Staphylococcus aureus

Nord Vet Med, 1983 Dec, 35(12), 460 - 4
Antimicrobial drug susceptibility of Staphylococcus aureus strains isolated from bovine milk; Franklin A et al.; The minimum inhibitory concentration (MIC) of ten antimicrobial drugs for 287 S . aureus strains recently isolated from bovine mastitic milk in different herds all over Sweden was determined . The minimum bactericidal concentration (MBC) of benzylpenicillin for 20 strains was determined . Thirty strains (10%) produced beta-lactamase . All strains were susceptible to oxacillin and neomycin, and more than 90% to streptomycin, trimethoprim-sulphamethoxazole chloramphenicol, erythromycin and tetracycline, whereas all were resistant to sulphamethoxazole . None of 20 strains investigated was tolerant to benzylpenicillin . However, S . aureus strains, isolated from bovine milk, should be tested for beta-lactamase production prior to treating mastitic cases with beta-lactam drugs.

Biochem J, 1983 Dec 1, 215(3), 525 - 9
Identification by n.m.r . spectroscopy of a stable intermediate structure in the unfolding of staphylococcal beta-lactamase; Thomas RM et al.; The unfolding of beta-lactamase (penicillinase) from Staphylococcus aureus by guanidinium chloride was followed by using n.m.r . spectroscopy . On the basis of the observation of resonances corresponding to histidine, tyrosine and other amino acid side chains, the existence of a stable partially folded species was demonstrated . These experiments provide detailed characterization of the intermediate that confirms and extends previous characterization by absorption and c.d . spectroscopy and by flow properties . In addition, they show that residues in the N-terminal third of the molecule are affected by the native-to-intermediate transition . Persistent non-equivalence of the two imidazole C2 proton resonances at high guanidinium chloride concentrations is discussed in terms of local sequence effects on the chemical shift.

Infect Immun, 1983 Dec, 42(3), 1013 - 6
Soluble peptidoglycan from Staphylococcus aureus is a murine B-lymphocyte mitogen; Babu UM et al.; Soluble peptidoglycan from Staphylococcus aureus has been shown to be capable of causing murine B lymphocytes from the spleen to proliferate and to secrete immunoglobulins in both an in vitro and an in vivo assay . The optimal concentration in vitro was between 33 and 100 micrograms/ml . A 3-day incubation with soluble peptidoglycan was more stimulatory than was a 1- or 2-day incubation . Removal of most of the T lymphocytes with anti-theta serum did not result in any significant change in the mitogenic activity of soluble peptidoglycan on the remaining B cells.

J Antimicrob Chemother, 1983 Dec, 12 Suppl D, 79 - 87
Comparative imipenem treatment of Staphylococcus aureus endocarditis in the rat; Baumgartner JD et al.; The efficacy of imipenem alone or in association with gentamicin against Staphylococcus aureus experimental endocarditis was compared to the efficacy of cloxacillin alone or in association with gentamicin . Parenteral treatment was started 24 h after intravenous bacterial challenge of rats with catheter-induced aortic valve vegetations . The cloxacillin MIC and MBC for Staph . aureus were 0.125 and 32 mg/l and the imipenem MIC and MBC 0.008 and 8 mg/l, respectively . In-vitro killing curves showed a synergistic effect between cloxacillin and gentamicin, and an additive effect between imipenem and gentamicin . Only large doses of cloxacillin (400 mg/kg tid) (producing serum levels above those obtained after intravenous injection of 2 g in man) achieved results comparable to those of imipenem 80 mg/kg tid (producing serum levels similar to those obtained after an intravenous dose of 750 mg in man) in reducing the bacterial numbers in vegetations after 3 and 5 days of treatment . There was a significantly greater reduction of bacterial numbers in vegetations after treatment with the association of cloxacillin and gentamicin than with cloxacillin alone . In contrast, the addition of gentamicin to imipenem did not improve significantly the results of treatment with imipenem alone, but imipenem alone was as good as the combination cloxacillin and gentamicin after 5 days of treatment . We conclude that imipenem is a highly bactericidal drug in this animal model, worth considering for clinical trials in the treatment of Staph . aureus infections.

Zh Mikrobiol Epidemiol Immunobiol, 1983 Dec, (12), 30 - 3
{Mechanisms of the bactericidal action of hydrogen peroxide}; Samoilenko II et al.; The comparative study of some aspects of the bactericidal action of H2O2 on Escherichia coli, Staphylococcus aureus and Bacterium subtilis wild-type cells and their mutants with lesions in the systems of the reparation of DNA has been carried out . Hydrogen peroxide has been shown to cause disturbances in the structure and permeability of the cell wall, the cytoplasmic membrane, as well as to induce ribosomal lesions and the ruptures of bacterial DNA . The activity of the systems responsible for the reparation of lesions in the cell genome plays an important role in the resistance of bacteria to H2O2.

J Gen Microbiol, 1983 Dec, 129 ( Pt 12), 3603 - 10
Immunochemical studies of Staphylococcus aureus Oeding-Haukenes antigen a5: a phosphorus-containing polysaccharide; Ndulue AN et al.; Antigen a5 was isolated from strain 830 of Staphylococcus aureus by autolysis in phosphate buffer followed by alcohol precipitation . Purification was principally achieved by affinity chromatography on wheat germ agglutinin ultrogel and on anti-S . aureus teichoic acid immunosorbent . The a5 antigen was weakly immunogenic in rabbits . Chemical analysis showed that a5 is a teichoic acid composed of ribitol phosphate, N-acetylglucosamine and alanine . It has similar physico-chemical properties to the wall beta-N-acetylglucosamine ribitol teichoic acid of S . aureus but is serologically distinct.

Zh Mikrobiol Epidemiol Immunobiol, 1983 Dec, (12), 60 - 4
{Neutrophil phagocytic activity of the peripheral blood in experimental keratoconjunctivitis in a sensitized macroorganism}; Milovidova OV et al.; The study of the characteristics of the phagocytic activity of peripheral blood neutrophils (the activity and intensity of phagocytosis, the index of its completeness) in the sensitized organism in experimental keratoconjunctivitis caused by Staphylococcus aureus and Escherichia coli has revealed a decrease in the phagocytic function of neutrophils . Still more pronounced suppression of the ingestive and digestive activity of leukocytes has been observed in cases of the combined action of bacterial allergens and benzylpenicillin potassium, which probably accounts for the ineffectiveness of the penicillin treatment of bacterial keratoconjunctivitis.

Proc Natl Acad Sci U S A, 1983 Dec, 80(23), 7109 - 12
Protease-sensitive regions in myosin subfragment 1; Applegate D et al.; Proteolytic digestions of myosin subfragment 1 (S-1) with elastase, subtilisin, papain, thermolysin, and Staphylococcus aureus protease reveal that the two trypsin-sensitive regions in S-1 have broad protease susceptibility . The cleavage of S-1 by these enzymes yields products that correspond within 1-2 kilodaltons (kDa) to the 25-, 50-, and 20-kDa fragments produced by trypsin . Papain and thermolysin cut preferentially at the 26-kDa/70-kDa junction, whereas elastase, subtilisin, and S . aureus protease cleave both the 26-kDa/70-kDa and 75-kDa/22-kDa junctions in S-1 . Binding of actin to S-1 decreases the rate of all proteolytic reactions in the 95-kDa heavy chain . The protection of the 26-kDa/70-kDa junction by actin is greatest against papain and thermolysin attack . The reaction times of elastase, subtilisin, and S . aureus protease with S-1 increase 2-fold in the presence of actin . However, in contrast to similar reactions with trypsin, they proceed at both junctions and lead to formation of the 50- and 22-kDa fragments . The cleavage of the 22-kDa/50-kDa junction by elastase increases the Km value for the actin-activated ATPase . The presence of the two protease-sensitive regions in S-1 is consistent with a three-domain structure of the myosin head and may have important implications to the mode of intersite communication in this protein.

Acta Pathol Microbiol Immunol Scand {C}, 1983 Dec, 91(6), 355 - 9
Effect of temperature on polymorphonuclear leukocyte function; Johansen KS et al.; The effect of temperature on the function of polymorphonuclear leukocytes (PMN) has been investigated in vitro . Increases in temperature from 37 degrees C to 40 degrees C progressively increased chemiluminescence (CL) responses by PMN after stimulation by Staphylococcus aureus, zymosan or phorbol myristate acetate (PMA) while increases above 40 degrees C decreased these functions . Temperature increases from 37 degrees C to 40 degrees C also produced increased PMN bactericidal activity against S . aureus . In contrast, similar increases in temperature did not change superoxide production by PMN stimulated by PMA . Incubation of PMN at the various temperatures did not cause release of LDH indicating that damage to PMN was not the cause of reduced PMN chemiluminescence and bactericidal activity seen within the temperature range studied . The discrepancy between the influence of temperature on PMN chemiluminescence and bactericidal activity of PMN compared to superoxide anion production by PMN suggests that superoxide anion production may not be solely, or at least directly, responsible for killing of bacteria . Careful temperature control is needed when assaying PMN function . Febrile responses up to 40 degrees C may play a beneficial role in host defense.

Acta Pathol Microbiol Immunol Scand {B}, 1983 Dec, 91(6), 425 - 9
Population analysis in strains of Staphylococcus aureus and Staphylococcus epidermidis . II . Cefuroxime and cefotaxime; Hansen BG; Population analyses of susceptibility to cefuroxime and cefotaxime in penicillin-susceptible, penicillin-resistant, and methicillin-resistant strains of Staphylococcus aureus (S . aureus) and Staphylococcus epidermidis (S . epidermidis) were carried out . All strains were clinical isolates . Both antibiotics were shown to be more penicillinase-stable than cephalothin in studies of the penicillin-resistant strains of S . aureus, but less stable than methicillin . The studies of penicillin-resistant strains of S . epidermidis showed no differences in penicillinase-stability between cephalothin and the new cephalosporins . From the methicillin-resistant strains of S . aureus and S . epidermidis it was possible to select highly-resistant mutants against both antibiotics with a frequency of c . 10(-5) although MIC determinations had shown the strains to be susceptible.

Proc Natl Acad Sci U S A, 1983 Dec, 80(24), 7538 - 41
Vitamin D metabolites change the phenotype of monoblastic U937 cells; Dodd RC et al.; U937 is a human-derived lymphoma cell line that has monoblastic properties and high-affinity receptors for 1 alpha,-dihydroxyvitamin D3 . Incubation of these cells with the vitamin D metabolite at 10 nM for 5 days produced marked stimulation in adherence and ingestion of Staphylococcus aureus (645% of control) and of C3b receptor (CR1) expression (292% of control) and a slight increase in hexose monophosphate shunt activity without changing cell growth rates or Fc fragment receptor expression . The changes in cellular association of S . aureus and the CR1 were detected as early as 48 hr of incubation and peaked between 3 and 5 days . Similar changes in the CR1 were induced by 25-hydroxy- and 24,25-dihydroxyvitamin D3 at micromolar concentrations . Dexamethasone, hydrocortisone, and progesterone had no effect on CR1 expression . U937 cells incubated in the presence of vitamin D metabolites exhibited a change in their phenotype . These results suggest that vitamin D metabolites may contribute to monocyte/macrophage differentiation.

Sem Hop, 1983 Dec 1, 59(44), 3087 - 8
{Selection of mutant rifampin-resistant Staphylococcus aureus during the therapeutic use of this antibiotic in combinations . 3 cases}; Dellamonica P et al.; Selection of rifampicin-resistant Staphylococcus aureus has been described in vitro and in vivo when this compound is given as monotherapy or orally . That this occurrence may be prevented by combination-antibiotic therapy is generally accepted . We report three cases of serious staphylococcal infection treated by a synergic association of rifampicin with either an aminoglycoside or vancomycin . Therapy failed as a result of selection of the same rifampicin-resistant Staphylococcus aureus (serotype and lysotype) . These observations may be explained by insufficient diffusion or inactivation of the other antibiotic in the infection site.

Am J Vet Res, 1983 Dec, 44(12), 2366 - 72
Suppression of neutrophil and lymphocyte function induced by a vaccinal strain of bovine viral diarrhea virus with and without the administration of ACTH; Roth JA et al.; Effects of a modified live vaccine (MLV) strain of bovine viral diarrhea virus (BVD) on lymphocyte and neutrophil function were determined in cattle with and without increased plasma cortisol (hydrocortisone) concentrations . Cattle were given MLV-BVD vaccine IM and intranasally . Cattle given ACTH received 200 IU every 12 hours for 10 doses . The MLV-BVD virus when administered alone caused no apparent clinical signs or body temperature response . Of 4 MLV-BVD-treated calves that were also given ACTH, 2 developed increased body temperature and respiratory distress . The MLV-BVD virus caused a decrease in circulating lymphocytes and neutrophils, whereas administration of ACTH and MLV-BVD induced a neutrophilia and lymphopenia . The MLV-BVD virus and ACTH when administered separately or in combination caused a depression of lymphocyte blastogenesis in response to selected mitogens . Neutrophils were separated from the peripheral blood and their function was evaluated, using the following procedures: (i) random migration under agarose, (ii) ingestion of 125I-labeled Staphylococcus aureus, (iii) quantitative nitroblue tetrazolium reduction, (iv) iodination, and (v) antibody-dependent cell-mediated cytotoxicity (ADCC) . The MLV-BVD virus produced a significant (P less than 0.05) suppression of neutrophil iodination and ADCC . Neutrophils from cattle given MLV-BVD virus and ACTH had enhanced random migration, enhanced S aureus ingestion, suppressed iodination, and suppressed ADCC activity.

Virology, 1983 Dec, 131(2), 315 - 27
The feline oncornavirus-associated cell membrane antigen (FOCMA) is related to, but distinguishable from, FeLV-C gp70; Snyder HW Jr et al.; The feline oncornavirus-associated cell membrane antigen (FOCMA) on the surface of feline lymphosarcoma (LSA) cells is defined as the target(s) recognized in immunofluorescence (IFA) tests by antibody in sera of cats relatively resistant to development of FeLV (feline leukemia virus) LSA and FeSV (feline sarcoma virus) fibrosarcoma . The specificities of antibodies in cat FOCMA-typing sera and the nature of the LSA antigens recognized were investigated in the present study . FOCMA sera obtained from viremic cats were separable into at least two classes : those which contained antibodies against the envelope glycoprotein (gp70) of subgroup C FeLV and those which did not contain antibodies against any subgroup of FeLV . The first class of sera could be further subdivided into three groups: those whose FOCMA reactivity could be completely absorbed, partially absorbed, or not absorbed by FeLV-C antigens . The second class of sera could be further subdivided into two groups: those whose FOCMA reactivity could be partially absorbed and those whose activity could not be absorbed by FeLV-C . The results indicate that the FOCMA reactivity exhibited by some viremic cat sera can be partially, if not entirely, attributed to antibodies not crossreactive with FeLV virion antigens . A consistent property of all FOCMA sera in this study is the ability to bind to 70-kDa proteins on the surface of LSA cells . Staphylococcus aureus V8 protease partial digest maps of 70-kDa proteins purified from 12 primary feline LSAs (five FeLV positive and seven FeLV negative) all showed 18-, 14-, and 10-kDa fragments . V8 maps of FeLV-C gp70 showed similarly sized fragments while the maps of the RD114, FeLV-A, and FeLV-B gp70s were distinct . However, in a subgroup-specific radioimmunoassay for FeLV-C gp70-related antigens, the LSA 70-kDa proteins were found to be serologically related to, but distinct from, FeLV-C gp70 . The results on the antigenic variations among LSA 70-kDa proteins and the antibodies which bind them are entirely consistent with previous studies indicating heterogeneity among FOCMA determinants.

Eur J Biochem, 1983 Dec 1, 137(1-2), 269 - 77
The primary structure of turkey muscle acylphosphatase; Camici G et al.; The complete primary structure of turkey muscle acylphosphatase has been determined . The sequence was derived from peptides obtained by digestion of the carboxymethylated protein with pepsin and thermolysin and by subdigestion of some of the cyanogen bromide fragments with trypsin and Staphylococcus aureus protease . Peptides were purified by preparative finger prints and/or preparative high-performance liquid chromatography . Sequencing of the various peptides was achieved by manual Edman degradation and by time-course analysis of amino acids released by carboxypeptidases . The amino-terminal blocking group (acetyl) was determined by fast atom bombardment mass spectrometry . This sequence was compared with that of horse muscle enzyme determined previously.

J Hosp Infect, 1983 Dec, 4(4), 331 - 7
Genetics of drug resistance in methicillin-resistant Staphylococcus aureus from Australian hospitals; Townsend DE et al.; The drug-resistance determinants in methicillin-resistant Staphylococcus aureus (MRSA) from three different hospitals in Eastern Australia have been examined . With one exception, all the isolates had chromosomal determinants for penicillinase and resistance to cadmium (Cd), mercury (Hg), phenyl mercuric acetate, methicillin, tetracycline, erythromycin, lincomycin and low level streptomycin . The strain which was the exception differed in that it did not have chromosomal resistance to Cd and lincomycin . In addition, the strains often contained plasmids which belonged to one of three categories: a small cryptic plasmid of either c . 1.4 Mdal, c . 1.7 Mdal or c . 1.9 Mdal; a chloramphenicol resistance plasmid of c . 2.8 Mdal; and a gentamicin resistance plasmid within the range of c . 15.3 to c . 28.5 Mdal . The predominant gentamicin-resistant plasmid in isolates from two hospitals had a molecular weight of c . 18 Mdal, whereas the isolates from the third hospital had a plasmid of molecular weight c . 15.3 Mdal . The only other gentamicin resistance plasmids detected were associated with penicillinase determinants . In one isolate, this corresponded to a plasmid of c . 19.6 Mdal and in the other to a plasmid of c . 28.5 Mdal . These results indicate that MRSAs which are prevalent in Eastern Australian hospitals are substantially different in the location of their drug resistance determinants to earlier strains reported in the literature.

Boll Ist Sieroter Milan, 1983 Nov 30, 62(5), 406 - 11
Staphylococcus aureus resistant to methicillin and gentamicin as a cause of outbreak of epidemic enteritis in a hospital; Scopetti F et al.; During the month of February 1982 in an orthopedic department (37 patients admitted), episodes of diarrhoea occurred in 9 patients, one of these resulting in the death of one patient . From the stool cultures of the patients methicillin/gentamicin resistant S . aureus (MRGRSA) was isolated . The clinical, microbiological and epidemiological analysis demonstrated the staphylococcal origin of the enterocolitis . The statistical analysis brought to light a significant increase in the acquisition of the infection in relation to age, surgical intervention, catheter, ulcers and antibiotic therapy . The medical staff, colonized at hand and nose by the epidemic strain contributed probably to the transmission from person to person . Antimicrobial therapy with oral vancomycin of colonized patients and application of topical ointment (betadine) on personnel eliminated colonization with methicillin/gentamicin resistant S . aureus.

Nucleic Acids Res, 1983 Nov 25, 11(22), 7679 - 93
Nucleotide sequence of the staphylokinase gene from Staphylococcus aureus; Sako T et al.; We have determined the entire nucleotide sequence of a 1,4-kilobase segment containing the staphylokinase gene, sak, molecularly cloned from the bacteriophage S phi-C genome of Staphylococcus aureus . The probable coding region is 489 base pairs long and these base pairs are translated into a polypeptide of 163 amino acid residues (Mr = 18,490) with a presumed signal sequence of 27 amino acid residues at the NH2-terminal end . In regions adjacent to the sak structural gene a possible promoter sequence and three possible terminator sequences for transcription were found about 100 base pairs upstream from the initiation codon and about 300, 400, and 500 base pairs downstream from the termination codon, respectively; they are active in an in vitro transcription system using Escherichia coli RNA polymerase . The immunoactive 18,500-dalton and 15,500-dalton proteins corresponding to a precursor form before secretion and a mature form after secretion of the sak gene products, respectively, were identified by the E . coli maxicell system.

J Biol Chem, 1983 Nov 10, 258(21), 13120 - 6
Pre-steady state beta-lactamase kinetics . The trapping of a covalent intermediate and the interpretation of pH rate profiles; Anderson EG et al.; The hydrolysis of sodium 3-dansylamidomethyl-7-beta (thienyl-2')-acetamido-ceph-3-em-4-oate, catalyzed by the beta-lactamase of Staphylococcus aureus PC1, has previously been shown (Anderson, E . G., and Pratt, R . F . (1981) J . Biol . Chem . 256, 11401-11404) to follow the reaction scheme Formula; see text . where ES' is an enzyme-substrate complex in which the substrate has undergone nucleophilic attack at the beta-lactam carbonyl group and P is product . Acid quenching of the reaction mixture has now been shown to yield, in amounts predicted by the rate constants, a covalent enzyme-substrate complex . The liability of this complex in alkaline solution is suggestive of that of an ester . Together, all of these results prove that the turnover of this apparently normal substrate by a class A beta-lactamase involves an acyl-enzyme intermediate . In the case of another fluorescent substrate, dansylcephalexin, no intermediate analogous to ES' accumulated during catalysis; presumably here, acylation of the enzyme is rate-determining . The pH profiles (pH 4-9) of the pre-steady state rate constants for hydrolysis of the former substrate have also been determined . Binding (1/K8) is pH invariant except at low pH where it weakens, probably because of substrate protonation and/or a protein conformational change . The rate constants, k2, k-2, and k3, are pH invariant at low pH but decrease at higher pH in a way which can be described by ionization of an essential acid of pKa around 7.7 . This may be the same acid for each constant, being either an active participant at the active site, or a more distant acid which controls an essential conformational change.

J Biol Chem, 1983 Nov 10, 258(21), 13262 - 7
Isolation and characterization of a 115,000-dalton matrix-associated glycoprotein from chick aorta; Bressan GM et al.; Chick aortas were extracted sequentially with phosphate-buffered saline, 6 M guanidine HCl, and 6 M guanidine HCl containing dithioerythritol . The proteins present in the guanidine HCl + dithioerythritol extract were separated by DEAE-cellulose chromatography, and the fractions recovered were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Five major glycoprotein components with apparent Mr = 205,000, 195,000, 150,000, 135,000, and 115,000 (gp 115) were identified . gp 115 was further studied since it was the only noncollagenous protein based on amino acid analysis . The protein was purified to homogeneity by preparative electrophoresis . Its amino acid composition was characterized by a high content of glutamic acid and arginine and a relatively high content of leucine, glycine, and alanine . The concentration of gp 115 in the guanidine HCl + dithioerythritol extract was about 15-fold that in the guanidine and saline extracts . Overall, about 80% of the protein was solubilized with guanidine HCl + dithioerythritol, suggesting that most of it formed large aggregates stabilized by disulfide bonds in vivo . Immunofluorescence studies with specific antibodies showed that gp 115 formed an extracellular fibrillar network in the aorta wall . One-dimensional finger printing with Staphylococcus aureus V8 protease and immunological studies indicated that the protein was unrelated to fibronectin and laminin . The data led us to conclude that gp 115 is a novel extracellular component of chick aorta.

Anal Biochem, 1983 Nov, 135(1), 221 - 9
Functional heterogeneity of monoclonal antibodies obtained using different screening assays; Mierendorf RC Jr et al.; Two alternate screening methods have enabled the detection of monoclonal antibodies with different specificities toward the lysosomal enzyme alpha-mannosidase of Dictyostelium discoideum . Spleen/myeloma hybrid cell cultures were screened for antibody production by separate assays: an indirect enzyme-linked immunoadsorbent assay (ELISA) based on the antibody binding to enzyme adsorbed on plastic, and a direct assay of the antibodies' ability to precipitate enzyme activity with fixed Staphylococcus aureus cells (Pansorbin) . Fourteen stable antibody-producing cell lines resulted from a single fusion; these fell into three distinct classes based on their screening characteristics . A group of eight were positive in both assays, and these immunoprecipitated a 140,000 Mr precursor form of alpha-mannosidase in addition to the 58,000 and 60,000 Mr mature enzyme subunits from {35S}methionine-labeled total secreted protein preparations . Two of the antibodies were positive only in the immunoprecipitation assay; these failed to precipitate the 140,000 Mr precursor . The third class consisted of four antibodies that were positive only in the ELISA method . These exclusively recognized an altered conformation of the enzyme (precursor and mature forms) that was immobilized either on plastic or on nitrocellulose paper . In addition, only members of this class were able to bind to immobilized fragments of protease-treated enzyme . The implications of these findings for the general design of monoclonal antibody screenings and for the alternative structures of this enzyme are discussed.

Allergol Immunopathol (Madr), 1983 Nov-Dec, 11(6), 457 - 64
Intrinsic polymorphonuclear chemotactic defect in a boy with chronic granulomatous disease; de la Cruz R et al.; A six year old boy is described who suffured from recurrent and protracted infections of multiple organs by various catalase positive bacteria . A severe episode of osteomyelitis involving several bones was caused by Aspergillus fumigatus . Studies of his PMNs revealed impaired metabolic as well as microbicidal functions characteristic of CGD . Chemiluminescence in response to both opsonized zymosan and sodium fluoride was markedly depressed, while control PMNs showed significant responses . Control leukocytes suspended in patient's serum likewise evoked normal chemiluminescence . Microbicidal activity against staphylococcus aureus 502A was also decreased using patient's PMNs, whereas control PMNs were able to reduce the number of colony forming bacteria by 2 logs in 120 minutes . Viable intracellular bacteria after lysis of extracellular bacteria formed 3 X 10(7) colonies from patient's PMNs and less than 2 X 10(5) colonies from the control . NBT dye reduction studies of the family members suggested an x-linked recessive mode of inheritance . The extraordinary nature of this case lies in the discovery of an associated intrinsic cellular defect of chemotaxis involving his polymorphonuclear leukocytes . Specifically, the Rebuck skin window showed predominantly mononuclear cells from 4 up to 24 hours . In addition, the patient's PMNs failed to migrate in response to cultured filtrates of E . coli as the chemoattractant . This abnormality persisted in the presence of autologous plasma or serum as well as in control plasma or serum . Control PMNs showed normal chemotaxis in the presence of the patient's plasma or serum . The extent to which the rare coexistence of these two phenomena influence the clinical disease is not known and remains to be elucidated.

Cytometry, 1983 Nov, 4(3), 254 - 62
Simultaneous measurement of phagocytosis and phagosomal pH by flow cytometry: role of polymorphonuclear neutrophilic leukocyte granules in phagosome acidification; Bassoe CF et al.; Human polymorphonuclear neutrophilic leukocytes (PMNLs) phagocytosed fluorescein-isothiocyanate (FITC)-labelled Staphylococcus aureus . Free bacteria, phagocytes, and nonphagocytes were discriminated and quantified by flow cytometry (FCM) . The relative fluorescence of phagocyte-associated and free bacteria (Nf:N) was calculated by dividing the mean phagocyte fluorescence by that of the free bacteria and the number of phagocytosed bacteria . Bactericidal capacity and chemiluminescence were measured by standard methods . The red-to-green fluorescence ratio of acridine orange-stained PMNLs (R/G) was measured by FCM . Degradation of bacteria was monitored by the reduction in FITC and ethidiumbromide fluorescence of bacteria liberated from the phagocytes . Bacterial FITC fluorescence was pH dependent . Nf:N was 0.5 to 0.7 . Using a standard curve for the interrelationship between bacterial fluorescence and pH, phagosomal pH was 5.0-5.5 . Phagocytes, kept at 4 degrees C for 24 h had Nf:N approximately 1, did not degrade bacteria, but killed them and emitted chemiluminescence . NH4Cl increased phagocyte fluorescence by 27% and decreased R/G by 50% . Cyanide and azide did not affect Nf:N . Nf:N of phagocytes from a patient with chronic granulomatous disease was 32% below, and R/G was 32% higher than the controls . Acidification of the phagosomes seems to be related to discharge of PMNL granule contents and independent of the respiratory burst.

Antimicrob Agents Chemother, 1983 Nov, 24(5), 823 - 6
Subinhibitory concentrations of antibiotics alter fibronectin binding to Staphylococcus aureus; Proctor RA et al.; Fibronectin, a high-molecular-weight glycoprotein, is found in plasma and on mammalian cell surfaces . Recent reports have suggested that bacterial-fibronectin interactions play a role in bacterial attachment to host cells . Subinhibitory concentrations of lincosamines, erythromycin, and chloramphenicol decreased fibronectin binding to Staphylococcus aureus, whereas beta-lactam antibiotics enhanced this interaction.

Antimicrob Agents Chemother, 1983 Nov, 24(5), 653 - 7
Value of serum tests in combined drug therapy of endocarditis; Drake TA et al.; Two in vitro tests, the serum killing level and the serum bactericidal rate assays, were evaluated for correlation with therapeutic efficacy in the rabbit model of Staphylococcus aureus endocarditis . Animals were treated with nafcillin alone and in combination with tobramycin or gentamicin . Both were effective therapies, but rapidity of vegetation sterilization by the single and combined regimens was shown by the serum bactericidal rate assay but not the serum killing level assay . As a direct measure of bactericidal activity in serum during therapy, the serum bactericidal rate assay may be a clinically useful supplemental test for providing information that the serum killing level assay cannot.

Plasmid, 1983 Nov, 10(3), 293 - 5
The "erythromycin-resistance" methylated sequence of Staphylococcus aureus ribosomal RNA; Ranzini AC et al.; We have determined the sequence of an oligonucleotide from the large ribosomal subunit RNA of Staphylococcus aureus whose methylation renders the organism resistant to erythromycin and other antibiotics (the "MLS" phenotype) . Analysis of RNase A digests of {3H}methyl-, 32P-labeled RNA yielded the sequence GG . m6(2)A . AAGACp, where m6(2)A is an N6-dimethylated adenosine residue that in sensitive cells is unmethylated . Comparison with homologous sequences recently reported for Saccharomyces cerevesiae mitochondria indicates that an A to G mutation in this latter system mimics dimethylation in St . aureus with regard to functional consequences.

Plasmid, 1983 Nov, 10(3), 251 - 9
Complete nucleotide sequence of pT181, a tetracycline-resistance plasmid from Staphylococcus aureus; Khan SA et al.; pT181 is a naturally occurring Staphylococcus aureus plasmid, encoding inducible resistance to tetracycline . The plasmid has a copy number of about 20 per cell, and belongs to the incompatibility group inc3 . The complete nucleotide sequence of pT181 has been determined and consists of 4437 bp . The nucleotide sequence contains 69.8% A-T and 30.2% G-C pairs . pT181 was found to contain four open reading frames capable of coding for polypeptides containing more than 50 amino acids . All the putative polypeptides are coded by one strand . The molecular weights of the four putative polypeptides are (in daltons): A, 37,500; B, 35,000; C, 23,000, and D, 18,000 . Polypeptide A corresponds to the repC protein, earlier shown to be specifically required for pT181 replication . Polypeptide B (and possibly polypeptide D) are involved in tetracycline resistance . No role has yet been established for polypeptide C; deletion of the coding sequence for the C polypeptide has no detectable effect on any property of the pT181 plasmid . A region consisting of about 1200 bp contains information for the replication and copy number control of this plasmid . The sequencing results are discussed in relation to the replication properties and tetracycline resistance associated with the pT181 plasmid.

Transfusion, 1983 Nov-Dec, 23(6), 508 - 11
Prolonged cryopreservation of human granulocytes; Richman CM; Normal human granulocytes prepared by dextran sedimentation were cryopreserved in 10 percent dimethyl sulfoxide and 25 percent autologous plasma using controlled-rate freezing at -1 degrees C per minute . Twenty-four samples were stored from 0 to 8 months in the vapor or liquid phase of liquid nitrogen . The mean cell recovery was 58 +/- 4 percent and the mean bactericidal activity using Staphylococcus aureus was 72 +/- 4 percent . Cells stored for approximately 5 months examined with transmission electron microscopy had intact cell membranes and granules although some nuclear changes were observed . No decline in cell recovery or bactericidal activity was observed with prolonged storage and there was no advantage of liquid over vapor phase . Samples stored for over 8 months showed a 73 percent cell recovery and a 77 percent bactericidal activity . Maintenance of granulocyte function after prolonged cryopreservation in these studies suggests the feasibility of cryopreserved granulocyte transfusion therapy.

J Clin Microbiol, 1983 Nov, 18(5), 1055 - 60
Serum antibodies to enterotoxins produced by Staphylococcus aureus with special reference to enterotoxin F and toxic shock syndrome; Notermans S et al.; The presence of antibodies to staphylococcal enterotoxins (enterotoxins A through F) in sera of healthy subjects (n = 567) and in sera of patients with toxic shock syndrome (n = 20) was determined . Furthermore, production of enterotoxins by Staphylococcus aureus isolated from humans was investigated . In 46, 86, 78, 41, 20, and 91% of the sera of healthy subjects, antibodies were found against enterotoxins A, B, C, D, E, and F, respectively . The high percentage of sera with antibodies against enterotoxin F correlated with the relatively high frequency of enterotoxin F-producing S . aureus isolated from humans (one-third of the isolates produced enterotoxin F) . In patients with toxic shock syndrome, antibodies against enterotoxin F were not present or were present only at very low levels . An increase of antibodies after onset of the disease was observed in two of eight patients investigated . From the results, it can be concluded that only those humans who show low levels of antibodies are susceptible to toxic shock syndrome.

Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1983 Nov, 16(4), 300 - 6
Production of penicillinases by certain penicillin-resistant bacteria . (C) factors affecting the activities of penicillinases produced by Staphylococcus aureus, S9; Elwan SH et al.; The activities of penicillinases produced by Staphylococcus aureus, S9 were found to be affected by pH, temperature, substrate concentration and type of penicillin derivative used as a substrate . The optimal activities of penicillinases produced by S . aureus, S9 were obtained at pH 6, at 37 degrees C, 0.5-10 mu/ml penicillin G concentration, and by increasing the enzyme concentration.

Infection, 1983 Nov-Dec, 11(6), 322 - 5
Susceptibility and tolerance of beta-lactamase-producing, methicillin-sensitive strains of Staphylococcus aureus towards seven broad-spectrum penicillins; Wehrli R et al.; The activity of penicillin G, ampicillin, carbenicillin, ticarcillin, azlocillin, mezlocillin and piperacillin against 102 beta-lactamase-producing, methicillin-sensitive strains of Staphylococcus aureus was determined by agar dilution (method A) and broth microdilution (method B) techniques . By NCCLS breakpoint criteria, 4% of the strains were "sensitive" to penicillin and ampicillin, and almost 100% were "sensitive" to the other drugs when method A was used . Results with method B were only significantly lower as far as the cumulative percentage of strains "sensitive" to azlocillin, mezlocillin and piperacillin was concerned (63-71%) . Bactericidal effects at "sensitive" levels were observed in 0-2% (penicillin, ampicillin), 31-35% (carbenicillin, ticarcillin) and 10-14% (azlocillin, mezlocillin, piperacillin) . While differences in MIC and MBC levels ranged from 0 to 8 dilution steps, tolerance (a greater than 32-fold difference) was seen in at least 9-22% of all strains (depending on the drug tested); experimental limitations, however, excluded a determination of tolerance in all our strains . In a semi-quantitative nitrocefin assay, "strong" beta-lactamase production was correlated to high MIC and/or MBC levels.

J Clin Microbiol, 1983 Nov, 18(5), 1226 - 36
Interlaboratory variation of antibiograms of methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains with conventional and commercial testing systems; Aldridge KE et al.; Laboratory-prepared (conventional) and commercial susceptibility testing systems were compared by using a group of methicillin-resistant (MR) and methicillin-susceptible (MS) strains of Staphylococcus aureus . A group of 25 MR and 15 MS S . aureus strains were coded and tested blindly by disk diffusion, agar dilution, broth microdilution, Sensititre, Micro-Media, Sceptor, API 3600S, MicroScan, Autobac I, and MS-2 systems . All systems were incubated at 35 degrees C and read with either a manual or automated reader at the recommended times . Where applicable, systems were also read at 48 h . Among the conventional assays, the broth and agar dilution methods were comparable, both detecting 88% of the MR strains at 24 h and detecting 92 and 96%, respectively, at 48 h . The disk diffusion method was less efficient, detecting only 36 and 72% at 24 and 48 h, respectively . Detection of cephalothin resistance was low for all systems at both time periods, with agar dilution and disk diffusion being the most and least efficient, respectively . Some variability was also seen with detection of resistance to clindamycin and gentamicin . Among the MS strains, variability among the conventional systems occurred with methicillin, gentamicin, ampicillin, and penicillin . Comparison of the commercial systems with manual readers with the broth microdilution method (reference method) showed that for MR strains, the Sceptor system gave identical results at 24 and 48 h . Sensititre detected 68 and 88% of the MR strains, whereas Micro-Media was least effective detecting 12 and 80% at 24 and 48 h, respectively . None of the commercial systems detected cephalothin resistance well, with only one strain being indicated by the Sceptor and Sensititre systems at 48 h . Slight differences were also seen among the systems with clindamycin and gentamicin . With regard to the MS strains, variability among the systems was seen with methicillin, penicillin, ampicillin, clindamycin, and gentamicin . Among commercial systems with automated readers, the API system detected a greater number of MR strains than did the reference method at 24 and 48 h, 96 and 100%, respectively . The MicroScan method was comparable to the reference method detecting 80 and 88% of the MR strains at both time periods, respectively . Both Autobac I and MS-2 were much less effective in detecting MR strains, noting only 32 and 16%, respectively, at the 3- to 6-h readings . Poor detection of cephalothin resistance among MR strains was evident in all systems . Variability also occurred among the systems with clindamycin, gentamicin, and ampicillin . A single strain of the MR group was reported to be vancomycin resistant by the API system . Among the MS group, the greatest variability was seen with methicillin . Less variability occurred with penicillin, ampicillin, gentamicin, and vancomycin.

J Immunol, 1983 Nov, 131(5), 2279 - 81
Monocyte-independent stimulation of human B lymphocytes by phorbol myristate acetate; Bertoglio JH; Phorbol myristate acetate (PMA) induces a low level of proliferation in purified human B lymphocytes when added in nanomolar concentrations to the culture medium . Much higher levels of thymidine incorporation, however, are obtained in the presence of other B cell stimuli such as anti-IgM antibodies or Staphylococcus aureus Cowan Strain 1 (SAC) . The peak activity of PMA was observed on day 3 of B cell cultures containing either anti-IgM or SAC . The rigorous depletion of monocytes as well as add-back experiments indicate that the effect of PMA on anti-IgM-stimulated B cells is not mediated by the stimulation of accessory cells . Thus, PMA acts as a very potent mitogenic agent for human B cells under culture conditions that are commonly used to assess B cell growth factor activity.

Clin Nucl Med, 1983 Nov, 8(11), 543 - 5
Gallium-67 citrate imaging in subcutaneous abscess; Sagar VV et al.; The value of Ga-67 images in identifying unsuspected locations of subcutaneous Staphylococcus aureus abscess is presented . Many of the lesions detected on the Ga-67 study were not clinically evident . In addition, follow-up studies show resolution of the changes after antibiotic therapy.

Am J Clin Nutr, 1983 Nov, 38(5), 769 - 74
Selenium deficiency with total parenteral nutrition: reversal of biochemical and functional abnormalities by selenium supplementation: a case report; Baker SS et al.; A patient with multiple intestinal fistulae maintained on total parenteral nutrition for 18 months developed low serum selenium . Erythrocyte glutathione peroxidase activity was 6% of normal . Erythrocytes were not able to metabolize H2O2 as well as those from controls, although the hexose monophosphate shunt itself was intact . Granulocytes from this patient had 15% of the erythrocyte glutathione peroxidase activity found in normals . Patient granulocytes were not able to metabolize H2O2 as well as controls, although the hexose monophosphate shunt was intact . Erythrocyte glutathione peroxidase-deficient granulocytes incubated with a respiratory burst stimulant, phorbol myristate acetate, had only 60% of the hexose monophosphate shunt activity present in control granulocytes . These abnormalities were reversed with selenium supplementation . Bacterial killing of Staphylococcus aureus 502A and cardiac function were not affected by selenium deficiency . Thus, selenium deficiency resulted in biochemical and functional abnormalities of erythrocytes and granulocytes . These abnormalities were reversed with selenium supplementation.

Surgery, 1983 Nov, 94(5), 765 - 9
A passive system using rifampin to create an infection-resistant vascular prosthesis; Powell TW et al.; Vascular prosthesis infection is a devastating complication of vascular operations . The development of a simple process for imparting infection resistance to vascular prosthesis material would be invaluable . Rifampin was added to the blood that was used to preclot 8 mm Dacron vascular prostheses that were used to replace the infrarenal aorta in 10 mongrel dogs . Rings of the grafts were resected after blood had flowed through them for 0 minute, 15 minutes, 60 minutes, and 24 hours . The resected graft rings were placed on culture plates that had been inoculated heavily with Staphylococcus aureus . The rate of antibiotic dialysis from the graft was determined by comparison of the inhibition rings that were produced by the graft rings at each subsequent interval . At 60 minutes, inhibition was 94% of that at time 0 . Inhibition at 24 hours was 91%, which demonstrated no significant decrease from 60 minutes . Other antibiotics were screened by this technique, but none of them demonstrated inhibition at 24 hours . In a pilot study in which two dogs were given parenteral rifampin before and after operation and grafts were preclotted with blood that contained rifampin, it was suggested that there was a slight increase in inhibition at 24 hours (93%) . The data indicated that rifampin that is added to the blood that is used to preclot a porous Dacron prosthesis is so slowly dialyzed from the graft that inhibition remains at 24 hours . This passive system imparts potential resistance to the prosthesis.

Rev Infect Dis, 1983 Nov-Dec, 5(6), 1003 - 11
Pyogenic psoas abscesses: noninvasive diagnostic techniques and review of the literature; Gordin F et al.; Psoas muscle abscesses are a diagnostic and therapeutic challenge . Until recently, surgery was mandated for diagnosis and drainage of these deep posterior lesions . Scanning techniques such as computerized tomography, radionuclide imaging, and ultrasonography now enable noninvasive visualization of abnormalities of the psoas muscle . Patients with abscesses in the greater psoas muscle fall into two distinct groups . Six of 12 patients reviewed had no apparent predisposing conditions . These patients presented with subacute symptoms of fever, pain, and disability . Staphylococcus aureus was the predominant organism isolated . Psoas infections developed in six other patients secondarily to infection or trauma elsewhere in the abdomen . Gram-negative and enteric organisms were the predominant bacteria isolated from this group . Surgical drainage in selected patients and appropriate antimicrobial therapy is necessary for treatment of these infections . Late complications such as osteomyelitis are not unusual.

Clin Exp Immunol, 1983 Nov, 54(2), 580 - 6
Characterization of immunological depression in mice exposed to normobaric oxygen; Levacher-Place M et al.; Immunological cell functions were evaluated during 24, 48 and 96 h O2 exposure in C57Bl/6 mice . A normobaric O2 exposure resulted in depression of delayed type hypersensitivity (DTH) to oxazolone and Staphylococcus aureus antigens . This effect was proportional to the duration of O2 exposure . The antibody response of splenic cells was more rapidly (24 h O2 exposure) and markedly depressed using a T-dependent antigen (sheep red blood cell, SRBC) than with a T-independent antigen (trinitrophenylated lipopolysaccharide, TNP-LPS) . While mitogen-induced proliferative responses of spleen cells to Con A and PHA were inhibited after 72 h of O2 exposure, proliferative responses to LPS were inhibited after 96 h . A dissociated antigen and mitogen responses was observed after a short time of O2 exposure (48 h): the antigen specific responses were impaired with a more pronounced effect on T lymphocytes, whereas the DNA synthesis in response to mitogen remained normal.

Am J Kidney Dis, 1983 Nov, 3(3), 205 - 8
Antibiotic activity in peritoneal dialysate; Rubin J et al.; There are few studies investigating whether antibiotics added to 30% glucose concentrate preserve their activity in the delivered dialysate . Using a Drake-Willock proportioning system, samples were obtained from the "to" patient path at ten minutes after starting and at four hours . Samples were tested for minimal inhibitory dilution (MID) against Escherichia coli and Staphylococcus aureus . Antibiotics evaluated included amikacin, tobramycin, gentamicin, cephalothin, cefamandole, moxalactam, ampicillin, penicillin, carbenicillin, and vancomycin . In all antibiotics studied, similar MIDs were obtained at the ten-minute and four-hour samples . Compared to saline, dialysate significantly impaired the antibiotic activity (a difference of two or more tube dilutions) of all antimicrobial agents except amikacin and vancomycin.

J Infect Dis, 1983 Nov, 148(5), 861 - 7
Resistance of Nocardia asteroides to oxygen-dependent killing by neutrophils; Filice GA; Nocardia asteroides resists killing by neutrophils despite the occurrence of the oxidative metabolic burst when the organism is phagocytosed . In a study of the apparent resistance of N asteroides to oxygen-dependent killing by neutrophils, this organism and (for comparison) Staphylococcus aureus were exposed to metabolites of the oxidative metabolic burst . N asteroides was more resistant than S aureus to H2O2, hydroxyl radical, and singlet oxygen and to the combination of H2O2, lactoperoxidase, and iodide . The rate of iodination of N asteroides by neutrophils or by the combination of lactoperoxidase and H2O2 was significantly lower than that of S aureus . Lysates of N asteroides had 2.8 times more catalase than lysates of S aureus, but levels of superoxide dismutase were similar in the two lysates . A reduction in the level of catalase activity of N asteroides with aminotriazole or azide resulted in a modest decrease in resistance to oxidative metabolites . Thus, the relative resistance of N asteroides to killing appeared to be due partially but not completely to its relatively high level of catalase activity.

Plasmid, 1983 Nov, 10(3), 270 - 8
Construction and characterization of plasmid vectors for cloning in Staphylococcus aureus and Staphylococcus carnosus; Keller G et al.; Several plasmid vectors for cloning in Staphylococcus aureus and S . carnosus have been constructed and characterized . The chimeric plasmids are composed of parts of the following parental plasmids: The chloramphenicol-resistance plasmid, pC194, the tetracycline-resistance plasmid, pMK148, and the erythromycin-resistance plasmid, pE12 . All the chimeric plasmids confer two selectable antibiotic-resistance markers on host cells . Insertional inactivation of the various antibiotic-resistance markers occurred at the BclI site of pE12, and the Sau96- or AvaII-site of pMK148; only a slight inactivation of the chloramphenicol-resistance marker occurred at the HaeIII-site of pC194 . The chimeric plasmids pCT20 and pCE10 are both stable in S . aureus and S . carnosus . In addition, the hybrid plasmids of pCT20 and pCE10, containing lambda-DNA fragments in various restriction sites between 0.4 and 1.2 kb, are stably maintained . The inserted lambda-DNA fragments appear unchanged.

Plasmid, 1983 Nov, 10(3), 260 - 9
Inhibition of Tn554 transposition: deletion analysis; Murphy E; Tn554, a transposon in Staphylococcus aureus that specifies resistance to erythromycin and spectinomycin, exhibits a high preference for a single chromosomal insertion site . If this site is already occupied by a copy of Tn554, the transposition of a second element is inhibited 100- to 1000-fold . This report defines the locus of the inhibitory activity and presents both a functional and a restriction map of Tn554 . Fragments containing parts of Tn554 were cloned on an autonomously replicating plasmid . Those clones containing the "left" end of Tn554 strongly inhibited the transposition of an incoming, intact copy of Tn554 . Analysis of deleted derivatives of these clones defined a locus tnpI, which is both necessary and sufficient for transpositional inhibition . This locus consists of the terminal 89 bp of the "left" end of Tn554 . It is suggested that this terminal sequence acts to titrate a factor required for transposition.

J Immunol, 1983 Nov, 131(5), 2273 - 8
Human-human B cell hybridomas from in vitro stimulated lymphocytes of patients with common variable immunodeficiency; Denis KA et al.; Human-human B cell hybridomas have been established from the peripheral blood lymphocytes of patients with common variable immunodeficiency (CVI) by fusion with an HGPRT-negative B lymphoblastoid cell line . IgM-secreting hybridomas were successfully obtained from CVI lymphocytes after stimulation for 5 days in vitro with a combination of PWM and Staphylococcus aureus strain Cowan I . Fusion of peripheral blood lymphocytes that were stimulated for 5 days in vitro with a single mitogen resulted in no viable hybrids from a total of 600 X 10(6) CVI lymphocytes . The combination of PWM and Cowan I did not induce appreciable Ig secretion from the CVI lymphocytes during the 5-day course, although it did so in normal lymphocytes . After the 5-day stimulation with this mitogen combination, however, a large percentage of the original number of peripheral blood cells were recovered, and these had a fusion frequency of approximately 1 to 2 per 10(6) with the B lymphoblastoid line . Fifteen cloned IgM-secreting hybridomas have been isolated from five different CVI patients . These hybridomas are tetraploid and have been stable in culture for 6 to 12 mo . All of the hybridoma lines that were examined contain a functionally rearranged IgM heavy chain gene from the B cell parent of the CVI patients . These human-human B cell hybridoma lines will enable a more thorough characterization of the B cell defects involved in CVI at the cellular and molecular levels.

J Clin Invest, 1983 Nov, 72(5), 1639 - 49
Identification and structural characterization of two incompletely processed forms of the fourth component of human complement; Chan AC et al.; Immunoprecipitates of human C4 from EDTA-plasma were incubated with {14C}methylamine and analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and fluorography . In addition to finding label in the alpha-chains of the secreted (C4s) and predominant plasma (C4p) forms of C4, two additional molecules with apparent molecular weights of approximately 168,000 (p168) and approximately 125,000 (p125) covalently incorporated methylamine, indicating the presence of an internal thioester bond . These two molecules were present at a concentration of approximately 5% of total plasma C4 and were not immunoprecipitated by antisera to C3 or alpha 2-macroglobulin . A human hepatoma-derived cell line (HepG2), in addition to synthesizing C4s and small quantities of the polypeptide precursor of C4 (pro-C4), was found to secrete p168 and p125 at concentrations of 14 +/- 4.8 and 21 +/- 9.2% (mean +/- SD), respectively, of total secreted C4 . These molecules were not found intracellularly . Both molecules were present on reduced, but not nonreduced, SDS-polyacrylamide gels . Chido (C4B) and Rodgers' (C4A) alloantisera precipitated the C4A and C4B variants of pro-C4, p168, p125, and C4s . Both tryptic and Staphylococcus aureus V8 protease peptide analyses showed homology between p168 and the beta- and alpha-chains and between p125 and the alpha- and gamma-chains . Partial NH2-terminal sequencing revealed that the beta-chain was NH2-terminal in p168 and that the alpha-chain was NH2-terminal in p125 . Taken together, these data indicate that p168 and p125 represent uncleaved beta-alpha- and alpha-gamma-fragments of pro-C4, respectively . Thus, in most individuals, plasma C4 consists of five structurally distinct molecules, the single polypeptide precursor (pro-C4), the three-subunit secreted (C4s) and predominant plasma (C4p) forms of C4, and two incompletely processed two-subunit molecules with uncleaved beta-alpha- (p168) or uncleaved alpha-gamma (p125)-subunits . In addition, all five molecules are observed for both C4A (Rodgers) and C4B (Chido) structural genes.

J Med Microbiol, 1983 Nov, 16(4), 391 - 9
Correlation of penicillinase production with phage type and susceptibility to antibiotics and heavy metals in Staphylococcus aureus; Rosdahl VT et al.; One hundred and thirty-nine bacteraemia strains of Staphylococcus aureus, representing different combinations of phage type and susceptibility to antibiotics and to cadmium (Cd), arsenate (As) and mercury (Hg), were investigated for penicillinase production . The determination of enzyme activity in induced and uninduced conditions was performed by iodometric titration . The amount of penicillinase produced could be correlated with phage pattern . Epidemically occurring strains of the 94,96 and the 83A complexes produced the largest amount of penicillinase, whereas strains of the 52,52A,80,81 complex were weaker producers . Group-II and group-III strains produced the smallest amount . Susceptibility to antibiotics and to Cd, As and Hg could not be correlated with enzyme activity, but strains resistant to penicillin plus tetracyclines and strains resistant only to Cd did produce less enzyme than strains with other resistance patterns . The percentage mean values than strains with other resistance patterns . The percentage mean values of extracellularity of the enzyme was highest amongst strains of the 94,96 complex and of type 95 . Four strains had constitutive production, one being macro-constitutive and three micro-constitutive . All four strains represented rare combinations of the above properties but were susceptible to fusidic acid . The importance of penicillinase production by epidemically occurring strains is discussed.

J Nucl Med, 1983 Nov, 24(11), 1019 - 22
Scintigraphic detection of osteomyelitis with Tc-99m MDP and Ga-67 citrate: concise communication; Graham GD et al.; Using both Tc-99m methylene diphosphonate and gallium-67 citrate, images of the lower extremities in New Zealand white rabbits were obtained on sequential days after inoculation of tibias with Staphylococcus aureus . Gallium-67 scintigraphy was positive earlier in the course of infection than Tc-99m MDP scintigraphy . In addition to 4-hr Ga-67 scintigrams, 24-hr and 48-hr scintigrams were obtained, contributing substantially to interpretation . However, 72-hr Ga-67 scintigrams contributed little additional information.

Can J Surg, 1983 Nov, 26(6), 540 - 5
Experimental colonization of vascular grafts with Staphylococcus aureus; Goeau-Brissoniere O et al.; In an attempt to solve the problem of infection of arterial grafts, the authors designed an experimental model to reproduce, in vitro, hematogenous seeding of grafts with Staphylococcus aureus . The inoculum, containing an average of 10(7) viable bacterial cells per millilitre, was circulated through grafts of various types . Normal dog aortas were used as controls . They entrapped a mean of 8 bacteria/cm2 . The prosthetic grafts, previously placed in dogs as femorofemoral arteriovenous bypasses for 2 hours, trapped many more cells: expanded polytetrafluoroethylene, 23 cells/cm2; bovine heterograft, 607 cells/cm2 and Dacron velour, 2801 cells/cm2 . All cell counts were significantly (p less than 0.001) different from control values . Thoracoabdominal aortic bypass grafts implanted in dogs 2 months previously gave the following mean numbers of trapped bacteria: expanded polytetrafluoroethylene, 19 122 cells/cm2; bovine heterograft, 863 cells/cm2 and Dacron velour, 3500 cells/cm2 . Polytetrafluoroethylene had significantly (p less than 0.001) higher numbers of trapped bacteria than any other type of prosthesis . The bacteria were located mainly on irregular fibrin strands and on surface defects of the grafts . The addition of cefazolin during the seeding process at concentrations 10 to 25 times the minimal inhibitory concentration did not decrease the numbers of bacteria in any graft . Bacterial colonization of prosthetic arterial grafts depends on the graft material and on the duration of implantation, but this study provided no answer to the controversial question of how to prevent arterial graft infections with antimicrobial agents in patients who undergo vascular procedures that expose them to bacteremia.

J Gen Virol, 1983 Nov, 64 (Pt 11), 2357 - 65
Location of an immunizing determinant within polypeptide VP1 of type O aphthovirus; Haresnape JM et al.; VP1 is the only structural polypeptide of aphthovirus able to stimulate the production of neutralizing antibody . The region of VP1 responsible for this activity was located by testing various proteolytic fragments of VP1 for their ability to compete for virus-specific antibodies in serum raised against the intact polypeptide . No antigenic activity could be detected in VP1 fragments isolated from trypsin-treated virus . Controlled digestion revealed that trypsin cleaved VP1 in four places in a preferred order, whereas chymotrypsin cut at a maximum of two sites . The initial cuts by the two proteases were made very close to each other, and in each case resulted in a greatly reduced affinity of the VP1 fragments for virus-specific antibodies in anti-VP1 serum . In contrast, aphthovirus was resistant to Staphylococcus aureus V8 protease, and treatment of isolated VP1 with this protease generated a peptide of mol . wt . 8500 which competed efficiently with virus for antiserum to VP1 and for an absorbed antiviral serum specific for trypsin-sensitive sites . The results indicate that these antisera interact specifically with aphthovirus at a single antigenic determinant located on VP1 approximately two-thirds of the way along the polypeptide sequence.

J Antibiot (Tokyo), 1983 Nov, 36(11), 1549 - 60
MIC values do not predict the intraphagocytic killing of Staphylococcus aureus by naphthalenic ansamycins; Marshall VP et al.; Ten naphthalenic ansamycins were compared for their ability to kill extracellular or phagocytosed Staphylococcus aureus 502A . These included rifamycins, streptovaricins and tolypomycin Y . Although the compounds differed markedly in killing extracellular S . aureus, there was surprisingly little difference between them in assisting human leukocytes to kill phagocytosed S . aureus . In fact, when compared to rifampin, some ansamycins that were less effective in killing extracellular bacteria were more effective in killing phagocytosed bacteria . These data, together with an analysis of structure and activity, suggested that a specific transport mechanism might be involved . First considered was a vitamin K transport mechanism . Indeed warfarin, a vitamin K antagonist, blocked the ability of rifampin to kill phagocytosed S . aureus, as did the coumarins, novobiocin and coumarin-3-carboxylic acid . However, direct evidence for a vitamin K transport mechanism could not be obtained using vitamin K preparations . The fused phenolic, bicyclic system common to all of these ansamycins was tentatively considered to be the portion necessary for phagocyte penetration.

Biochim Biophys Acta, 1983 Oct 28, 748(2), 205 - 12
Proteolysis of rat IgG subclasses by Staphylococcus aureus V8 proteinase; Rousseaux J et al.; Monoclonal IgG belonging to the four rat IgG subclasses (IgG1, IgG2a, IgG2b, IgG2c) and some IgG subclasses from normal rat serum were subjected to enzymatic degradation with Staphylococcus aureus V8 proteinase . The results show that only one subclass, IgG2b, is significantly cleaved by the enzyme, with the release of two main products identified as F(ab)2 and Fc-like fragments . This unique susceptibility of the IgG2b subclass represents therefore an easy means of identification and also offers a simple procedure for a preparation of F(ab)2 fragments from monoclonal IgG2b antibodies.

J Biol Chem, 1983 Oct 25, 258(20), 12728 - 32
Rapid purification and characterization of DNA topoisomerase I from cultured mouse mammary carcinoma FM3A cells; Ishii K et al.; We have previously shown that a DNA topoisomerase I from mouse mammary carcinoma cells is inhibited by heparin . Taking advantage of this enzyme-heparin interaction, we developed a rapid and efficient method of purification of this enzyme to near homogeneity by extraction of chromatin with 0.15 M phosphate buffer followed by two-step column chromatography on heparin-Sepharose and phenyl-Sepharose . Electrophoresis on sodium dodecyl sulfate-polyacrylamide gels revealed that the final preparation is composed of two polypeptides with apparent Mr approximately 98,000 (p98) and 102,000 (p102), p98 comprising 70% and p102 30% . Extraction and renaturation of the polypeptides from the gel shows that both p98 and p102 seem to possess topoisomerase activity . Partial proteolytic digestion of p98 and p102 with Staphylococcus aureus V8 and chymotrypsin yielded a series of identical peptides, indicating that the two polypeptides are structurally related . The enzyme sedimented through sucrose density gradient with s20,w of 4.0 S, and thus is monomeric in solution.

J Biol Chem, 1983 Oct 25, 258(20), 12702 - 6
Site of action of a ribosomal RNA methylase responsible for resistance to erythromycin and other antibiotics; Skinner R et al.; The enzyme which confers resistance to erythromycin in the producing organism Streptomyces erythraeus dimethylates a single adenine residue in Bacillus stearothermophilus 23 S rRNA . This corresponds to residue Ade 2058 in Escherichia coli 23 S RNA . The methylase responsible for resistance to macrolides, lincomycin, and streptogramin B-related antibiotics in Staphylococcus aureus also acts at this site.

Eur J Biochem, 1983 Oct 17, 136(1), 89 - 99
Covalent structure of chicken pepsinogen; Baudys M et al.; Chicken pepsinogen is a glycoprotein consisting of a single polypeptide chain and containing the following 367 amino acid residues: Asp23, Asn16, Thr26, Ser41, Glu14, Gln11, Pro18, Gly31, Ala17, Cys7, Val25, Met9, Ile23, Leu28, Tyr22, Phe20, His8, Lys17, Arg7, Trp4 . The Mr-value of the protein is 42 074 . This value includes the carbohydrate moiety of the protein, i.e . Man3, (GlcNAc)7, (-SO3H)5 . The primary fragmentation of the molecule was effected by limited trypsinolysis at arginine residues after preceding modification of the lysines with citraconic anhydride . All eight peptides expected in theory were obtained and their size, amino acid composition, and N-terminal amino acid sequence were characterized . To elucidate the amino acid sequence of these large fragments the latter were subjected to secondary cleavage by CNBr, trypsin (after removal of the protecting groups from the lysines), the proteinase from Staphylococcus aureus V8 strain, alpha-chymotrypsin, hydroxylamine, or dilute acid; the resulting peptides were isolated by gel permeation and ion-exchange chromatography and by the fingerprint techniques . Overlaps at sites of the arginine residues were obtained in an earlier study {Baudys, M . & Kostka, V . (1982) Collect . Czech . Chem . Commun . 47, 2814-2832} . Chicken pepsinogen shows the highest degree of homology with the primary structures of pepsinogens A . The internal homologies are apparent in the neighborhood of the two active aspartic acid residues . We have assigned tentatively chicken pepsinogen to the group of pepsinogens A (EC 3.4.23.1); this assignment is a result both of our sequence studies and of an investigation of the kinetic characteristics of the enzyme.

Eur J Biochem, 1983 Oct 17, 136(1), 101 - 6
Purification, subunit structure and immunological comparison of fructose-bisphosphate aldolases from spinach and corn leaves; Kruger I et al.; The cytosol and chloroplast fructose-bisphosphate aldolases from spinach leaves were separated by ion-exchange chromatography on DEAE-cellulose, and were purified by subsequent affinity chromatography on phosphocellulose to apparent homogeneity as judged from polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate . The two aldolases had specific activities of 7.2 and 7.8 units mg protein-1 . Molecular weight determinations by electrophoresis in sodium dodecyl sulfate gels and by sedimentation velocity centrifugation in sucrose gradients showed that the aldolases contained four subunits of Mr 38 000 and 35 000, respectively . Antibodies against the cytosol and chloroplast aldolase from spinach leaves were raised in a guinea pig and in a rabbit, respectively . In the Ouchterlony double-diffusion test, the two aldolases did not cross-react . A small degree of cross-reaction was observed by a test in which immune complexes were adsorbed to a solid-phase support (Staphylococcus aureus Cowan I cells) and nonbound enzyme activity was determined after centrifugation . These results imply major structural differences between the two spinach leaf aldolases . Only one major aldolase could be resolved on DEAE-cellulose from corn leaves . The aldolase was purified and had a specific activity of 6.4 units X mg protein-1 . The corn leaf aldolase cross-reacted with the antiserum raised against the chloroplast enzyme from spinach leaves, but not with the other antiserum . Thus, the corn leaf aldolase could be identified as a chloroplast enzyme . Since aldolase activity is mostly restricted to the bundle sheath cells of corn leaf, it was concluded that it is compartmentalized in the chloroplasts of these cells but not in chloroplasts of the mesophyll cells.

Arch Biochem Biophys, 1983 Oct 15, 226(2), 643 - 56
Amino acid sequence of bovine white matter proteolipid; Lees MB et al.; The sequence of the bovine white matter proteolipid has been studied by a combination of proteolytic digestion and chemical cleavage at tryptophan residues . Alignment of peptides obtained by digestion with trypsin, chymotrypsin, clostripain, and Staphylococcus aureus protease gave the sequence of 52 residues at the amino terminus, 96 residues at the carboxyl terminus, and several additional segments . Peptides obtained by treatment of the protein with 2-(2-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine confirmed the alignment and extended the sequence . This information, combined with that of other investigators, permits us to propose the primary structure for the entire protein . On the basis of the sequence determination, the molecular weight of the proteolipid protein is 29,869.

J Immunol Methods, 1983 Oct 14, 63(2), 145 - 57
Radioimmunoassays for protein A of Staphylococcus aureus; Langone JJ et al.; Radioimmunoassays have been developed that can detect nanogram amounts of protein A (SpA), a product generated by Staphylococcus aureus that binds selectively to the Fc region of IgG from most mammalian species . Competition assays for fluid phase SpA utilize antibodies produced in chickens, 125I-labeled SpA as the tracer molecule, and either F(ab')2 fragments of rabbit IgG anti-chicken IgG or 40% ammonium sulfate as the precipitating agent to separate antigen-antibody complexes from free antigen . The double antibody assay could be carried out in serum from species that form only soluble complexes with SpA (e.g., rabbit), that react poorly with SpA (e.g., rat), or under appropriate conditions in serum from species (e.g., dog) that show high reactivity with SpA and form precipitating complexes . Chicken antibodies prepared by affinity chromatography on SpA-Sepharose and labeled with 125I were used in a direct binding assay for SpA present either on the cell wall of Cowan strain I or Wood 46 bacteria, in insoluble complexes prepared from SpA and whole serum or purified IgG, or in Clq binding complexes that were formed by passage of serum from normal or tumor bearing humans or dogs over SpA-collodion charcoal . Since both types of assays could detect SpA even in the presence of serum or IgG, they offer advantages over other techniques in which the SpA-Fc interaction may interfere.

Brain Res, 1983 Oct 3, 276(1), 127 - 39
Specificity of antisera prepared against pure bovine MAO-B; Pintar JE et al.; Antisera have been prepared against purified bovine MAO-B that appear to react selectively with MAO-B and not MAO-A, Rabbit and mouse antisera indirectly immune precipitated {125I}bovine MAO-B using inactivated Staphylococcus aureus cells, and binding of antibodies to bovine and rat MAO-B did not inhibit enzyme activity . Two continuous rat cell lines, hepatoma line MH1C1 and glioma line C6, were used to elucidate the specificity of the antisera . MH1C1 cells, which express both MAO-A and MAO-B, showed immune-specific staining with rabbit antiserum, and staining was blocked with pure MAO-B . Further, MAO-B activity and {3H}pargyline-labeled MAO molecules could be immune precipitated from solubilized mitochondrial preparations of MH1C1 cells; and immune fixation of mitochondrial proteins following SDS polyacrylamide gel electrophoresis (SDS-PAGE) revealed staining of the MAO-B, but not of the MAO-A, flavin-containing subunit . In contrast, no immune-specific immunocytochemical staining was observed in C6 cells, which have only MAO-A activity; no MAO-A activity or {3H}pargyline-labeled MAO could be immune precipitated from solubilized mitochondrial preparations of these cells, and no stained bands were observed for mitochondrial proteins resolved by SDS-PAGE and processed for immune fixation . Further support for the selectivity of this antiserum for MAO-B comes from immunocytochemical staining of rat tissues which express varying amounts of MAO-A and MAO-B activities . Hypothalamus and liver, with high levels of MAO-A and MAO-B activities showed a large number of immunoreactive cells, whereas spleen, heart and superior cervical ganglia, with high MAO-A and low MAO-B activities showed only a few or no stained cells . Catecholamine neurons in the substantia nigra, thought to contain MAO-A, did not show immune-specific staining . Skeletal muscle cells with low MAO-A and MAO-B activities did not stain . These studies provide additional evidence that MAO-A and MAO-B are distinct molecules, differentially expressed in different cell types.

Pediatrics, 1983 Oct, 72(4), 476 - 80
Epidural abscess and vertebral osteomyelitis following serial lumbar punctures; Bergman I et al.; Lumbar epidural abscess and vertebral osteomyelitis were diagnosed in a 3-month-old infant, born prematurely, who had had repeated lumbar punctures for the treatment of posthemorrhagic hydrocephalus . Staphylococcus aureus was the causative organism . Successful treatment was achieved with 6 weeks of intravenous antibiotics without surgical drainage . Infectious complications of lumbar punctures are rare, but may occur when multiple punctures are attempted in small premature infants whose subarachnoid space contains large amounts of blood . Infection can be introduced directly by a contaminated spinal needle, or trauma to the tissues with bleeding can create a favorable site for bacterial adherence and multiplication . Posthemorrhagic ventricular dilation often resolves spontaneously and serial lumbar punctures should be used to treat this condition only when CSF flow is easy to establish and maintain.

J Virol, 1983 Oct, 48(1), 10 - 7
In vitro and in vivo studies of bovine parvovirus proteins; Lederman M et al.; Total cytoplasmic RNA from bovine parvovirus (BPV)-infected cells or BPV-specific RNA selected by hybridization to cloned BPV genomic sequences were translated in a message-dependent rabbit reticulocyte lysate . Immunoprecipitation, using immunoglobulin G from rabbits injected with purified BPV, resulted in the detection of {35S}methionine-labeled polypeptides with MrS of 80,000, 72,000, 62,000, and 60,000 . These in vitro translation products had the same mobility on sodium dodecyl sulfate-polyacrylamide gels as that of the four proteins found in purified virions . The three largest polypeptides had amino acid sequence homology, as judged by serological methods and partial proteolysis with Staphylococcus aureus V8 protease . Additional noncapsid proteins with MrS of 25,000, 27,000, and 31,000 were also detected as translation products of these RNAs . All of the above species were immunoprecipitated by immunoglobulin G from a calf which was naturally infected with BPV . All four capsid proteins but only one of the lower-molecular-weight polypeptides were detected after the immunoprecipitation of BPV-infected cells . The results presented here indicate that the BPV genome codes for four capsid proteins and a noncapsid protein which may be structurally related to the capsid proteins.

J Pathol, 1983 Oct, 141(2), 157 - 67
Acute haematogenous osteomyelitis: an experimental model; Emslie KR et al.; A simple and reproducible model of acute haematogenous staphylococcal osteomyelitis is described . Twenty-nine day-old chickens were inoculated intravenously with 10(4)-10(8) viable organisms Staphylococcus aureus per kg body weight and were killed 1-8 days after inoculation . Macroscopic septic foci could be detected within 24 hr of inoculation and were most commonly situated in the metaphyseal region of the proximal tibia and distal femur . Lesions in other organs were not observed . The production of osteomyelitis was dependent on the bacterial inoculum size . It was estimated that 5.5 X 10(5) viable organisms per kg body weight of chicken were required to produce osteomyelitis in 50 per cent of injected chickens . Chicken weights were monitored throughout the experiment . A close negative correlation existed between the logarithm of the bacterial inoculum size and the chicken growth rate in the first 24 hr following inoculation (r = -0.968, P less than 0.01) . The chicken growth rate was therefore used as an accurate predictor of osteomyelitis in individual chickens.

Acta Pathol Microbiol Immunol Scand {B}, 1983 Oct, 91(5), 351 - 6
Antibody response to alpha- and betahemolysin from Staphylococcus aureus in patients with staphylococcal infections and in normals; Christensson B et al.; One hundred and nineteen patients with S . aureus infections and 22 patients with non-S . aureus septicemia were investigated for anti-alpha hemolysin antibodies using a radioimmunoassay (RIA) . As compared to 16- healthy controls, patients with S . aureus endocarditis, septicemia, chronic osteomyelitis and recurrent furunculosis showed significantly higher antibody levels, while the non-S . aureus septicemia group showed normal levels . Corresponding results were obtained using the conventional anti-staphylolysin (ASTA) test . Only patients with recurrent furunculosis had significantly elevated anti-beta hemolysin antibody levels assessed by RIA, in comparison with healthy controls . The highest antibody levels were found in furunculosis patients infected with S . aureus strains which were high producers of beta hemolysin . The results indicate that furunculosis patients do not have a defective serological response against S . aureus beta hemolysin.

Antibiotiki, 1983 Oct, 28(10), 767 - 71
{Microflora of the burned, its sensitivity to antibiotics and bacteriophages}; Panchenkov NR et al.; Pathogenic strains of Staphylococcus aureus remain the most frequent causative agents of infection in patients with burns . With expansion of deep burns the frequency of gram-negative bacteria increased . The isolates were highly resistant to antibiotics . This required constant control of the sensitivity to antibiotics and bacteriophages.

J Antimicrob Chemother, 1983 Oct, 12 Suppl C, 85 - 95
Effects of subinhibitory concentrations of antibiotics on Staphylococcus aureus interactions with fibronectin; Proctor RA et al.; Bacterial adherence to host tissues relies on interactions between tissue macromolecules and bacterial surface molecules . One of the major predisposing factors to infection with Staphylococcus aureus is trauma to tissues . A common element in traumatized tissues is fibronectin . In previous studies, we have shown that fibronectin binds to Staph . aureus . In this paper, we have investigated the effects of subinhibitory concentrations of antibiotics on fibronectin interactions with Staph . aureus . Exposure of Staph . aureus to 1/4 MIC of penicillin increases the number of binding sites and enhances adherence of Staph . aureus to a collagen-fibronectin matrix . Chloramphenicol, erythromycin, clindamycin, and U57,930E all decreased the number of binding sites . Also, U57,930E reduced Staph . aureus adherence to a collagen-fibronectin matrix . Taken together, these data suggest that penicillin may enhance Staph . aureus adherence to tissue fibronectin whereas U57,930E might reduce such binding.

J Antimicrob Chemother, 1983 Oct, 12 Suppl C, 51 - 62
Interactions of Bacteroides fragilis and phagocytes: studies with whole organisms, purified capsular polysaccharide and clindamycin-treated bacteria; Wade BH et al.; Bacteroides fragilis plays a key role in the pathogenesis of anaerobic infections and is often found mixed with aerobic organisms . We explored the interactions of this organism with phagocytes in an attempt to discern additional information about its virulence factors . We confirm an earlier report that killing of aerobic organisms by polymorphonuclear leukocytes (PMN) is decreased in the presence of high numbers of Bact . fragilis but this effect could also be demonstrated with Bact . distasonis or Staphylococcus aureus . Our data support the concept that this phenomenon may be due to competition for opsonins . Virulence of Bact . fragilis has been associated with a polysaccharide capsule . We were unable to demonstrate any deleterious effect of the purified capsular polysaccharide of Bact . fragilis on phagocytosis, killing, or chemotaxis by PMN . We were not able to demonstrate any effect of subinhibitory levels of clindamycin on the interactions of neutrophils and Bact . fragilis.

Hoppe Seylers Z Physiol Chem, 1983 Oct, 364(10), 1383 - 409
Hemocyanins in spiders, XIX . Complete amino-acid sequence of subunit d from Eurypelma californicum hemocyanin, and comparison to chain e; Schartau W et al.; The complete primary structure of subunit d of the hemocyanin from the tarantula Eurypelma californicum was determined by manual micro sequencing . Subunit d of Mr = 73000 is split about in the middle of the chain during limited trypsinolysis, only one single bond being attacked . The whole chain contains 14 methionine residues and after cyanogen bromide cleavage 15 peptides could be isolated by gel and ion exchange chromatography and high pressure liquid chromatography . The cyanogen bromide peptides and the large (Mr = 34000 and 37000, respectively) fragments resulting from limited trypsinolysis, were further cleaved with trypsin, chymotrypsin, Staphylococcus aureus proteinase, formic acid, and Astacus fluviatilis proteinase, the latter being very useful in obtaining certain overlapping peptides . The total chain length is 627 residues . Carbohydrate side chains were not found . The sequence is discussed with respect to the gross physical properties of the subunit, to homologies with subunit e and the cleavage specifities of the enzymes employed.

J Infect Dis . 1983 Oct;148(4):764.
Effect of Staphylococcus aureus enterotoxin F on human neutrophil oxidative metabolism; Goetz MB et al.; Epidemiologic and animal challenge studies have suggested that SEF may play a critical role in toxic-shock syndrome {1} . However, the pathogenic mechanism of SEF activity is not known . One means by which this toxin could elicit the widespread organ dysfunction observed in toxic-shock syndrome would be by directly promoting PMN production of toxic oxygen species and the resultant secondary endothelial cell damage {2} . To evaluate this possibility, we assayed the effect of SEF on PMN oxidative metabolism . SEF at 0.01-100 ng/ml did not stimulate O2- release or O2 consumption by inactive PMNs . Similarly, incubation of PMNs with 10 ng of SEF/ml for 1 hr neither potentiated nor inhibited cellular O2 consumption stimulated by optimal (10 mg/ml) or suboptimal (0.1 mg/ml) concentrations of opsonized zymosan . Finally, SEF had no effect on O2- release by PMNs stimulated by PMA . PMN viability, as assessed by trypan blue exclusion, was unaffected by SEF . This study did not address the possibility that SEF might indirectly activate PMN oxidative metabolism by promoting leukocytic pyrogen production by monocytes and macrophages {3} . SEF neither directly activated PMN oxidative activity nor potentiated the cellular oxidative response to particulate or soluble stimuli . Consequently, direct stimulation of PMN-derived, O2- mediated damage to endothelial cells is not a tenable hypothesis to explain the mechanism of SEF toxicity.

J Infect Dis . 1983 Oct;148(4):763.
Impact of methicillin-resistant Staphylococcus aureus on the incidence of nosocomial staphylococcal infections; Boyce JM et al.; MRSA strains have become increasingly prevalent in the United States and are now an important cause of nosocomial infections in many large, medical school-affiliated hospitals . In affected institutions, from a few percent to 50% of all hospital-acquired S aureus infections are caused by MRSA strains . It has been suggested that the overall incidence of nosocomial S aureus infections may not increase in hospitals where MRSA strains have become epidemic or endemic and that MRSA strains merely replace methicillin-susceptible strains as a cause of hospital-acquired infections . Several recent studies lend support to this theory . Thompson et al {1} reported that the overall incidence of nosocomial S aureus-associated bacteremias and postoperative wound infections in a university hospital did not increase during a period when MRSA strains caused a significantly greater proportion of such infections . Similarly, Linnemann et al {2} found that the overall incidence of nosocomial S aureus-associated bacteremias did not change during a four-year period when the incidence of MRSA-associated bacteremias increased appreciably . At the University of Mississippi Medical Center, MRSA strains have been recovered from patients with increased frequency since an outbreak of MRSA infections occurred in the burn unit in June 1979 {3} . Continuing surveillance has revealed that the incidence of nosocomial MRSA infections was significantly higher in 1980-1982 than during 1979 (P = 0.002 by Mann-Whitney U test) . MRSA strains accounted for 11% of nosocomial S aureus infections in 1979, 38% in 1980, 50% in 1981, 36% in 1982, and 32% in early 1983.(ABSTRACT TRUNCATED AT 250 WORDS)

J Infect Dis, 1983 Oct, 148(4), 692 - 8
Prevalence of serum antibody to staphylococcal enterotoxin F among Wisconsin residents: implications for toxic-shock syndrome; Vergeront JM et al.; Staphylococcal enterotoxin F (SEF) has previously been shown to be a marker for toxic-shock syndrome (TSS)-associated strains of Staphylococcus aureus, whereas the serologic absence of antibody to SEF (anti-SEF) has been shown to be a marker for susceptibility of persons to TSS . In this study, anti-SEF was measured by radioimmunoassay in 689 banked sera obtained from Wisconsin residents during 1960, 1970, and 1980 . The prevalence of anti-SEF as estimated by logistic regression analysis was 47%, 58%, 70%, 88%, 96%, and 99% at ages one, five, 10, 20, 30, and 50 years, respectively . Evidence for the transplacental transfer of anti-SEF is also presented . Despite the reported increased incidence of TSS occurring during the past five years, with a preponderance of cases occurring among women, no significant differences in the prevalence of anti-SEF were noted between sexes or longitudinally between the years 1960, 1970, and 1980 . These data enhance our understanding of the epidemiology of TSS and further identify the population that may be susceptible to TSS.

Br J Urol, 1983 Oct, 55(5), 564 - 7
Wound infection following vasectomy; Randall PE et al.; PIP: 94 patients undergoing vasectomy as day cases were studied prospectively to assess the true wound infection rate for vasectomy in the Hope Hospital in Salford Manchester, England, to assess the subsequent morbidity, and to elucidate any factors that may be responsible for infection . The pilot study had already indicated an unacceptably high infection rate and so it was decided to investigate the value of a preoperative hibiscrub shower in reducing the size of the problem . All patients had nasal, scrotal, and perineal swabs taken, the swabs being taken by rolling the swab several times against the area to be sampled . The patients were then randomly assigned to 3 groups . Group 1 consisted of 32 patients undergoing vasectomy alone with no preoperative shower; Group 2, 32 patients undergoing a single preoperative hibiscrub shower; and group 3, 30 patients undergoing a single preoperative shower with ordinary soap . All patients were assessed 7 days postoperatively for wound infection and hematoma formation . The patients also were questioned as to time off work . An infected wound was considered to be any wound that was open and discharging either purulent or serous fluid . In the context of vasectomy an erythematous wound was not considered to be infected, this being part of the inflammatory reaction caused by the catgut skin closure . If the wound was infected a swab was taken . Of the 94 patients, 83 returned postoperatively . The 10 who were contacted at home reported no problems and only 1 patient was lost to follow-up . There were 31 infections among the 94 patients, an overall infection rate of 32.9% . 4 infections were severe and 3 of these had an associated epididymoorchitis . 21 patients (22%) developed hematomas under the wound . None of these was more than 1.5 cm in diameter . 9 of the infected patients had time off work because of the infection . 3 of these were severe infections but 6 were mild . 9 of the noninfected patients also had time off work, the reasons being given being swelling (4) and pain (5) . Staphylococcus aureus accounted for 60% of the infections and in only 1 case was it part of a mixed growth . All of the other infections were mixed . No perineal carriers of Staph . aureus were encountered but 15 patients (16%) were found to be nasal carriers of this organism . Phage typing revealed that only 3 of the staphylococcal wound infections were due to the same organisms as found on the nasal swab . Of the Staph . aureus wound infections, 1 group of 3 cases and a separate group of 4 cases revealed the same phage types . The most common organisms found on the scrotum and perineum preoperatively were Staphylococcus epidermidis and diptheroids . A soap shower exerted no significant effect on the number of organisms .

Radiat Res, 1983 Oct, 96(1), 41 - 50
Radiosensitization of Staphylococcus aureus by secobarbital sodium and other barbiturates; Sade N et al.; The barbiturate hypnotic, secobarbital sodium, at millimolar concentrations, sensitizes Staphylococcus aureus in anoxic buffer-saline suspension (pH 7.0) to the lethal effects of gamma rays . The maximal response represents 50% of that for oxygenated suspensions without additive . Secobarbital sodium operates within the oxygen effect . It must be present at the time of irradiation for modification of radiation response . This, coupled with its testing in the presence of other additives, points to its involvement in an intracellular reaction with a radiation-induced short-lived chemical species, probably the electron . Preliminary tests show that pentobarbital sodium also operates as an efficient hypoxic radiosensitizer . Lack of sensitization by phenobarbital sodium is attributed to its low lipid solubility.

Hoppe Seylers Z Physiol Chem, 1983 Oct, 364(10), 1347 - 56
{The primary structure of the alpha-amylase inhibitor Hoe 467A from Streptomyces tendae 4158 . A new class of inhibitors}; Aschauer H et al.; The native or modified alpha-amylase inhibitor Hoe 467A - isolated from the culture medium of Streptomyces tendae 4158 - and overlapping peptides were degraded by the automatic Edman technique . The oxidized or aminoethylated or oxidized and maleoylated inhibitor was digested with trypsin and the native inhibitor with pepsin . Further digestion with Staphylococcus aureus proteinase was also carried out . After peptic digestion two cystin peptides were isolated, which allowed the establishment of the disulfide bonds . The alpha-amylase inhibitor is a polypeptid consisting of 74 amino-acid residues with a molecular mass of 7958 Da . The inhibitor is composed of all naturally occurring amino acids except methionine and phenylalanine and shows no sequence homology to known inhibitors . The clinical and pharmacological importance in respect to the inhibitors ability for inactivation of human salivary and pancreatic alpha-amylase is discussed . Especially the proteinase resistance of the inhibitor enables a clinical application in human (e.g . Diabetes mellitus) per os.

J Antibiot (Tokyo), 1983 Oct, 36(10), 1380 - 6
Release of lipoteichoic acid from Staphylococcus aureus by treatment with cefmetazole and other beta-lactam antibiotics; Utsui Y et al.; The effect of cefmetazole on the growth together with the release of cellular lipoteichoic acid from cefazolin-resistant strains of Staphylococcus aureus was compared with that of cefazolin, cefotiam, cefoxitin and cefuroxime . Bacteriolytic actions were measured by turbidity and bactericidal actions were followed by viable cell count . Release of cellular lipoteichoic acid was measured by the radioactivity in the supernatant of the cultures . Cefmetazole exerted more potent effects on the bacterial growth and induced more marked release of cellular lipoteichoic acid from resistant strains as compared with other beta-lactams.

Microbiologica, 1983 Oct, 6(4), 277 - 91
Biochemical and physical properties of the endo-beta-N-acetylglucosaminidases from Staphylococcus aureus, Staphylococcus simulans, and Staphylococcus saprophyticus; Valisena S et al.; Biochemical and physical properties of the pure bacteriolytic enzymes excreted by three different Staphylococcus species (S . aureus, S . simulans, and S . saprophyticus) were investigated . Although the three enzymes have previously been shown to share the same specificity of action (endo-beta-N-acetylglucosaminidase activity), their biochemical features clearly indicated that they were three different enzymes, confirming what had previously been suggested by the different lytic-activity patterns displayed by each species and the different procedures needed to achieve purification of each enzyme . Very different values resulted from molecular weight determination: 80,000 for the S . aureus enzyme, 45,000 for the S . simulans enzyme and 31,000 for the S . saprophyticus enzyme . Other important differences were observed in their kinetics of activity on Micrococcus luteus purified cell walls; their stability; their bacteriolytic spectrum against heat-killed cells of various microorganisms; and their response to physical and chemical factors, such as temperature, pH, ionic strength, divalent cations, chelating agents, thiol compounds, and glucose derivatives.

J Rheumatol, 1983 Oct, 10(5), 688 - 93
Influence of agents with immunomodulating activity on phagocytosis and bactericidal function of human polymorphonuclear cells; Pruzanski W et al.; Six immunomodulators were tested for the influence on phagocytosis and intracellular bactericidal activity of human polymorphonuclear cells (PMN) . Frentizole, lamprene, intal and levamisole but not dapsone enhanced phagocytosis in the presence of serum . Without serum, frentizole strongly enhanced phagocytosis, whereas lamprene, levamisole and dapsone had weaker enhancing activity . Rifampin suppressed phagocytosis of Staphylococcus aureus whether the serum was present or not . The influence on phagocytosis was time and dose dependent . All drugs but dapsone markedly enhanced intracellular bactericidal activity of PMN in a dose dependent fashion . Dapsone enhanced bactericidal activity without the serum and suppressed it in its presence . It may be concluded that immunomodulators are a heterogeneous group of substances and their influence on phagocytosis and cellular bactericidal activity varies . Enhancing activity of some immunomodulators implies that they may be used in conditions with impaired phagocytosis.

Immunol Commun, 1983 Oct, 12(5), 453 - 64
Inhibition of rat mammary tumor growth by purified protein A--a potential anti-tumor agent; Ray PK et al.; Intravenous inoculations of purified Protein A of Staphylococcus aureus Cowan I cause significant (p less than 0.01) regression of di-methyl-benz-anthracene (DMBA)-induced mammary adenocarcinomas in Sprague-Dawley rats . Direct tumor cell counts of treated tumors showed fewer (p less than 0.005) viable tumor cells than did tumors from untreated controls . Plasma immunoglobulin G concentration showed a significant (p less than 0.05) increase compared to that of controls . However, the concentration of immune complexes and percentages of T-rosettes did not change . Peripheral blood mononuclear cells (PBMNC) from treated animals showed increased cytotoxicity (p less than 0.005) compared to that of controls . Plasma of treated animals potentiated PBMNC cytotoxicity (p less than 0.05) and showed increased antibody and complement-mediated cytotoxicity (p less than 0.025) . The exact mechanism of protein A-induced potentiation of anti-tumor immune reactivities leading to tumoricidal response is not known . However, our data are suggestive of the involvement of both cellular and humoral immunity of the host in the tumor regressive phenomenon.

J Clin Microbiol, 1983 Oct, 18(4), 895 - 900
Production of monoclonal antibodies to Legionella pneumophila serogroups 1 and 6; Para MF et al.; To better define the surface antigens of Legionella pneumophila for clinical and experimental purposes, we have produced monoclonal antibodies to L . pneumophila serogroups 1 and 6 . Two hybridomas were produced in serogroup 1 . One antibody, LP-I-17, recognized a serogroup-common antigen . The second antibody, LP-I-81, was specific for serogroup 1 . This antibody was able to agglutinate bacterial cells belonging to the serogroup 1 reference strains . Philadelphia and Knoxville . Microagglutination assays of environmental and clinical isolates revealed a subgroup of serogroup 1 environmental isolates which were not agglutinated by LP-I-81 . This subset of isolates was segregated to certain buildings in the medical complex . Immunodiffusion studies showed identity between the LP-I-81 antigen and the serogroup-specific antigen of serogroup 1 organisms . This antigen could be absorbed out of the serogroup 1 organism extract with LP-I-81-coated Staphylococcus aureus, leaving the serogroup-common antigens . Three hybridomas were produced to serogroup 6 . All three produced antibodies which were serogroup 6 specific and agglutinated serogroup 6 bacteria.

Acta Pathol Microbiol Immunol Scand {B}, 1983 Oct, 91(5), 307 - 10
Comparison of four methods for differentiation of Staphylococcus aureus from other Micrococcaceae in the routine laboratory; Dibb WL et al.; Four methods for the identification of Staphylococcus aureus (tube coagulase test, thermostable nuclease test, indirect agglutination of fibrinogen coated erythrocytes and a commercial latex kit: SeroSTAT Staphylococcus Test) have been compared . Clinical isolates (698) and 40 reference strains of Micrococcaceae were included in the study together with control organisms . The coagulase test gave no false positive results but 39/406 clinical isolates of S . aureus were negative at 2h and one half were only weakly positive . At 18 h, all but 2 of 406 isolates gave a positive reaction . The thermostable nuclease test was very specific; no clinical isolates of S . aureus gave negative results and no "coagulase-negative" clinical isolates gave a definite positive reaction . The indirect haemagglutination method was sometimes difficult to interpret and frequently gave negative or doubtful results for S . aureus . The SeroSTAT test was easy to use and interpret and was specific; the method is suitable for routine laboratory use, particularly when a rapid result is desirable.

J Antibiot (Tokyo), 1983 Oct, 36(10), 1290 - 4
Safracins, new antitumor antibiotics . III . Biological activity; Ikeda Y et al.; Safracins A and B have antibacterial activity against Gram-positive and Gram-negative bacteria in vitro but no therapeutic activity in mice infected with Staphylococcus aureus . Safracins A and B induce abnormal morphological changes in Echerichia coli cells . Tests with transplantable mice tumors demonstrate that safracins A and B inhibit the growth of P388 leukemia and IMC carcinoma.

Hoppe Seylers Z Physiol Chem, 1983 Oct, 364(10), 1357 - 81
Hemocyanins in Spiders, XVIII . Complete amino-acid sequence of subunit e from Eurypelma californicum hemocyanin; Schneider HJ et al.; The complete amino-acid sequence of subunit e of the hemocyanin from the tarantula, Eurypelma californicum, was determined by a combination of manual and automated methods . By limited proteolysis with chymotrypsin, two large fragments (e-CHn 29 and e-CHn 42) were obtained . The large peptides were further cleaved with cyanogen bromide, trypsin (with and without prior blocking of lysine residues), chymotrypsin, Staphylococcus aureus proteinase, Astacus fluviatilis proteinase, or 25% formic acid . The complete chain comprises 621 residues . A remarkable feature of the sequence is a hexapeptide -His-His-Trp-His-Trp-His- which is believed to take part in the binding of copper.

J Infect Dis, 1983 Oct, 148(4), 682 - 91
Activation of purified human plasma prekallikrein triggered by cell wall fractions of Escherichia coli and Staphylococcus aureus; Kalter ES et al.; Whether Escherichia coli and Staphylococcus aureus cell wall fractions can trigger the activation of prekallikrein was investigated in a mixture of purified human factor XII, prekallikrein, and high-relative-molecular-weight (Mr) kininogen . After exposure for 30 min to bacterial preparations (0.02-5 mg/ml) at 0 C, lallikrein amidolytic activity was expressed as a percentage of the optimal activation of prekallikrein induced by dextran sulfate . Lipopolysaccharide (LPS) fractions of five E coli strains and lipid A of E coli O111B4 induced 50%-90% optimal activity . However, the polysaccharide fraction induced less than 5% activity . Peptidoglycan and teichoic acid of S aureus induced 70%-100% optimal activity at 5 mg/ml, but protein A did not generate activity . No activation of prekallikrein occurred in the absence of factor XII . Thus, LPS and lipid A of E coli and peptidoglycan and teichoic acid of S aureus can generate kallikrein amidolytic activity in a mixture of purified factor XII, prekallikrein, and high-Mr kininogen.

Biull Eksp Biol Med, 1983 Oct, 96(10), 102 - 4
{Phagocytosis of bacteria by polymorphonuclear leukocytes suspended in liquid or adhering to a surface}; Pal'tsyn AA et al.; Phagocytosis of Staphylococcus aureus by polymorphonuclear leukocytes (PNL) was studied in healthy men . PNL suspended in nutrient medium did not practically ingest bacteria . The intake of bacteria got considerably intensified if leukocytes and bacteria ran into each other by turning over the test tubes during incubation . A still greater rise of phagocytic activity was discovered under the conditions favouring the attachment of PNL to the surface and the possibility of chemotaxis . These conditions were created by introducing a gelatin-coated film into the test tube.

Biochem J, 1983 Oct 1, 215(1), 183 - 9
Completion of the amino acid sequence of the alpha 1 chain from type I calf skin collagen . Amino acid sequence of alpha 1(I)B8; Glanville RW et al.; The complete amino acid sequence of the 279-residue CNBr peptide CB8 from the alpha 1 chain of type I calf skin collagen is presented . It was determined by sequencing overlapping fragments of CB8 produced by Staphylococcus aureus V8 proteinase, trypsin, Endoproteinase Arg-C and hydroxylamine . Tryptic cleavages were also made specific for lysine by blocking arginine residues with cyclohexane-1,2-dione . This completes the amino acid sequence analysis of the 1054-residues-long alpha (I) chain of calf skin collagen.

Am Rev Respir Dis, 1983 Oct, 128(4), 662 - 7
Effects of aging on antibacterial mechanisms in experimental pneumonia; Esposito AL et al.; Disease-free mature (6 to 8 months old) and senescent (24 to 26 months old) C57BL/6 mice were infected with Staphylococcus aureus and Klebsiella pneumoniae by peroral intratracheal inoculation . The lethal inoculum for 50% of animals (LD50) was identical for both groups of animals with Staphylococcus LD50 = 1.2 X 10(9) colony-forming units (cfu} and Klebsiella (LD50 = 2.3 X 10(8) cfu) . After challenges with high inocula of Staphylococcus (3 X 10(8) cfu) or Klebsiella (3 X 10(7) cfu), senescent mice more rapidly cleared viable organisms from the lungs than did the younger animals, and the differences were statistically significant at 48 h . The enhanced pulmonary clearance demonstrated by the older mice was associated with the recruitment of greater numbers of neutrophils into the bronchoalveolar spaces . When challenged with lower inoculums, mature mice cleared viable bacteria more rapidly than did the senescent animals, although a significant difference was observed only at 24 h after infection with 3 X 10(4) cfu Klebsiella . In contrast to high-inoculum infection in which greater than 90% of cells present in bronchoalveolar spaces were neutrophils, the predominant cells after low inoculum challenges remained alveolar macrophages in all groups . Differences in pulmonary clearance of viable bacteria in young and old mice were not associated with significant discrepancies in the rates of physical removal of 35S-methionine-labeled bacteria . Thus, the normal aging process results in alterations in the manner in which the host responds to pulmonary challenge with common bacterial pathogens, but these changes do not predispose to lethal infection.

Am J Clin Pathol, 1983 Oct, 80(4 Suppl), 603 - 8
Quality control of agar diffusion susceptibility tests: data from the Quality Assurance Service Microbiology program of the College of American Pathologists; Knowles RC et al.; Over a 12-month period, between July 1981 and June 1982, 115 active participants in the Microbiology program of the College of American Pathologists Quality Assurance Service (QAS) submitted a total of 555,619 individual determinations on three quality control reference strains using the NCCLS standardized disc diffusion procedure . Data is presented for those antimicrobic agent/reference strain combinations for which NCCLS control limits have been changed since the last report of QAS microbiology data or that continue to show discrepancies with current NCCLS individual daily test control guidelines . Data for Escherichia coli versus cefoxitin, doxycycline, and nalidixic acid and for Staphylococcus aureus versus cefoxitin, nafcillin, and oxacillin show good compliance with the new NCCLS guidelines and distributions that are all approximately Gaussian . Significant discrepancies were noted for six combinations; cefamandole, cephalothin, neomycin, and nitrofurantoin versus E . coli and amikacin and clindamycin versus S . aureus . Of these discrepancies, only neomycin/E . coli and amikacin/S . aureus can be accounted for by a subpopulation of laboratories, which, when removed, corrects the data.

Arch Dermatol, 1983 Oct, 119(10), 840 - 6
Staphylococcus aureus and atopic dermatitis; Dahl MV; Atopic dermatitis is a genetically determined skin disease that is strongly influenced by environmental factors . The skin of affected patients is usually colonized by large numbers of Staphylococcus aureus bacteria . These bacteria may aggravate atopic dermatitis or prevent resolution of the disease . The deleterious effects of S aureus on atopic dermatitis may be due to direct biologic action or may be due to indirect damage mediated by the immune and inflammatory systems.

J Clin Pathol, 1983 Oct, 36(10), 1120 - 8
Defective neutrophil function and microbicidal mechanisms in the myelodysplastic disorders; Martin S et al.; Neutrophil function studies have been carried out in a series of 44 patients with primary myelodysplastic syndromes (MDS) . In vitro tests of phagocytosis and killing of Candida guilliermondii and Staphylococcus aureus identified 13 patients with abnormal neutrophil function at presentation and a further 10 who developed abnormalities during the course of their disease . The incidence of defective function in the five disease categories in this series was: refractory cytopenia (RC) 8/17; refractory cytopenia with sideroblastic change (RC + SC) 5/8; acquired idiopathic sideroblastic anaemia (AISA) 2/4; refractory anaemia with excess blasts (RAEB) 7/11; chronic myelomonocytic leukaemia (CMML) 1/4 . Eleven of 23 patients with defective neutrophil function experienced severe infective complications; in only three of these patients were neutrophil counts less than 1 X 10(9)/l and susceptibility to infection was considered to reflect, at least partially, qualitative neutrophil abnormalities . There was no correlation between absolute neutrophil count and defective function . Abnormal overall neutrophil microbicidal activity was equally associated with impaired and normal phagocytosis . Some patients with intracellular killing defects had reduced myeloperoxidase (MPO) activities and one had reduced hexose monophosphate shunt (HMPS) activity . In two patients, whose neutrophils showed markedly impaired candidacidal activity, levamisole corrected function when added in vitro at 10(-7) M and also when administered in therapeutic dosage . It is suggested that deranged function, probably reflecting abnormalities in maturation of the granulocyte series, occurs across the myelodysplastic spectrum and that several microbicidal mechanisms may be defective.

Mikrobiyol Bul, 1983 Oct, 17(4), 267 - 9
{Bacteriophage typing of Staphylococcus aureus in Turkey}; Sevuk N et al.; S . aureus strains have been tested with several phages in Turkey . All of the phage groups of S . aureus strains were found in these investigations.

J Hyg (Lond), 1983 Oct, 91(2), 235 - 42
Enterotoxin production, phage typing and serotyping of Staphylococcus aureus strains isolated from clinical materials and food; Melconian AK et al.; The production of enterotoxins A, B, C and F by strains of Staphylococcus aureus isolated from various clinical sources and from isolates implicated in food poisoning was investigated . One hundred and ninety one of the 374 clinical strains (51.1%) were found to be enterotoxigenic; of these, 81 (27.7%) strains produced enterotoxin A, 57 (15.3%) strains produced enterotoxin B, 23 (6.2%) strains produced enterotoxin C, and 64 (17.1%) strains produced enterotoxin F . These enterotoxigenic strains were most frequently lysed by phages of group III (21.5%) or were not typable (22%) . Eighteen of the 29 strains implicated in food poisoning were enterotoxigenic . The correlation of antigens and bacteriophage patterns with enterotoxigenicity was determined: enterotoxin A being related to a4 antigen, enterotoxin B to phages of 94/96 complex with c1, o antigens, and enterotoxin F to phages of group I with 2632, k1k2, m antigens.

J Clin Immunol, 1983 Oct, 3(4), 392 - 8
Antigen-specific B-cell function in human autoimmune thyroiditis; De Bernardo E et al.; T-B cells from the peripheral circulation of patients with autoimmune thyroiditis were cocultured with sheep red blood cells (SRBC) or soluble human thyroglobulin (Tg), a self-antigen . The B-cell mitogen, Staphylococcus aureus, combined with macrophage-derived B-cell differentiating factor, induced in vitro lymphoid activation and proliferation in the presence or absence of Tg or SRBC, which was monitored after 6 days by specific anti-Tg and anti-SRBC plaque-forming cell (PFC) responses (expressed as PFC per 10(6) T-B cells) . In the presence of SRBC, significantly more anti-SRBC PFC (322 +/- 113, SE) were generated in normals (N = 5) compared with autoimmune thyroiditis patients (58 +/- 36) (N = 8) (P less than or equal to 0.001), data consistent with an antigen-specific T-cell defect . Anti-Tg PFC, not detectable in normal controls, were observed in patients in the absence (111 +/- 41) and presence (171 +/- 64) of Tg (10-1000 ng/ml) . However, variable responses were noted after such coculture experiments with Tg . Three patients demonstrated amplification of anti-Tg PFC, while three showed antigen-related inhibition of anti-Tg PFC . These data indicated heterogeneity of responses to Tg antigen in patients with autoimmune thyroid disease compounded by significantly depressed antigen-specific induction mechanisms.

J Immunogenet, 1983 Oct, 10(5), 379 - 93
Immunochemical evidence for multiple class I antigens coded by the MHC of the rat (RT1) and their differential expression on red blood cells and lymphocytes; Misra DN et al.; Five monoclonal antibodies reacting with class I MHC antigens were produced by fusing lymphocytes from WF (RT1u) rats immunized against DA (RT1a) rats with P3-X63-Ag8.653 myeloma cells . Sequential immunoprecipitation studies with the mAb and the WF anti-DA alloantiserum demonstrated the presence of four different class I molecules: all four molecules were reactive with the alloantiserum; three of them contained the determinant for mAb 155; two of the latter three molecules shared the determinants for mAb 3, 56 and 60, and one of these two molecules also contained the determinant for mAb 118 . The four molecules could be isolated from the antigen preparation by sequential immunodepletion first with 118, next with 3, then with 115 and finally with the alloantiserum or by sequential absorption with affinity columns of Sepharose 4B coupled to the antibodies . The three antigens which were sequentially isolated with the mAb 118, 3, and 155, respectively, were analysed by SDS-PAGE after digestion with Staphylococcus aureus V8 protease, and they showed differences in peptide patterns . The relative amounts of the antigens expressed on red blood cells and on lymphocytes were different based on the results of sequential isolation and indirect cellular radioimmunoassay: the antigen which reacted with both mAb 3 (and 56 or 60) and 155 was the major class I antigen on red blood cells, and the antigen which reacted with mAb 118, 3 (and 56 or 60) and 155 was the major class I antigen on lymphocytes.

Biochem J, 1983 Oct 1, 215(1), 147 - 51
Monoclonal antibody against the N-terminal end of human plasma fibronectin; Vartio T et al.; Purified human plasma fibronectin was digested with cathepsin G and the degradation products were tested for reactivity towards a monoclonal antibody . In an immunoblotting assay, after sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the digestion products, the 85 000-Mr and 72 000-Mr gelatin- and heparin-binding fragments as well as the N-terminal 30 000-Mr heparin-binding fragment reacted with the antibody, whereas the 64 000-Mr gelatin- and heparin-binding fragment did not . In enzyme immunoassay the antibody reacted with intact fibronectin and the 30 000-Mr fragment but not with a 40 000-Mr gelatin-binding fragment . The alignment of the binding domains in these fragments and in the intact molecule {Vartio (1982) Eur . J . Biochem . 123, 223-233} localizes the antigenic determinant to the 21 000 Da N-terminal Staphylococcus aureus-binding region of fibronectin.

J Biol Chem, 1983 Sep 25, 258(18), 11404 - 14
Purification and characterization of the 47,000-dalton protein phosphorylated during degranulation of human platelets; Imaoka T et al.; Secretion of platelet granule constituents is closely associated with the phosphorylation of a cytosol polypeptide of Mr = 47,000 that we have called P47 (Haslam, R . J., Lynham, J . A., and Fox, J . E . B . (1979) Biochem . J . 178, 397-406) . This polypeptide is a substrate of Ca2+-activated phospholipid-dependent protein kinase (Kawahara, Y., Takai, Y., Minakuchi, R., Sano, K., and Nishizuka, Y . (1980) Biochem . Biophys . Res . Commun . 97, 309-317) . Two-dimensional gel electrophoresis of protein from human platelets that had been preincubated with 32Pi demonstrated the presence under control conditions of 2-3 major forms of P47 that contained very little 32P (pI values, 6.6-6.8) and, after induction of secretion with thrombin, their replacement by 7-9 highly labeled phosphorylated forms of P47 (pI values, 6.1-6.5) . Native phosphorylated P47 was purified from thrombin-stimulated 32P-labeled platelets by ammonium sulfate fractionation and column chromatography on DEAE-cellulose, phenyl-Sepharose, and hydroxylapatite . The final 32P-labeled product was obtained in a yield of 20-25% and was purified about 400-fold relative to platelet lysate . This material was homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis but, like the starting material, contained 7-9 separable phosphorylated components with different pI values . Purified phosphorylated P47 had a sedimentation coefficient (s20,w) of 3.57 S and a Stokes radius of 3.33 nm from which an Mr = 49,000 and a frictional ratio (f/f0) of 1.4 were calculated . These findings and failure to detect multimers after treatment of the protein with dimethyl suberimidate indicate that P47 normally exists as a monomer . The 32P-labeled phosphate present in purified P47 had the chemical stability of serine or threonine phosphoesters and analysis indicated the presence of 83% phosphoserine and 17% phosphothreonine . Limited proteolysis of purified 32P-labeled P47 by Staphylococcus aureus V8 protease generated a major unlabeled fragment (Mr = 23,500) and up to six labeled fragments (Mr = 24,700-14,800), the relative amounts of the latter depending on the extent of proteolysis . The same labeled fragments were obtained after proteolysis of each of the major phosphorylated components of P47, suggesting that these represent different phosphorylation states of variants of the same protein and that most or all of the phosphorylation sites are on a single 14,800-Da segment of the protein . The availability of pure native phosphorylated P47 should facilitate investigation of the physiological role of this protein in platelets.

J Biol Chem, 1983 Sep 25, 258(18), 10924 - 33
Amino acid sequence of human beta-factor XIIa; Fujikawa K et al.; Human factor XII was activated by limited proteolysis with trypsin, and the resulting beta-factor XIIa (Mr = 30,000) was isolated by DEAE-Sephacel column chromatography . The complete amino acid sequence of beta-factor XIIa was then determined on peptides produced by enzymatic digestion with either trypsin, chymotrypsin, or Staphylococcus aureus V8 protease and by chemical cleavage at methionyl and tryptophyl bonds . beta-Factor XIIa is a glycoprotein composed of a heavy chain (243 amino acid residues) and a light chain (9 amino acid residues), and these two chains are held together by a disulfide bond . The carbohydrate is attached to asparagine residue 61 in the heavy chain . The amino acid sequence of the heavy chain shows a high degree of homology to the corresponding regions of other plasma serine proteases, such as plasmin, thrombin, factor IXa and factor Xa, as well as the pancreatic digestive enzymes . These results demonstrate that factor XII is the precursor of a typical serine protease that participates in the coagulation cascade.

J Biol Chem, 1983 Sep 25, 258(18), 11032 - 8
Purification from Escherichia coli of a periplasmic protein that is a potent inhibitor of pancreatic proteases; Chung CH et al.; A protein capable of inhibiting trypsin and other pancreatic proteases has been purified to homogeneity from Escherichia coli by conventional procedures and affinity chromatography . It is stable for at least 30 min at 100 degrees C and pH 1.0, but it is inactivated by digestion with pepsin . The inhibitor has an apparent molecular weight of 38,000 as determined by gel filtration and must be a homodimer since it contains a single 18,000-dalton subunit upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The inhibitor has an isoelectric point of 6.1 . One dimeric molecule of the inhibitor can bind two trypsin molecules to form a mixed tetrameric complex, in which trypsin molecules are completely inhibited . The inhibitor is not digested by the trypsin . When N-benzoyl-DL-arginine-p-nitroanilide was used as a trypsin substrate, half-maximal inhibition was observed at 22 nM . This protein also inhibits chymotrypsin, pancreatic elastase, rat mast cell chymase, and human serosal urokinase, but it does not inhibit human pulmonary tryptase, kallikrein, papain, pepsin, Staphylococcus aureus V8 protease, subtilisin, and thermolysin . Surprisingly, it did not inhibit any of the eight soluble endoproteases recently isolated from E . coli (i.e . proteases Do, Re, Mi, Fa, So, La, Ci, and Pi) nor the chymotrypsin-like (protease I) and trypsin-like (protease II) esterases in E . coli . The inhibitor is localized to the periplasmic space and its level did not change with different growth media or stages of cell growth . The physiological function of this E . coli trypsin inhibitor is unknown . We suggest that E . coli trypsin inhibitor be named "Ecotin."

Neurosci Lett, 1983 Sep 19, 40(1), 85 - 9
A hydrophobic dimer of acetylcholinesterase from Torpedo californica electric organ is solubilized by phosphatidylinositol-specific phospholipase C; Futerman AH et al.; A dimeric form of acetylcholinesterase from the electric organ of Torpedo californica was solubilized by phosphatidylinositol-specific phospholipase C from Staphylococcus aureus . The solubilized enzyme had a sedimentation coefficient of 7.3S which was not modified by detergents . The high salt-soluble asymmetric forms of acetylcholinesterase were not solubilized by the phospholipase . Our data suggest that the hydrophobic dimer of acetylcholinesterase may be associated with the plasma membrane through a specific interaction involving phosphatidylinositol.

J Biol Chem, 1983 Sep 10, 258(17), 10689 - 94
Mammalian beta-adrenergic receptors . Structural differences in beta 1 and beta 2 subtypes revealed by peptide maps; Stiles GL et al.; Photoaffinity labeling techniques using p-azido-m-{125I}iodobenzylcarazolol have recently demonstrated that both the beta 1- and beta 2-adrenergic receptor-binding subunits from mammalian tissues including heart, lung, and erythrocytes reside on peptides of Mr approximately equal to 62,000-64,000 . In this study, a two-dimensional gel electrophoresis method for peptide mapping was used to investigate and compare the structure of beta 1 - and beta 2-adrenergic receptor subtypes . When the photoaffinity labeled Mr approximately equal to 62,000 peptides from the beta 2-adrenergic receptors of rat lung and erythrocyte are subjected to simultaneous proteolysis using Staphylococcus aureus V8 proteinase or papain, exactly the same peptide fragments are generated from each subunit . In contrast, when the Mr approximately equal to 62,000 peptide containing the beta 1-adrenergic receptor-binding subunit derived from the rat heart is proteolyzed simultaneously with the Mr approximately equal to 62,000 peptide containing the beta 2-adrenergic receptors from either lung or erythrocyte, the peptide fragments generated are distinctly different . Peptide maps of beta 1-adrenergic receptors from the myocardial tissue of different species (pig versus rat) yield slightly different maps while the maps derived from the beta 2-adrenergic receptors of hamster lung and rat lung or erythrocytes reveal no interspecies differences . These data suggest: 1) alterations in the primary structure of the beta-adrenergic receptor may be responsible for the pharmacological specificities characteristic of beta 1- and beta 2-adrenergic receptor subtypes; and 2) alterations in the primary structure of similar beta-adrenergic receptor subtypes across different species may relate to the magnitude of their phylogenetic differences.

Arch Otolaryngol, 1983 Sep, 109(9), 624 - 6
Toxic shock syndrome related to nasal packing; Hull HF et al.; While toxic shock syndrome (TSS) is generally considered to be a tampon-related illness, an increasing number of reported cases have been related to surgical wounds . At least one case has been reported following nasal surgery with nasal packing . We report two additional cases of TSS associated with nasal packing . Because nasal packing is in some ways analogous to the use of tampons for menstrual hygiene, and nasal carriage of Staphylococcus aureus is frequent, the scarcity of TSS cases reported to occur following nasal packing is surprising . Otorhinolaryngologists are urged to report TSS cases associated with nasal packing to their state and local health departments or the Centers for Disease Control to aid in the understanding of the pathogenesis of this disease.

Yale J Biol Med, 1983 Sep-Dec, 56(5-6), 487 - 91
Clinical complications of Mycoplasma pneumoniae disease--other organs; Suzuyama Y et al.; Although self-limited respiratory tract infections caused by Mycoplasma pneumoniae are well recognized in children and young adults, respiratory involvements and hepatic dysfunction may occur . The frequency and clinical features of these complications were investigated . Experimental studies with regard to bacterial superinfection were also carried out . The test animals which were first infected with Mycoplasma pneumoniae and then with Staphylococcus aureus showed more extensive bacteriological and pathological changes than those infected with Staphylococcus aureus only . Liver biopsies performed on three human patients showed hepatic dysfunction and the histological findings were diagnosed as non-specific reactive hepatitis in each case.

Prikl Biokhim Mikrobiol, 1983 Sep-Oct, 19(5), 603 - 9
{Action of dioxidine on the membrane apparatus of the bacterial cell}; Rudzit EA et al.; A study of dioxydine effect on the membrane apparatus of Staphylococcus aureus 209 P and Micrococcus lysodeikticus revealed a decrease in the dehydrogenase activity of membrane preparations and in the specific concentration of cytochromes, if cells had been cultivated in a medium with subbacteriostatic concentrations of dioxydine . The total activity of the respiratory chain was slightly decreased . Conservatism of the membrane apparatus of the bacteria studied was demonstrated.

Antimicrob Agents Chemother, 1983 Sep, 24(3), 450 - 2
Comparative analysis of conjugative plasmids mediating gentamicin resistance in Staphylococcus aureus; Goering RV et al.; Five gentamicin-resistant clinical isolates of Staphylococcus aureus were found to contain self-transmissible plasmids of 32 to 37 megadaltons in size . Restriction endonuclease digests of the plasmids were markedly similar to those of reference plasmids of unrelated geographical origin, thus suggesting the significant contribution of common conjugal plasmids to the emergence of gentamicin resistance in S . aureus populations.

Antimicrob Agents Chemother, 1983 Sep, 24(3), 440 - 2
Effect of N, N'-dicyclohexylcarbodiimide and nigericin on Staphylococcus aureus susceptibility to gentamicin; Mandel LJ et al.; The abilities of the H+-ATPase inhibitor N, N'-dicyclohexylcarbodiimide and the antibiotic ionophore nigericin to enhance the bactericidal effect of subinhibitory concentrations of gentamicin in two strains of Staphylococcus aureus were studied . Each compound significantly increased both gentamicin uptake and killing . The use of agents which alter the plasma membrane energy state is a novel approach to enhance the activity of conventional antibiotics.

Plasmid, 1983 Sep, 10(2), 130 - 7
Staphylococcus aureus chromosomal mutation specifically affecting the copy number of Inc3 plasmids; Iordanescu S; A chromosomal mutation leading to an important increase in the copy number of plasmid pT181 and its derivatives has been isolated from Staphylococcus aureus strain 8325 . The amplification effect in the mutant strain SA1350 was found to be specific for plasmids of the Inc3 group, to which belongs pT181 . There are some other differences in the behavior of Inc3 plasmids between SA1350 and 8325, including stable maintenance in SA1350 at high copy number of temperature-sensitive replication mutants at restrictive temperatures, and altered incompatibility properties . Derivatives of SA1350 carrying only Inc3 plasmid mutants with high copy numbers (Cop mutants) could not be obtained, suggesting a lethal runaway plasmid replication in this situation . SA1350 expressed also a temperature-sensitive phenotype . The relationship of this character to the plaC1 mutation determining the amplification of Inc3 plasmids has not yet been elucidated.

J Reprod Med, 1983 Sep, 28(9), 611 - 4
Acute bacterial endocarditis in pregnancy . A report of three cases; Pastorek JG 2nd et al.; Acute bacterial endocarditis, a fulminating disorder most often caused by Staphylococcus aureus, is uncommon in pregnancy . However, the frequency of this disease may be increasing due to the prevalence of intravenous drug abuse . Three cases occurred during pregnancy at Charity Hospital, New Orleans; all three patients were intravenous drug abusers . One patient had polymicrobial disease.

J Clin Microbiol, 1983 Sep, 18(3), 683 - 8
Environmental study of a methicillin-resistant Staphylococcus aureus epidemic in a burn unit; Rutala WA et al.; During an outbreak of infections caused by methicillin-resistant (MR) Staphylococcus aureus in our burn unit, we conducted an extensive 10-week study to define the environmental epidemiology of the organism . The inanimate environment in patient rooms and adjacent areas was examined by using volumetric air samplers and Rodac plates . Airborne and surface level contamination with MR S . aureus was quantitated, and overall, MR S . aureus comprised 16, 31, and 40% of all bacterial growth from air, elevated surfaces, and floor surfaces, respectively . Mean air, elevated surface, and floor surface MR S . aureus contamination in rooms of MR S . aureus-infected burn patients were 1.9 MR S . aureus per ft3 (ca . 0.028 m3), 20 MR S . aureus per Rodac plate and 48 MR S . aureus per Rodac plate, respectively . Peak patient room environmental contamination levels were 6.9 MR S . aureus per ft3 of air, 70 MR S . aureus per Rodac plate per elevated surface and 138 MR S . aureus per Rodac plate per floor surface . Environmental contamination levels in the adjacent work areas were considerably lower than in infected patient rooms . There was ample opportunity for contamination of personnel through the inanimate environment in this unit.

J Virol, 1983 Sep, 47(3), 505 - 15
Nonstructural proteins of Semliki Forest virus: synthesis, processing, and stability in infected cells; Keranen S et al.; The synthesis of the nonstructural (ns) proteins of Semliki Forest virus was studied in vivo . The fourth ns protein, ns60, was identified and isolated . The order of translation (NH2-ns70-ns86-ns60-ns72-COOH) was determined by using various labeling procedures after or in the presence of a hypertonic block of translation initiation . A sequential labeling procedure was devised to specifically label defined segments of the polyprotein . The specific labeling procedures allowed isolation of the four ns proteins in radiochemically pure form by gradient polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate . The four ns proteins were shown to have different primary structures by digestion with V8 protease of Staphylococcus aureus . The processing of the ns polyprotein and the stability of the mature ns proteins were studied by pulse-chase experiments . The cleavage of each of the proteins from the polyprotein took place within 2 to 3 min after the translation of the polypeptide chain . The N-terminal protein, ns70, appeared in its mature form later than ns86, which follows it in the polyprotein, suggesting that ns70 undergoes a post-translational modification . The migration of the C-terminal protein, ns72, immediately after a pulse was slightly faster than after a chase, suggesting that ns72 also undergoes a post-translational modification other than a cleavage . The half-life of ns72 was shorter than that of the other ns proteins.

J Infect Dis, 1983 Sep, 148(3), 510 - 7
The contribution of human neutrophils and serum to host defense against Legionella micdadei; Weinbaum DL et al.; The interaction of Legionella micdadei with human polymorphonuclear neutrophils (PMNs) and serum was investigated to determine their roles in host defense against this organism . Serum and PMNs from normal donors having no antibody to L micadei were used . PMNs phagocytized once-agar-passaged (74.6% +/- 6.5%) and twice-agar-passaged (87.3% +/- 1.0%) L micdadei less (P less than 0.05) than L micdadei passaged multiple times on agar (97.5% +/- 1.0%) or Staphylococcus aureus (98.3% +/- 0.5%) . Under the same conditions, no phagocytosis of Legionella pneumophila occurred . Use of heat-inactivated serum abolished phagocytosis . PMN killing of once-agar-passaged (1.5% +/- 1.5%) and twice-agar-passaged (0) L micdadei was less (P less than 0.001) than that of L micdadei passaged multiple times on agar (88.6% +/- 4.0%) or S aureus (96.8% +/- 0.5%) . L micdadei was not killed by fresh serum, although, in contrast to L pneumophila, it was opsonized by C3 . Thus virulent L micdadei is phagocytized, but not killed, by human PMNs, with complement being the major opsonin.

South Med J, 1983 Sep, 76(9), 1131 - 5
Staphylococcus aureus bacteremia: a prospective study; Mylotte JM et al.; During a 22-month period, 47 patients with 49 consecutive episodes of Staphylococcus aureus bacteremia were identified and observed prospectively for the development of endocarditis and metastatic infection . Eighteen (37%) of the episodes were community-acquired and 31 (63%) were nosocomial . The mean patient age was 55 years, and all but nine patients had one or more underlying diseases . A primary focus was identified for 38 episodes (78%) most often an intravenous catheter, and 21 episodes (43%) were associated with a removable focus of infection . In this group, no patient had endocarditis after a mean duration of 20 days of therapy . Overall, two of 47 patients had endocarditis by clinical criteria; one was a drug abuser and one had no known heart disease . Forty-five of 49 episodes were treated with a single antimicrobial agent . There were 12 (24%) deaths in this series, seven (14.2%) directly due to staphylococcal infection . In this prospective study we found a low but definite risk of endocarditis associated with S aureus bacteremia . The mortality was similar to that in other recently published studies . All deaths occurred within two weeks of initiating therapy, indicating the potential importance of host factors in the outcome.

J Natl Cancer Inst, 1983 Sep, 71(3), 535 - 8
Prognostic indicators of tumor response to Staphylococcus aureus Cowan strain I plasma perfusion; Messerschmidt GL et al.; Ten tumor-bearing dogs were treated with passage of autologous plasma over fixed Staphylococcus aureus Cowan strain I . Five similar dogs were treated identically except for the exposure to S . aureus . These animals have been assessed to identify positive and negative prognostic variables for response . Nonresponder treated animals had significantly larger chest wall tumor bulk than did the responder and control groups (P less than .01) . Responder animals had fewer initial circulating immune complexes than did the nonresponders, though each group had similar reductions in immune complexes with therapy . Nonresponder animals had smaller volumes of plasma processed per kilogram of body weight per procedure than did controls (P = .016), whereas responder and control animals had similar volumes processed per kilogram of body weight per procedure (P = .84) . These data suggest that the response observed in our original series was significantly related to the larger amount of plasma treated per procedure and suggest that a factor may be eluted from the S . aureus cartridge that mediates this response.

Inflammation, 1983 Sep, 7(3), 269 - 76
Properties of neutrophils recruited into inflammatory foci with homologous or heterologous antigen in immunized ewes; Smith KL et al.; Studies were undertaken to determine functional properties of neutrophils attracted into involuted mammary glands of Staphylococcus aureus--immunized ewes by soluble antigens prepared from S . aureus (homologous antigen) or from Bacillus cereus (heterologous antigen) . The ewes were immunized with an S . aureus vaccine known to stimulate synthesis of cytophilic IgG2 antibody, and the inflammatory responses were elicited 4-8 weeks later by infusing homologous antigen into one gland and heterologous antigen into the other . Inflammatory cells were collected at 2, 4, and 8 h postinfusion . The magnitude of the cellular responses was similar in both glands with high proportions of viable neutrophils . There were no significant differences between neutrophil populations from each gland for proportions of cells bearing cytophilic immunoglobulin, although the proportions of cytophilic immunoglobulin-positive cells from both glands were lower in 2-h exudates than in 4-h or 8-h exudates . In invitro phagocytosis assays using 3H-labeled S . aureus and B . cereus no differences could be detected between the two populations of neutrophils in terms of phagocytic efficacy using a range of bacteria-neutrophil ratios and various opsonizing treatments.

Am J Dis Child, 1983 Sep, 137(9), 838 - 41
Neonatal infections . An important determinant of late NICU mortality in infants less than 1,000 g at birth; La Gamma EF et al.; For a 24-month period (1977 through 1978), the determinants of neonatal intensive care unit (NICU) mortality were examined retrospectively in 133 consecutively admitted newborn infants who weighed less than 1,000 g at birth . Seventy-one (53.4%) died during the first five days of life, 27 (20.3%) died after the first five days of life, and 35 (26.3%) were eventually discharged from the unit . Neonatal infection was the key determinant of increased mortality after the first five days of life . In these patients, gram-positive organisms were recovered from 54% (14/26) of all body fluid cultures from 21 infected infants . Infections included Staphylococcus epidermidis (27% {7/26}), Staphylococcus aureus (15% {4/26}), and Escherichia coli (11% {3/26}) . We propose that reducing the incidence of neonatal infections is necessary to substantially improve late NICU mortality in these very-low-birth-weight neonates.

Pathol Biol (Paris), 1983 Sep, 31(7), 593 - 5
{Value of the study of fibrinogen affinity in the identification of Staphylococcus aureus . Results of a multicenter study}; Carret G et al.; The passive haemagglutination reaction for detecting the Staphylococcus aureus fibrinogen affinity was done in a collaborative study . The test was compared to the major tests used in medical microbiology . The test was as efficient as the protein A or coagulase tests . Its simplicity, rapidity and specificity make it a useful tool for the diagnosis of this species.

Infect Immun, 1983 Sep, 41(3), 1105 - 11
Cloning, expression, and mapping of the Staphylococcus aureus alpha-hemolysin determinant in Escherichia coli K-12; Kehoe M et al.; A fragment of Staphylococcus aureus DNA encoding the alpha-hemolysin determinant was cloned from strain Wood 46 by inserting Sau3A-generated genomic DNA fragments between the BamHI sites of the lambda replacement vector L47.1 . Phages expressing alpha-hemolysin were detected by overlaying plaques formed from several thousand independent recombinant phage with erythrocytes and looking for zones of hemolysis . One phage expressing alpha-hemolysin was purified and named lambda w alpha 3 . This was subsequently shown to contain a 10.2-kilobase pair insert of S . aureus DNA . A 7.6-kilobase pair HindIII fragment encoding the alpha-hemolysin was subcloned from lambda w alpha 3 into the plasmid vector pACYC184 to form the hybrid plasmid pDU1148 . Escherichia coli K-12 cells harboring pDU1148 synthesized a low level of alpha-hemolysin which remained associated with the cells and was not secreted into culture supernatants . When the same strain was stabbed onto blood agar plates, no zones of hemolysis were detected after overnight growth at 37 degrees C but hemolysis developed if the plates were left at room temperature for 48 h . By introducing specific deletions or Tn5 insertions into plasmid pDU1148, the alpha-hemolysin gene was mapped to a region within a 3.3-kilobase pair EcoRI-HindIII fragment which was subcloned onto the vector plasmid pBR322 . A specific enzyme-linked immunosorbent assay with peroxidase-labeled rabbit anti-alpha-hemolysin antibodies was used to measure the levels of alpha-hemolysin antigen expressed in E . coli K-12 cells harboring pDU1148 or a variety of pDU1148::Tn5 and pDU1148 deletion mutants.

Infect Immun, 1983 Sep, 41(3), 1071 - 6
Impaired lung defenses against Staphylococcus aureus in mice with hereditary deficiency of the fifth component of complement; Cerquetti MC et al.; Mice of the C5-deficient DBA/2J and B10.D2/oSnJ inbred strains were aerosolized with Staphylococcus aureus, and the pulmonary clearance of bacteria was determined 4 h later . Both C5-deficient strains had a significantly decreased lung clearance of S . aureus compared with their genetically closest C5-sufficient relatives, DBA/1J and B10.D2/nSnJ strains, respectively . Serum hemolytic activity and pulmonary clearance of S . aureus were also investigated in F1 and F2 progenies (DBA/1J X DBA/2J and DBA/2J X B10.D2/oSnJ) . Serum hemolytic activity was present in all F1 (DBA/1J X DBA/2J) mice, and their pulmonary clearance of S . aureus was no different from that of the C5-sufficient parents (DBA/1J) . The absence of serum hemolytic activity (absence of C5) in all mice from F1 and F2 (DBA/2J X B10.D2/oSnJ) and 20% of the F2 (DBA/1J X DBA/2J) was related to a decreased lung clearance of S . aureus . These results are consistent with an autosomal recessive pattern of heredity for the murine abnormality in pulmonary clearance of S . aureus.

J Virol Methods, 1983 Sep, 7(3), 167 - 81
Demonstration of virus particles within immune complexes by electron microscopy; Lee FK et al.; A method has been developed to detect virus particles within immune complexes by electron microscopy (VICEM test) . Radioiodinated or unlabeled adenovirus type 2 immune complexes (IC) in human serum were used to determine the optimal conditions . Best results were obtained when the immune complexes were precipitated with 5% polyethylene glycol 6000, adsorbed with 0.5% of protein A-containing Staphylococcus aureus (Cowan 1) and eluted, with brief sonication, in 2 M propionic acid . When the eluate was examined by the pseudoreplica method of electron microscopy, morphologically intact viruses, concentrated over five-fold and free from background serum proteins, were readily demonstrated . The method is applicable to virus IC with a wide-range of antigen-to-antibody ratios and requires a minimum of 10(4)--10(5) virus particles . This method should be useful for the characterization of IC in clinical specimens obtained from patients with viral infections and for the possible demonstration of viral agents in immune complex diseases of unknown etiology . A modification of the technique can also be used for the rapid serological identification of viruses by electron microscopy, as exemplified by studies using adeno- and herpes simplex viruses.

Mol Cell Biol, 1983 Sep, 3(9), 1656 - 64
Reduced synthesis of pp60src and expression of the transformation-related phenotype in interferon-treated Rous sarcoma virus-transformed rat cells; Lin SL et al.; Treatment of Rous sarcoma virus-transformed rat cells with rat interferon-alpha (specific activity, 10(6) U/mg of protein) for 24 h caused a 50% reduction in intracellular pp60src-associated protein kinase activity . Staphylococcus aureus V8 protease digestion of pp60src, derived from 32P-labeled monolayer cultures incubated with or without interferon, revealed no differences either in the phosphopeptide pattern or in the phosphoserine-phosphotyrosine ratio . However, {3H}leucine pulse-labeling experiments showed that the synthesis of pp60src was reduced by 42 to 48%, relative to the level of bulk protein synthesis, in the interferon-treated cultures . Rat interferon-alpha also reduced the growth rate of Rous sarcoma virus-transformed rat cells in a dose-dependent manner over a 72-h period . The decrease in growth rate was accompanied by increases in the thickness and number of actin fibers per cell and by a decline in intracellular tyrosine phosphorylation by pp60src . The results suggest that interferon can inhibit the expression of the transformation-related phenotype by selectively reducing the synthesis of the Rous sarcoma virus transforming gene product . However, the interferon effects on the cytoskeletal organization and proliferation of Rous sarcoma virus-transformed cells may be due at least in part to the predominance of interferon-induced phenotypic changes over those caused by pp60src.

J Virol, 1983 Sep, 47(3), 600 - 10
Polyomavirus large T antigen binds independently to multiple, unique regions on the viral genome; Pomerantz BJ et al.; To map the polyomavirus large T antigen binding sites on the viral genome we employed a quantitative immunoassay . Defined, radiolabeled fragments of the viral genome were reacted with crude nuclear extracts prepared from lytically infected mouse 3T6 cells, and the fragments bound by large T antigen were immunoprecipitated with anti-T serum and Formalin-fixed Staphylococcus aureus . The immunoprecipitated DNA was then analyzed by gel electrophoresis and autoradiography . By the use of a variety of restriction endonuclease-generated fragments of wild-type and mutant viral DNAs, the region of high-affinity binding was localized to a 153-base-pair stretch between nucleotides 5292 and 152 . At least two independent binding sites lie within this region, one upstream and the other downstream of the Bg/I site at nucleotide 87 . One of the binding sites is located within sequences required in cis for DNA replication; the other overlaps the TATA box and cap sites of the early transcription unit . The two sites share a common sequence, A/TGAGGC-N4/5-A/TGAGGC, which may serve as the recognition sequence for large T antigen.

Am Rev Respir Dis, 1983 Sep, 128(3), 486 - 90
Decreased bactericidal function and impaired respiratory burst in lung macrophages after sustained in vitro hyperoxia; Suttorp N et al.; Lung macrophages (LM) play a crucial role in pulmonary bacterial defense . High inspired oxygen concentrations are used in a variety of diseases and "oxygen toxicity" could impair antibacterial function . We therefore examined the effect of sustained in vitro hyperoxia on LM bactericidal function, and on generation of two bactericidal oxygen metabolites . The LM were cultivated under aerobic (PO2 approximately 140 mmHg) or hyperoxic (PO2 approximately 630 mmHg) conditions for 48 h, and then incubated with Staphylococcus aureus labeled with 3H thymidine for 30 min . Incubated monolayers were processed for measurement of total bacterial uptake and for number of viable intracellular bacteria . Superoxide anion (O2-) and hydrogen peroxide (H2O2) generation was determined in similarly cultivated cells stimulated with opsonized zymosan . The results indicate that the bacterial killing capacity of oxygen-cultivated LM is significantly decreased (p less than 0.001) . In addition, a significant (p less than 0.001) decrease in generation of O2- and H2O2 was noted after exposure to high oxygen tensions . The data suggest that decreased bactericidal function after sustained hyperoxia may be due to an impairment of a specific bactericidal mechanism, i.e., an impaired "respiratory burst."

Endocrinology, 1983 Sep, 113(3), 1155 - 7
Structural similarities in the plasma membrane 3,3',5-triiodo-L-thyronine receptors from human, rat and mouse cultured cells . Analysis by affinity labeling; Cheng SY; Affinity labeling of Swiss 3T3-4 mouse fibroblasts with 0.3 nM N-bromoacetyl-3,{125I}3',5-triiodo-L-thyronine (BrAc{125I}T3) at 4 C for 0.5 h showed three labeled protein bands with apparent molecular masses of 55, 53 and 33 kilodaltons (kDal) in a ratio of 85:8:7 . Only the labeling of 55-kDal protein was selectively reduced to approximately 50% by 15 microM unlabeled T3 . Affinity labeling of the purified plasma membranes under identical conditions yielded similar results . One-dimensional peptide mapping by Staphylococcus aureus V8 or elastase digestion of the 55-kDal proteins from cells or plasma membranes gave identical peptide fragments . Thus the 55-kDal protein is a membrane-associated protein . Furthermore, affinity labeling of human epithelioid carcinoma A431 cells and purified plasma membranes gave similar results as those of Swiss 3T3 cells . Peptide mapping of elastase or S . auerus digestion of 55-kDal proteins from A431, Swiss 3T3 and GH3 rat pituitary tumor cells gave identical patterns . These results indicate that plasma membrane T3 receptors from three species have structural similarity in the hormone binding domains . Thus, the plasma membrane T3 receptor probably are highly conserved.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1983 Sep, 255(2-3), 234 - 8
Biotypes, serotypes and phage types of caprine strains of Staphylococcus aureus; Adegoke GO et al.; Nine of 11 caprine strains of Staphylococcus aureus examined belong to biotype C . They were typable serologically using antisera developed for typing human strains of Staphylococcus aureus . H2 was found to be the dominant thermostable antigen whilst 1 and 0 were the most common thermolabile antigens . Some strains possessed protein A and poly A beta (beta-glucosaminyl ribitol teichoic acid) . Phage typing of the strains was achieved with an international set of phages for typing human strains of Staphylococcus aureus and they belong to phage groups II, and the mixed group.

Clin Exp Immunol, 1983 Sep, 53(3), 663 - 71
In vitro immune cell function in six cases of immunoproliferative small intestinal disease after long term remission; Lopez-Botet M et al.; We have studied several parameters of in vitro immune cell function in peripheral blood mononuclear cells from six patients with immunoproliferative small intestinal disease after long term remission . We have observed two groups of patients with different patterns of response . (a) After stimulation with pokeweed mitogen (PWM) and Staphylococcus aureus, three patients showed a significant reduction of the Ig synthesis (A, G and M) and the proliferative response . In two of them, we found increased spontaneous suppressor T cell activity . In the third case, the diminished response could not be attributed, according to our assays, either to suppressor T cells, lack of T helper activity (although the number of OKT4+ cells was diminished) or an intrinsic B cell defect . The three patients showed normal or augmented NK activity and an inversion of the OKT4+/OKT8+ ratio was detected in two of them . (b) The remaining three patients showed a normal Ig synthesis after stimulation with PWM and a slightly depressed IgM synthesis in response to S . aureus . They expressed a normal T helper cell function and did not show increased spontaneous suppressor T cell activity . They had low levels of natural killer cytotoxicity and the OKT4+/OKT8+ ratio was not significantly altered . Taken together, our data indicate that significant alterations of the in vitro immune response can be found in peripheral blood mononuclear cells of some immunoproliferative small intestinal disease patients after long term remission.

J Hosp Infect, 1983 Sep, 4(3), 229 - 35
Total body bathing with 'Hibiscrub' (chlorhexidine) in surgical patients: a controlled trial; Leigh DA et al.; Total body bathing with 'Hibiscrub' (chlorhexidine-detergent) solution was compared with non-medicated soap in 224 patients admitted for surgery . Some 9.6 per cent of patients were found to be nasal carriers of Staphylococcus aureus on admission but 17.3 per cent were colonized at some time during their inpatient stay . Skin colonization by Staph, aureus was only seen in four patients (2 per cent), three were cleared by 'Hibiscrub' bathing but carriage persisted in the other patient who used non-medicated soap . A greater reduction in the total bacterial count on the skin and in the perianal region was seen in patients using 'Hibiscrub' . An increase in the bacterial count was frequently seen in patients using non-medicated soap . Postoperative staphylococcal wound infection occurred in nine patients (4-0 per cent) but nasal or skin carriage was only present in two patients . Although there was no difference in the rates of infection using 'Hibiscrub' or ordinary soap, pre-operative bathing with 'Hibiscrub' may be beneficial as there is a greater reduction in the total bacterial count . The use of non-medicated soap is of dubious value and may even increase the numbers of bacteria on the skin.

Clin Exp Immunol, 1983 Sep, 53(3), 703 - 8
Functional maturity of cord blood B lymphocytes: Staphylococcus aureus Cowan 1 induces IgG secretion in the human neonate; Unander AM et al.; Lymphocytes from newborn infants and adults were investigated with the polyclonal B cell activators Staphylococcus aureus Cowan 1, soluble protein A, and pokeweed mitogen (PWM) . In cord blood lymphocytes (CBL), protein A and PWM induced only meagre IgM responses, whereas Cowan 1 bacteria induced synthesis of both IgM and IgG in quantities comparable to adult levels . We were also able to establish the neonatal origin of the Ig producing cells . Thus, cord blood lymphocytes seem to be functionally mature enough to produce any Ig class, provided they are given the right stimulus.

Clin Exp Immunol, 1983 Sep, 53(3), 589 - 99
Detection of tumour associated antigen in eluates from protein A columns used for ex vivo immunoadsorption of plasma from melanoma patients by radioimmunoassay; Gupta RK et al.; Tumour associated antigen (TAA) of defined specificity and anti-TAA antibodies were isolated by elution with 0.1 M glycine-HCl buffer (pH 3.5) and 2.5 M MgCl2 from non-viable Staphylococcus aureus used in ex vivo immunoadsorption of plasma from four melanoma patients . The anti-TAA antibody activity in the MgCl2 eluate was determined by its ability to bind a melanoma 125I-TAA . The melanoma 125I-TAA was isolated and purified from the spent culture medium of a human melanoma cell line . The activity and specificity of TAA in the glycine-HCl eluates were determined by competitive inhibition in a radioimmunoassay in which melanoma 125I-TAA and an allogeneic antiserum obtained from a melanoma patient were used as the reagents . Results indicated that 0.04-0.81% of the total protein contained in the glycine-HCl eluates was TAA . The proportion of TAA to total protein in these eluates varied from patient to patient and treatment to treatment . Inhibition by the glycine-HCl eluates in the competitive radioimmunoassay was dose-dependent . Similarly, binding of melanoma 125I-TAA in a direct radioimmunoassay decreased with decreasing amounts of the anti-TAA antibody fraction . Quantitative analysis revealed that the MgCl2 eluates contained anti-TAA protein at levels ranging from 0.15 to 5.78% of total protein . Because both TAA and anti-TAA activities were found in eluates from S . aureus (protein A positive) used for immunoadsorption of plasma from melanoma patients, and because melanoma 125I-TAA isolated and purified from a human melanoma cell line did not bind to protein A directly, the results indicated that TAAs immunologically similar to the melanoma TAA were circulating in the form of immune complexes in plasma of four patients with melanoma and that these complexes could be removed from plasma by ex vivo immunoadsorption.

Postgrad Med, 1983 Sep, 74(3), 161 - 5, 168-72, 174-5
Breast cancer treatment--current status . 4 . Adjuvant therapy; Mitchell MS; Adjuvant chemotherapy of early breast cancer in premenopausal women has nearly reached the point where it can be said with confidence that the relapse-free survival rate, as well as overall survival rate, is being lengthened, especially when drug combinations are used . In postmenopausal women, however, adjuvant chemotherapy has not consistently improved the relapse-free rate . Some investigators have postulated that postmenopausal patients do not respond as well to chemotherapy as their premenopausal counterparts because they usually receive lower doses . Thus, trials of these drugs at doses that more closely approximate the calculated dose are needed in postmenopausal patients . Recent advances in determination of estrogen and progesterone receptors in breast cancer cells have allowed effective use of adjuvant hormone therapy instead of or in addition to chemotherapy in some patients with early disease . However, further investigation of the role of hormone manipulation in breast cancer is also needed . Adjuvant chemotherapy of advanced breast cancer, however, is only palliative; duration of relapse-free and overall survival is not nearly as long as in early disease . Hormone therapy has been effective as initial treatment in patients with positive estrogen and/or progesterone receptors . Alternative treatments under investigation include use of new anthracycline derivatives, high-dose single agents, and biomodulation with interferon and Staphylococcus aureus protein A . Thus, while advanced breast cancer is not now curable, the wide range of effective agents and promising new approaches to treatment certainly justify some optimism for the future.

J Exp Med, 1983 Sep 1, 158(3), 690 - 702
Selective suppression of an early step in human B cell activation by cyclosporin A; Muraguchi A et al.; The effect of cyclosporin A (CsA), a fungal metabolite with immunosuppressive properties, on the induction of human B cell proliferation and differentiation, has been described . CsA had a selective inhibitory effect on the activation phase of the cell cycle vs . the proliferation phase following preactivation of the cells . Cell enlargement and RNA synthesis of small resting B cells triggered by anti-mu were inhibited by addition of CsA (5-500 ng/ml) . The inhibitory effect of CsA was found only when the drug was added within 24 h of initiation of culture . In marked contrast, once small B cells were activated by anti-mu, the resulting large, activated B cells could be induced to initiate DNA synthesis by incubation with B cell growth factor (BCGF), and addition of CsA (1-1,000 ng/ml) to the culture did not suppress this BCGF-induced B cell proliferation . Addition of CsA to cultures of B cells which had been preactivated with Staphylococcus aureus Cowan strain I (SAC) and were already proliferating did not suppress B cell differentiation factor (BCDF)-induced differentiation of these cells . Thus, these data indicate that CsA can be used as a pharmacologic tool to dissect out human B cell responses into two distinct steps: (a) the initial activation step induced by anti-Ig, which is characterized by cell enlargement, RNA synthesis, and expression of receptors for BCGF; and (b) the proliferative step induced by BCGF in these preactivated B cells that undergo DNA synthesis and can then go on to differentiate in the presence of BCDF . In this regard, CsA selectively suppresses an early step of human B cell activation and has little inhibitory effect on the subsequent factor-dependent proliferation and differentiation.

J Chromatogr, 1983 Aug 26, 266, 157 - 62
Determination of immunoglobulins in blood serum by high-performance affinity chromatography; Crowley SC et al.; High-performance affinity chromatography columns were prepared by immobilizing protein A from Staphylococcus aureus on 10-microns diol-bonded silica . Immunoglobulin-containing samples were injected into the column at pH 7 and eluted by stepwise changes to pH 3 . Immunoglobulin G, A, and M standards were tested, but only a small fraction of the latter two was retained on the column . Analysis of 3-microliters samples of reference blood sera without pretreatment was performed using immunoglobulin G standards . Good agreement was observed between the immunoglobulin G concentrations measured by this method and by radial immunodiffusion . Chromatography of albumin and electrophoresis of retained serum fractions showed no interference by non-specific adsorption . Chromatographic analysis times were 4 min or less.

J Biol Chem, 1983 Aug 25, 258(16), 9973 - 81
Calcium-dependent proteolysis occurs during platelet aggregation; Fox JE et al.; Control and stimulated platelets were analyzed by two-dimensional polyacrylamide gel electrophoresis to determine whether proteins are altered during platelet activation . Platelets were stimulated with thrombin, collagen, or the calcium ionophore A23187, and aggregation was brought about by stirring in the presence of Ca2+ . These activated platelets contained at least three polypeptides not found in control platelets: 1) Mr = 200,000, pI between 6.2 and 6.4; 2) Mr = 100,000, pI = 6.3; and 3) Mr = 91,000, pI = 6.1 . An additional polypeptide, polypeptide 4, with Mr = 97,000 and pI = 5.9, was present only in platelets activated by thrombin . When aggregation was prevented, either by adding 5 mM ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) to the platelet suspension or by incubating the platelet suspension without stirring, polypeptides 1-3 were not formed . Partial hydrolysis of polypeptides 2 and 4 with Staphylococcus aureus V8 protease yielded distinct sets of peptide hydrolytic fragments . These differed from those produced by the hydrolysis of alpha-actinin, a major platelet protein, which has a molecular weight similar to polypeptides 2 and 4 . Polypeptides 1-3 were also produced during incubation of platelet lysates in the presence of Ca2+ . Generation of these polypeptides in lysates was prevented either by chelation of Ca2+ with EGTA or by the addition of N-ethylmaleimide, leupeptin, or mersalyl, inhibitors of the calcium-dependent protease . These data show that the calcium-dependent protease is activated during aggregation of platelets by physiological agents and suggest that this protease could have a role in platelet response to stimulation.

Br Med J (Clin Res Ed), 1983 Aug 20, 287(6391), 524 - 6
Systemic lupus erythematosus in Staphylococcus aureus hyperimmunoglobulinaemia E syndrome; Schopfer K et al.; The prevalence of autoimmune diseases, including systemic lupus erythematosus, is increased in failure of certain host defence mechanisms . Systemic lupus erythematosus, however, has not been recorded as a late complication of the Staphylococcus aureus hyperimmunoglobulinaemia E (hyper-IgE) syndrome . Such a case was investigated in a man suffering from a classic example of the syndrome . Antinuclear antibodies were analysed on a molecular basis . The emergence of immunological and clinical features of systemic lupus erythematosus in patients with defective host defence mechanisms against staphylococcal infections is unlikely to be fortuitous and may help elucidate the pathogenesis of systemic lupus erythematosus . The observations will also aid the long term management of patients with S aureus hyper-IgE syndrome.

J Biol Chem, 1983 Aug 10, 258(15), 9290 - 5
Structural properties of myosin from the particulate fraction of human blood platelets; Peleg I et al.; Fractionation of human blood platelets has revealed that myosin, a contractile and mechanochemical protein, is present in both the soluble and particulate fraction . The aim of this study was to elucidate whether platelets contain more than one myosin isoform, especially in view of the fact that in other cellular systems (cardiac muscle, amoeba) several myosin isoenzymes were found . The particulate fraction was solubilized by Triton X-100, and the myosin was purified by the same procedure used for the cytoplasmic myosin . The final preparation contained, in addition to myosin, a 130-kDa polypeptide, which was observed also in myosin preparations obtained from the soluble fraction . The electrophoretic mobilities of the two myosins were identical under both dissociating and nondissociating conditions . Comparison of the molecular structure of the heavy chain of the two myosins by limited proteolysis with Staphylococcus aureus V8 protease showed that the proteolytic fragments of the two myosins were rather similar, with only minor alterations in the quantitative distribution of the products . Two-dimensional peptide mapping of the iodinated tryptic peptides of the myosin heavy chains indicated that at least one peptide is missing in the map of the particulate myosin, as compared to its soluble counterpart . According to the two-dimensional peptide map, the 130-kDa polypeptide seems to be a proteolytic fragment of the myosin heavy chain and most probably the rod portion of the molecule . The observed minor variations in the structure of myosins isolated from the soluble and the fractions of human platelets may reflect differences in their respective physiological functions.

J Biol Chem, 1983 Aug 10, 258(15), 9043 - 5
Structure of linkage region between ribitol teichoic acid and peptidoglycan in cell walls of Staphylococcus aureus H; Kojima N et al.; The cell walls of Staphylococcus aureus H were found to contain mannosamine in an amount (28.1 nmol/mg) comparable to the content of muramic acid 6-phosphate . The acidic polymer fraction obtained by heating the cell walls at pH 2.5 was shown to contain mannosamine and glycerol in addition to the components of ribitol teichoic acid . Mild alkali treatment of this polymer fraction followed by gel filtration resulted in separation of a disaccharide N-acetylmannosaminyl(1 leads to 4)N-acetylglucosamine and the ribitol teichoic acid moiety that contained glycerol . Smith degradation of the reduction product from the same polymer fraction gave a fragment characterized as (1,2-ethylene-diol phosphate)-(glycerol phosphate)3-N-acetylmannosaminyl(1 leads to 4)N-acetylxylosaminitol . Thus, the ribitol teichoic acid chain in the cell walls is probably linked to peptidoglycan through a linkage unit, (glycerol phosphate)3-N-acetylmannosaminyl(1 leads to 4)N-acetylglucosamine.

J Biol Chem, 1983 Aug 10, 258(15), 9166 - 74
The complete amino acid sequence of rabbit muscle phosphoglucomutase; Ray WJ Jr et al.; The complete amino acid sequence of rabbit muscle phosphoglucomutase has been determined by isolating the 11 peptide fragments produced by the cyanogen bromide cleavage reaction and subjecting these to automated sequencing procedures . Products produced by treatment of some of these fragments with hydroxylamine, iodosobenzoic acid, mild acid, cyanogen bromide in formic and heptafluorobutyric acids, Staphylococcus aureus V8 protease, and trypsin (with or without blocking at lysine residues) were used to complete the sequence for each of the cyanogen bromide fragments . The cyanogen bromide fragments were ordered by isolating the four tryptic peptides produced by a limited tryptic digest of the native enzyme in the presence of its substrates and its bivalent metal ion activator, Mg2+, degrading these by means of trypsin, after blocking digestion at lysine residues, and isolating and identifying all fragments thus produced that contained 10 or more residues . The 561-residue sequence thus obtained is one of the longest that has been determined by chemical means . There is excellent agreement between this sequence and published compositions after appropriate normalization . The absorbance of the enzyme is about 7.0 at 278 nm for a 1% solution; this value is 9% lower than that previously used.

Eur J Immunol, 1983 Aug, 13(8), 605 - 14
Effect of protein A and its fragment B on the catabolic and Fc receptor sites of IgG; Dima S et al.; Radiolabeled protein A from Staphylococcus aureus (SpA) injected i.v . into mice and rabbits forms a soluble {(IgG)2-(SpA)1}2 complex (Mr = 684 000) which is identical in composition to that formed by SpA in vitro with an equivalent amount or an excess of IgG . A soluble rabbit IgG-SpA complex injected into a mice or rabbits dissociates completely in vivo and a new complex is formed with the IgG of the recipient animal . The half-life of SpA administered to a mouse or a rabbit is therefore the half-life of the IgG-SpA complex formed in vivo . In mice and rabbits the half-life of the complexes formed is 9 and 30 h, respectively, whereas the half-life of rabbit IgG in these animals is 106 and 153 h, respectively . Fragment B of SpA (fSpA) reacts with IgG of mouse and rabbit and forms an (IgG)1-(fSpA)1 complex . Complexes of identical composition are formed if fSpA is injected i.v . into mice and rabbits . The half-life of the complexes in mice and rabbits are much shorter than those of the corresponding free IgG in these animals (up to 15 times) . This result suggests that the binding of fSpA to the CH2 and the CH3 domains of IgG alters the function of the site, which controls the catabolism of IgG and is located in the CH2 domain . By contrast, fSpA does not change the Fc receptor-binding site of IgG, indicating that the Fc receptor site and the catabolic site are unrelated to each other.

Arch Ophthalmol, 1983 Aug, 101(8), 1275 - 7
Immune responses in rabbits with phlyctenules and catarrhal infiltrates; Mondino BJ et al.; Corneal phlyctenules and catarrhal infiltrates developed in rabbits immunized with Staphylococcus aureus cell walls (CWs) mixed with complete Freund's adjuvant (CFA) after topical challenge with viable S aureus . The immune responses of these rabbits to CWs and ribitol teichoic acid (RTA) before and after immunization with CW/CFA were studied and the results of skin tests with lymphocyte blastogenesis and hemagglutination tests correlated . After immunization of rabbits with CW/CFA, their lymphocytes showed a greater proliferative response to CWs in the lymphocyte blastogenesis test that correlated with delayed hypersensitivity reactions found in skin tests . Both tests suggested that the lymphocytes of immunized rabbits showed cellular immunity to CWs . Skin tests to RTA suggested an Arthus but not a delayed hypersensitivity reaction to this antigen . The results of skin tests and hemagglutination tests suggested that immunized rabbits expressed humoral and immunity to RTA, the major antigenic determinant of S aureus.

Ann Rheum Dis, 1983 Aug, 42(4), 362 - 7
Gas-liquid chromatographic analysis of synovial fluid: volatile short-chain fatty acids in septic arthritis; Borenstein DG et al.; Volatile short-chain fatty acids (SCFA) in synovial fluid from 80 patients were quantified by gas-liquid chromatography (GLC) . Characteristic patterns of volatile SCFA could not be associated with septic, nonseptic inflammatory, or noninflammatory groups . Mean concentrations of pentanoic and hexanoic acids were similar in all groups studied . In the septic arthritis group 3 of 4 patients with acetic acid and of 3 of 3 with 3-methyl butanoic acid had culture-proved staphylococcal infections . In patients with synovial fluid findings consistent with septic arthritis, including markedly raised leucocyte count, decreased glucose level, or detectable succinic acid, the detection of acetic and 3-methyl butanoic acids by GLC analysis may increase the suspicion that Staphylococcus aureus is the cause of the septic arthritis.

Am J Med, 1983 Aug, 75(2), 278 - 88
Cardiopulmonary toxicity in patients with breast carcinoma during plasma perfusion over immobilized protein A . Pathophysiology of reaction and attenuating methods; Young JB et al.; Tumoricidal reactions in dogs with spontaneous breast carcinoma occur after perfusion of plasma over protein A derived from Staphylococcus aureus and immobilized in collodion charcoal . When this treatment was extended to humans with breast cancer, hemodynamic and physiologic changes were noted . The evolution and spectrum of these reactions were evaluated during 47 plasma infusions in five patients . Initial treatment conditions consisting of rapid perfusion of plasma over high quantities of immobilized protein A were employed for 12 treatments in two patients . Within 30 minutes after treatments were begun, mean blood pressure, systemic vascular resistance, and stroke volume increased, as did heart rate, cardiac output, and rectal temperature; however, mean pulmonary artery pressure and total pulmonary resistance did not change . At 90 minutes, hypotension developed (lowest mean blood pressure was 59 +/- 14 mm Hg) that was associated with a decrease in systemic vascular resistance and total pulmonary resistance (536 +/- 66 and 146 +/- 44 dynes . second . cm-5, respectively) . Cardiac output increased, tachycardia developed, stroke volume decreased, and rectal temperature increased . During the hypotensive phase, values of creatinine clearance and fractional excretion of sodium diminished . Noncardiogenic pulmonary edema appeared occasionally, with bronchospasm noted once . No hemodynamic changes were seen when saline solution was passaged over protein A immobilized in collodion charcoal or when autologous plasma was given without passage over protein A immobilized in collodion charcoal . Treatment conditions were modified by diminishing protein A quantity and plasma volume and slowing plasma perfusion rate, which resulted in significant attenuation of all cardiopulmonary responses . This report, then, defines for the first time the physiologic basis of the cardiopulmonary toxicity in humans after plasma perfusion over immobilized protein A.

Obstet Gynecol, 1983 Aug, 62(2), 145 - 50
Risk factors in cesarean section infection; Hagglund L et al.; Factors associated with risk of postoperative infection after cesarean section were studied in 321 patients not given antibiotic prophylaxis . Infections occurred in 56 (25%) of the 228 patients who were delivered by emergency cesarean section and in eight (9%) of the 93 patients who underwent elective surgery (P less than .01) . These frequencies corresponded well with the infection rates reported after administration of antibiotics in other studies . Risk factors were: duration of operation more than one hour; blood loss more than 800 ml; presence of Staphylococcus aureus in the nares; signs of intrauterine infection before surgery; and failure of progress in labor . The results indicated that obstetric interventions had been performed more frequently in patients at risk of infection, rather than being the real cause of the infections . The importance of strict preoperative hygienic routine is discussed.

Zh Mikrobiol Epidemiol Immunobiol, 1983 Aug, (8), 36 - 8
{Antagonism between hospital strains of Staphylococcus aureus and lactic acid bacteria in vitro and the use of the latter as a sanitary agent}; Ambartsumian AD et al.; The study of antagonism between S . aureus hospital strains and lactic acid bacteria, strain 317/402 "Narine", revealed that the latter possessed high antagonistic activity . A new method for the sanitation of carriers of S . aureus hospital strains was developed; this method made it possible to limit the epidemiological significance of 82% of these strains.

J Ethnopharmacol, 1983 Aug, 8(2), 149 - 61
The antibacterial properties of an Aztec wound remedy; Davidson JR et al.; Wound treatment practices of the Aztecs are discussed . The use of concentrated maguey sap (Agave ssp.) was widely dispersed and has persisted in folk medicine . A possible reason may be that it is effective . Laboratory analysis of maguey syrup indicates that its utilization as a remedy by ancient and modern Mesoamericans could contribute to the healing process of aerobic wound infections . Both pyogenic and enteric bacteria appear to be susceptible to maguey syrup . The traditional addition of salt to the remedy seems to enhance the effectiveness of the material in inhibiting the growth of one of the major causes of pus-forming or pyogenic infective processes, Staphylococcus aureus . This finding is additional proof of the effectiveness of pre-Hispanic medicine, and of the skills of pre-Hispanic physicians.

Vet Microbiol, 1983 Aug, 8(4), 397 - 404
Encapsulation of Staphylococcus aureus isolated from bovine milk; Norcross NL et al.; Evidence is presented that nearly all Staphylococcus aureus infections of the bovine mammary gland are by encapsulated organisms (94% of isolates examined) . This observation is based on the demonstration of diffuse growth in serum-soft agar, of cultures taken directly from mastitic milk without subculture on artificial media . Only milks containing pure cultures of S . aureus were examined . It was previously reported that only 6% had capsules . Evidence is presented that as few as 3 or 4 subcultures and/or a short storage on artificial media results in loss of encapsulation of most S . aureus of bovine origin . Antisera raised against encapsulate strains inhibit the expression of capsule formation on bacteria that are encapsulated in the presence of normal serum.

J Gen Microbiol, 1983 Aug, 129 (Pt 8), 2421 - 8
Survey of taurine uptake and metabolism in Staphylococcus aureus; Smiley DW et al.; Taurine has been reported to be a component of the capsular polysaccharide of the encapsulated M strain of Staphylococcus aureus . This led to a study of the uptake and metabolism of {1,2-14C}taurine in a variety of encapsulated and unencapsulated S . aureus strains . Taurine was taken up by all strains studied . A discrepancy between uptake measured as depletion of radioactivity from growth medium and as cell-associated radioactivity suggested that taurine may be catabolized to CO2 in some strains . In most strains, cell-associated radioactivity was located mainly in cold TCA-soluble (pool metabolites) fractions . About 90% of the cell-associated radioactivity was present in the pool metabolites fraction in the M strain, and about 10% in hot TCA-soluble (nucleic acid-teichoic acid-capsular polysaccharide) fraction . Radioactivity in spent medium and the capsular polysaccharide-containing fraction appeared to be present as taurine in this strain . Radioactivity in the pool metabolites fraction of three of the strains examined did not chromatograph as taurine, indicating that taurine was converted into other cell metabolites . One strain incorporated radioactivity from taurine into cellular macromolecules, thus revealing a heterogeneity of staphylococcal taurine metabolism.

Can J Ophthalmol, 1983 Aug, 18(5), 213 - 6
Management of acquired dacryocystitis; Hurwitz JJ et al.; Acquired dacryocystitis that has its onset after the first year of life is due to fibrotic obstruction within the lacrimal drainage pathways that is usually secondary to trauma or infection . Staphylococcus aureus is the most common pathogen, but attempts should be made to express or aspirate material from the lacrimal sac for culture and antibiotic sensitivity testing . When the strain is resistant to penicillin or when a culture specimen is not obtainable, cloxacillin is the antibiotic of choice . The timing of application of hot compresses is important so that the swelling can be localized to the sac but perforation will not be induced . Once the inflammation has resolved, dacryocystorhinostomy may be needed to drain residual fluid . Dacryocystography is useful to indicate whether there is persistent obstruction or stenosis; if the results are normal but symptoms persist, scintiscanning may demonstrate a delay of tear flow out of the sac or may reveal a stone.

Can J Biochem Cell Biol, 1983 Aug, 61(8), 892 - 905
The triterpenoid carotenoids and related terpenoids in Staphylococcus aureus 209P; Taylor RF et al.; The carotenoids of Staphylococcus aureus 209P have been identified as triterpenoid (C30) carotenoids . The pigments include members of a carotene dehydrogenation sequence with C30 analogs of phytoene, phytofluene, zeta-carotene, and neurosporene . The major xanthophyll present under the cultural conditions used is a carotenoic acid derived from the C30 analog of neurosporene . A cis-isomer of this major xanthophyll has also been characterized, as have four other xanthophylls . Of the latter xanthophylls, two have been identified as the hydroxy and glycosyloxy derivatives of the major xanthophyll, while the remaining two xanthophylls are carotenoid esters . Squalene and menaquinone MK-8 have also been identified in the bacterium.

Appl Environ Microbiol, 1983 Aug, 46(2), 515 - 7
Prevention of Staphylococcus aureus lysis; Bhaduri S; Staphylococcus aureus S-6 cells grown in chemically defined media often lysed after exponential growth . Lysis could be prevented by the addition of alanine or proline before the culture reached stationary phase.

Am J Vet Res, 1983 Aug, 44(8), 1433 - 41
Cytologic observations of the bovine teat end; Nickerson SC et al.; Cells infiltrating from the vasculature and histologic components of internal tissues of teats (mammary papilla) from noninfected udder quarters were studied, using light and electron microscopy . Morphometric analysis demonstrated a progressive increase in number of infiltrating cells from the distal teat cistern (sinus papillaris) to the junction of the Furstenberg's rosette (distal termination and convergence of mucosal folds lining the teat cistern) and the streak canal (ductus papillaris) . Plasma cells contributed to cellular increases in subepithelial connective tissue and were the most prevalent infiltrating cell type . Plasma cells also penetrated the basal epithelial lining of the rosette area and occasionally migrated to the luminal surface near the squamocolumnar junction . Neutrophils and monocytes contributed to the increase in cells infiltrating the epithelial lining . Few infiltrating cells were observed in epithelium and underlying stroma of the streak canal . Cytologic comparison demonstrated a reduction in all cell types from lactating to involuting phases of lactation . Greater numbers of plasma cells, lymphocytes, and monocytes were observed in teat end tissues from quarters previously infected with Staphylococcus aureus.

Acta Pathol Microbiol Immunol Scand {B}, 1983 Aug, 91(4), 273 - 8
Investigation of Micrococcaceae in a department of cardiac surgery . Biochemical characterization and sensitivity patterns of strains isolated from patients, staff, and air; Hansen BG; A total of 965 strains of Micrococcaceae isolated from 200 patients, personnel, and air in a department of cardiac surgery were classified by means of Baird-Parker's scheme . The majority of strains were identified as Staphylococcus epidermidis (S . epidermidis) biotype 1, but S . epidermidis biotype 4 accounted for c . 25% of isolates from patients post-operatively . Pre-operative isolates were generally sensitive to most antibiotics tested while post-operative strains of coagulase-negative Micrococcaceae from patients and isolates from personnel and air were frequently multiply-resistant . Strains of Staphylococcus aureus (S . aureus) were sensitive or resistant only to penicillin . More patients were colonized with coagulase-negative Micrococcaceae after operation than at admission to the hospital (p less than 0.001), while the frequency of S . aureus carriers was the same before and after operation and equal to the frequency found earlier . The frequency of S . aureus carriers among the personnel, however, was lower than reported earlier (10%) . Multiply-resistant strains of S . epidermidis seem to have replaced resistant strains of S . aureus as the predominant hospital saprophyte among Micrococcaceae.

J Clin Microbiol, 1983 Aug, 18(2), 408 - 15
In vitro response of Staphylococcus aureus from cystic fibrosis patients to combinations of linoleic and oleic acids added to nutrient medium; Campbell IM et al.; The effect of supplementing nutrient substrate with various combinations of concentrations of oleic and linoleic acids on the growth of 11 strains of Staphylococcus aureus was assessed . Whereas increasing the concentration of linoleic acid by itself greatly diminished the growth of all 11 strains, concomitant increases in oleic acid greatly diminished the inhibitory effect of linoleic acid . With oleic acid in the nutrient substrate, most of the strains were induced to produce slime which surrounded the cells . Since the slime incorporated oleic but not linoleic acid, such slime production isolated the cells from direct contact with the growth inhibitor, linoleic acid.

J Appl Bacteriol, 1983 Aug, 55(1), 81 - 8
The protective effect of some food ingredients on Staphylococcus aureus MF31; Hurst A et al.; The upper limiting temperature of growth of Staphylococcus aureus MF31 in heart infusion broth (HI) was about 44 degrees C but addition of monosodium glutamate (MSG) and soy sauce permitted the organism to grow above this temperature . This effect is similar to that of NaCl . Tomato ketchup, Worcestershire and HP sauces added to HI did not allow growth at the non-permissive temperature of 46 degrees C but death was delayed . Staphylococcus aureus died in unsupplemented chicken meat slurry at 46 degrees C but grew at 48 degrees C in slurry supplemented with 5.8% NaCl and survived incubation for 18 h at 50 degrees C in slurry supplemented with 5.8% NaCl and 5% MSG . Cultures grown at 37 degrees C had a D60 value of 2 min in 50 mmol/l Tris (pH 7.2) buffer . Cultures grown at 46 degrees C in HI containing 5.8% NaCl had a D60 value of 8 min in Tris buffer . Addition of 5.8% NaCl plus 5% MSG to the buffer increased the D60 by a factor of about 7 for both cultures . In storage experiments at room temperature, the culture grown at 37 degrees C and at 46 degrees C plus 5.8% NaCl died at about the same rate in salami . In milk powder, however, the count of 37 degrees C culture decreased from 10% g to 10(6)/g in 5 weeks while the count of 46 degrees C culture remained unchanged . In cottage cheese, freeze-dried rice and macaroni, the 37 degrees C cultures also died more rapidly . It is suggested that cultures grown at 46 degrees C plus 5.8% NaCl may be suitable for experiments with artificially contaminated foods.

Clin Nephrol, 1983 Aug, 20(2), 85 - 8
Vancomycin serum levels and toxicity in chronic hemodialysis patients with Staphylococcus aureus bacteremia; Masur H et al.; The pharmacokinetics and toxicity of six week courses of vancomycin were assessed prospectively in 12 chronic hemodialysis patients who had 17 episodes of Staphylococcus aureus bacteremia . Patients were treated with 1 gram doses of vancomycin at weekly intervals for six weeks . Peak serum vancomycin concentrations ranged from 5.5-40.0 micrograms/ml and trough concentrations were 1.0-12.0 micrograms/ml at the time of the second dose . No patients demonstrated important drug accumulation at the time of the fifth dose . Pure tone audiometry demonstrated no auditory toxicity . Flushing and pruritus (two patients) were only adverse effects noted . In 16 episodes blood cultures were sterilized within 48 hours of therapy . This investigation demonstrates that in chronic hemodialysis patients with S . aureus bacteremia vancomycin is a safe and microbiologically effective antimicrobial agent . Peak and trough serum concentrations vary widely when 1 gram doses are given at one week intervals, and thus it is recommended that concentrations be measured for each patient, particularly if the minimum bactericidal concentration of vancomycin for the clinical isolate is greater than 1.0 microgram/ml.

Acta Ophthalmol (Copenh), 1983 Aug, 61(4), 567 - 75
Management of complications after cataract surgery by the pars plana approach; Pedersen OO; Various ocular complications following cataract surgery were treated using vitrectomy instruments by the pars plana approach . A total of 11 eyes were operated . Eight of these eyes had various anterior segment problems: Secondary cataract (2 eyes), cyclitic pupillary membranes (2 eyes), up-drawn pupils (3 eyes), and occluding pupillary membrane following haemorrhage (1 eye) . In 2 of these eyes opacities in the posterior vitreous were present as well . In addition, pars plana vitrectomy was performed in 3 eyes with presumed bacterial endophthalmitis . In one of these eyes, the vitreous was sterile at the time of the vitrectomy . The vitreous of the 2 other eyes contained Staphylococcus epidermidis, and Staphylococcus aureus, respectively . In the eyes with anterior segment problems, clear optical media were obtained in all cases . The visual outcome in these patients was determined by the condition of the retina at the time of surgery . Two of the eyes with endophthalmitis were saved with some preservation of vision . The third of these eyes which suffered from endopthalmitis caused by penicillinase-producing Staphylococcus aureus, was lost . In this case vitrectomy and adequate antibiotic therapy was delayed . The modern vitrectomy instruments designed for pars plana vitrectomies are highly suitable for management of anterior segment problems following cataract surgery . When these problems are combined with pathology of the posterior vitreous, both kinds of problems may be dealt with at the same operation . In cases of presumed bacterial endophthalmitis, early vitrectomy should be considered to remove infective organisms, to obtain a definite diagnosis, and at the same time the eye may be perfused with antibiotics.

J Gen Microbiol, 1983 Aug, 129 (Pt 8), 2457 - 65
The effect of membrane-bound beta-lactamase on linoleic acid sensitivity in Staphylococcus aureus; Greenway DL et al.; The presence of a plasmid conferring resistance to penicillin (PC plasmid, e.g . pI258blaI-) in Staphylococcus aureus NCTC 8325 increases the sensitivity of such a bacterium to the growth inhibitory effects of linoleic acid, whereas a plasmid conferring resistance to tetracycline does not affect linoleic acid sensitivity . The increased linoleic acid sensitivity of bacteria containing a PC plasmid may be related to the penicillinase protein itself since (i) strains having inducible penicillinase show increased sensitivity only after induction, (ii) strains in which penicillinase is directed from chromosomal or plasmid-borne genes show similar increased linoleic acid sensitivity and (iii) notwithstanding the above, the linoleic acid inhibitory effect is enhanced in a strain in which penicillinase activity is greatly reduced by a point mutation in the structural gene for penicillinase . The enhanced linoleic acid sensitivity seems to require the membrane-bound penicillinase since added extracellular penicillinase does not confer this sensitivity, and there appears to be a specific interaction between the membrane-bound penicillinase activity and linoleic acid.

Br Heart J, 1983 Aug, 50(2), 193 - 5
Dehiscence of infected aortocoronary vein graft suture lines . Cause of late pseudoaneurysm of ascending aorta; Smith P et al.; Two cases are described of late pseudoaneurysm of the ascending aorta caused by dehiscence of infected aortocoronary vein graft suture lines . Both cases were associated with early postoperative Staphylococcus aureus superficial wound infection . This complication should be suspected in patients developing protracted wound infections after operation, particularly when associated with rigors.

J Natl Cancer Inst, 1983 Aug, 71(2), 325 - 30
Cytotoxic responses induced by peptidoglycans with or without in vivo antitumor activity; Goguel AF et al.; Cytotoxic responses mediated by effector cells stimulated in vivo were studied after ip injection in mice of peptidoglycans extracted from gram-positive bacteria . A comparison was done between Staphylococcus aureus peptidoglycan (PGS), which possessed an antitumor effect, and Micrococcus lysodeikticus peptidoglycan, which was ineffective against tumors . Both peptidoglycans induced similar effects on the modulation of T-cell cytotoxic response . Both were able to stimulate splenic and peritoneal nonspecific cytotoxicity against YAC-1 lymphoid tumor cells, but only PGS could induce cytotoxicity and cytostasis against solid tumor target cells.

J Immunol, 1983 Aug, 131(2), 801 - 5
The effects of interleukin 1 on human B cell activation and proliferation; Falkoff RJ et al.; The precise role of B cell surface immunoglobulin (slg) in the activation of B cells is unclear at present . In particular, it is uncertain whether ligands interacting with the B cell slg suffice to induce proliferation, or simply induce a state of activation in which the B cell becomes responsive to growth factors made by accessory cells . We have examined the effects of two ligands, Staphylococcus aureus Cowan strain I (SAC) and antihuman mu chain (anti-mu), which interact with B cell slg on highly purified human peripheral blood and tonsillar B cells cultured at low cell concentrations . The effects on B cell proliferation of these ligands alone or in combination with highly purified interleukin 1 (IL 1) or a supernatant of a human T-T hybridoma containing a B cell growth factor (BCGF) were studied . SAC with its high cell wall content of protein A triggered maximal B cell proliferation which was not increased further by IL 1 or BCGF . High concentrations of soluble F(ab')2 fragments of goat anti-mu chain also induced significant B cell proliferation . Lower concentrations of anti-mu resulted in little or no B cell proliferation but activated the B cell to a state of responsiveness to both IL 1 and BCGF . IL 1 by itself had no effect on the proliferation of unstimulated B cells or on the proliferation of in vivo-activated B cells which responded to BCGF in vitro, but demonstrated clear synergy with low concentrations of anti-mu antibody . BCGF alone augmented the proliferation of unstimulated B cells, presumably by acting on B cells which had undergone some degree of activation in vivo . In addition, it showed marked synergy with anti-mu antibody, which resulted in proliferation similar in magnitude to that induced by SAC . This synergy was far greater than that seen between anti-mu antibody and IL 1, and the resulting proliferative response was only slightly increased by the presence of IL 1 . We conclude that the importance of accessory cell factors for the initial rounds of B cell proliferation depends on the strength of the initial slg-mediated activation signal . When this is strong, the response is maximal and independent of accessory cells or accessory cell factors . When it is suboptimal, a moderate synergy is seen with IL 1 and a dramatic synergy with BCGF.

Acta Pathol Microbiol Immunol Scand {B}, 1983 Aug, 91(4), 279 - 84
Population analysis of susceptibility to methicillin, vancomycin, and three cephalosporines in two methicillin-resistant strains of Staphylococcus epidermidis; Hansen BG; Population analyses of susceptibility to methicillin, vancomycin, cephalothin, cefamandole, and cefoxitin were carried out with two strains of Staphylococcus epidermidis (S . epidermidis) resistant to penicillin, streptomycin, and methicillin . Two strains of Staphylococcus aureus (S . aureus), one sensitive to penicillin the other resistant, were used as controls . All strains were clinical isolates . From the strains of S . epidermidis it was possible to select sub-populations highly resistant to methicillin, cephalothin, cefamandole, and cefoxitin . The mutation-rate varied between c . 10(-3) and 10(-7) . The two strains were homogeneous as regards vancomycin susceptibility . Methicillin was shown to be more penicillinase-stable than the cephalosporines . Treatment in vivo, with cephalothin and vancomycin did not increase the frequency of highly resistant bacteria against any of the antibiotics, but during treatment with cephalothin the majority-population became eight times more resistant to vancomycin and may explain why the treatment in vivo was not successful.

J Antibiot (Tokyo), 1983 Aug, 36(8), 1045 - 50
Immunoactive peptides, FK-156 and FK-565 . I . Enhancement of host resistance to microbial infection in mice; Mine Y et al.; The protective effect of an immunoactive peptide, D-lactoyl-L-alanyl-gamma-D-glutamyl-(L)-meso-diaminopimelyl-(L)-glycine (FK-156) and a related compound, heptanoyl-gamma-D-glutamyl-(L)-meso-diaminopimelyl-(D)-alanine (FK-565) was determined in mice with various kinds of microbial infections . FK-156 and FK-565 were given to mice either subcutaneously or orally before challenge . The drugs enhanced significantly the defense of mice against acute systemic infections induced by various extracellular and facultative intracellular organisms, and subcutaneous abscess by Staphylococcus aureus . The protective effect of these drugs against Escherichia coli infection differed considerably depending on the route of administration; FK-156 was only effective by the parenteral route; however, FK-565 was effective by both parenteral and oral routes . After subcutaneous dosing with FK-156, the enhancement of host defense of mice against E . coli infection was more rapid than against Listeria infection . The enhancing effects of FK-156 and FK-565 on host defense of mice against pseudomonal infection was more potent than other immunoactive drugs.

Virology, 1983 Aug, 129(1), 212 - 7
An immunological focus assay for murine leukemia viruses on viable attached cell lines; Evans LH et al.; An assay is described for the detection and isolation of murine leukemia virus (MuLV)-infected cells in viable monolayers . The procedure utilizes antisera or monoclonal antibodies which specifically bind cell surface viral antigens of infected cells . Bound antibodies are subsequently detected by binding with 125I-labeled Staphylococcus aureus protein A followed by autoradiography of the tissue culture vessel . Focal areas of infection can be identified from the autoradiograph and infected cells can subsequently be isolated and subcultured as MuLV-producing cell lines . With appropriate antibodies the procedure should be useful for the direct isolation of minor components, including mutants, in complex virus mixtures.

Virology, 1983 Aug, 129(1), 127 - 36
Immunoprecipitating human antigens associated with vesicular stomatitis virus grown in HeLa cells; Little LM et al.; Vesicular stomatitis virus (VSV) preparations made in HeLa cells, VSV(HeLa), appeared to contain non-viral structural proteins . This was suggested by neutralization of the virus with homologous and heterologous antisera made against VSV prepared in different cells . Antisera against uninfected HeLa cells failed to neutralize VSV(HeLa) but did immunoprecipitate the virus in the presence of Staphylococcus aureus . These immunoprecipitated VSV(HeLa) retained their infectivity, despite the presence of antibody and bacteria . The anti-HeLa cell serum did not react with VSV grown in rodent cells nor did anti-Vero cells serum immunoprecipitate VSV(HeLa) . When the anti-HeLa cell serum was absorbed with whole HeLa cells, it no longer specifically precipitated VSV(HeLa) . Because over 98% of infectious VSV(HeLa) was neutralizable by anti-VSV serum and immunoprecipitable by anti-HeLa serum, these virions were called mosaics . Physical identification of HeLa cell determinants on the mosaics was accomplished by further purification and radioiodination followed by selective immunoprecipitations with antisera . Two to three major bands with molecular weights around 75,000 Da were identified as HeLa cell determinants associated with the mosaic VSV(HeLa).

Pediatr Res, 1983 Aug, 17(8), 657 - 61
Pulmonary clearance of Staphylococcus aureus and plasma angiotensin-converting enzyme activity in hydrocarbon pneumonitis; Nouri LA et al.; Pulmonary clearance of Staphylococcus aureus and plasma angiotensin-converting enzyme (ACE) activity were investigated in an experimental mouse model of kerosene aspiration . Twenty-four hours after acute kerosene aspiration, mice were exposed for 30 min to an aerosol containing the pathogen, and the uncleared bacteria ratio (UBR) determined 4 h after nebulization . The results showed a significant increase (P = 0.004) in UBR in animals with severe pneumonitis (0.44 +/- 0.05) when compared with controls (0.24 +/- 0.03) . This impairment in lung clearance correlated with the increase in lung weight and the decrease in plasma ACE levels . Mice with kerosene pneumonitis had zones of lung injury, and areas with no gross signs of tissue damage . Lung clearance of S . aureus was significantly impaired in damaged areas whereas it was no different from controls in the non-affected areas . It is suggested that the measurement of plasma ACE activity may be an adjunct in the assessment of the extent of lung injury in hydrocarbon aspiration in children.

J Cell Biol, 1983 Aug, 97(2), 368 - 77
Functional activity of enucleated human polymorphonuclear leukocytes; Roos D et al.; Enucleated human polymorphonuclear leukocytes (PMN) were prepared by centrifuging isolated, intact PMN over a discontinuous Ficoll gradient that contained 20 microM cytochalasin B . The enucleated cells (PMN cytoplasts) contained about one-third of the plasma membrane and about one-half of the cytoplasm present in intact PMN . The PMN cytoplasts contained no nucleus and hardly any granules . The volume of the PMN cytoplasts was about one-fourth of that of the original PMN . Greater than 90% of the PMN cytoplasts had an "outside-out" topography of the plasma membrane . Cytoplasts prepared from resting PMN did not generate superoxide radicals (O2-) or hydrogen peroxide . PMN cytoplasts incubated with opsonized zymosan particles or phorbol-myristate acetate induced a respiratory burst that was qualitatively (O2 consumption, O2- and H2O2 generation) and quantitatively (per unit area of plasma membrane) comparable with that of intact, stimulated PMN . Moreover, at low ratios of bacteria/cells, PMN cytoplasts ingested opsonized Staphylococcus aureus bacteria as well as did intact PMN . At higher ratios, the cytoplasts phagocytosed less well . The killing of these bacteria by PMN cytoplasts was slower than by intact cells . The chemotactic activity of PMN cytoplasts was very low . These results indicate that the PMN apparatus for phagocytosis, generation of bactericidal oxygen compounds, and killing of bacteria, as well as the mechanism for recognizing opsonins and activating PMN functions, are present in the plasma membrane and cytosol of these cells.

Immunopharmacology, 1983 Aug, 6(2), 97 - 106
Effect of indomethacin on human neutrophil chemiluminescence and microbicidal activity; Horan TD et al.; The action of indomethacin on the bactericidal processes and chemiluminescent activity of human neutrophils was examined . Indomethacin substantially reduced the ability of neutrophils to kill Staphylococcus aureus and Escherichia coli and impaired their ability to generate both native and luminol-dependent chemiluminescence (CL) . In contrast to these effects, indomethacin exerted an amplifying effect on lucigenin-sensitized phagocytically induced CL . The data suggests that indomethacin interferes with neutrophil myeloperoxidase-mediated CL and microbial killing, while substantially augmenting PMN superoxide-forming activity.

Eur J Clin Microbiol, 1983 Aug, 2(4), 321 - 6
Enterotoxigenicity of Staphylococcus aureus strains from clinical isolates; Mauff G et al.; The enterotoxigenicity of 208 strains of Staphylococcus aureus isolated from clinical specimens and four strains isolated from food was investigated . All strains were examined for production of enterotoxins A, B, C, D, and E in cellophane-over-agar cultures using a modified optimum-sensitivity-plate method . Strains from systemic infections were also examined for production of toxic-shock toxin . The overall frequency of enterotoxigenic strains among the clinical isolates was 42% . Enterotoxin A was the predominant enterotoxin found among positive strains (45/88) . Enterotoxins B to E were found in decreasing order of frequency . Twenty-three strains produced two enterotoxins and one strain three toxins . A significant association was observed between multi-resistance of strains (defined as resistance against penicillin and at least two other antibiotics) and enterotoxin B production . Strains from phage groups I and III produced predominantly enterotoxin A . Four of the five strains producing toxic shock toxin belonged to phage group I; one was not classifiable.

Thorac Cardiovasc Surg, 1983 Aug, 31(4), 239 - 42
Perioperative infection prophylaxis in vascular surgery--a randomized prospective study; Salzmann G; In 300 reconstructive arterial operations of the abdominal aorta and the lower limbs, a randomized prospective study of the effect of perioperative infection prophylaxis with antibiotics was performed . The evaluation was carried out with regard to the frequency of severe postoperative wound infections . As prophylactic antibiotic Cefuroxime was initially used, Cefotaxim was later administered . Cefotaxim has been proven to be the appropriate antibiotic against the special microbial spectrum of our clinic . Side effects or complications due to the antibiotics were not found . Wound infections of all grades of seriousness occurred in the group without infection prophylaxis in 15.1%, in the antibiotic group in 3% (p less than 0.001) . Serious wound infection endangering or involving the transplant were found in the group without prophylaxis in 7.2%, whereas in the antibiotic group in 1.5% . Infections of the vascular prosthesis were observed in 2.4% and 0.8% respectively . Most wound infections occurred in the groin . The most common infecting organisms were Staphylococcus aureus or epidermidis . A higher risk of wound infections due to old age or gangrene could not be observed . Although wound infections in peripheral vascular surgery are relatively rare, they have severe consequences . It is therefore advisable to administer perioperative infection prophylaxis which significantly reduces this complication, especially in patients with synthetic implants.

J Immunol Methods, 1983 Jul 29, 61(3), 335 - 8
A rabbit antibody inhibition test to detect the antigenic activity of allergen extracts; Di Berardino L et al.; A method for standardizing the antigenic activity of allergen extracts is proposed, making use of a rabbit antiserum . A dilution of the latter is selected which is inhibited by the allergen extract . The antigenic activity is then evaluated by radioimmunoassay with 125I-labeled protein A from Staphylococcus aureus as tracer.

Arch Biochem Biophys, 1983 Jul 15, 224(2), 543 - 54
Isolation and characterization of protease do from Escherichia coli, a large serine protease containing multiple subunits; Swamy KH et al.; A new cytoplasmic proteolytic enzyme in Escherichia coli, named protease Do, has been purified to near homogeneity . The enzyme is an endoprotease that degrades casein, denatured bovine serum albumin, and globin but shows little or no hydrolytic activity against insulin, growth hormone, native bovine serum albumin, or a variety of commonly used peptide substrates . The molecular size of the enzyme was large, and it could be isolated in different preparations in either of two forms . One showed a molecular weight of about 500,000 on gel filtration and a sedimentation coefficient of 15.9 S on sucrose gradient centrifugation . The other appeared to be about 300,000 and sedimented at 12.7 S . No interconversion between the two forms and no other difference in the properties was found . Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS) shows that both forms contain a major 54,000-dalton band and three additional minor polypeptides with molecular weights of 45,000, 44,000, and 42,000 . These minor polypeptides appear to result from autolytic degradation of the major protein as demonstrated by peptide mapping with Staphylococcus aureus V8 protease . Thus, protease Do appears to contain a single subunit of 54,000, and can exist either as a decamer or as a hexamer or pentamer . The enzyme is a serine protease . It is sensitive to diisopropyl fluorophosphate (DFP) but not to metal chelating agents, sulfhydryl blocking groups, certain chloromethyl ketones, or various peptide aldehyde inhibitors . The enzyme covalently binds {3H}DFP, and the labeled subunit was visualized on SDS-polyacrylamide gels by fluorography . When cells growing in rich broth enter stationary phase, the relative concentration of protease Do increases more than twofold.

J Biol Chem, 1983 Jul 10, 258(13), 8290 - 7
Complete amino acid sequence of an allosteric enzyme, T2 bacteriophage deoxycytidylate deaminase; Maley GF et al.; The amino acid sequence of deoxycytidylate deaminase isolated from T2 phage-infected Escherichia coli has been determined . The enzyme is a hexamer, consisting of identical polypeptide subunits, each composed of 188 amino acids with a calculated Mr = 20,560 . The primary structure was established by automatic Edman degradation of the intact carboxymethylated protein and of peptides derived from the protein by cleavage with cyanogen bromide, trypsin, chymotrypsin, the Staphylococcus aureus V8 protease, and 2-(2-nitrophenylsulfenyl)-3-methyl-3-bromoindolenine . Knowledge of the primary structure of deoxycytidylate deaminase should aid in determining the allosteric binding site of the negative effector, dTTP, recently reported (Maley, F., and Maley, G.F . (1982) J . Biol . Chem . 257, 11876-11878), and eventually that of the enzyme's positive regulator, dCTP, as well as its substrate . The deaminase has been crystallized through the use of polyethylene glycol; a scanning electron micrograph is presented.

Klin Monatsbl Augenheilkd, 1983 Jul, 183(1), 60 - 2
{Bacteriological findings in conjunctival smears}; Seibel W et al.; Between February and December 1981 a total of 2,053 conjunctival smears were taken on chocolate agar plates . Of these specimens, 1,720 were taken from clear, noninflamed eyes on the day preceding intraocular surgery; 202 (11.7%) of the smears were positive . The most common pathogens were gram-positive cocci (119 X) and Staphylococcus aureus (47 X) . In 333 cases the smears were taken from the anterior segments of eyes with manifest infection and inflammatory signs . In this group pathogens grew on 95 smears (26%), the most common being gram-positive cocci (92 X) and Staphylococcus aureus (32 X) . As more than one pathogen grew in 80 of the 95 smear cultures, a total of 174 pathogens were found in this group.

J Clin Microbiol, 1983 Jul, 18(1), 194 - 6
Susceptibility testing of multidrug-resistant Staphylococcus aureus with the Sceptor microdilution system; Blair HC et al.; The antimicrobial susceptibilities of Staphylococcus aureus isolates were concurrently determined by the Sceptor system (BBL Microbiology Systems, Cockeysville, Md.) and by the standard disk diffusion method . For the methicillin-resistant isolates, there was greater than 98% agreement between the two test results with penicillin G, erythromycin, clindamycin, tetracycline, gentamicin, and tobramycin . Major disagreements (susceptible by one method and resistant by the other) were 7% for methicillin, 13.5% for cephalothin, 3.5% for cefamandole, and 27% for amikacin . The major discrepancies for methicillin were eliminated by supplementing the inoculum broth with salt . For methicillin-susceptible isolates, agreement between the two methods was 96 to 100% for all antibiotics except amikacin.

Am J Trop Med Hyg, 1983 Jul, 32(4), 809 - 11
Tropical pyomyositis of the iliacus muscle in American Samoa; Schecter W et al.; Ten cases of pyomyositis of the iliacus muscle are described in otherwise healthy young Samoans . The correct diagnosis was frequently delayed . Staphylococcus aureus was the most common pathogen isolated from drained abscesses . Preference for hip flexion was seen in six patients, and localized pain or induration at the anterior superior iliac spine were found in seven patients . These clues may help expedite a correct diagnosis.

J Bone Joint Surg Am, 1983 Jul, 65(6), 824 - 8
Septic arthritis of the wrist; Rashkoff ES et al.; Twenty-eight patients (two of them children) with septic arthritis of twenty-nine wrists were treated with early surgical drainage, parenteral antibiotics, and early motion after surgical decompression . The etiology was trauma in seventeen patients, and Staphylococcus aureus was the organism that was most commonly recovered on culture . In twenty-two patients (twenty-three wrists) who were followed for six months to nine years there were no recurrences . The results were evaluated in terms of range of motion, grip strength, and subjective complaints of discomfort and disability . Of the ten wrists with a good or excellent result, all had had the arthrotomy within ten hours after diagnosis, and of the thirteen with a fair or poor result, surgery had been delayed for sixteen hours or longer . The long-term results deteriorated in direct proportion to increasing time until treatment and the number of procedures performed.

Infect Immun, 1983 Jul, 41(1), 185 - 9
Selenium and immune functions in humans; Arvilommi H et al.; Earlier animal experiments have shown that selenium depletion may decrease immune functions . In this human study, 40 volunteers from a population with low serum selenium concentrations were supplemented with selenium or placebo for 11 weeks . Blood samples were drawn at intervals for analysis of selenium status and immune function . At the end of the supplementation period, plasma selenium levels were 74 ng/ml in the placebo group and 169 ng/ml in the supplemented group . The improvement in selenium status was associated with a 57% increase in the activity of platelet glutathione peroxidase in the group supplemented with selenium, but there was no increase in the activity of this enzyme in the placebo-treated subjects . Immune function was measured in vitro by tests of lymphocyte and granulocyte activity . Intracellular killing of Staphylococcus aureus by granulocytes was slightly lower in the placebo group than in the selenium group at the end of the supplementation period (77.2 compared to 85.2%; P less than 0.05) . No significant changes were observed in phagocytosis, chemotactic factor generation, antibody or leukocyte migration inhibitory factor production by lymphocytes, or proliferative responses to phytohemagglutinin or concanavalin A . These results suggest that the selenium deficiency of the order found in Finland and some other areas of the world has little, if any, influence on the immune functions measured in this study.

Arch Surg, 1983 Jul, 118(7), 791 - 4
Postoperative toxic shock syndrome; Morrison VA et al.; Previously, most cases of the toxic shock syndrome (TSS) have been described in a menses-related clinical setting . Recently, however, cases have been described in nonmenses-related clinical settings, in particular, in association with postoperative wound infections . Three cases of TSS occurred as a complication of surgical procedures . Toxic shock syndrome complicated the postoperative course of one patient undergoing routine inguinal herniorrhaphy, to our knowledge the first reported case of this syndrome directly resulting in the patient's death in the postoperative setting . In another case, TSS, with recurrence, resulted from a Staphylococcus aureus wound infection after unilateral salpingo-oophorectomy, to our knowledge the first reported case of recurrence in a nonmenses-related setting.

Arch Pathol Lab Med, 1983 Jul, 107(7), 351 - 7
A variant of toxic shock syndrome . Clinical, microbiologic, and autopsy findings in a fatal case; Smith JH et al.; A 16-year-old boy had toxic shock syndrome (TSS); Staphylococcus aureus bacteremia developed 11 hours prior to his death, which was three days after onset of the illness . The isoelectric focusing pattern of the staphylococcal isolate differed from both non-TSS and classic TSS S aureus isolates . Anatomic findings suggest three pathogenetic mechanisms: (1) immune complex-associated pulmonary microangiitis and vasculitis in the skin and skeletal muscle; (2) parenchymal cell "microvesicular" fatty metamorphosis in the liver, myocardium, renal tubules, and pancreas, and (3) pancarditis.

Pediatr Med Chir, 1983 Jul-Aug, 5(4), 157 - 60
{Antibiotic treatment of lung disease in cystic fibrosis}; Biasini GC et al.; Patients suffering from chronic lung infections are a major problems in therapy of cystic fibrosis (CF) . Resistance frequently occurs with great rapidity among isolates of Staphylococcus aureus and Pseudomonal aeruginosa . The rational for the use of frequent antibiotics (reduction of bacterial count), and the principle to follow while using antibiotics in CF patients are discussed . Research on amikacin peak levels and its average levels in serum, saliva and sputum is presented.

Rev Infect Dis, 1983 Jul-Aug, 5 Suppl 3, S509 - 14
In vitro study of the combination of rifampin with oxacillin against Staphylococcus aureus; Van der Auwera P et al.; The rapid emergence of strains resistant to rifampin suggests that for the treatment of bacterial infections rifampin should probably be combined with another antistaphylococcal drug . The authors examine the interaction of rifampin and oxacillin in various concentrations in vitro using a dynamic biophotometric technique . While synergism was observed occasionally, most concentrations studied exhibited either antagonism or indifference . No antagonism was observed with rifampin-resistant strains . Rifampin-resistant strains often emerged when the drug was tested alone . These findings again illustrate the complex and often unpredictable effect of combining rifampin with beta-lactam antibiotics.

Rev Infect Dis, 1983 Jul-Aug, 5 Suppl 3, S495 - 501
Chronic staphylococcal osteomyelitis: treatment with regimens containing rifampin; Norden CW et al.; Experience with rifampin as a component of combination antibiotic therapy for chronic staphylococcal osteomyelitis was reviewed . These data included the preliminary results of a randomized, prospective trial of parenteral nafcillin alone vs . parenteral nafcillin plus oral rifampin; the results of use of rifampin in combination with other oral antibiotics for the treatment of chronic staphylococcal osteomyelitis; and experience with rifampin regimens at the Oregon Health Sciences University . A total of 28 patients are described in this report; 70% of those who received regimens including rifampin experienced an apparent cure of infection . Most patients whose infections were not cured had inadequate surgical therapy or were also infected with other bacteria . Staphylococcus aureus was eradicated from the sinus drainage of all but one patient . Although these results are encouraging, a larger number of patients and a longer follow-up period are required for an adequate evaluation of the place of rifampin in therapy for chronic staphylococcal osteomyelitis.

Rev Infect Dis, 1983 Jul-Aug, 5 Suppl 3, S481 - 90
Rifampin in experimental endocarditis due to Staphylococcus aureus in rabbits; Zak O et al.; Rifampin possesses unique activity against Staphylococcus aureus . It is the most active antistaphylococcal antibiotic currently available and has been shown to be particularly effective in eradicating S . aureus from abscess cavities in experimental infections . However, resistance develops rapidly in vitro and in vivo when large numbers of organisms (10(6)-10(7)) are present, and use of combination therapy has been recommended . The use of combination therapy is complicated by the finding that in vitro the addition of rifampin may reduce (antagonize) the bactericidal effect of the beta-lactam antibiotics . This study examines the in vivo effect of treatment with a beta-lactam agent (cloxacillin), rifampin, or the combination on the eradication of S . aureus from cardiac vegetations in experimental endocarditis . Five different dosage combinations of the beta-lactam agent and rifampin were administered for a three-day period, and an attempt was made to correlate peak serum bactericidal titers with results of quantitative cultures of vegetations after therapy . In two of five regimens the combination of rifampin and cloxacillin produced enhanced efficacy in vivo (synergism); in two regimens the effect was no greater than the effect of either drug alone (indifference), and in one regimen the combination was less effective than either single-drug regimen alone (antagonism) . Peak serum bactericidal titers often were predictive of the in vivo effect when high doses of cloxacillin were used but were not consistently predictive of in vivo results when rifampin was the agent responsible for the major therapeutic effect . Rifampin-resistant strains did not emerge in animals receiving combination therapy but were isolated from vegetations from several animals receiving rifampin alone.

Rev Infect Dis, 1983 Jul-Aug, 5 Suppl 3, S468 - 73
Penetration of antibiotics into human leukocytes and dermal suction blisters; Solberg CO et al.; Staphylococcus aureus phagocytized by leukocytes from healthy donors and from patients with chronic granulomatous disease were protected from the antibacterial effect of gentamicin . Considerable numbers of phagocytized bacteria remained viable after exposure for 20 hr to antibiotic concentrations that killed greater than 99% of extracellular bacteria in less than 4 hr . A higher proportion of intracellular bacteria were killed by rifampin; this finding indicated that rifampin penetrates better into the phagocytic vacuole than does gentamicin and/or is more active against phagocytized bacteria than is gentamicin . After oral administration of 450 mg of rifampin to three healthy volunteers, concentrations of the antibiotic in serum and skin blister fluid were measured . Concentrations in serum peaked within 3 hr of oral administration (mean peak level, 13.2 micrograms/ml) . Concentrations in blister fluid peaked between 6 hr and 9 hr (mean peak concentration, 2.7 micrograms/ml) . Between 9 hr and 12 hr, the concentrations of rifampin in serum and blister fluid were similar; later, levels in blister fluid were higher than those in serum . The mean elimination half-life of rifampin was 2.5 hr in serum and 6.0 hr in blister fluid.

J Gen Microbiol, 1983 Jul, 129 (Pt 7), 2251 - 6
Plasmid replication in a temperature-sensitive chromosome replication mutant of Staphylococcus aureus; Summers DK et al.; Replication of the antibiotic resistance plasmids pI258, pT10501 and pC221 has been investigated in a mutant of Staphylococcus aureus NCTC 8325, which is temperature-sensitive for the initiation of chromosome replication . Replication of pI258 stopped rapidly at the nonpermissive temperature, whilst replication of pT10501 and pC221 continued (although at a lower rate than in the wild-type) . It is proposed that the product of the mutant gene may be required directly for pI258 replication, but not for replication of pT10501 or pC221.

Rev Infect Dis, 1983 Jul-Aug, 5(4), 692 - 712
Anaerobic osteomyelitis of long bones; Templeton WC 3rd et al.; Nine cases of anaerobic osteomyelitis of long bones are reviewed, and data are tabulated for an additional 52 cases from the literature . Three major clinical presentations of this disease entity include an acute hematogenous form usually involving previously normal bones in younger patients; a chronic infection in which the anaerobic bacteria superinfect a fracture site already compromised by osteomyelitis due to aerobic organisms, usually Staphylococcus aureus; and an indolent infection at the site of indwelling prosthetic devices . This last form appears to be initiated at the time of surgery by the introduction of normal cutaneous anaerobic flora into the wound, but disease does not become apparent until months to years later . The responsible organisms, pathophysiology, and clinical presentations of these three forms are discussed; and the therapeutic necessity to remove all devitalized tissues, reestablish vascularity, graft bony defects, and combine this with long-term antibiotic therapy, preferably bactericidal against the causative organisms, is emphasized.

J Clin Immunol, 1983 Jul, 3(3), 287 - 91
Chronic granulomatous disease of childhood: a changing pattern?
Frayha HH, Biggar WD.
We have followed nine male patients with Chronic Granulomatous Disease at The Hospital for Sick Children, Toronto, since 1972 . The diagnosis was established in each case by the failure of neutrophils to reduce nitroblue tetrazolium dye and to kill Staphylococcus aureus normally in vitro . Bacterial infections began between 6 months and 14 years of age . In five of the nine patients, infections began after 4 years of age . The first significant infection in five patients was a liver abscess(es), and one patient each had lymphadenitis, pulmonary aspergillosis, a parapharyngeal abscess, and a draining inguinal incision following surgery . Following diagnosis, all patients were started on Trimethoprim-Sulfamethoxazole at a dose of 2 mg/kg/day of Trimethoprim . The patients have been followed for 50 patient-years . Five of nine patients have been free of infection during 16 years of observation . For the remaining four patients, there have been six infections during 34 years of observation . A possible infection-related death occurred in one patient . The patients reported here appear to differ from those in previous reports . They present later in life, often with a liver abscess . They have a low incidence of subsequent bacterial infections which may, in part, be due to Trimethoprim-Sulfamethoxazole prophylaxis . The patients with chronic granulomatous disease reported here appear to have a better prognosis than previously thought.

J Immunol, 1983 Jul, 131(1), 238 - 43
Distinction of two subtypes of human leukocyte interferon (IFN-alpha) on B cell activation . B cell proliferation by two subtypes of IFN-alpha; Harada H et al.; We examined the effect of interferon (IFN), with particular emphasis on the effects of the two subtypes of IFN-alpha (IFN-alpha A and IFN-alpha B) on the B cell proliferation induced by Staphylococcus aureus Cowan I bacterium (SpA Col) . An increase of SpA Col-induced proliferation was observed in the presence of 100 to 1000 U/ml of IFN-alpha, but a decrease of SpA Col-induced proliferation was observed in the presence of 1000 to 10,000 U/ml of IFN-beta . The two subtypes of IFN-alpha had different effects on cell proliferation; a significant enhancement was shown in the presence of 1000 to 10,000 U/ml of IFN-alpha A, but inhibition was shown in the presence of 1000 to 10,000 U/ml of IFN-alpha B . In the reconstitution test of the two subtypes of IFN-alpha, the boundary between enhancement and inhibition of SpA Col-induced proliferation was revealed when the proportion of IFN-alpha A and IFN-alpha B (IFN-alpha A:IFN-alpha B) ranged between 8:2 and 9:1 . Toward the SpA Col-induced responses, the above IFN were all found to act on B cells directly, independent of the presence of T cells . Proliferative responses by IFN-alpha and IFN-alpha A, however, were shown to be slightly dependent on the presence of monocytes . The lymphocyte proliferation induced by other mitogens (phytohemagglutinin, concanavalin A, pokeweed mitogen, and protein A of S . aureus) were all inhibited by the above IFN.

Pathology, 1983 Jul, 15(3), 235 - 40
Strain differentiation in methicillin-resistant Staphylococcus aureus; Vickery AM et al.; Three different systems were used to test 236 isolates of methicillin-resistant Staphylococcus aureus in an attempt to ascertain if more than one strain is responsible for the current problem of cross-infection by this organism in N.S.W . hospitals . The biochemical tests used were of little assistance . Phage typing, using the Basic International Set of typing phages at 100 x routine test dilution (RTD), provided evidence of the presence of several different strains . Phage type 83A/85/95/90/88 was the typing pattern of the predominant strain and the nest most frequent group was not typable . These results were often difficult to read . Five new phages were therefore isolated and found to be valuable as they produced easily identifiable patterns at RTD.

Can J Biochem Cell Biol, 1983 Jul, 61(7), 802 - 10
Identification and characterization of estrogen-responsive gene products in the liver of rainbow trout; Chen TT; Injection of male rainbow trout with estradiol induced the production of a major serum protein, vitellogenin (VG), of molecular weight (MW) 170 000 . This protein is immunoprecipitable by an antiserum raised against lipovitellin from the eggs . Labelling by incubation of liver cubes of estradiol-treated fish with {35S}methionine and analysis of immunospecific products showed incorporation into two polypeptides of MW 170 000 and 160 000 . The peptide patterns generated from these two polypeptides by Staphylococcus aureus V8 protease digestion were identical . Cytoplasmic poly(A)+ RNA of estradiol-treated male liver directs the synthesis of a polypeptide of 160 000 daltons in the reticulocyte cell-free protein-synthesizing system or in Xenopus oocytes . This polypeptide is chemically, immunologically, and electrophoretically identical to the authentic VG polypeptide of 160 000 daltons . Changes in mRNA populations in the liver of male fish following estradiol treatment were detected on gels and by hybridization . On methylmercuric hydroxide gels, a polyadenylated RNA species of about 6300 nucleotides was found in estradiol-treated fish which was absent from the control fish . Data from R omicron t analysis revealed the presence of at least two mRNA sequences of the high frequency class in the RNA of hormone-induced fish and their sizes were determined by Northern blot analysis to be 6300 and 1800 nucleotides, respectively . These observations are consistent with the induction of vitellogenins and at least another estrogen-responsive gene product by estradiol as reported in other egg-laying vertebrates.

Eur J Cell Biol, 1983 Jul, 31(1), 55 - 61
Identification and characterization of axopodial tubulins from Echinosphaerium nucleofilum; Little M et al.; Isolated microtubule protein from axopodia of the heliozoan Echinosphaerium nucleofilum, consisting of two major bands on SDS-polyacrylamide gel electrophoresis (SDS-PAGE), has been compared to axonemal and cytoplasmic tubulins from both animal and non-animal sources . The upper E . nucleofilum protein band migrated faster than the alpha-tubulins of bovine brain and sea anemone sperm tails but with approximately the same electrophoretic mobility as the axonemal alpha-tubulins of Tetrahymena pyriformis and the alga Chlorogonium elongatum and cytoplasmic alpha-tubulin from the slime mold Physarum polycephalum . The lower E . nucleofilum protein band, however, had a higher electrophoretic mobility than all the beta-tubulins which we have so far examined . It was, nevertheless, a true beta-tubulin as shown by its migration on two-dimensional gel electrophoresis and the general resemblance of its one- and two-dimensional peptide maps to those of other beta-tubulins . The Staphylococcus aureus protease cleavage pattern of the upper axopodial protein band was similar to those of other non-animal alpha-tubulins but quite different from those of the animal alpha-tubulins . In contrast, the two-dimensional tryptic peptide map of axopodial alpha-tubulin was distinct from all of them . For example, a characteristic constellation of peptides common to the peptide maps of the other alpha-tubulins was absent from that of E . nucleofilum . In contrast to Physarum and metazoan tubulins but similar to Tetrahymena tubulin, the axopodial alpha-tubulin had a more basic isoelectric point than the beta-subunit as shown by two dimensional gel electrophoresis . Some of the unusual characteristics of E . nucleofilum axopodial tubulin may not only reflect phylogenetic variation, but also the different functional requirements of axopodial microtubules.

Appl Environ Microbiol, 1983 Jul, 46(1), 90 - 7
Surface thermodynamics of bacterial adhesion; Absolom DR et al.; The adhesion of five strains of bacteria, i.e., Staphylococcus aureus (strain 049), Staphylococcus epidermidis (strain 047), Escherichia coli (strains 055 and 2627), and Listeria monocytogenes, to various polymeric surfaces was studied . The design of the experimental protocol was dictated by thermodynamic considerations . From the thermodynamic model for the adhesion of small particles from a suspension onto a solid substratum, it follows that the extent of adhesion is determined by the surface properties of all three phases involved, i.e., the surface tensions of the adhering particles, of the substrate, and of the suspending liquid medium . In essence, adhesion is more extensive to hydrophilic substrata (i.e., substrata of relatively high surface tension) than to hydrophobic substrata, when the surface tension of the bacteria is larger than that of the suspending medium . When the surface tension of the suspending liquid is larger than that of the bacteria, the opposite pattern of behavior prevails . Suspensions of bacteria at a concentration of 10(8) microorganisms per ml were brought into contact with several polymeric surfaces (Teflon, polyethylene, polystyrene, and acetal and sulfonated polystyrene) for 30 min at 20 degrees C . After rinsing, the number of bacteria adhering per unit surface area was determined by image analysis . The surface tension of the suspending medium . Hanks balanced salt solution, was modified through the addition of various amounts of dimethyl sulfoxide . It was found that the number of bacteria adhering per unit surface area correlates well with the thermodynamic predictions and that these data may be used to determine the surface tension of the different bacterial species . The surface tensions of the bacteria obtained in this fashion are in excellent agreement with those obtained by other methods.

J Bacteriol, 1983 Jul, 155(1), 381 - 90
Electron microscopy of frozen-hydrated bacteria; Dubochet J et al.; Amorphous, unstained, frozen-hydrated sections of bacteria provide a faithful high-resolution image of procaryotic cells . Conventional preparation artifacts due to fixation, staining, and dehydration are nonexistent . Freezing damage is avoided by using glucose as a cryoprotectant . Cutting damage on frozen material is severe, but sectioning artifacts, being always related to the cutting direction, can be systematically recognized and thus taken into consideration . Geometry and density distribution of the bacterial envelope can be resolved to about 3 nm . The following main features have been observed . In Escherichia coli the inner and outer membranes have an approximately uniform density profile . The distance between the two membranes is constant, ca . 33 nm . In Staphylococcus aureus the cell wall is ca . 40 nm wide . It is bordered on the cytoplasmic side by an asymmetric 5.5-nm-wide bilayer . The bacterial nucleoid, clearly visible with conventional preparation methods, appears in exponentially growing bacteria as an ill-defined central region with approximately the same density as the rest of the cytoplasm . It becomes more clearly visible when bacteria are in the stationary phase, plasmolysed, fixed, or stained . We confirm that "mesosomes," hitherto quite often considered to be essential organelles in all procaryotes, are artifacts . They appear in large numbers during osmium fixation.

Rev Infect Dis, 1983 Jul-Aug, 5 Suppl 3, S474 - 80
Short-term administration of rifampin in the prevention or eradication of infection due to foreign bodies; Tshefu K et al.; Short-term administration of rifampin was evaluated as a means of preventing or eradicating infection due to foreign bodies . Tissue cages were implanted into guinea pigs and subsequently infected with 10(3) colony-forming units of Staphylococcus aureus Wood 46 . Infection developed in all tissue cages . Rifampin was administered thereafter intraperitoneally at a dosage of 7.5 mg/kg every 12 hr for 48 hr, and the tissue-cage fluid was monitored for possible development of infection by quantitative bacteriologic methods for 15 days . In all cases rifampin prevented or eradicated tissue-cage infection if treatment was initiated either 3 hr before or less than or equal to 12 hr after inoculation of microorganisms but was ineffective if initiated greater than 12 hr after inoculation . In cases of failure of treatment, rifampin-resistant variants could be demonstrated . Rifampin seems to prevent or eradicate tissue-cage infection only if given early after bacterial inoculation.

J Antimicrob Chemother, 1983 Jul, 12 Suppl A, 269 - 73
Penetration of ceftazidime into human tissue in patients undergoing cardiac surgery; Adam D et al.; The penetration of ceftazidime into human heart muscle, skeletal muscle, skin, fat and bone was measured in 39 patients undergoing cardiac surgery . Mean serum levels of 70 mg/l were measured 1.5 h after dosing with mean levels of 27 mg/l after 4 h and 5.5 mg/l after 12 h . Tissue levels of ceftazidime were measured up to 280 min after dosing . These levels were high with mean corrected levels after 120 min of 20.0 mg/l in bone, 7.1 mg/l in skin (after 100 min), 9.2 mg/l in fat, 12.7 mg/l in heart muscle and 9.4 mg/l in skeletal muscle . Serum and tissue levels were sufficient to prevent infection by all likely pathogens, including Staphylococcus aureus, 98% of which have a ceftazidime minimum inhibitory concentration of 8 mg/l or less . There were no post-surgical infections.

Infection, 1983 Jul-Aug, 11(4), 181 - 8
Toxic-shock syndrome: a commentary and review of the characteristics of Staphylococcus aureus strains; Chesney PJ; Toxic-shock syndrome (TSS) was first identified in 1978 in children as a toxin-mediated illness associated with Staphylococcus aureus . Extensive publicity ensued when the strong association of this disease with menses and tampon use was recognized in 1980 . Since that time, investigation into the epidemiology, clinical manifestations and pathogenesis of TSS has coincided with a decreasing incidence of disease . A unique exotoxin has been isolated under varying physico-chemical conditions from TSS-associated strains of S . aureus and its effect on immune function and other parameters both in vitro and in vivo described . In addition, TSS-associated S . aureus strains have been found to have characteristic heavy metal susceptibility patterns, to be lysogenized and to show decreased hemolysis on sheep blood agar . Preliminary results of the prevalence of TSS-associated strains of S . aureus and of the theories of pathogenesis are reviewed.

Am J Vet Res, 1983 Jul, 44(7), 1224 - 8
Depression of polymorphonuclear leukocyte function associated with experimentally induced Escherichia coli mastitis in sows; Lofstedt J et al.; In a study of susceptibilities of sows from 2 herds to experimentally induced Escherichia coli mastitis, a marked difference was seen . The "susceptible" sows were from a conventional herd and "resistant" sows were from a specific-pathogen-free herd . The purpose of the study was to determine whether deficient neutrophil function was associated with increased susceptibility to E coli-induced mastitis . Four in vitro procedures were used to evaluate polymorphonuclear leukocyte (PMN) function: (i) random migration under agarose, (ii) ingestion of 125I-iododeoxyuridine-labeled Staphylococcus aureus, (iii) quantitative nitroblue tetrazolium reduction, and (iv) iodination . After parturition and intramammary inoculation with E coli, sows from the susceptible herd were neutropenic and the neutrophils which were present in the peripheral blood had reduced function . Specifically, there were depressed random migration under agarose, S aureus ingestion, and iodination when compared with PMN function in resistant sows . These data indicate that susceptibility to E coli mastitis was associated with deficiencies in PMN numbers and function . Potential causes of the neutrophil dysfunction are discussed and include possible systemic hormonal aberrations or the presence of an inapparent viral or bacterial infection.

Diabetes, 1983 Jul, 32(7), 622 - 6
Diminished production of thromboxane B2 and prostaglandin E by stimulated polymorphonuclear leukocytes from insulin-treated diabetic subjects; Qvist R et al.; As an initial step to investigate the possibility that abnormal polymorphonuclear leukocyte (PMNL) function in diabetes might be related to abnormalities of arachidonic acid metabolism, product of the cyclooxygenase pathway were assayed in PMNL from 27 insulin-treated diabetic subjects and 27 age- and sex-matched nondiabetic subjects . It was found that the major prostanoid products formed were thromboxane B2 (TxB2) and prostaglandin E (PGE) . Production of both these substances was greatly enhanced in PMNL from control and diabetic subjects by exposure to a killed preparation of Staphylococcus aureus (S . aureus) or to zymosan . There was a marked reduction in the production of TXB2 by PMNL from diabetic subjects in response to stimulation by both S . aureus {670 +/- 98 (SE) versus 1010 +/- 76 pg/10(6) PMNL/90 min, P less than 0.01} and zymosan (583 +/- 53 versus 1034 +/- 46 pg/10(6) PMNL/90 min, P less than 0.001) . Similarly, production of PGE was significantly reduced in diabetics in response to both S . aureus (145 +/- 29 versus 232 +/- 16 pg/10(6) PMNL/90 min, P less than 0.05) and zymosan (181 +/- 21 versus 271 +/- 27 pg/10(6) PMNL/90 min, P less than 0.01) . There was no relation between the plasma glucose at the time of the test and the production of either prostanoid . Diminished production of cyclooxygenase products of arachidonic acid metabolism should be added to the known abnormalities of PMNL in diabetes . In view of the demonstrated or inferred effects of cyclooxygenase products on aspects of PMNL function, this observation may be important in understanding the pathogenesis of PMNL dysfunction in diabetes.

Eur J Biochem, 1983 Jul 1, 133(3), 515 - 21
Kinetic studies of aminoglycoside acetyltransferase and phosphotransferase from Staphylococcus aureus RPAL . Relationship between the two activities; Martel A et al.; In the Staphylococcus aureus strain harbouring the plasmid RPAL, the resistance to aminoglycoside antibiotics results from two inactivating reactions catalyzed by a 6'-N-aminoglycoside acetyltransferase and a 2"-O-amino-glycoside phosphotransferase . These enzymes are copurified with a constant ratio between the two activities, the purification process consisting in affinity chromatography, native electrophoresis and gel exclusion chromatography . The kinetic mechanisms of each activity have been determined from studies of initial velocities, as well as product and dead-end inhibitions . Both activities follow a random rapid equilibrium mechanism . The substrates and cofactors of one reaction have been tested as effectors of the other reaction . No interaction between the two activities has been observed . However, the GTP cofactor of phosphotransferase protects, at weak concentrations, the acetyltransferase against thermal inactivation, which suggests that the two activities may be associated.

Blut, 1983 Jul, 47(1), 21 - 30
Partial myeloperoxidase deficiency in preleukemia; Cech P et al.; In seven subjects with partial and apparently acquired form of myeloperoxidase (MPO) deficiency, some functional properties of neutrophils (PMNs) were studied . Five patients suffered from preleukemia, one from diabetes mellitus and one from carcinoma of the breast with bone marrow metastases . Intracellular bactericidal activity, oxygen consumption and superoxide radical production were within normal limits . In three patients with preleukemia, the serum opsonic activity was markedly reduced (less than m-3SD) in an autologous system, but normal in the presence of pooled normal serum . Decreased opsonic activity was also found when these patient's sera were assayed in the presence of normal PMNs . Since the levels of IgG and C3 were comparable in the patients' sera and the pooled serum, a deficiency of another unknown opsonin or the presence of an opsonization inhibitor has to be postulated . The partial MPO defect apparently doesn't decrease the intracellular killing of Staphylococcus aureus by PMNs . The known susceptibility to bacterial infections in preleukemia may be explained by the reduction of serum opsonization conducing to a secondary decrease of the ingestion and killing of bacteria by the PMNs.

G Batteriol Virol Immunol, 1983 Jul-Dec, 76(7-12), 218 - 26
{Phage-typing in the surveillance of hospital infections: tests on strains of Staphylococcus aureus}; Moiraghi Ruggenini A et al.; The sensitivity of 44 coagulase positive Staphylococcus aureus strains to antibiotics has been determined . They have been isolated from in-patients and staff in a piedmontese hospital . It has been possible to obtain the phage typing of 26 of these strains: 4 have been attributed to group I, 3 to group II, 13 to group III and 6 to the "miscellanea" . Some correlations have been established considering the data obtained by the laboratory and through epidemiologic research.

J Antimicrob Chemother, 1983 Jul, 12 Suppl A, 199 - 205
Ceftazidime in septicaemia; Lundbergh P et al.; Thirty-five patients with mean age 70 years (range 22 to 92) with 37 proven episodes of septicaemia (26) or bacteraemia (11) were treated with a mean ceftazidime dose of 1.80 (range 1 to 2 g) bid for 12 (range 3 to 17) days . Twenty patients had underlying diseases . Escherichia coli was the most common pathogen (19/37) . Of the 37 strains 36 were tested for antibiotic sensitivity and 35 were fully sensitive with MIC less than or equal to 8 mg/l (21 less than or equal to 0.12 mg/l) . The remaining strain, a Staphylococcus aureus, was moderately susceptible (MIC 16 mg/l) . Thirty patients (81%) were cured, six improved and one considered not evaluable . Serum concentrations were studied in 12 patients with slightly or moderately impaired renal function . They had trough and peak values ranging from less than or equal to 1.0 to 118.4 mg/l and 30 to 246.4 mg/l, respectively . In three patients the initial dose of 2 g bid had to be reduced to 1 g bid due to very high trough levels, and due to low peak value in one patient the dose was increased from 1 g to 2 g bid . Adverse reactions were mostly mild (positive Coombs' reaction, transiently raised aminotransferases or raised serum creatinine) . Treatment had to be stopped in two cases due to drug fever and/or rash . Therapy with ceftazidime 2 g bid seems to be successful in Gram-negative septicaemia; however, in patients with decreased renal function serum concentrations should be determined.

Clin Pediatr (Phila), 1983 Jul, 22(7), 524 - 5
Pseudo-toxic-shock syndrome due to a drug reaction; Bernstein DI et al.; A case is reported of a 13-year-old girl who had an illness initially thought to be toxic shock syndrome but subsequently noted to be a toxic reaction to the anticonvulsant, carbamazepine . The patient was never hypotensive but she was febrile and had a desquamative rash and involvement of three organ systems . Staphylococcus aureus was recovered from the patient's blood and vagina.

Am J Reprod Immunol, 1983 Jul-Aug, 4(1), 5 - 10
Suppression of the mitogen response of normal lymphocytes by serum from patients with testicular germ cell tumors; von Eyben FE et al.; Serum from seven of 15 patients with nonseminomatous testicular germ cell tumors impaired the transformation of lymphocytes from normal donors by phytohemagglutinin to less than 50% of that with addition to the cultures of normal AB serum . Similarly, serum from eight patients impaired the transformation by Staphylococcus aureus Cowan I . The impairment of the lymphocyte transformation did not correlate with the serum concentrations of alpha-fetoprotein (S-AFP) and human chorionic gonadotropin (S-HCG) . The patients with metastases and serum that impaired lymphocyte transformation to less than 50% of that with addition of serum from the controls and the other patients survived in the same way (0.50 less than p less than 0.75, log-rank test) . The prognosis of patients with testicular germ cell tumors seems not be influenced by S-AFP and S-HCG through an impact on lymphocyte functions determined as the response to the two mitogens.

Appl Environ Microbiol, 1983 Jul, 46(1), 283 - 5
Rapid isolation of DNA from Staphylococcus aureus; Dyer DW et al.; We describe a Staphylococcus aureus bulk DNA isolation procedure which uses detergent and guanidine hydrochloride to free the nucleic acid from contaminants . The procedure is rapid and yields high-molecular-weight DNA suitable for molecular biological procedures.

Arch Biochem Biophys, 1983 Jul 1, 224(1), 224 - 34
Multiple molecular forms of the gibberellin-induced alpha-amylase from the aleurone layers of barley seeds; Callis J et al.; A class of plant growth regulators, gibberellins, induce the synthesis of alpha-amylase (1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1) in the aleurone layers of barley (Hordeum vulgare L . var . Himalaya) seeds . The purified alpha-amylase is composed of multiple isozymic forms with indistinguishable molecular weights, but different net charges . These alpha-amylase isozymes separate on isoelectric focusing gels into two groups, each containing multiple species . One group has an apparent isoelectric point (pI) of approximately 5.8 (the high pI group) . The other group's pI values are around 4.5 (the low pI group) . On some gels a small amount of protein focuses between the high and low pI isozymes . These proteins comigrate with the low pI isozymes upon reelectrophoresis . The synthesis of these two groups is temporally regulated . The high pI group is the dominant set of isozymes secreted from embryoless half seeds during the first two days of gibberellin administration . After four days, however, the major isozymes are those of the low pI group . This shift in isozyme pattern is due to a shift in their relative rates of synthesis . Peptide analysis of these two groups of isozymes with Staphylococcus aureus V8 protease and cyanogen bromide shows amino acid sequence differences . However, members within the same group have similar peptide patterns . Both groups of isozymes are synthesized in vitro in a wheat germ extract primed with poly(A)+ RNA isolated from gibberellin-treated aleurone layers . This indicates that the synthesis of the two groups of alpha-amylase isozymes is probably directed by two or more different populations of mature mRNA . A model that explains these observations and the available genetic information is that barley aleurone alpha-amylase isozymes are encoded by at least two sets of structural genes.

J Biol Chem, 1983 Jun 25, 258(12), 7788 - 92
Identification of lactaldehyde dehydrogenase and glycolaldehyde dehydrogenase as functions of the same protein in Escherichia coli; Caballero E et al.; Lactaldehyde dehydrogenase is an enzyme involved in the aerobic metabolism of fucose in wild type Escherichia coli, and glycolaldehyde dehydrogenase is an enzyme involved in the metabolism of ethylene glycol in mutant cells able to utilize this glycol . Both enzyme sources display oxidative activity on either substrate with a constant ratio between these activities . We have found that both enzymatic activities present the same electrophoretic mobility when crude extracts were electrophoresed in polyacrylamide gels and the gels stained for enzyme activities . Furthermore, both enzymatic activities co-chromatograph in a DEAE-Sephadex column . If lactaldehyde dehydrogenase of wild type cells is purified near homogeneity and the purification procedure is screened for both aldehydes as substrates, only one enzyme is apparent, giving again a constant ratio between lactaldehyde and glycolaldehyde dehydrogenase activities . Genetic evidence of the fact that both activities are functions of the same protein is provided by the observation that mutation to thermosensitivity for the production of lactaldehyde dehydrogenase affected in the same way the production of glycolaldehyde dehydrogenase . Glycolaldehyde dehydrogenase from mutant cells is purified in a procedure coincident with the lactaldehyde dehydrogenase purification, yielding a single enzyme electrophoretically indistinguishable from the purified lactaldehyde dehydrogenase . Peptide mapping of the purified preparation after digestion with chymotrypsin or Staphylococcus aureus protease V8 gives an indistinguishable band pattern between both enzymes.

Carbohydr Res, 1983 Jun 16, 117, 113 - 23
The repeating sequence of the capsular polysaccharide of Staphylococcus aureus M; Murthy SV et al.; The anomeric configuration of the sugar residues of the capsular polysaccharide antigen of Staphylococcus aureus M were established by 13C-n.m.r . spectroscopy, and the linkage positions by g.l.c.-m.s . after methylation, indicating a leads to 4)-O-(2-acetamido-2-deoxy-alpha-D-galactopyranosyluronic acid)-(1 leads to 4)-O-(2-acetamido-2-deoxy-alpha-D-galactopyranosyluronic acid)-(1 leads to 3)-O-(2-acetamido-2-deoxy-alpha-D-fucopyranosyl)-(1 leads to repeating unit . A taurine residue is linked by an amide bond, on the average, to every fourth 2-acetamido-2-deoxy-D-galactopyranosyluronic acid residue.

Biochem Biophys Res Commun, 1983 Jun 15, 113(2), 611 - 7
Polyphemin: a teichoic acid-binding lectin from the horseshoe crab, Limulus Polyphemus; Brandin ER et al.; A Staphylococcus aureus-agglutinating lectin, capable of binding to N-acetyl-D-glucosamine, was isolated from the serum of Limulus polyphemus . The monosaccharide alone was incapable of inhibiting bacterial agglutination by this lectin . Quantitative precipitation studies with purified cell wall-derived teichoic acids, either devoid of or containing N-acetyl-D-glucosamine, confirmed the carbohydrate-binding specificity of the lectin and suggested that secondary, non-specific interactions contribute to binding biomolecules containing this sugar . The agglutination pattern with various S . aureus strains having N-acetyl-D-glucosamine-associated teichoic acid, teichoic acid without this sugar, and no teichoic acid indicated that this cell wall component is not the sole binding site for the lectin on intact S . aureus cells . Affinity gel chromatography, using N-acetyl-D-glucosamine-associated teichoic acid as the specific absorbent, has been used to isolate this lectin from Limulus serum.

Can Med Assoc J, 1983 Jun 15, 128(12), 1418 - 20
Staphylococcus aureus meningitis: 26 years' experience at Vancouver General Hospital; Roberts FJ et al.; The records of all patients with Staphylococcus aureus meningitis admitted to Vancouver General Hospital between 1956 and 1981 were reviewed . All the patients had clinical and laboratory features of meningitis, and in all cases S . aureus was isolated from the cerebrospinal fluid . S . aureus was responsible for 21 (3%) of the 710 cases of acute bacterial meningitis . Therapy with cloxacillin or methicillin, or both, with or without other agents, was successful in 14 of the 21 patients . Three of the 14 patients without ventricular shunts died, 2 with fulminating septicemia and 1 with a postoperative brain abscess treated with cloxacillin . Following shunt removal and antibiotic therapy all seven patients with ventricular shunts survived the infection . Shunt removal may therefore be essential in appropriate cases.

J Biol Chem, 1983 Jun 10, 258(11), 7175 - 80
Synthesis and secretion of apolipoprotein A1 by chick breast muscle; Shackelford JE et al.; The present work shows that chick breast muscles synthesize and secrete a protein which is very similar to chicken serum apolipoprotein A1 (Apo-A1), the major protein constituent of serum "high density" lipoprotein particles . This conclusion is based on the following observations . 1) When chick breast muscle explants were incubated in the presence of radioactive amino acids, a labeled protein of the same size as serum Apo-A1 (Mr approximately equal to 27,000) accumulated in the incubation media; 2) the muscle-derived secretory protein and serum Apo-A1 generated the same size distribution of peptide fragments following digestion with Staphylococcus aureus V8 protease; and 3) antibodies raised against serum Apo-A1 specifically precipitated the radioactive muscle secretory protein . The newly secreted muscle-derived Apo-A1 was associated with lipid, as judged by its "flotation" behavior during centrifugation of the labeled incubation media in the presence of 0.2 g/ml of sodium bromide; this observation suggests that muscle explants secreted Apo-A1 molecules as part of lipoprotein particles or that these Apo-A1 molecules became associated with lipid shortly after their secretion . The present work, together with the very recent report by Blue et al . (Blue, M.L., Ostapchuk, P., Gordon, J.S., and Williams, D.L . (1982) J . Biol . Chem . 257, 11151-11159) demonstrate that avian tissues other than liver and intestinal mucosa synthesize and secrete Apo-A1 . Results of short term amino acid incorporation experiments showed that chick breast muscles synthesize Apo-A1 at high rates only during the terminal stages of embryonic development and early stages of postembryonic maturation . Around the time of hatching, the relative rate of synthesis of Apo-A1 by chick breast muscle was found to be higher than in liver, a documented major site of synthesis of this apolipoprotein . Possible physiological implications of the present work will be considered.

J Biol Chem, 1983 Jun 10, 258(11), 7059 - 63
The regulatory components of adenylate cyclase and transducin . A family of structurally homologous guanine nucleotide-binding proteins; Manning DR et al.; G/F and transducin are guanine nucleotide-binding regulatory proteins that mediate activation of adenylate cyclase and of a rod outer segment cyclic GMP-specific phosphodiesterase, respectively . The substrate for islet-activating protein is a third guanine nucleotide-binding protein that is postulated to mediate inhibition of adenylate cyclase . The extent of structural homology among subunits of all three proteins was examined by analyses of amino acid compositions and electrophoretic patterns of proteolytic peptides . The lower molecular weight subunits (beta subunits; Mr = 35,000) of these proteins have identical amino acid compositions and yield similar peptides upon proteolysis with Staphylococcus aureus V8 protease and elastase . The higher molecular weight subunits (alpha subunits; Mr = 39,000, 41,000, and 45,000) are also similar to each other in these respects . Similarity between the subunits of transducin and the islet-activating protein (IAP) substrate is especially evident . Substantial differences do, however, exist between the lower and higher molecular weight subunits within each protein . In addition, evidence was obtained that the 41,000-Da subunit of the IAP substrate is not derived from the 45,000-Da subunit of G/F . It is concluded that transducin, the IAP substrate, and G/F represent a family of structurally homologous guanine nucleotide-binding regulatory proteins.

Dtsch Med Wochenschr, 1983 Jun 10, 108(23), 905 - 8
{The toxic shock syndrome in male patients}; Breuer N et al.; The toxic shock syndrome was observed in two young men (16 and 22 years) . In one it occurred after a purulent scraping wound of the skin, in the other two days after an operation with seemingly normal wound conditions . All findings returned to normal in both patients after about five weeks . The syndrome of toxic shock can occur, regardless of age and sex, in Staphylococcus aureus infections of various kinds.

J Rheumatol, 1983 Jun, 10(3), 503 - 6
Staphylococcal septic arthritis presenting as acute flare of pseudogout: clinical, pathological and arthroscopic findings with a review of the literature; Lurie DP et al.; A patient with primary hyperparathyroidism and known pseudogout presented with an acute flare of the right knee . Gram stain was negative, and many intracellular calcium pyrophosphate dihydrate (CPPD) crystals were seen by both compensated polarized light and transmission electron microscopy . Cultures grew Staphylococcus aureus; the joint was probably seeded by an antecedent skin infection, with enzymatic "strip mining" precipitating acute pseudogout . The patient was refractory to therapy with oxacillin, naproxen, intravenous colchicine, and closed drainage . Arthroscopic debridement with insertion of drainage tubes led to rapid improvement, and offers an alternative to arthrotomy in septic knee arthritis unresponsive to closed drainage.

Hoppe Seylers Z Physiol Chem, 1983 Jun, 364(6), 713 - 47
{Three-dimensional structure determination of antibodies . Primary structure of crystallized monoclonal immunoglobulin IgG1 KOL, I}; Schmidt WE et al.; The crystallizable myeloma immunoglobulin IgG1 KOL {allotype Gm(-1,4), (gamma 1, gamma)2} which is well characterized in its three-dimensional structure by X-ray diffraction analysis of high resolution has been proved to be homogenous by polyacrylamide gel electrophoresis . The H- and L-chains were separated by gel filtration after complete reduction and carboxymethylation and were characterized by amino acid analysis, end group determination and polyacrylamide gel electrophoresis, respectively . The intact IgG1 KOL was cleaved by cyanogen bromide and all CNBr-fragments were isolated and characterized . The reduced and carboxymethylated H-chain was digested by trypsin and the tryptic hydrolysate was separated by ion-exchange chromatography . Using different procedures of rechromatography 35 out of 37 tryptic H-chain peptides could be isolated in sufficient amounts, the missing 2 peptides were produced by tryptic digestion of 2 CNBr-fragments . The amino acid sequences of all tryptic peptides were determined using a modified Edman degradation method after separation of the enzymatic cleavage products by high-performance liquid chromatography (HPLC) . The complete primary structure of the VH-part of the H-chain was established by isolation and partial sequence determination of overlapping peptides obtained from cleavage of the intact H-chain by Staphylococcus aureus proteinase . The gamma 1-H-chain KOL comprises 455 amino acid residues and belongs to subgroup III . The switch from the variable to the constant part occurs at position 126/127, thus making VH-KOL one of the longest variable parts among the yet known immunoglobulin H-chains . This is due to the hypervariable region Hhv4 which is made up by 17 amino acid residues (4-9 residues more compared with other VH-parts) . Within this region a so far not described additional intrapeptidal disulfide bridge could be localized (Cys 105-Cys 110) that creates a short loop with antiparallel running peptide strains in beta-pleated sheet conformation . Its role in the three-dimensional structure of the antigen-binding site of the IgG1 KOL molecule is discussed using the data obtained from X-ray diffraction analysis.

Lab Anim Sci, 1983 Jun, 33(3), 258 - 63
Nasal dermatitis in the Mongolian gerbil; Bresnahan JF et al.; The prevalence of nasal dermatitis in two breeding colonies of Mongolian gerbils was determined; 56 of 302 (18.5%) were affected in Colony A, and 2 of 54 (3.7%) were affected in Colony B . The presence of nasal lesions did not correlate with sex, tail barbering, or epilepsy . There was a positive correlation between presence of nasal lesions and forepaw dermatitis, periocular alopecia, and protrusion of the nictitating membrane . Lesions were most common in the superior labial and lateral nasal regions . Histopathologic changes in the nasal area included acanthosis, mild hyperkeratosis, and inflammation of the dermis . Isolates of beta-hemolytic Staphylococcus aureus of several phage types were cultured from the nasal area of affected and nonaffected animals in both colonies . Staphylococcus saprophyticus isolates were associated with the development of early nasal lesions.

Am J Vet Res, 1983 Jun, 44(6), 986 - 8
Frequency and immunologic cross-reactivity of encapsulated Staphylococcus aureus in bovine milk in New York; Opdebeeck JP et al.; A total of 213 milk samples, which were positive by bacteriologic cultural examination for Staphylococcus aureus, were obtained from lactating cows on 55 farms throughout the dairying areas of New York . The S aureus isolates were evaluated for encapsulation directly from milk by the serum-soft agar technique . Results indicated that 93% of S aureus field isolates showed evidence of encapsulation on direct evaluation from milk . Suppression of capsule formation in vitro was seen in 72.7% of isolates when grown in the presence of 1% of anticapsular sera . One of the 3 anticapsular sera used exhibited a significantly higher level of cross-reactivity with the field isolates than did the 2 other sera . Increasing the concentrations of the antisera to 2% and 4% in the serum-soft agar had no effect on the expression of capsule in those isolates which had shown no evidence of capsule suppression with the specific antisera at the 1% level . Limited subculture of randomly selected encapsulated isolates on artificial media resulted in a decrease in the percentage of encapsulation from 100 to 33.3.

J Reticuloendothel Soc, 1983 Jun, 33(6), 429 - 42
A morphological study of the role of phagocytes in the clearance of Staphylococcus aureus from the lung; Lipscomb MF et al.; A nonlethal dose of Staphylococcus aureus was inoculated into the mainstem bronchus of mice in order to study the influx of polymorphonuclear leukocytes (PMN) . The goal was to determine the routes of entry of PMN into the lung following bacterial challenge, the relative importance of PMN as compared to alveolar macrophages (AM) in the uptake of S aureus, and the role of lymphatics in clearance of intact microorganisms . Resident AM took up S aureus within minutes of inoculation, but PMN were subsequently recruited to the lung and were the predominant cell containing S aureus by 4 hours following inoculation . PMN were recruited from arteries, capillaries, and venules . Emigration of PMN into alveolar spaces occurred between type I epithelial cells as well as between type I and type II epithelial cells . Lymphatics played only a minor role in the clearance of S aureus.

J Infect Dis, 1983 Jun, 147(6), 1018 - 29
Peritoneal macrophages and opsonins: antibacterial defense in patients undergoing chronic peritoneal dialysis; Verbrugh HA et al.; The antibacterial activity of phagocytic cells and opsonins in peritoneal dialysis effluents from 21 patients undergoing chronic peritoneal dialysis (CPD) was studied . Effluents contained an average of 12 x 10(6) cells per liter that were predominantly macrophages . Macrophages phagocytized and killed opsonized Staphylococcus epidermidis, Staphylococcus aureus, and Escherichia coli as efficiently as did polymorphonuclear neutrophils (PMNs) from healthy donors . Macrophage chemiluminescence was one-third of that observed with donor PMNs . In the absence of opsonins, macrophages efficiently phagocytized and killed S . aureus by binding S . aureus cell wall protein A to macrophage surface IgG . Nine (43%) of 21 effluents failed to opsonize S . epidermidis, and none opsonized E . coli . When present, titers of S . epidermidis opsonins were 50- to 100-fold lower than that of normal serum . IgG and C3 concentrations in effluent reflected its opsonic capacity . Macrophages from patients undergoing CPD thus have intact phagocytic and bactericidal functions . However, the low level of opsonic molecules and inadequate numbers of macrophages in the peritoneal cavity may predispose patients undergoing CPD to peritonitis.

Environ Res, 1983 Jun, 31(1), 66 - 75
The effects of chronic tobacco smoke exposure from high-tar cigarettes on the phagocytic and killing capacity of polymorphonuclear cells of mice; Keast D et al.; Mice were exposed to high-tar (HT) (16 mg tar/cigarette) filtered cigarettes for up to 32 weeks . Intraperitoneal caseinate induction of polymorphonuclear (PMN) cells in both tobacco smoke (TS)-exposed and age-matched control mice provided the source of cells which enabled an assessment to be made in vitro of both their phagocytic and degradative properties . Radioactive Staphylococcus aureus were opsonized with either immune serum or heat-inactivated (complement deficient) immune serum for the tests . Two strains of mice were used in all trials . The PMN cells from BALB/c strain mice were less affected by TS exposure than those of C57 Black mice in both phagocytic and degradative capacities when the S . aureus were opsonized with immune serum . However, heat inactivation of complement within the serum reduced the differences between TS-exposed animals and age-matched controls in both strains of mice . The treatment effects were seen at all times tested from 3 days to 32 weeks of TS exposure . The results suggest that fc-receptor site activity of PMN cells is not significantly affected by TS exposure but that complement interactions within the phagocytic process are significantly suppressed . Whether this is due directly to complement receptors or metabolic associations of phagocytosis of the complement/opsonin/antigen complex is unclear.

Clin Sci (Lond), 1983 Jun, 64(6), 655 - 9
A radioimmunoassay for alpha- and beta-gliadins; Ciclitira PJ et al.; 1 . A rapid, sensitive specific radioimmunoassay for alpha- and beta-gliadin has been developed using an antiserum raised in rabbits to A-gliadin, a component of alpha-gliadin . 2 . The antigen used in the assay was alpha-gliadin labelled with 125I; antigen-antibody complexes were collected after adsorption to Staphylococcus aureus in suspension . 3 . The sensitivity of the assay, as judged by competitive binding with unlabelled antigen, was 1 ng of alpha- or beta-gliadin, which show complete cross-reaction with this antiserum . 4 . Cross-reactivity to other wheat proteins was less than 1% and no cross-reactivity to extracts of rye, barley or oats was observed . 5 . This radioimmunoassay for alpha- and beta-gliadin has been used to measure their amount in different varieties of wheat flour, several foods prepared from flour, e.g . bread, biscuits and products prepared as 'gluten free' . The possibility of assaying for alpha-gliadin in prepared foods is of special value since alpha-, beta-, gamma- and omega-gliadin have been shown to exacerbate coeliac disease.

Arch Microbiol, 1983 Jun, 134(3), 208 - 11
The effect of glucose on the differential rates of extracellular protein and alpha-toxin formation by Staphylococcus aureus (Wood 46); Coleman G; The differential rates of formation of total extracellular protein and alpha-toxin by Staphylococcus aureus (Wood 46) were determined during aerobic growth, at 37 degrees C, in a complex medium containing 0.0, 0.25 or 1.0% (wt/vol) glucose . Different inocula were employed from 1% (vol/vol) of an overnight culture to 100% where bacterial cells were washed and resuspended in fresh medium without change in density . It was shown that under all conditions examined the differential rates of total extracellular protein formation exhibited a biphasic pattern characteristic of regulation based on 'competition' . This biphasic pattern was maintained even in the presence of a large inoculum and a high glucose concentration, conditions considered to favour the onset of catabolite repression . However, a lowering of the initial rate was observed with increasing glucose suggesting the superimposition of catabolite repression as a modulating effect under extreme conditions . In the case of the specific extracellular protein component, alpha-toxin, its differential rate of formation paralleled total exoprotein in all except the condition most favourable for catabolite accumulation when a deviation consistent with a pronounced catabolite repression of this component was demonstrated which was not pH-dependent.

Antimicrob Agents Chemother, 1983 Jun, 23(6), 930 - 1
Effect of beta-lactam antibiotics on migration and bactericidal activity of human phagocytes; Fietta A et al.; The effect of beta-lactam antibiotics upon some functions of human phagocytes was examined . Penicillins (carbenicillin and piperacillin), thienamycin, and cephalosporins (cefotetan, ceftazidime, and moxalactam) had no effect on either the random or directional migration or on the bactericidal activity of neutrophils and monocytes against Staphylococcus aureus . On the contrary, cefoperazone at therapeutic levels was shown to inhibit neutrophil chemotaxis.

Antimicrob Agents Chemother, 1983 Jun, 23(6), 938 - 40
Mechanism of resistance to some cephalosporins in Staphylococcus aureus; Kono M et al.; The mechanism of resistance to some cephalosporins in Staphylococcus aureus strains was investigated with high-pressure liquid chromatography and nuclear magnetic resonance spectrometry . Drug inactivation by penicillinase was found to be the main mechanism of resistance to cefazolin, cephaloridine, and cephalothin in S . aureus.

Hepatogastroenterology, 1983 Jun, 30(3), 86 - 9
B lymphocyte function stimulated by staphylococcus aureus Cowan I in chronic liver disease; Kakumu S et al.; Peripheral blood B cell response to staphylococcus aureus (SpA) Cowan I was evaluated in 8 healthy subjects, 8 patients with liver cirrhosis (LC), and 2 patients with chronic active hepatitis (CAH) with hypergammaglobulinemia (greater than 2 g/dl) . In control studies it was shown that stimulation by SpA Cowan I was much less T cell-dependent than that induced by pokeweed mitogen (PWM) or con-canavalin A (ConA) when the amounts of immunoglobulins (Ig) secreted into culture supernatants were measured by radioimmunoassay (RIA) and the blastogenic response was measured by incorporation of tritiated thymidine . B cells from only 2 patients revealed increased Ig synthesis by SpA Cowan I stimulation . In the study of blastogenic response, increased DNA synthesis of B cells by SpA Cowan I stimulation was observed in 2 patients and decreased DNA synthesis in 3 patients . The remaining patients demonstrated normal range response . There was no correlation between B cell response to SpA Cowan I and clinical data such as gammaglobulin level in the patients studied . These studies indicate that B cell function remains intact in many patients with chronic liver disease with hypergammaglobulinemia.

J Infect Dis, 1983 Jun, 147(6), 1078 - 89
Staphylococcal beta-lactamase and efficacy of beta-lactam antibiotics: in vitro and in vivo evaluation; Chapman SW et al.; The susceptibility of five cephalosporins and nafcillin to changes in inoculum size of 26 isolates of Staphylococcus aureus was studied . The spectrum of antibiotic sensitivity to such changes was cefazolin = cephaloridine greater than cefamandole greater than cephalothin greater than cefoxitin = nafcillin . Half of the isolates resulted in large and half in minimal inoculum-induced changes in minimal inhibitory concentrations (MICs) . These changes correlated with the amount of beta-lactamase produced by the isolates . The in vivo relevance of these findings was studied in a model of intraperitoneal infection in mice . The effect of beta-lactamase production on mortality was greatest among animals given cefazolin or cephaloridine, intermediate among those given cefamandole, and nonexistent among those given cefoxitin, cephalothin, or nafcillin . The number of organisms in the animals' spleens paralleled survival rates . An increase in the serum level of cefazolin increased the survival rate among mice given that drug . Hence, the survival of mice was influenced by (1) the ability of the infecting organism to increase the MIC of an antibiotic via inoculum increases, (2) the sensitivity of the antibiotic to beta-lactamase, and (3) the peak level of antibiotic attained in serum.

Clin Pediatr (Phila), 1983 Jun, 22(6), 407 - 11
Bacterial tracheitis--an old disease rediscovered; Sofer S et al.; During a 3-year period, seven children with bacterial tracheitis were admitted to the intensive care unit of the Winnipeg Children's Hospital . The illness was characterized by fever, toxicity, and stridor . Respiratory difficulty was secondary to copious thick purulent tracheal secretions . In the majority of patients the illness was caused by Staphylococcus aureus, and the rest had Hemophilus influenzae infection . Viral studies in five patients were negative . Most patients required endotracheal intubation and frequent tracheal toilet to prevent serious airway obstruction . In our ICU, bacterial tracheitis accounted for about 14 per cent of admissions with infectious upper airway obstructive illness, while epiglottis and croup accounted for 55 per cent and 31 per cent, respectively . Only 5 per cent of children with croup admitted to the hospital were admitted to the ICU . Bacterial tracheitis has reappeared, at least in North America, as an important and serious cause of obstructive upper airway disease in children and must be recognized early in order to prevent catastrophic airway obstruction.

J Clin Microbiol, 1983 Jun, 17(6), 1170 - 2
Temperature-related variations in the lipid composition of methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains; Pisano MA et al.; Growth of methicillin-susceptible and -resistant strains of Staphylococcus aureus at 28 and 37 degrees C led to changes in the relative amounts of phospholipids recovered . The major phospholipids present in these strains were phosphatidylglycerol, lysophosphatidylglycerol, and diphosphatidylglycerol . Phosphatidylglycerol tended to occur in lower amounts in the resistant strains, but it was the major phospholipid in both methicillin-susceptible and -resistant S . aureus strains . Growth patterns and total lipid concentrations were similar in all strains.

Immunology, 1983 Jun, 49(2), 239 - 44
Association of human leucocyte low responsiveness to inducers of interferon alpha with HLA-DR 2; Abb J et al.; The influence of antigens of the major histocompatibility complex (MHC) on the production of interferon (IFN) alpha or IFN gamma by human peripheral blood leucocytes (PBL) in vitro has been studied . Synthesis of IFN gamma by PBL stimulated with purified phytohaemagglutinin (PHA-P) or protein A of Staphylococcus aureus (SpA) appeared not to be controlled by MHC antigens . The production of IFN alpha, however, was influenced by the HLA type of the donor . Low responsiveness of PBL to inducers of IFN alpha (influenza virus, Molt 4 cells) was associated with HLA-DR 2 . Implications of these observations for studies of IFN production and natural killer (NK) cell activity are discussed.

J Dairy Sci, 1983 Jun, 66(6), 1384 - 9
Novel immunofluorescent technique for identification of ovine immunoglobulins and other potential opsonins binding to live Staphylococcus aureus; Watson DL et al.; A qualitative in vitro technique has been developed for identification of potential opsonins (immunoglobulins G1, G2, A, M; the third component of ovine complement C3, and fibronectin) of sheep origin, binding to cell walls of viable Staphylococcus aureus . The technique uses monospecific antisera to these ovine proteins . The antisera are conjugated to FITC-protein A such that the protein A-binding sites in the Fc region of the immunoglobulin molecules are occupied with FITC-protein A complexes and are prevented, therefore, from binding to protein A in the staphylococcal cell wall . The technique is highly specific and sensitive, and once conjugated monospecific antisera are prepared, many tests can be done in a short time . Staphylococcus aureus incubated in samples of milk whey showed variable binding of immunoglobulins G1, G2, and M; immunoglobulin A was bound from only one sample of milk whey, and the third component of ovine complement C3- and fibronectin binding were not detected . Most samples of blood serum and colostral whey produced binding of immunoglobulins G1, G2, A, and M, and the third component of ovine complement C3 to the Staphylococcus aureus cell surface; fibronectin binding was found in serum but not in colostral whey . Washings from involuted mammary glands invariably produced binding of all immunoglobulins but not of the third component of ovine complement C3 or fibronectin.

Aust N Z J Surg, 1983 Jun, 53(3), 245 - 8
Postoperative wound infections: a study of bacteriology and pathogenesis; Kune GA et al.; Of 200 patients, 6.5% were skin carriers of Staphylococcus aureus and 3.5% became skin carriers after admission . Development of skin carrier state was associated with a long preoperative hospital stay . A second study of 275 cases showed that skin carriers of S . aureus had a significantly higher subsequent staph wound infection rate than non-carriers, but nose and/or throat carriers of S . aureus did not . Peroperative wound contamination was a significant factor in the subsequent development of wound infection . In clean surgery the infection rate was 5.9% . Both wound contamination and infection was due to gram positive organisms, usually S . aureus . When the gastrointestinal or biliary tract had been opened, the infection rate was 28%, the usual contaminating and infecting organisms were enteric and only occasionally S . aureus . There is a need for improved methods of minimizing peroperative wound contamination.

Pathol Biol (Paris), 1983 Jun, 31(6), 551 - 4
{Growth of Staphylococcus aureus Cowan I in the presence of subbactericidal concentrations of povidone-iodine}; Reverdy ME et al.; Staphylococcus aureus, strain Cowan I, is cultivated in the presence of povidone iodine (PVP.I) at the concentration of 0, 640, 960 micrograms/ml; growth curves points are platted (OD 640 mm) at 0, 1, 2.50, 3, 4.50, 7 and 24 hours . Growth curves in the presence of povidone alone as well as in the standard without antiseptic are similar and reproducible . A bacteriostatic effect is observed with 640 micrograms/ml PVP.I concentration compared to an incomplete bactericidal effect with 960 micrograms/ml concentration . Proteinee A detected by conditioned hemagglutination method is present in the centrifugation pellet of the standard and only in the supernatant of the test samples during the early phases of the growth . Production of alpha-ribitol-teichoic acid detected by counterimmunoelectrophoresis is not significantly different in the standard and in the presence of antiseptic . Biochemical characteristics studied by Api Staph gallery are identical for the control strain and the strain grown in the presence of PVP.I . Observed by scanning and transmission electronic microscope the bacterial cells do not present structural modification during growth with PVP.I.

Pathol Biol (Paris), 1983 Jun, 31(6), 548 - 50
{Additive effect of an immunomodulator on the therapeutic effect of antibiotics}; Gillissen G et al.; The effect of an immunomodulator, the Immunoferon (IF), on the course of experimental infections with Staphylococcus aureus and on cellular immune reactions (foot pad swelling test) was examined . The substance is a glycophosphopeptide adsorbed on sulfurized calcium phosphate . The IF improves clearly the therapeutic effect of penicillin, but has no significant effect when given alone . The substance neutralizes the negative effect of hydrocortisone on the therapeutic effectivity of penicillin . The cellular immune reaction is reduced by very small doses of IF, but is enhanced progressively by higher doses . The effect of hydrocortisone is abolished in this respect.

Pathol Biol (Paris), 1983 Jun, 31(6), 497 - 500
{Effect of trimethoprim at a subinhibitory concentration on the appearance of mutants resistant to rifampicin}; Joly B et al.; Experiments have been performed to establish the effect of trimethoprim (Tmp) on the emergence of Staphylococcus aureus (S . aureus) mutants to rifampicin (Ramp) . Four S . aureus strains were used: two strains with low MIC of Ramp (0,01 microgram/ml), two strains with enhanced MIC of Ramp (2 micrograms/ml) . For each of them the frequency of mutants and the mutation rates to Ramp were determined when various concentrations of Tmp were added . Results showed that subinhibitory concentrations of Tmp reduced 10 to 100 folds the frequency of mutants and the rates of mutation to Ramp alone . The more the Tmp concentrations were near the MIC, the more this effect was important.

J Clin Microbiol, 1983 Jun, 17(6), 1021 - 5
Rapid detection by a coagglutination test of heat-labile enterotoxin in cell lysates from blood agar-grown Escherichia coli; Ronnberg B et al.; Colonies of Escherichia coli from blood agar plates were suspended and lysed in saline containing polymyxin B and a detergent (Triton X-100) . The lysates were assayed for heat-labile enterotoxin (LT) by a coagglutination test (coa-test) . The coa-test reagent consisted of Formalin-treated and heat-killed cells of Staphylococcus aureus, strain Cowan 1, sensitized with a high-titer rabbit anti-LT serum . Purified LT was detected in the coa-test at the nanogram level (2 to 5 ng), whereas larger amounts of cholera toxin (50 ng) were required to give a positive test . The coa-test was compared with the CHO cell test for detection of LT among E . coli strains isolated from human and animal stool cultures . The results of the coa-test and the CHO cell test correlated with 63 of 67 strains of human origin . Six of nine animal strains, defined as LT positive by the CHO cell test, gave positive results in the coa-test . We conclude that the coa-test is probably accurate and sensitive enough to be used in routine diagnosis of LT-producing E . coli strains isolated from human stool cultures . No special laboratory equipment is required, which makes the coa-test suited for diagnosis of enterotoxigenic E . coli in small hospital laboratories and in developing countries.

J Natl Cancer Inst, 1983 Jun, 70(6), 1127 - 33
Protein A-independent tumoricidal responses in dogs after extracorporeal perfusion of plasma over Staphylococcus aureus; Gordon BR et al.; Protein A-positive or -negative Staphylococcus aureus preparations were used in an extracorporeal system to treat dogs with spontaneously occurring cancers . Tumor regression was seen in 4 of 7 dogs treated by reinfusion of plasma that had been incubated with protein A-positive S . aureus Cowan I strain (SAC) . Therapy was associated with fever, liver enzyme abnormalities, and hypocomplementemia . Tumor response and toxicity could be diminished by more extensive washing of the SAC preparation . Tumor regression was also seen in 2 of 2 animals treated with protein A-negative S . aureus Wood strain 46 . In addition, tumors regressed in 3 of 4 dogs treated with infusions of protein A-free saline extracts from S . aureus . These results suggest that the release of a non-protein A bacterial product contributes to tumor regression following incubation of plasma with S . aureus.

Antimicrob Agents Chemother, 1983 Jun, 23(6), 817 - 26
Analysis of plasmids in nosocomial strains of multiple-antibiotic-resistant Staphylococcus aureus; Lyon BR et al.; Nosocomial infections caused by Staphylococcus aureus strains resistant to methicillin and multiple antibiotics have reached epidemic proportions in Melbourne, Australia, over the past 5 years . Plasmid analysis of representative clinical isolates demonstrated the presence of three classes of plasmid DNA in most strains . Resistance to gentamicin, kanamycin, and tobramycin was usually mediated by an 18-megadalton plasmid but could also be encoded by a related 22-megadalton plasmid . Two distinguishable plasmids of 3 megadaltons each endowed resistance to chloramphenicol, and the third class consisted of small plasmids, each approximately 1 megadalton in size, with no attributable function . An extensive array of resistance determinants, including some which have usually been associated with a plasmid locus, were found to exist on the chromosome . Evidence that resistance to gentamicin, kanamycin, and tobramycin is chromosomally encoded in some clinical isolates suggests that this determinant may have undergone genetic translocation onto the staphylococcal chromosome.

Inflammation, 1983 Jun, 7(2), 145 - 53
Effects of leukocyte inhibitory factor (LIF) on human neutrophil function; Klempner MS et al.; The effects of the lymphokine, leukocyte inhibitory factor (LIF), on human neutrophil function were studied . This soluble mediator, which is defined by its specific inhibition of neutrophil locomotion, does not interfere with chemotactic factor binding and does not affect basal or stimulated superoxide generation by neutrophils . In contrast, phagocytosis of opsonized Staphylococcus aureus is markedly inhibited by LIF, and degranulation is stimulated by this lymphokine . The possible mechanisms of LIF action on neutrophils are discussed.

J Invest Dermatol, 1983 Jun, 80(6), 494 - 6
The Staphylococcus aureus receptor for fibronectin; Bibel DJ et al.; The Staphylococcus aureus binding site for human fibronectin was determined by unique biologic assays based upon the specific adherence of the bacterium to nasal epithelial cells . Fibronectin treatment of S . aureus caused a reduction in adherence tallies on high granular and fully keratinized cells compared to controls (p less than 0.05; p less than 0.001) . Spinous and low granular cells showed no significant differences . The cell wall materials N-acetyl-D-glucosamine, N-acetyl muramic acid, and protein A were unable to inhibit the coupling of fibronectin to S . aureus . Only ribitol teichoic acid had this property . Furthermore, fibronectin could neutralize the adherence-blocking ability of teichoic acid, which affects keratinized cells . Thus, teichoic acid seems to be a receptor for fibronectin.

Infect Immun, 1983 Jun, 40(3), 1004 - 10
Synovial fluid inhibits killing of Staphylococcus aureus by neutrophils; Simon GL et al.; Serum in the extracellular environment promotes neutrophil bactericidal activity apart from its opsonizing properties . We examined the effect of non-inflammatory osteoarthritic synovial fluid on serum-mediated neutrophil killing of Staphylococcus aureus . This was done to evaluate the effect of synovial fluid on neutrophil bactericidal activity independent of opsonin concentration . With an initial inoculum of 5 X 10(6) CFU/ml, 1.47 +/- 0.14% bacteria survived after 120 min of incubation with 10% serum and neutrophils . In contrast, 4.07 +/- 0.33% bacteria survived after incubation in serum plus synovial fluid (P less than 0.001) . This inhibitory effect was directly related to the concentration of synovial fluid in the incubation mixture . Increasing the concentration of synovial fluid resulted in an increased percent survival . Studies utilizing preopsonized bacteria and radiolabeled organisms demonstrated that synovial fluid did not interfere with opsonization or phagocytosis . Intracellular bactericidal activity was assayed separately from phagocytosis by utilizing a brief ingestion period followed by the removal of extracellular bacteria by either differential centrifugation or lysostaphin treatment . The reincubation of cells and associated bacteria with serum or serum plus synovial fluid revealed that synovial fluid significantly inhibited the promoting effect of serum on neutrophil bactericidal activity . After 60 min of incubation with 10% serum, 13.0 +/- 1.2% bacteria survived, whereas 21.5 +/- 2.3% survived after incubation in serum plus synovial fluid (P less than 0.005) . Superoxide production was not affected by the presence of synovial fluid . These findings suggest that the inhibitory effect of synovial fluid is due to an interaction between synovial fluid and the serum factors that promote intracellular killing.

Blood, 1983 Jun, 61(6), 1247 - 57
Human neutrophil dysfunction with giant granules and defective activation of the respiratory burst; Newburger PE et al.; We describe a patient whose peripheral blood neutrophils and bone marrow precursors (beyond promyelocytes) contained multiple large azurophilic granules . There were also giant granules in eosinophils, basophils, melanocytes, renal tubules, thyroid, and neurones, but not lymphocytes or monocytes . His clinical course included recurrent (ultimately fatal) infections and severe neurologic impairment . Immunofluorescent staining with fluoroscein- and rhodamine-conjugated antisera to primary and secondary granule markers showed virtually all of the granulocyte granules and rare monocyte granules to be fusion products containing both markers . Electron microscopy showed the granules to be large peroxidase-containing lysosomes . Only rare normal primary and secondary granules were present . Superoxide generation in response to opsonized zymosan was 7.3 nmole/min/10(6) cells (control 8.9); but in response to phorbol myristate acetate, only 2.2 (control 9.4) . Nitroblue tetrazolium slides showed 3+ dye reduction in response to opsonized zymosan by 90% of granulocytes (control 91%) and to phorbol myristate acetate by 22% (control 99%), with 71% producing only a minimal 1+ response . Cellular contents of myeloperoxidase and beta-glucuronidase were elevated, but the percent release during exocytic degranulation was equivalent to control . Ingestion of complement-opsonized Staphylococcus aureus and zymosan was also normal . Killing of Staphylococcus aureus was 60% at 90-min incubation (control 92%) . Granulocyte cyclic adenosine monophosphate (AMP) content was 4 pmole/10(7) cells (control 3.1) . In order to determine whether these characteristics derived from the cells' genetic program or their environment, the patient's bone marrow was grown in long-term culture . Granulocytes produced in vitro demonstrated the same morphology, same defect in activation of nitroblue tetrazolium reduction, and same normal cyclic AMP level as those harvested from peripheral blood . These studies describe a new disorder of granulocytes; the structural similarity to, but biochemical differences from, Chediak-Higashi disease indicate the probable heterogeneity of mechanisms for the same morphological abnormality.

Biochem J, 1983 Jun 1, 211(3), 661 - 70
Isolation and characterization of the CNBr peptides from the proteolytically derived N-terminal fragment of ovine opsin; Brett M et al.; Ovine rhodopsin may be cleaved in situ by Staphylococcus aureus V8 proteinase into two membrane-bound fragments designated V8-L (27 000 mol.wt.) and V8-S (12 000 mol.wt.) . After purification of the proteolysed complex by affinity chromatography in detergent using concanavalin A immobilized on Sepharose 4B, the two polypeptide fragments may be separated by gel-permeation chromatography on Sephadex LH-60 . Digestion of the N-terminal-derived V8-L fragment with CNBr in 70% (v/v) trifluoroacetic acid resulted in a peptide mixture that could be fractionated by procedures involving gel-permeation chromatography in organic and aqueous solvents and the use of differential solubility . The complete or partial sequences of all ten peptides are reported.

J Immunol, 1983 Jun, 130(6), 2708 - 14
The role of interleukin 1 in human B cell activation: inhibition of B cell proliferation and the generation of immunoglobulin-secreting cells by an antibody against human leukocytic pyrogen; Lipsky PE et al.; The role of factors released by monocytes (M phi) in the activation of human B lymphocytes was examined by studying the effect of an antiserum against human leukocytic pyrogen (LP) on mitogen-stimulated B cell proliferation and the generation of immunoglobulin-secreting cells (ISC) by peripheral blood mononuclear cells (PBM) . Antiserum against LP was obtained from rabbits immunized with LP-containing human M phi supernatants . The globulin fraction of this antiserum inhibited pokeweed mitogen- (PWM) stimulated B cell proliferation and the generation of ISC in a concentration-dependent manner, with 50% inhibition of responsiveness observed with 10 micrograms/ml . By contrast, PWM-induced T cell {3H}thymidine incorporation was not inhibited by concentrations of anti-LP as great as 2000 micrograms/ml . The F(ab')2 fraction of anti-LP also inhibited the generation of ISC in response to both PWM and formalinized Staphylococcus aureus, but required 50 micrograms/ml to achieve 50% inhibition . Anti-LP inhibited the generation of ISC only if present during the first 24 hr of a 6 to 7-day incubation; later addition was not inhibitory . Inhibition was more marked in cultures partially depleted of M phi than in whole PBM cultures . Whereas absorption of the anti-LP with PBM failed to remove the capacity to inhibit the generation of ISC, anti-LP-mediated inhibition of responsiveness could be reversed by the addition of crude M phi culture supernatants or a variety of highly purified interleukin 1 (IL 1) preparations, but not by T cell supernatants . These results indicate anti-LP inhibits human B cell activation by removing the requisite M phi-derived factor IL 1 and also confirm that IL 1 plays an essential role in B cell proliferation and the generation of ISC in man.

J Hosp Infect, 1983 Jun, 4(2), 149 - 57
Contamination of protective clothing and nurses' uniforms in an isolation ward; Babb JR et al.; Staphylococcus aureus was frequently isolated, usually in small numbers, from cotton gowns (12.6 per cent), plastic aprons (9.2 per cent) and nurses' uniforms (15 per cent) . Gram-negative bacilli were infrequently isolated . Contamination of the protective clothing did not increase when used over periods of up to 11 days . Fewer organisms were recovered from the front of nurses' uniforms when plastic aprons instead of gowns were worn, but gowns provided better shoulder protection . However, no differences were observed in isolations of Staph, aureus or Gram-negative bacilli from these sites.

J Hosp Infect, 1983 Jun, 4(2), 141 - 8
Isolation of anaerobic and aerobic bacteria from clean surgical wounds: an experimental and clinical study; Benediktsdottir E et al.; A pad imprint method was evaluated for quantitative sampling of anaerobic and aerobic bacteria from clean surgical wounds . The best recovery of bacteria from the pads was obtained when they were treated with a rinsing solution in a 'Stomacher' apparatus . Pads of three different fabrics were compared, but no difference was noted in their efficiency in sampling anaerobic and aerobic bacteria from clean surgical wounds . Sampling of known quantities of Staphylococcus aureus from experimental wounds in rabbits performed with the pad method gave a threefold better recovery than with a swab . The pad method was superior to the wound wash-out method when sampling wounds in hip surgery.

Infect Immun, 1983 Jun, 40(3), 1023 - 9
Production of staphylococcal enterotoxin F and pyrogenic exotoxin C by Staphylococcus aureus isolates from toxic shock syndrome-associated sources; Bonventre PF et al.; A total of 136 isolates of Staphylococcus aureus were tested for production of staphylococcal enterotoxin F (SEF) and pyrogenic exotoxin C (PEC), both of which have been identified as reliable indicators of toxic shock syndrome (TSS)-associated strains . SEF and PEC production by isolates from TSS-associated and other sources was tested independently in two laboratories, after which the two sets of data were compared . A 100% concordance between SEF and PEC production was obtained . The TSS toxin candidates were produced by 30 of 136 isolates, and in all instances SEF and PEC were made concurrently by the same strains; in no case was one toxin made and not the other . In the five groups of S . aureus tested, toxins were detected as follows: 23 of 25 (92%) acute TSS isolates, 2 of 48 (4.2%) genital non-TSS isolates, 2 of 16 (12.5%) recovered TSS isolates, 1 of 23 (4.3%) clinical nongenital isolates, and 2 of 24 (8.3%) enterotoxigenic food outbreak isolates . Comparison of purified SEF and purified PEC by immunological and biochemical criteria by immunodiffusion, isoelectric focusing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blot analysis show that the toxins are immunologically identical and strongly suggest that the two nominal TSS toxins are in fact a single protein.

JAMA, 1983 May 27, 249(20), 2803 - 7
Burn units as a source of methicillin-resistant Staphylococcus aureus infections; Boyce JM et al.; During a 3 1/2-year period (January 1978 through June 1981), 245 patients in a university hospital became colonized or infected with methicillin-resistant Staphylococcus aureus . During the first 18 months, the incidence of colonization and infection was only 0.05% . After a focal outbreak of methicillin-resistant S aureus in the burn unit, acquisition of the organism by patients on other wards increased significantly . Nonburn patients acquired the organism more often during time periods when newly admitted burn patients acquired methicillin-resistant S aureus . After the burn unit was closed, acquisition of the organism among nonburn patients on adult surgical and medical services decreased significantly . A questionnaire survey of other institutions disclosed that large outbreaks of this infection occurred more frequently in hospitals with burn units . These studies suggest that the occurrence of methicillin-resistant S aureus infections in burn units may lead to increased transmission of the organism to nonburn patients and that control of the infection in such units may reduce spread of the organism to patients without burns.

J Immunol Methods, 1983 May 27, 60(1-2), 167 - 72
Storage of protein A-conjugated sheep erythrocytes for use in plaque-forming cell (PFC) assays; Agger R et al.; Sheep red blood cells conjugated with protein A from Staphylococcus aureus were frozen in liquid nitrogen . After thawing these cells were used as indicator cells in a plaque-forming cell assay visualizing human B lymphocytes secreting IgG, IgM or IgA . The thawed erythrocytes performed as well in the assay as erythrocytes freshly conjugated with protein A . Recovery of conjugated erythrocytes after freezing and thawing averaged 66% . By the method described indicator cells for a number of PFC assays may be prepared in bulk, frozen in suitable portions and thawed when needed . This prevents possible batch-to-batch variations and saves time.

J Immunol Methods, 1983 May 27, 60(1-2), 147 - 65
Preparation and uses of immunoabsorbent monolayers in the purification of virus proteins and separation of cells on the basis of their cell surface antigens; Randall RE; A method is described for forming monolayers of Staphylococcus aureus Cowan strain A on plastic tissue culture plates (A plates) . Bacteria remain bound to such plates even under strong protein denaturing conditions . The specific binding of antibody to A plates provides a rapid method for immune precipitation on a solid matrix . Antibody can be covalently crosslinked to staphylococcus monolayers by fixation with paraformaldehyde without significant loss of specific antigen binding capacity . Furthermore antigen-antibody complexes may be efficiently disrupted with 9 M urea and non-ionic detergent and both antibody and antigen renatured by removal of urea, antibody regaining the ability to bind fresh antigen . In the presence of 9 M urea and 1% Nonidet P-40 fixed antibody remains bound to plates while antigen is released into the urea solution, providing a method for the immunological purification of proteins . Plates with such fixed antibody may be used multiple times to bind antigen . The use of this method is illustrated by the purification of 2 adenovirus structural proteins and a herpes simplex virus glycoprotein, by means of specific monoclonal antibodies to these proteins crosslinked to A plates . A method is also described to enrich for functional antibody from immune precipitates by dissociating antibody-antigen complexes with 9 M urea and 1% Nonidet P-40 and isolating the antigen free antibody on Staphylococcus aureus Cowan strain A . A plates have also been used to separate suspension cells on the basis of their cell surface antigens (e.g., lymphocyte subpopulations or cells expressing virus antigens on their surface) . Cells were either (1) reacted with specific antisera to cell surface determinants and panned over A plates, cells with antibody on their surface binding to the monolayer, or (2) panned over A plates to which specific antibody had been coupled . Separated cells may be probed directly for certain properties, such as their ability to support virus replication . The staphylococcus monolayer does not prevent the growth of tissue culture cells on the plastic surface and the selected cell population can be examined for cells giving rise to infectious centres by the subsequent addition of permissive cells.

Nucleic Acids Res, 1983 May 25, 11(10), 3065 - 76
Cloning and expression of the Staphylococcus aureus protein A gene in Escherichia coli; Duggleby CJ et al.; A gene bank of Staphylococcus aureus strain Cowan I was established using an E . coli HB101/pBR327 host-vector system . Recombinants expressing staphylococcal protein A (SPA) were detected using an IgG-binding assay . A 3.2 Kb DNA fragment directing the synthesis of SPA in E . coli was identified . SPA produced by E . coli was characterised in minicells and by Western blotting and double diffusion experiments.

J Biol Chem, 1983 May 10, 258(9), 5828 - 33
An activity from Escherichia coli membranes responsible for the modification of nitrate reductase to its precursor form; Chaudhry GR et al.; An enzymatic activity which modifies nitrate reductase has been identified in the cytoplasmic membrane of Escherichia coli . This activity changes subunit B to a form with a slightly greater electrophoretic mobility on sodium dodecyl sulfate-polyacrylamide gels (B') . The B' polypeptide produced by this modifying enzyme was compared to an apparently identical polypeptide identified in the precursor form of nitrate reductase which can be found in the cytoplasm of all strains and in the membrane of mutants defective in the insertion of nitrate reductase . These B' polypeptides were all identical with respect to mobility on gradient sodium dodecyl sulfate gels and peptides produced by limited digests using trypsin, papain, and Staphylococcus aureus V8 protease . When compared to subunit B, the proteolytic gel maps of B' polypeptides showed minor differences . From the identity of the modified B' with precursor B', the ability to convert B into B' in vitro and the in vivo nature of B' as a precursor of B, it was concluded that the modification of B to B' is a reversible process and is due to the removal of one or more small nonprotein molecules.

J Biol Chem, 1983 May 10, 258(9), 5811 - 8
The primary structure of the cytotoxin alpha-sarcin; Sacco G et al.; The primary structure of the cytotoxin alpha-sarcin was determined . Eighteen of the 19 tryptic peptides were purified; the other peptide has arginine only . The complete sequence of 17 of the peptides was determined; the sequence of the remaining peptide was determined in part . The sequence of the 39 NH2-terminal residues was obtained by automated Edman degradation . The carboxyl-terminal amino acids were identified after carboxypeptidase treatment . The assignment of the amino acids in the tryptic peptides was confirmed and their alignment established from the sequence of the secondary tryptic peptides obtained after cleavage of citraconylated alpha-sarcin, from the sequence of a 2-(2-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine peptide, from the sequence of a chymotryptic peptide, and from the sequence of a peptide obtained with Staphylococcus aureus V8 protease . alpha-Sarcin contains 150 amino acid residues; the molecular weight is 16,987 . There are disulfide bridges between cysteine residues at positions 6 and 148 and between residues 76 and 132.

Biochim Biophys Acta, 1983 May 5, 730(2), 313 - 20
Small liposomes are better than large liposomes for specific drug delivery in vitro; Machy P et al.; We have compared drug transfer into target cells in vitro from liposomes of different sizes . Liposomes of mean diameter 800 A, 2000 A or 4000 A, containing the folate analogue, methotrexate, and the fluorophore, carboxyfluorescein, were covalently coupled to Staphylococcus aureus protein A . Cells of the murine k haplotype were preincubated with an anti-H-2Kk monoclonal antibody . Excess antibody was removed and then cells were incubated with liposomes . The number of cell-bound liposomes was determined by fluorimetry . The drug effect was assayed by the methotrexate-mediated inhibition of radiolabeled deoxyuridine uptake . The drug effect was more important in the case of the 800 A vesicles than for the larger liposomes, despite the fact that the quantity of drug bound to cells was several-fold greater for large liposomes than for small ones . Since fusion is excluded by the non-proportionality of drug binding and drug effect, the predominant manner of liposome entry seems to be endocytosis . At least for these in vitro studies, the endocytosis by target cells of small liposomes seems to be more efficient than that of large liposomes.

Eur J Biochem, 1983 May 2, 132(2), 441 - 9
Identification of the lipid-binding site of phosphatidylcholine-transfer protein with phosphatidylcholine analogs containing photoactivable carbene precursors; Westerman J et al.; The lipid binding site of the phosphatidylcholine transfer protein from bovine liver has been investigated by use of phosphatidylcholine analogs which carry a diazirinophenoxy group linked to the omega-carbon of either the sn-2-{1-14C}hexanoyl (PC I) or sn-2-{1-14C}undecanoyl chain (PC II) . Photolysis of the PC I(PC II)-transfer protein complex resulted in a covalent coupling of 30-40% of the label to the protein as shown by sodium dodecyl sulfate/polyacrylamide gel electrophoresis . Upon mild alkaline treatment of the photolysed complex the protein containing covalently coupled 14C-label was separated from the noncoupled 14C-label by gel permeation chromatography . The 14C-labeled protein was degraded with protease from Staphylococcus aureus, trypsin and cyanogen bromide and specific 14C-labeled peptides were sequenced by automated Edman degradation . Major sites of coupling shown by release of radioactivity were identified as Tyr54 and the peptide segment Val171-Phe-Met-Tyr-Tyr-Phe-Asp177 . Both PC I and PC II coupled extensively to Tyr54 (90% and 50% of total labeling, respectively) . The remainder of the radioactivity was released from the peptide Val171-Asp177 with a distinct difference in in the pattern of release depending on whether PC I or PC II were used . Thus, coupling occurred preferentially to Tyr175 and Asp177 with PC I while Val171 and Met173 were labeled preferentially with PC II . This shift in coupling is compatible with an increase of 0.6 nm for the sn-2-fatty-acyl chains of PC I and II, assuming that the peptide Val171-Asp177 has adopted the strongly predicted beta-strand configuration . These data have been interpreted in terms of the localization of phosphatidylcholine in the phosphatidylcholine transfer protein.

Res Vet Sci, 1983 May, 34(3), 266 - 71
Penicillin (cloxacillin)-tolerant Staphylococcus aureus from bovine mastitis: identification and lack of correlation between tolerance in vitro and response to therapy in vivo; Craven N et al.; Twenty-four isolates of Staphylococcus aureus from naturally occurring cases of bovine mastitis were examined in vitro for resistance to cloxacillin . All strains produced penicillinase and none showed intrinsic resistance (methicillin resistance) to cloxacillin . The minimum inhibitory concentration of cloxacillin for all these strains was normal (low) but differences were found between them in their sensitivities to the bactericidal action . Three were considered to be tolerant to cloxacillin and a further five possibly so . In a mouse model of mastitis there was no difference in the response to intramammary therapy with cloxacillin between cloxacillin-sensitive or tolerant strains of S aureus and a known methicillin-resistant strain was similarly sensitive to cloxacillin therapy in vivo . It is concluded that cloxacillin tolerance may have little clinical significance in mastitis therapy.

Appl Environ Microbiol, 1983 May, 45(5), 1533 - 7
Methodology for recovery of chemically treated Staphylococcus aureus with neutralizing medium; Dey BP et al.; Recovery results of Staphylococcus aureus ATCC 6538 treated with phenolics and quaternary ammonium compounds on Dey and Engley (D/E) neutralizing medium at various time intervals were compared by the use of two commonly used media . Two recovery processes were utilized . In one, the chemically treated organisms were plated directly onto an agar medium . In the other, the aliquot was first put in broth and then was plated with agar . By either process, the numbers and the time period for recovery of organism were greater on D/E medium.

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