-132).

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1986 Jul, 261(4), 407 - 10
Coagulase typing of Staphylococcus aureus isolated from animals; Kawano J et al.; Coagulase typing was performed on Staphylococcus aureus isolated from various animals . Of the 383 strains examined, 209 (54.6%) strains were typable and could be differentiated into 8 coagulase types . The remaining 174 strains were non-typable with antisera against the 8 known types of coagulases . This suggests that many strains of S . aureus of animal origin may possess unknown types of coagulases.

Biol Chem Hoppe Seyler, 1986 Jul, 367(7), 579 - 89
Guinea pig plasma murinoglobulin . Purification and some properties; Suzuki Y et al.; Murinoglobulin, a newly identified mouse plasma protein resembling alpha-macroglobulins {Saito, A . & Sinohara, H . (1985) J . Biol . Chem . 260, 775-781}, was also found in guinea pig plasma, and purified to homogeneity . Guinea pig murinoglobulin consisted of a single 180-kDa polypeptide chain containing about 18% carbohydrate . It inhibited the proteolytic activities of trypsin and thermolysin towards Remazol brilliant blue hide powder, but stimulated the amidolytic activities of trypsin and Staphylococcus aureus V8 protease towards small synthetic substrates . Heat treatment of murinoglobulin completely abolished the former activities, but partially retained the latter activities . The ability of guinea pig murinoglobulin to inhibit the proteolysis was much weaker than that of the mouse homologue . On interaction with trypsin, murinoglobulin underwent cleavage of one susceptible bond with concomitant unmasking of one thiol group . Methylamine treatment also released one thiol group per molecule.

Antimicrob Agents Chemother, 1986 Jul, 30(1), 42 - 5
Steady-state serum pharmacokinetics of novobiocin and rifampin alone and in combination; Drusano GL et al.; Because of the potential of novobiocin-rifampin for oral therapy of methicillin-resistant Staphylococcus aureus infection, we evaluated the pharmacokinetics of novobiocin and rifampin, alone and in combination, in a randomized, crossover, multiple-dose evaluation (500 mg of novobiocin and 300 mg of rifampin administered orally, twice a day, for 27 doses) in 10 volunteers . The half-lives of novobiocin and rifampin when administered alone were 5.85 +/- 1.20 and 1.46 +/- 0.30 h, respectively; when administered in combination, the half-lives were 2.66 +/- 0.65 and 1.43 +/- 0.29 h, respectively . This difference was significant for novobiocin . The area under the curve also differed significantly for novobiocin when administered in combination . No significant differences were seen in the maximum concentration of drug in serum, the time to maximum concentration of drug in serum, or both for either drug when single and combination therapy groups were compared . A change in clearance of novobiocin rather than a change in absorption is the more likely explanation for these findings . The mechanism remains to be elucidated . Nevertheless, the trough serum concentrations of both novobiocin and rifampin were in excess of the MIC for 90% of strains tested of methicillin-resistant S . aureus, even when coadministered.

JPEN J Parenter Enteral Nutr, 1986 Jul-Aug, 10(4), 431 - 4
In vitro contamination of "piggyback/heparin lock" assemblies: prevention of contamination with a closed, positive locking device (Click-Lock); Gibilisco PA et al.; Direct contact and airborne transmission are established modes of microbial contamination of standard intravenous (iv) assemblies such as piggyback and heparin lock . In this study, 60% of the standard iv assemblies inoculated with Staphylococcus aureus (S . aureus) at the barrel of their exposed needle grew these organisms when cultured in a Soy Casein Digest Broth (SCDB) . Also, 40 closed, positive locking iv assemblies (Click-Lock) were inoculated at possible contamination sites, and none of these assemblies grew S . aureus in a SCDB . These in vitro studies suggest that a closed, positive locking iv assembly such as the Click-Lock device may substantially reduce, and potentially prevent contamination of iv systems.

Hautarzt, 1986 Jul, 37(7), 410 - 2
{Toxic shock syndrome}; Marsch WC et al.; A 23-year-old woman developed mitigated toxic shock syndrome while using intravaginal tampons during menstruation . The Staphylococcus aureus strain isolated from the vaginal epithelium produced the responsible exotoxin (TSST-1).

Genetika, 1986 Jul, 22(7), 1081 - 92
{Nucleotide sequence and physical map of kanamycin-resistant plasmid pUB110 from Staphylococcus aureus}; Bashkirov VI et al.; The complete nucleotide sequence of Staphylococcus aureus plasmid pUB10 was determined . The sequence consists of 4545 b.p . and contains 64% A-T and 36% G-C pairs . pUB110 was found to contain four open reading frames, capable of coding for polypeptides having more than 80 amino acids . All the putative polypeptides are coded for by one DNA strand . The molecular weights of four putative polypeptides are (in kilodaltons): A-49.5; B-38.8; C-28.8 and D-9.5 . Polypeptide C is involved in kanamycin resistance . Polypeptide B is, possibly, involved in pUB110 replication . No role has yet been established for polypeptides A and D, since deletions in their coding sequences have no detectable effect on any properties of pUB110 plasmid.

Arthritis Rheum, 1986 Jul, 29(7), 910 - 2
Aseptic arthritis in a man with toxic shock syndrome; Gertner E et al.; Synovitis in toxic shock syndrome (TSS) is an unusual finding . A 31-year-old man presented with pain and swelling in both knees and was found to have TSS, secondary to a septic bursitis caused by Staphylococcus aureus . Immune complexes were not detected in serum or synovial fluid, and the S aureus was not recovered from the inflamed joints . Antibodies against the TSS toxin-1 were detected in serum and synovial fluid, but in lower levels than would be seen in a normal control serum . Complement studies implicated alternative pathway activation by a marked diminution in C3 levels compared with C4 levels, and by lower levels of factor B than would be found in other inflammatory joint diseases . The diagnostic dilemma posed by TSS in a man is discussed.

Tijdschr Diergeneeskd, 1986 Jul 1, 111(13), 639 - 42
{A hemorrhagic-anemic syndrome with dermatitis in broiler chickens}; Froyman R et al.; Eleven successive deaths of broiler chickens affected with dermatitis (Staphylococcus aureus) preceded by anaemia and muscular haemorrhages, are reported as occurring in Belgium . All of these cases could be traced to a single flock of broiler breeders . The depletion of lymphoid cells in the bursa, thymus and spleen was a striking feature . Infectious Bursal Disease virus as triggering or causative agent, could not be demonstrated . The pathological changes showed a marked resemblance to similar syndromes which were recently reported in broilers (1, 2, 3, 4).

Am Surg, 1986 Jul, 52(7), 398 - 401
Social, economic, and surgical anatomy of a drug-related abscess; Wallace JR et al.; The "letting of pus" has been a surgical triumph throughout medical history . This study was initiated to expand our knowledge of the etiologic, economic, and surgical aspects of an abscess . The records of 651 patients undergoing intraoperative incision and drainage of an abscess over a 12-month interval were analyzed . The abscesses were due to injection of illicit street drugs in 421 patients (64.7%), perirectal, pilonidal or sweat gland inflammation in 118 patients (18%), complications of diabetes in 22 patients (3.4%) and a prior operative procedure in only 18 patients (2.8%) . Fifty-three of the 421 patients with a drug-related abscess were randomly selected for an indepth review . Eighty-three per cent of the patients had positive cultures including 75 per cent with a single organism and 25 per cent with mixed flora . Staphylococcus aureus was present in 62 per cent of the cultures and 41 per cent of the isolates were methicillin resistant . The average length of hospitalization was 12.4 days with a range of 1 to 61 days . The average cost of hospitalization was $10,651 which increased to $24,383 if the patient had a mycotic aneurysm . The estimated annual cost of treatment of this sequela of injected illicit drugs was 6.9 million dollars in our hospital.

J Pharmacol Methods, 1986 Jul, 15(4), 335 - 46
Radioimmunoassays for fish tail neuropeptides: I . Development of assay and measurement of immunoreactive urotensin I in Catostomus commersoni brain, pituitary, and plasma; Suess U et al.; Antisera were raised in rabbits using highly purified urotensin I (UI) or UI (4-41) conjugated with high-molecular-weight proteins . Antiserum 2U4, at a final dilution of 1:300,000, gave 50% binding to iodinated UI (specific activity 40-120 microCi/micrograms) . Carp (Cyprinus carpio) and sucker (Catostomus commersoni) UI peptides cross-reacted completely with the antiserum . Results from cross-reactivity tests using various tryptic and Staphylococcus aureus protease fragments of UI indicated that the antigenic sites of the main antibody(ies) are directed against a part sequence in the C-terminal region of the peptide although antibodies with lower concentration or affinity, recognizing N-terminal region(s) of the peptide, are also present . No cross-reactivity was seen with insulin, secretin, vasoactive intestinal peptide or a structural UI-homolog, the ovine hypothalamic corticotropin-releasing factor, up to molar concentrations . Sauvagine, another structurally homologous peptide from frog skin, showed only 0.1% cross-reactivity . The urophysis-specific proteins, urophysins B and D, showed a low degree of cross-reactivity . Assays of serial dilutions of urophysis extracts from teleostean species yielded parallel displacement curves, except in the case of the pacific goby, Gillichthys mirabilis . Varying concentrations of immunoreactive UI were present in different regions of C . commersoni brain, spinal cord, and pituitary, and also in plasma . The measurement of 17.5 +/- 2.1 fmol UI/ml of plasma indicates that this assay may be used for the study of circulating UI peptide(s).

J Vasc Surg, 1986 Jul, 4(1), 16 - 21
Inhibition of bacterial adhesion by antibacterial surface pretreatment of vascular prostheses; Webb LX et al.; Polytetrafluoroethylene grafts were pretreated with oxacillin, with the cationic detergent benzalkonium, or with both substances, either at room temperature or at 90 degrees C . Inhibition zones ranging from 6.4 to 15.2 mm formed around all grafts incubated on Staphylococcus aureus-streaked agar plates except control disks and those treated with oxacillin . Treated grafts were exposed in vitro to S . aureus in high concentration, followed by distilled water lavage . The graft surface was then stained with ruthenium red to stain polysaccharides and studied by scanning and transmission electron microscopy . Colonization of the graft surface by adhesive bacteria was demonstrated in all cases, although it was less prevalent on grafts pretreated with benzalkonium bound at 90 degrees C.

J Exp Med, 1986 Jul 1, 164(1), 321 - 6
C-reactive protein is produced by a small number of normal human peripheral blood lymphocytes; Kuta AE et al.; Biosynthetic labeling with {35S}met and immunoprecipitation with anti-C-reactive protein (CRP) antibodies and Staphylococcus aureus indicate that cell surface CRP is produced by lymphocytes . The ability of anti-CRP to reduce NK activity, and the demonstration that 125I-anti-CRP-labeled PBL are found in low-density Percoll fractions associated with large granular lymphocyte (LGL) and NK activity suggest that S-CRP-bearing cells are NK effectors . The production of S-CRP by LGL supports this hypothesis . While lymphocytes were shown to synthesize S-CRP, monocytes produced no detectable S-CRP . The lymphocytes that produce S-CRP apparently do not secrete it; when lymphocyte culture supernatants were tested, no S-CRP was found . This is the first description of extrahepatic synthesis of CRP.

J Clin Microbiol, 1986 Jul, 24(1), 137 - 8
Gastrointestinal carriage of methicillin-resistant Staphylococcus aureus; Rimland D et al.; Nasal and rectal cultures were taken from all patients with methicillin-resistant Staphylococcus aureus identified on routine cultures obtained because of clinical indications . Of 117 patients studied over a 3-year period, 70 (60%) had rectal colonization and 62 (53%) had nasal colonization . Rectal colonization, probably reflecting gastrointestinal carriage, may be a source of transmission of methicillin-resistant S . aureus in hospitalized patients and may be difficult to eradicate.

J Bacteriol, 1986 Jul, 167(1), 77 - 81
Binding of collagen to Staphylococcus aureus Cowan 1; Speziale P et al.; Collagen binds to a receptor protein present on the surfaces of Staphylococcus aureus cells . Binding of 125I-labeled type II collagen to its bacterial receptor is reversible, and Scatchard plot analysis indicates the presence of one class of receptor that occurs on an average of 3 X 10(4) copies per cell and binds type II collagen with a Kd of 10(-7) M . Studies on the specificity of collagen cell binding indicate that the receptor does not recognize noncollagenous proteins but binds all of the different collagen types tested (types I to VI) . Furthermore, isolated collagen alpha chains and peptides generated by cyanogen bromide cleavage of type I collagen alpha chains are recognized by the receptor as indicated by the ability of these polypeptides to inhibit binding of 125I-labeled type II collagen to staphylococcal cells . Synthetic collagen analogs were tested as inhibitors of type II collagen binding to bacterial cells . The peptides (Pro-Gly-Pro)n, (Pro-Pro-Gly)10, and (Pro-OH-Pro-Gly)10 were recognized by the receptor, whereas the peptides (Pro-Ala-Gly)n and polyproline showed no inhibitory activity.

Antimicrob Agents Chemother, 1986 Jul, 30(1), 157 - 60
Penetration of cefuzoname into the cerebrospinal fluid of rabbits; Haruta T et al.; Concentrations of cefuzoname in cerebrospinal fluid (CSF) were determined in a total of 16 rabbits, 5 with healthy meninges, 5 with Staphylococcus aureus meningitis, and 6 with Escherichia coli meningitis . Mean percentages of the maximum concentration of the drug in CSF versus that in serum were 0.57, 3.37, and 4.40% for healthy rabbits, those with staphylococcal meningitis, and those with E . coli meningitis, respectively . The percentages of the area under the concentration-time curve of cefuzoname in CSF versus that in serum were, in the order of healthy group, staphylococcal meningitis group, and E . coli meningitis group, 0.61, 4.99, and 8.04% at 15 to 60 min, 1.44, 7.09, and 12.7% at 15 to 120 min, and 1.87, 8.07, and 15.8% at 15 to 180 min after administration, showing significant differences between the healthy and meningitis groups . All of the values in the E . coli meningitis group were greater than those of the staphylococcal meningitis group, but the differences were not significant . The ratios of the half-life of cefuzoname in CSF to that in serum were 2.10, 1.98, and 3.37 for the healthy, staphylococcal meningitis, and E . coli meningitis groups, respectively, with no significant difference between the three groups . Cefuzoname seems to be among the middle ranks of beta-lactam agents as far as penetration rate is concerned; however, when its potent antibacterial activity and broad spectrum are taken into account, the concentrations in CSF in patients with meningitis seem worth examining.

Hepatology, 1986 Jul-Aug, 6(4), 579 - 86
Rat hepatic bile acid sulfotransferase: identification of the catalytic polypeptide and evidence for polymeric forms in female rats; Collins RH et al.; A monoclonal antibody, PK1B, directed against rat liver bile acid sulfotransferase was used for the purification and characterization of the enzyme . Incubation of rat liver supernatant with the antibody followed by immunoprecipitation with Staphylococcus aureus cells demonstrated that PK1B reacted with 90% of the enzymatic activity present in the liver supernatant from female rats and 40 to 50% of the activity in male liver preparations . Immunoadsorption chromatography with PK1B bound to Sepharose isolated active enzyme which was purified greater than 75-fold . Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of this preparation in the presence of 2-mercaptoethanol demonstrated three polypeptides: Mr 29,500; 32,500, and 34,000 . Western blot analysis indicated that PK1B recognized an epitope which was found only on the Mr 29,500 polypeptide . Two-dimensional gel electrophoresis associated the enzymatic activity with this Mr 29,500 band . High-pressure liquid chromatographic analysis of immunopurified enzyme defined three distinct, enzymatically active protein populations: I (Mr 400,000 to 170,000); II (Mr 130,000), and III (Mr 43,000) . An Mr 29,500 polypeptide was the sole constituent of Peaks I and III and a major constituent of Peak II . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence and absence of 2-mercaptoethanol indicated that in Peak II, catalytically active Mr 29,500 protein is associated with the other two polypeptides by disulfide bonds . In contrast, Peak I consists of a polymer of Mr 29,500 polypeptide which is independent of disulfide interaction.

Ann Thorac Surg, 1986 Jul, 42(1), 113 - 7
Cardiac surgery in patients with end-stage renal disease; Zamora JL et al.; In a retrospective study we analyzed the clinical features of 85 patients with end-stage renal disease who underwent cardiac operation . Seventy-eight patients were from reports in the literature, and 7 were from our experience . The cardiac procedures were primarily valve replacements and aortocoronary bypass (ACB) operations . The indication for valve replacement was most commonly infective endocarditis (73%), affecting most frequently the aortic valve (68%) . The most common organism was Staphylococcus aureus, and there was a recent episode of angioaccess site infection in at least 17.5% of patients with documented endocarditis . The 30-day mortality was 57% for patients undergoing emergency valve replacement and only 3% for similar elective operations . Cumulative survival at 48 months was equal to that of the overall hemodialysis population not having cardiac operations . The mean age (50 years), male to female ratio (9:1), number of vessels bypassed per patient (2.4), and operative mortality for ACB were equal to those reported in comparable series of patients with normal renal function . Cumulative survival at 48 months for ACB patients was similar (60% versus 56%) to that of the overall hemodialysis population . Cardiac operations can be performed safely in patients with end-stage renal disease; the morbidity and mortality are similar to those encountered in patients with normal renal function . The long-term survival after cardiac procedures in patients with end-stage renal disease is similar to that reported for the overall hemodialysis population not having cardiac operations.

Am J Med, 1986 Jul, 81(1), 43 - 52
Relationship of staphylococcal tolerance, teichoic acid antibody, and serum bactericidal activity to therapeutic outcome in Staphylococcus aureus bacteremia; Rahal JJ Jr et al.; A randomized cooperative study of therapy for Staphylococcus aureus bacteremia was conducted in which nafcillin was given for four or six weeks to patients with clinical endocarditis and for two or four weeks to those without evidence of endocarditis . Eighty-four patients were enrolled, and 32 completed treatment, all of whom had bacteriologic cures . Three patients, treated for two weeks, had complications that were undetectable by assay of serum teichoic acid antibody . Data were insufficient to allow conclusions regarding the optimal duration of therapy for patients with or without endocarditis . However, the results suggest that neither clinical nor immunologic methods can reliably detect complications in patients treated for two weeks only . In addition, patients infected with tolerant organisms remained febrile longer than those infected with nontolerant strains but did not require additional antibiotics for cure . Peak serum bactericidal activity at a dilution of 1:8 or greater was present in all patients . Serum bactericidal activity of 1:8 prior to an antibiotic dose was not necessary for cure.

Infect Immun, 1986 Jul, 53(1), 192 - 8
Bactericidal action of eosinophils from normal human blood; Yazdanbakhsh M et al.; The ability of normal human eosinophils to ingest and kill Staphylococcus aureus and Escherichia coli was investigated and compared with the reactions shown by neutrophils from the same donors . The rate of phagocytosis of S . aureus by eosinophils was 50% of that shown by neutrophils . Unlike neutrophils, eosinophils were not able to kill ingested S . aureus at low bacterium/phagocyte ratios . The degree of S . aureus killing increased with increasing ratios, being equal to that of neutrophils when bacterium/phagocyte ratios of about 15 were used . This was probably due to a better triggering of the eosinophil oxidase system at high bacterium/phagocyte ratios . The early kinetics of the association of bacteria with eosinophils, the perforation of the bacterial envelope and the inactivation of bacterial proteins, was monitored in the ML-35 mutant strain of E . coli . The association of E . coli with eosinophils was 70% of that with neutrophils . Eosinophils had only 25% of the capacity of neutrophils to perforate the E . coli envelope . E . coli loses its colony-forming ability when the bacterial envelope has been perforated, indicating that eosinophils also kill E . coli more slowly than do neutrophils . This was confirmed with a plating assay for colony formation . The perforation of E . coli is independent of peroxidase-mediated reactions . Hence, the defective bactericidal action of eosinophils is probably not related to the differences between myeloperoxidase and eosinophil peroxidase . On the other hand, the inactivation of bacterial proteins is peroxidase dependent and was also seen to occur to a lesser extent in eosinophils compared with neutrophils . We conclude that eosinophils ingest E . coli but only slowly perforate (kill) these bacteria and barely inactivate the bacterial enzymes . In contrast, neutrophils quickly ingest and perforate (kill) E . coli and quickly inactivate the bacterial enzymes.

J Immunol, 1986 Jul 1, 137(1), 15 - 27
Contributions of the Mac-1 glycoprotein family to adherence-dependent granulocyte functions: structure-function assessments employing subunit-specific monoclonal antibodies; Anderson DC et al.; MAb directed at the alpha-subunits of Mac-1 (alpha M), LFA-1 (alpha L), p150,95 (alpha X), or their common beta-subunit were used to characterize the contributions of the Mac-1 glycoprotein family to granulocyte adherence reactions . Inhibitory effects of these MAb in incubation experiments with normal granulocytes indicated distinct adhesive contributions of each subunit . Significantly greater adherence, and inhibition of adherence by anti alpha M, alpha X, and beta MAb, was observed under chemotactic conditions designed to "up-regulate" the surface expression of the alpha M beta and alpha X beta complexes . Adherence to protein-coated glass and binding of albumin-coated latex beads were significantly inhibited by anti-beta greater than anti-alpha M (OKM-10, M1/70, LM2/1.6 and OKM-1) greater than anti-alpha X greater than anti-alpha L MAb, but no effects of anti-HLA, AB, or anti-CR-1 MAb were evident . A similar rank order of inhibition was observed in granulocyte aggregation assays in response to C5a, PMA, or f-Met-Leu-Phe . Significant inhibition of directed migration by anti-beta or anti-alpha M (OKM-1 or OKM-10) MAb was observed in subagarose but not Boyden chemotaxis assays; inhibition was dependent on a continuous cell exposure to anti-Mac-1 alpha or beta during the assay, suggesting that a continuum of new Mac-1 expression is required for directed translocation . Phagocytosis of Oil-Red-O paraffin or zymosan selectively opsonized with C3-derived ligands was significantly inhibited by anti-alpha M MAb (OKM-10 greater than LM2/1.6 greater than M1/70 greater than OKM-1) or by combinations of anti-alpha M + anti-CR-1 MAb, but only minimal inhibitory effects of anti-beta MAb and no effects of anti-alpha L or anti-alpha X MAb were seen . Similarly, complement-dependent phagocytosis-associated lactoferrin release, ingestion, and intracellular killing of Staphylococcus aureus 502A, and binding of iC3b-opsonized SRBC, were significantly inhibited by anti-alpha M (OKM-10, M1/70) or combinations of anti-alpha M + anti-CR-1 MAb, but not by anti-beta, alpha L, or alpha X MAb . Notably, none of the anti-Mac-1 MAb demonstrated inhibitory effects in assays of adherence-independent functions including shape change, specific f-Met-Leu-3H-Phe binding, O-2 generation, chemiluminescence evolution, or lactoferrin release in response to PMA.(ABSTRACT TRUNCATED AT 400 WORDS)

J Antimicrob Chemother, 1986 Jul, 18(1), 27 - 33
Relationship between penicillinase production and the in-vitro activity of methicillin, oxacillin, cloxacillin, dicloxacillin, flucloxacillin, and cephalothin against strains of Staphylococcus aureus of different phage patterns and penicillinase activity; Frimodt-Moller N et al.; A total of 157 strains of Staphylococcus aureus of different phage patterns and penicillinase production were investigated for their susceptibility to methicillin, oxacillin, cloxacillin, dicloxacillin, flucloxacillin and cephalothin by an agar dilution method . Only strains of the 52, 52A, 80, 81 complex had significantly higher IC-50 values than the rest of the strains . No correlation was found between penicillinase production and the IC-50 values . Penicillinase susceptibility divided the antibiotics into two groups: one including methicillin, oxacillin and cephalothin, and the other included dicloxacillin, cloxacillin and flucloxacillin . Nineteen strains of S . aureus which existed in both a penicillinase producing and a penicillinase non-producing form were examined for susceptibility to the six antibiotics . The difference between penicillinase positive and penicillinase negative variants was especially marked for flucloxacillin and cephalothin . Methicillin induction prior to susceptibility testing had only a minor influence on the results . Investigation of the stability of methicillin and the four isoxazolyl penicillins against penicillinase production by 37 strains of S . aureus showed methicillin to be the most stable antibiotic . This was followed by dicloxacillin, cloxacillin, flucloxacillin, and oxacillin in that order . The order of stability was identical and independent of phage pattern and quantitative penicillinase production.

Am J Clin Pathol, 1986 Jul, 86(1), 97 - 101
Presence of beta-lactamase-producing bacteria and beta-lactamase activity in abscesses; Brook I; The presence of beta-lactamase-producing bacteria (BLPB) in abscesses was investigated in 109 abscesses . Single isolates were recovered in 23 (21%) instances and were predominantly Staphylococcus aureus . The other abscesses yielded growth of two or more aerobic and/or anaerobic organisms . Aerobic bacteria were recovered in 28 (26%) of the aspirates, anaerobic bacteria in 41 (38%), and mixed aerobic and anaerobic bacteria in 40 (36%) . A total of 362 isolates (247 anaerobes and 115 aerobes) were recovered, accounting for 3.3 isolates per specimen (2.2 anaerobes and 1.1 aerobes) . One hundred beta-lactamase-producing organisms were recovered in 88 (77%) specimens . These included all 28 isolates of Bacteroides fragilis, 18 of 30 Bacteroides melaninogenicus, 42 of 43 S . aureus, and 11 of 14 Escherichia coli . Beta-lactamase activity was detected in 40 (55%) of the purulent specimens when using the chromogenic cephalosporin nitrocefin method . These data demonstrate the presence of aerobic and anaerobic BLPB in abscesses.

J Infect Dis, 1986 Jul, 154(1), 55 - 63
Induction of interleukin-1 by strains of Staphylococcus aureus from patients with nonmenstrual toxic shock syndrome; Parsonnet J et al.; We studied the induction of human interleukin-1 (IL-1) production in strains of Staphylococcus aureus isolated from patients with nonmenstrual toxic shock syndrome (TSS) . Of the 20 TSS-associated strains studied, 11 produced and nine did not produce TSS toxin-1 (TSST-1) . Human monocytes were incubated with dilute staphylococcal supernatants, and IL-1 production was measured in a lymphocyte-activating factor assay . All 20 TSS-associated strains were potent inducers of IL-1, in comparison with none of 10 vaginal isolates of S . aureus from healthy women . TSST-1-positive strains were more potent than TSST-1-negative strains . Nine TSST-1-negative TSS-associated strains were compared with 14 strains of S . aureus from other clinical settings and were found to be significantly more potent inducers of IL-1 (P less than .01) . Eight of these nine TSS-associated strains produced at least one staphylococcal enterotoxin . Stimulation of monocytes by products of S . aureus may play a role in the pathogenesis of TSS.

Arch Dermatol, 1986 Jul, 122(7), 815 - 7
Isotretinoin and Staphylococcus aureus infection . A possible association; Graham ML 2nd et al.; The use of isotretinoin (13-cis-retinoic acid) in the treatment of numerous dermatologic disorders, as well as the side effects encountered with use of the drug, have increased remarkably since its release . We encountered a case of Staphylococcus aureus endocarditis in a patient with chronic stable aortic insufficiency undergoing therapy with isotretinoin for extensive actinic keratoses . Although significant dysfunction of the immune system has not been demonstrated with isotretinoin, nasal colonization with S aureus has been shown to occur . Changes in skin fragility caused by the drug may provide a portal of entry for the organism . Physicians should be alert for this potential complication in patients with an underlying cardiac valvular lesion; antibiotic prophylaxis may be indicated in this group during isotretinoin therapy.

J Leukoc Biol, 1986 Jul, 40(1), 43 - 53
Staphylococcus aureus-derived chemoattractant activity for human monocytes; Rot A et al.; Products of bacteria are potent chemoattractants for mammalian leukocytes . Several reports suggest that these attractants are small peptides . We compared the properties of culture fluids of Staphylococcus aureus with fMet-Leu-Phe, considered a prototype of bacterial attractant . Chemotactic activity for human monocytes of Staph . aureus culture filtrates was determined in multiwell chemotaxis chambers . At optimal concentrations, the filtrate attracted almost twice as many monocytes as fMet-Leu-Phe (53 +/- 5% of input number compared with 30 +/- 3%, in a series of ten experiments) . Gel-filtration characteristics and susceptibility to proteolytic digestion suggested that chemotactic activity was due to peptides with a molecular size range of 500-2,000 daltons . Reverse-phase high-pressure liquid chromatography (HPLC) of unfractionated filtrate revealed nine peaks of chemotactic activity, most of which was in five of the peaks . One peak accounted for 40% of total activity . Individual peaks, like the unfractionated material, were capable of attracting about twice as many monocytes as the optimal concentration of fMet-Leu-Phe . Quantitative bioassay of the HPLC peaks showed that only 5% of the total Staph . aureus chemotactic activity eluted in the position of fMet-Leu-Phe . This is in contrast to the report that fMet-Leu-Phe accounted for 70% of neutrophil lysosomal enzyme-releasing activity of Escherichia coli culture fluid . In summary, chemotactic activity for monocytes of Staph . aureus culture fluid is due to peptides other than fMet-Leu-Phe; these peptides recruit a higher percentage of monocytes than fMet-Leu-Phe.

Orthop Rev, 1986 Jul, 15(7), 440 - 2
Septic arthritis of the sternoclavicular joint; Blankstein A et al.; An adult patient with isolated septic arthritis of the sternoclavicular joint (SCJ) caused by Staphylococcus aureus is described . The patient had no underlying diseases other than paronychia, which might have served as the portal of entry . Antibiotic therapy resulted in clinical cure but x-ray revealed progression of the destructive process.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1986 Jul, 182(4), 372 - 80
{Germicidal experiments with aqueous PVP-iodine-containing disinfecting solutions: effect of the content of free iodine on the bactericidal action against Staphylococcus aureus}; Gottardi W et al.; The bactericidal activity against Staph . aureus of three PVP-iodine preparations in different concentrations, and of 0.1 mol/L KI-solutions with concentrations of free iodine corresponding to the povidone-iodine preparations, was evaluated at reaction times of 7, 15, 30 and 60 seconds . The concentration of free iodine has been measured potentiometrically during the entire reaction time . In all cases it has been confirmed that the bactericidal activity is increasing with the concentration of free iodine . However, there have been found differing correlations between the logarithmic decrease of germs (RF-value), the concentration of free iodine and the reaction time (statistical evaluation of the results by means of linear multiple regression: RF = a0 + a1 log {I2} + a2 log t) . Referring to the concentration of free iodine the bactericidal activity in the range of RF approximately equal to 4-6 is increasing as follows: Povidone-iodine washing concentrate much greater than iodine in KI (0.1 mol/l) greater than Povidone-iodine mucosal disinfectant greater than aqueous solution of povidone iodine . This sequence is explained by differences of composition . So it is assumed that the surface active ingredients facilitate the penetration of molecular iodine . The observed correlations show that there is probably no exact mathematical relation of general validity for iodine preparations of different composition between the concentration of free iodine and the RF-value . However, for one and the same preparation such a correlation can be derived and enables to make predictions about the bactericidal activity, which can be expected as a consequence of the concentration of free iodine.

Pharmazie, 1986 Jul, 41(7), 475 - 8
Syntheses and in vitro antimicrobial evaluation of some benzimidazol-2-ylmethylthioureas, benzimidazol-2-ylacetylthiosemicarbazides and products of their condensation with monochloroacetic acid; Rida SM et al.; N-Benzimidazol-2-ylacetyl-N'-{alkyl- and arylthio (carbamoyl)}hydrazines and N-benzimidazol-2-ylmethyl-N'-alkyl- and -arylthioureas were subjected to condensation with monochloroacetic acid to afford N-benzimidazol-2-ylacetyl-N'-2,3, 4,5-tetrahydro-4-oxo-3-alkyl- and -arylthiazol-2-ylidenehydrazines and 3-benzimidazol-2-ylmethyl-2-alkyl- and arylimino-2,3-dihydrothiazol-4-(5H)ones, respectively . In preliminary antimicrobial testing, some compounds turned out to have significant activity against Staphylococcus aureus.

Am J Otolaryngol, 1986 Jul-Aug, 7(4), 298 - 301
Comparison of bacteria in the tympanic cavity and the mastoid antrum in chronic otitis media; Yamamoto E et al.; Materials collected from the tympanic cavity before operation and from the mastoid antrum during operation of 58 discharging ears of patients with chronic otitis media were cultured, and the bacteria in these two cavities were compared . Staphylococcus aureus was the commonest organism in the tympanic cavity, and S epidermidis in the mastoid antrum . Anaerobic bacteria were found only in the mastoid antrum of patient with cholesteatoma . In 32 (55 per cent) of the 58 ears examined, bacteria were detected in both the tympanic and mastoid cavities . In 17 ears (53 per cent), the bacterial strains in the two cavities differed . The results indicate the necessity of bacteriologic examination of the mastoid cavity during operation to select antibiotics for postoperative treatment.

Antimicrob Agents Chemother, 1986 Jul, 30(1), 137 - 42
Synergy between RU 28965 (roxithromycin) and human neutrophils for bactericidal activity in vitro; Labro MT et al.; The in vitro effects of RU 28965 (roxithromycin), a new semisynthetic macrolide, on human neutrophil activity were compared with those of erythromycin . RU 28965, at a concentration as low as 0.1 microgram/ml, significantly enhanced the phagocytosis and killing of Staphylococcus aureus by neutrophils . Erythromycin displayed a less stimulating effect in a dose-dependent manner . Phagocytosis of Klebsiella pneumoniae was also increased after incubation of neutrophils with RU 28965, but killing was not altered . Neutrophil chemotaxis, myeloperoxidase activity, and O2 consumption were unchanged in the presence of RU 28965.

J Clin Microbiol, 1986 Jul, 24(1), 131 - 6
Characterization of clinical strains of Staphylococcus aureus associated with pneumonia; Sanford BA et al.; A total of 5 Staphylococcus aureus strains from patients with postinfluenzal staphylococcal pneumonia, 7 from burn patients with staphylococcal pneumonia, and 21 from the nasopharynx of carriers were phenotypically characterized . All or most strains produced coagulase, clumping factor, DNase, thermostable DNase, protease, gelatinase, lipase, and pigment; the strains were low to moderate producers of extracellular protein A, fibrinolysin, and alpha-hemolysin . All strains were sensitive to mercury, half were sensitive to arsenate and cadmium, and 67 to 92% were resistant to penicillin . Differences between strains were not statistically significant . Cell surface hydrophobicity was determined by measuring percent adsorption to hexadecane . Hydrophobicity of postinfluenzal staphylococcal pneumonia strains was significantly lower than that of pneumonia strains from burn patients and carriers (P less than 0.005) . Immunoblot experiments with sera immune to one clinical test strain allowed the separation of all strains into three groups based on probe-positive reactions with primarily four staphylococcal polypeptides (154,200, 130,000, 77,100, and 64,400 molecular weight) . The difference in distribution of clinical and carrier strains was highly significant (P = 0.007).

Eur J Immunol, 1986 Jul, 16(7), 761 - 6
B cell growth and differentiation factors interact with receptors distinct from the interleukin 2 receptor; Kehrl JH et al.; Human B lymphocytes preactivated with Staphylococcus aureus Cowan strain I can proliferate and differentiate to Ig-secreting cells when cultured in the presence of recombinant interleukin 2 (IL2) . We have compared 2 different B cell growth factors (BCGF) and a B cell differentiation factor (BCDF) to IL2 in the regulation of human B lymphocyte growth and differentiation . Utilizing a competitive binding assay, neither a high molecular weight BCGF (HMW-BCGF) nor a low molecular weight BCGF (LMW-BCGF) competitively inhibited the binding of radiolabeled IL2 . Blocking studies with the anti-Tac monoclonal antibody demonstrated that B cell proliferation to IL2 was inhibited while a crude supernatant containing BCGF and IL2 was only partially inhibited . B cell Ig secretion induced by IL2 was also inhibited by anti-Tac while a crude supernatant and partially purified BCDF were not . Furthermore, IL2 plus BCGF was shown to enhance B cell proliferation better than either factor alone and IL2 plus a BCDF enhanced B cell Ig secretion better than either factor alone . By separating activated B cells into Tac-positive and Tac-negative fractions, much of the previously noted enhancement with the 2 factors was found to be secondary to the differential sensitivity of the 2 populations to BCGF and IL2 or BCDF and IL2 . Thus, LMW-BCGF, HMW-BCGF and a partially purified BCDF appear to interact with receptors distinct from the IL2 receptor in mediating their effects on B cell growth and differentiation.

J Antibiot (Tokyo), 1986 Jul, 39(7), 978 - 84
Susceptibility of bacteria to serum lysis or phagocytosis following growth in subinhibitory levels of lincosaminide or spectinomycin related antibiotics; Cialdella JI et al.; The effects of subinhibitory concentrations of antibiotics on polymorphonuclear leukocyte (PML) and serum killing of Staphylococcus aureus 502A (UC 9116) and Escherichia coli UC 9451 were studied . Exposure of these bacteria to subinhibitory levels of certain lincosaminides, spectinomycin, or 6'-n-propylspectinomycin altered their susceptibility to these host defense mechanisms, while exposure to gentamicin had no effect . However, each organism responded differently to treatment with the antibiotics . S . aureus pretreated during log phase growth with subinhibitory concentrations of clindamycin, lincomycin, or pirlimycin was more susceptible to killing by PMLs than untreated bacteria . No effect on phagocytic killing was found when S . aureus was pretreated with spectinomycin, 6'-n-propylspectinomycin, or gentamicin . The S . aureus remained resistant to serum lysis despite antibiotic treatment . In contrast, spectinomycin and 6'-n-propylspectinomycin as well as clindamycin dramatically increased the susceptibility of E . coli to serum lysis (greater than 99% destroyed) . Moderate killing of E . coli by PMLs was also found.

Antimicrob Agents Chemother, 1986 Jul, 30(1), 161 - 9
Conjugative transfer of staphylococcal antibiotic resistance markers in the absence of detectable plasmid DNA; el Solh N et al.; Eleven Staphylococcus aureus clinical isolates were tested for transfer of resistance markers by transduction and filter mating . The resistance markers of six of the strains could be transferred only by transduction; however, the five remaining strains transferred their resistance both by transduction and filter mating . The resistance markers that were cotransferred in filter matings (transfer of resistance to penicillin and streptogramin A was accompanied, in each case, by the transfer of one or more markers, i.e., resistance to aminoglycosides, cadmium, or tetracycline, depending on the donor) were not cotransduced . The filter mating transfers were recA independent and were observed with both Staphylococcus aureus and Staphylococcus epidermidis recipients . Experiments to elucidate the mechanism of transfer by filter mating suggested that conjugation requiring cell-to-cell contact may have been involved . These transfers occurred in the absence of detectable plasmid DNA.

J Vasc Surg, 1986 Jul, 4(1), 5 - 7
Rifampin protection against experimental graft sepsis; McDougal EG et al.; The risk of infection in vascular prosthetic conduits appears to be greatest in the perioperative period and the organism most frequently found is Staphylococcus aureus . Previous work suggests that antibiotics must be chemically bonded to the material to resist rapid washout caused by the flow of blood through the graft . The exception to this is rifampin, which remains fixed in Dacron prostheses after passive addition of the agent to aliquots of blood used to clot the interstices of porous Dacron grafts . This characteristic of rifampin is presumed to be caused by its poor water solubility . This potential infection resistance was challenged in a standard model of a canine infrarenal aortic graft by intravenous infusion of S . aureus organisms (10(7)) in the perioperative period . The grafts of five animals were preclotted with 9 ml of autogenous blood plus 1 ml of rifampin (60 mg/ml) . A second group had similar procedures with 1 ml of cefazolin (238 mg/ml) substituted for the rifampin, and a control group had 1 ml of saline solution added to the 9 ml aliquot of blood . The animals were killed at 3 weeks and examined for clinically apparent infection . Rings of the graft material were also removed aseptically and cultured . All five grafts preclotted with cefazolin had clinical and culture evidence of infection (S . aureus), as did the grafts of three of the five control dogs . None of the grafts preclotted with rifampin was infected (p less than 0.05) . Addition of rifampin to the blood used to clot the graft interstices appears to be a simple way of imparting graft resistance to perioperative sepsis.

Clin Immunol Immunopathol, 1986 Jul, 40(1), 50 - 61
Phagocyte defects; White CJ et al.; Although inherited forms of phagocyte defects affect a small proportion of the general population, their clinical course can be altered dramatically by a physician's awareness of these diseases and modifications of the approach to and treatment of affected patients . The most common syndromes are chronic granulomatous disease of childhood (CGD), the Chediak-Higashi syndrome (CHS), the hyperimmunoglobulin-E-recurrent infection (Job's) syndrome (HIE), and myeloperoxidase (MPO) deficiency . CGD patients have defects in the oxidative metabolism involved in killing catalase-positive organisms . CHS patients have giant granules defective in fusing with phagosomes and subsequent killing of ingested organisms . HIE patients have abnormal chemotaxis and elevated IgE levels and are susceptible to skin infections with Staphylococcus aureus and recurrent sinopulmonary infections . MPO-deficient patients often go undetected since they rarely have recurrent infections unless they have a concomitant disease such as diabetes mellitus . Patients with a recently described syndrome, C3bi receptor deficiency, have recurrent bacterial infections and persistent leukocytosis, and their neutrophils have abnormal adherence and phagocytosis . The absence of specific granules is a more rare entity but these patients also have recurrent infections thought to be secondary to a chemotactic defect and a minor abnormality of microbial killing exhibited by their neutrophils . This review will focus on the clinical presentation and management of these patients.

J Neurochem, 1986 Jul, 47(1), 254 - 62
Regulation of the actin-activated Mg-ATPase of brain myosin via phosphorylation by the brain Ca2+, calmodulin-dependent protein kinases; Tanaka E et al.; We have previously isolated two Ca2+, calmodulin-dependent protein kinases with molecular weights of 120,000 (120K enzyme) and 640,000 (640K enzyme), respectively, by gel filtration analysis from rat brain . Chicken gizzard myosin light-chain kinase and the 120K enzyme phosphorylated two light chains of brain myosin, whereas the 640K enzyme phosphorylated both the two light chains and the heavy chain . The phosphopeptides of the light chains digested by Staphylococcus aureus V8 protease were similar among chicken gizzard myosin light-chain kinase, the 120K enzyme, and the 640K enzyme . Only the seryl residue in the light chains and the heavy chain was phosphorylated by the enzymes . The phosphorylation of brain myosin by any of these enzymes led to an increase in actin-activated Mg-ATPase activity . The results suggest that brain myosin is regulated by brain Ca2+, calmodulin-dependent protein kinases in a similar but distinct mechanism in comparison with that of smooth muscle myosin.

J Hosp Infect, 1986 Jul, 8(1), 24 - 30
Osteomyelitis with methicillin-resistant Staphylococcus aureus; Fitzpatrick DJ et al.; We describe 10 patients with hospital-acquired osteomyelitis due to methicillin-resistant Staphylococcus aureus . The patients were posttraumatic and eight had a foreign body in situ at the site of infection . Vancomycin therapy in association with radical debridement was followed by clinical and radiological cure in eight patients at 2-3.5 years follow-up, in two of whom a foreign body was left in situ . Only minor adverse effects of vancomycin therapy (one rash, two thrombophlebitis) were seen.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Jul, (7), 48 - 51
{Range of the antigenic specificity of peptidoglycan in Staphylococcus aureus compared to other representatives of the Staphylococcus genus}; Astaf'ev DG et al.; In investigations based on the use of a highly sensitive test system permitting the detection of normal human antibodies to S . aureus peptidoglycan, the antigenic relationships between the peptidoglycans of S . aureus and other representatives of the genus Staphylococcus have been studied . Among other staphylococcal species, S . simulans, S . xylosus, S . hyicus, S . cohnii, S . hyicus s . s . chromogenes have been found to possess peptidoglycans most closely related to S . aureus peptidoglycans, while S . warneri and S . epidermidis peptidoglycans have proved to be least closely related to it.

Am J Med, 1986 Jun 30, 80(6B), 222 - 7
Antibiotics delivered by an implantable drug pump . A new application for treating osteomyelitis; Perry CR et al.; Osteomyelitis is usually treated with wide surgical debridement and prolonged intravenous antibiotics . The advent of the implantable drug pump has led us to evaluate this therapeutic modality for the treatment of osteomyelitis . We have previously shown that amikacin retains microbiologic activity in an implantable drug pump over a six-week period when incubated at 37 degrees C . We have also demonstrated in rabbits that high local levels and low systemic levels of antibiotic can be achieved using an implantable drug pump as a delivery system and that this method can be used to sterilize infected wounds . This method was applied in the treatment of osteomyelitis in 14 patients whose duration of symptoms ranged from one month to 22 years . In 13 patients, active drainage had been present for more than six months, and all patients had undergone one or more attempts at eradication of their infection . Prior therapy included surgical debridement alone (one patient), one or more attempts at debridement and extended intravenous antibiotic therapy (13 patients), debridement and local flap procedures (four patients), and extended oral antibiotic therapy (two patients) . The bones involved were the tibia (six patients), femur (four patients), and elbow, shoulder, hip, and radius (one patient each) . Intra-operative cultures indicated that the infecting organism was Staphylococcus aureus alone (seven patients), S . aureus in combination with gram-negative bacteria (five patients), or gram-negative organisms alone (two patients) . Debridement and pump implantation were performed in one stage . On an outpatient basis, the pumps were filled with amikacin in a concentration determined by each patient's serum level . Duration of therapy was based upon cessation of drainage and erythrocyte sedimentation rate . During therapy, serum amikacin levels ranged from less than 2.5 micrograms/ml to 8.2 micrograms/ml . The amikacin concentration in the wound drainage was always greater than the upper limits of what could be measured (greater than 55 micrograms/ml to greater than 5,000 micrograms/ml) . There were no cases of ototoxicity and one case of minimal renal toxicity (posttreatment creatinine clearance of 66 ml per minute; normal equal to 70 ml per minute) . Length of therapy ranged from 32 to 140 days (mean equal to 63 days) . Length of hospitalization ranged from five to 52 days (mean equal to 24 days) . Drainage in all patients stopped during therapy, but resumed in three patients after pump removal--in two patients, for brief periods of time, and in one patient, it continues . Twelve patients have posttreatment erythrocyte sedimentation rates less than or equal to 20 mm per hour.(ABSTRACT TRUNCATED AT 400 WORDS)

Biochemistry, 1986 Jun 17, 25(12), 3719 - 24
Biochemical characterization of phosphorylated beta-adrenergic receptors from catecholamine-desensitized turkey erythrocytes; Stadel JM et al.; Isoproterenol-induced desensitization of turkey erythrocyte adenylate cyclase is accompanied (1) by a decrease in the mobility of beta-adrenergic receptor proteins, specifically photoaffinity labeled with 125I-(p-azidobenzyl)carazolol (125I-PABC), on sodium dodecyl sulfate (SDS)-polyacrylamide gels and (2) by a 2-3-fold increase in phosphate incorporation into the beta receptor {Stadel, J.M., Nambi, P., Shorr, R . G . L., Sawyer, D . F., Caron, M . G., & Lefkowitz, R . J . (1983) Proc . Natl . Acad . Sci . U.S.A . 80, 3173} . Analysis of 32P-labeled beta receptors partially purified by affinity chromatography and subsequently hydrolyzed in 6 N HCl revealed that the beta receptor from control erythrocytes contained only phosphoserine and that agonist-promoted phosphorylation of the receptor in desensitized cells occurred on serine residues . Comparison of limited-digest peptide maps of 125I-PABC-labeled beta receptors from control and desensitized erythrocytes reveals distinctly different sensitivities of the two beta receptors to cleavage by chymotrypsin and Staphylococcus aureus protease . The altered mobility of the 125I-PABC-labeled beta receptor from desensitized erythrocytes was eliminated when 5 M urea was included in the SDS-polyacrylamide gels . Limited-digest peptide mapping of 32P-labeled beta receptors from control and desensitized cells with the protease papain identified a unique phosphorylated peptide in desensitized preparations . Our results are consistent with the hypothesis that the altered mobility of beta-receptor proteins on SDS gels following desensitization is due to changes in conformation promoted by prolonged exposure to agonists.

Eur J Biochem, 1986 Jun 16, 157(3), 497 - 506
Complete amino acid sequence of plastocyanin from a green alga, Enteromorpha prolifera; Simpson RJ et al.; The complete amino acid sequence of the plastocyanin from the green alga Enteromorpha prolifera has been determined by Edman degradation of the intact molecule and fragments produced by enzymatic cleavage of the polypeptide chain with chymotrypsin, Staphylococcus aureus protease, proline-specific endopeptidase, Lys-C endopeptidase and trypsin . The molecule consists of 98 amino acid residues with a calculated relative molecular mass of 10103 . The amino acid sequence of E . prolifera plastocyanin shows a high degree of homology with those plastocyanins from other algae and higher plants . In particular, the four residues which are copper ligands in other plastocyanins and in the bacterial electron transport protein azurin (two histidines, one cysteine and one methionine) are conserved . Five out of the six acidic amino acid side-chains which create an 'acidic patch' on the surface of plastocyanin from Populus nigra var . italica {Colman, P . M . et al . (1978) Nature (Lond.) 272, 319-324} are conserved in the amino acid sequence of E . prolifera plastocyanin.

Cancer, 1986 Jun 15, 57(12), 2343 - 5
The in vitro effects of methotrexate on the phagocytosis and intracellular killing of Staphylococcus aureus by human neutrophils; Johnson JD et al.; The in vitro effect of therapeutic concentrations of methotrexate on the phagocytosis and intracellular killing of Staphylococcus aureus by circulating human neutrophils was assessed . Neutrophils were isolated from whole blood of six healthy human volunteers by density centrifugation and incubated with 10(-3) M methotrexate . Staphylococcus aureus was opsonized in human serum and added to the prepared neutrophils . Phagocytosis was determined by serial dilutions and plating of unphagocytized bacteria . After treatment with lysostaphin and trypsin, intracellular killing of bacteria was determined at set intervals by serial dilutions and platings of viable bacteria released by neutrophil lysis . Methotrexate-treated neutrophils phagocytized 56% and control neutrophils 67% of bacteria in 15 minutes (P greater than 0.5), and 96% of ingested bacteria were killed in 15 minutes by both populations . In these experiments, the in vitro incubation of circulating human neutrophils with methotrexate produced no significant alterations in neutrophilic phagocytosis or intracellular killing of S . aureus.

J Immunol, 1986 Jun 15, 136(12), 4542 - 7
Production of B cell growth factor by normal human B cells; Jurgensen CH et al.; Although it has been demonstrated that malignant human B cell lines are capable of producing B cell growth factor (BCGF), production of BCGF by normal B cells has not been shown . In this study, we demonstrate BCGF production by normal B cells, achieved by using human peripheral blood B cells prepared by a positive selection technique and stimulated with Staphylococcus aureus Cowan I (SAC) for 12 hr . SAC was removed from the supernatants by anti-SAC-coupled Sepharose . Supernatants absorbed with this antibody were functionally free of SAC, as demonstrated by their inability to activate resting B cells . B cells stimulated with SAC for 12 hr produced BCGF activity that was generally unmeasurable in supernatants by 36 hr . Characterization of BCGF produced by SAC-stimulated B cells revealed a m.w . of 32,000 by high-performance liquid chromatography sieving and sodium dodecyl sulfate-polyacrylamide gel electrophoresis; this BCGF was found to have an isoelectric point of 6.7 . Furthermore, this BCGF lacked interleukin 1, interleukin 2, interferon, and B cell differentiation factor activity . This observation that BCGF can be produced by normal human B cells is significant because it demonstrates for the first time that normal B cells have the ability to provide their own growth factors or the growth factors for other B cells.

J Immunol, 1986 Jun 15, 136(12), 4427 - 31
The effect of 1,25-dihydroxyvitamin D3 on in vitro immunoglobulin production in human B cells; Iho S et al.; 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) dose-dependently suppressed immunoglobulin (Ig) production of human B cells, as evaluated by IgG-plaque-forming cells (IgG-PFC) in the culture of pokeweed mitogen (PWM)-activated B cells . Similar suppressive effect of 1,25(OH)2D3 on Ig production of B cells was observed in the Staphylococcus aureus Cowan I(SAC)-induced Ig-producing system . The mean percentage of inhibitions at a concentration of 10(-9) M were 60.0 +/- 8.2% (mean +/- SE, n = 6) and 65.1 +/- 4.7% (n = 10) in PWM- and SAC-stimulated cultures, respectively . The suppression was strongly exhibited only when 1,25(OH)2D3 was added at the start of the 6-day culture, accompanied by a decrease in DNA synthesis of B cells in both culture systems . On the other hand, the addition of 1,25(OH)2D3 on day 4, when DNA synthesis reached at plateau and IgG-PFC began to be detectable, had no noticeable affect on either the number of PFC or DNA synthesis of B cells . Furthermore, 1,25(OH)2D3 suppressed Ig production even when B cells were exposed to the agent for 4 hr after the activation with PWM or SAC, but not before the activation . These results indicate that 1,25(OH)2D3 inhibits B cell proliferation before differentiation to Ig-secreting cells, consequently reducing Ig production; and that its action appears to be mediated by the cytosol receptors expressed on activated B cells . Thus, the agent may serve as an immunoregulating hormone in vivo, as well as in vitro.

J Biol Chem, 1986 Jun 15, 261(17), 8028 - 35
Vascular smooth muscle caldesmon; Clark T et al.; Caldesmon, a major actin- and calmodulin-binding protein, has been identified in diverse bovine tissues, including smooth and striated muscles and various nonmuscle tissues, by denaturing polyacrylamide gel electrophoresis of tissue homogenates and immunoblotting using rabbit anti-chicken gizzard caldesmon . Caldesmon was purified from vascular smooth muscle (bovine aorta) by heat treatment of a tissue homogenate, ion-exchange chromatography, and affinity chromatography on a column of immobilized calmodulin . The isolated protein shared many properties in common with chicken gizzard caldesmon: immunological cross-reactivity, Ca2+-dependent interaction with calmodulin, Ca2+-independent interaction with F-actin, competition between actin and calmodulin for caldesmon binding only in the presence of Ca2+, and inhibition of the actin-activated Mg2+-ATPase activity of smooth muscle myosin without affecting the phosphorylation state of myosin . Maximal binding of aorta caldesmon to actin occurred at 1 mol of caldesmon: 9-10 mol of actin, and binding was unaffected by tropomyosin . Half-maximal inhibition of the actin-activated myosin Mg2+-ATPase occurred at approximately 1 mol of caldesmon: 12 mol of actin . This inhibition was also unaffected by tropomyosin . Caldesmon had no effect on the Mg2+-ATPase activity of smooth muscle myosin in the absence of actin . Bovine aorta and chicken gizzard caldesmons differed in several respects: Mr (149,000 for bovine aorta caldesmon and 141,000 for chicken gizzard caldesmon), extinction coefficient (E1%280nm = 19.5 and 5.0 for bovine aorta and chicken gizzard caldesmon, respectively), amino acid composition, and one-dimensional peptide maps obtained by limited chymotryptic and Staphylococcus aureus V8 protease digestion . In a competitive enzyme-linked immunosorbent assay, using anti-chicken gizzard caldesmon, a 174-fold molar excess of bovine aorta caldesmon relative to chicken gizzard caldesmon was required for half-maximal inhibition . These studies establish the widespread tissue and species distribution of caldesmon and indicate that vascular smooth muscle caldesmon exhibits physicochemical differences yet structural and functional similarities to caldesmon isolated from chicken gizzard.

J Immunol Methods, 1986 Jun 10, 90(1), 111 - 23
Establishment of a human B cell line that proliferates in response to B cell growth factor; Abe H et al.; A human B cell line which shows a marked dose dependence on B cell growth factor (BCGF) when cultured in less than or equal to 2% serum has been established . Human B lymphocytes were obtained from peripheral blood of normal donors and cultured in the presence of anti-IgM (mu chain specific) and BCGF . Frequent refeedings with fresh medium containing BCGF and anti-IgM led to the establishment of a long term cultured human B cell line, HAB-40 . Phenotyping of HAB-40 revealed that the cell population consisted predominantly of IgM-bearing (72%) and B1 (100%) positive cells . This B cell line consistently secreted IgM and IgG when co-cultured in the presence of PMA, anti-IgM and beta or gamma interferon (IFN) . Also, it was Epstein-Barr virus nuclear antigen (EBNA) positive (100%) . HAB-40 cells have been successfully maintained in the presence of BCGF without anti-IgM for over a year . Removal of BCGF led to the rapid loss of viable cells in cultures containing less than 2% serum . HAB-40 cells in microassays exhibited a marked dose-dependent incorporation of {3H}thymidine in response to BCGF in the absence of any exogenous stimulants such as anti-IgM or Staphylococcus aureus Cowan I (SAC) . Recombinant interleukin 2 (IL-2) failed to augment the {3H}thymidine uptake by these B cells despite the low density expression of Tac antigen (IL-2 receptor) on their cell surface, or even when the cells were stimulated with phorbol myristate acetate (PMA) to express higher density of Tac antigen (48%) . HAB-40 cells could be maintained in BCGF which was partially purified to deplete it of other contaminating proteins . None of the seven well established EBNA-positive human B cell lines nor two chronic B lymphocytic leukemia (B-CLL) cell lines that were tested showed BCGF dependence . The same BCGF-active chromatographic fractions that were active on HAB-40 cells also stimulated BCL1 and normal human B cells stimulated with anti-IgM . In the presence of less than or equal to 2% serum proteins this cell line provides a simple, reproducible assay for BCGF even in the presence of contaminant IL-2.

Biochim Biophys Acta, 1986 Jun 5, 871(2), 217 - 23
Primary structure of the cholesterol side-chain cleavage cytochrome P-450 from bovine adrenocortical mitochondria and some aspects of its functioning on a structural level; Chashchin VL et al.; The primary structure of the cholesterol side-chain cleavage cytochrome P-450 (P-450scc) from bovine adrenocortical mitochondria has been determined . At the initial stage an exhaustive chymotryptic digestion of carboxymethylated P-450scc was performed, and the amino acid sequence of 66 peptides was determined . At the second stage an investigation of the amino acid sequence of individual fragments I (Mr 29 800) and II (Mr 26 600) of the limited trypsinolysis of P-450scc was carried out . Fragment I was digested with trypsin, Staphylococcus aureus V8 proteinase and thermolysin; fragment II was cleaved with trypsin and S . aureus V8 proteinase . In addition, the amino acid sequence of some CNBr peptides of P-450scc has been investigated . The primary structure of cytochrome P-450scc determined with protein chemistry methods proved the multistage cholesterol transformation to pregnenolone to be catalyzed by a single species of cytochrome P-450scc which consists of 481 amino acids . The results from protein sequencing of P-450scc are in good agreement with those obtained recently from nucleotide sequencing . The localization of peptide bonds cleaved under limited proteolysis of P-450 with trypsin to fragments I and II, I and III (Mr 16 800) is presented . It is shown that the transformation of P-450scc to P-420 is accompanied by the appearance of an additional trypsin-sensitive peptide bond in the N-terminal part of P-450scc.

Biochim Biophys Acta, 1986 Jun 5, 871(2), 177 - 81
Effect of calcium binding on conformational changes of staphylococcal metalloproteinase measured by means of intrinsic protein fluorescence; Wasylewski Z et al.; The removal by EDTA of Ca2+ from the two-tryptophan-containing metalloproteinase isolated from Staphylococcus aureus leads to an increase in its intrinsic fluorescence intensity . Based on acrylamide fluorescence quenching results, analyzed by the non-linear least-squares method, we have shown that this protein molecule undergoes irreversible conformational change upon removal of Ca2+, which include the exposure to the solvent of buried tryptophan residues . Steady-state fluorescence anisotropy measurements indicate that the loss of Ca2+ leads to a significant increase in internal mobility of previously buried tryptophan residues.

J Biol Chem, 1986 Jun 5, 261(16), 7236 - 41
Soluble guanylate cyclase from rat lung exists as a heterodimer; Kamisaki Y et al.; The soluble form of guanylate cyclase (EC 4.6.1.2) from rat lung has been purified to homogeneity by a one-step immunoaffinity chromatographic procedure . The purified soluble guanylate cyclase has specific activities of 432 and 49.1 nmol of cyclic GMP formed per min/mg protein with manganese and magnesium ions as a cofactor, respectively . This represents a purification of approximately 2,000-fold with a 50% recovery . The native enzyme has a molecular weight of 150,000 and a Stokes radius of 4.8 nm as determined on Spherogel TSK-G3000SW gel permeation chromatography . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis results in two protein-staining bands with molecular weights of 82,000 and 70,000 . The purified soluble guanylate cyclase was also subjected to native polyacrylamide gel electrophoresis, isoelectric focusing electrophoresis, ion exchange chromatography, and GTP-agarose affinity chromatography . These additional purification procedures confirmed the presence of a single protein peak coincident with enzyme activity . The two subunits separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis were shown to have different primary structures by immunoblotting with monoclonal and polyclonal antibodies prepared against purified soluble guanylate cyclase and by peptide mapping with papain or Staphylococcus aureus V8 protease treatment . These data demonstrate that soluble guanylate cyclase purified from rat lung is a heterodimer composed of 82,000- and 70,000-dalton subunits with different primary structures.

Biochemistry, 1986 Jun 3, 25(11), 3156 - 63
Limited proteolysis of human von Willebrand factor by Staphylococcus aureus V-8 protease: isolation and partial characterization of a platelet-binding domain; Girma JP et al.; Purified human von Willebrand factor (vWF) was digested with Staphylococcus aureus V-8 protease, and specific domains interacting with platelets were isolated and characterized . Amino acid sequence analysis and sodium dodecyl sulfate gel electrophoresis demonstrated that the digestion proceeded primarily by a single cleavage of the native 270K subunit between an internal Glu-Glu peptide bond . This produced an integral stepwise degradation of the multimers of vWF with a concomitant accumulation of bands with mobility similar to that of the smaller molecular weight vWF multimers . The immediate precursor of the final products contained equimolar amounts of 270K subunit and of two polypeptides (170K and 110K) . The cleavage of the remaining 270K subunit converted vWF into two main fragments (fragments II and III) . These fragments were isolated by ion exchange chromatography, characterized, and assayed for platelet binding in the presence of ristocetin . Fragment III is a dimer of 315K composed primarily of two chains of 170K . Amino acid sequence analysis indicated that it originated from the amino-terminal portion of the 270K subunit and contained 11% of the original ristocetin cofactor activity . Also, it binds to platelets at the same specific sites as native vWF and shows a platelet binding pattern similar to that of partially reduced vWF (500K) . Fragment II is a dimer of 235K composed of two identical chains of 110K . Amino acid sequence analysis indicated that it originated from the carboxyl-terminal portion of the 270K subunit and lacked ristocetin cofactor activity . Also, it does not bind to platelets or inhibit the binding of 125I-vWF in the presence of ristocetin.(ABSTRACT TRUNCATED AT 250 WORDS)

Biken J, 1986 Jun, 29(2), 39 - 44
Effect of the composition of reversion medium on change of Staphylococcus aureus lysostaphin protoplasts to coccal forms and L-forms; Kato Y et al.; The experimental conditions under which protoplasts of Staphylococcus aureus strain MS353 (pCp) are converted to the coccal or L-form were investigated . Protoplasts prepared by treating coccal MS353 (pCp) strain with Lysostaphin formed various types of colonies (coccal form, L-form and mixed types) in about 50% yield when they were plated on reversion (R) medium consisting of 2% brain heart infusion, 0.5M sodium succinate, 0.01% bovine serum albumin, 20 mM MgCl2 and 0.6% agar . The L-form type colonies with a typical fried-egg appearance that developed on the R medium at an early stage gradually reverted to the coccal form through a mixed type stage in which a high density area first appeared in the periphery of the colony and then spread throughout the colony . The use of modified R medium without MgCl2 or R medium in which 0.5M sodium succinate as an osmotic stabilizer was replaced by 7.5% NaCl resulted in marked delay in the appearance of reverted cells . R medium without bovine serum albumin yielded atypical L-form type colonies, which contained masses of coccal cells with very irregular margins . On the other hand, R medium without MgCl2 but with penicillin G supported development of L-form type colonies at high rate (13-15%) from the inoculated protoplasts.

Antimicrob Agents Chemother, 1986 Jun, 29(6), 1032 - 9
Influence of human monocytes on the antibacterial activity of kanamycin and gentamicin for Staphylococcus aureus; van den Broek PJ et al.; The present study was performed to compare the antibacterial activities of kanamycin and gentamicin on Staphylococcus aureus phagocytosed by human monocytes and on nonphagocytosed S . aureus . The method used permitted the measurement of the effect of antibiotics on intracellular bacteria independent of phagocytosis and intracellular killing by the monocytes . A morphological assay with lysostaphin established the intracellular localization of about 70% of the cell-associated S . aureus in the monocyte-bacterium suspension . After 1 h of incubation, the antibacterial activity of both aminoglycosides was greater against intracellular than against nonphagocytosed S . aureus, but after 3 h, the reverse was true . The maximal effect on phagocytosed S . aureus, i.e., killing of about 98% of the bacteria, was reached in the first hour of incubation at kanamycin and gentamicin concentrations of 5 and 1 microgram/ml, respectively . A cell-free medium in which monocytes had been incubated increased the antibacterial activity of kanamycin, indicating that monocytes secrete a factor that enhances the antibacterial activity of aminoglycosides.

Am J Med, 1986 Jun, 80(6), 1208 - 12
Development of antibiotic resistance by staphylococcus aureus in a single patient . Confirmation by phage typing, antibiograms, and plasmid analysis; Kozarsky PE et al.; A patient is described who had colonization and infection with strains of Staphylococcus aureus increasingly resistant to multiple antibiotics while receiving several courses of broad-spectrum antibiotics . S . aureus strains with different phage types, antibiograms, and plasmid profiles were isolated from different sites at the same time . After the patient was transferred to an intermediate-care service where he received only supportive care and no further antibiotics, culture samples from areas previously colonized or infected (sputum, anterior nares, perineal region, rectum) showed no S . aureus . This patient graphically demonstrates that antibiotics may create an environment allowing the overgrowth of resistant organisms and that withdrawal of this selection pressure may allow more sensitive organisms to recolonize.

J Trauma, 1986 Jun, 26(6), 534 - 7
Neutrophil subpopulations change after thermal injury; Deitch EA et al.; Just as there are subpopulations of mononuclear leukocytes which are functionally distinct, so there appear to be different subpopulations of neutrophils, with different functional abilities . Neutrophils which display the Fc receptor (Fc+) for immunoglobulin have increased chemotactic phagocytic and bactericidal activity compared with neutrophils which are Fc receptor negative (Fc-) . To determine if the acquired neutrophil dysfunction which occurs after thermal injury could be due to a change in the percentage of Fc+ neutrophils, serial studies of neutrophil function, including random migration, chemotaxis, phagocytosis, and killing of Staphylococcus aureus, were performed in 12 patients and related to the percentage of neutrophils which possessed the Fc receptor . After thermal injury the percentage of Fc+ cells decreased significantly (p less than 0.01) . However, no correlation between the number of rosette-forming cells and random migration (p = 0.48), chemotaxis (p = 0.45), or bactericidal activity (p = 0.50) was found in this patient population . Thus, although thermal injury was associated with a significant decrease in the number of Fc+ neutrophils, this change in neutrophil subpopulation levels did not explain the acquired defect in neutrophil function which occurred after thermal injury.

J Med Microbiol, 1986 Jun, 21(4), 343 - 7
Staphylococcus aureus protein A--antibody-mediated haemagglutination for the analysis of cross-reactivity between phenol-water extracts of Bacteroides fragilis; Srinivasakumar N et al.; Passive haemagglutination (PHA) and Staphylococcus aureus protein A--antibody-mediated haemagglutination (SAPA-AMHA) were used to analyse the cross-reactions of rabbit antisera against four strains of Bacteroides fragilis . There was cross-reactivity between all the strains tested but strain-specific reactions were obtained with three strains . Two to 32-fold higher antibody titres were obtained with SAPA-AMHA than with PHA . The antigen concentration required in inhibition assays was up to 32-fold higher for PHA than for SAPA-AMHA . The latter is a simple and superior alternative to PHA for such studies.

Arch Intern Med, 1986 Jun, 146(6), 1118 - 21
Staphylococcus aureus endocarditis . A review of 119 cases; Espersen F et al.; Staphylococcus aureus endocarditis cases in Denmark from 1976 to 1981 were reviewed . A total of 119 patients--61 female and 58 male, with a median age of 63 years (range, 1 month to 85 years)--fulfilled the diagnostic criteria . Community-acquired infections were most common (62%), but the frequency of hospital-acquired cases (38%) was greater than in earlier reports . The clinical picture was relatively nonspecific, and 32% of the patients had no heart murmurs initially . In 65 cases (55%), endocarditis was not suspected clinically, and the diagnosis was first obtained at autopsy . The mortality was 71% and correlated with age, hospital-acquired infection, and the presence of heart failure and arterial embolism.

South Med J, 1986 Jun, 79(6), 696 - 7
Chronic mandibular osteomyelitis; Harris LF; Chronic osteomyelitis of the mandible is an infrequently reported condition, but recent experience with six cases over a 14-month period suggests it is more common than appreciated . Chronic mandibular osteomyelitis results from odontogenic infection, postextraction complication, trauma, or irradiation to the mandible . Clinical findings include local pain and swelling and trismus, but constitutional symptoms are unusual . Radiologic examination discloses radiolucent areas, bony destruction, and sequestrum formation . Pathogenic organisms are normal oral flora, Staphylococcus aureus, and aerobic gram-negative bacilli . Chronic mandibular osteomyelitis must be differentiated from malignant disease involving the mandible . Diagnosis is accomplished by bone biopsy and culture . Treatment involves through surgical debridement and prolonged antimicrobial therapy . Osteoradionecrosis of the mandible is extremely recalcitrant to conventional therapy, but aggressive surgery and treatment have proven effective.

Mol Pharmacol, 1986 Jun, 29(6), 649 - 56
Location of a polypeptide sequence within the alpha-subunit of the acetylcholine receptor containing the cholinergic binding site; Oblas B et al.; Proteolytic fragments of the alpha-subunit of the acetylcholine receptor of Torpedo electric organ were generated by digestion with Staphylococcus aureus V8 protease, and their ability to bind alpha-bungarotoxin was assessed following resolution on polyacrylamide gels and transfer to nitrocellulose . The position of the smallest fragment (Mr = 17,000) with toxin-binding activity was located within the primary sequence of the alpha-subunit by isolation and chemical characterization . The amino acid sequence at its amino terminus is Val-Asn-Gln-Ile-Val-Glu, which is identical to a unique sequence on the alpha-subunit beginning at Val 46 . Based on considerations of the apparent molecular weight of this polypeptide fragment, the enzyme specificity of V8 protease, analysis of the partial amino acid composition, and the position of various identifying landmarks of the primary sequence of the alpha-subunit, the carboxy terminus is restricted to an acidic amino acid residue between Asp 152 and Asp 180 . The apparent affinities of the 17-kDa polypeptide for alpha-bungarotoxin (IC50 = 100 nM) and d-tubocurarine (IC50 = 0.4 mM) were not significantly different from the values obtained for the undigested alpha-subunit, suggesting that the 17-kDa polypeptide contains all of the essential elements present on the alpha-subunit that are necessary for the binding of both of these cholinergic ligands . A second toxin-binding fragment (Mr = 19,000) was also identified . Although the amino terminus of this polypeptide fragment has not been determined directly, its position relative to Asn 141, the site of N-glycosylation, was established, and it appears that its amino terminus cannot be located any closer to the amino terminus of the alpha-subunit than Asp 152 . If this tentative assignment is correct, then the region of the alpha-subunit involved in binding cholinergic ligands is more closely defined as occurring within the segment of the primary sequence of the alpha-subunit that the 17- and 19-kDa polypeptide fragments have in common, i.e., an overlapping sequence maximally bounded by Asp 152 and Asp 180.

J Clin Microbiol, 1986 Jun, 23(6), 1030 - 3
Colonization of newly arrived house staff by virulent staphylococcal phage types endemic to a hospital environment; Ballou WR et al.; The acquisition of hospital strains of Staphylococcus aureus by new house officers was studied in an 800-bed referral hospital over a 1-year period . S . aureus isolates, including three strains with characteristic phage patterns that had previously been documented to cause disease in patients and colonize hospital personnel, were recovered from the anterior nares of 35 of 54 newly arrived house officers . There was a significant correlation (r = 0.7475; P less than 0.02) between colonization with the dominant hospital strain (S) and exposure to the hospital environment over 12 months . No hospital-wide increase in infections owing to the S strain was seen during this period, which suggests that house staff acquired this strain from reservoirs within the hospital . The finding of colonization with virulent endemic S . aureus strains in house officers working on every ward of the hospital suggests that new strategies for control of S . aureus nosocomial infections must be considered and evaluated.

Infect Immun, 1986 Jun, 52(3), 714 - 7
Binding of Staphylococcus aureus by human serum spreading factor in an in vitro assay; Fuquay JI et al.; Human serum spreading factor, an animal cell adhesion-promoting glycoprotein, bound Staphylococcus aureus in an assay in which association of bacteria to protein-precoated microtiter wells was quantitated . Interaction of human serum spreading factor with S . aureus suggests that this protein may play a role in host defenses and in the invasiveness and pathogenicity of microbial infections.

Infect Immun, 1986 Jun, 52(3), 671 - 5
Detection of staphylococcal membrane receptors on virus-infected cells by direct adhesin overlay; Sanford BA et al.; We partially characterized the interaction between 125I-labeled surface proteins of Staphylococcus aureus, obtained by thermal extraction, and purified plasma membranes from uninfected and influenza virus A/FM/1/47-infected Madin-Darby canine kidney cells . The radioactivity profile of surface-labeled proteins, derived from anion-exchange high-pressure liquid chromatography, showed a mixture of acidic polypeptides in a peak which contained approximately 5% of the total protein injected; adsorption with purified plasma membranes reduced radioactivity by 5 to 7% and altered elution profiles . Using a direct adhesin overlay procedure, we found that these surface-labeled proteins reacted with polypeptides located on the external surface of plasma membranes which were shared by both virus-infected and uninfected cells or were unique to virus-infected cells . Our data may help explain the enhanced adherence of S . aureus to influenza virus A-infected cells in vitro.

J Gen Microbiol, 1986 Jun, 132 ( Pt 6), 1723 - 8
Detection of an integrated tetracycline resistance plasmid in the chromosome of methicillin-resistant Staphylococcus aureus; Gillespie MT et al.; The majority of multiresistant Staphylococcus aureus strains isolated in Australian hospitals since 1970 carry a chromosomally-encoded minocycline and tetracycline resistance determinant . By using DNA-DNA hybridization, some of these multiresistant strains were shown to possess also a tetracycline resistance plasmid, equivalent to pT181, integrated into a unique site in the chromosome . By relating the hybridization data to the map of pT181, the site of integration on this plasmid was established to be between the genes for replication and tetracycline resistance.

J Hyg (Lond), 1986 Jun, 96(3), 419 - 23
In vitro and in vivo study of fosfomycin in methicillin-resistant Staphylococcus aureus septicaemia; Lau WY et al.; Five hundred strains of methicillin-resistant Staphylococcus aureus were tested against various anti-staphylococcal agents . Vancomycin, fusidic acid and fosfomycin were found to be the most effective . Only 1 strain out of 500 was resistant to fosfomycin . Three patients with methicillin-resistant Staphylococcus aureus septicaemia were successfully treated by fosfomycin . We conclude that fosfomycin could be the drug of choice for methicillin-resistant Staphylococcus aureus infection.

J Antimicrob Chemother, 1986 Jun, 17(6), 705 - 15
Physiological determination of methicillin resistance in Staphylococcus aureus: comparison of clinical and genetically derived isolates; Heneine N et al.; The expression of methicillin resistance in 10 clinical and 5 genetically constructed strains of Staphylococcus aureus has been measured in relation to temperature of incubation (37 degrees C versus 30 degrees C), the presence of additional sodium chloride in the medium (5.5% vs . 0.5%), and pH (7.4 vs . 5.4) . Resistance was quantitatively measured by disc diffusion and agar dilution assays . In disc assays of methicillin resistance, all but one clinical isolate and both transduced strains displayed increased the resistance at the lower temperature . Increased salt had little effect, or decreased the resistance of these strains . At pH 5.4 resistance decreased substantially . Methicillin resistance in three step-selected strains, by contrast, was variably affected by changes in temperature or salt concentration, though the effects were never great . The most distinctive feature of these strains was that pH change did not affect their resistance . In agar dilution assays, where surviving fractions within the population were measured as a function of antibiotic concentration, the temperature and salt effects seen in disc assays of resistance were again evident for the two clinical isolates tested, but not for the transduced strains, where added salt increased resistance at the higher concentrations of methicillin tested . For two step-selected strains, the effects of lowering temperature and increasing salt on resistance were variable . By contrast with the disc assays, these strains were less resistant at the lower pH, though not to the same relative extent as that shown by the clinical isolates and transduced strains . Resistance heterogeneity was not significantly affected by higher salt and lower incubation temperature in the two clinical strains tested, but elevated salt consistently increased heterogeneity in transduced and step-selected strains at the higher concentrations of methicillin tested . In the absence of salt, step-selected strains exhibited little heterogeneity, unlike the clinical and transduced strains . We conclude that the similarities and differences seen in clinical isolates and laboratory strains favour the idea that methicillin resistance arises clinically in S . aureus as a result of transduction (or other genetic transfer) of existing gene(s) determining resistance, rather than as a result of selection of mutants among sensitive or less resistant strains . Our findings also indicate that inclusion of salt in media containing methicillin may attenuate resistance to the antibiotic and thus decrease the sensitivity of detection of marginally resistant strains of S . aureus.

Infect Control, 1986 Jun, 7(6), 317 - 20
Strict versus modified isolation for prevention of nosocomial transmission of methicillin-resistant Staphylococcus aureus; Ribner BS et al.; Patients colonized or infected with methicillin-resistant Staphylococcus aureus (MRSA) in a Surgical Intensive Care Unit and Surgical Intermediate Care Unit were placed either in Strict Isolation or cared for with modified isolation precautions . The assignment was determined by the unit in which they were hospitalized . Units were changed from one form of isolation to the other and served as their own controls . Over a 4-month study period, the rate of MRSA transmission did not change when the type of isolation precautions were altered . The ratio of colonized to infected patients also remained constant . Infected patients were usually first detected by clinical specimens, while colonized patients were usually detected by surveillance cultures performed under the study protocol . Following the study, all hospitalized patients with MRSA were placed in modified isolation precautions . Total new acquisitions of MRSA in the hospital have decreased over the subsequent 6-month period.

Acta Pathol Microbiol Immunol Scand {C}, 1986 Jun, 94(3), 91 - 6
The mitogenic properties of lipoteichoic acid from Staphylococcus aureus; Aasjord P et al.; Lipoteichoic acid (LTA) isolated from Staphylococcus aureus Cowan I induced an in vitro blastogenic response in isolated mononuclear cells (MNC) . Seventeen of 20 normal donors responded strongly, with peak response after 3-5 days of incubation . Significant stimulation was also obtained with human cord blood MNC from 3 donors, further indicating that the stimulation was due to a general mitogenic effect . The mitogenicity of deacylated LTA (dLTA) was similar to that of LTA . Binding of LTA to isolated MNC was demonstrated by an indirect immunofluorescent technique staining with specific rabbit anti-LTA antibodies and by rosette inhibition experiments . Results obtained in cell fraction experiments indicated that LTA had a monocyte dependent mitogenic effect on T lymphocytes.

J Appl Bacteriol, 1986 Jun, 60(6), 535 - 44
The adherence of Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli on cotton, polyester and their blends; Hsieh YL et al.; The adherent behaviour of the Gram-positive Staphylococcus aureus and Staphylococcus epidermidis and the Gram-negative Escherichia coli on cotton, polyester and their blends through contact in aqueous suspensions was studied . Staphylococcus epidermidis was found to adhere to fabrics much more so than Staph . aureus . The adherence of both Staph . epidermidis and Staph . aureus to fabrics increased as the content of polyester fibres in the fabrics increased . The attachment of E . coli to all fabrics was very low and was not affected by the fibre contents . Total numbers of adherent bacteria on cotton and polyester fabrics were related directly to the concentrations of the bacterial suspensions . The extents of adherence, expressed by the percentage of adherent bacteria from the suspension, however, were independent of the concentration . The length of contact with bacteria was also found to affect the adherence of bacteria on fabrics studied.

Biokhimiia, 1986 Jun, 51(6), 909 - 15
{Various physico-chemical properties of lytic proteinase L2 of the enzyme preparation lysoamidase isolated from bacteria of Pseudomonadaceae family}; Stepnaia OA et al.; Some physico-chemical properties of lytic proteinase L2 isolated from the enzymatic microbial preparation of lysoamidase were studied . The molecular mass of the enzyme is 15 000 Da, pI is 5.3 . The enzyme hydrolyzes casein as well as the cells and cell walls of Staphylococcus aureus 209-P . The pH optimum of casein hydrolysis lies at 9.5; that for cell wall hydrolysis at 8.0 . The temperature optimum for casein hydrolysis and cell lysis lies at 55 degrees C and 65 degrees C, respectively . The enzyme proteolytic activity is inhibited by serine proteinase inhibitors in a greater degree than the lytic activity . 50% of the proteolytic and lytic activities is lost upon enzyme heating for 15 min at 65 degrees C.

Antimicrob Agents Chemother, 1986 Jun, 29(6), 965 - 8
Canine model for the simultaneous measurement of antibiotic levels in tissues and bacterial killing rate; Wagner DS et al.; Antibiotic levels in serum are commonly used to guide antibiotic therapy . The antibiotic levels in interstitial fluid are a more accurate reflection of the efficacy of antibiotic penetration into the tissues . Although there are experimental models for determining interstitial fluid levels, there is no model for measuring the in vivo killing of bacteria, which is the endpoint of antibiotic therapy . We developed an accurate, reliable animal model which allows measurement of the in vivo killing of bacteria along with a determination of antibiotic levels in tissues . Modified Sykes-Moore chambers were applied to the dissected external oblique muscle of 14 dogs . The chambers were inoculated with clinical isolates of Staphylococcus aureus or Escherichia coli . The dogs were treated with cefoxitin or gentamicin . Quantitative cultures were performed, and the antibiotic levels in interstitial fluid were determined.

Antimicrob Agents Chemother, 1986 Jun, 29(6), 1092 - 4
In vitro and in vivo efficacy of the combination trimethoprim-sulfamethoxazole against clinical isolates of methicillin-resistant Staphylococcus aureus; Elwell LP et al.; The in vitro susceptibilities of 16 independent, geographically distinct clinical isolates of methicillin-resistant Staphylococcus aureus to trimethoprim (TMP) in combination with sulfamethoxazole (SMX) were evaluated . Although methicillin-resistant S . aureus strains appear to be universally resistant to SMX, the combination TMP-SMX was found to be synergistic in vitro (in combination, the MICs of both drugs decreased 6- to 25-fold) as well as in vivo (5- to 6-fold reduction in TMP at 50% effective doses).

J Clin Microbiol, 1986 Jun, 23(6), 997 - 1000
Influence of technical factor variations on serum inhibition and bactericidal titers; Woolfrey BF et al.; The influence of technical factor variations on serum bactericidal and serum inhibitory titers was studied by using Staphylococcus aureus clinical isolates versus oxacillin-spiked human serum . Parallel tests, both with and without the use of beta-lactamase in count plates to inactivate oxacillin carryover, were performed with a conventional macrodilution approach, a carefully controlled macrodilution procedure, and a standard microdilution method . Careful control of technical factor variations diminished the incidence of low serum bactericidal titers and decreased the dispersion of results, a finding corollary to the known influence of technical factor variations on the measurement of MBCs . The incorporation of beta-lactamase into count plates resulted in a shift of serum bactericidal titers to lower values . The microdilution method appeared to be least influenced by technical variations and, with the addition of beta-lactamase to count plates, provided the best results.

Immunology, 1986 Jun, 58(2), 203 - 8
Production of human and murine interleukin-2 by toxic shock syndrome toxin-1; Micusan VV et al.; Toxic shock syndrome toxin-1 (TSST-1), isolated from Staphylococcus aureus strains associated with toxic shock syndrome (TSS), is known as a potent mitogen and interleukin-1 inducer . The potential of TSST-1 as an interleukin-2 (IL-2) inducer was tested on human peripheral blood lymphocytes (HPBL) and murine spleen lymphocytes (MSL) . These cells were incubated with TSST-1 and the supernatants analysed for IL-2 production . Preincubation of IL-2-dependent indicator cells (IC) with a monoclonal antibody specific for murine IL-2 receptors inhibited their proliferation by supernatants of TSST-1-treated MSL, thus strongly suggesting that they contain IL-2 . The concentrations of TSST-1 required for HPBL or MSL to produce IL-2 ranged between 10(-1) and 10(-4) micrograms/ml . The amount of IL-2 units/ml varied little from one experiment to another . In contrast, IL-2 production by PHA-stimulated HPBL or Con A-stimulated MSL showed great variability and dependence on mitogen concentration . T-cell depleted MSL exposed to TSST-1 produced less IL-2 . Experiments with germ-free mice and TSST-1-primed mice demonstrated that IL-2 production is not related to TSST-1 antigenicity.

Orthop Rev, 1986 Jun, 15(6), 387 - 92
Atraumatic removal of a well-fixed porous ingrowth hip prosthesis; McClelland SJ et al.; The authors describe their experience in the removal of a firmly fixed porous coated femoral component of a bipolar hip prosthesis in a 36-year-old man on a methadone maintenance program . The uncemented prosthesis had been implanted for 15 months when the patient complained of hip pain . A methicillin-resistant Staphylococcus aureus infection was found in the hip joint . It was decided to remove the femoral component when the infection failed to respond to irrigation, debridement, and several weeks of intravenous antibiotic therapy and because the patient developed early signs of impending renal compromise and a deterioration of his general nutritional status . The technique for separating and removing the well-fixed porous ingrowth component from its intramedullary environment without damaging the cortical tube of the proximal femur is described.

Acta Pathol Microbiol Immunol Scand {B}, 1986 Jun, 94(3), 127 - 33
Antigenic and biochemical characteristics of Mobiluncus mulieris and Mobiluncus curtisii; Skarin A; Twenty-four strains of Mobiluncus mulieris and 27 strains of Mobiluncus curtisii were tested with respect to 6 different biochemical characteristics: arginin-decarboxylase activity, beta-galactosidase activity, synergistic hemolysis with Staphylococcus aureus, hydrolysis of hippurate, migration through soft agar and reduction of nitrate . Antigens of the same strains, prepared by ultrasonication, were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis followed by immunoblotting using polyclonal rabbit antisera against two of the M . mulieris strains and five of the M . curtisii strains . Two different strongly reacting protein antigens could be detected in the M . mulieris strains . These strains could be separated into three groups based on the possession of either of the two antigens or both . In the M . curtisii strains, 10 strongly reacting protein antigens could be detected . Four strains possessed only one of these antigens, one did not possess any, while the remaining strains possessed different sets of 9 of them . Within each species common protein antigens were detected . No antigens were found which were shared by both species . The biochemical characteristics studied could not differentiate between the antigenic groups in any of the species . None of the antigenic subgroups of M . curtisii found in the present study was identical with any of the two subspecies (curtisii and holmesii) which have been proposed.

J Antimicrob Chemother, 1986 Jun, 17(6), 767 - 74
Comparison of the effect of phenoxymethylpenicillin, cloxacillin, and flucloxacillin on Staphylococcus aureus phagocytosed by human monocytes; van den Broek PJ et al.; The antibacterial effect of phenoxymethylpenicillin, cloxacillin, and flucloxacillin on pre-opsonized Staphylococcus aureus ingested by human monocytes and on preopsonized S . aureus in suspension was compared . The antibiotics were 1.7-3 times more effective against intracellular S . aureus than against S . aureus in suspension . No influence of acid stability or lipid solubility of the drugs on the antibacterial effect against intracellular S . aureus was demonstrated.

Anal Biochem, 1986 Jun, 155(2), 395 - 9
A radioimmune assay of ganglioside GM1 synthase using cholera toxin; Honke K et al.; A radioimmune assay for uridine 5'-diphosphate-galactose (UDP-Gal):GM2 galactosyltransferase, which synthesizes GM1, has been developed utilizing cholera toxin . This assay is more sensitive and simpler than previously used assays . Radioactive nucleotide substrate and GM2 were incubated with an enzyme sample, and a radiolabeled product, GM1, was reacted with cholera toxin . The GM1-cholera toxin complex was further reacted with anti-cholera toxin and Staphylococcus aureus cell suspension . The resulting complex was transferred onto a nitrocellulose membrane and quantitated by liquid scintillation counting . This assay was found to be sensitive for the detection of 100 pmol of the reaction product, GM1 . With this assay method, some properties of the crude enzyme extracts from rat liver were studied . The enzyme had a pH optimum of 6.5-7.0 and required Mn2+ . The Km values for UDP-Gal and GM2 were 0.12 mM and 6 microM, respectively.

J Immunol, 1986 Jun 1, 136(11), 4019 - 26
Expression and function of an early activation marker restricted to human B cells; Kikutani H et al.; A B cell-specific monoclonal antibody (anti-Ba) was prepared . In two-color FACS analysis the anti-Ba reacted with a subpopulation of Ig+ or B1+ cells obtained from tonsils, but did not react with most B1+ cells derived from PBL . Activation of B cells from PBL with TPA or anti-mu induced Ba expression and the addition of PHA-conditioned supernatant with anti-mu-enhanced Ba expression . Other B cell activators, such as Staphylococcus aureus Cowan I (Staph-A) or PWM plus T cells, could induce Ba expression . Ba expression was observed 6 hr after stimulation and reached a peak level at 72 hr . Ba expression was strictly restricted to B cells . H-7, a specific inhibitor of protein kinase C (C-kinase), displayed a dose-dependent inhibitory effect on Ba expression, showing dependency on C-kinase for Ba expression . Anti-Ba inhibited B cell proliferation induced by anti-mu and B-BCGF distinct from BSF-1 . The results presented in this study suggest that the Ba antigen on B cells may be comparable to the Tac antigen on T cells.

J Cell Biol, 1986 Jun, 102(6), 2211 - 22
Early events elicited by bombesin and structurally related peptides in quiescent Swiss 3T3 cells . I . Activation of protein kinase C and inhibition of epidermal growth factor binding; Zachary I et al.; Addition of bombesin to quiescent cultures of Swiss 3T3 cells caused a rapid increase in the phosphorylation of an Mr 80,000 cellular protein (designated 80k) . The effect was both concentration and time dependent; enhancement in 80k phosphorylation could be detected as early as 10 s after the addition of peptide . Recently, a rapid increase in the phosphorylation of an 80k cellular protein after treatment with phorbol esters or diacylglycerol has been shown to reflect the activation of protein kinase C in intact fibroblasts (Rozengurt, E., A . Rodriguez-Pena, and K . A . Smith, 1983, Proc . Natl . Acad . Sci . USA., 80:7244-7248; Rozengurt, E., A . Rodriguez-Pena, M . Coombs, and J . Sinnett-Smith, 1984, Proc . Natl . Acad . Sci . USA., 81:5748-5752) . The 80k phosphoproteins generated in response to bombesin and to phorbol 12,13-dibutyrate were identical as judged by one- and two-dimensional PAGE and by peptide mapping after partial proteolysis with Staphylococcus aureus V8 protease . In addition, prolonged pretreatment of 3T3 cells with phorbol 12,13-dibutyrate, which leads to the disappearance of protein kinase C activity, blocked the ability of bombesin to stimulate 80k . Bombesin also caused a rapid (1 min) inhibition of 125I-labeled epidermal growth factor (125I-EGF) binding to Swiss 3T3 cells . The inhibition was both concentration and temperature dependent and resulted from a marked decrease in the affinity of the EGF receptor for its ligand . Peptides structurally related to bombesin, including gastrin-releasing peptide, also stimulated 80k phosphorylation and inhibited 125I-EGF binding; both effects were selectively blocked by a novel bombesin antagonist . These results strongly suggest that these responses are mediated by specific high-affinity receptors that recognize the peptides of the bombesin family in Swiss 3T3 cells . While an increase in cytosolic Ca2+ concentration does not mediate the bombesin inhibition of 125I-EGF binding, the activation of protein kinase C in intact Swiss 3T3 cells by peptides of the bombesin family may lead to rapid inhibition of the binding of 125I-EGF to its cellular receptor.

Biken J, 1986 Jun, 29(2), 45 - 9
Prophage type of recombinants produced by cell fusion between various combinations of lysogenic or non-lysogenic substrains from two Staphylococcus aureus L-forms; Hirachi Y et al.; Streptomycin (SM)- or erythromycin (EM)-resistant lysogenic and non-lysogenic substrains were produced from two Staphylococcus aureus L-form strains lysogenic for different prophages, namely, EMT-L (prophage alpha) and 209P (prophage beta) . Cells of these L-form substrains were fused in various combinations using polyethylene glycol (PEG), and the frequency of recombinants selected as double resistance to both SM and EM and the prophage types of these recombinants were examined . In all the combinations, the frequency of recombinants was greater when the cells were treated with PEG than when they were not, and the difference was statistically significant (p less than 0.01) in 13 combinations . Combination between the lysogenic SM-resistant EMT-L substrain {EMT(Smr-alpha)} and lysogenic EM-resistant 209P-L substrain {209P(Emr-beta)} and the reverse combination, between 209P(Smr-beta) and EMT(Emr-alpha), resulted in a majority of recombinants harboring prophage beta . The former combination yielded recombinants that all held both prophage alpha and beta.

Biochem J, 1986 Jun 1, 236(2), 389 - 95
Identification of the sites in opsin modified by photoactivated azido{125I}iodobenzene; Davison MD et al.; Opsin labelled with photoactivated 1-azido-4-{125I}iodobenzene was proteolysed in situ with Staphylococcus aureus V8 proteinase to yield two radioactive membrane-bound fragments . These were separated, cleaved with CNBr and the resultant peptides sequenced in order to locate the radiolabelled residues . In the whole molecule, there was clear evidence for modification of at least 20 sites, identified as derivatives of cysteine, tryptophan, tyrosine, histidine and lysine residues . The probe primary reacted, therefore, with nucleophilic substituents . The positions of the modified sites relative to the confines of the phospholipid bilayer were consistent with all other studies on the disposition of the polypeptide chain . The location of these sites substantiated an earlier suggestion that not all the transmembrane segments should be regarded as continuous regular alpha-helices.

Brain Res, 1986 May 14, 373(1-2), 1 - 14
Production and characterization of polyclonal and monoclonal antibodies to rat brain L-glutamate decarboxylase; Wu JY et al.; Specific monoclonal and polyclonal antibodies to rat brain glutamate decarboxylase (GAD) were produced and characterized . Polyclonal antibodies against GAD were raised in rabbits by injecting a total of 70-210 micrograms of purified GAD i.m . The specificity of anti-GAD serum was established from a variety of tests including Ouchterlony immunodiffusion, immunoelectrophoresis, immunoprecipitation, dot immunoassay, ELISA tests and Western immunoblottings . In immunodiffusion and immunoelectrophoresis tests using partially purified GAD preparations and anti-GAD serum a single, sharp precipitin line corresponding to GAD activity was obtained . Quantitative immunoprecipitation of GAD activity was achieved using anti-GAD IgG and Staphylococcus aureus . Specificity of the antiserum was further indicated from a dot immunoassay and ELISA tests in which the intensity of the reaction product was proportional to the amount of GAD protein present . In the Western immunoblotting experiments using partially purified GAD preparations only two protein bands corresponding to the position of the two subunits of GAD were stained by anti-GAD IgG, further supporting the specificity of polyclonal antibodies against GAD . In addition to polyclonal antibodies, several specific GAD-antibodies-producing clones were also obtained by the hybridoma technique . The specificity of monoclonal antibodies against GAD were established from the following criteria: positive on ELISA test using homogeneous GAD as antigen; formation of GAD--anti-GAD IgG complex as indicated from gel filtration chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis; and specific recognition of GAD subunit in a partially purified GAD preparation in Western immunoblotting test . Monoclonal antibodies were further characterized by immunohistochemical localization of known GABAergic neurons and their processes in the cerebellum and retina.

Presse Med, 1986 May 10, 15(19), 876 - 8
{Mycotic aneurysm after catheterization of the umbilical artery}; Dubos JP et al.; The finding of an abdominal mass in an 18-month old infant ultimately led to the diagnosis of mycotic aneurysm of a common iliac artery . The lesion was resected and the vessel was ligated . The short and long-term outcome was favourable . The child had been operated upon for cervical teratoma and had undergone catheterization of the umbilical artery complicated with Staphylococcus aureus infection . The presence of a mycotic aneurysm must be suspected in infants with a history of umbilical artery catheterization or neonatal staphylococcal infection, or presenting with a posterior mediastinal or abdominal mass, or arterial hypertension . The vessel most commonly involved is the aorta . Surgical resection, when performed, results in cure . The present case is remarkable on three points: the lesion involved an iliac artery, the diagnosis was delayed and calcifications were present around the aneurysm.

Minerva Med, 1986 May 7, 77(19), 811 - 4
{Pathogenic microorganisms on the pharyngeal swabs from hospital personnel}; Vacca G et al.; After a survey of the literature concerning nosocomial infections, 593 healthy human subjects belonging to the "Ospedale S . Croce Cuneo" staff, have been investigated for the presence of Staphylococcus aureus and other pathogenic bacteria in their oropharyngeal secretions.

J Biol Chem, 1986 May 5, 261(13), 5777 - 83
Characterization of hormone and protein release from alpha-toxin-permeabilized chromaffin cells in primary culture; Bader MF et al.; Addition of Staphylococcus aureus alpha-toxin to adult bovine chromaffin cells maintained in primary culture causes permeabilization of cell membrane as shown by the release of intracellular 86Rb+ . The alpha-toxin does not provoke a spontaneous release of either catecholamines or chromogranin A, a protein marker of the secretory granule, showing the integrity of the secretory vesicle membrane . However the addition of micromolar free Ca2+ concentration induced the co-release of noradrenaline and chromogranin A . In alpha-toxin-treated cells, the released chromogranin A could not be sedimented and lactate dehydrogenase was still associated within cells, which provides direct evidence that secretory product is liberated by exocytosis . By contrast, permeabilization of cells with digitonin caused a Ca2+-dependent but also a Ca2+-independent release of secretory product, a dramatic loss of lactate dehydrogenase, as well as release of secretory product in a sedimentable form . Ca2+-dependent exocytosis from alpha-toxin-permeabilized cells required Mg2+-ATP and did not occur in the presence of other nucleotides . Thus alpha-toxin is a convenient tool to permeabilize chromaffin cells, and has the advantage of keeping intracellular structures, specifically the exocytotic machinery, intact.

Am J Dis Child, 1986 May, 140(5), 459 - 61
The value of skin biopsies in febrile, neutropenic, immunocompromised children; Allen U et al.; We assessed the value of 41 skin biopsy specimens obtained from 32 immunocompromised patients with fever and neutropenia . Fifty-six percent (23/41) of these biopsy specimens resulted in a specific diagnosis . These diagnoses included infection in 12 cases, graft-vs-host disease in nine, leukemic infiltrate in one, and drug reaction in one . The remaining 18 biopsy specimens showed either nonspecific changes or normal tissue . The infectious agents identified included Aspergillus in nine patients, Candida in three patients, and Staphylococcus aureus in one patient . As a result of the biopsy findings, therapy was altered in 49% (20/41) of the cases . The most frequent therapeutic change was initiation of amphotericin B for a demonstrated fungal infection . We found skin biopsies to be a valuable diagnostic tool in the assessment of the febrile, neutropenic, immunocompromised patient with skin lesions.

Arch Pathol Lab Med, 1986 May, 110(5), 442 - 4
Umbilical artery catheterization complicated by multiple mycotic aortic aneurysms; Rabin E et al.; A premature infant had three pseudoaneurysms of the thoracic and abdominal aorta secondary to umbilical artery catheterization and sepsis . The infant had septicemia as the direct result of bacterial contamination of an umbilical artery catheter with Staphylococcus aureus . The thoracic pseudoaneurysm caused massive hemothorax and the infant's death . The upper abdominal aortic aneurysm developed at the level of the renal arteries and caused decreased left renal blood flow and renal hypoplasia . The lower abdominal aneurysm involved the right iliac artery and was complicated by mural thrombosis and ischemia of the right leg . To our knowledge, this is the first published case of multiple mycotic aortic aneurysms after umbilical artery catheterization.

Antimicrob Agents Chemother, 1986 May, 29(5), 930 - 2
Determination of aminoglycoside resistance in Staphylococcus aureus by DNA hybridization; Dickgiesser N et al.; A method is described for identification of the genes conferring aminoglycoside resistance in Staphylococcus aureus by dot-blot and Southern blot techniques . As radioactive probes, fragments of plasmids pAT48, pUBH2, and pH13, carrying the genes for an aminocyclitol-3'-phosphotransferase, an aminocyclitol-4'-adenylyltransferase, and an aminocyclitol-2''-phosphotransferase-aminocyclitol-6'-acetyltransferase, respectively, were used.

Zh Mikrobiol Epidemiol Immunobiol, 1986 May, (5), 67 - 73
{Mitogenic activity of the antigens isolated from different Staphylococcus aureus strains in relation to mouse and guinea pig splenocytes}; Khorobrykh VV et al.; The mitogenic effect of corpuscular antigens with respect to the splenocytes of animals was found to depend on the strain of Staphylococcus aureus . The maximum synthesis of DNA in the cells was induced by corpuscular antigen Smith and the minimum synthesis, by Wood-46 . The synthesis of DNA was activated in both B- and T-splenocytes in response to corpuscular antigens Wood-46, Cowan-1 and Smith, as well as to the cell wall and protein A . Peptidoglycan produced a mitogenic effect only in B-lymphocytes, and teichoic acid showed no mitogenic activity in mouse splenocytes . The mitogenic effect of staphylococcal antigens on splenocytes depended on the dose of the antigen and the time of cultivation . After 48-hour cultivation the incorporation of 3H-thymidine into the DNA of mouse cells was 5 times higher than into the DNA of guinea-pig cells . The optimum mitogenic dose in thymectomized BALB/c mice with respect to splenocytes was higher than in normal BALB/c mice practically by one order.

Am J Vet Res, 1986 May, 47(5), 1139 - 41
Lymphocyte blastogenesis and neutrophil function in cattle persistently infected with bovine viral diarrhea virus; Roth JA et al.; Neutrophil function and mononuclear cell proliferative responses to mitogens were determined in healthy cattle and in cattle persistently infected with bovine viral diarrhea (BVD) virus . Uptake of {3H}thymidine by resting and mitogen-stimulated peripheral blood mononuclear cells was significantly lower in cattle persistently infected with BVD virus than in healthy cattle . Neutrophils from cattle persistently infected with BVD virus had significantly impaired capability to ingest Staphylococcus aureus, but were normal in respect to random migration under agarose, cytochrome C reduction, iodination, and antibody-dependent cell-mediated cytotoxicity . Impairment of neutrophil function in cattle persistently infected with BVD virus differs from impairment of neutrophil function reported in healthy cattle mounting an immune response to recent BVD virus infection.

J Biomed Mater Res, 1986 May-Jun, 20(5), 565 - 77
Bacterial inhibition by electrical activation of percutaneous silver implants; Spadaro JA et al.; Percutaneous silver wire implants were looped through the dorsal skin of rats and inoculated with Staphylococcus aureus to test the effect on bacteria in the tract . The silver was activated with four brief daily applications of anodic microcurrent . Contralateral 316L stainless steel implants, identically inoculated, served as controls . Cultures from the silver tracts showed a marked reduction or elimination of bacteria, which persisted for the 3-week study period . In tracts with colonization established for 1 week, subsequent electrical activation of the silver also suppressed the bacteria . Inflammatory reactions at 3 weeks were mild at both the silver and stainless implants and no giant cells or toxicity were seen . This suggests that electrically activated silver may be useful in preventing or treating infection at percutaneous devices.

Br Heart J, 1986 May, 55(5), 497 - 9
Echocardiographic demonstration of free wall vegetative endocarditis complicated by a pulmonary embolism in a patient with ventricular septal defect; Zijlstra F et al.; A defect in the muscular part of the interventricular septum in a 19 year old man was complicated by infective endocarditis caused by Staphylococcus aureus . The lesion was a large right ventricular free wall vegetation which embolised to the lungs . The vegetation was displayed by cross sectional echocardiography, which also confirmed the clinical diagnosis of ventricular septal defect . This case confirms the concept that the jet stream causes endocarditis at its point of impact . After six weeks' treatment with antibiotics the ventricular septal defect was repaired at operation.

J Med Chem, 1986 May, 29(5), 661 - 4
Synthesis of delta 3-1-methylene-1-carbacephems; Herdewijn P et al.; The total synthesis of (+/-)-1-methylene-2,2- dimethyl-7-amino-1-carbacephem-4-carboxylic acid (1) is described . The reaction scheme was essentially that described by Christensen et al . for the synthesis of (+/-)-1-carbacephems . In vitro antibacterial activities of the 7-phenoxyacetyl and 7-D-alpha-phenylglycyl derivatives of 1 were compared with those of 7-(phenoxyacetamido)desacetoxycephalosporanic acid and cefalexin . Derivatives of 1 were 2-4 times less active against most of the sensitive organisms than the corresponding 7-aminodesacetoxycephalosporanic acid analogues . The activity of the 7-D-alpha-phenylglycyl derivative of 1 however was about 20 times lower than that of cefalexin when measured against Staphylococcus aureus ATCC 6538P.

J Infect Dis, 1986 May, 153(5), 956 - 9
Clindamycin treatment of experimental chronic osteomyelitis due to Staphylococcus aureus; Norden CW et al.; Clindamycin was used alone for treatment of experimental osteomyelitis due to Staphylococcus aureus in rabbits . Treatment with 30 mg/kg of body weight three times a day for 14 days was ineffective in sterilizing infected rabbit bones . In contrast, when given for 28 days, clindamycin sterilized the infected bones of 16 (84%) of 19 rabbits treated . Only one of 14 isolates of S . aureus from rabbits treated for two weeks developed resistance to clindamycin (minimal inhibitory concentration, greater than 100 micrograms/ml); none of three isolates from rabbits in which treatment failed in the four-week treatment group showed resistance to clindamycin . The results of four weeks of treatment with clindamycin for chronic experimental staphylococcal osteomyelitis were significantly better than those obtained with any other single agent used in prior studies and were generally as good as those with combination therapy that included rifampin.

J Antimicrob Chemother, 1986 May, 17(5), 571 - 4
Probable chromosomal mutation to resistance to all aminoglycosides in Staphylococcus aureus selected by the therapeutic use of gentamicin: a preliminary report; Heritage J et al.; A patient suffering from burns was treated with gentamicin: subsequently, a 'methicillin-resistant' strain of Staphylococcus aureus (B27), was isolated which was highly resistant to this antibiotic and all other aminoglycosides . In all other respects this isolate resembled the gentamicin-sensitive 'methicillin-resistant' S . aureus prevalent on the Yorkshire Regional Burns Unit . Subsequently, fusidic acid treatment was substituted for gentamicin therapy, and a fusidic acid-resistant variant was isolated from the same patient . This article reports a preliminary characterization of these isolates.

Br J Hosp Med, 1986 May, 35(5), 312, 314, 318 - 20
Staphylococcal infections in hospital; Shanson DC; Staphylococcus aureus is a major cause of surgical sepsis and septicaemia . The prevention and control of outbreaks of hospital infection caused by methicillin-resistant strains has become increasingly important in recent years . Staph . epidermidis has also emerged as a "problem organism" often causing serious infection in patients with prosthetic implants.

J Clin Microbiol, 1986 May, 23(5), 916 - 9
Evaluation of rapid coagulase methods for the identification of Staphylococcus aureus; Berke A et al.; Four rapid latex agglutination assays, StaphAurex (Wellcome Diagnostics, Research Triangle Park, N.C.), Bacto Staph (Difco Laboratories, Detroit, Mich.), SeroSTAT (Scott Laboratories, Inc., Fiskeville, R.I.), Veri-Staph (Zeus Technologies, Raritan, N.J.), and two hemagglutination tests, Staphyloslide (BBL Microbiology Systems, Cockeysville, Md.) and Hemastaph (Remel, Lenexa, Kans.), were compared with the conventional slide coagulase, tube coagulase (TC), and thermonuclease (TNase) tests for the identification of Staphylococcus aureus . A total of 118 clinical isolates of S . aureus (52 methicillin resistant), 50 S . epidermidis, 5 S . capitis, 2 S . hominis, 3 S . simulans, 6 S . saprophyticus, and 2 S . warneri were tested . The slide coagulase, TC and TNase tests detected 115 (97.5%), 117 (99.2%), and 118 (100%) of the S . aureus isolates, respectively . All showed 100% specificity . The StaphAurex, Veri-Staph, Staphyloslide, Hemastaph, SeroSTAT, and Bacto Staph assays correctly identified 117 (99.2%), 117 (99.2%), 116 (98.3%), 110 (93.2%), 108 (91.5%), and 107 (90.7%) of the S . aureus isolates, respectively . For methicillin-resistant S . aureus isolates, StaphAurex, Veri-Staph, Staphyloslide, Hemastaph, SeroSTAT, and Bacto Staph showed 1 (2%), 1 (2%), 2 (4%), 7 (13.5%), 7 (13.5%), and 8 (15.4%) false-negative results, respectively . All the commercial agglutination assays demonstrated false-positive results with strains of S . capitis, S . saprophyticus and S . warneri . The overall accuracy of the commercial agglutination assays compared with TC and TNase ranged from 90.7 to 99.2% . We recommend that negative reactions with the rapid commercial test kits for methicillin-resistant Staphylococcus isolates be confirmed with the TC or TNase test.

Infect Control, 1986 May, 7(5), 263 - 7
Category 1, 2, 3 and 4: a procedure-oriented isolation system; Gilmore DS et al.; A procedure-oriented isolation system, Category 1,2, 3, and 4, was introduced at a 547-bed, acute and rehabilitative medical center . The system consisted of four categories of isolation which followed a numerical sequence that represented the necessary attire needed to complete the procedure . After 1 year of use, personnel compared the procedure-oriented system with the previously-used system (Strict, Respiratory, Wound and Skin, Enteric, and Limited Barrier) . Personnel found the procedure-oriented system easier to understand (84%) and follow (83%) . Seventy-six percent felt their isolation techniques had improved with the new system . A reduction in the cross-infection rate with methicillin-resistant Staphylococcus aureus did coincide with the use of the new isolation system, however, no causal relationship was established . The Category 1, 2, 3, and 4 isolation system was well received by personnel and was found to be an effective alternative to the previous, more complicated system used in this setting . Further evaluation of this system in other settings would seem warranted.

Am J Med, 1986 May, 80(5), 770 - 6
Methicillin-resistant Staphylococcus aureus bacteremia linked to intravenous drug abusers using a "shooting gallery"; Craven DE et al.; Over a 15-month period, seven intravenous drug abusers had 10 admissions because of bacteremia due to methicillin-resistant Staphylococcus aureus . Seven episodes of probable bacterial endocarditis occurred in four patients; one patient had septic thrombophlebitis and two had soft tissue infections . All seven patients patronized a local "shooting gallery" where paraphernalia were provided and drugs were often administered by a "street doctor." All isolates were phage type 29/77/83A/84/85 and demonstrated resistance only to methicillin, oxacillin, and penicillin . This strain of methicillin-resistant S . aureus has a phage type and antibiogram that is distinct from nosocomial methicillin-resistant S . aureus and was probably acquired by intravenous drug abusers during visits to the "shooting gallery" . The "shooting gallery" is an integral part of the drug culture and a likely source for the transmission of antibiotic-resistant organisms.

Mol Gen Mikrobiol Virusol, 1986 May, (5), 45 - 6
{The use of a dot immunoenzyme method for detection of viral antigens}; Tarasishin LA; The technique of dot immunoenzyme detection has been elaborated for viral antigens identification . The investigated samples (extracts of infected cells, fractions obtained during viruses and viral proteins purification) are placed in nitrocellulose filters, the free binding sites are blocked and then treated with specific immune serum . The formed antigen-antibody complexes are detected using antispecies immunoglobulins or protein A from Staphylococcus aureus, conjugated with horseradish peroxidase . The brown dots appear at samples location containing the viral antigens . Sensitivity of the technique is 1 ng of protein per sample as tested using adenoviral antigens.

Pathol Biol (Paris), 1986 May, 34(5), 465 - 9
{Treatment of moderate or severe infections using imipenem/cilastatin . 41 cases based on a multicenter protocol}; Stahl JP et al.; Imipenem and cilastatin in combination have a broad spectrum in vitro with a strong killing activity on most bacteria . Using a multicenter study design, we investigated 41 patients with moderate or severe infections: septicemia in 18 cases (Gram negative rods in 10, Gram positive cocci in 7 and combination of both in 1), pneumonia in 7, osteitis in 4, soft tissue infection in 7, infection of the genitourinary tract in 6 and miscellaneous infections in the remaining cases (1 abscess of the pancreas, 1 typhoid fever, 1 presumptive endocarditis) . All of the bacteria were susceptible to imipenem/cilastatin: MICs ranged from 0.02 to 0.8 mg/l and MBCs from 0.015 to more than 10 mg/l . All patients except one recovered or improved under imipenem/cilastatin . The patient who failed to respond had septicemia due to a methicillin-resistant Staphylococcus aureus with a MBC and MIC above 10 and 0.5 mg/l respectively . Tolerance was outstanding: only 4 patients had adverse effects requiring withdrawal of the drug.

Pathol Biol (Paris), 1986 May, 34(5), 357 - 9
{Diffusion of fosfomycin into the human and rabbit eye (aqueous humor and vitreous body)}; Denis F et al.; The intraocular distribution of fosfomycin was studied in 32 patients undergoing cataract surgery and in 8 rabbits after experimental infection of one eye by Staphylococcus aureus . Concentrations found 1 to 6 hours after termination of a 4 g fosfomycin infusion ranged from 14 to 18.8 mg/l in the aqueous humor and 8 to 12.5 mg/l in the vitreous body . These levels are higher than the MICs for 80 to 90% of the bacteria responsible for endophthalmitis . In each rabbit, the fosfomycin concentration in the infected eye as compared to the healthy eye was increased 2.5 to 5--fold for the aqueous humor and 4.9 to 19.2--fold for the vitreous body . Fosfomycin, in association with a third generation cephalosporin (ceftriaxone) or one of the new quinolones (pefloxacin) can be recommended for the prevention or early treatment of endophthalmitis.

J Dairy Res, 1986 May, 53(2), 197 - 202
Determinants of bacterial replication rates in mastitic whey; Mattila T et al.; Bacterial growth was measured by a turbidimetric microtechnique in the whey of milk samples from quarters of cows with subclinical mastitis . Samples were grouped according to bacterial isolates recovered and the effects of bacterial species and whey on bacterial growth rates were analysed . Different strains of bacteria and different whey samples gave highly significant differences in bacterial replication rates . Except for penicillin-resistant Staphylococcus aureus, bacteria grew better in whey from mastitic milk where the inflammation was caused by the same bacterial species than in other mastitic milk samples . Inflammation caused by major pathogens generally enhanced the growth in whey of any type of major pathogen . Since mastitis pathogens showed enhanced growth in whey prepared from the same milk from which they were isolated, specific antibacterial factors in the whey did not appear to restrict bacterial growth in whey . The nutritional quality of the medium seems to be the important determinant of bacterial growth.

Hum Immunol, 1986 May, 16(1), 1 - 13
Activation of human B cells: involvement of surface immunoglobulin as evidenced by two biochemically distinct types of response to Staphylococcus aureus; Scholten P et al.; If activation of human B cells by Staphylococcus aureus proceeds through interaction of surface immunoglobulin with Staphylococcal protein A, then immunoglobulins should be produced that are capable of binding to protein A as a consequence of such stimulation . In the present report it is shown that two biochemically distinct types of response to S . aureus are demonstrable in human peripheral blood lymphocytes . Two types of IgM are produced: IgM capable of binding to protein A, and IgM that does not bind and can be recovered by immunoprecipitation with anti-Ig antibodies . Cloned cell lines produce one of either type of Ig, but not both . Therefore, interaction of protein A with Ig alone cannot account for the stimulatory properties of S . aureus . When S . aureus is used in conjunction with pokeweek mitogen, a synergistic effect between the two mitogens is seen . Under conditions of optimal synergistic stimulation, the increase in immunoglobulin production is seen virtually exclusively in the category of molecules capable of binding to protein A . These results offer strong support for a model where optimal differentiation of human B cells to plasma cells is contingent upon receiving at least two signals: one signal is delivered to the surface immunoglobulin, and one signal delivered to the B cell in a T cell and/or monocyte dependent fashion (B cell growth and differentiation factors).

Immunology, 1986 May, 58(1), 37 - 41
Characteristics of human B cells responsive to the T-independent mitogen Branhamella catarrhalis; Calvert JE et al.; Non-T cells from tonsil or blood were fractionated according to buoyant density, isotype of surface immunoglobulin, or the ability to form rosettes with mouse erythrocytes . Each fraction was tested for the ability to proliferate in response to B . catarrhalis (Bc) and, for comparison, Staphylococcus aureus Cowan 1 (SAC) or an MLR supernatant (TF) . Cells in all density fractions responded to Bc, the greatest response occurring in the high-density cell fraction . SAC could similarly induce proliferation in high-density cells, in contrast to TF which preferentially activated cells in the low-density fraction . When cells were fractionated by rosetting with mouse erythrocytes, both fractions (MRBC-R+ and MRBC-R-) responded to Bc and to SAC, whereas the greatest response to TF occurred in the MRBC-R- cell fraction . Depletion of sIgD+ sIgM+ cells almost completely abolished the response to Bc, suggesting that responsive cells express both these classes of immunoglobulin on their membrane . Furthermore inclusion of anti-delta antibodies in cultures resulted in failure to proliferate in response to Bc . These data strongly suggest that Bc, like SAC but in contrast to TF, is able to stimulate proliferation in cells with the characteristics of resting B cells, i.e., high-density, sIgD+ cells which form rosettes with MRBC . This may be related to the fact that Bc, like SAC, is able to bind to human immunoglobulins.

Immunology, 1986 May, 58(1), 31 - 5
Direct induction of human B-cell differentiation by recombinant interleukin-2; Romagnani S et al.; Recombinant interleukin-2 (rIL-2) induced highly purified human tonsillar B cells to differentiate into immunoglobulin (Ig)-producing cells in vitro . The B-cell response was not due to rIL-2-contaminating substances, but reflected the activity of IL-2 itself, since it was inhibited by addition to the cultures of anti-TAC monoclonal antibody . The rIL-2-induced B-cell response was apparently not mediated by factors released by residual T cells present in B-cell suspensions at undetectable levels, since supernatants (SN) from unstimulated autologous T cells cultured at concentrations even much higher than those possibly contaminating B-cell suspensions did not induce any detectable Ig production . In addition, the Ig production by B cells cultured with SN prepared from high numbers of autologous T cells stimulated with rIL-2, as well as from allo-activated or mitogen-stimulated T cells, was of the same magnitude as the Ig production resulting from direct addition of rIL-2 concentrations comparable with those present in the supernatants . After centrifugation on Percoll density gradients, most of the tonsillar B cells responsive to rIL-2 were recovered in the lower density cell fraction containing a number of larger activated B cells . Moreover, B-cell enriched suspensions from peripheral blood (PB) (which usually contains a lower number of in vivo activated B cells than tonsil) showed poor or no response to rIL-2 alone, but displayed significant Ig production when rIL-2 was added to the cultures in the presence of Staphylococcus aureus Cowan I (SAC) bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

J Immunol, 1986 May 1, 136(9), 3311 - 6
Purification and characterization of a unique human interleukin 1 from the tumor cell line U937; Knudsen PJ et al.; Interleukin 1 (IL 1) produced by a human tumor cell line was purified to homogeneity by a three-step chromatographic method and was tested in various assays for multiple biologic properties . The purified IL 1 stimulated the proliferative response of the D10.G4.1 cell line, a mouse IL 1 indicator T cell; caused the release of prostaglandin E2 and prostacyclin from cultured human foreskin fibroblasts and from primary human umbilical vein endothelial cells; and elicited characteristic endogenous pyrogen fever in rabbits . To stimulate IL 1 production, the histiocytic lymphoma cell line U937 was incubated with the exotoxin from toxic shock strains of Staphylococcus aureus . Supernatants from stimulated U937 cells were concentrated, and were applied to a reverse-phase HPLC column . IL 1 activity was eluted from the column at high acetonitrile concentration . Subsequent chromatography over hydroxyapatite yielded a single IL 1 species with a pI of 5.5 . IL 1 was then purified to homogeneity by gel exclusion HPLC migrating as a 14 kDa species . The molecular size was confirmed by SDS-PAGE and was visualized as a single molecule by silver staining; biologic activity was recovered from the same region of the gel . Limited N-terminal sequence analysis suggested some homology to the pI 7 form of the human blood monocyte IL 1 . The pI 5.5 IL 1 produced by U937 cells was only partially neutralized with anti-human monocyte IL 1 antibody, suggesting that U937-derived IL 1 is structurally related to one of the molecularly cloned IL 1 species . IL 1 from stimulated U937 cells possesses the functional characteristics of monocyte IL 1 but may represent a structurally unique IL 1 species, as determined by sequence analysis, size, and antibody reactivity.

Proc Natl Acad Sci U S A, 1986 May, 83(9), 3008 - 11
Mapping of the alpha-bungarotoxin binding site within the alpha subunit of the acetylcholine receptor; Neumann D et al.; Synthetic peptides and their respective antibodies have been used in order to map the alpha-bungarotoxin binding site within the alpha subunit of the acetylcholine receptor . By using antibodies to a synthetic peptide corresponding to residues 169-181 of the alpha subunit, we demonstrate that this sequence is included within the 18-kDa toxin binding fragment previously reported . Furthermore, the 18-kDa fragment was also found to bind a monoclonal antibody (5.5) directed against the cholinergic binding site . Sequential proteolysis of the acetylcholine receptor with trypsin, prior to Staphylococcus aureus V8 protease digestion, resulted in a 15-kDa toxin binding fragment that is included within the 18-kDa fragment but is shorter than it only at its carboxyl terminus . This 15-kDa fragment therefore initiates beyond Asp-152 and terminates in the region of Arg-313/Lys-314 . In addition, experiments are reported that indicate that in the intact acetylcholine receptor, Cys-128 and/or Cys-142 are not crosslinked by disulfide bridges with any of the cysteines (at positions 192, 193, and 222) that reside in the 15-kDa toxin binding fragment . Finally, the synthetic dodecapeptide Lys-His-Trp-Val-Tyr-Tyr-Thr-Cys-Cys-Pro-Asp-Thr, which is present in the 15-kDa fragment (corresponding to residues 185-196 of the alpha subunit) was shown to bind alpha-bungarotoxin directly . This binding was completely inhibited by competition with d-tubocurarine.

Proc Natl Acad Sci U S A, 1986 May, 83(9), 2822 - 6
Widespread occurrence of "87 kDa," a major specific substrate for protein kinase C; Albert KA et al.; An 87-kDa phosphoprotein, identified previously as a major, specific substrate for Ca2+/phospholipid/diacylglycerol-dependent protein kinase (protein kinase C) in broken cell preparations from rat brain, has been characterized with respect to its species, tissue, and subcellular distribution . A similar protein was present in monkey, human, mouse, and bovine brain and in Torpedo californica electric organ . The protein was also identified in a variety of nonneuronal rat and bovine tissues . The rat protein had an apparent molecular mass 4-7 kDa lower, and was slightly more acidic, than the protein in bovine tissues . The 87-kDa proteins from various bovine tissues were identical by the following criteria: each was phosphorylated by exogenous protein kinase C, was of comparable molecular mass, generated multiple spots within the pH range of 4.4-4.9 upon isoelectric focusing, yielded identical patterns upon digestion with Staphylococcus aureus V8 protease, and was recognized by a specific 87-kDa antiserum . The relative concentrations of the 87-kDa protein in bovine tissues were highest in brain, spleen, and lung, moderate in testis, pancreas, adrenal, kidney, and liver, and lowest in heart and skeletal muscle . In the brain, the 87-kDa protein was concentrated in the synaptosomal membrane and in the cytosol . The membrane-bound protein was extractable with nonionic detergents but not with NaCl . This species, tissue, and subcellular distribution of the 87-kDa protein is similar to that of protein kinase C.

Can J Microbiol, 1986 May, 32(5), 442 - 5
Antibacterial activity of BCG-induced, interferon-containing sera; Moulton RG et al.; Sera from mice which have been vaccinated with BCG and challenged with old tuberculin contain gamma interferon . These same sera also express antibacterial activity . Using Staphylococcus aureus we demonstrated that its growth was inhibited at dilutions of sera as high as 1:320 . A 4% concentration of sera reduced the growth rate of the S . aureus from 1.6 to 0.6 doubling times per hour . The activity was stable at 56 degrees C but destroyed by 80 degrees C . It was nondialysable and destroyed by acid conditions (pH 2.0) and by the proteolytic enzymes trypsin and chymotrypsin . Antibodies to gamma interferon neutralized the antiviral activity but not the antibacterial activity . Mitogen-induced and virus-induced interferons did not have activity . We subsequently demonstrated that the factor could be induced in mice using BCG without the secondary old tuberculin challenge . No gamma interferon was found in the sera of mice given BCG without old tuberculin . These findings indicate that the antibacterial activity of these sera is not dependent on the presence of gamma interferon . We will continue to work to characterize and identify the antibacterial component in these sera.

AJNR Am J Neuroradiol, 1986 May-Jun, 7(3), 395 - 402
Experimental Staphylococcus aureus brain abscess; Enzmann DR et al.; The virulent organism Staphylococcus aureus produced brain abscesses that were quantitatively and qualitatively different from those caused by less virulent organisms . S . aureus abscesses created larger lesions, as earlier ependymitis, delayed progress toward healing, and caused areas of inflammatory escape outside the collagen capsule . Imaging tests revealed similar findings: the abscesses were larger, had more extensive central necrosis, and showed earlier evidence of ependymitis . This virulent organism also demonstrated that white matter is more susceptible than overlying gray matter to destruction by infection . The pattern of spread and other histologic findings suggest that collagen capsule formation has less of an infection "containment" function than was previously thought.

Clin Immunol Immunopathol, 1986 May, 39(2), 285 - 97
Effects of anti-IgM on mitogen-induced proliferation of human B-lymphocyte malignancies; Baeker TR et al.; Therapeutic trials of anti-immunoglobulin antibody have produced a wide range of responses in attempts to control the growth of human B lymphoid neoplasms . This variability might reflect differences in intrinsic functional characteristics of malignant B lymphocytes that determine susceptibility to anti-immunoglobulin-mediated regulation of growth . To characterize B-lymphocyte malignancies, tissue samples from 24 patients were studied during short-term culture in vitro . Malignant B lymphocytes were stimulated to proliferate by the T-independent mitogens lipopolysaccharide, cytochalasin B, and Staphylococcus aureus that bears protein A . The effects of monoclonal mouse anti-human IgM on mitogen-induced malignant lymphocyte proliferation were then assessed . Mitogen-induced responses of malignant lymphocytes from three patients were abrogated by 2 micrograms/ml monoclonal anti-human IgM . Proliferation was also abrogated by polyclonal goat anti-IgM antiserum but proliferative responses were not affected by control monoclonal antibody . Further study showed that anti-immunoglobulin-mediated inhibition of proliferation was not dependent on Fc-determined interactions, nor was it dependent on the presence of T lymphocytes . These results indicate that a subset of human B-lymphocyte malignancies are susceptible to inhibition of proliferation mediated by anti-IgM.

J Immunol, 1986 May 1, 136(9), 3288 - 91
Requirement of macrophages (monocytes) for the induction of interleukin 2 receptors on B lymphocytes; Shirakawa F et al.; The role of macrophages (monocytes) for the induction of interleukin 2 receptors (IL 2R) on human B lymphocytes was studied by a direct immunofluorescence method with the use of a fluoresceinated anti-IL 2R monoclonal antibody and a flow cytofluorometer . Highly purified B lymphocytes alone did not induce IL 2R on their surface by stimulation with Staphylococcus aureus Cowan I, anti-mu antibody, or pokeweed mitogen . However, the addition of monocytes successfully induced IL 2R on B lymphocytes stimulated with these mitogens in a dose-dependent manner . Interleukin 1 (IL 1) produced by monocytes could partially replace the accessory function of monocytes . In accordance with these results, the proliferation of B lymphocytes and the differentiation to immunoglobulin-producing cells in response to IL 2 were also dependent on monocytes or IL 1 . These results suggest that the accessory function of macrophages for IL 2-induced B cell activation is primarily on the induction of IL 2R on B lymphocytes.

Acta Paediatr Scand, 1986 May, 75(3), 444 - 51
Phagocytosis-associated oxidative metabolism in human milk macrophages; Speer CP et al.; Evidence exists that human milk macrophages, which are the most abundant cells in milk, play an important role in the protection of the newborn infant against infection . We investigated the oxidative metabolism of milk macrophages by measuring luminol-dependent chemiluminescence, generation of superoxide anion (O2-) and production of hydrogen peroxide (H2O2) in the resting state and after stimulation with opsonized zymosan or phorbol myristate acetate (PMA) . Generally, macrophages generated luminol-dependent luminescence, O2- and H2O2 with either stimulus to a similar extent as did blood monocytes . In addition, macrophages killed Escherichia coli and Staphylococcus aureus as effectively as did blood monocytes (approximately 75%, 120 min) . Acidification of macrophages in milk (pH 1, 30 min) only slightly reduced their PMA-stimulated production of oxygen radicals . Bacterial killing by macrophages preincubated at pH 1 was about 70% that from controls maintained at pH 7 . When macrophages were cultured for several days in endotoxin-free medium, their ability to produce oxygen metabolites declined . By continuous treatment with bacterial LPS (10 ng/ml), milk-macrophages could be "primed" to release large amounts of oxygen intermediates . In addition, the O2- response of macrophages cultured without bacterial products could be partially restored by the addition of LPS to the culture . We conclude that milk macrophages are capable of releasing large amounts of oxygen metabolites, and could contribute to the protection of neonates against bacterial and fungal infections.

J Clin Microbiol, 1986 May, 23(5), 832 - 9
The role of beta-lactamase in staphylococcal resistance to penicillinase-resistant penicillins and cephalosporins; McDougal LK et al.; We showed that most Staphylococcus aureus strains that have borderline or intermediate susceptibility to the penicillinase-resistant penicillins (PRPs) react this way because of the activity of their beta-lactamase on these antimicrobial agents . These strains produced large amounts of staphylococcal beta-lactamase that rapidly hydrolyzed penicillin and partially hydrolyzed the PRPs . Susceptibility to hydrolysis was penicillin greater than oxacillin greater than cephalothin greater than methicillin . The borderline results and the hydrolysis could be prevented by the beta-lactamase inhibitors clavulanic acid and sulbactam . For intrinsically methicillin-resistant (heteroresistant) S . aureus, the inhibitors reduced the penicillin MICs, but the strains remained resistant to all the beta-lactam antimicrobial agents, including penicillin . We conclude that the borderline in vitro susceptibility or resistance to PRPs in most of these S . aureus strains is mediated by beta-lactamase and they are not heteroresistant or intrinsically resistant . We do not know whether this in vitro resistance is expressed clinically.

J Clin Oncol, 1986 May, 4(5), 798 - 804
Impaired in vitro polymorphonuclear function secondary to the chemotherapeutic effects of vincristine, adriamycin, cyclophosphamide, and actinomycin D; Cairo MS et al.; The present study investigated the in vitro effect of four different chemotherapeutic agents, namely, cyclophosphamide (CTX), vincristine (VCR), Adriamycin (Adria Laboratories, Columbus, Ohio) (ADR), and actinomycin D (ACT-D) on human polymorphonuclear leukocyte (PMN) function . Human PMNs suspended in phosphate-buffered saline (PBS) at 1 X 10(7) cells/mL were incubated with increasing concentrations of CTX (0, 10(-5), 10(-4), 10(-3) mol/L) or VCR (0, 10(-7), 10(-6), 10(-5), 10(-4) mol/L), ADR (0, 10(-6), 10(-5), 10(-4), 10(-3) mol/L), or ACT-D (0, 5 X 10(-8), 1 X 10(-7), 5 X 10(-7), and 10(-6) mol/L) . The cells were then tested for bacterial killing against Staphylococcus aureus, chemotaxis activity stimulated by Escherichia coli endotoxin, N-formyl-methionyl-leucyl-phenylalanine (FMLP)-stimulated aggregation, and cytochalasin B (Cyto B)/FMLP-stimulated superoxide production and enzyme degranulation . High concentration of CTX, an alkylating agent, showed a significant depression of PMN superoxide production, (124 +/- 13 v 161 +/- 15 nmol/10(7) cells, 5 minutes, P less than or equal to .025) . ADR, an intercalating agent and membrane inhibitor, showed a significant depression of PMN degranulation and lysozyme release at 10(-4) and 10(-3) mol/L (15.3% +/- 1.7% v 24% +/- 7%, P less than .01; and 15.0% +/- 2.5% v 24% +/- 7%, P less than or equal to .025) . VCR, a microtubule inhibitor, showed a significant depression of PMN aggregation at 10(-6), 10(-5), and 10(-4) mol/L (P less than .05), lysozyme release at 10(-4) mol/L (P less than .004), and beta-glucuronidase release at 10(-4) mol/L (P less than .004) . In addition, chemotaxis was inhibited by VCR in a dose-dependent manner at all concentrations (10(-7) mol/L, P less than .02; 10(-6) mol/L, P less than .007; 10(-5) mol/L, P less than .006, and 10(-4) mol/L, P less than .003) . ACT-D showed no significant effect on the PMN functions tested . These studies conclude that chemotherapeutic agents have modulating in vitro effects on PMN function . Further in vivo studies are therefore needed to assess PMN abnormalities in patients receiving cancer chemotherapy to determine their role in infectious complications.

J Bacteriol, 1986 May, 166(2), 574 - 80
Cloning and expression of the exfoliative toxin B gene from Staphylococcus aureus; Jackson MP et al.; Exfoliative toxin type B is produced by bacteriophage group II strains of Staphylococcus aureus and is a causative agent of staphylococcal scalded-skin syndrome . In addition to exfoliative toxin B, most isolates also produce a bacteriocin and are immune to the action of the bacteriocin . These phenotypes, as well as resistance to cadmium, were lost after elimination of a 37.5-kilobase plasmid, pRW001, from S . aureus UT0007 . Transduction and transformation showed that pRW001 carries the structural genes for four phenotypic characteristics of S . aureus UT0007: (i) exfoliative toxin B production, (ii) bacteriocin production, (iii) bacteriocin immunity, and (iv) resistance to Cd(NO3)2 . The exfoliative toxin B structural gene (etb), which is located on a 1.7-kilobase HindIII fragment of pRW001, was cloned in the plasmid pDH5060 and transformed into phage group III S . aureus RN4220 . Transformant clones produced extracellular exfoliative toxin B that was biologically active in the neonatal mouse assay . In the Escherichia coli genetic background, the exfoliative toxin B gene was expressed only after being cloned into the positive selection-expression vector pSCC31 . The structural gene for cadmium resistance was also isolated on an HindIII fragment of pRW001 cloned in pDH5060 . The loci for the exfoliative toxin B gene and the cadmium resistance gene(s) were identified on a restriction map of plasmid pRW001.

J Bacteriol, 1986 May, 166(2), 385 - 91
Lysogenic conversion of staphylococcal lipase is caused by insertion of the bacteriophage L54a genome into the lipase structural gene; Lee CY et al.; Staphylococcus aureus PS54 manifests no lipase (geh) activity . This is due to the insertion of bacteriophage L54a DNA into the geh structural gene . The nucleotide sequence of this 2,968-base-pair DNA fragment was determined . Lipase deduced from the nucleotide sequence is a polypeptide of 690 amino acids which extends from nucleotide 706 to 2776.

J Immunol, 1986 May 1, 136(9), 3447 - 54
Metabolic comparison between basophils and other leukocytes from human blood; de Boer M et al.; Basophilic granulocytes were purified from the blood of normal individuals by successive isopyknic centrifugation and elutriation centrifugation . Starting with the leukocyte-rich fraction of 500 ml of blood, we recovered 31 to 80% (mean 51%, n = 20) of the basophils in 45 to 87% purity (mean 69%, n = 23) . The contaminating cells were mainly lymphocytes . The basophils were greater than 98% vital (exclusion of ethidium bromide and hydrolysis of fluorescein diacetate) . The histamine content of the basophils was 1.1 to 2 pg/cell (mean 1.6 pg/cell, n = 22) . With anti-IgE, 30 to 50% of the histamine was released; with phorbol myristic acetate (PMA) or the calcium ionophore A23187, 70 to 100% of the histamine was released . Serum-opsonized zymosan (STZ) did not induce histamine release . Reactions with monoclonal antibodies revealed that the basophils expressed the C3bi receptor (CR3) and the leukocyte function-associated antigen 1 (LFA1), but not the gp 150,95 antigen, the C3b receptor (CR1), or the low avidity Fc gamma receptor . Basophils carry class I but not class II HLA antigens . During incubation of the basophils with serum-opsonized Staphylococcus aureus or Escherichia coli, these bacteria were neither phagocytized nor killed . STZ, PMA, A23187, or anti-IgE did not initiate an "oxidative burst" in the basophils . This was tested with oxygen consumption, cytochrome c reduction, NBT reduction, chemiluminescence, and release of hydrogen peroxide . Moreover, we did not detect cytochrome b558, superoxide dismutase, catalase, or peroxidase in the basophils . Of the typical granule-associated enzymes lysozyme, Vitamin B12-binding protein, and beta-glucuronidase, only beta-glucuronidase was present in the basophils in detectable amounts . This enzyme was released, together with histamine, on incubation of the cells with PMA, A23187, or anti-IgE, but not with STZ . We conclude that basophils from normal human blood are not phagocytes and are probably not involved in the oxidative defense of the host against foreign antigens.

Tohoku J Exp Med, 1986 May, 149(1), 95 - 102
Opsonic activity of plasma fibronectin for Staphylococcus aureus by human alveolar macrophages: inefficacy of trypsin-sensitive staphylococcal fibronectin receptor; Oishi K et al.; The binding of 125I-fibronectin (FN) to some bacteria and the phagocytic activity of human alveolar macrophages (AM) by a culture method using 3H-labelled bacteria were examined . FN-binding of Staphylococcus aureus (S . aureus) was higher than that of gram-negative rods and was reduced markedly by trypsin-treatment . FN enhanced the uptake of S . aureus by AM in dose dependent manner . To determine whether FN-binding to S . aureus mediates the promotion of staphylococcal phagocytosis, we compared the uptake of non-treated S . aureus with that of trypsin-treated bacteria by AM . No significant difference was found between the uptake of these bacteria and no effect of FN-binding to S . aureus on the phagocytic activity . It is concluded that FN plays an essential role as a non-immune opsonin for S . aureus in the human lung which is found to be abundant in FN, and that FN-binding to S . aureus is probably not related to this effect.

Infection, 1986 May-Jun, 14(3), 136 - 8
Treatment of experimental chronic osteomyelitis due to Staphylococcus aureus with teicoplanin; Norden CW et al.; Teicoplanin was used alone in the treatment of experimental osteomyelitis due to Staphylococcus aureus in rabbits . Treatment with 60 mg of teicoplanin/kg of body weight twice a day for 28 days failed to sterilize any infected rabbit bones . A possible explanation for the failure of teicoplanin may be that its in vitro activity against the infecting strain of S . aureus was substantially less under anaerobic conditions (at partial pressures of oxygen analogous to those in osteomyelitic rabbit bones) than under aerobic conditions.

J Vasc Surg, 1986 May, 3(5), 732 - 40
Bacterial adherence to vascular prostheses . A determinant of graft infectivity; Schmitt DD et al.; An in vitro model was developed to quantitatively measure bacterial adherence to the surface of prosthetic vascular graft material . Four strains of bacteria (Staphylococcus aureus, nonmucin-producing S . epidermidis {SP-2}, mucin-producing S . epidermidis {RP-12}, and Escherichia coli) were used to inoculate expanded polytetrafluoroethylene (ePTFE), woven Dacron, and velour knitted Dacron graft material . After graft specimens were incubated in a 10(7) suspension of bacteria, they were washed to remove nonadherent organisms and ultrasonically oscillated to dislodge adherent organisms . Quantitative culture of the sonication effluent was used to calculate bacterial adherence, expressed as the number of colony-forming units found in each square centimeter of graft material per 10(7) inoculum . All bacterial strains had a greater affinity to velour knitted Dacron graft than to ePTFE (p less than 0.025) . E . coli and S . aureus adhered to velour knitted Dacron in greater numbers than to woven Dacron (p less than 0.04) . The production of extracellular polysaccharide (mucin) by the RP-12 strain significantly increased adherence to both EPTFE and Dacron grafts compared with the other three bacterial strains tested (p less than 0.04) . Although E . coli was less adherent to ePTFE than nonmucin-producing staphylococcal strains (S . aureus and SP-2), no difference in adherence to knitted or woven Dacron graft material was demonstrated . The differential adherence of bacteria to prosthetic vascular grafts pays an important role in the pathogenesis of graft sepsis and determines relative graft infectivity . The in vitro model developed is well suited for further study of the mechanisms by which bacteria adhere to and colonize vascular grafts.

Hand Clin, 1986 May, 2(2), 271 - 90
Silicone implants; Kleinert JM et al.; Infection has not been considered in this article with each individual implant . The incidence is low indeed . In the only publication concerned primarily with the topic of infection following silicone implant surgery, Millender et al . reviewed 2105 implants of varying kinds . There were ten infections, seven of which were with Staphylococcus aureus . The onset was remarkably late--17 days after surgery on average . In seven cases the implant had to be removed and the eventual result was good, being likened to that obtained after an excisional arthroplasty . Reviewing the complications that occur with the various implants, it becomes evident that there are three primary concerns--fracture, subluxation, and synovitis . Fracture occurs primarily in the wrist and the metacarpophalangeal implants . The incidence of fracture in the wrist implant is 8.6, 9.4, and 19.8 per cent, giving an average of the means of 12.6 per cent . In the metacarpophalangeal joint, the incidence with the Swanson design is variously 1.9, 26.2 and 21 per cent, the average of the means being 16.4 per cent . The Niebauer design is reported as having a fracture rate of 29.7 and 38 per cent, for an average of the means of 33.9 per cent . The somewhat lower incidence of fracture of the wrist implant is offset by the fact that, in contrast to the situation with the smaller joint, the fracture is almost always symptomatic, requiring treatment . Largely for this reason, silicone wrist arthroplasty is limited mainly to the rheumatoid patient, being little used for post-traumatic arthritis . Subluxation of implants occurs mainly with the carpal replacements . The incidence in independent reports are 56.5 and 50 per cent, for an average of the means of 53.3 per cent with the scaphoid; 20, 20, and 50 per cent for an average of the means of 30 per cent with the lunate; and 5.3, 10, 11.2, 29, and 32 per cent for an average of the means of 17.5 per cent with the trapezium . In the case of the trapezium, excision of a portion of the trapezoid, supplemented where necessary by ligament reconstruction to support the first metacarpal, appears to give the hope of lowering the incidence of subluxation to an acceptable level . With the lunate, preservation of an anterior shell may give satisfactory results but judgment should await longer term studies of larger groups . The scaphoid implant gives most cause for concern, both because the incidence is high and because the solutions offered have either failed or are too recent to judge and perhaps too radical to accept.(ABSTRACT TRUNCATED AT 400 WORDS)

J Hosp Infect, 1986 May, 7(3), 236 - 43
Disinfectant induced changes in the antibiotic sensitivity and phage typing pattern in Staphylococcus aureus; Sivaji Y et al.; Serial passage through disinfectants at sub-minimal inhibitory concentrations induced antibiotic resistances in 30.5% of the derived phenotypic variants of five strains of Staphylococcus aureus . A proportion of these phenotypic variants showed changes in their phage typing pattern by becoming susceptible to an increased number of phages, and a change from phage non-typable to typable was also observed . A loss of temperate phages was noted in some variants when subjected to reverse phage typing.

J Biol Chem, 1986 Apr 25, 261(12), 5651 - 7
Peptide mapping analysis of the avian progesterone receptor; Puri RK et al.; Progesterone receptor from the chicken oviduct has been shown to exist as two 8 S forms (I and II) . Form I contains a protein of Mr = 75,000 and form II contains a protein of Mr = 110,000 . In addition to these hormone-binding proteins, both receptor forms contain a protein with Mr = 90,000 that does not bind steroid . To investigate the possibility that these proteins are structurally related, they were isolated by preparative sodium dodecyl sulfate gel electrophoresis and subjected to peptide mapping analyses after digestion with Staphylococcus aureus V-8 protease, papain, or alpha-chymotrypsin . Receptor proteins labeled with {32P}orthophosphate in tissue minces were also subjected to peptide mapping analysis . The electrophoretic patterns of peptide fragments of the 90-kDa protein from receptor forms I and II were identical but were different from the peptide patterns obtained from the 75- and 110-kDa proteins which generated similar peptide patterns, indicating that these are structurally related . However, some differences were evident, indicating that these latter two proteins are not identical substrates for proteases . A one-dimensional comparison of the phosphopeptide patterns from the 75- and 110-kDa proteins also showed them to be similar, but not identical . Two-dimensional maps of phosphopeptides generated from the 75- and 110-kDa protein after complete tryptic digestion revealed multiple sites of phosphorylation which were identical except for one phosphopeptide that was unique to the 110-kDa protein . These results show the two progesterone-binding proteins to be very similar in structure, but to differ considerably from the 90-kDa protein.

J Biol Chem, 1986 Apr 15, 261(11), 5034 - 40
Tyrosine sulfation in precursors of collagen V; Fessler LI et al.; Radioactive labeling of p-collagens V, collagens V, and, to a small extent, of procollagen V occurred when {35S}sulfate was incubated with tendons or primary tendon cell cultures, or blood vessels and crops of 17- to 19-day-old chick embryos, or with lung slices from neonatal rats . Most or all of this label is in the form of 1 or more sulfated tyrosine residues/chain of p alpha 1(V), alpha 1(V), p alpha 1'(V), alpha 1'(V), p alpha 2(V), and alpha 2(V), and it remains attached through purification by dialysis, ammonium sulfate precipitation, CsCl-GdnCl2 equilibrium buoyant density and velocity sedimentations, ion-exchange chromatography, and sodium dodecyl sulfate gel electrophoresis . Radioactive tyrosine sulfate was identified in alkaline hydrolysates of these collagen V chains, after labeling the tissues with either {35S}sulfate or {3H}tyrosine, by electrophoretic and chromatographic comigration with a tyrosine sulfate standard . Tunicamycin A1, which inhibits the attachment of N-linked complex carbohydrate, did not interfere with the sulfation process . The tyrosine sulfate is located in a noncollagenous domain, which is probably adjacent to the amino end of the collagen helix, and is retained throughout the physiological proteolytic processing of procollagens V . After digestion with Staphylococcus aureus V8 protease, 35S-labeled p alpha 1(V) and alpha 1(V) chains gave the same map of labeled peptides, and this differed from the map given by p alpha 1'(V) and alpha 1'(V) chains . Little sulfation of p alpha 2(V) and alpha 2(V) chains occurs . The implications of these observations for the structure and properties of procollagens V and their derivatives are considered.

J Biol Chem, 1986 Apr 5, 261(10), 4594 - 9
Isolation and characterization of a 37,000-dalton protein associated with the erythrocyte membrane; Lin T et al.; We have purified a 37,000-dalton polypeptide (p37) from the red cell membrane that was found in previous studies to undergo a lineage-specific alteration in its membrane association . Our data suggest that p37 associates with the red cell membrane through electrostatic interactions that are resistant to 0.5 M NaCl or 10 mM EDTA . Conditions found to elute p37 from red cell ghosts include H2O at pH 12, 0.1 N NaOH + 1 mM ethanol and 1.0% Triton X-100 . p37 was purified substantially from ghosts by Triton X-100 solubilization followed by sequential DEAE-Sephadex and CM-Sephadex chromatography . When p37 was analyzed by two-dimensional gel electrophoresis, a family of isoelectric focusing variants was detected ranging in pI from 7.0 to 7.8 . All of the isoelectric focusing variants showed homology to one another when compared serologically with anti-p37 antibodies or by limited peptide mapping using Staphylococcus aureus V8 protease . The isoelectric focusing variants appear to represent distinct, yet related polypeptides rather than degrees of post-translational modifications to a single species, inasmuch as all of the variants are present in anti-p37 immunoprecipitates prepared from in vitro translations programmed with p37 mRNA.

J Biol Chem, 1986 Apr 5, 261(10), 4749 - 57
Phosphorylation of rhodopsin by protein kinase C in vitro; Kelleher DJ et al.; Calium/phospholipid-dependent protein kinase (protein kinase C) was purified from bovine retinae rod outer segments (ROS) . In the presence of 0.1-2 microM calcium protein kinase C binds tightly to ROS and phosphorylates rhodopsin in the absence or presence of illumination . This property of protein kinase C contrasts with that of rhodopsin kinase, which in vitro phosphorylates only bleached rhodopsin . Peptide maps of rhodopsin phosphorylated by protein kinase C or rhodopsin kinase were compared using limited Staphylococcus aureus V8 protease digestion or complete tryptic digestion . Phosphorylation sites map to serine and threonine residues on the cytoplasmic carboxylterminal domain of rhodopsin for both kinases . The functional consequence of protein kinase C phosphorylation of rhodopsin was a reduced ability to stimulate the light-dependent rhodopsin activation of {35S}guanosine 5'-O-(thiotriphosphate) binding to transducin, the GTP-binding regulatory protein present in ROS . Properties of the calcium-stimulated interaction of protein kinase C with membranes and in vitro phosphorylation of intrinsic proteins are discussed based upon the findings.

J Bacteriol, 1986 Apr, 166(1), 29 - 33
Nucleotide sequence of the enterotoxin B gene from Staphylococcus aureus; Jones CL et al.; The complete nucleotide sequence of the enterotoxin B gene from Staphylococcus aureus, as well as the 5' and 3' flanking regions, was determined . Starting from an ATG initiator codon, an open reading frame encoded the enterotoxin B precursor that consisted of 266 amino acids (Mr, 31,400) . The 5' terminal portion of the gene encodes a signal peptide 27 amino acids long . The deduced amino acid sequence matched, with a few exceptions, the published amino acid sequence of enterotoxin B . The structural gene was flanked on the 5' side by a promoter-like sequence and on the 3' side by a palindromic structure followed by a thymine-rich region that resembled a transcription termination signal . Downstream from the entB gene were two overlapping open reading frames corresponding to 134 and 185 amino acids in the opposite orientation . The signal sequence of the enterotoxin B precursor resembled that of other secreted proteins found in other bacteria.

Infect Immun, 1986 Apr, 52(1), 331 - 3
Hormonal influence on experimental infections by a toxic shock strain of Staphylococcus aureus; Best GK et al.; Subcutaneous infection chambers in rabbits were infected with a strain of Staphylococcus aureus isolated from a patient with toxic shock syndrome . Estrogens (mestranol and 17-beta-estradiol) protected male rabbits and prolonged survival . Neither androgens (testosterone and dihydrotestosterone) nor progesterone affected the susceptibility of intact or ovarihysterectomized female rabbits.

Pathology, 1986 Apr, 18(2), 227 - 33
Acute hematogenous staphylococcal osteomyelitis: the effects of surgical drilling and curettage in an animal model; Emslie KR et al.; Surgical drilling and curettage of metaphyseal medullary bone were performed in chickens 4, 8 and 15 d after bacterial inoculation with Staphylococcus aureus producing osteomyelitis . Intramedullary drilling and curettage resulted in extensive vascular damage and necrosis of medullary bone but failed to remove all viable bacteria . An effective continuing drainage channel for the pus was not established due to the formation of blood clot within the drill hole . Surgery performed before the formation of a sequestrum resulted in further spread of bacteria within the medullary cavity and the formation of a larger sequestrum than that commonly observed in the bones of infected, untreated chickens . Surgery performed after the formation of a sequestrum destroyed the abscess wall thus disrupting a natural defence mechanism of the host . Whilst a direct comparison of surgical drilling in human osteomyelitis and in this avian model cannot be made, the results suggest that production of an artificial drill hole across the cortex and curettage of the bone in human osteomyelitis would be destructive and may not be warranted.

Scott Med J, 1986 Apr, 31(2), 85 - 9
Peritonitis in continuous ambulatory peritoneal dialysis; Smith WG et al.; The main complication of continuous ambulatory peritoneal dialysis (CAPD) is peritonitis . This paper describes our experience in the diagnosis and management of this complication in 66 patients during the three years to October 1982 . The overall incidence of peritonitis was one episode every 6.75 patient months . Staphylococcus albus and Staphylococcus aureus together accounted for 46 per cent of the episodes, and 24 per cent were culture negative . Catheter exit site infections due to Staphylococcus aureus were common and they may have predisposed to peritonitis with gram -ve organisms as well as to staphylococcal peritonitis . Antimicrobial therapy was effective in 60 per cent of peritonitis episodes . The culture negative episodes usually responded to treatment while those due to fungi, though uncommon, did not . Twenty-nine per cent of these CAPD patients were transferred to haemodialysis because of peritonitis which failed to respond to treatment or which recurred repeatedly.

Gan To Kagaku Ryoho, 1986 Apr, 13(4 Pt 2), 1201 - 7
{Experimental study on the clonogenic assay--in vitro and in vivo chemosensitivity tests of human tumor xenografts serially transplanted into nude mice}; Kubota T; In order to determine the optimal conditions of the clonogenic assay for predicting antitumor activity in vivo, experimental chemosensitivity tests in vitro and in vivo were performed using a human gastric carcinoma strain, H-111 serially transplanted into nude mice . According to the method of Salmon and Hamburger, 5-fluorouracil (5-FU) was added to dissociated tumor cells at concentrations of 0.1, 1 and 10 micrograms/ml continuously for 2 weeks in the upper layer of a double agar preparation . The transplanted H-111 tumors in nude mice were treated with 30, 60 and 120 mg of 5-FU per kg . The antitumor activities were evaluated by the T/C ratio of the colonies in vitro and the T/C ratio of tumor weights in vivo . Pharmacokinetic analysis of 5-FU was conducted by bioassay with Staphylococcus aureus in the agar and in the serum of mice . When areas under the curves in vitro and in vivo were supposed to be equivalent, the antitumor activities of 5-FU in vitro and in vivo could be correlated quantitatively in terms of T/C ratios . The drug concentration in the clonogenic assay for predicting the effect of the MTD of 5-FU in vivo (50 mg/kg X 3) was calculated to be 1 microgram/ml for 2 weeks.

Virologie, 1986 Apr-Jun, 37(2), 89 - 93
Immunochemical aspects of cerebrospinal fluid immunoglobulins in patients with subacute sclerosing panencephalitis; Cernescu C et al.; Unconcentrated cerebrospinal fluid (CSF) samples from 50 patients with subacute sclerosing panencephalitis (SSPE) were investigated by rocket immunoelectrophoresis (RIE) against anti-human IgG sera . Two patterns of immune precipitates were observed: precipitates with exclusively cathodal and precipitates with bipolar extension . Hemagglutination-inhibition tests against measles virus antigen were performed after absorption of the CSF samples with Protein A-containing Staphylococcus aureus and treatment with 2-mercaptoethanol . The results point to different pathological significances of the precipitation line anomalies revealed by RIE, that might be relevant for some of the patients investigated.

Mol Immunol, 1986 Apr, 23(4), 377 - 84
Modulation of IgG effector functions by a monovalent fragment of staphylococcal protein A; Ghetie V et al.; The monovalent V-1 fragment of protein A (fSpA) with a mol . wt of 13,000 obtained from an u.v . mutant of Staphylococcus aureus Cowan I strain was proved to be able to modulate significantly some of the effector functions of IgG, such as complement fixation, catabolism, attachment to Fc receptors and antibody-dependent cell-mediated cytotoxicity . Moreover fSpA-like protein A obtained from the A676 strain is mitogenic and enhances NK activity of human peripheral lymphocytes . The efficiency of fSpA was found to be lower than that of protein A with regard to its ability to inhibit complement fixation, EA rosette formation and antibody-dependent cell-mediated cytotoxicity . Both protein A and fSpA had the same efficiency in activation of the complement system after reaction with human or guinea pig IgG, and in increasing the IgG catabolism . Unlike fSpA the monovalent B fragment of protein A (with mol . wt of 7000) was not able to inhibit EA rosette formation and antibody-dependent cell-mediated cytotoxicity . The results recommend fSpA, substituting for protein A, as a molecular probe for the investigation of IgG antibody and lymphocyte effector functions.

Microbiologica, 1986 Apr, 9(2), 209 - 14
Chemiluminescence, phagocytosis, chemotaxis and killing activity of human leukocytes exposed to Clindamycin; Scevola D et al.; We measured the chemiluminescence (CL) of human leukocytes (PMNs) exposed to different concentrations of Clindamycin (1; 5; 1 mcg/ml) using a standardized luminolamplified reagent-instrument methodology (Auto Picolite 6500 Luminometer and ZAP/Picolite Kits, Packard Instruments Co . Drowners Grove, IL . USA) . Cells obtained from healthy donors were also tested for chemotaxis with Boyden chambers, for phagocytosis and for killing activity against Staphylococcus aureus by the agar-medium culture plate technique . Clindamycin does not induce significant variations of the CL response in whole blood, or changes in phagocytosis and killing activity . On the contrary, concentrations of drug corresponding to therapeutically obtainable levels significantly increase light emission by isolated cells . A concentration effect was seen on leukotaxis, that was increased, but not significantly, at 1 and 5 mcg/ml and decreased (P less than 0.01) at 10 mcg/ml . CL assay is a simple, sensitive, reproducible technique to assess the PMNs functions during antibiotic therapy.

J Antimicrob Chemother, 1986 Apr, 17(4), 409 - 13
Beta-lactam antibiotics increase the frequency of plasmid transfer in Staphylococcus aureus; Barr V et al.; The transfer of a plasmid specifying tetracycline resistance between different derivatives of Staphylococcus aureus by phage mediated conjugation was enhanced 100- to 1000-fold by exposure of the culture to subinhibitory concentrations of beta-lactam agents . A variety of other antibiotics, including vancomycin and teicoplanin, had no such effect . The enhanced frequency of transfer was probably due to the formation of large bacterial aggregates.

Appl Environ Microbiol, 1986 Apr, 51(4), 699 - 702
Evaluation of culture media for recovery of Staphylococcus aureus from swimming pools; Alico RK et al.; Several selective media were evaluated for the primary isolation and enumeration of Staphylococcus aureus from halogenated indoor swimming pool waters . Standard plate counts of the viable population and total coliform densities were also determined to ascertain their value as indicator systems . All studies were done with membrane filters . The most selective, accurate, and reliable medium was Vogel-Johnson (VJ) medium supplemented with 0.5% pyruvate . This medium recovered two times more typical colonies than VJ medium alone, and subsequent identification of these well-defined black colonies proved that approximately 80% were S . aureus . The S . aureus recoveries correlated well with halogen levels and bather density use also . In contrast, VJ medium alone was 60% selective for S . aureus, and VJ medium supplemented with catalase did not increase either the percent recovery or the selectivity over that of VJ medium alone . Standard plate counts did not correlate with halogen levels, bather density, or total viable colonies . Coliforms were rarely recovered from indoor pool waters and were not considered to be useful indicators of water quality.

Antimicrob Agents Chemother, 1986 Apr, 29(4), 611 - 3
Serum bactericidal activity of rifampin in combination with other antimicrobial agents against Staphylococcus aureus; Hackbarth CJ et al.; Because rifampin-resistant strains of Staphylococcus aureus emerge during monotherapy with this drug, a search was made for potentially useful companion drugs . Bactericidal titers of spiked serum were determined, and time kill studies were performed for 10 strains of methicillin-susceptible S . aureus . We tested rifampin in combination with nafcillin, vancomycin, clindamycin, pefloxacin, ciprofloxacin, trimethoprim, teicoplanin, or erythromycin . The bactericidal activity of nafcillin, vancomycin, and teicoplanin was significantly reduced (P less than 0.05) when rifampin was added to the drug regimen . In contrast, the addition of rifampin to clindamycin or erythromycin significantly increased bactericidal activity as measured by both bactericidal titers in serum and 6-h killing rates (P less than 0.02) . Bactericidal activity in serum was also increased by the addition of rifampin to trimethoprim, but rifampin-resistant strains emerged with this combination.

Neurosurgery, 1986 Apr, 18(4), 461 - 4
Isolated C-2 osteomyelitis of hematogenous origin: case report and literature review; Venger BH et al.; A case of hematogenous Staphylococcus aureus osteomyelitis isolated to the 2nd cervical vertebra is presented . To our knowledge, this is the second case to be reported of hematogenous infection involving only the body and odontoid process of the axis . The first report was published prior to the advent of computed tomography, bone scans, and halo orthosis . The pathophysiology of blood-borne atlantoaxial infection is described, as well as methods of diagnostic evaluation and therapeutic recommendations . The use of computed tomography to define the extent of the lesion is illustrated.

Antimicrob Agents Chemother, 1986 Apr, 29(4), 663 - 9
Resistance to mercury and to cadmium in chromosomally resistant Staphylococcus aureus; Witte W et al.; Apparently chromosomally located mercury resistance determinants in five methicillin-resistant Staphylococcus aureus strains of different geographical origin were structurally homologous to plasmid-located mercury resistance determinants in S . aureus . These were all located on a 6.3-kilobase (kb) Bg/II fragment, as evident from Southern hybridization experiments with the 6.3-kb Bg/II fragment of plasmid pI258 as the probe . These methicillin-resistant S . aureus strains exhibited similar phage susceptibility patterns and biochemical reactions . They differed, however, in the DNA location of the mercury resistance determinants, as evidenced by neighboring cleavage sites for restriction endonucleases EcoRI, HindIII, and PstI . In an environmental (nonhospital) strain in which mercury resistance was also apparently chromosomally conferred, these determinants were also homologous to pI258 DNA, but they were located on a 6.6-kb Bg/II fragment . Cadmium resistance determinants in the five methicillin-resistant S . aureus strains and the environmental S . aureus strain were not similar to the known plasmid-located determinants cadA and cadB . Cd2+ resistance was based on an efflux mechanism for Cd2+ . However, no parallel resistance to zinc was conferred . The 3.2-kb XbaI-Bg/II fragment obtained from plasmid pI258 and used as a cadA-specific probe did not hybridize to total DNA digests of the strains with apparently chromosomally determined cadmium resistance.

J Hyg (Lond), 1986 Apr, 96(2), 217 - 23
Bacteriological characters of strains of Staphylococcus aureus submitted to a reference laboratory related to methicillin resistance; Marples RR et al.; Methicillin-resistant Staphylococcus aureus (MRSA) strains present an increasing clinical problem . Analysis of 2679 strains submitted to a reference laboratory in the first quarter of 1983 and 3050 strains submitted in summer 1984 showed 479 and 593 multi-resistant strains . The proportion of methicillin-resistant strains classified as epidemic rose from 5.9 to 10.2% . Other methicillin-resistant strains continued to occur but other methicillin-sensitive multi-resistant strains appeared to fall . A strain with defined characters could be recognized in the Thames regions.

J Clin Pathol, 1986 Apr, 39(4), 371 - 5
Coagulase production by strains of Staphylococcus aureus of differing resistance characters: a comparison of two traditional methods with a latex agglutination system detecting both clumping factor and protein A; Dickson JI et al.; Five groups of strains of Staphylococcus aureus (54 in total) were tested by slide and tube coagulase methods with rabbit and human plasma, and the results were compared with a latex test for both clumping factor and Protein A (Staphaurex, Wellcome Foundation) . The five groups comprised: epidemic methicillin resistant S aureus (group 1); other methicillin resistant S aureus (group 2); other resistant S aureus (group 3); other S aureus (group 4); and a group of reference strains, not all true S aureus (group 5) . Groups 1, 3, and 4 gave consistently strong positive results with the tube test and the latex test and less strong positive results with the slide test . Group 2 strains sometimes gave weak or negative results in slide and latex tests, but tube tests with both types of plasma were strongly positive . Only within group 5 strains were negative results in the tube test found . Group 1 strains showed no diminution in expression of free coagulase or of clumping factor . The latex test was more sensitive than the slide test but less sensitive than the tube test . Doubtful or negative slide test or latex test results, particularly with strains resistant to methicillin, should be checked by a tube coagulase test.

Pathol Biol (Paris), 1986 Apr, 34(4), 245 - 8
{In vitro activity of the combination ticarcillin-clavulanic acid on bacterial isolates in surgery}; Branger C et al.; The in vitro activity of ticarcillin in combination with clavulanic acid was tested, by disc diffusion, against 1,380 clinical bacterial isolates and was compared with that of ticarcillin alone . 83.8% of the isolates were susceptible to ticarcillin + clavulanic acid, whereas 56.6% were susceptible to ticarcillin alone . Minimal inhibitory concentrations (MIC) of ticarcillin in the presence of 4 micrograms/ml of clavulanic acid were determined against 157 ticarcillin resistant (MIC greater than 128 micrograms/ml) but ticarcillin + clavulanic acid susceptible strains of Gram negative bacilli and against 20 strains of beta-lactamase producing Staphylococcus aureus . With the addition of clavulanic acid, MICs of ticarcillin were respectively less than or equal to 16 micrograms/ml and less than or equal to 64 micrograms/ml for 50 and 90% of the Gram negative bacilli . All the Staphylococcus aureus were inhibited by concentrations of ticarcillin less than or equal to 1 microgram/ml.

Microb Pathog, 1986 Apr, 1(2), 125 - 38
Inactivation of the alpha-haemolysin gene of Staphylococcus aureus 8325-4 by site-directed mutagenesis and studies on the expression of its haemolysins; O'Reilly M et al.; S . aureus strain 8325-4 was shown to produce alpha-, beta-, delta- and gamma-haemolysins by haemolytic assays and immunoblotting . Hybridization experiments indicated that a single copy of the alpha-haemolysin gene (hla) resides in the chromosome . Site-directed mutagenesis was used to inactivate the hla gene . This gene, which had previously been cloned in E . coli, was inactivated in vitro by inserting a fragment carrying an erythromycin resistance marker . Shuttle plasmids were constructed and transformed into 8325-4 and non-haemolytic recombinants enriched by a plasmid incompatibility technique . A previously isolated Tn551 insertion defective in alpha-haemolysin was not located in hla . It had pleiotropic defects in expression of alpha-, beta- and delta-haemolysins . Expression of alpha-haemolysin from a plasmid-located hla gene was very low . In contrast, hla-erm mutants were deficient only in alpha-haemolysin and allowed high level expression of the plasmid-borne hla gene . The Tn551 insertion is probably located in a gene encoding a positive regulatory element required for expression of several exoproteins . An hla-erm mutant was less virulent than the otherwise isogenic 8325-4 hla+ strain in a mouse peritonitis model, confirming that alpha-haemolysin is an important virulence factor.

J Immunol, 1986 Apr 1, 136(7), 2375 - 81
Antibodies reactive with class II antigens encoded for by the major histocompatibility complex inhibit human B cell activation; Clement LT et al.; Although class II antigens encoded by genes in the major histocompatibility complex (MHC) are important as recognition structures for immunoregulatory cell interactions, the precise functional role of these molecules in the biological responses of B lymphocytes is unknown . In the studies described here, we have examined the effects of six monoclonal antibodies reactive with human class II MHC antigens on B cell activation and proliferation . Peripheral blood IgM+ B cells purified by fluorescence-activated cell sorter (FACS) techniques were stimulated with anti-mu antibodies, protein A-bearing Staphylococcus aureus (SAC), or in T cell-dependent activation cultures . The B cell proliferative responses induced by these stimuli were inhibited 68 to 90% by low concentrations (1 to 5 micrograms/ml) of antibodies reactive with class II MHC antigens . Antibodies specific for DR and DQ antigens were both effective inhibitors of B cell proliferation . This inhibition was not due to the binding of antibody to B cell Fc-IgG receptors, because IgM and IgG anti-class II antibodies were equally potent as inhibitors . When responses of B cells fractionated on the basis of cell size by forward angle light scatter were analyzed, anti-DR and anti-DQ antibodies inhibited the proliferation of small, resting IgM+ cells induced by T-independent as well as T-dependent stimuli . Activation-dependent increases in B cell size and RNA synthesis were similarly inhibited . In contrast, the responses of large B cells (that had been preactivated in vivo) to T cell-derived B cell growth factors were not affected by anti-class II antibodies . These data suggest that class II MHC molecules do not serve merely as cellular interaction structures but also directly participate in early events of the B cell activation cascade that precede cell enlargement or increased RNA synthesis . After activation and expression of receptors for growth factors, however, B cell class II MHC antigens no longer mediate signals required for mitogenesis.

Gan To Kagaku Ryoho, 1986 Apr, 13(4 Pt 2), 1314 - 21
{Comparative studies of human natural IFN-alpha, IFN-beta and IFN-gamma with regard to their effects on in vitro immune responses}; Aoki N et al.; Interferon (IFN) is known to affect a variety of immune responses apart from its well-established antiviral and antineoplastic effects . Three human IFN species: IFN-alpha, IFN-beta and IFN-gamma are now clinically available as a result of recent rapid improvements in IFN technology . In view of only the scanty data presently available concerning comparative studies among IFNs with regard to their effects on immune regulation, this report deals comparatively with natural IFN-alpha, IFN-beta and IFN-gamma (all products of Green Cross) with respect to their effects on natural killer cell (NK) activity, chemiluminescence (CL) of fractionated NK cells, lymphoproliferative response induced by PHA, PWM and Staphylococcus aureus COWAN-1 (SAC), and IL-2 production under stimulation with PHA (1:1000) . Comparison among IFNs for these immune responses was performed with the same level of antiviral activity (IU) . NK activity in the peripheral blood was enhanced in the presence of IFN of all species: to the same extent with IFN-alpha and IFN-beta, but significantly less for IFN-gamma as compared with the former . CL of NK cells to IFN was again similar in extent for IFN-alpha and -beta, but less for IFN-gamma . Inhibition of PHA and PWM blastogenesis was similar for IFN-alpha and -beta but less for IFN-gamma as shown previously . SAC blastogenesis of both peripheral lymphocytes and tonsillar B cells was enhanced in the presence of IFN-gamma but not in the presence of IFN-alpha or -beta . IL-2 production of peripheral lymphocytes by PHA (1:1000) was dose-relatedly enhanced in the presence of IFN-gamma but not in the presence of IFN-gamma or -beta.

Vet Med (Praha), 1986 Apr, 31(4), 233 - 44
{Physical, chemical and biological study of dust from large-scale pig farms}; Raszyk J; Dust deposition in 16 halls of two large pig-fattening farms with dry or wet feeding systems was analyzed . In the halls with wet feeding the samples contained maximally 28 dust particles up to 10 micron and 17 particles up to 5 micron per cm3 of air, in the halls with dry feeding 220 particles smaller than 10 micron and 205 particles smaller than 5 micron per cm3 of air . The total amino acid content in the dust deposition was 17.440 +/- 1.820 g per 100 g of sample and the content of nitrogen compounds (N X X 6.25, %), was 24.170 +/- 2.910 . The contents of chemical elements were as follows (mg per kg): zinc 448 +/- 151; manganese 109.9 +/- 49.5; copper 40.5 +/- 12.1; lead 4.77 +/- +/- 4.79; chromium 1.64 +/- 1.47; cadmium 1.61 +/- 1.62; mercury 0.36 +/- 0.39 . Chlorinated carbohydrates and triazine and diazine herbicides were present in the following amounts (mg per kg): HCB 0.0023 +/- 0.0021; Lindane 0.0058 +/- 0.0079; DDE 0.0048 +/- +/- 0.0024; DDT 0.0065 +/- 0.0015; Simazine 0.060 +/- 0.020; Atrazine 0.083 +/- 0.059; Prometryn 0.093 +/- 0.040; Chloridazon 0.036 +/- 0.008; Terbutryn 0.085 +/- 0.029 . The content of aflatoxin B1 was 12.89 +/- 9.31 micrograms per kg and the maximum amount of polychlorinated biphenyls was 8 mg per kg . Nitrovin was found out only in the dust of two halls: 4.0 and 7.9 mg per kg . The dust deposition also contained 21 genera and species of moulds, six species of mites, numerous saprophytic bacteria and, in some cases, Staphylococcus aureus . For the time being, no viruses have been detected in the dust samples.

Am J Surg, 1986 Apr, 151(4), 437 - 42
Oral neomycin and erythromycin compared with single-dose systemic metronidazole and ceftriaxone prophylaxis in elective colorectal surgery; Weaver M et al.; A prospective randomized trial was performed to compare oral neomycin and erythromycin with single-dose intravenous metronidazole and ceftriaxone in elective colorectal surgery . The study was discontinued after 60 patients were entered . The overall rate of infection was 41 percent in the oral neomycin and erythromycin group (n = 29) compared with 9.6 percent in those who received intravenous metronidazole and ceftriaxone (n = 31) (p less than 0.01) . Infections in the oral group were principally due to resistant Staphylococcus aureus, Bacteroides fragilis, and Escherichia coli . Preoperative administration of oral neomycin and erythromycin was associated with a significant reduction of Escherichia coli counts (1 X 10(7) to 3 X 10(5) organisms/ml, p less than 0.05) compared with the intravenous group, but there was no significant reduction in the counts of Bacteroides fragilis (2 X 10(8) to 1 X 10(7) organisms/ml) and there was an increase in the counts of Clostridia (2 X 10(4) to 1 X 10(6) organisms/ml) . These results indicate that single-dose systemic prophylaxis with appropriate antibiotics is superior to oral neomycin and erythromycin.

J Clin Invest, 1986 Apr, 77(4), 1173 - 9
Recombinant interleukin 2 and gamma-interferon act synergistically on distinct steps of in vitro terminal human B cell maturation; Le thi Bich-Thuy et al.; The effects of recombinant interleukin 2 (IL-2) on the in vitro differentiation of human tonsillar B cells which were not preincubated with Staphylococcus aureus Cowan I or with anti-human IgM were investigated . IL-2 was shown to induce the generation of Ig-containing cells in a dose-dependent fashion from 2.5 to 2,500 U IL-2/ml . Conversely, the quantities of Ig secreted in the culture supernatant were found in the majority of experiments to peak at 25 U/ml . The possible presence, in cultures stimulated with IL-2, of cells that were capable of synthesizing Ig but that did not secrete the Ig they have produced was investigated . Among a number of factors tested, we found that gamma-interferon, which did not trigger in vitro B cell differentiation when used alone, can induce an increased secretion of Ig without noticeable change in the number of Ig-containing cells in cultures stimulated with IL-2 . The possibility that gamma-interferon and IL-2 act on subsequent steps of in vitro B cell differentiation is discussed.

Contraception, 1986 Apr, 33(4), 395 - 9
Effect of vaginal contraceptive sponges on growth of toxic shock syndrome-associated Staphylococcus aureus in vitro; Stumpf PG et al.; Toxic Shock Syndrome (TSS) is associated with certain toxin-producing strains of Staphylococcus aureus (TSS-S aureus), and with the use of some vaginal devices such as tampons or contraceptive diaphragms . The present study was designed to examine the effect of Nonoxynol-9 (N-9), and of a newly-approved vaginal contraceptive sponge (VCS) containing N-9 on the growth of TSS-S aureus in vitro . Flasks containing culture media inoculated with TSS-S aureus were incubated at 37 degrees C for 30 hours, in the presence or absence of either a VCS, or N-9 alone . At 0.5, 1, 2, 6, and 12 hours, there was suppression of TSS-S aureus colony counts in media containing VCS, compared to control . Colony counts from media containing N-9 demonstrated suppression at 0.5, 1, 2, and 6 hours . After incubation as long as 30 hours, colony counts from VCS-containing media approached, but did not exceed, counts from control media . From these data, it is concluded that this VCS containing N-9 does not enhance the growth of TSS-S aureus in vitro . Instead, an inhibition of bacterial growth for at least 12 hours is observed in media containing VCS, consistent with a bacteriostatic effect . If such an effect is also present in vivo, it would suggest that this type of VCS is unlikely to increase the risk of TSS.

Clin Exp Immunol, 1986 Apr, 64(1), 181 - 7
Mitogenic response of neoplastic B cells: comparison of reactivity to Staphylococcus aureus Cowan I and anti-immunoglobulin antibodies; Tamaki T et al.; Neoplastic B cells from two patients with hyperleucocytic hairy cell leukaemia (HCL) and 19 patients with chronic lymphocytic leukaemia of B cell Type (B-CLL) were investigated to examine the mitogenic responses to the F(ab')2 fraction of anti-human immunoglobulins (anti-Igs) and Staphylococcus aureus Cowan I (STA) . Neoplastic cells from both HCL patients lacked surface Tac antigen . Mononuclear cells from the two HCL patients strongly responded to both anti-Igs and STA as measured by 3H-thymidine incorporation in vitro . Although the mononuclear cells from two patients with B-CLL showed high response to STA, cells from none of the patients with B-CLL responded to anti-Igs . Mononuclear cells as well as T-cell-depleted fractions from the two HCL patients showed a strong proliferative response by anti-gamma chain antibody (anti-gamma) and the mononuclear cells from one of the patients were also induced to proliferate by anti-delta, whereas those from normal subjects responded only to a high concentration of anti-mu . Based on the difference in reactivity to anti-Ig, it is suggested that the HCL cells in this study originate from a subset equivalent to 'memory' B cells, whereas the B-CLL cells originate from a subset equivalent to 'virgin' B cells.

J Appl Bacteriol, 1986 Apr, 60(4), 277 - 87
Plasmids of Staphylococcus aureus associated with live and processed poultry; Thompson JK et al.; A series of 23 strains of Staphylococcus aureus originally isolated from processed poultry was screened for the presence of plasmids . Plasmids were more common in strains of Staph . aureus characteristically associated with live poultry than with strains endemic in poultry plants and strains of human origin . Two plasmids with sizes of 1.65 and 18.2 kilobase pairs (kBp) were present in three strains considered typical of Staph . aureus forma specialis 'altilis' and two plasmids with sizes of 1.65 and 17 kBp were present in three of four strains of Staph . aureus var . gallinae . A 1.65 kBp plasmid was present in all seven strains of these poultry biotypes and in three of 14 'endemic' strains . All the 1.65 kBp plasmids were shown by blot hybridization to share sequence homology . There was also some sequence homology between the 18.2 kBp and 17 kBp plasmids . These results were supported by restriction enzyme digest analyses . A study of cured derivatives of strain PS221 f.sp . 'altilis' suggested that the 18.2 kBp plasmid encoded the genetic determinant(s) responsible for caseolysis . Both the 1.65 and the 18.2 kBp plasmids also exerted an effect on the production of acid from lactose . In no other characteristic did cured strains resemble the plasmid-free 'endemic' strains . This was therefore consistent with the notion that the genetic determinants associated with the cultural characteristics of endemic strains are chromosomally located.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Apr, (4), 71 - 5
{Action of plant extracts on the natural immunity indices of animals}; Zakharova NS et al.; The influence of extracts from oak bark, St . John's-wort leaves and pine buds on natural immunity characteristics of mice has been studied . The injection of these extracts into mice has been found to enhance their resistance to infection with Staphylococcus aureus and Bordetella pertussis virulent cultures, to decrease the enzymatic activity of 5'-nucleotidase in the peritoneal exudate macrophages of mice and to increase the level of lysozyme in their blood . The action of these extracts has proved to depend on their dosage and the time of observation.

Diagn Microbiol Infect Dis, 1986 Apr, 4(4), 327 - 33
Rapid identification of coxsackie B viruses after immunoprecipitation and nucleic acid hybridization; Tracy S et al.; To circumvent the difficulties in concentrating sufficient virus from a clinical or environmental sample for detection and identification, we have used immunoprecipitation to rapidly concentrate coxsackie B viruses from both large and small sample volumes . Antiviral serum and killed Staphylococcus aureus cells as a protein A source were used to bind and collect the virus . Radioactively-labeled viral nucleotide sequences were used to identify the collected virus by nucleic acid hybridization . The technique is applicable to the rapid concentration of dilute viruses from extremely large sample volumes as well as the samples from which virus isolation can be difficult . the process requires less than 2 days to complete and should be adaptable to virus identification by cell culture or other standard means as well.

J Infect Dis, 1986 Apr, 153(4), 670 - 5
Transformation of a plasmid encoding an adhesin of Staphylococcus aureus into a nonadherent staphylococcal strain; Dunkle LM et al.; Plasmid DNA was isolated and purified from a strain of Staphylococcus aureus demonstrating adherence to human epithelial cells, as determined by an assay quantitating adherence of radiolabeled S . aureus to HeLa cells in tissue culture . Other phenotypic characteristics of the donor strain are resistance to ampicillin and absence of hemolysis . A 23.5-kb (kilobase) plasmid was transformed into a nonadherent, ampicillin-sensitive, beta-hemolytic recipient of S . aureus, rendering it both ampicillin-resistant and adherent to a degree approaching that of the donor strain . Plasmid analysis of the donor and transformant strains revealed three identical EcoRI fragments of 7.6 kb, 6.5 kb, and 2.2 kb, together with a 3.6-kb EcoRI fragment in the transformant that demonstrated homology with the last 7.2-kb fragment in the donor . We conclude that an adhesin of S . aureus is encoded by this 23.5-kb penicillinase-encoding plasmid and that techniques of molecular genetics may be utilized to clarify the mechanisms of adherence of S . aureus.

J Invest Dermatol, 1986 Apr, 86(4), 390 - 3
Qualitative and quantitative changes in cutaneous bacteria associated with systemic isotretinoin therapy for acne conglobata; Leyden JJ et al.; Quantitative cultures in 40 patients treated with systemic isotretinoin demonstrated a significant reduction in the anaerobic diphtheroid, Propionibacterium acnes within one month of therapy and a continued suppression during 5 months of treatment . This reduction persisted after discontinuation of isotretinoin therapy despite a return of sebum excretion to pretreatment levels . Surface aerobic bacteria showed a significant reduction in the total number of organisms and significant qualitative changes . Gram negative bacteria were sharply reduced in both the anterior nares and skin while Staphylococcus aureus recovery increased significantly.

Poult Sci, 1986 Apr, 65(4), 696 - 703
Binding of chicken, bovine, and rabbit immunoglobulins by avian, bovine, and human strains of Staphylococcus aureus; Schwan WR et al.; Sixty-six strains of Staphylococcus aureus of avian, bovine, and human origin were tested for their ability to bind chicken, bovine, and rabbit immunoglobulin G (IgG) . A microtitration plate hemagglutination assay and a direct-tube enzyme immunoassay were used to determine qualitative differences . Twice as many chicken and bovine S . aureus isolates than human strains reacted positively to chicken IgG . Mean binding values of chicken IgG were also twice as high for chicken and bovine S . aureus isolates when compared with human-derived strains . Isolates of bovine and human origin displayed a high affinity for bovine IgG and rabbit IgG, whereas few isolates from chickens bound substantial quantities of the nonavian IgG species . These results demonstrate that preparations of staphylococcal protein A with affinities for immunoglobulins from poultry and other animals can be obtained by screening large numbers of isolates, especially those obtained from an animal species, such as chickens, that is the same as the source of the immunoglobulin one wishes to bind.

J Antimicrob Chemother, 1986 Apr, 17 Suppl B, 49 - 52
Comparative efficacy of pefloxacin and six other antimicrobial agents on Staphylococcus aureus experimental abscesses; Rolin O et al.; Pefloxacin (Pef) is a new quinolone which has been shown to have good in-vitro activity against Staphylococcus aureus, and to reach high tissue concentrations . Its efficacy was compared to that of 2 quinolone derivatives, norfloxacin (Nor) and ciprofloxacin (Cip) and to that of methicillin (Meth), cephalothin (Cep), pristinamycin (Pri) and vancomycin (Van), in an experimental model of S . aureus abscesses . Mice challenged with an intramuscular thigh injection of a calibrated inoculum developed local abscesses . Three S . aureus strains (with different antibiotic resistance profiles (Pase-, Pase+, MethR)) were used . In this model, the antibiotics showing the best ED50 following oral administration, against all three strains were Pef greater than Cip greater than Pri greater than Nor, by subcutaneous administration for the Pase- strain Pef = Cip greater than Cep greater than Van; for the Pase+ strain Pef = Cip greater than Van greater than Meth and for the Pase+ MethR strain: Pef = Cip greater than Van greater than Cep . These data indicate that pefloxacin and ciprofloxacin are highly active in vivo against various strains of S . aureus, and appear to be more potent than norfloxacin, vancomycin, cephalothin and methicillin.

Oral Surg Oral Med Oral Pathol, 1986 Apr, 61(4), 413 - 7
Dental treatment and late prosthetic joint infections; Jacobson JJ et al.; Hospital and dental charts of 2,693 patients in whom total prosthetic joints had been placed at the Veterans Administration Hospitals of Ann Arbor and Allen Park, Michigan, as well as at The University of Michigan Hospital, were analyzed . Of the thirty (1.1%) late prosthetic joint infections (greater than 6 months after placement), only one (0.04%) could be temporally associated with dental treatment . A Fisher's exact test of the data reflected that dental treatment in this population did not increase the incidence of late prosthetic joint infections (p value is 0.0005) . Nine of the thirty late infections occurred in insulin-dependent diabetic patients and patients on long-term immunosuppressive therapy . An analysis of the organisms isolated from the late infections shows that 54% where Staphylococcus epidermidis and Staphylococcus aureus . These data do not support the practice of prescribing prophylactic antibiotic coverage of prosthetic hip and knee joints prior to all dental therapy . Rather, use of antibiotics during dental treatment appears warranted only if a chronic bacteremia is anticipated or where a predisposing systemic condition may exist.

J Neurochem, 1986 Apr, 46(4), 1263 - 71
Cyclic nucleotide phosphodiesterase activity in bovine brain coated vesicles; Silva WI et al.; Cyclic AMP phosphodiesterase activity in bovine brain coated vesicles displayed a Km of approximately 22 microM for cyclic AMP, a Vmax of 3.2 nmol/min/mg protein, and a Hill coefficient of 1.5, suggesting positive cooperativity . The enzyme activity was stimulated by cyclic GMP with maximal indexes of stimulation ranging between 40 and 300% . Both basal and stimulated phosphodiesterase activities were immunotitrated with polyclonal antibodies against clathrin attached to heat-inactivated, formaldehyde-fixed Staphylococcus aureus cells . The main form of phosphodiesterase activity present in the immunoprecipitated brain coated vesicle preparation also is stimulated by cyclic GMP . The allosteric behavior was modulated by cyclic GMP . All of these properties are typical of type II or cyclic GMP-sensitive phosphodiesterases in addition to their calcium and calmodulin independence . Competition experiments with a series of phosphodiesterase inhibitors, papaverine, 1-methyl-3-isobutylxanthine, and theophylline, showed inhibition of cyclic AMP hydrolysis . Trifluoperazine was inactive at the highest concentration used, 100 microM . These compounds also inhibited the cyclic GMP-stimulated cyclic AMP hydrolysis with trifluoperazine practically inactive . At 5 microM cyclic AMP none of the inhibitors was seen to stimulate the cyclic AMP hydrolytic activity . The presence of an enzyme for the breakdown of cyclic nucleotides in brain coated vesicles may suggest a role for these second messengers in the in vivo functions of this organelle.

Biochimie, 1986 Apr, 68(4), 531 - 41
Isolation and characterization of three monoclonal antibodies to human serum low density lipoprotein apoprotein B; Aggerbeck LP et al.; Human serum low density lipoprotein (LDL) is a large (Mr = 2-3 X 10(6), complex particle composed of lipid, protein and carbohydrate . We obtained about 40 mouse spleen-myeloma hybrid cell lines which produce antibodies against LDL . Three of them, SC2, SC3 and SC10, have been cloned and subcloned and their antibody products characterized . They recognize three non-overlapping epitopes in native LDL . Two of them, SC3 and SC10, also are capable of recognizing very low density lipoprotein, (VLDL), whereas SC2 reacts only weakly with VLDL . All three antigenic determinants remain intact, and accessible to antibodies on the LDL protein apo B, prepared by delipidation in a 'non-denaturing' detergent, sodium deoxycholate . However, apo B prepared by organic solvent, ether-ethanol, or sodium dodecyl sulfate (SDS) delipidation, while reacting strongly with SC10, is only poorly recognized by SC2 or SC3 . Proteolysis of LDL with trypsin, chymotrypsin, Staphylococcus aureus protease, papain or thermolysin gives, in each case, several non-identical protein fragments which are separable by SDS-polyacrylamide gel electrophoresis . Upon immunoblotting, some of these fragments are now recognized by either SC3 or SC10 but not SC2, some are recognized by both SC3 and SC10, and others are immunologically unreactive . The protein bands that are separated by SDS gel electrophoresis are composed of several non-identical fragments and contain the antigenic sites to differing degrees . Some of the immunologically reactive fragments do not appear to contain carbohydrate . Reduction and carboxymethylation do not destroy the immunoreactivity of LDL toward any of the antibodies; however, modification of lysine residues by citraconic anhydride markedly diminishes the reactivity of LDL toward SC3 . It is likely that the two antibodies SC3 and SC10 are directed against different linear amino acid sequences or very stable domains, whereas the third, SC2, is directed against a more fragile conformational domain of apo B.

Thorac Cardiovasc Surg, 1986 Apr, 34(2), 94 - 7
Intrathoracic surgery for retained endocardial electrodes; Jarvinen A et al.; Intrathoracic surgery for retained endocardial electrodes was performed in 12 patients; the indication for electrode removal was Staphylococcus aureus or epidermidis infection in 11 patients and malfunction in one patient . Two operations had to be performed on emergency basis . One was carried out because of myocardial rupture and cardiac tamponade after a tightly fixed electrode lead had been pulled out . The other patient was operated on because of ventricular arrhythmia arising from a malfunctioning lead which had slipped back completely after transsection into the right ventricle . Ten patients underwent elective surgery; cardiopulmonary bypass was needed in 6 of these . In one case a mitral prosthesis was replaced because of infection . Radical treatment is recommended for pacemaker infections and the removal of electrodes should be considered even if intrathoracic surgery is required.

Eur J Biochem, 1986 Apr 1, 156(1), 9 - 14
Molecular cloning of cDNA for rat mitochondrial 3-hydroxyacyl-CoA dehydrogenase; Amaya Y et al.; Messenger RNA for 3-hydroxyacyl-CoA dehydrogenase, a mitochondrial matrix enzyme of fatty acid beta-oxidation, was purified from livers of di(2-ethylhexyl)phthalate-treated rats by immunoadsorption of hepatic free polysomes to fixed cells of Staphylococcus aureus and enrichment for poly(A)-rich RNA by oligo(dT)-cellulose chromatography . Plasmid cDNA was constructed from this poly(A)-rich RNA by a modification of the method of Okayama and Berg and was transformed into the Escherichia coli DH1 strain . Plasmids containing cDNA sequences coding for 3-hydroxyacyl-CoA dehydrogenase were screened by differential colony hybridization, and were identified by hybrid-arrested translation and hybrid-selected translation . Plasmid pHADH-1, which contains a 1400-base-pair insert, hybridized to rat 3-hydroxyacyl-CoA dehydrogenase mRNA with a length of 1700 bases . Determination of the dehydrogenase mRNA by in vitro translation and dot-blot analysis with the cDNA probe showed that the induction of the enzyme in rat liver by di(2-ethylhexyl)phthalate could be attributed to an increase in the mRNA concentration.

Biochim Biophys Acta, 1986 Mar 27, 856(1), 91 - 100
Lytic effects of melittin and delta-haemolysin from Staphylococcus aureus on vesicles of dipalmitoylphosphatidylcholine; Yianni YP et al.; The effects of the lytic peptides, melittin and delta-haemolysin, are compared in vesicles of gel-phase dipalmitoylphosphatidylcholine (DPPC), using calcein as trapped marker . At low concentration, both toxins cause vesicles to lose contents in 5 mM phosphate buffer near neutral pH, with melittin being the more active . As phosphate concentration is increased, the kinetics of melittin-induced leakage change from a slow, sustained loss to a rapid 'burst' of leakage when melittin is present mainly as tetramer in solution, under conditions where it is reported to lose haemolytic activity towards erythrocytes . At low phosphate concentration, the leakage induced by delta-haemolysin is preceded by a lag phase, though fluorescence measurements show that binding of toxin is rapid . At higher phosphate concentration, the toxin binds rapidly to vesicles, but causes no leakage of entrapped calcein . Steady-state fluorescence spectra show no obvious differences in tryptophan emission for delta-haemolysin bound to lipid in high- or low-phosphate buffer . Spin-label fluorescence-quenching studies show that the single tryptophan residue of delta-haemolysin is buried within the lipid bilayer at all phosphate concentrations used . In gel-phase DPPC, delta-haemolysin shows no tendency to cause vesicle aggregation over several hours, as judged by light scattering, though a slow non-linear effect is seen above the lipid phase transition temperature . These effects are contrasted with those of melittin under similar conditions.

Schweiz Med Wochenschr, 1986 Mar 15, 116(11), 331 - 5
{Significance of the determination of teichoic acid antibodies in the diagnosis of Staphylococcus aureus bacteremia}; Herzog C et al.; Sera of 47 patients with serious Staphylococcus aureus infections, 55 patients with septicemias caused by other organisms and 147 healthy controls were examined for teichoic acid antibodies by ELISA, counterimmunoelectrophoresis (CIE) and immunodiffusion (ID) . The specificity and the predictive value of positive results for metastatic-septic complications were 100% each for ELISA, 93%/88% for CIE and 87%/80% for ID . The low sensitivity with all three test methods of 50-60% relates to single sera . The indication for prolonged antibiotic treatment should therefore be derived only from the analysis of consecutive serum samples . With 12% of the controls above the cut-off, the ID test seems not a very good screening test.

Biochem Pharmacol, 1986 Mar 15, 35(6), 1001 - 4
In vitro {3H}-erythromycin binding to Staphylococcus aureus; Barre J et al.; Characteristics of erythromycin binding to Staphylococcus aureus were determined by using kinetics and equilibrium binding experiments . Both methods yielded identical values of the dissociation constant, i.e . 0.1 muM . This value was in accord with that found with a bacterial extract of ribosomes which are the organelles where erythromycin exerts its action . This good agreement shows that the dissociation constant of erythromycin determined with intact bacteria is a good reflect of specific bacterial receptors of macrolides, i.e . ribosomes . In addition, mechanism of uptake of the antibiotic by Staphylococcus aureus was investigated . Passive diffusion process was shown to be mainly responsible for this phenomenon.

Biochem J, 1986 Mar 15, 234(3), 665 - 70
Trypsin-catalysed formation of pig des-(23-63)-proinsulin from desoctapeptide-(B23-30)-insulin; Kubiak T et al.; Incubation of pig desoctapeptide-(B23-30)-insulin with trypsin in solvent systems consisting of dimethyl sulphoxide, butane-1,4-diol and Tris buffer resulted in the formation of an extra peptide bond between Arg-B22 and Gly-A1 in the DOPI molecule . This DOPI derivative can also be regarded as pig des-(23-63)-proinsulin . The structure of the new, previously unreported, proinsulin analogue was determined on the basis of amino acid analysis, dansylation and digestion with Staphylococcus aureus V8 proteinase . Receptor-binding ability of des-(23-63)-proinsulin was 20% of that of pig desoctapeptide-(B23-30)-insulin and 0.02% of that of pig insulin.

Vet Rec, 1986 Mar 15, 118(11), 290 - 1
Antimicrobial drug susceptibility of Staphylococcus aureus isolated from bovine mastitis; Craven N et al.; Isolates of Staphylococcus aureus (106) from bovine mastitis were tested for susceptibility to antimicrobial agents . beta-lactamase was produced by 69.8 per cent of isolates and 7.5 per cent were resistant to streptomycin (minimum inhibitory concentration more than 32 micrograms/ml) . Resistance to other agents was rare . Intrinsic resistance or tolerance to beta-lactam antibiotics was not found.

J Biol Chem, 1986 Mar 15, 261(8), 3894 - 900
Purification and characterization of the human platelet alpha 2-adrenergic receptor; Regan JW et al.; Human platelet alpha 2-adrenergic receptors have been purified approximately 80,000-fold to apparent homogeneity by a five-step chromatographic procedure . The overall yield starting from the membranes is approximately 2% . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of radioiodinated protein from purified receptor preparations shows a single major band of Mr 64,000 . The specific binding activity of the alpha 2-adrenergic receptor after four chromatographic steps is 14.5 nmol/mg protein . This value is consistent with the expected theoretical specific activity (15.6 nmol/mg) for a protein with a molecular mass of 64,000 daltons if it is assumed that there is one ligand-binding site/receptor molecule . The purified protein can be covalently labeled with the alkylating alpha-adrenergic ligand, {3H}phenoxybenzamine . This labeling is specific, and it shows that the Mr 64,000 protein contains the ligand binding site of the alpha 2-adrenergic receptor . In addition, the competitive binding of ligands to the purified receptor protein shows the proper alpha 2-adrenergic specificity . The alpha 2-adrenergic receptor contains an essential sulfhydryl residue . Thus, exposure of the purified receptor to the sulfhydryl-specific reagent, phenylmercuric chloride, resulted in an 80% loss of binding activity . This loss of binding activity was prevented when exposure to phenylmercuric chloride was done in the presence of alpha 2-adrenergic ligands, and it was reversed by subsequent exposure to dithiothreitol . Partial proteolysis of purified alpha 2-adrenergic receptors was obtained with Staphylococcus aureus V-8 protease, alpha-chymotrypsin, and papain . In a comparison with purified beta 2-adrenergic receptors, no common partial proteolytic products were found.

Biochemistry, 1986 Mar 11, 25(5), 1078 - 82
Development of a radioimmunoassay for quantitation of calregulin in bovine tissues; Khanna NC et al.; Experimental conditions are described for a convenient and simple one-step method for radioimmunoassay (RIA) of the calcium binding protein calregulin {Waisman, D.M., Salimath, B.P., & Anderson, M.J . (1985) J . Biol . Chem . 260, 1652-1660} . The radioimmunoassay utilizes 125I-labeled calregulin and pansorbin cells (Staphylococcus aureus) coated with rabbit anti-calregulin antibody . Binding equilibrium is attained in 30 min, and the total time of the assay is 1 h . The assay is sensitive to about 30 fmol of calregulin . Calregulin was quantitated in various bovine tissue extracts and was detected in all tissues except erythrocytes . It was present in particularly high amounts in pancreas (540 micrograms/g of tissue), liver (375 micrograms/g of tissue), and testis (256 micrograms/g of tissue) . While about 80% of the total tissue calregulin is soluble, about 20% of this protein was found to be associated with particulate fractions . Unmasking of particulate calregulin required the presence of nonionic detergent . Gel permeation chromatography of bovine brain 100 000 g supernatant in the presence or absence of calcium has resolved a single peak of calregulin by RIA . In addition, the distribution of calregulin in the soluble or particulate fraction of bovine brain was unaffected by the presence or absence of calcium during homogenization, suggesting that calregulin does not form either calcium-dependent or calcium-independent association with soluble or particulate proteins . These results identify calregulin as a major tissue Ca2+ binding protein.

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