Lymphokine Res, 1990 Summer, 9(2), 247 - 55 Responses of pokeweed mitogen-stimulated peripheral mononuclear cells to human recombinant interleukins 3 and 4; Kato M et al.; Peripheral blood mononuclear cells (PBMC) that had been stimulated with a polyclonal B cell activator, pokeweed mitogen (PWM), were examined for mitogenic responsiveness to interleukins (IL) and colony stimulating factors (CSF) or other cytokines using recombinant preparations from E . coli or Cos cells expressing their cloned cDNAs . PWM-stimulated PBMC (PWM-blasts) were found to be responsive to IL-3 depending upon the concentrations of the agent . Mitogenic response of PWM-blasts in the reaction to IL-3 was suppressed by the addition of a specific monoclonal antibody against IL-3 but not by anti-IL-2 serum, excluding the possibility that the DNA replication induced by IL-3 is a result of the proliferative response of IL-2 responder cells to IL-2, which might be produced during the culture of PWM-blasts in the presence of IL-3 . The PWM-blasts were also responsive to IL-2 and IL-4 but not to other interleukins, namely, IL-1, IL-5 and IL-6, nor to granulocyte/macrophage (GM)-, granulocyte (G)- nor macrophage (M)-CSFs . They were also not responsive to interferon gamma . Similar responsiveness to IL-3 and IL-4 was found in the PBMC stimulated with a B-cell mitogen, Staphylococcus aureus Cowan 1 (Sac-1), but not in the PBMC stimulated with T cell mitogens, PHA or ConA . These results suggest that the human PBMC stimulated with B cell mitogens such as PWM or Sac-1 contain activated IL-3- and IL-4- responsive cells in sufficient number to detect these lymphokine activities.(ABSTRACT TRUNCATED AT 250 WORDS) Lab Delo, 1990, (2), 49 - 50 {Detection of staphylococcal protein A in broth cultures using counter electrophoresis}; Azarenok KS et al.; The diagnostic value of counter-current electrophoresis for the detection of Staphylococcus aureus protein A in the material from patients, carriers, and environmental objects is demonstrated: this method identifies 99.8% of S . aureus cultures . Microorganisms of other species and physiologic groups, not incorporating protein A, are undetectable by counter-current electrophoresis. Antimicrob Agents Chemother, 1990 Jan, 34(1), 170 - 2 Homology of mecA gene in methicillin-resistant Staphylococcus haemolyticus and Staphylococcus simulans to that of Staphylococcus aureus; Ubukata K et al.; A penicillin-binding protein of molecular weight 76,000 inducible by beta-lactams was detected in methicillin-resistant Staphylococcus haemolyticus and Staphylococcus simulans . DNA from these strains hybridized to the mecA gene from Staphylococcus aureus; however, the chromosomal HindIII fragments containing the mecA genes were 3.4 kilobases in S . haemolyticus and 4.3 kilobases in S . simulans. J Dairy Sci, 1990 Jan, 73(1), 107 - 14 Prevalence of intramammary infection and teat canal colonization in unbred and primigravid dairy heifers; Trinidad P et al.; Teat canal keratin (n = 461) and mammary gland secretions (n = 370) were collected from 31 unbred and 85 primigravid Jersey heifers from one research and three commercial dairy herds . Of 97 heifers from which secretion samples were obtained, 96.9% had intramammary infections and 29% showed clinical symptoms . Seventy-five percent of quarters were infected . Staphylococcus aureus were isolated from 36 (37.1%) heifers and 55 (14.9%) quarters . One hundred and eight (93.1%) heifers and 326 (70.7%) quarters had teat canals colonized with mastitis pathogens . Staphylococcus aureus were isolated from teat canal keratin samples from 36 (31%) heifers and 57 (12.3%) quarters . The three most common species isolated from secretion and teat canal keratin samples were Staphylococcus chromogenes, Staphylococcus hyicus, and S . aureus . Secretions from infected (n = 240) and uninfected (n = 85) quarters had SCC of 13.6 X 10(6)/ml and 5.7 X 10(6)/ml . Macrophages were the most numerous cell type in secretions of infected and uninfected quarters . Quarters with teat canal colonization, but with no intramammary infections, exhibited higher SCC in secretion (9.3 X 10(6)/ml) than quarters without both teat canal colonizations and intramammary infections (4.9 X 10(6)/ml) . Data indicated that intramammary infections and teat canal colonizations were more prevalent and SCC higher than previously realized in dairy heifers. J Immunol, 1990 Jan 1, 144(1), 60 - 5 Phenotypic and functional characteristics of human newborns' B lymphocytes; Durandy A et al.; It has been demonstrated two major facts concerning human newborns' B lymphocytes: 1) they differentiate poorly into Ig-producing cells and 2) they express CD5 and CD1c membrane proteins . We have further analyzed human newborns' B cell characteristics and found that approximately half of them express activation Ag, i.e., 4F2 and IL-2R, both associated in significant proportions with CD23 and Bac-1 . These membrane Ag were found both on CD5(+) and CD5(-) B cells . Newborns' B cells do not exhibit other activation markers because they express surface IgD, and because their size, RNA, and DNA contents do not differ from those of adults' B cells, indicating that they are in the G0/G1 cell cycle phase . Newborns' B cell proliferation can be induced by rIL-2, rIL-4, low m.w . B cell growth factor, and by Staphylococcus aureus protein A . It is presently difficult to build a hypothesis accounting for all the specific findings made on newborns' B cells . It is not known for instance whether CD5(+) and (-) B cells belong to distinct subsets as suggested by the fluorescence intensity curve obtained with an anti-CD5 antibody or to distinct stages in a unique pattern of B cell maturation during fetal and newborn life . This may indicate that partially activated B cells actually produce natural polyspecific autoantibodies of the IgM isotype found in newborns' human serum. Lett Appl Microbiol, 1990 Jan, 10(1), 27 - 9 Antibiotic sensitivity testing of bacteria by microcolony inhibition and image analysis; Hammonds SJ et al.; A rapid method for determining the sensitivity of Staphylococcus aureus to beta-lactams is described . The method involves the culture of microcolonies of the bacterium on the surface of antibiotic-containing agar medium on microscope slides . The areas of the microcolonies formed are measured by means of a microscope and image analyzer . The sensitivity of the bacterium can be estimated in 4 h by this technique and the results are comparable to those obtained by overnight incubation. J Biol Chem, 1989 Dec 25, 264(36), 21470 - 3 Two distinct Gi-proteins mediate formyl peptide receptor signal transduction in human leukemia (HL-60) cells; Gierschik P et al.; In membranes of myeloid differentiated HL-60 cells, the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine stimulates phospholipase C via a pertussis toxin-sensitive G-protein but does not inhibit adenylyl cyclase . In these membranes, the chemotactic peptide markedly stimulates the cholera toxin-dependent {32P}ADP-ribosylation of two proteins with approximate molecular masses of 40 and 41 kDa, respectively . The radiolabeled proteins comigrate on sodium dodecyl sulfate-polyacrylamide gels with the two pertussis toxin substrates present in HL-60 membranes, alpha i2 and alpha i3 . The effect of the chemotactic peptide is blocked by treatment of intact HL-60 cells with pertussis toxin . Peptide mapping studies using Staphylococcus aureus protease V8 reveal that the two radiolabeled proteins are structurally distinct . Thus, the agonist-activated formyl peptide receptor functionally interacts with two distinct pertussis toxin substrates, most likely with Gi2 and Gi3 . As the third Gi protein, Gi1, appears to be absent from both HL-60 cells and from systems that clearly reveal hormonal inhibition of adenylyl cyclase, the results strongly suggest that primary structure alone does not suffice to determine which effector mechanism is regulated by a given Gi-protein. FEBS Lett, 1989 Dec 18, 259(1), 103 - 6 Antibacterial and antimalarial properties of peptides that are cecropin-melittin hybrids; Boman HG et al.; Solid phase synthesis was used to produce 5 hybrid peptides containing sequences from the antibacterial peptide, cecropin A, and from the bee venom toxin, melittin . Four of these chimeric peptides showed good antibacterial activity against representative Gram-negative and Gram-positive bacterial species . The best hybrid, cecropin A(1-13)-melittin(1-13) was 100-fold more active than cecropin A against Staphylococcus aureus . It was also a 10-fold better antimalarial agent than cecropin B or magainin 2 . Sheep red cells were lysed by melittin at low concentrations, but not by the hybrid molecules, even at 50 times higher concentrations. J Biol Chem, 1989 Dec 15, 264(35), 21097 - 105 Limited proteolysis of recombinant human soluble interleukin-2 receptor . Identification of an interleukin-2 binding core; Miedel MC et al.; Limited proteolysis of a recombinant, soluble form of the Tac protein, a human interleukin-2 receptor (rIL-2R), was performed using trypsin, Staphylococcus aureus V8 protease and proteinase K to study the structural requirements of interleukin-2 receptor (IL-2R) for interleukin-2 (IL-2) binding . Sensitive proteolytic sites were found to be clustered in the regions of the polypeptide encoded by exons 3, 5, and 6, with a few semi-sensitive sites located within the two homologous domains encoded by exons 2 and 4 . A number of nicked and truncated rIL-2R species generated by proteolysis were assayed for IL-2 binding using recombinant IL-2 (rIL-2) affinity gel and then structurally characterized . The results demonstrated that only the species that consist of the regions encoded by exons 2 and 4, joined by five disulfide bonds, are capable of binding IL-2 and that the presence of semi-sensitive cleavage sites within the two homologous domains had no apparent effect on IL-2 binding . These results suggest that the pattern of the sensitive cleavage sites in rIL-2R is closely related to the structural requirements for IL-2 binding . Based on the experimental results, a highly symmetrical core structure of IL-2R with a total of 135 amino acid residues was identified . This is the smallest protein moiety so far known to be capable of binding IL-2. J Immunol, 1989 Dec 15, 143(12), 3988 - 93 Hyperreactivity of activated B cells to B cell growth factor in patients with systemic lupus erythematosus; Ueda Y et al.; Inasmuch as B cell function is in large part determined by lymphokine-derived accessory signals, we studied the effects of recombinant IL-2 and low-molecular-weight B cell growth factor (BCGF) on peripheral blood B cells activated with Staphylococcus aureus Cowan I to explain the B cell hyperfunction in patients with SLE . When S . aureus Cowan I-activated normal B cells were separated into Tac-antigen (Tac-Ag)+ and Tac-Ag- cells by employing a rosette technique, IL-2 induced only the Tac-Ag+ cells to proliferate, whereas both the Tac-Ag+ and Tac-Ag- cells responded to BCGF . The Tac-Ag+ and Tac-Ag- fractions of activated SLE B cells behaved like respective fractions of activated normal B cells for the pattern of response to these growth factors . It should be pointed out, however, that although the Tac-Ag+ B cells of SLE patients and those of normal controls responded to IL-2 to almost the same degree, both the Tac-Ag+ and Tac-Ag- B cells of SLE patients exhibited markedly enhanced proliferative responses to BCGF . The selectively enhanced responsiveness of a broader range of activated SLE B cells may lead to B cell hyperactivity in this disease. J Biol Chem, 1989 Dec 15, 264(35), 21205 - 10 Apolipoprotein E3-Leiden contains a seven-amino acid insertion that is a tandem repeat of residues 121-127; Wardell MR et al.; Apolipoprotein (apo) E3-Leiden is a variant of apoE that is associated with dominant expression of type III hyperlipoproteinemia and that is defective in binding to the low density lipoprotein receptor . Therefore, the structure of apoE3-Leiden was investigated . Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis apoE3-Leiden and its 22-kDa amino-terminal thrombolytic fragment migrated with a higher than normal apparent molecular weight . The structural abnormality of apoE3-Leiden was determined by sequencing its CNBr-, tryptic-, and Staphylococcus aureus V8 protease-generated peptides . In contrast to normal apoE3, which has a cysteine at residue 112, apoE3-Leiden does not contain any cysteine and has an arginine at position 112 (as does apoE4, which also completely lacks cysteine) . The basis for the molecular weight difference was determined to be a seven-amino acid insertion that is a tandem repeat of residues 121-127 of normal apoE3, i.e . Glu-Val-Gln-Ala-Met-Leu-Gly, resulting in apoE3-Leiden having 306 amino acids rather than 299 . The negatively charged glutamyl residues within the insertion compensates for the arginine substitution at residue 112; thus apoE3-Leiden focuses in the E3 position . The low density lipoprotein receptor binding activities of both intact apoE3-Leiden and its 22-kDa thrombolytic fragment were determined in an in vitro assay . Although apoE3-Leiden had only about 25% of normal binding activity, its 22-kDa thrombolytic fragment had nearly normal binding, suggesting that the carboxyl-terminal domain of apoE3-Leiden modulates the receptor binding function of its amino-terminal domain. J Biol Chem, 1989 Dec 15, 264(35), 21080 - 6 Isolation and characterization of a putative collagen receptor from Staphylococcus aureus strain Cowan 1; Switalski LM et al.; In a previous study we demonstrated that cells of Staphylococcus aureus strain Cowan bind 125I-collagen in a receptor-ligand type of interaction (Speziale, P., Raucci, G., Visai, L., Switalski, L.M., Timpl, R., and Hook, M . (1986) J . Bacteriol . 167, 77-81) . In the present communication we report on the isolation and preliminary characterization of a putative collagen receptor from a lysate of S . aureus strain Cowan . Antibodies raised against a collagen receptor positive strain inhibit the binding of 125I-collagen to bacterial cells, whereas antibodies raised against a collagen receptor negative strain were without effect . Solubilized cell surface components did not exhibit any measurable affinity for collagen-Sepharose . However, the inhibitory effect of the antibodies against bacterial cells was neutralized by the lysate from a receptor-positive but not receptor-negative strain . A collagen receptor assay was designed based on this observation and used to develop a receptor purification protocol involving anion exchange chromatography, ammonium sulfate precipitation, and gel chromatography . Using this procedure a protein with an apparent Mr of 135,000 was purified . This protein which was present on a collagen receptor-positive strain but not on a receptor-negative strain could completely neutralize the inhibitory activity of the antibodies raised against S . aureus strain Cowan . Furthermore, antibodies raised against the 135-kDa protein inhibited the binding of collagen to bacteria, and this protein is tentatively identified as a collagen receptor. Harefuah, 1989 Dec 15, 117(12), 430 - 2 {Penicillin VK absorption during fasting and after eating}; Sarel R et al.; The absorption of penicillin VK (Rafapen) was evaluated in 12 children, 3-16 years.old, with infections due to penicillin-sensitive organisms . In each, 50 mg/kg of Rafapen was given after an overnight fast, and the next day the same amount was administered after a standard hospital breakfast . Blood samples were drawn before and 30, 60 and 90 min after the drug was given . Serum penicillin levels were determined by comparing the inhibition of growth on plates of Staphylococcus aureus (Oxford strain), with that of standard penicillin G concentrations . The serum levels after the overnight fast were higher, 4.75 U/ml, as compared to 3.38 U/ml 30 min after the drug was given following breakfast . This increase was abolished at 60 and 90 min, when the serum levels were 1.96 and 1.05 U/ml, respectively . We advise the use of oral penicillin VK in infections due to penicillin-sensitive organisms, without regard to feeding schedule. Biochem J, 1989 Dec 15, 264(3), 901 - 8 Mechanisms of luteinizing-hormone exocytosis in Staphylococcus aureus-alpha-toxin-permeabilized sheep gonadotropes; van der Merwe PA et al.; We have used primary gonadotropes permeabilized with the pore-forming protein Staphylococcus aureus alpha-toxin to investigate luteinizing hormone (lutropin, LH) exocytosis . The diameter of the alpha-toxin pores (2-3 nm) allows the exchange of small molecules, whereas larger cytosolic proteins are retained . Because of the slow exchange of small molecules through the pores, we have developed a protocol which combines prolonged pre-equilibration of the permeabilized cells at 0 degrees C before stimulation with strong Ca2+ buffering . Under these conditions, increasing the free Ca2+ concentration from 0.1 microM to 10 microM {EC50 (concentration effecting half-maximal response) 2-3 microM} resulted in a 15-20-fold increase in LH exocytosis . LH exocytosis was maximal in the first 3 min and completed by 12 min . When permeabilized cells were equilibrated for prolonged periods in the absence of MgATP, Ca2(+)-stimulated LH secretion gradually declined (greater than 90% decrease by 60 min) . Addition of MgATP (5 mM) rapidly restored full Ca2(+)-stimulated LH secretion . MgATP supported Ca2(+)-stimulated LH secretion at a half-maximal concentration of 1.5 mM . UTP and adenosine 5'-{gamma-thio}triphosphate were 40 and 31% as effective as MgATP, whereas other nucleotide triphosphates were ineffective . The protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA; 50 nM) stimulated LH exocytosis at free Ca2+ concentrations as low as 1 nM and was additive with Ca2+ at higher free Ca2+ concentrations . PMA-stimulated exocytosis required MgATP at concentrations similar to those required for Ca2(+)-stimulated LH exocytosis . These results demonstrate that LH exocytosis can be triggered both by micromolar Ca2+ concentrations or, in the virtual absence of Ca2+, by PKC activation . Both mechanisms of stimulated exocytosis have an absolute requirement for millimolar ATP . Because they retain cytosolic proteins, alpha-toxin-permeabilized cells may have advantages over alternative permeabilization methods provided that conditions are used that compensate for slow diffusion through alpha-toxin pores. Biochim Biophys Acta, 1989 Dec 11, 987(1), 1 - 7 Characterization of the mitochondrial porin from Drosophila melanogaster; De Pinto V et al.; Mitochondrial porin was isolated from the fruit fly Drosophila melanogaster at different developmental stages, starting from whole mitochondria . The porin from adults' mitochondria was fully characterized . The protein had a molecular mass of 31 kDa as judged from sodium dodecylsulfate electrophoretograms . It was very resistive against digestion with V8 proteinase of Staphylococcus aureus and a larger number of fragments were only obtained after digestion with papain . Drosophila porin showed little interaction with antibodies raised against mitochondrial porins from mammalia and Neurospora crassa, but a strong reactivity with antibodies raised against yeast porin . Reconstitution experiments with planar lipid bilayer membranes showed that the protein was able to form ion-permeable pores with a single-channel conductance of 0.41 nS in 0.1 M KCl . At low transmembrane voltages Drosophila porin had the properties of a general diffusion pore with an estimated effective diameter of about 1.7 nm and a small selectivity for anions over cations . Voltages larger than 20 to 30 mV resulted in a closure of the pore . The closed states of the pore were found to be cation-selective . The addition of a synthetic polyanion to the aqueous phase on one side of the membrane resulted in an asymmetric shift of the voltage dependence and the pore became already closed at very small voltages negative at the cis-side (the side of the addition of the polyanion). Biochim Biophys Acta, 1989 Dec 8, 993(2-3), 301 - 4 Stabilization vs . degradation of Staphylococcus aureus metalloproteinase; Potempa J et al.; Purified Staphylococcus aureus metalloproteinase contains trace amounts of a serine proteinase which rapidly degrades the metalloproteinase when EDTA is present . However, no degradation occurs when Ca2+ is added or if the serine proteinase is removed by immunoaffinity chromatography . Selective chelation of Zn2+ by o-phenanthroline, which reversibly inactivates the metalloproteinase, does not result in the degradation of the apometalloproteinase, even with excess of serine proteinase . These data are interpreted as follows: EDTA chelates enzyme-bound Ca2+ and Zn2+, causing irreversible inactivation as well as a conformational change in the metal-free protein . This allows proteolysis by the contaminating serine proteinase and explains why the metalloproteinase purified from serine proteinase-deficient strains of S . aureus was previously thought to be stable to autolysis. Neurosci Lett, 1989 Dec 4, 106(3), 315 - 21 Activation of protein kinase C by 4-aminopyridine dependent on Na+ channel activity in rat hippocampal slices; Heemskerk FM et al.; The convulsant drug 4-aminopyridine (4-AP) stimulates the phosphorylation of the neuron-specific presynaptic protein B-50 in hippocampal slices . This effect could be attenuated by the protein kinase C (PKC) inhibitor staurosporine . Moreover, the endogenous phosphorylation of B-50 was found to be restricted to the 15 kDa Staphylococcus aureus protease fragment of B-50, known to contain the PKC acceptor site . The effect of 4-AP on B-50 phosphorylation was sensitive to the Na+ channel blocker tetrodotoxin . These results indicate that 4-AP stimulates PKC activity in hippocampal slices by a mechanism dependent on Na+ channel activity. J Neurochem, 1989 Dec, 53(6), 1895 - 901 Evidence for a single protein kinase C-mediated phosphorylation site in rat brain protein B-50; Coggins PJ et al.; The neuronal protein B-50 may be involved in diverse functions including neural development, axonal regeneration, neural plasticity, and synaptic transmission . The rat B-50 sequence contains 226 amino acids which include 14 Ser and 14 Thr residues, all putative sites for phosphorylation by calcium/phospholipid-dependent protein kinase C (PKC) . Phosphorylation of the protein appears to be a major factor in its biochemical and possibly its physiological activity . Therefore, we investigated rat B-50 phosphorylation and identified a single phosphorylated site at Ser41 . Phosphoamino acid analysis eliminated the 14 Thr residues because only {32P}Ser was detected in an acid hydrolysate of {32P}B-50 . Staphylococcus aureus protease peptide mapping produced a variety of radiolabelled {32P}B-50 products, none of which had the same molecular weights or HPLC retention times as several previously characterized fragments . Indirect confirmation of the results was provided by differential phosphorylation of major and minor forms of B-60 that have their N-termini at, or C-terminal to, the Ser41 residue and are the major products of specific B-50 proteolysis . Only those forms of B-60 that contained the Ser41 residue incorporated phosphate label . The results are discussed with reference to the substrate requirements for B-50 phosphorylation by PKC and the proposed structure of the B-50 calmodulin binding domain. Nihon Kyobu Shikkan Gakkai Zasshi, 1989 Dec, 27(12), 1551 - 5 {A cured case of adult respiratory distress syndrome caused by septicemia of Staphylococcus aureus}; Sakamoto K et al.; A 33-year-old woman was admitted as an emergency case because of shock condition after delivery . Physical examination revealed fever, cyanosis and coarse crackles throughout the entire chest . Laboratory data showed leukocytosis and increase of CRP value . Adult respiratory distress syndrome (ARDS) was diagnosed based on progressive severe hypoxemia and bilateral diffuse alveolar shadows on chest X-ray film . The cause of ARDS was considered to be septicemia as staphylococcus aureus was cultured from lochia . The patient was intubated, then artificially ventilated employing PEEP . Chemotherapy using three different kinds of antibiotics was started, combined with the administration of corticosteroid and a protease inhibitor, Urinastatin . The patient finally recovered completely. Ann Pediatr (Paris), 1989 Dec, 36(10), 681 - 4 {Neonatal pleuropulmonary staphylococcal infection with multiple abscesses of the liver}; Guillois B et al.; We report the case of a neonate who developed septicemia due to a methicillin-susceptible Staphylococcus aureus strain at six days of age . At eight days of age, physical evaluation revealed a tense, tender abdomen and further enlargement of the liver, and abdominal ultrasonography demonstrated multiple liver abscesses . On the following day, pleuropulmonary lesions developed and required mechanical ventilation . Recovery was achieved using parenteral vancomycin and fosfomycin for 15 days followed by oral pristinamycin for ten days . This case is unusual in that the infection was acquired by the digestive route probably as a result of lymphangitis of the breast in the mother, and multiple liver abscesses were associated with the pleuropulmonary localization . It provides further evidence of the value of ultrasonography for the diagnosis and monitoring of liver abscesses, including multiple abscesses. J Antimicrob Chemother, 1989 Dec, 24(6), 927 - 35 Intracellular activity of cefamandole and aztreonam against phagocytosed Escherichia coli and Staphylococcus aureus; Adinolfi LE et al.; The intracellular activity of cefamandole and aztreonam against phagocytosed Escherichia coli and cefamandole against phagocytosed Staphylococcus aureus was studied using a sensitive and standardized method of murine peritoneal macrophages . Cefamandole and aztreonam exerted an intracellular antibacterial activity against E . coli which was greater than their extracellular one . With concentrations of both antibiotics up to 16 x MBC a dose-dependent decrease of the initial number of intracellular E . coli which ranged from 32% to 90% was observed . However, similar antibiotic concentrations above the MBC affected the viability of extracellular E . coli by only 20% to 30% . The intracellular antibacterial activity of both antibiotics against E . coli was further enhanced by immune serum . Cefamandole at 4 x the MBC did not affect the survival of intracellular S . aureus, but killed 41% of extracellular bacteria by 1 h and 99% after 3 h . The intracellular activity of both antibiotics against E . coli was also maintained in NaF-pulsed macrophages which have an impaired oxidative metabolism . The data suggest that both cefamandole and aztreonam possess an intracellular antibacterial activity against E . coli that seems at least in part due to a positive cooperation of antibiotics with the O2-independent microbicidal system of macrophages. Antimicrob Agents Chemother, 1989 Dec, 33(12), 2155 - 6 Comparison of electronic and viability counting methods for determination of postantibiotic effect of oxacillin on Staphylococcus aureus; Nadler HL et al.; The postantibiotic effect of oxacillin on Staphylococcus aureus ATCC 6538P was determined under different test conditions by reference (viability counting) and semiautomated (electronic counting) methods . Differences in durations of the postantibiotic effect obtained with the two counting methods were not statistically significant . The semiautomated method provided a more rapid and convenient alternative to viability counting. Epidemiol Infect, 1989 Dec, 103(3), 449 - 58 Staphylococcal food poisoning from sheep milk cheese; Bone FJ et al.; Cheese made from sheep milk was implicated in food-poisoning incidents in December 1984 and January 1985 . Bacteriological examination of batches of cheese failed to reveal a viable pathogen but enterotoxin A produced by Staphylococcus aureus was present . This was the first time that enterotoxin was detected in a food produced in the UK which was associated with poisoning and from which viable Staph . aureus could not be isolated . Subsequent detailed examination of milk, yoghurt and cheese from the same producer revealed that contamination with Staph . aureus was associated with post-infection carriage as well as clinical illness in ewes on the farm . Strains producing enterotoxon . A were still intermittently present in the bulk milk used for cheese production nearly 2 years afterwards, apparently in the absence of clinical illness in the sheep . The possible effects of heat treatment are discussed . Any changes in legislation should cover all non-human mammalian milk used for human consumption. Am J Pathol, 1989 Dec, 135(6), 1129 - 37 Major histocompatibility complex protein expression on pancreas and pancreatic islet endocrine cell subsets; Lautenschlager I et al.; To determine which cells in the human and rat pancreas and islets express class I and II histocompatibility complex proteins, double indirect immunofluorescence and the Staphylococcus aureus rosette method were used . Islet preparations used permitted positive endocrine and class I or II protein identification . Class I and II proteins were expressed in pancreatic vascular endothelium and passenger cells of the mononuclear cell type . Antibodies directed to class I or beta 2 microglobulin reacted with dispersed islet B, A, and D cells, whereas class II protein antibodies were associated with only islet B and A cells . Islet B-cell class II proteins decreased after 20 days of in vitro culture . These results suggest that 1) a variety of pancreas and islet nonendocrine cells can express class I and II proteins, 2) normal pancreatic islet endocrine cells not only express class I proteins but also class II proteins, and 3) in vitro islet culture results in reduced expression of class II proteins by islet B cells. Infect Dis Clin North Am, 1989 Dec, 3(4), 901 - 13 Methicillin-resistant Staphylococcus aureus . Detection, epidemiology, and control measures; Boyce JM; MRSA are important nosocomial pathogens in many larger medical centers and are likely to find their way into hospitals that have not yet been affected . Prompt implementation of a variety of control measures shortly after the first few cases has been recognized as the approach most likely to result in the eradication of the organism . Once MRSA have become endemic, extraordinary efforts may be required to stop nosocomial transmission . For patients infected with MRSA, vancomycin is the drug of choice. Am J Dis Child, 1989 Dec, 143(12), 1424 - 7 Morbidity and mortality in children with pyogenic liver abscess; Pineiro-Carrero VM et al.; Review of our experience from 1975 to 1986 and a literature survey disclosed 109 children with pyogenic liver abscess . During this time, newer imaging techniques, especially ultrasonography and computed tomography, facilitated the prompt diagnosis of cystic lesions within the liver parenchyma . The incidence of pyogenic liver abscess at our institution (25 per 100,000 pediatric hospital admissions) was higher than previously reported . Since the majority of abscesses were located in the right lobe of the liver, patients were most effectively treated with percutaneous drainage of the abscess cavity . Staphylococcus aureus was the most common bacterial agent responsible for pyogenic liver abscess; however, anaerobic organisms were noted as a major group of pathogens and represented 27% of our patients . Furthermore, one patient was discovered to have multiple microabscesses of the liver associated with cat-scratch disease; pleomorphic gram-negative bacilli were not cultured . Among the 109 patients, the overall mortality of 15% was considerably better than that for children with PLA before 1975 . The improved survival may be related to more prompt diagnosis of pyogenic liver abscess followed by evacuation of the liver abscess and antibiotic therapy. J Surg Res, 1989 Dec, 47(6), 487 - 92 Tobramycin-adhesive in preventing and treating PTFE vascular graft infections; Shenk JS et al.; The present study was designed to determine the effectiveness of N-butyl-2-cyanoacrylate as a vehicle to deliver antibiotics locally to contaminated vascular graft sites and to grafts with established infections . Phase I--Contaminated wound model: Sixteen dogs had a 1-cm section of infrarenal aorta replaced with a PTFE graft . Prior to placement, the graft was immersed in solutions of Escherichia coli 3 X 10(8) CFU/ml and then Staphylococcus aureus 3 X 10(8) CFU/ml . After anastomosis, 1 cc of each solution was placed directly over the graft . Eleven dogs served as controls and 5 as treatment dogs . Parenteral cefonecid was given preoperatively and daily until sacrifice . Treatment animals had the anastomoses and graft sealed with a suspension of N-butyl-2-cyanoacrylate and 1.2 g tobramycin powder (antibiotic glue, ANGL) after contamination . All dogs were reoperated on the third postoperative day . Results: Eleven of 11 control dogs had positive cultures for S . aureus and 9 of 11 had positive cultures for E . coli . Seven of 11 had pseudoaneurysms, 1 exsanguinated . None of the 4 treatment dogs had positive cultures (P = 0.0002), pseudo-aneurysms (P = 0.017), or local signs of sepsis . Phase II--Infected graft model: The 10 surviving infected control dogs served as the established graft infection model . These dogs were randomized into two groups; Group 1 control (N = 5) had the graft replaced; Group 2 treatment (N = 5) had the graft replaced and ANGL treatment . Dogs were sacrificed after 2 weeks . Results: Graft cultures were positive in all 4 control dogs and negative in the 4 treatment dogs (P = 0.005) . One dog in each group was eliminated secondary to failure to obtain graft culture . The data show that ANGL can be effective in the prevention and treatment of prosthetic graft infection. Hiroshima J Med Sci, 1989 Dec, 38(4), 183 - 6 Methicillin-resistant Staphylococcus aureus in nosocomial infections in the surgical ward and operating room; Takesue Y et al.; In this study 214 strains of Staphylococcus aureus were isolated from clinical specimens on the surgical ward from 1983 to 1988 and in addition, 62 airborne strains were collected in the operating room . Highly methicillin-resistant strains of S.aureus (H-MRSA, MIC greater than 100 micrograms/ml) not detected in 1983 showed a significant increase in frequency by 1987 accounting for about 60% of MRSA (MIC greater than or equal to 12.5 micrograms/ml) . Countermeasures instituted in 1987 such as the use of disinfectant chlorhexidine alcohol significantly decreased the frequency of MRSA and H-MRSA isolates in 1988 . In our study of coagulase type, MRSA type IV strains were predominant until 1984, whereas after 1986 type II was prevalent . All airborne strains collected in the operating room were methicillin-sensitive S.aureus, with type VII currently epidemic . We therefore concluded that cross infection with MRSA took place on the surgical ward rather than in the operating room. Wei Sheng Wu Xue Bao, 1989 Dec, 29(6), 452 - 9 {The antibiotic effect of Pseudomonas jinanensis sp . nov.}; He ZZ et al.; The Pseudomonas jinanensis has been isolated from soil and screened by antibioticgram assay and spermatogonial assay . It has certain antibiotic properties . The metabolite which has been produced is certainly effective against Staphylococcus aureus and some other positive germs . The preparation of Pseudomonas jinanensis vaccine, namely PJV . It's biological activity on the growth of tumor-bearing mice and the murine spermatogenesis cell . It is considered that enhancement of nonspecific immunologic function of the host might increase the efficacy of praziquantel on schistosomiasis in mice . PJV has certainly produced effectiveness against malignant pleural effusion (effective rate was 76.5%), cancer of lung (effective rate was 58.3%), many solid kinds of cancer (effective rate was 52.1%), and the total effective rate was 64.2% . It could increase the sensitivity of cancer cells to radiation . PJV could stimulate the animal (rabbit) to develop specific antibody . PJV might belong to the class of immunomodulating agents. J Biochem (Tokyo), 1989 Dec, 106(6), 1003 - 8 The complete amino acid sequence of a subtilisin inhibitor from adzuki beans (Vigna angularis); Nozawa H et al.; The complete amino acid sequence of a major molecular form of subtilisin inhibitor from adzuki beans (Vigna angularis) was established by manual analysis using 4-N,N-dimethylaminoazobenzene-4'-isothiocyanate (DABITC) . Sequencing was performed on the peptides which were derived by digesting the inhibitor with lysyl-endopeptidase and Staphylococcus aureus V8-protease . The inhibitor consisted of 92 amino acid residues and the molecular weight was calculated to be 10,800 . A minor form of subtilisin inhibitor was found, which lacked the amino-terminal 19 residues of the major one . Comparison of amino acid sequences revealed that the adzuki bean subtilisin inhibitors were 29-68% homologous in sequence to the inhibitors of so-called "potato inhibitor I family." Nippon Seikeigeka Gakkai Zasshi, 1989 Dec, 63(12), 1545 - 51 {Protective effect of specific human immunoglobulins on haematogenous osteomyelitis by Staphylococcus aureus in mice}; Hirata H; Effects of specific human immunoglobulins extracted from pooled human sera on the protection of the production of haematogenous osteomyelitis by a strain of Staphylococcus aureus in mice were studied . No multiplication of the organisms in mouse bone marrow was observed by intravenous injection of strain SMU01 of Staphylococcus aureus when 2.20mg protein of specific immunoglobulins was administered prior to the injection of the strain . Roentgenographic examinations in mice revealed no destructive findings around the proximal and distal metaphysis of the femur at one to two weeks after the injection, indicating absence of macroscopic abscess formation in these area . These results suggest a possible therapeutic effect of specific human immunoglobulins on human staphylococcal osteomyelitis. J Okla State Med Assoc, 1989 Dec, 82(12), 622 - 5 Septic arthritis in the geriatric population; Joseph ME et al.; Septic arthritis has emerged as an important infection in geriatric patients . Between 25% and 50% of all cases of nongonococcal bacterial arthritis in nonprosthetic joints occur in patients over the age of sixty years . Thirty-three percent of nongonococcal septic arthritis occurs in the geriatric population . In this retrospective study of geriatric septic arthritis, 22 patients were reviewed . The major predisposing factor was prior history of arthritis, with the knee being the most commonly affected joint . Staphylococcus aureus was the major pathogen, with gram-negative organisms accounting for a significant number of infections . We think that empiric antibiotic coverage for gram-negative organisms should be initiated at the time of treatment only in patients with multiple predisposing factors for septic arthritis . The disease also may be more indolent than previously reported, leading to delayed diagnosis, treatment, and increased morbidity and mortality. J Antibiot (Tokyo), 1989 Dec, 42(12), 1741 - 8 New diterpenoid antibiotics, spirocardins A and B; Nakajima M et al.; New antibiotics spirocardins A and B were isolated from the culture broth of an actinomycete isolated from a soil sample collected near Lake Hibara, Fukushima Prefecture, Japan . The producing strain was classified as Nocardia sp . SANK 64282 . The antibiotics were isolated from the culture filtrate by solvent extraction and purified further by silica gel and preparative reverse phase column chromatography . They were primarily active against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus and limited species of Gram-negative bacteria such as Bacteroides fragilis and Klebsiella pneumoniae . They were also moderately active against several species of Mycoplasma . The molecular formulae of spirocardins A and B were C20H30O6 and C20H32O6, respectively . From their physico-chemical characteristics they were revealed to be diterpenoid antibiotics with closely related structures and the former was easily converted to the latter by the reduction with NaBH4. Postgrad Med J, 1989 Dec, 65(770), 941 - 2 Staphylococcus aureus meningitis from osteomyelitis of the spine; Markus HS et al.; Two cases of vertebral osteomyelitis presenting with secondary Staphylococcus aureus meningitis are described . In staphylococcal meningitis a search for a primary source should include the lower vertebral spine. J Pharm Sci, 1989 Dec, 78(12), 979 - 85 Microbioluminometry assay: determination of erythromycin activity in plasma or serum; Andreotti PE et al.; A microbioluminometry assay (MBA) was developed for the quantitative analysis of erythromycin activity in human plasma or serum . The MBA method is adapted from turbidimetric methods and utilizes an enzyme-catalyzed bioluminescence reaction to quantitate the growth of Staphylococcus aureus in liquid culture medium . Statistical analysis of data obtained in method validation studies and in more than 500 assays of standard curve and control samples demonstrates consistent reproducibility and accuracy within theoretical limits . The MBA was shown to be more sensitive than agar diffusion assays with a lower limit of sensitivity less than 20.0 ng/mL and coefficients of variation less than 10% . Cumulative results of 178 assays of spiked control plasma samples in the range of 0.14-2.18 micrograms/mL show 11.2% of individual determinations are greater than +/- 15% the expected value, and 5.6% of individual determinations are greater than +/- 20% the expected value . Bioavailability profiles obtained with MBA are consistent with reported data for erythromycin . Values for 206 subject samples analyzed by MBA and agar diffusion assays showed a high degree of correlation (r = 0.9525) between the two methods . The MBA technique provided high sample throughput because of the use of microtiter plate technologies; it is also economical since it requires less sample and reagents. J Chemother, 1989 Dec, 1(6), 377 - 83 Lincomycin resistance in methicillin-resistant Staphylococcus aureus strains of hospital origin; Spiliopoulou I et al.; A total of 170 Staphylococcus aureus strains isolated during a one-year period at the University Hospital of Patras Medical School were examined for resistance to a battery of antimicrobial agents by disk diffusion and minimum inhibitory concentration (MIC) determination . Fifty-five isolates were lincomycin- and methicillin-resistant (LMRSA) . In the group of 55 LMRSA isolates 13 were also resistant to vancomycin . All the LMRSA isolates were not typed by the international set and the experimental phages 88A and 25 at routine typing dilution (RTD), while 18 isolates were lysed by phages at 100XRTD and 1000XRTD . Reverse phage-typing and heat shock treatment of the LMRSA isolates had no effect on their typability . Plasmid profiles coupled with restriction endonuclease analysis of plasmid DNA established that the LMRSA isolates represent different strains . Membrane-protein profiles by polyacrylamide gel electrophoresis (PAGE) showed that LMRSA strains could belong to one group . This method proved useful and sensitive for characterization of LMRSA. Eur J Pediatr, 1989 Dec, 149(3), 177 - 8 Fatal infectious mononucleosis with staphylococcal pyoderma in a girl with hereditary immunological dysregulations; Inaba T et al.; We describe a 14-year-old girl with fatal infectious mononucleosis with high fever, pancytopenia, and multiple skin ulcers due to Staphylococcus aureus . Immunological studies revealed low serum IgM, low natural killer (NK) activity, and high CD4/CD8 ratio . Her father had also low NK activity and high CD4/CD8 ratio . It is suggested that she had a dominantly inherited immunodeficiency predisposing to severe Epstein-Barr virus infection. APMIS, 1989 Dec, 97(12), 1121 - 4 Erythromycin resistance in Danish Staphylococcus aureus hospital strains with emphasis on erythromycin consumption; Westh H et al.; Erythromycin resistance was analyzed in 280,415 Danish Staphylococcus aureus strains isolated from hospitalized patients and phage-typed in the period 1967 to 1987 . Multiresistant, erythromycin-resistant strains decreased during the whole period, while strains resistant only to penicillin and erythromycin increased after 1972 . This increase coincided with an increase in erythromycin consumption in Denmark from 0.4 DDD/1000 persons/day in 1978 to 1.9, in 1987 . Erythromycin consumption in nine large Danish hospitals was significantly associated with the level of erythromycin-resistant S . aureus at the hospitals. Am J Vet Res, 1989 Dec, 50(12), 2018 - 22 Further investigations into the potentiation of infection by intra-articular injection of polysulfated glycosaminoglycan and the effect of filtration and intra-articular injection of amikacin; Gustafson SB et al.; Polysulfated glycosaminoglycan (PSGAG) recently have been reported to potentiate the infectivity of Staphylococcus aureus in horses with experimentally induced septic arthritis . Four groups of 8 horses each had 1 midcarpal joint injected with approximately 33 viable colony-forming units (CFU) of S aureus plus either 1 ml of saline solution (group 1), 250 mg of PSGAG (group 2), 250 mg of PSGAG passed through a 0.6-microns filter (group 3), or 250 mg of PSGAG plus 125 mg of amikacin (group 4) . Horses that developed clinical signs consistent with sepsis were euthanatized, and samples were collected at necropsy . Horses that survived had samples obtained by use of arthroscopy at days 13 and 14 after injection . Staphylococcus aureus was isolated from 1 group-1 horse, 8 group-2 horses, and 7 of 7 group-3 horses that met protocol, but was not isolated from any group-4 horses . All 16 aforementioned horses had clinical signs, results of synovial fluid analysis, and gross pathologic and synovial membrane histopathologic findings that were consistent with septic arthritis . Polysulfated glycosaminoglycan (250 mg) increased the infectivity of 33 CFU of S aureus (P = 0.001); filtering the PSGAG had no effect . Intra-articular injection of 125 mg of amikacin immediately after inoculating the joint with 33 CFU of S aureus significantly (P = 0.001) decreased potentiation of infection by the PSGAG. Am J Vet Res, 1989 Dec, 50(12), 2014 - 7 Comparison of the effect of polysulfated glycosaminoglycan, corticosteroids, and sodium hyaluronate in the potentiation of a subinfective dose of Staphylococcus aureus in the midcarpal joint of horses; Gustafson SB et al.; Four groups of 8 horses each had 1 midcarpal joint injected with 33 colony-forming units (CFU) of viable Staphylococcus aureus plus: 1 ml of saline solution (group 1, control), 250 mg of polysulfated glycosaminoglycan (PSGAG, group 2), 100 mg of methylprednisolone acetate (group 3), or 20 mg of sodium hyaulronate (group 4) . Horses were euthanatized, and samples were obtained on the basis of clinical signs of septic arthritis that were nonresponsive to phenylbutazone administration . One group-1 horse, all 8 group-2 horses, 3 group-3 horses, and 4 group-4 horses were culture-positive for S aureus and had clinical signs, results of synovial fluid analysis, and histopathologic findings that were consistent with sepsis . The addition of 250 mg of PSGAG increased the development of sepsis significantly (P = 0.001), compared with results in control horses . Differences in the development of sepsis between horses injected with methylprednisolone acetate or sodium hyaluronate and control horses were not significant. Eur J Immunol, 1989 Dec, 19(12), 2275 - 81 Recombinant interleukin (IL) 2-induced human B cell differentiation is mediated by autocrine IL6; Xia X et al.; The molecular mechanism of the interleukin (IL) 2-induced differentiation of human B cells has been investigated . The experimental results show that Staphylococcus aureus Cowan strain I (SAC) activation alone induces IL6 secretion from B cells . When B cells were activated by SAC, there was an increased transcription of the IL6 mRNA . It reached the peak level by 6 h and rapidly decreased to an undetectable level within 24 h . The IL6 concentration in the culture supernatants reached the peak at 24-48 h and decreased slightly in the following culture periods . Since IL 2 alone could induce IgG secretion, whether exogenous IL6 was added or not, and IL2 did not increase autocrine IL6 synthesis, it appears that IL2 induces the IL6 responsiveness of SAC-activated B cells to differentiate in the later stage of the culture . The addition of polyclonal anti-IL6 antibody inhibited IgG secretion . The antibody still efficiently blocked IgG secretion up to day 5, indicating an important role of autocrine IL6 in the IL2-driven B cell differentiation . However, the saturation dose of anti-IL6 antibody inhibited 50%-70% of IgG secretion, suggesting that IL2-induced B cell differentiation appears to be mediated by other factors besides IL6. J Med Microbiol, 1989 Dec, 30(4), 245 - 52 Production of "virulence factors" by "epidemic" methicillin-resistant Staphylococcus aureus in vitro; Jordens JZ et al.; The production of virulence factors was determined quantitatively for clinical isolates of methicillin-resistant (MRSA) and methicillin-sensitive (MSSA) strains of Staphylococcus aureus from The London Hospital . The examined factors were: production of enterotoxins A, B, C and D, determined by ELISA; quantitation and differentiation of the membrane-damaging alpha, beta, gamma and delta haemolysins; and coagulase production determined by a chromogenic assay . Enterotoxin A was produced by MRSA but not by MSSA . All the strains produced haemolysins alpha, gamma and delta at similar levels, but MRSA produced significantly more coagulase than MSSA . MRSA and MSSA were compared in a phagocytosis assay but there was no difference between the phagocytosis of MRSA and MSSA by human polymorphonuclear leucocytes . These findings indicate that MRSA from The London Hospital is at least as well equipped to cause disease as other isolates of S . aureus, and probably better equipped than most hospital isolates of MSSA. Eur J Obstet Gynecol Reprod Biol, 1989 Dec, 33(3), 275 - 9 Chronic tuboovarian abscess due to Staphylococcus aureus: a case report and literature review; Meirow D et al.; A 40-year-old woman presented with a right adnexal mass . Laparotomy revealed a large tuboovarian abscess due to Staphylococcus aureus . The only significant past medical history was a Cesarean section and bilateral tubal ligation 10 years prior to admission . The tuboovarian abscess, due to this unusual organism, may have developed insidiously over a 10 year period . The patient recovered completely after excision of the abscess and antibiotic treatment. Am J Kidney Dis, 1989 Dec, 14(6), 512 - 5 Tubulointerstitial nephritis due to vancomycin; Codding CE et al.; Vancomycin was used to treat a patient with Staphylococcus aureus endocarditis . After 3 weeks of therapy, the patient developed a diffuse maculopapular rash, which resolved upon stopping the drug . Rechallenge with vancomycin several days later resulted in reappearance of the rash and rapid onset of acute anuric renal failure . Renal biopsy revealed acute granulomatous interstitial nephritis . This is the first report of biopsy-proven vancomycin-induced acute interstitial nephritis . Renal function should be monitored closely in patients receiving vancomycin therapy. J Clin Microbiol, 1989 Dec, 27(12), 2794 - 8 Rapid screening for Staphylococcus aureus infection by measuring enterotoxin B; Hosotsubo KK et al.; We developed a simple, sensitive laser nephelometric assay (LNA) to detect circulating staphylococcal enterotoxin B (SEB) . This assay yields the result within 2 h and needs no special treatment . Influence of protein A, a product generated by Staphylococcus aureus, was negligible in this assay . The levels of SEB in plasma were measured in 28 patients with and without S . aureus infection with the LNA . The levels of SEB in plasma increased significantly in patients with S . aureus infection . We also demonstrated that the levels of SEB in plasma were higher in patients with methicillin-resistant S . aureus infection than in patients with nonresistant staphylococcal infection . Our data indicate that LNA is a useful assay in the early diagnosis of methicillin-resistant S . aureus infection. J Cell Biol, 1989 Dec, 109(6 Pt 1), 2623 - 32 Identification of a human protein that interacts with nuclear localization signals; Li RH et al.; Through a series of label transfer experiments, we have identified a HeLa cell nuclear protein that interacts with nuclear localization signals (NLSs) . The protein has a molecular weight of 66,000 and an isoelectric point of approximately 6 . It associates with a synthetic peptide that contains the SV-40 T antigen NLS peptide but not with an analogous peptide in which an asparagine is substituted for an essential lysine (un-NLS peptide) . In addition to these peptides, several proteins have been tested as label donors . With the proteins, there is a correlation between nuclear localization (assayed with lysolecithin-permeabilized cells) and label transfer to the 66-kD protein . The NLS peptide (but not the un-NLS peptide) competes with the proteins in label transfer experiments, but neither wheat germ agglutinin nor ATP has an effect . These results suggest that the 66-kD protein functions as an NLS receptor in the first step of nuclear localization . In the course of this work, we have observed that the Staphylococcus aureus protein A is a strongly karyophilic protein . Its dramatic nuclear localization properties suggest that it may have multiple copies of an NLS. J Bacteriol, 1989 Dec, 171(12), 6680 - 8 Erythromycin-induced ribosome stall in the ermA leader: a barricade to 5'-to-3' nucleolytic cleavage of the ermA transcript; Sandler P et al.; The Staphylococcus aureus ermA gene, whose product confers resistance to the macrolide-lincosamide-streptogramin B family of antibiotics, is induced at the level of translation by nanomolar concentrations of erythromycin . Erythromycin also specifically stabilizes ermA transcripts, and the induced stabilization requires in-phase translation of at least one of two small leader peptides in the 5' leader region of the transcript . Erythromycin-induced mRNA stabilization was tested in three constructions in which the ermA transcript was elongated by making insertions at the ermA transcription start . Whereas mRNA downstream of the leader peptide is stabilized by erythromycin, mRNA upstream is not . In the presence of erythromycin, specific mRNA decay intermediates in both the extended ermA genes and the wild-type ermA gene were detected by both Northern blotting and S1 nuclease mapping . The 5' ends of the intermediates map to the sequences that encode each of the two ermA leader peptides, suggesting that the intermediates are produced by stalled erythromycin-bound ribosomes acting as barricades to degradation by 5'-to-3' RNases . In addition, whereas erythromycin was found previously to stabilize ermA transcripts only physically, an ermC-cat-86 hybrid transcript was stabilized both physically and functionally by erythromycin. Arch Surg, 1989 Dec, 124(12), 1456 - 9 What's in a name? Is methicillin-resistant Staphylococcus aureus just another S aureus when treated with vancomycin? McManus AT, Mason AD Jr, McManus WF, Pruitt BA Jr. Methicillin-resistant Staphylococcus aureus (MRSA) strains, principally resistant to penicillinase-resistant penicillins and aminoglycosides, are increasingly common hospital isolates . We have examined the significance of MRSA colonization and infection in 1100 consecutively admitted, seriously burned patients in whom vancomycin was used to treat all staphylococcal infections . Colonization with S aureus (SA) was identified in 658 patients, in 319 of whom MRSA colonization was identified . Two hundred fifty-three SA infections occurred in 178 patients; of these infections, 58% were pulmonic and 38% were bacteremic . Methicillin-resistant SA infections occurred in 58 of the SA-infected patients . A severity index, based on multiple-regression analysis of mortality as a function of burn size and age in the study population, was used to estimate expected mortality . We demonstrated no measurable increase in mortality attributable to MRSA in this population of burned, SA-infected patients . The results question the clinical and economic value of added control practices, such as closing of units, refusal of transfer or admission, added isolation, treatment of carriers, furlough of colonized staff, and other expensive measures that are specifically directed at prevention of MRSA infections in critical care areas. J Exp Med, 1989 Dec 1, 170(6), 2011 - 22 The staphylococcal toxic shock syndrome toxin 1 triggers B cell proliferation and differentiation via major histocompatibility complex-unrestricted cognate T/B cell interaction; Mourad W et al.; The Staphylococcus aureus exotoxin toxic shock syndrome toxin 1 (TSST-1) is a potent activator of T cells and monocytes . We have recently demonstrated that TSST-1 is a superantigen that binds monomorphic determinants on MHC class II molecules . In the present study, we have examined the effect of TSST-1 on the activation and differentiation of high density human tonsillar B cells . TSST-1 bound to tonsilar B cells with high affinity and saturation kinetics . This binding was effectively inhibited by a combination of anti-HLA-DR and anti-HLA-DQ mAbs . Treatment of purified B cells with TSST-1 failed to induce B cell proliferation or Ig production . However, in the presence of irradiated T cells, TSST-1 induced resting B cells to proliferate and differentiate into Ig secretory cells . TSST-1 mimicked nominal antigen in that its induction of B cell responses was strictly dependent on physical contact between T and B cells, and was profoundly inhibited by anti-MHC class II mAbs, anti-CD3 mAbs, and, to a lesser extent, by anti-CD18 mAbs . However, unlike nominal antigen, TSST-1-mediated T/B cell interactions were MHC unrestricted . These results suggest that TSST-1 induces T cell-dependent B cell proliferation and differentiation by virtue of its ability to mediate MHC-unrestricted cognate T/B cell interaction via the TCR/CD3 complex and MHC class II antigens. J Infect Dis, 1989 Dec, 160(6), 1061 - 3 Influence of magnesium concentration on production of exoprotein and beta-lactamase by Staphylococcus aureus and Staphylococcus hemolyticus; Kiyota H et al.; Earlier investigations demonstrated that production and secretion of toxic shock syndrome toxin-1 (TSST-1) and total exoprotein by strains of Staphylococcus aureus were maximal when magnesium ion was limiting and diminished when the concentration of magnesium increased . This investigation studied the influence of magnesium concentration on production of total exoprotein and beta-lactamase by strains of S . aureus and Staphylococcus hemolyticus, isolated from the genital tracts of women . These strains were resistant to penicillins . Each organism was incubated in chemically defined medium with various concentrations of magnesium, and total exoprotein production and beta-lactamase activity in supernatants were determined . In all strains, total exoprotein production and beta-lactamase activity per bacterial cell were markedly increased in the presence of low concentrations of magnesium . When the concentration of magnesium was elevated, production of total exoprotein and beta-lactamase was decreased . Therefore, magnesium-deficient strains of S . aureus and S . hemolyticus may secrete more exoproteins and be more resistant to beta-lactam drugs than when magnesium is not limiting. Chest, 1989 Dec, 96(6), 1273 - 9 Predictive risk factors for periannular extension of native valve endocarditis . Clinical and echocardiographic analyses; Omari B et al.; The study objective is to identify clinical, microbiologic, and/or echocardiographic risk factors present early in the course of native valve endocarditis that predict subsequent development of periannular extension of infection . A multivariate computer-generated analysis of 21 clinical-microbiologic parameters and 11 two-dimensional echocardiographic parameters in patients with native valve endocarditis was designed . These parameters were statistically compared in operated-on patients with native valve endocarditis with and without periannular extension of infection . The study took place in a 600-bed acute-care, nonreferral, municipal hospital primarily servicing an indigent patient population . Seventy-three documented episodes of native valve endocarditis occurred between the years of 1973 and 1987, including 29 operated-on patients with surgically confirmed periannular extension of infection and 44 operated-on patients without periannular extension of infection . Multivariate logistic-regression analyses of multiple clinical, microbiologic, and echocardiographic parameters which are potentially predictive of eventual periannular extension of native valve endocarditis were carried out . The only two independent parameters that significantly predicted periannular infection among patients with native valve endocarditis were (1) aortic valve involvement and (2) abuse of intravenous (IV) drugs (p less than 0.01; p less than 0.01, respectively, multivariate analysis) . The relative risk of developing periannular extension of endocarditis among patients with aortic valve involvement and/or IV drug abuse was increased by approximately 2.5-fold compared with patients without these characteristics . Factors not significantly associated with increased risk of periannular extension of native valve endocarditis included the following: prolonged febrile morbidity; Staphylococcus aureus etiology; or two-dimensional echocardiographic demonstration of vegetations, large vegetations (greater than or equal to 1 cm), multiple vegetations, or enlargement of aortic root or annulus . These data suggest that patients with native aortic valve endocarditis, particularly in the setting of IV drug abuse, should be considered for routine, serial noninvasive evaluation for the early detection of periannular extension of their infection. Clin Exp Immunol, 1989 Dec, 78(3), 366 - 71 Staphylococcus aureus-stimulated mononuclear leucocyte-conditioned medium increases the neutrophil bactericidal activity, and augments oxygen radical production and degranulation in response to the bacteria; Ferrante A et al.; Conditioned medium produced by human mononuclear leucocytes (MNL) stimulated with formalin-fixed, heat-killed Staphylococcus aureus enhanced the killing of opsonized S . aureus by human neutrophils . This enhancement was not seen when conditioned medium from non-stimulated MNL or medium cultured in the absence of both bacteria and MNL was used . There was a five-fold decrease in survival of bacteria when neutrophils were treated with conditioned medium . Conditioned medium-treated neutrophils showed increased initial rate of killing but after 20-30 min the rate of killing was similar to that of non-conditioned medium-treated leucocytes . The neutrophils treated with conditioned medium showed increased production of chemiluminescence, superoxide and lysosomal enzyme release (from both azurophilic and specific granules) in response to opsonized S . aureus . The results demonstrate that bacterial interaction with MNL leads to the release of mediators (cytokines) which potentiate the anti-bacterial function of neutrophils. J Virol, 1989 Dec, 63(12), 5509 - 13 Identification of a domain of the herpes simplex virus trans-activator Vmw65 required for protein-DNA complex formation through the use of protein A fusion proteins; Werstuck G et al.; In order to identify structural domains of the herpes simplex virus trans-activator Vmw65 required for protein-DNA complex formation, subfragments of Vmw65 were expressed in Escherichia coli as fusion polypeptides with protein A of Staphylococcus aureus, and the purified hybrids were used in a band shift assay . The results indicate that a region near the amino terminus of Vmw65 between amino acids 141 and 185 is necessary for complex formation. FEMS Microbiol Immunol, 1989 Dec, 1(8-9), 505 - 10 Detection in rabbit sera of blocking antibodies against staphylococcal fibronectin-binding protein by enzyme-linked immunosorbent assay; Luk JM et al.; Antibody titres against fibronectin-binding protein (FnBP) of Staphylococcus aureus were determined in sera from rabbits immunized with staphylococcal whole cells or purified native fibronectin receptor . An ELISA technique for detection of antibody titres blocking the binding of soluble fibronectin to immobilized FnBP was developed . A recombinant staphylococcal FnBP fused to E . coli beta-galactosidase (gal-FnBp) was used as the immobilized antigen in this test . Serum samples from two different rabbits immunized with native fibronectin receptor gave significant blocking titres, whereas the blocking titres of antisera against staphylococcal whole cells were about 4- to 5-fold lower . Using the gal-FnBP fusion protein, the sensitivity for detection of fibronectin by ELISA was also determined . The detection limit is around 5 ng . The findings are discussed with a view to developing an anti-staphylococcal adherence vaccine and quantitating fibronectin in solution. FEMS Microbiol Immunol, 1989 Dec, 1(8-9), 449 - 58 Immunoblotting of a Staphylococcus aureus DNA library as a tool for isolating potential antigenic proteins; Kothari S et al.; A Staphylococcus aureus DNA library was created in the lambda vector EMBL4 and expressed in Escherichia coli strain Y1090 . Plaques were screened using serum from a S . aureus-infected patient . A number of positive clones that expressed proteins recognised by serum antibodies were further analysed using Western blots of total lysates from cultures infected with the lambda clones . Differences were found between patient and control sera in both the specificity and titre of anti-S . aureus antibodies . Expressed staphylococcal proteins appeared to be stable and were easily distinguishable from E . coli proteins in the Western blot . This method may have applications in specifically selecting clones encoding antigens associated with infection and may be of potential use clinically. Rev Med Chil, 1989 Dec, 117(12), 1425 - 9 {Enterotoxigenic Staphylococcus aureus and thermonuclease: risk indicators of staphylococcal poisoning with milk products}; Marambio E et al.; We examined 159 samples of milk and milk products for staphylococcal count and presence of thermonuclease . Highest count for S aureus was found in cheese, followed by raw milk . Enterotoxins, mostly A and D, were found in 32% of 34 S aureus strains isolated from cheese and milk . Thermonuclease was detected in dried milk and cheese . Identification of this enzyme is a good method to evaluate risk of food poisoning from S aureus contamination. Acta Virol, 1989 Dec, 33(6), 535 - 41 Effects of phorbol myristate acetate on interleukin-2 and accompanying interferon production of human leukocytes induced by heat-inactivated Staphylococcus aureus; Rosztoczy I et al.; Interleukin-2 (IL-2) production induced by heat--inactivated Staphylococcus aureus (SAU) was enhanced by simultaneous addition of phorbol myristate acetate (PMA) . The effect was optimal at a concentration of 10 ng/ml SAU; in the presence of 10 ng/ml PMA, the amount of SAU required for maximal IL-2 production was lower . The kinetics of SAU and of SAU plus PMA-induced IL-2 production were similar . Stimulated mononuclear cells produced interferon (IFN) in addition to IL-2 . The titre of accompanying IFN was decreased in cultures stimulated with the SAU plus PMA combination . Plastic nonadherent sheep erythrocyte-positive cells were the most active in the SAU-induced IL-2 production . In contrast, the bulk of the IFN activity was produced by the nonadherent E rosette-nonforming cells . Neutralization of IFN with specific antibodies and pH 2 treatment indicated that SAU-induced IFN consisted mainly of alpha-IFN. J Dairy Sci, 1989 Dec, 72(12), 3286 - 94 Effects of recombinant granulocyte colony-stimulating factor on Staphylococcus aureus mastitis in lactating dairy cows; Nickerson SC et al.; The in vivo effects of recombinant human granulocyte colony-stimulating factor on blood and milk leukocytes in dairy cows was examined . A 2-fold increase in peripheral white blood cell counts was observed by d 5 of treatment and peaked on d 12 with values 3-fold those of controls . Counts remained elevated above pretreatment values during the treatment period, then returned to normal by d 23 of the trial . Differential white blood cell counts demonstrated that neutrophils predominated (73.8%) in treated cows versus controls (22.1%) during the treatment period . Immediately prior to experimental challenge with Staphylococcus aureus, milk SCC were 582 x 10(3) and 261 x 10(3)/ml, and percentages milk neutrophils were 64.4 and 45.3, respectively, in treated and control cows . After challenge, a 46.7% reduction in new infections was observed in quarters of treated cows compared with controls . Recombinant human granulocyte colony-stimulating factor was a granulopoietic growth and differentiation factor in the cow, and the resulting leukocytosis into the mammary gland may have been protective against experimental bacterial challenge. Scand J Immunol, 1989 Dec, 30(6), 695 - 701 Stimulation of human B lymphocyte differentiation by the neuropeptides substance P and neurokinin A; Laurenzi MA et al.; Substance P (SP) at a concentration of 10(-7) M significantly increased the number of IgG-producing cells induced by the polyclonal activator Staphylococcus aureus protein A (SpA) in 11 out of 22 cultures of enriched human blood B lymphocytes, in nine cultures SP did not significantly affect the SpA response and in three cultures IgG secretion was decreased in the presence of SP . Stimulation by SP was observed in cultures at days 6 and/or 8 . In 3 out of 4 cell cultures depleted of monocytes SP did not affect the cell response to SpA stimulation . SP antagonists inhibited the enhancing effect of SP on B-cell antibody secretion induced by SpA . SP alone did not stimulate B lymphocytes . Neurokinin A (NKA) had similar effects as SP and enhanced the IgG secretion induced by SpA in 5 out of 9 experiments, in two experiments was inactive, and in one decreased the IgG secretion . The effect of SP and NKA on B lymphocytes suggest that the neuropeptides interact with the regulation of the immune response. Biochemistry, 1989 Nov 28, 28(24), 9452 - 9 Mitochondrial NADH:ubiquinone reductase: complementary DNA sequence of the import precursor of the bovine 75-kDa subunit; Runswick MJ et al.; The 75-kDa subunit of complex I (NADH:ubiquinone oxidoreductase) from bovine heart mitochondria is its largest subunit and is a component of the iron-sulfur (IP) fragment of the enzyme . It is encoded in nuclear DNA and is imported into the organelle . Protein sequences have been determined at the N-terminus of the intact protein and on fragments generated by partial cleavage with cyanogen bromide and with Staphylococcus aureus protease V8 . Parts of these data have been used to design two mixtures of oligonucleotides 17 bases long, containing 192 and 256 different sequences, which have been synthesized and used as hybridization probes for identification of cognate cDNA clones . Two different but overlapping clones have been isolated, and the sequences of the cloned DNAs have been determined . Together they code for a precursor of the 75-kDa subunit of complex I . The mature protein is 704 amino acids in length, has a calculated molecular mass of 75,961 daltons, and contains no segments of sequence that could be folded into hydrophobic alpha-helixes of sufficient length to span the inner membrane of the mitochondrion . Its precursor has an N-terminal extension of 23 amino acids to specify its import into the mitochondrion from the cytoplasm . Seventeen cysteine residues are dispersed throughout the 75-kDa subunit; some of them are close to each other in the sequence in three separate groups and, by analogy with other iron-sulfur proteins, could be involved in iron-sulfur clusters.(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1989 Nov 25, 264(33), 20131 - 9 Sequence of the D-aspartyl/L-isoaspartyl protein methyltransferase from human erythrocytes . Common sequence motifs for protein, DNA, RNA, and small molecule S-adenosylmethionine-dependent methyltransferases; Ingrosso D et al.; A widely distributed protein methyltransferase catalyzes the transfer of a methyl group from S-adenosyl-methionine to the free carboxyl groups of D-aspartyl and/or L-isoaspartyl derivatives of L-aspartyl and L-asparaginyl residues . This enzyme has been postulated to function in the repair or the catabolism of age-damaged proteins . We present here the complete amino acid sequence of the more basic isozyme I of this enzyme from human erythrocytes . The sequence was determined by Edman degradation and mass spectral analysis of overlapping trypsin, Staphylococcus aureus V8 protease, Pseudomonas fragi endoproteinase Asp-N, cyanogen bromide, and hydroxylamine-generated fragments . The NH2-terminus is modified by acetylation and the protein contains 226 amino acids for a calculated molecular weight of 24,575 . This value is in good agreement with the molecular weight determined for the purified protein by polyacrylamide gel electrophoresis in the presence of dodecyl sulfate and by gel filtration chromatography under nondenaturing conditions . The identification of 2 different amino acid residues at both positions 22 and 119 may indicate the presence of allelic variants or of two or more closely related structural genes . Finally, comparison of this sequence with those of methyltransferases for RNA, DNA, and small molecules, as well as other S-adenosylmethionine-utilizing enzymes, shows that many of these proteins share elements of three regions of sequence similarity and may be structurally or evolutionarily related. Biochim Biophys Acta, 1989 Nov 23, 977(2), 219 - 26 Evidence that the amino-terminus of the 33 kDa extrinsic protein is required for binding to the Photosystem II complex; Eaton-Rye JJ et al.; Chymotrypsin and Staphylococcus aureus (strain V8) proteinase eliminated sixteen and eighteen amino acid residues, respectively, from the amino-terminal side of the extrinsic 33 kDa protein of the oxygen-evolving Photosystem II (PS II) complex of spinach . The carboxy-terminus of the resultant large fragments was found to be identical with that of the intact protein . Neither fragment could rebind to PS II membranes depleted of all the extrinsic proteins . Circular dichroism spectroscopy did not reveal any major conformational change within the two fragments . These results suggest that the amino-terminal region of the 33 kDa protein contains a domain essential for binding to the PS II complex. Lancet, 1989 Nov 4, 2(8671), 1071 - 3 Treatment of right-sided Staphylococcus aureus endocarditis in intravenous drug users with ciprofloxacin and rifampicin; Dworkin RJ et al.; A combination of ciprofloxacin (intravenous then oral) and oral rifampicin was tested in 14 intravenous drug users with right-sided Staphylococcus aureus endocarditis . All 10 patients who completed therapy were cured based on resolution of symptoms and negative blood cultures at 4 weeks post therapy. J Clin Microbiol, 1989 Nov, 27(11), 2574 - 81 Numerical analysis of electrophoretic protein patterns of methicillin-resistant strains of Staphylococcus aureus; Costas M et al.; A total of 50 strains of Staphylococcus aureus, including 41 methicillin-resistant S . aureus (MRSA) strains, were characterized by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins . The protein patterns contained 40-50 discrete bands and were highly reproducible . Partial patterns were used as the basis of a computer-assisted numerical analysis . The MRSA strains clustered into four phenons at the 83% similarity level; and further division of phenon 1, at the 86% similarity level, resulted in a total of six clusters . All of the MRSA isolates from an MRSA epidemic in the United Kingdom were found to cluster in phenon 1 together with 9 of the 12 MRSA isolates from eastern Australia and 3 other MRSA isolates from the United Kingdom . The remaining three eastern Australian isolates clustered separately in phenon 2 . Phenon 3 appeared to be exclusive to strains that were both susceptible and resistant to methicillin and that reacted with group V phages, and phenon 4 comprised 11 isolates, all of which were other MRSA isolates from the United Kingdom . We conclude that computer-assisted numerical analysis by high-resolution sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins provides additional criteria for the study of the epidemiology and the evolution of MRSA. J Clin Microbiol, 1989 Nov, 27(11), 2535 - 8 Use of plasmid analysis and determination of aminoglycoside-modifying enzymes to characterize isolates from an outbreak of methicillin-resistant Staphylococcus aureus; Licitra CM et al.; We compared disk susceptibility, plasmid analysis, aminoglycoside resistance patterns, and DNA hybridization for their usefulness in characterizing isolates from a hospital outbreak of methicillin-resistant Staphylococcus aureus . Fifteen isolates were susceptible (group 1) and 28 were resistant (group 2) to gentamicin . A total of 15 of 15 (100%) group 1 and 22 of 28 (79%) group 2 isolates carried a 21.5-megadalton plasmid . All group 2 isolates and none of the group 1 isolates possessed a 33-megadalton plasmid . Aminoglycoside resistance pattern determinations revealed the presence of the ANT(4')-I enzyme (aminoglycoside 4' adenyltransferase) in all group 1 isolates but was unable to demonstrate presence of this enzyme in group 2 organisms . The APH(2") + AAC(6')-II enzyme (aminoglycoside 2" phosphotransferase plus 6' acetyltransferase) was found in all of the group 2 isolates but in none of the group 1 isolates . Use of DNA hybridization revealed the presence of the ANT(4')-I enzyme in both groups (group 1, 14 of 15; group 2, 26 of 28) . In this hospital outbreak, we found good correlation between disk susceptibility, plasmid profile, aminoglycoside resistance patterns, and DNA hybridization results . It was difficult to predict the presence of the ANT(4')-I enzyme in the presence of the bifunctional {APH(2") + AAC(6')-II} enzyme by the aminoglycoside resistance pattern method because of overlap of the substrate profile. J Clin Microbiol, 1989 Nov, 27(11), 2482 - 7 Investigation by improved syringe method of effect of tampons on production in vitro of toxic shock syndrome toxin 1 by Staphylococcus aureus; Wong AC et al.; Twenty-seven types of commercial tampons from five manufacturers were tested in a sealed-syringe method to determine their effect on the growth of Staphylococcus aureus and the production of toxic shock syndrome toxin 1 . In this improvement of the syringe method, the available air is limited to that which is contained within the sealed syringe containing the tampon . The culture medium was buffered, and blood and CO2 were included in the incubation to better simulate the vaginal environment during menstruation . Variables of tampon weight, composition, air volume, and absorbency were examined for their effect on the production of toxic shock syndrome toxin 1 . Generally, with the exception of brand E, toxin production in the presence of tampons was equal to or lower than that in a sealed control syringe containing air but no tampon. J Clin Microbiol, 1989 Nov, 27(11), 2451 - 4 Phosphatidylinositol-specific phospholipase C, a possible virulence factor of Staphylococcus aureus; Marques MB et al.; Phosphatidylinositol-specific phospholipase C (PIPLC), an enzyme that can specifically release phosphatidylinositol-linked proteins from host cells, is one of the extracellular enzymes produced by Staphylococcus aureus . To investigate whether PIPLC might be a virulence factor, we assessed PIPLC production by S . aureus strains that had been isolated from healthy carriers and from infected patients with or without toxic shock syndrome . Although none of five vaginal isolates from healthy women was a PIPLC producer, only 10 of 32 selected pathogenic strains that caused significant infections or toxic shock syndrome elaborated PIPLC enzyme activity . Seven of 24 toxic-shock-associated strains, compared with 3 of 8 non-toxic-shock-associated strains, were positive for PIPLC . The majority of strains that produced PIPLC were negative for toxic shock syndrome toxin 1 (P less than 0.05); this association between PIPLC production and strains negative for toxic shock syndrome toxin 1 was even stronger among strains isolated only from patients with toxic shock syndrome (P less than 0.01) . Among all 32 pathogenic isolates, PIPLC-producing S . aureus strains were isolated from four of four patients developing adult respiratory distress syndrome and four of five patients with disseminated intravascular coagulation, suggesting a significant association between PIPLC production and adult respiratory distress syndrome and/or disseminated intravascular coagulation (P less than 0.002) . On the basis of these results, we propose that PIPLC is a virulence factor of S . aureus and is implicated in the development of adult respiratory distress syndrome and disseminated intravascular coagulation. Infect Immun, 1989 Nov, 57(11), 3512 - 9 Release of interleukin-1 beta associated with potent cytocidal action of staphylococcal alpha-toxin on human monocytes; Bhakdi S et al.; The pathogenetic relevance of Staphylococcus aureus alpha-toxin in humans has been debated because human cells have been thought to display a natural resistance toward the cytotoxic action of this cytolysin . Following our previous demonstration that human platelets represent sensitive targets for toxin attack, we have now identified monocytes as a second, highly vulnerable human cell species that succumb to attack by low doses (20 ng/ml) of alpha-toxin . The cytotoxic action of alpha-toxin is reflected in a rapid depletion of cellular ATP that is essentially complete within 30 min . The presence of human plasma proteins affords some protection of monocytes against the action of the toxin . In 10% autologous serum, ATP depletion commences at 80 to 300 ng of toxin per ml . Subcytolytic doses stimulate the release of tumor necrosis factor alpha, a process that is slightly accentuated in the presence of 50% serum . Cytocidal toxin doses unfailingly cause the release of large amounts of interleukin-1 beta from cultured cells, with levels of this monokine generally exceeding 10 ng/ml in the cell supernatants 60 min after application of toxin . Initial evidence suggests that this is due to processing of intracellular interleukin-1 rather than to de novo synthesis of the cytokine . All noted effects are abrogated in the presence of a neutralizing monoclonal antibody against alpha-toxin . Through its capacity to provoke cytokine release from monocytes and its attack on platelets, alpha-toxin may initiate cellular events that are relevant to the pathogenesis of staphylococcal infection. J Clin Endocrinol Metab, 1989 Nov, 69(5), 939 - 45 Abnormal B lymphocyte function in thyroid glands from patients with Graves' disease; Ueki Y et al.; Thyroid-infiltrating B lymphocytes from patients with Graves' disease were investigated in regard to their phenotypic profiles, cell size, cell cycle status, proliferative response to Staphylococcus aureus Cowan 1 (SAC), and spontaneous production of immunoglobulin G (IgG) and antithyroidal autoantibodies . Thyroid tissues and peripheral blood were obtained at the time of subtotal thyroidectomy of 27 Graves' patients who had been treated with thionamide drugs and iodide before operation . Two intrathyroidal mononuclear cell populations were obtained from these thyroid tissues . One cell population was isolated from the supernatants after mechanical disaggregation of the tissues and was defined as TG-1 cells . Another cell population, defined as TG-2 cells, was isolated from the supernatants of overnight cultures of the thyroid debris after enzymatic digestion . The percentages of B lymphocytes bearing activated markers and plasma cells (CD20+CD21-, IgM+IgD-, CD20+ transferrin receptor+, PCA-1+) were significantly higher in the TG-1 and TG-2 cell populations than in peripheral blood from Graves' disease patients and normal subjects . These phenotypic changes were accompanied by increased thyroid gland B lymphocyte cell size from patients with Graves' disease . The proliferative response of B lymphocytes to SAC was markedly lower in TG-1 and TG-2 cell populations than in peripheral blood cells from Graves' disease patients and normal subjects . B lymphocytes isolated from thyroid glands secreted significantly more IgG and antithyroidal autoantibodies than those from peripheral blood . Based on the findings of abnormalities in thyroid-infiltrating B lymphocytes, we suggest that activated B lymphocytes may induce the excessive production of antithyroidal autoantibodies in thyroid glands from patients with Graves' disease. Biull Eksp Biol Med, 1989 Nov, 108(11), 584 - 7 {Stimulation of immunogenesis by neurotensin, pentagastrin and thymopentin and ways of its realization}; Belokrylov GA et al.; In experiments on mice and in vitro the influence of neurotensin pentagastrin and thymopentin on the immune response, the phagocytosis of staphylococcus aureus by polymorphoneutrophil leucocytes and enzymatic activity of these cells by NBT-test were investigated . It was shown that neurotensin and thymopentin increase enzymatic and phagocytic function of polymorphoneutrophil leucocytes . Pentagastrin, as well as thymopentin stimulates the immune response, enzymatic but not phagocytic function of polymorphoneutrophil leucocytes . Immunostimulating effect of the studied peptides was realized by facility differentiation of mouse bone marrow cells into T-lymphocytes and by the interaction of the peptides with T-cells. J Ultrasound Med, 1989 Nov, 8(11), 619 - 20 Risk of Staphylococcus aureus transmission during ultrasound investigation; O'Doherty AJ et al.; An investigation of staphylococcal transmission between patients undergoing routine abdominal ultrasound procedures was carried out to assess the potential risks of cross-infection . In 13% of procedures the probe was found to be colonized with Staphylococcus aureus . Four of 19 patients were shown to have colonized the probe directly . Two patients were shown to have become colonized as a result of the procedure and some evidence is presented of possible interpatient cross-colonization . It is suggested that this is an underestimation of the potential of cross-infection and simple inexpensive infection control procedures are suggested. Virology, 1989 Nov, 173(1), 258 - 67 Characterization of an HIV-2-related virus with a smaller sized extracellular envelope glycoprotein; Rey MA et al.; A new isolate of the human immunodeficiency virus (HIV) related to the HIV-2 strain was isolated from peripheral blood lymphocytes of an Ivory Coast patient with AIDS . This isolate referred to as HIV-2 EHO could be differentiated by its envelope precursor and external glycoprotein which are 20-kDa smaller than those of HIV-2 ROD isolate . Furthermore, the apparent molecular weight of the major core protein of HIV-2 EHO is 27 kDa instead of 26 kDa as in HIV-2 ROD isolate . In addition, the product of the vpx gene which is a characteristic feature of the HIV-2 strain, is 14 kDa in HIV-2 EHO compared with 16 kDa in HIV-2 ROD . In contrast to these, the envelope precursor of HIV-2 EHO forms a transient dimer its maturation as is the case for HIV-2 ROD . In both cases the transmembrane proteins are 36 kDa and exists as homodimers of 80 kDa . Endoglycosidase H digestion experiments indicated that the 20-kDa difference between the two HIV-2 isolates is not due to a difference in the number of N-linked oligosaccharide chains per polypeptide, since deglycosylated envelope precursors of HIV-2 ROD and EHO have an apparent molecular weight of 80 and 60 kDa, respectively . Partial proteolysis of the envelope precursors from the two isolates with Staphylococcus aureus V8 protease gave a distinct pattern of polypeptides . These results suggest that the differences between the external envelope proteins of the two HIV-2 isolates are due to their amino acid composition . Accordingly, polyclonal antibodies raised against HIV-2 ROD envelope do not recognize the corresponding envelope proteins of HIV-2 EHO by immunoblotting and immunoprecipitation assays . These data illustrate that analysis of viral proteins could be useful for a rapid characterization of new viral isolates and show the heterogeneity of HIV-2 isolates in West Africa. J Clin Microbiol, 1989 Nov, 27(11), 2448 - 50 Comparative evaluation of radiometric tryptic soy broth versus radiometric tryptic soy broth with 10% sucrose for detection of bacteremia and fungemia in pediatric patients; Weinstein MP et al.; We compared BACTEC radiometric blood culture media with (8B) and without (6B) 10% sucrose for the detection of bacteremia and fungemia in pediatric patients at four university teaching hospitals that used identical methods for obtaining and processing specimens . Overall, the yields of microorganisms from 5,714 blood culture sets were no different in the two media, although a trend was noted favoring 6B for the detection of pneumococci . Speed of detection of positive results was faster in the 6B than in the 8B medium (P less than 0.05), largely due to the faster detection of Staphylococcus aureus in the 6B medium . We conclude that, overall, with pediatric patients the hypertonic 8B radiometric medium has no advantage and that it possibly has a modest disadvantage, compared with isotonic 6B radiometric medium. Mikrobiologiia, 1989 Nov-Dec, 58(6), 985 - 9 {The effect of medium acidity on the light-dependent antibacterial activity of fatty acids}; Sidorova OA et al.; The goal of this work was to study how the acidity of a medium influenced the light-dependent antibacterial activity of fatty acids towards Staphylococcus aureus . The antibacterial activity of arachidonic, linolenic, linoleic and oleic acids increased abruptly (by 1-2 orders of magnitude) under the action of visible light, it became more intensive with a rise in the number of double bonds in a fatty acid molecule, and the maximum shifted from the neutral region to an alkaline one as the aeration was intensified . In the case of saturated fatty acids (palmitic and stearic), the antibacterial activity with a maximum in the alkaline region was detected only in the light . The effect exerted by the number of double bonds in a fatty acid molecule and by the acidity of a medium on the mechanisms of fatty acid photooxidation is discussed. Biull Eksp Biol Med, 1989 Nov, 108(11), 533 - 6 {Effects of dalargin, a synthetic analog of endogenous opioids, on functional indices of isolated rat heart in experimental toxemia in vitro}; Pashutin SB; The authors studied direct effect of alpha-toxin of Staphylococcus aureus on the main functional indices of isolated perfused heart of intact Wistar rats with the working left ventricle and the ways of pharmacological correction of cardiotoxic lesion of the myocardium in vitro . It was established that alpha-toxin produced a dose-dependent effect on the functional indices of the isolated rat heart, causing in low concentrations positive inotropic effect and with more high concentrations--inhibition of cardiac activity. Bol Med Hosp Infant Mex, 1989 Nov, 46(11), 700 - 4 {Antimicrobial sensitivity profile of Staphylococcus aureus at a pediatric hospital: prevalence of resistance to methicillin}; Alpuche-Aranda C et al.; Methicillin-resistant Staphylococcus aureus (MRSA) has become a significant cause of nosocomial infections . In efforts to delineate the magnitude of this problem, we determined the prevalence of MRSA in community acquired (n = 382) and nosocomial strains (n = 207) of S . aureus isolated between Jan 1986 and March 1989 . Antimicrobial susceptibility was evaluated using an agar dilution method (Muller-Hinton agar supplemented with 4% NaCl incubated to 35 degrees C for 24 h) and MIC breakpoints were determined according to NCCLS standards . We detected (24.2%) MRSA in nosocomial strains and (5%) MRSA in community acquired strains (P less than 0.05), with a global prevalence of 11.7% . The susceptibility of community acquired S . aureus was 90% or higher for dicloxacillin, cephalothin, sulbactam/ampicillin (S/A), clindamycin, rifampicin and amikacin; 85% for cefotaxime and SMX/TMP and only 75% for erythromycin . The susceptibility pattern of the nosocomial strains was consistent with the prevalence of MRSA but the susceptibility for cephalothin, amikacin and sulbactam/ampicillin was 84.4%, 89.4% and 86.5% respectively, significantly higher than for methicillin (P less than 0.05) . Although the increased susceptibility for cephalothin and amikacin has been reported for MRSA before, the published reports using these antibiotics in the treatment of MRSA infections are controversial . The increased susceptibility of MRSA to S/A could be explained in part if the MR was mediated by "acquired MR" attributable to B-lactamase production . Our data provide a perspective on the magnitude of MRSA as a problem in a pediatric teaching hospital in Mexico . Moreover, if taken at face value, the in vitro susceptibility data point to various potential treatment options which warrant clinical evaluation. J Foot Surg, 1989 Nov-Dec, 28(6), 521 - 3 Ciprofloxacin: a study of usage in pedal infections with case reports; Notari MA et al.; Ciprofloxacin, an antibiotic of the quinolone class, has exhibited properties that may lead to frequent use in the febrile patient . Particularly important to the podiatrist is the bactericidal action of the drug on penicillinase producing Staphylococcus aureus and Pseudomonas species . Ciprofloxacin also kills many Gram-negative bacteria, and some anaerobic bacteria . Infections of this nature have been classically treated with parenteral antibiotics in a hospital setting . Ciprofloxacin has demonstrated, in clinical studies, the ability to kill these organisms in skin, soft tissues, and bone while using an oral route of administration . When considering cost-containment strategies in the patient with osteomyelitis, ciprofloxacin can cost between 50% to 80% less than parenteral therapy. J Clin Immunol, 1989 Nov, 9(6), 477 - 84 Immunological functions and phenotypes of peripheral blood lymphocytes from human T-cell leukemia virus-I carriers; Tanaka Y et al.; We studied immunological functions of peripheral blood lymphocytes (PBL) from human T-cell leukemia virus type I (HTLV-I)-seropositive healthy carriers in vitro . Proliferative responses of PBL to T-cell and B-cell mitogens such as concanavalin A (Con A), pokeweed mitogen (PWM), and Staphylococcus aureus Cowan I (SAC) were moderately impaired in HTLV-I carriers compared with normal controls . Immunoglobulin (Ig)-producing activity of PBL stimulated with B-cell mitogens were also impaired in HTLV-I carriers . However, cytotoxic T-cell activity induced by in vitro culture was not impaired but slightly increased in HTLV-I carriers . Natural killer-cell activity was only slightly decreased . By a flow cytofluorometric analysis of the cell surface phenotypes of PBL, the percentage and the mean fluorescence intensity (MFI) of CD 3-positive cells and CD 4-positive cells were significantly decreased in HTLV-I carriers . The percentage and the MFI of CD 8-positive cells was not changed . The percentage and the MFI of CD 25-positive cells were increased . These results suggest that some immunological abnormalities are already present in HTLV-I carriers and such abnormalities have some roles for the leukemogenesis from the infection of the HTLV-I into adult T-cell leukemia (ATL). Mol Gen Genet, 1989 Nov, 219(3), 480 - 5 Identification and nucleotide sequence of the delta-lysin gene, hld, adjacent to the accessory gene regulator (agr) of Staphylococcus aureus; Janzon L et al.; A Tn551 insertional mutation in the accessory gene regulator (agr) locus of the Staphylococcus aureus chromosome resulted in the decreased production of at least seven extracellular toxins and enzymes and a simultaneous increase in the production of protein A and coagulase (Recsei et al . 1986) . Adjacent to this locus we have now identified another gene, hld, transcribed into a 0.5 kb RNA which codes for the staphylococcal delta-lysin . The expression of hld was totally repressed in a strain carrying the agr insertional mutation . Hybridization with strand-specific probes and primer extension analysis revealed that hld and agr are transcribed in opposite directions, starting 188 nucleotides apart . The hld gene is mainly expressed during the post-exponential growth phase and is totally repressed during early exponential growth . Determination of hld mRNA half-life in different growth phases indicated that this regulation is at the level of transcription. Antonie Van Leeuwenhoek, 1989 Nov, 56(4), 361 - 5 Chemoattractant activity of Staphylococcus aureus serine proteinase modified human plasma alpha-1-proteinase inhibitor; Baran K et al.; S . aureus serine proteinase inactivates human alpha-1-proteinase inhibitor (alpha-1-PI) by attacking a single peptide bond between Glu354 and Ala355 giving a modified inhibitor which is a tight complex of Mr = 4,000 and 48,000 fragments . In the present paper we show that this proteolytically inactivated alpha-1-PI is a potent chemotactic factor for human neutrophiles at a nanomolar concentration, and we discuss its potential involvement in the inflammatory reaction due to S . aureus infections. J Gen Microbiol, 1989 Nov, 135 ( Pt 11), 3015 - 22 Cross-linking and O-acetylation of newly synthesized peptidoglycan in Staphylococcus aureus H; Snowden MA et al.; Staphylococcus aureus H growing exponentially was labelled with N-acetyl{14C}glucosamine, which became incorporated into the peptidoglycan . The portion of peptidoglycan not linked to teichoic acid (60-75% of the whole) was degraded with Chalaropsis muramidase to yield disaccharide-peptide monomers and dimers, trimers and oligomers formed by biosynthetic cross-linking of the monomers . The degree of O-acetylation of these fragments was also examined . Pulse-chase experiments showed that the proportion of label initially in the monomer fraction immediately after the 1 min pulse declined rapidly during a 3 min chase, while the oligomer fraction (fragments greater than trimer) gained the radioactivity proportionately . The radioactivity of the dimer and trimer fractions remained virtually unchanged . At 4 min after the commencement of labelling (i.e . approx . one-tenth of a generation time) final values had been reached . The O-acetylation of all fragments had achieved final values even at 1 min, except for the monomer fraction, which showed an increase from 40% to 60% during the first 3 min of chase . Although O-acetylation was clearly a very rapid process, no O-acetylated peptidoglycan lipid-intermediates could be detected. FEMS Microbiol Lett, 1989 Nov, 53(1-2), 143 - 7 Characterisation of a monoclonal antibody and its use in the immunoaffinity purification of penicillin-binding protein 2' of methicillin-resistant Staphylococcus aureus; Harrington CR et al.; The additional penicillin-binding protein (PBP) 2' that is important in determining intrinsic resistance in methicillin-resistant strains of Staphylococcus aureus (MRSA) has been purified by affinity chromatography using monoclonal antibodies . Monoclonal antibody 1/423.10.351 reacted in ELISA with detergent extracts of membranes from resistant organisms, but not in immunoblots with PBP 2' separated by SDS-PAGE . Immunoprecipitation experiments showed that antibody 1/423.10.351 reacted with PBP 2' in detergent extracts . The latter antibody, covalently coupled to protein A-Sepharose through the Fc region, served as an affinity matrix to purify PBP 2' . The PBP was detected in immunoblots using a second monoclonal antibody, 2/401.43 . Conjugation of this antibody with alkaline phosphatase afforded more rapid detection of PBP 2' for the immunological detection of PBP 2' both in affinity-purified fractions and in resistant strains. Arch Fr Pediatr, 1989 Nov, 46(9), 641 - 4 {Staphylococcal toxic shock in children}; Blondin G et al.; Toxic shock syndrome (TSS) is reported in 2 children . TSS occurred 10 days after an accidental injury of the fore foot in one case and 5 days following surgery for severe uretero-vesical reflux in the other . The clinical illness was defined by the case definition formulated for epidemiologic studies (CDC, 1982) . The diagnosis was confirmed by isolation of a Staphylococcus aureus strain producing TSST, at the infected site . Toxin-induced mediators such as interleukin I and Tumor Necrosis Factor have been incriminated in septic shock with multivisceral involvement . As in our 2 cases, the syndrome may be delayed or even absent. Rinsho Byori, 1989 Nov, 37(11), 1231 - 7 {Nosocomial infections and clinical microbiology}; Nakagomi O et al.; Hospital-acquired infection with strains of methicillin-resistant Staphylococcus aureus (MRSA) have considerably increased in recent years . In addition to being resistant to methicillin, these strains are resistant to practically all forms beta-lactams, aminoglycosides and many other antibiotics . There appears no cost-effective control and preventive measures for this common but also potentially life-threatening disease . Although not clearly presented, the overall cost for the treatment of patients infected with MRSA should be enormous . Can laboratory medicine (or clinical microbiology) contribute to this global medical problem? Multiple strains of MRSA circulate within a hospital and some strains are even localized within specific wards . These facts suggest yet undisclosed routes of transmission and/or foci of infection . Triumph over these versatile organisms may have to await the development of new antibiotics effective for MRSA. J Antimicrob Chemother, 1989 Nov, 24(5), 719 - 29 Penetration of macrolides into human polymorphonuclear leucocytes; Ishiguro M et al.; Five 14C-labelled macrolide antibiotics (erythromycin, josamycin, clarithromycin (TE-031), rokitamycin and roxithromycin) were studied for their transport into human polymorphonuclear leucocytes . Intracellular/extracellular concentration ratios (transport ratios) of these macrolides were quite high: erythromycin, 6.6; josamycin, 15.5; clarithromycin, 16.4; rokitamycin, 30.5; and roxithromycin, 21.9 . When polymorphonuclear leucocytes were pre-treated with formaldehyde or incubated at 4 degrees C, or at low pH, transport ratios were reduced . When extracellular macrolide was removed, intracellular macrolide concentrations became as low as 30% of the pre-wash concentrations in 5 min . KF lowered the transport ratios of josamycin and rokitamycin in particular and NaCN reduced the transport ratios of erythromycin and josamycin strikingly . Ouabain slightly lowered transport ratios of all the antibiotics tested except roxithromycin, and 2, 4-dinitrophenol decreased the transport ratio of clarithromycin markedly . The addition of various amino acids or hexose did not inhibit the transfer . Adenosine, however, inhibited the transfer of these antibiotics except erythromycin and lowered transport ratios by 83 to 92% . Puromycin reduced transport ratios of the same antibiotics by 59 to 95% . With polymorphonuclear leucocytes that had phagocytosed Legionella pneumophila serogroup 1, transport ratios of all five drugs tended to decrease . However, when Staphylococcus aureus ATCC 25923 or opsonized zymosan was phagocytosed, transport ratios for macrolides, except for roxithromycin, increased. J Antimicrob Chemother, 1989 Nov, 24(5), 637 - 45 Emergence of methicillin-resistant clones from cephamycin-resistant Staphylococcus aureus; Okonogi K et al.; Staphylococcus aureus strains specifically resistant to cephamycin antibiotics have been found among recent clinical isolates . These strains formed penicillin-binding protein (PBP) 2' and became phenotypically resistant to methicillin after induction with cefoxitin . Other cephamycin-type antibiotics also induced methicillin-resistance, whereas non-cephamycin-type cephalosporins such as cefmenoxime and ceftizoxime did not do so . The clones that constitutively synthesized PBP 2' arose from the cephamycin-resistant strains at a frequency of 10(-5) . They were indistinguishable from clinically isolated methicillin-resistant S . aureus (MRSA) . Cephamycin-resistant S . aureus may be a source for emerging MRSA. Ann Rheum Dis, 1989 Nov, 48(11), 941 - 5 B cell lymphokines in human systemic lupus erythematosus; Tan PL et al.; B lymphocytes of patients with systemic lupus erythematosus were studied to determine if they were intrinsically hyperresponsive to lymphokine mediators . Peripheral blood B cells from 25 lupus patients and 16 normal individuals matched for age and sex were cultured with recombinant lymphokines . B cells both from patients and normal subjects did not show increased {3H}thymidine uptake when cultured with interleukins 1, 2, and 4 . The addition of Staphylococcus aureus Cowan I as costimulant increased {3H}thymidine uptake by B cells of patients and normal subjects . In the absence of T cells these recombinant lymphokines did not increase in vitro IgG or IgM production by lupus or normal B cells . Other recombinant lymphokines, interleukin 3, interferon gamma, lymphotoxin, tumour necrosis factor, and colony stimulating factors for granulocytes and macrophages were tested on lymphocytes from smaller numbers of patients and controls . Most patients in this study had inactive disease and all data suggested that B cells from patients with inactive lupus were not hyperresponsive to the lymphokines tested . In addition, the use of lymphokine gene probes for interleukins 2, 3, and 4 did not show spontaneous expression of these genes in circulating lymphocytes. Exp Eye Res, 1989 Nov, 49(5), 729 - 37 Experimental bacterial endophthalmitis following extracapsular lens extraction; Records RE et al.; The rapid increase in popularity of extracapsular cataract extraction may predispose the eye to postoperative bacterial infections by introducing viable organisms through the additional instrumentation and irrigation necessary for the extracapsular technique . Lens protein released into the aqueous humor of the anterior and posterior chambers may enhance or inhibit the ability of organisms to grow in the aqueous humor . In the intact eye the lens acts as a significant protective barrier restricting the posterior extension of the infectious processes . This study was undertaken to determine if extracapsular lens extraction enhances the ability of common bacteria to infect the anterior segment of the eye and if the posterior lens capsule acts as a protective barrier denying the infectious process access to the vitreous body . Approximately 1000 colony forming units (CFU) of Staphylococcus aureus were required to produce bacterial endophthalmitis in less than one-half of normal rabbit eyes and eyes following extracapsular lens extraction . Discission of the posterior lens capsule tripled the number of eyes infected . As few as fourteen CFU could produce infections in some eyes if the posterior capsule was incised . Extracapsular lens extraction does not predispose the eye to bacterial endophthalmitis if the posterior lens capsule remains intact . Interruption of the posterior lens capsule does allow a small number of organisms to establish an intraocular infective process . Lens protein and other constituents released into the aqueous humour appear to have little effect on the growth of the test organisms. Br Vet J, 1989 Nov-Dec, 145(6), 531 - 7 Effects of dihydroheptaprenol on neutrophil functions in calves; Yoneyama O et al.; Microemulsified dihydroheptaprenol (DHP) was administered intramuscularly to clinically healthy calves at doses of 4, 6 and 8 mg/kg body weight . Marked increase of neutrophil counts was observed 0.5, 1 and 2 days after the injection at a dose of 6 mg/kg . Nitroblue tetrazolium reducing activity of neutrophils was also obviously enhanced 1, 2 and 3 days after DHP administration . Phagocytic killing of Staphylococcus aureus by neutrophils was further enhanced markedly 1, 2 and 3 days after the drug injection at 6 mg/kg. Vet Parasitol, 1989 Nov, 34(1-2), 129 - 33 Experimental production of tick pyaemia; Webster KA et al.; Lambs aged 2 weeks were inoculated with a tick-borne fever (TBF) stabilate on Day 0 and Staphylococcus aureus-contaminated ticks were applied on Day 5 . Tick pyaemia was produced experimentally for the first time using Ixodes ricinus as a mechanical vector of S . aureus . Lambs aged 18 weeks were rechallenged with a homologous strain of TBF, and S . aureus-infected ticks applied 5 days later . No significant changes were noted at post-mortem examination. J Hosp Infect, 1989 Nov, 14(4), 357 - 62 Methicillin-resistant Staphylococcus aureus: a ten-year survey in a Dublin hospital; Morgan MG et al.; We undertook a retrospective, longitudinal survey to monitor the changing incidence, phage types and patterns of antibiotic resistance of methicillin-resistant Staphylococcus aureus (MRSA) in St Laurence's Hospital (SLH), Dublin over the last 10 years . Following a peak in 1979, the incidence of MRSA has gradually decreased to around 17% of S . aureus isolates in 1986, almost identical to its incidence 10 years before . However, the spectrum of antibiotic resistance among these organisms remains broad and this seems to be at least partially related to antibiotic usage . Many strains could not be phage typed and others were typable only by the additional phage 90 . This has severely limited the usefulness of phage typing in the tracing of sources of outbreaks . Thus, there is a need for a more specific method of typing as well as a rational antibiotic policy, to successfully monitor and curtail the hospital spread of MRSA. J Hosp Infect, 1989 Nov, 14(4), 303 - 12 A laboratory-confirmed outbreak of rifampicin-methicillin resistant Staphylococcus aureus (RMRSA) in a newborn nursery; Coovadia YM et al.; The routine laboratory monitoring of methicillin-resistant strains of Staphylococcus aureus (MRSA) at a large teaching hospital led to the detection of a new, multiply-resistant strain of MRSA, which was resistant not only to penicillin, oxacillin, methicillin, cephamandole, erythromycin, tetracycline, kanamycin and gentamicin but also to rifampicin and sulphamethoxazole-trimethoprim . The rifampicin-methicillin resistant strain of S . aureus (RMRSA) was first detected in blood cultures of babies from the newborn nursery . A bacteriological investigation of the nursery revealed the source to be a paediatric medical officer who was colonised with the resistant strain, and who at the time was receiving rifampicin for pulmonary tuberculosis . The rifampicin resistance was presumably acquired du