Nutrition, 1993 Nov-Dec, 9(6), 524 - 7 Nucleoside-nucleotide mixture and its components and response to methicillin-resistant Staphylococcus aureus infection in mice; Yamamoto S et al.; The effects of a nucleoside-nucleotide mixture and its individual components on response to methicillin-resistant Staphylococcus aureus (MRSA) strain 8985N were studied in mice . BALB/c mice were fed a nucleic-acid-free 20% casein diet and intraperitoneally administered a nucleoside-nucleotide mixture, its individual components (inosine, GMP2Na, uridine, thymidine, cytidine), or saline for 30 days . On the 10th day of this treatment, the mice were inoculated intravenously with the viable MRSA organisms . By the 20th day after inoculation, mice in the saline group showed 33% survival, whereas mice in the nucleoside-nucleotide mixture group showed 75% survival . The survival rates in the inosine (62%), GMP2Na (57%), uridine (54%), cytidine (50%), and thymidine (46%) groups were not significantly different . The survival rates of the individual-component groups tended to be higher than that of the saline group, but this was not statistically different . Furthermore, there was a greater reduction in viable organism recovery in the spleen and kidney of the surviving mice in the component groups than in the saline group . However, the survival rates in the individual-component groups did not exceed that of the nucleoside-nucleotide mixture group . This study suggests that intraperitoneal administration of a nucleoside-nucleotide mixture appears to be important in producing a high survival rate after challenge with MRSA . The effect cannot be achieved with the specific components of the mixture administered individually. Rev Inst Med Trop Sao Paulo, 1993 Nov-Dec, 35(6), 551 - 5 {Staphylococcal coagglutination reaction in the identification of mycoplasma}; Timenetsky J et al.; Staphylococcal Coagglutination was used as method for a rapid identification of mycoplasmas that could be performed by non specialized laboratories . Suspensions of Staphylococcus aureus (Cowan I) sensitized with rabbit antibodies against NCTC mycoplasma strains have identified these microorganisms and the strains isolated from humans, cell cultures rats and mice in concentrated suspensions from cultures of 4.0 ml . Fourty eight strains of M.pulmonis, 6 of M . arthritidis, 8 of M.arginini, 3 of M.orale, 15 of A.laidlawii, 8 of M.hominis and 3 of M.pneumoniae were identified by staphylococcal coagglutination and confirmed by Growth Inhibition Test . Optimal parameters of coagglutination were established and the stability of the conjugates were preserved for 90 days when added with acetyl cysteine . The reaction was visualized without optical resources . The sera were previously absorbed with heterologous NCTC strains and with the pellet of the sterile broth. J Hosp Infect, 1993 Nov, 25(3), 207 - 10 Staphylococcal pneumonia in ventilated patients: a twelve-month review of cases in an intensive care unit; Inglis TJ et al.; We reviewed staphylococcal lower respiratory tract infections in our intensive care unit over a 12-month period . Staphylococcus aureus was isolated from tracheal aspirates more commonly in patients with intracranial trauma (P < 0.001), between one and six days (mean = 3) after admission to the intensive care unit . Bacteriophage typing of all S . aureus lower respiratory tract isolates from the 17 patients with head injury did not provide evidence for a common external source of infection or patient-to-patient transmission. Immunopharmacology, 1993 Nov-Dec, 26(3), 197 - 202 Cladribine (2-chloro-deoxyadenosine, CDA): an inhibitor of human B and T cell activation in vitro; Gorski A et al.; We have previously shown that the novel immunosuppressive agent cladribine (CDA) inhibits human T and B cell lymphoproliferative responses and immunoglobulin synthesis in vitro, yet appears to be particularly efficacious as an inhibitor of B cell responses . We now report the effects of CDA on the human mixed lymphocyte reaction and on expression of T and B cell activation markers . CDA produced a significant inhibition of lymphocyte proliferation in human mixed lymphocyte reactions at a concentration of 10 nM . At concentrations of 10-100 nM the drug inhibited phytohaemagglutinin-induced expression of CD25 and HLA-D (by approximately 50%), but not phorbol myristate acetate-induced expression of CD69 on purified human T cells . At a concentration of 10 nM CDA totally abolished Staphylococcus aureus Cowan-induced expression of CD25 on purified B cells . These findings confirm that CDA is a potent immunosuppressive agent with some selectivity towards B cells . The drug may have potentially wide applications in clinical immunosuppression. Antimicrob Agents Chemother, 1993 Nov, 37(11), 2278 - 84 Sequential acquisition of norfloxacin and ofloxacin resistance by methicillin-resistant and -susceptible Staphylococcus aureus; Hori S et al.; The acquisition of ofloxacin resistance by a susceptible clinical Staphylococcus aureus strain was found to be achieved in two sequential steps: the first step was accompanied by 4-fold increases in the ofloxacin MIC and 8- to 16-fold increases in the norfloxacin MIC . The second step was accompanied by further increases in both the ofloxacin and the norfloxacin MICs . A mutation of the gyrA gene resulting in an amino acid substitution was found in the second-step but not in the first-step resistant subclone . On the other hand, there was no difference in the accumulation of norfloxacin in the parent strain and the resistant subclones of each step . The rates of mutation to resistance in the steps were (1.58 to 6.81) x 10(-9) and (0.71 to 2.59) x 10(-9), respectively, and did not depend on whether the parent strain was resistant to methicillin . Some implications of these observations for clinical as well as mechanistic aspects of the prevalence of methicillin- and ofloxacin-resistant S . aureus are discussed. Biosci Biotechnol Biochem, 1993 Nov, 57(11), 1854 - 61 The complete amino acid sequence of chitinase-c from the seeds of rye (Secale cereal); Yamagami T et al.; The complete amino acid sequence of rye seed chitinase-c (RSC-c) has been analyzed . This was done by first sequencing the tryptic peptides from RCm-RSC-c and then connecting them by analyzing the peptides produced by digestions with lysylendopeptidase and Staphylococcus aureus V8 protease of RCm-RSC-c, and by chymotryptic digestion and formic acid cleavage of S . aureus V8 protease peptides . RSC-c consists of 243 amino acid residues and has a molecular mass of 26,093, and has 92% sequence homology with barley seed basic chitinase which lacks a Cys-rich domain . Cys204 is free and six cysteine residues are linked by disulfide bonds between Cys23 and Cys85, Cys97 and Cys105, and Cys223 and Cys236. Proc Natl Acad Sci U S A, 1993 Nov 1, 90(21), 9935 - 8 Lipopolysaccharide-binding protein as a major plasma protein responsible for endotoxemic shock; Gallay P et al.; Because lipopolysaccharide (LPS)-binding protein (LBP) sensitizes monocytes to LPS in vitro, it has been suggested that LBP initiates host defenses against Gram-negative bacteria . The role of LBP in vivo, and particularly in endotoxemic shock, is unknown, however . Therefore an IgG against murine LBP was prepared . It was found to neutralize binding of LPS and subsequent activation of murine macrophages in vitro . This anti-LBP protected mice against the lethal effect of LPS when given at the same time as LPS challenge, but it failed to protect mice when delayed 15 min after LPS challenge . The same preparation was also effective after challenge with lipid A but not after challenge with Staphylococcus aureus enterotoxin . The protection was correlated with a strong decrease of circulating tumor necrosis factor . These data demonstrate that in vivo LBP is a major mediator of the lethal effects of endotoxemia. J Immunol, 1993 Nov 1, 151(9), 4982 - 8 P-selectin blockade does not impair leukocyte host defense against bacterial peritonitis and soft tissue infection in rabbits; Sharar SR et al.; Adhesion molecules are responsible for PMN-endothelial cell interactions involved in both PMN-mediated endothelial injury (e.g., after ischemia-reperfusion injury) and PMN-mediated host defense against bacterial infection . Inhibition of PMN-endothelial adherence with CD18 and P-selectin mAb has been shown to ameliorate the tissue injury resulting from ischemia and reperfusion under a variety of experimental conditions . However, interference with PMN function may result in an increased risk of bacterial infection . Previous investigations suggest that CD18 blockade can lead to increased infectious risk . Little is known of the infectious risks associated with selectin blockade . We report the effects of P-selectin blockade (using mAb PB1.3) on bacteria-induced PMN emigration into the peritoneum and subcutaneous (s.c.) tissue in rabbits . Leukocyte and PMN emigration into the peritoneum 4 h after inoculation with 10 ml of 10(9) CFU/ml Escherichia coli was significant in saline-treated animals, and not different in animals pretreated with mAb PB1.3 . Similarly, the incidence and severity of abscess formation 7 days after s.c . inoculation with Staphylococcus aureus (10(7), 10(8), or 10(9) CFU) was not increased in rabbits pretreated with mAb PB1.3 compared to saline . PMN emigration to the s.c . S . aureus was also similar in both saline and mAb PB1.3-treated animals, as determined by light microscopy . We conclude that P-selectin blockade with mAb PB1.3: 1) does not interfere with acute, E . coli-induced PMN emigration into the peritoneum, 2) does not increase the incidence or severity of S . aureus abscess formation in s.c . tissue, and 3) interferes less with PMN antibacterial host defense mechanisms than inhibition of CD18-mediated PMN adherence. Biochemistry, 1993 Oct 26, 32(42), 11428 - 35 Identification and biochemical characterization of the ligand binding domain of the collagen adhesin from Staphylococcus aureus; Patti JM et al.; We have recently shown that the expression of a collagen adhesin is both necessary and sufficient to mediate the attachment of Staphylococcus aureus to cartilage, a complex collagen-containing substrate {Switalski, L . M., Patti, J . M., Butcher, W., Gristina, A . G., Speziale, P., & Hook, M . (1993) Mol . Microbiol . 7, 99-107} . We now report on the localization of the ligand binding site within the 135-kDa S . aureus collagen adhesin . Using deletion mutagenesis in combination with Western ligand blot and direct binding assays, the collagen binding domain (CBD) was localized to a 168 amino acid long segment {CBD(151-318)} within the N-terminal portion of the adhesin . Using biospecific interaction analysis, pepsin-digested bovine type II collagen was found to contain eight binding sites for CBD(151-318); two binding sites were of "high" affinity (Kd = 3 microM) and six sites were of low affinity (Kd = 30 microM) . Short truncations in the terminal flanking regions of CBD(151-318) resulted in two CBDs (180-318 and 151-297) that lacked collagen binding activity . Analysis by circular dichroism of the recombinant CBDs in the far UV revealed similar secondary structures, predominantly beta-sheet, whereas the near-UV spectra indicated dramatic changes in the degree of intermolecular packing (tertiary structure) . The deduced amino acid sequence of the ligand binding domain of the collagen adhesin is presented. Biochim Biophys Acta, 1993 Oct 20, 1182(3), 275 - 82 Pore formation by a two-component leukocidin from Staphylococcus aureus within the membrane of human polymorphonuclear leukocytes; Finck-Barbancon V et al.; The effects of the Staphylococcus aureus leukocidin (PVL), a two-component non-hemolytic toxin, were investigated on the membrane permeability of human polymorphonuclear leukocytes (PMNs) . In the absence of extracellular Ca2+, the fluorescence of ethidium bromide added to the extracellular medium increased after PVL application in a concentration-dependent manner and no variations in the free intracellular {Ca2+} of Fura2-loaded PMNs were detected . In the presence of extracellular Ca2+, the fluorescence of ethidium was not modified but the free intracellular {Ca2+} of PMNs increased after application of PVL in a concentration-dependent manner . The time lag observed before an increase in the ethidium fluorescence was longer than the time lag observed before a Fura2 fluorescence increase . Simultaneous recordings of the two probes fluorescence variations have shown the protective effect of Ca2+ and Zn2+ and the closing of the pore by 50 mM Ca2+ or 2 mM Zn2+ . Moreover, the effect of Ca2+ could be reversed by the addition of EGTA . In the presence of 1 mM extracellular Ca2+ or 0.8 mM extracellular Zn2+, the pore induced by PVL had an ionic size allowing Ca2+, Mn2+, Zn2+ and Mg2+ fluxes . The addition of antibodies against either component of PVL inhibits the permeabilization provoked by the toxin even after it was initiated . It is concluded that leukocidin from S . aureus is a pore-forming toxin which, under physiological conditions ({Ca2+} = 1 to 1.5 mM), provokes the formation of an ion-sized pore inducing an increase in the free intracellular Ca2+ which may activate PMN functions. Biochem J, 1993 Oct 15, 295 ( Pt 2), 437 - 45 Biochemical characterization of hamster oviductin as a sulphated zona pellucida-binding glycoprotein; Malette B et al.; Oviductins are a family of glycoproteins, synthesized and released by oviductal secretory cells, which bind to the zona pellucida of the oocyte after ovulation . Hamster oviductin migrates as diffuse species of 160-350 kDa during SDS/PAGE under reducing as well as non-reducing conditions . In this report, we describe the one-step purification of hamster oviductin using either immuno- or lectin-affinity chromatography . Probing with specific lectins showed that the glycoprotein contains terminal alpha-D-GalNAc, and either terminal alpha-D-NeuAc or non-terminal beta-D-(GlcNAc)2 residues, but fails to react with concanavalin A and Ulex Europeus A-1 lectins which are specific for branched alpha-D-mannose and alpha-L-fucose moieties respectively . Intraovarian oocytes do not contain this glycoprotein and we demonstrate here that the immunoaffinity-purified oviductin readily binds to their zonae pellucidae in vitro, thus mimicking the in vivo phenomenon . Two major immunologically related forms of hamster oviductin (named alpha and beta) were characterized using one- and two-dimensional gel electrophoresis . The alpha-form (160-210 kDa) has an acidic pI of 3.5-4.5 and the beta-form (approx . 210-350 kDa) is localized at the cathodic site in the isoelectric focusing dimension; in between these two major forms lies a smear of minor-charge isomers . Peptide mapping of both major forms with papain and Staphylococcus aureus V8 protease yielded fragments of identical size . Moreover, the two forms share the same N-terminal sequence which display no significant homology with other reported proteins . Treatment with trifluoromethanesulphonic acid showed that a protein with the size and pI of the alpha-form can be generated from the beta-form . Both the alpha- and beta-forms are sulphated on O-linked oligosaccharide side chains but are not phosphorylated . Collectively, these results suggest that the hamster oviductin polymorphism observed in two-dimensional PAGE is a consequence of different glycosylation patterns and not the polypeptide chain itself . Hamster oviductin is mostly O-glycosylated and contains a few N-linked oligosaccharide side chains (approx . 10 kDa) . We propose that hamster oviductin is a mucin-type glycoprotein which might act as a protective secretion influencing the first steps of the reproductive process necessary for the normal triggering of fertilization and early embryonic development. Biochem Biophys Res Commun, 1993 Oct 15, 196(1), 473 - 9 Purification and characterization of a catalytically active human liver UDP-glucuronosyltransferase expressed as a fusion protein in E . coli; Pillot T et al.; The purification and the characterization of functional human liver UDP-glucuronosyltransferase 2B4 produced as a Staphylococcus aureus protein A fusion protein in E . coli are described . The purified fusion protein was able to catalyze the glucuronidation of hyodeoxycholic acid, the major substrate described for this isoform to date . The effects of the amount and the nature of the phospholipids upon reconstitution into phospholipid micelles were investigated . Apparent determined Km values for hyodeoxycholic acid and UDP-glucuronic acid were 0.55 and 0.43 mM, respectively . Moreover, photoaffinity labelling of the fusion protein with a photoactivatable analog of UDP-glucuronic acid strongly suggested that this recombinant protein exhibited similar binding properties as the microsomal protein, which emphasizes its use for further structural analyses. Nucleic Acids Res, 1993 Oct 11, 21(20), 4830 - 5 The mouse antiphosphotyrosine immunoreactive kinase, TIK, is indistinguishable from the double-stranded RNA-dependent, interferon-induced protein kinase, PKR; Baier LJ et al.; The mouse TIK protein, a serine/threonine kinase, was originally isolated from a murine pre-B cell expression library by its ability to bind anti-phosphotyrosine antibodies (Icely et al., J . Biol . Chem . 266, 16073-16077, 1991) . The 67 kDa protein was found to have an associated autophosphorylation activity when incubated with ATP . Our results show that TIK is actually the mouse interferon-induced, dsRNA-dependent protein kinase, PKR . We demonstrate that the TIK message is interferon-inducible in mouse L-cells and in vitro transcription and translation of the TIK cDNA produces a protein that is capable of binding double-stranded RNA . The in vitro synthesized TIK protein migrated as a 65 kDa protein on SDS-PAGE when incubated with ATP, but migrated as a 60 kDa protein when incubated with an inhibitor of PKR, 2-aminopurine . We further show that proteolytic digestion of TIK with Staphylococcus aureus V8 protease results in a cleavage pattern identical to that obtained by V8 digestion of authentic PKR . Antiserum to TIK specifically recognized PKR . Cloned TIK had inhibitory activity for replication of EMCV but not VSV . From these observations we conclude that TIK kinase is the mouse interferon-induced, double-stranded RNA-dependent kinase, PKR. Med Clin (Barc), 1993 Oct 2, 101(10), 379 - 82 {Mitral endocarditis caused by Staphylococcus aureus resistant to methicillin, aminoglucosides and rifampicin: description of 2 cases with fatal course}; Serrano R et al.; Two patients with infectious endocarditis (IE) by Staphylococcus aureus resistant to methicillin, aminoglucosides and rifampicin (SARMAR) acquired in hospital during the course of an epidemic outbreak of this microorganism in the Hospital Clinic i Provincial of Barcelona . Both patients had undergone surgery of the lower limbs . The entrance of the microorganism was the infection of the surgical wound, with bacteriemia, followed by mitral IE after a short time interval (20 days) . Despite adequate treatment with vancomycin both patients died . The culture of mitral vegetation was positive for SARMAR in one . Analysis of the chromosomic DNA of all the isolations from the patients was identical and coincided with that of the SARMAR strains isolated in the epidemic outbreak of the hospital . The current situation of IE by SARMAR is reviewed and the therapeutic implications commented upon suggesting that treatment of this entity should simultaneously include the administration of vancomycin and phosphomycin or cotrimoxazole, with surgery being considered if infection persists. J Med Microbiol, 1993 Oct, 39(4), 305 - 9 Lectin typing of methicillin-resistant Staphylococcus aureus from Singapore, England and Wales, and Denmark; Jarlov JO et al.; Methicillin-resistant Staphylococcus aureus (MRSA) isolates from a possible outbreak in Singapore (84) were examined together with all MRSA isolated in Denmark in 1986-1990 (58) and 14 distinct epidemic and 10 distinct single hospital strains from England and Wales . All 84 Singapore isolates were phage typed routinely and 52 isolates were further analysed together with the Danish isolates with an additional set of experimental phages and by lectin typing . The British strains, previously phage typed in the same way, were lectin typed . The following lectins were used: wheat-term agglutinin, soy-bean agglutinin, tomato lectin and Concanavalin-A . Routine phage typing of the Danish isolates showed that 41 isolates belonged to 19 different types; 17 isolates were non-typable (NT) . Addition of experimental phage typing and lectin typing enhanced discrimination to 47 types . The 24 British strains could be divided into eight {corrected} "lectin types" . Sixty-one of the isolates from Singapore were non-typable by phage typing; the remaining 23 strains belonged to five types . Further examination of 52 isolates with the experimental set of phages and by lectin typing gave 14 closely related types; 48% of these isolates belonged to only two types. Infect Immun, 1993 Oct, 61(10), 4254 - 62 Characterization of novel type C staphylococcal enterotoxins: biological and evolutionary implications; Marr JC et al.; The type C staphylococcal enterotoxins (SEC) are a group of highly conserved proteins with significant immunological cross-reactivity . Although three antigenically distinct SEC subtypes (SEC1, SEC2, and SEC3) have been reported in the literature, we observed that the isoelectric points of SEC from several Staphylococcus aureus isolates are different from those of any of these three subtypes . This observation led us to propose that additional SEC molecular variants exist . For assessment of this possibility, the sec genes from representative human, animal, and food isolates were cloned and sequenced . The toxins encoded by the 18 isolates used in this study included five unique SEC proteins in addition to SEC1, SEC2, and SEC3 . Six of the SEC proteins (including SEC1, SEC2, and SEC3) were produced by human and food isolates . Analysis of seven bovine and ovine isolates showed that isolates from each animal species produced a unique host-specific SEC . All of the SEC caused lymphocyte proliferation, although some of the toxins differed in their ability to stimulate cells from several animal species . An explanation for these results, which is supported by our phenotypic analysis of Sec+ staphylococcal isolates, is that toxin heterogeneity is due to selection for modified SEC sequences that facilitate the survival of S . aureus isolates in their respective hosts. Exp Hematol, 1993 Oct, 21(11), 1467 - 72 The effects of treatment with PIXY321 (GM-CSF/IL-3 fusion protein) on human polymorphonuclear leukocyte function; Buescher ES et al.; During a phase I trial of the genetically engineered hematopoietic growth factor PIXY321 (granulocyte-macrophage colony-stimulating factor/interleukin-3 {IL-3} fusion protein), we examined the effects of PIXY321 treatment on human polymorphonuclear leukocyte (PMN) locomotive, respiratory burst, and phagocytic responses . PIXY321 treatment was associated with transient suppression of both unstimulated locomotion and chemotaxis responses to multiple stimuli, as well as significant transient enhancement of formyl peptide-stimulated H2O2 production . No effects on opsonic phagocytosis of Staphylococcus aureus were observed . In vitro exposure of control PMN to PIXY321 resulted in suppression of unstimulated locomotion/chemotaxis and enhancement of formyl peptide-stimulated H2O2 production but had no effects on phagocytosis . When patient cells were exposed in vitro to PIXY321 during treatment, suppression of chemotaxis and enhancement of H2O2 production were observed before PIXY321 treatment, but these effects diminished during treatment . The in vivo and in vitro exposure effects of PIXY321 treatment on PMN function are similar to those of the parent molecule, granulocyte-macrophage colony-stimulating factor (GM-CSF). Obstet Gynecol, 1993 Oct, 82(4 Pt 2 Suppl), 660 - 2 Toxic shock syndrome associated with vulvar necrotizing fasciitis; Farley DE et al.; BACKGROUND: Fifty percent of toxic shock syndrome is associated with nonmenstrual etiologies such as postoperative wound infection . CASE: A 44-year-old woman developed necrotizing vulvar fasciitis that was successfully treated with surgical debridement and broad-spectrum antibiotics . However, after improving for 3 days postoperatively, she developed fever, a generalized maculopapular rash, and renal and liver abnormalities . As her condition worsened, she developed hypotension and respiratory distress . After 5 days in the intensive care unit, she gradually improved . Her wound culture from admission grew multiple organisms, including Staphylococcus aureus that produced toxic shock syndrome toxin-1 . CONCLUSION: Toxic shock may occur in varied gynecologic settings, including pelvic and perineal infection . Successful management requires a prompt and aggressive response to multi-organ system failure. J Infect Dis, 1993 Oct, 168(4), 961 - 8 Exposure to ethanol up-regulates the expression of Mycobacterium avium complex proteins associated with bacterial virulence; Bermudez LE et al.; Mycobacterium avium complex (MAC) organisms are the most common bacterial cause of disseminated infection in patients with AIDS . MAC, facultative intracellular bacteria, invade and multiply within macrophages . Treatment of MAC-infected macrophages with ethanol (10-100 micrograms/dL) is associated with increased intracellular multiplication of MAC . To investigate whether this enhanced growth is due to a stress-related response induced by nonlethal concentrations of ethanol, strain 101 (serovar 1) was exposed to ethanol, and the regulation of the expression of proteins was examined . Exposure of MAC to ethanol (range, 10-100 micrograms/dL) was associated with up-regulation of the expression of a number of bacterial proteins, some of which (65 and 33 kDa) interfered with macrophage functions, such as production of superoxide anion and killing of Staphylococcus aureus . Thus, exposure of MAC to small concentrations of ethanol may induce a stress-related response with consequent increase in the synthesis of proteins possibly associated with its ability to survive within macrophages. J Infect Dis, 1993 Oct, 168(4), 893 - 6 The effect of zinc on microbial growth and bacterial killing by cefazolin in a Staphylococcus aureus abscess milieu; Bamberger DM et al.; Microbial growth and antimicrobial bacterial killing are both diminished in abscesses . It was postulated that zinc depletion in abscesses, perhaps secondary to a neutrophil protein resembling calprotectin, may be partly responsible for these effects . In a rabbit tissue-cage abscess model, pooled abscess supernatant concentration of zinc was < 1.53 microM . The addition of 41.7 microM zinc had no effect on Staphylococcus aureus growth or the bacterial killing effect of cefazolin in serum . In abscess fluid supernatants, bacterial growth without antibiotic and bacterial killing by cefazolin were both enhanced by the addition of zinc . Fractionation of the abscess fluid with ultrafiltration membranes showed that these effects could be reproduced with the fraction between 30 and 50 kDa . These findings suggest that a protein in abscess fluid supernatants that resembles the neutrophil protein calprotectin may, through its zinc binding effects, inhibit microbial growth within an abscess but also inhibit the activity of bactericidal antibiotics. J Infect Dis, 1993 Oct, 168(4), 1030 - 3 Passive local immunotherapy of experimental staphylococcal pneumonia with human intravenous immunoglobulin; Ramisse F et al.; Staphylococcus aureus remains a life-threatening agent of nosocomial pneumonia in immunocompromised patients . The increasing incidence of strains exhibiting wide-spectrum resistance to antibiotics, such as methicillin-resistant S . aureus (MRSA), requires new therapeutic strategies . There is renewed interest in passive immunization with human plasma-derived immunoglobulins (IVIG) as antiinfective agents . The efficacy of IVIG was tested in an experimental model of staphylococcal pneumonia, using both an MRSA clinical isolate and reference strain Cowan III, in mice immunosuppressed with cyclophosphamide . Efficient antistaphylococcal activities were obtained with IVIG administered intravenously or intranasally . IVIG saturated with protein A or its F(ab')2 fragments were as efficient as intact IVIG, suggesting that protection did not require opsonization through IgG Fc-phagocyte Fc gamma-receptor interactions . Thus, topical administration of IVIG may replace a local antibody response to S . aureus in an immunocompromised host and may be useful in prophylaxis and treatment of nosocomial S . aureus pneumonia. J Immunol, 1993 Oct 1, 151(7), 3478 - 88 Regulation of C gamma subclass germ-line transcripts in human peripheral blood B cells; Kitani A et al.; In the present study we investigated the early steps of human IgG subclass differentiation by defining the conditions necessary for IgG subclass-specific production of germ-line transcripts in human peripheral blood B cells . Constant region of gamma-globulin (C gamma) subclass germ-line transcripts were measured using newly developed reverse transcription and polymerase chain reaction (RT-PCR) assays that were shown to be C gamma subclass-specific germ-line transcripts by size of the amplification products obtained before and after digestion with appropriate endonucleases (NarI and NcoI) . In initial studies we found that C gamma 1 and C gamma 2 germ-line transcripts were constitutively expressed in total peripheral blood B cells, but not in high density sIgM+(sIgG-) B cells prepared with Percoll density gradients and magnetic beads separation techniques; the latter cells were therefore used throughout this study . Induction of germ-line transcripts (germ-line C gamma 1 transcript) was noted in stimulated B cells (SAC plus IL-2) at 6 h; thus, the appearance of germ-line transcripts could not be attributed to preferential growth of B cells expressing germ-line transcripts . In subsequent studies we found that induction of germ-line transcripts for the various IgG subclasses fell into two patterns . Induction of C gamma 3 and C gamma 1 germ-line transcripts, transcripts of genes in the first human duplication unit, generally required a proliferative stimulus (Staphylococcus aureus Cowan I, SAC) and was brought about by SAC plus IL-4 (C gamma 3 germ-line transcripts) and SAC alone or SAC plus IL-2 (C gamma 1 germ-line transcripts); in contrast, induction of C gamma 2 and C gamma 4 germ-line transcripts, transcripts of genes in the second duplication unit, was accomplished with cytokines alone: IFN-gamma (C gamma 2 germ-line transcripts) and IL-4 (C gamma 4 germ-line transcripts), and was not augmented by addition of a proliferative stimulus . Finally, we found that IFN-gamma inhibited IL-4-induced C gamma 3 and C gamma 4 germ-line transcripts (with or without SAC) . These findings establish that the various human IgG subclasses manifest distinct requirements for the regulation of early steps in isotype differentiation . In addition, they suggest that human C gamma genes exhibit patterns of regulation related to their respective duplication units. Ann Intern Med, 1993 Oct 1, 119(7 Pt 1), 560 - 7 Prosthetic valve endocarditis resulting from nosocomial bacteremia . A prospective, multicenter study; Fang G et al.; OBJECTIVE: To determine the incidence of endocarditis in bacteremic patients with prosthetic heart valves and the risk factors for and the effect of duration of antibiotic therapy on development of endocarditis in such patients . DESIGN: Multicenter, prospective observational study . SETTING: Six university teaching hospitals with high-volume cardiothoracic surgery . PARTICIPANTS: One hundred seventy-one consecutive patients with prosthetic heart valves who developed bacteremia during hospitalization . MEASUREMENTS AND MAIN RESULTS: Patients were evaluated when they were identified as having bacteremia and 1, 2, 6, and 12 months after its occurrence . Of 171 patients, 74 (43%) developed endocarditis: Fifty-six (33%) had prosthetic valve endocarditis at the time bacteremia was discovered ("endocarditis at outset"), whereas 18 (11%) developed endocarditis a mean of 45 days after bacteremia was discovered ("new endocarditis") . Mitral valve location and staphylococcal bacteremia (Staphylococcus aureus or S . epidermidis) were significantly associated with the development of "new" endocarditis . All 18 cases of new endocarditis were nosocomial, and in 6 of these cases (33%) bacteremia was acquired via intravascular devices . Twenty-one patients without evidence of endocarditis at the time of bacteremia received short-term antibiotic therapy (< 14 days); 1 patient (5%) developed endocarditis . Eleven of 70 patients (16%) who received long-term antibiotic therapy (> 14 days) developed endocarditis (P > 0.2) . CONCLUSIONS: Bacteremic patients with prosthetic heart valves were at notable risk for developing endocarditis, even when they received antibiotic therapy before endocarditis developed and regardless of the duration of such therapy . Intravascular devices were a common portal of entry. Ostomy Wound Manage, 1993 Oct, 39(8), 26 - 7, 30, 32 Wound cultures: what, when and how; Burdette-Taylor S et al.; Collecting wound cultures is a very controversial issue at this time when cost containment is at the forefront of healthcare providers' minds . It is also controversial in this time of Methicillin Resistant Staphylococcus-aureus (MRSA) becoming more common and very difficult to manage in acute and long term care facilities . While the basics of culturing may have not changed, the decision to culture or not, and knowing what, when and how to culture, are of vital importance in the treatment and quality of care for the patient, staff and others . The purpose of this article is to present the implications for culturing a wound and the proper procedure for collecting the specimen and planting the organism . In this discussion of wound culturing, the authors will provide guidelines to accurately interpret the results to accommodate proper treatment in a cost-effective manner. Mech Ageing Dev, 1993 Oct 1, 71(1-2), 31 - 46 Age-related alterations in the activation and expression of phosphotyrosine kinases and protein kinase C (PKC) among human B cells; Whisler RL et al.; Age-related changes in the functional properties of human B cells have been reported by several groups, but little is known about the early biochemical events and signaling pathways that might be altered during aging . The present investigation examined whether differences in the activation of protein tyrosine kinases (PTK) and in the expression of protein kinase C (PKC) enzymatic activity could be identified in B cells from 16 elderly subjects (mean 77 years) compared to B cells from 15 young subjects (mean 33 years) . B cells from young subjects stimulated with the surface immunoglobulin (sIg) crosslinkers anti-IgM or Staphylococcus aureus Cowan I (SAC) demonstrated rapid increases in PTK mediated de novo tyrosine phosphorylation of endogenous proteins . In comparison, stimulated B cells from elderly subjects were reduced 22-46% in tyrosine phosphorylations . Quantitation of the enzymatic levels and activation/translocation of PKC activity among resting and sIg stimulated B cells showed that B cells from approximately 50% of elderly subjects had significant reductions compared to B cells from young subjects . Further analyses of the expression of PTK and PKC enzymatic activity by stimulated B cells from elderly subjects demonstrated that aging was associated with greater heterogeneity in PKC expression and that defects in PKC enzymatic activity could coexist with relatively normal PTK activity . Thus, these data suggest that aging can alter the expression of PTK/PKC enzymatic activity in human B cells and that these age-related alterations might perturb the balance between PKC-dependent and -independent signaling pathways. Eur J Clin Microbiol Infect Dis, 1993 Oct, 12(10), 769 - 72 Non-tropical pyomyositis in adults: report of four cases and literature review; Skoutelis A et al.; Four cases of non-tropical pyomyositis in adults are presented and 72 cases reported in the literature are reviewed . The diagnosis is often overlooked or delayed because most physicians are not familiar with the entity . Local signs of inflammation fever, leukocytosis and an elevated erythrocyte sedimentation rate are common features . Staphylococcus aureus is the most common pathogen and the thigh muscles are the most common site involved . Computerized tomography is used to establish the diagnosis and surgical incision and drainage in combination with antibiotic therapy is successful in the majority of the cases. Clin Investig, 1993 Oct, 71(10), 780 - 6 Iatrogenic epidural spinal abscess; Bollensen E et al.; We present four cases of iatrogenic epidural spinal abscess directly caused by externally introduced catheters or probes . In two patients the infection spread per continuum, in the other two patients due to haematogenous dissemination . Clinical presentation in each case included generalized malaise with fever, signs of meningeal inflammation and focal neurological signs at the spinal level . The diagnosis was made on the basis of inflammatory changes in the cerebral spinal fluid and localization of the abscess by means of computer and magnetic resonance tomography . A broad-spectrum antibiotic regimen included a penicillinase-resistant preparation because of the frequent involvement of Staphylococcus aureus . It is our experience that a good outcome is dependent on early and specific treatment. Comp Immunol Microbiol Infect Dis, 1993 Oct, 16(4), 323 - 30 Antibody dependent haemolysin, complement and opsonin in sera of a major carp, Cirrhina mrigala and catfish, Clarias batrachus and Heteropneustes fossilis; Saha K et al.; The present communication is a continuation of our earlier study on the natural serum haemagglutinin/lectins of Cirrhina mrigala, Clarias batrachus and Heteropneustes fossilis . Sera of Cirrhina mrigala, belonging to the major carp family, could not only agglutinate heterologous rabbit erythrocytes, but also lyse them spontaneously . This lysis of rabbit RBC by Cirrhina mrigala sera was calcium ion dependent and heat sensitive, indicating thereby that the haemolysis was mediated by the fish serum complement system via the classical pathway . Quantification of CH50 and APCH50 levels in the sera of Clarias batrachus and Heteropneustes fossilis as well as in the sera of amphibia, aves and mammals showed that lower vertebrates predominantly possessed an alternative pathway of the complement system, while on the other hand, in the higher vertebrates the major pathway of complement activation was classical . Furthermore sera of Clarias batrachus and Heteropneustes fossilis had opsonins, which could stimulate heterologous rat peritoneal macrophages to engulf Staphylococcus aureus with the production of superoxide anion . From this study we concluded that fishes have been armed with various powerful natural humoral defense systems for their protection against environmental pathogens. Comp Immunol Microbiol Infect Dis, 1993 Oct, 16(4), 317 - 21 Studies on some biochemical characteristics of Staphylococcus aureus of buffalo mammary origin; Varshney JP et al.; A total of 49 isolates of Staphylococcus aureus of buffalo mammary origin were studied for biochemical characteristics . Coagulase production, clumping factor, haemolytic activity, pigment production and fermentation of maltose and mannitol were employed to differentiate S . aureus from S . hyicus and S . intermedius . Out of 49 isolates, 97.95, 93.87, 93.87, 89.79, 95.91, 100.0, 95.91, 59.18, 100.0, 100.0, 100.0, 89.79, 91.83 and 100.0% isolates were positive for coagulase production, protein-A production, haemolysin production, thermostable nuclease production, deoxyribonuclease production, tellurite reduction, nitrate reduction, lipase production, phosphatase production, mannitol fermentation, glucose fermentation, M.R . test, V.P . test and pigment production respectively . The only isolate from which coagulase production could not be detected, however, showed haemolytic activity, protein-A productivity, pigmentation and mannitol fermentation . One of the protein-A negative isolate was coagulase positive and showed mannitol fermentation, pigmentation and haemolytic activity . The study revealed that the biochemical characteristics of S . aureus of buffalo mammary origin did not differ from those of cattle origin . Coagulase, haemolysin, thermostable nuclease, deoxyribonuclease, phosphatase, lipase, tellurite and nitrate reduction closely related with protein-A . The presence of protein-A seems to be as reliable an indicator for S . aureus of buffalo origin as is coagulase production. APMIS, 1993 Oct, 101(10), 753 - 61 Comparative study of the cellular fatty acids of methicillin-resistant and -susceptible Staphylococcus aureus; Asai S et al.; The cellular fatty acid compositions of 26 strains of methicillin-resistant Staphylococcus aureus (MRSA) and 17 strains of methicillin-susceptible S . aureus (MSSA) were analyzed by gas-liquid chromatography . The fatty acid compositions of the two groups were very similar with 16 identified components . The major fatty acids were Ci14 = 0, Ci15 = 0, C18 = 0 and C20 = 0 . Among these fatty acids, the percentage of the Ci15 = 0 fatty acid component of MRSA strains (11.4 +/- 3.9%) was statistically higher than that of MSSA strains (6.2 +/- 2.4%) (p < 0.001) . On the other hand, the percentage of the C20 = 0 fatty acid components of MRSA strains (20.2 +/- 8.8%) was statistically lower than that of MSSA strains (30.7 +/- 10.4%) (p < 0.001) . The production of beta-lactamase and beta-hemolysin in both groups' strain was also unrelated to the relative amounts of the fatty acid components . These results indicated a statistical tendency for the percentage fatty acid compositions of the MRSA strains to be quantitatively different from those of the MSSA for both the Ci15 = 0 and C20 = 0 fatty acid components . Analysis of the fatty acid compositions may have an application in the differentiation of MRSA and MSSA strains. Int J Immunopharmacol, 1993 Oct, 15(7), 829 - 32 IL-4 receptor expression by SAC-activated B-lymphocytes: its role in B-cell proliferation and the effect of cyclosporine (CsA), prednisolone and verapamil; Degiannis D et al.; Expression of the IL-4 receptor was studied in a highly purified population of human B-lymphocytes stimulated by Staphylococcus aureus, cowan I (SAC) . Flow cytometric analysis showed that incubation with SAC in the absence of detectable levels of IL-2, IL-4 and IL-6 resulted in a striking increase in cellular binding of IL-4 . The SAC-stimulated B-cells responded to exogenous IL-4 by DNA synthesis . This response was unaffected by CsA or prednisolone, but was inhibited by the Ca2+ channel blocker verapamil. Arch Dis Child, 1993 Oct, 69(4), 443 - 5 Resolution of hepatic abscess after interferon gamma in chronic granulomatous disease; Hague RA et al.; Recombinant interferon gamma has been used prophylactically in children with chronic granulomatous disease, but its role in the treatment of acute infective episodes has not been defined . A 3 year old boy presented with multiple candidal liver abscesses and was given intravenous antifungal treatment and he showed initial improvement . After six weeks his erythrocyte sedimentation rate and C reactive protein remained raised, and a computed tomogram showed a single abscess in the left lobe of the liver from which pus was drained and Staphylococcus aureus isolated . During the next eight months the abscess persisted despite appropriate intravenous antibiotics and percutaneous drainage . Subphrenic extension precluded definitive surgery . Nine months after initial presentation recombinant interferon gamma 0.05 mg/m2 intravenously was commenced three times a week . Complete resolution occurred within two months . It is concluded that interferon gamma is useful in treating infective episodes, and further study of the use of prophylactic antimicrobial treatment and intermittent interferon gamma during acute episodes is now required. Clin Ter, 1993 Oct, 143(4), 279 - 90 Sepsis-bacteraemia and other infections due to non-opportunistic bacterial pathogens in a consecutive series of 788 patients hospitalized for HIV infection; Manfredi R et al.; A retrospective study was carried out to evaluate the incidence, etiology and role of non-opportunistic bacterial infections in a series of 788 consecutive HIV-infected patients hospitalized during the last 7 years; 71.9% of patients were i.v . drug addicts, 15.6% homo-bisexual men, 7.4% heterosexuals, 2.7% haemophiliacs and 2.4% children with vertically-acquired infection . According to the CDC classification of HIV infection, 71 patients were classified as CDC I-II, 148 as CDC III, and 569 (72.2%) as CDC IV . Diagnosis of bacterial infection was based on signs and symptoms, in association with the isolation of microorganisms consistent with the clinical picture . Two hundred and nineteen patients out of 788 (27.8%) (4 in CDC group I-II, 34 in CDC III and 181 in CDC IV), presented one or more episodes of non-opportunistic bacterial infection . The morbidity of these infections showed a significant correlation with the progression of HIV disease (CDC III vs . CDC I-II {p < 0.003} and CDC IV vs . CDC III {p < 0.05}), but no significant relation was found with age, sex or type of risk for HIV infection . The most frequent clinical picture was sepsis/bacteraemia (33.3%), followed by respiratory tract (23.8%), and genitourinary tract (20.8%) infections . A total of 399 bacterial pathogens were isolated in 303 different episodes of infection: 211 (52.9%) were gram-positive and 188 gram-negative, with Staphylococcus aureus (69 isolations), Staphylococcus epidermidis (50), and Pseudomonas spp . (48) as the major pathogens . Sepsis/bacteraemia was the most frequent clinical picture associated with growth of gram-positive pathogens, while detection of gram-negative bacteria appeared more significantly related with genitourinary or respiratory tract localizations . Bacterial infections in hospitalized HIV-infected patients, even though rarely life-threatening, need particular attention because of the high incidence and recurrence rate, the wide aetiological and clinical spectrum, the frequent microbial associations and the strict relationship with the progression of HIV disease. Rev Clin Esp, 1993 Oct, 193(5), 235 - 8 {Pyogenic sacroiliitis . Presentation of 10 cases}; Gutierrez Macias A et al.; We report the clinical, radiological and microbiological features of ten patients diagnosed of pyogenic sacroiliitis in an Internal Medicine Department during a 13 years period . Clinical signs and radiological or scintigraphical findings were present in every case . Staphylococcus aureus was isolated in blood cultures in eight cases . We remark mean age (26.2 years), male predominance and the high prevalence of intravenous drug abuse as the main epidemiological data in our patients . Onset was acute in 60% of the cases, and subacute in the others . Erythrocyte sedimentation rate and a high white blood cell count were uniformly elevated among laboratory tests . Initial roentgenograms were normal in 60% . Bone scans were the most sensitive diagnostic aid . All the patients were treated with intravenous cloxacilin between two and eight weeks; aminoglycosides were added in four patients . We have observed two cases of psoas abscesses that required surgical drainage. Kansenshogaku Zasshi, 1993 Oct, 67(10), 987 - 91 Nasal carriers of Staphylococcus aureus among healthy individuals; Tanaka Y et al.; Staphylococcus aureus and Staphylococcus spp . were isolated from healthy students and their drug resistance was investigated . S . aureus was isolated from 17 of 70 persons (24.3%) . Fourteen strains of 22 isolates of S . aureus were ampicillin-resistant . Two strains each were cefmetazole-resistant and gentamicin-resistant . None of the strains was found to be methicillin-resistant . Compared with the strains isolated from the hospital ward environment, S . aureus in healthy individuals was relatively sensitive to antibiotics. Kansenshogaku Zasshi, 1993 Oct, 67(10), 1005 - 10 {Methicillin resistance, production of toxic shock syndrome toxin-1 and types of enterotoxin, protease and coagulase at vaginal isolates of Staphylococcus aureus}; Arai K et al.; The vaginal colonization with Staphylococcus aureus, including strains that have methicillin resistance and production of toxic shock syndrome toxin-1 (TSST-1), enterotoxins and protease was studied in 308 women at the time of delivery . Thirty-six of these women were colonized with S . aureus in the vagina . All of the isolates were methicillin sensitive S . aureus (MSSA), except one women who had both methicillin resistant S . aureus (MRSA) and MSSA . The strains, included MRSA, did not elaborate the TSST-1 and produced protease . The distribution of protease pattern (types) observed by MSSA strains on standard methods caseinate agar plates was type A, 1 strain; type B, 15 strains; type C, 6 strains; type D, 14 strains . Sixteen of the 36 strains produced non of the enterotoxins and 20 strains produced enterotoxins and the type distribution was enterotoxin type A, 6 strains; type B, 10 strains; type C, 1 strain; A and B, 1 strain; A, B and C, 2 strains . These results provide the possibility of pathogenic S . aureus colonized in vagina transmit to neonates at the time of delivery. Int J Food Microbiol, 1993 Oct, 20(1), 45 - 8 Growth of Staphylococcus aureus in pasta in relation to its water activity; Valik L et al.; Water activity (aw) of pasta decreased linearly with time during drying . A strain of Staphylococcus aureus isolated from commercial pasta began multiplying immediately after inoculation but decreased in number when aw was reduced to below 0.93 . Another strain of S . aureus appeared to multiply until the aw was below 0.86 . The results are discussed in relation to practices for pasta production. Appl Environ Microbiol, 1993 Oct, 59(10), 3327 - 33 Rapid, automated separation of specific bacteria from lake water and sewage by flow cytometry and cell sorting; Porter J et al.; The use of fluorescence-activated flow cytometric cell sorting to obtain highly enriched populations of viable target bacteria was investigated . Preliminary studies employed mixtures of Staphylococcus aureus and Escherichia coli . Cells of S . aureus, when mixed in different proportions with E . coli, could be selectively recovered at a purity in excess of 90% . This was possible even when S . aureus composed only approximately 0.4% of the total cells . Cell sorting was also tested for the ability to recover E . coli from natural lake water populations and sewage . The environmental samples were challenged with fluorescently labelled antibodies specific for E . coli prior to cell sorting . Final sample purities of greater than 70% were routinely achieved, as determined by CFU . Populations of E . coli released into environmental samples were recovered at greater than 90% purity . The use of flow cytometry and cell sorting to detect and recover viable target bacteria present at levels of less than 1% within an indigenous microflora was also demonstrated. Int J Oral Maxillofac Surg, 1993 Oct, 22(5), 292 - 7 Septic arthritis of the temporomandibular joint: review of the literature and report of two cases in children; Leighty SM et al.; Septic arthritis of the temporomandibular joint (TMJ) has a high morbidity, is infrequently reported, and has been described almost exclusively in adults . We present two cases of septic arthritis of the TMJ that occurred in children after minor blunt trauma . Literature related to septic arthritis of the TMJ was reviewed, and a composite list of cases was constructed . The most common causes were various infections of the head and neck, rheumatic joint disease, and iatrogenesis . Pathogens may gain access to the TMJ by several routes . Patients typically present with an acute, tender, monarticular arthritis with associated swelling and erythema . Malaise, nausea, and vomiting may also be present . Traumatic effusions, fractures, and neoplasms may present in a similar fashion, and mimic TMJ septic arthritis . Staphylococcus aureus is the most commonly reported pathogen and often causes permanent joint damage . Aspiration and analysis of joint fluid, as well as blood chemistry, imaging studies, and clinical impression, may assist in the diagnosis . Timely diagnosis and treatment are essential for a successful outcome; therapy should include antimicrobial agents, adequate drainage, and resting of the joint . Complications include spread of infection, postinfectious bony changes, and fibrous (or bony) ankylosis of the temporomandibular joint. Cytometry, 1993 Oct, 14(7), 811 - 8 Rapid flow cytometric antibiotic susceptibility assay for Staphylococcus aureus; Ordonez JV et al.; Conventional techniques for antibiotic susceptibility testing usually require 24 h or more to produce accurate results . This long waiting period to demonstrate antibiotic action is necessary because such tests rely on growth (or the lack of it), when the microorganisms are incubated in the presence of the antibiotic . In an effort to improve antibiotic susceptibility testing, we developed a flow cytometric technique for Staphylococcus aureus in which antibiotic action is determined by monitoring drug-induced membrane potential changes, using the potential-sensitive dye 3,3'-dipentyloxacarbocyanine iodide . Three ATCC reference strains of S . aureus and 13 unknown strains of the same microorganism were tested for susceptibility to penicillin G and oxacillin . Our results indicate that susceptibility of S . aureus to these antibiotics can be measured reliably at 90 min after addition of the antibiotic, and the results are comparable to those obtained with conventional susceptibility tests. Circ Shock, 1993 Oct, 41(2), 88 - 102 Retrospective study comparing the pathophysiology of antibiotic-treated and untreated Escherichia coli- and Staphylococcus aureus-infused baboons; He S et al.; Escherichia coli and Staphylococcus aureus are the most common pathogens encountered in septic shock . This is a descriptive study in which the pathophysiologic response to infusions of LD100 concentrations of E . coli and S . aureus are staged and compared . Equivalent concentrations of both organisms were infused over a 2 hr period into antibiotic-treated and untreated animals with the following results: 1) The apparent clearance of E . coli was less than that of S . aureus over the 2-hr infusion period, but far greater during the next 8 hr in both antibiotic-treated and untreated animals . Thus the clearance of E . coli fits a one-compartment (intravascular), and that of S . aureus fits a two-compartment (intra- and extravascular) model . 2) The intensity of the cardiovascular, temperature, and metabolic response to E . coli was greater, whereas that of the disseminated intravascular coagulant (DIC) response to S . aureus was greater . We conclude, therefore, that the response to E . coli consists of four stages with no invasion and colonization of tissues, whereas the response to S . aureus consists of two stages with invasion and colonization of tissues. Allergy, 1993 Oct, 48(7), 504 - 10 Effects of interleukin-4 and interferon-gamma on the secretion of IgG4 from human peripheral blood mononuclear cells; Sutherland M et al.; Whereas IgE antibodies are linked with allergy, IgG4 antibodies may reflect the state of immunity and protection against a particular antigen . It has been shown that interleukin (IL)-4 is required for induction of IgE synthesis . In order to elucidate the role of IL-4 in the production of IgG4 and to compare IgG4 and IgE regulatory processes, we quantified these immunoglobulin isotypes after in vitro culture of peripheral blood mononuclear cells (PBMC) in the presence of IL-4 . The production of IgG4 was increased by IL-4 under the same conditions which are optimal for IgE production but not among PBMC from all donors, depending on the magnitude of spontaneous IgG4 secretion: IL-4 was effective only when the spontaneous secretion of IgG4 was < 7% of the total IgG secretion; it had no effect when spontaneous IgG4 production was > 7% of total IgG . The IL-4-induced IgE response was consistently obtained when IgG4 was < 7% of total IgG but was markedly diminished or absent when IgG4 was > 7% of total IgG . If Staphylococcus aureus strain Cowan 1 (SAC) was present during the 48-h preincubation step, spontaneous IgG4 production was increased, but the stimulatory effect of this mitogen on immunoglobulin production, including IgG4, was markedly blocked by the addition of IL-4 . In contrast, IL-4-induced IgE synthesis was strongly blocked by the presence of SAC . Finally, secretion of IgG4 (spontaneous and IL-4-induced) was suppressed among cells from most donors by interferon-gamma (IFN-gamma).(ABSTRACT TRUNCATED AT 250 WORDS) Am J Physiol, 1993 Oct, 265(4 Pt 1), C927 - 33 Metabolism of a long-chain diacylglycerol by permeabilized A10 smooth muscle cells; Chuang M et al.; The regulatory effects of diacylglycerol (DAG) second messengers will be terminated by metabolism . A long-chain DAG, 1-palmitoyl-2-{1-14C}oleoyl-sn-glycerol (2-{14C}POG), was metabolized by cultured A10 smooth muscle cells after permeabilization by preincubation with 340 U/ml alpha-toxin from Staphylococcus aureus . In contrast to results with the cell-permeable DAG analogue, dioctanoyl-glycerol ({3H}diC8), no appreciable 2-{14C}POG degradation could be detected in control A10 cells not treated with alpha-toxin . With permeabilized A10 cells, 2-{14C}POG was mainly converted into lipolytic products of a lipase pathway, monoacylglycerol (MG) and fatty acid (FA); very little radioactivity was incorporated into triacylglycerol (TG) or phospholipid (PL) via reactions catalyzed by either DAG acyltransferase, cholinephosphotransferase, or DAG kinase . Similar results were obtained in experiments with 1-stearoyl-2-{1-14C}arachidonoyl-sn-glycerol . The conversion of 2-{14C}POG into PL and TG was not enhanced by the addition of 1 mM ATP-MgCl2, 1 mM CDP-choline, or 1 mM oleoyl-CoA to the alpha-toxin-treated A10 cells . The formation of FA and MG by permeabilized A10 cells was inhibited by DAG lipase inhibitors, U-57,908 (50 microM) and tetrahydrolipstatin (1-25 nM) . The predominant contribution of the lipase pathway to the metabolism of a long-chain DAG, 2-{14C}POG, by alpha-toxin-treated A10 cells is similar to results for the degradation of {3H}diC8 by intact A10 cells. Mol Gen Genet, 1993 Oct, 241(1-2), 185 - 92 Characterization of the Staphylococcus aureus chromosomal gene pcrA, identified by mutations affecting plasmid pT181 replication; Iordanescu S; The Staphylococcus aureus chromosomal gene pcrA, identified by mutations, such as pcrA3, that affect plasmid pT181 replication, has been cloned and sequenced . The pcrA gene encodes a protein with significant similarity (40% identity) to two Escherichia coli helicases: the helicase II encoded by the uvrD gene and the Rep helicase . The pcrA3 mutation was found to be a C to T transition leading to a threonine to isoleucine substitution at amino acid residue 61 of the protein . The pcrA gene seems to belong to an operon containing at least one other gene, tentatively named pcrB, upstream from pcrA . The PcrA protein was shown to be essential for cell viability and overproduction has deleterious effects on the host and plasmid replication. Kyobu Geka, 1993 Oct, 46(11), 956 - 9 {Successful treatment by using a pedicled omental flap for sternal osteomyelitis and mediastinal infection caused by MRSA}; Morimoto T et al.; A 56-year-old male case, who was complicated with sternal osteomyelitis and mediastinitis after aortic valve replacement, was reported . Its infection was caused by MRSA (methicillin resistant Staphylococcus aureus) . He was treated with sterilization of wound using povidone iodine, Minomycin solution and its injection for 10 days . And thereafter we decided to apply the plastic surgical technique to this case . After debridement of the infected granulation and sternum thoroughly, a pedicled omental flap was transposed successfully. Infect Control Hosp Epidemiol, 1993 Oct, 14(10), 576 - 8 Infection with methicillin-resistant Staphylococcus aureus among hospital employees; Muder RR et al.; OBJECTIVE: To describe the spectrum of clinical infection caused by methicillin-resistant Staphylococcus aureus (MRSA) in healthcare workers . DESIGN: Case series . SETTING: Two Veterans Affairs hospitals in which methicillin-resistant S aureus (MRSA) is endemic . PATIENTS: Five employees presenting to employee health or infectious disease clinic . RESULTS: All employees had had direct exposure to patients colonized with MRSA . Employee infections included cellulitis, impetigo, folliculitis, paronychia, and conjunctivitis . MRSA was isolated from all clinically infected sites and from the anterior nares of two employees . Three employees received a variety of ineffective oral antimicrobials before MRSA was recognized as the causative agent . All infections responded to appropriate therapy . CONCLUSIONS: Employees of hospitals with endemic MRSA may acquire MRSA infection . Presentation in our employees was that of relatively uncomplicated soft tissue infection, but several employees received inappropriate therapy before bacteriologic diagnosis . We recommend that culture and susceptibility testing be obtained prior to institution of therapy when hospital employees present with soft tissue infection. J Dairy Sci, 1993 Oct, 76(10), 2913 - 24 Histologic response of the heifer mammary gland to intramammary infusion of interleukin-2 or interferon-gamma; Quiroga GH et al.; The histologic and cytologic responses of heifer mammary glands infected with Staphylococcus aureus were studied after infusion of interleukin-2 or interferon-gamma . Two groups of 4 heifers each, which were infected experimentally with S . aureus in all 4 mammary quarters, were infused in diagonal quarters with 7.5 x 10(5) units of interleukin-2 or 10(5) units of interferon-gamma; remaining quarters received PBS . Heifers in both trials were slaughtered 14 d after cytokine infusion, and mammary tissues were collected for histological examination . Uninfected quarters from 2 additional heifers were left untreated to compare infected with uninfected tissues for both trials . Morphologic evaluation and leukocyte infiltration scores were performed on tissue sections stained with hematoxylin and eosin, and plasma cells were quantified on sections stained with immunoperoxidase . Infected quarters had lower percentages of alveolar epithelial and luminal areas and higher percentages of stromal area than did uninfected quarters in the interleukin-2 trial, but no differences were observed between infected quarters that had been treated with PBS or interleukin-2 . Likewise, interferon-gamma treatment had no effect on mammary parenchymal components in the infected quarters . Interleukin-2 treatment significantly elevated leukocytosis into the mammary gland parenchyma compared with infected quarters treated with PBS and uninfected quarters . Among the leukocyte types evaluated, eosinophilic infiltration was elevated in interleukin-2 quarters over that of PBS controls . In both trials, concentrations of plasma cells bearing Ig were elevated in infected versus uninfected quarters . Plasma cell concentrations also were higher in cytokine than PBS controls, especially in interleukin-2 quarters . Results suggested that neither cytokine influenced the histology of infected mammary tissues, but both interleukin-2 and interferon-gamma increased, although insignificantly, the prevalence of all isotypes of plasma cells bearing Ig, suggesting enhancement of the local immune response to IMI. J Clin Periodontol, 1993 Oct, 20(9), 662 - 7 Late onset Papillon-Lefèvre syndrome? A chromosomic, neutrophil function and microbiological study; Bullon P et al.; We had the opportunity to study a family with one of the most destructive forms of periodontal disease known, the Papillon-Lefevre syndrome . The parents had no consanguinity and were not affected, and were therefore to be considered carriers of the disease . 2 sisters, the eldest and youngest, showed periodontal breakdown and hyperkeratotic skin lesions, but their deciduous dentition was not affected . 2 brothers had skin lesions only and another brother and sister were healthy . Furthermore, 2 babies died at birth one after a 9-month pregnancy and the other after a 6-month pregnancy, and the mother also suffered 3 miscarriages . For 4 years, we studied the family: in the case of both sisters, mechanical periodontal treatment and antibiotics were unable to control the disease . In the chromosomic study of the 2 sisters affected, the GTG banding technique found no trace of anomalies in the cells analyzed, whose chromosomic formation was 46,XX . Before treatment, the chemotaxis of the PMN, the phagocytosis of opsonized Staphylococcus aureus, and production of superoxide radicals by PMN was significantly impaired in both sisters . Despite scaling and root planing, the periodontal lesions still progressed, but the PMN functions evaluated were now normal in both sisters . An orally asymptomatic but dermatologically affected brother showed no significant defect in the phagocytic activity and the production of superoxide radicals. Int J Biochem, 1993 Oct, 25(10), 1399 - 406 Collapse of K+ and ionic balance during photodynamic inactivation of leukemic cells, erythrocytes and Staphylococcus aureus; Malik Z et al.; 1 . The immediate and fast ionic fluxes in Friend erythroleukemic cells (FELC), erythrocytes and Staphylococcus aureus during short intervals of porphyrin mediated photosensitization were determined uniquely by X-ray microanalysis (XRMA) combined with electron microscopy . 2 . Photodynamic inactivation of FELC was mediated by either endogenous protoporphyrin induced by 5-amino levulinic acid (5-ALA), or Photofrin-II . We describe the predominant phenomena of > 85% K-loss within 2-10 min of photoactivation . However the accompanied Na inflow and the collapse of the cellular balance of elemental-composition were inconsistent and acted as a function of cell damage . 3 . Erythrocytes treated with hematoporphyrin (HP) lost most of their intracellular K yet instantly gained Na . Nevertheless the K/Na molar ratio of the control erythrocytes was nearly 12/1 while after photosensitization and K loss it changed to 1/1 . 4 . The S . aureus bacteria photosensitized with HP showed entire K-loss as well as marked Na efflux which increased with irradiation time; this was accompanied by the decline of other cell elements . 5 . The prevailing K loss in FELC, erythrocytes and bacteria during the first minutes of photosensitization is deduced to be an immediate primary consequence of the photodynamic effect, while other ionic changes are joined in order with the development of cellular damage. Anesth Analg, 1993 Oct, 77(4), 766 - 8 Growth of Staphylococcus aureus in four intravenous anesthetics; Sosis MB et al.; Patient infections related to the use of propofol have been reported . To investigate the growth of Staphylococcus aureus in propofol, thiopental, methohexital, etomidate, and 0.9% saline containing no bacteriostatic drug, these preparations were inoculated and samples were plated onto blood agar at 0, 3, 6, 21, 24, and 27 h . The number of colony-forming units (CFU) on the plates was then determined after 24 h of incubation . Samples from the inoculated etomidate solution showed zero CFU at 3 h and thereafter, whereas 21 h were required by the methohexital and thiopental solutions to reduce the number of CFU to zero . For normal saline, no significant change in CFU was seen before the first 6 h, then the number of CFU gradually declined, although some S . aureus CFU were still present at 27 h . Inoculation of the propofol emulsion resulted in a substantial growth of S . aureus between 6 and 21 h after inoculation . We conclude that, of the preparations tested, only propofol was an excellent medium for the rapid growth of S . aureus . Meticulous sterile technique, therefore, is advised when handling it. South Med J, 1993 Oct, 86(10), 1175 - 8 Staphylococcus aureus meningitis associated with pyogenic infection of the sacroiliac joint; Cieslak TJ et al.; Although we have reported the first case of Staphylococcus aureus meningitis due to pyogenic arthritis of the sacroiliac joint, this finding is actually not surprising, given the strong association between this form of meningitis and underlying bone, joint, and soft tissue infections . The physician faced with a case of meningitis due to S aureus in a patient without a history of trauma or neurosurgical manipulation must do a prompt and thorough search for underlying infectious conditions . In the case of sacroiliitis in particular, a high index of suspicion needs to be maintained, given the difficulties and delays in diagnosis associated with this infection . Radionuclide scanning with 99mTc or 67Ga is usually helpful in the early confirmation of this condition, the presence of which may be suspected on the basis of thorough physical examination. Singapore Med J, 1993 Oct, 34(5), 435 - 7 Nasal septal abscess--retrospective analysis of 14 cases from University Hospital, Kuala Lumpur; Jalaludin MA; Fourteen patients who presented to the University Hospital of Kuala Lumpur between June 1981 and June 1991 were reviewed retrospectively . Nasal septal abscesses are uncommon and therefore there are limited reports in the medical literature . Early diagnosis and immediate therapy is mandatory to avoid cosmetic nasal deformity or intracranial infection . Two out of the fourteen patients developed saddle nose deformity and septal perforation because of delay in treatment, the cases were misdiagnosed by non-otolaryngologist as turbinates swelling . The leading cause of nasal septal abscess was non-surgical trauma which accounted for about 85.7% . The commonest pathogenic organism isolated from the pus of nasal septal abscess was Staphylococcus aureus. Zentralbl Veterinarmed B, 1993 Oct, 40(8), 559 - 68 Effect of partially purified Staphylococcus aureus beta-haemolysin on the mammary gland of the mouse; Calvinho LF et al.; Partially purified Staphylococcus aureus Beta-haemolysin and a S . aureus strain (FGH28), isolated from a clinical case of bovine mastitis, known to produce in vitro Beta-, but neither Alpha- nor Delta-haemolysins were each injected separately into the mammary gland of mice . Relative purity and in vitro biological activities of this preparation were assessed . The effects of both the partially purified preparation and the S . aureus strain injected were assessed in terms of mortality, clinical appearance, macroscopic and microscopic changes, recovery of organisms from the inoculated glands and biological activity in supernatants from ground mammary glands . When injected alone, partially purified Beta-Haemolysin was not found to be an important virulence factor, being a diffuse infiltration of neutrophils into the alveoli the pathological effect observed following its inoculation . Vascular lesions were observed, in the most severe cases, following injection of S . aureus FGH28. J Chemother, 1993 Oct, 5(5), 289 - 92 Macrolides induced suppression of virulence factors produced by Staphylococcus aureus; Moneib NA et al.; The effect of sub-MICs of azithromycin, clarithromycin and roxithromycin, as compared to erythromycin, on the production of coagulase, beta-hemolysin, lecithinase and deoxyribonuclease by Staphylococcus aureus was studied . All new macrolides completely inhibited coagulase and beta-hemolysin production and partially inhibited lecithinase and deoxyribonuclease . Such inhibition is not related either to growth inhibition or to inhibition of enzyme activity . Erythromycin, on the other hand, had no effect on coagulase or beta-hemolysin production but slightly suppressed the production of lecithinase and deoxyribonuclease . This inhibitory effect might have clinical significance if it was found to occur in vivo. Mol Microbiol, 1993 Oct, 10(1), 7 - 12 Human Menkes X-chromosome disease and the staphylococcal cadmium-resistance ATPase: a remarkable similarity in protein sequences; Silver S et al.; A search with the proposed amino acid translation product from the new 'candidate gene' for human Menkes disease against protein sequence libraries showed a remarkable similarity to that for the cadmium efflux ATPase from Staphylococcus aureus resistance plasmids . The Menkes sequence appears closer to the CadA Cd2+ sequence than to P-type ATPases from animal sources . Menkes syndrome is an X-chromosome invariably fatal disease that results from aberrant copper metabolism . The gene that is defective in Menkes patients, i.e . the Menkes candidate gene, encodes a P-type ATPase, whose properties satisfactorily explain the phenotype of the disease . P-type ATPases are all cation pumps, either for uptake (e.g . the bacterial Kdp K+ ATPase), for efflux (e.g . the muscle sarcoplasmic reticulum Ca2+ ATPase), or for cation exchange (e.g . the animal cell Na+/K+ ATPase) . These enzymes have a conserved aspartate residue that is transiently phosphorylated from ATP during the transport cycle, hence the name 'P-type' ATPase . The Menkes sequence shares with the staphylococcal CadA ATPase those regions common to all P-type ATPases and also an N-terminal dithiol region that was proposed to be a 'metal-binding motif' . There are one or two copies of this motif in the available CadA sequences and six copies in the Menkes sequence. J Hosp Infect, 1993 Oct, 25(2), 97 - 108 Genetic analysis of community isolates of methicillin-resistant Staphylococcus aureus in Western Australia; Udo EE et al.; Methicillin-resistant Staphylococcus aureus (MRSA) obtained from patients who had not been hospitalized outside Western Australia (WA) were studied for antimicrobial resistance and plasmid content and by pulsed-field gel electrophoresis . They were found to be of several types, none of which appeared to be related to MRSA which have been previously studied . It appears that new MRSA strains have emerged in communities in the far north of WA and are being isolated at an increasing frequency in Perth hospitals 2000 km south. J Hosp Infect, 1993 Oct, 25(2), 91 - 6 Nosocomial spread of Staphylococcus aureus showing intermediate resistance to methicillin; Richards J et al.; A nosocomial outbreak of infection and colonization involving six patients and caused by a strain of Staphylococcus aureus showing intermediate resistance to methicillin (MIC = 4-8 mg l-1) is described . The outbreak was associated with skin-carriage of the epidemic strain by a nurse suffering from severe eczema . The reduced susceptibility of the outbreak strain to methicillin was associated with beta-lactamase production . Elimination or inhibition of beta-lactamase activity produced a two-fold decrease in methicillin MIC . There was no evidence for the presence of either penicillin-binding protein 2a or the corresponding mec gene, which mediate resistance in fully methicillin-resistant strains. Transplantation, 1993 Oct, 56(4), 985 - 90 Responsiveness of preactivated B cells to IL-2 and IL-6 . Effect of cyclosporine and rapamycin; Hornung N et al.; In order to determine which drug may be more effective in clinical abnormalities associated with polyclonal B lymphocyte activation, we compared the in vitro effects of CsA and rapamycin on proliferation or differentiation of preactivated B cells . For that purpose, highly purified B lymphocytes were preactivated in the presence of formalinized Staphylococcus aureus bacteria and then recultured in the presence or in the absence of either rIL-2, rIL-6, or combination or rIL-2 and rIL-6 . After 48 hr in culture, S . aureus bacteria upregulated significantly the binding of phycoerythrin-conjugated IL-2 and IL-6, respectively, by purified B lymphocytes, indicating generation and/or upregulation of receptors for these cytokines . Such preactivated B lymphocytes proliferated in response to optimal concentrations of rIL-2, whereas the addition of rIL-6 to preactivated cells was always accompanied by a decrease of the proliferation rate . CsA upregulated cell proliferation when it was added in the second culture period in the presence or in the absence of rIL-6, whereas rapamycin had no effect in these cases . A combination of rIL-2 plus rIL-6 upregulated significantly the proliferative responses of preactivated B cells . In such cultures both CsA and rapamycin had an inhibitory effect on the proliferative responses . IgM production was unaffected by the addition of rIL-6 to cultures of preactivated B cells, whereas addition of rIL-2 and of the IL-2/IL-6 combination enhanced considerably IgM production . Irrespective of cytokines added, CsA upregulated the production of IgM . In contrast, rapamycin inhibited IgM production in all cases . Our results indicate that, in this experimental system, rapamycin is an effective immunosuppressive agent and its use, at least in vitro, is not accompanied by an upregulation of either the proliferation or differentiation of B lymphocytes. J Clin Invest, 1993 Oct, 92(4), 2053 - 9 Lipopolysaccharide (LPS) recognition in macrophages . Participation of LPS-binding protein and CD14 in LPS-induced adaptation in rabbit peritoneal exudate macrophages; Mathison J et al.; Exposure of rabbit peritoneal exudate macrophages (PEM) or whole blood to picomolar concentrations of LPS induces adaptation or hyporesponsiveness to LPS . Because of the importance of plasma LPS-binding protein (LBP) and the macrophage cell membrane protein CD14 in recognition of LPS, we examined the effect of LBP on LPS-induced adaptation in PEM . PEM exposed to LPS in the presence of LBP for 8 h were markedly less responsive to subsequent stimulation by LPS than monocytes/macrophages (M phi) adapted in the absence of LBP . LPS-induced expression of TNF was sharply reduced in LBP-LPS-adapted PEM, but in contrast these cells remained fully responsive to Staphylococcus aureus peptidoglycan . We considered that specific hyporesponsiveness in LPS-adapted M phi or in blood monocytes could be due to decreased expression of CD14 or diminished binding of LBP-LPS complexes to CD14 . However, flow cytometry analysis revealed only minimal reduction of CD14 expression or CD14-dependent binding of a fluorescent LPS derivative when normo- and hyporesponsive cells were compared . These results show that complexes of LPS and LBP are more effective than LPS alone in inducing adaptation to LPS, and LPS-induced hyporesponsiveness probably results from changes in cellular elements distinct from CD14 that are involved in either LPS recognition or LPS-specific signal transduction. EMBO J, 1993 Oct, 12(10), 3967 - 75 Synthesis of staphylococcal virulence factors is controlled by a regulatory RNA molecule; Novick RP et al.; The production of most toxins and other exoproteins in Staphylococcus aureus is controlled globally by a complex polycistronic regulatory locus, agr . Secretory proteins are up-regulated by agr whereas surface proteins are down-regulated . agr contains two divergent promoters, one of which directs the synthesis of a 514 nucleotide (nt) transcript, RNAIII . In this report, we show that the cloned RNAIII determinant restores both positive and negative regulatory functions of agr to an agr-null strain and that the RNA itself, rather than any protein, is the effector molecule . RNAIII acts primarily on the initiation of transcription and, secondarily in some cases, at the level of translation . In these cases, translation and transcription are regulated independently . RNAIII probably regulates translation directly by interacting with target gene transcripts and transcription indirectly by means of intermediary protein factors. Biochemistry, 1993 Sep 28, 32(38), 9936 - 43 Resolving the two monolayers of a lipid bilayer in giant unilamellar vesicles using deuterium nuclear magnetic resonance; Marassi FM et al.; Giant unilamellar vesicles (GUVs) composed of mixtures of POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) plus DMPG (1,2-dimyristoyl-sn-glycero-3-phosphoglycerol) and/or CHOL (cholesterol) were prepared using detergent dialysis . Vesicles containing at least 30 mol % CHOL had diameters exceeding 450 nm . POPC in such GUVs, deuterium-labeled at either the choline alpha or beta segments, yielded deuterium (2H) and phosphorus (31P) nuclear magnetic resonance (NMR) Pake pattern line shapes, quadrupole splittings and chemical shift anisotropies identical to those obtained with multilamellar vesicles (MLVs) of identical composition . Exposing exclusively the vesicle exterior to either calcium or perchlorate ions, both of which are known to influence lipid head-group conformation through surface charge effects, caused the appearance of two overlapping 2H Pake patterns of equal intensity . The quadrupole splitting of one component remained unchanged while that of the second component was altered in the manner expected for choline alpha or beta deuterons in the presence of a cationic (calcium) or anionic (perchlorate) surface charge . Freeze-thawing the GUVs to equilibrate the exterior and interior vesicular contents eliminated the initially unchanged spectral component . It was likewise possible to resolve two quadrupole splittings when Staphylococcus aureus delta-toxin, a surface-active peptide known to influence lipid head-group orientational ordering, was added to the exterior vesicular solution only . This indicates that delta-toxin upon binding remains confined to one monolayer of the lipid bilayer and does not traverse the membrane.(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1993 Sep 25, 268(27), 20668 - 75 Development of an artificial antibody system with multiple valency using an Fv fragment fused to a fragment of protein A; Ito W et al.; In antigen-antibody interactions, the high avidity of antibodies for their specific antigens can be achieved both by exploiting the high affinity of each binding site and by multivalence of antibodies . In this study, we developed artificial antibodies with multiple valency . The Fv fragments of an antibody fused with one and with two domains of the Fc-binding protein A from Staphylococcus aureus (designated Fv-P and Fv-PP, respectively) were expressed in secreted forms in Escherichia coli . Their physical characteristics were examined . The Fv portions of Fv-P and Fv-PP had virtually the same affinity for the antigen as that of the original Fv molecules, and each protein A-derived domain could be bound to IgG . When Fv-P was mixed with IgG, the complex formed was composed of two Fv-P molecules and one IgG molecule . In the case of Fv-PP, a complex composed of three Fv-PP molecules and two IgG molecules was preferentially formed . Analysis of surface plasmon resonance in a BIAcore system indicated multivalency of these complexes for antigens . There was 3.5-fold decrease in the dissociation constant of the complex between Fv-PP and the antigen upon the addition of IgG . The use of these complexes as reagents in enzyme-linked immunosorbent assay and Western blotting gave much stronger signals than Fv, Fv-P, and Fv-PP alone. J Immunol, 1993 Sep 15, 151(6), 3180 - 6 In vivo tumor immunotherapy by a bacterial superantigen; Ochi A et al.; We have investigated the in vivo efficacy of Staphylococcus aureus enterotoxin B (SEB) coupled to tumor-specific anti-idiotypic antibody in redirecting T cell effector activity to the growth inhibition of B lymphoma 38C13 . Incubation of 38C13 lymphoma cells with syngeneic C3H/He splenic cells and SEB-anti-Id conjugate was associated with between 80 and 100% growth inhibition of the tumor cells . V beta 8+ T cells were integral for the SEB-anti-Id-induced tumor cell growth inhibition . Administration of SEB-anti-Id i.v . to mice previously inoculated with 38C13 lymphoma cells led to greater than 40% survival at 100 days compared to a mean survival of 21 days in control animals . When we compared this reagent with other targeting constructs--the anti-CD3-anti-Id and anti-TCR V beta 8-anti-Id--these more or less effectively prevented tumor growth . However, anti-CD3-anti-Id impaired almost the entire T cell response, whereas the effects of SEB-anti-Id or anti-V beta 8-anti-Id had effects limited to V beta 8+ T cells . Previous studies showed that in vivo administration of SEB caused a small change in V beta 8+ T cell numbers in contrast to anti-V beta 8 antibody, which depleted the entire population . These results together suggest that SEB-anti-tumor antibody conjugates represent a potentially powerful approach for better tumor immunotherapy. JAMA, 1993 Sep 15, 270(11), 1329 - 33 Recurrent epidemics caused by a single strain of erythromycin-resistant Staphylococcus aureus . The importance of molecular epidemiology; Back NA et al.; OBJECTIVE--To compare the accuracy of clinical epidemiologic findings with results of molecular epidemiologic analysis in identifying the source of recurrent epidemics of erythromycin-resistant Staphylococcus aureus (ERSA) infections in a well-baby nursery . DESIGN--Epidemic investigations with retrospective and prospective microbiological surveillance . Staphylococcus aureus isolates were evaluated by plasmid analysis and genotyping . SETTING--A well-baby nursery in a 700-bed university teaching hospital with approximately 250 deliveries per month . PATIENTS--Newborn infants who developed ERSA infections during 1990 and 1991 . INTERVENTION--Traditional infection control measures, including cohorting of infected infants and hand washing, were implemented . Personnel were cultured for nasal carriage of S aureus during both epidemics . Employees carrying ERSA were removed from the nursery and treated with mupirocin . RESULTS--In the first epidemic, 15 infants were infected with ERSA . A nursing assistant who cared for most of the infants was found to be a carrier of ERSA . She was removed from the nursery and the epidemic resolved . Fifteen months after the first epidemic ended, an epidemic that involved 11 infants began . The attending physician had a facial furuncle and was found to be a carrier of ERSA . The physician was treated and the epidemic resolved . Plasmid and genotyping showed the ERSA organisms from both epidemics were the same . The employee implicated in the first epidemic did not have the epidemic strain, but the physician who attended during both epidemics did . CONCLUSIONS--Traditional epidemic investigations may lead to false conclusions that can only be recognized with molecular epidemiologic techniques . For these techniques to be useful in the control of outbreaks, plasmid analysis and/or genotyping must be readily available. JAMA, 1993 Sep 15, 270(11), 1323 - 8 Value of molecular epidemiologic analysis in a nosocomial methicillin-resistant Staphylococcus aureus outbreak; Fang FC et al.; OBJECTIVE--To evaluate two molecular epidemiologic methods used in the analysis of a nosocomial methicillin-resistant Staphylococcus aureus (MRSA) outbreak . DESIGN--Restriction endonuclease analysis of plasmid DNA (REAP) was used in the analysis of 45 MRSA isolates . After termination of the outbreak, isolates were retrospectively analyzed in a blind fashion using the newly described technique of arbitrarily primed polymerase chain reaction (AP-PCR) . Molecular analyses were compared with epidemiologic and antimicrobial susceptibility data . SETTING--Tertiary care university hospital . SUBJECTS--Twenty-eight patients and 12 employees infected or colonized with MRSA during a 6-week period . RESULTS--A clonal relationship demonstrated among isolates from burn unit patients and staff was clearly distinguishable from MRSA isolates arising from other hospital wards . The combination of REAP and AP-PCR provided complementary information in several instances . Aggressive measures to isolate infected patients and eradicate colonization from patients and staff terminated the outbreak . CONCLUSIONS--Although traditional epidemiologic methods retain their central role in modern hospital infection control, molecular epidemiologic analysis can significantly enhance the ability of infection control officers to analyze and terminate hospital epidemics . The combination of AP-PCR and REAP may prove to be a particularly informative means of tracking the nosocomial spread of microbial strains and their mobile genetic elements. J Chromatogr, 1993 Sep 10, 647(1), 3 - 12 Peptide mapping of recombinant human interferon-gamma by reversed-phase liquid chromatography with on-line identification by thermospray mass spectrometry and UV absorption spectrometry; Legrand R et al.; The detection and identification of minor peaks in a complex peptide map of recombinant human interferon-gamma was realized by on-line analysis of the eluted peptides using thermospray mass spectrometry and UV absorbance spectrometry . By this procedure the time-consuming process of collection, purification and chemical sequence analysis is avoided . Owing to the formation of multiple charged ions, the domain of the covered masses is extended . Fragmentation of the peptides in the thermospray source was observed resulting from, amongst others, cleavage by acid hydrolysis of peptide bonds involving an aspartic acid . This was of great use for the identification of peptides in a digest of recombinant human interferon-gamma by Staphylococcus aureus strain V8 endoprotease. J Mol Biol, 1993 Sep 5, 233(1), 170 - 2 Crystallization and preliminary X-ray analysis of a microbial superantigen staphylococcal enterotoxin C2; Passalacqua EF et al.; High yields of staphylococcal enterotoxin C2, from Staphylococcus aureus, have been purified using dye ligand chromatography . Crystals suitable for X-ray diffraction work were obtained by vapour diffusion using ammonium sulphate and polyethylene glycol as precipitants . They belong to the tetragonal space group P4(1)22 with unit cell dimensions a = b = 43.2 A and c = 290.9 A with one molecule per asymmetric unit . The crystals diffract to at least 2.8 A resolution and are suitable for three-dimensional X-ray structural analysis. Pediatr Infect Dis J, 1993 Sep, 12(9), 743 - 7 Role of anaerobic bacteria in liver abscesses in children; Brook I et al.; Aspirates from pyogenic liver abscesses obtained from 14 children were cultured for aerobic and anaerobic bacteria . Of 29 organisms that were recovered, 17 were anaerobic and 12 were aerobic or facultative . The predominant organisms were Peptostreptococcus spp . (5 isolates) . Bacteroides fragilis group (4), Fusobacterium spp . (3) and Staphylococcus aureus (4) . Aerobic or facultative bacteria only were recovered in five cases, anaerobic bacteria only in four and mixed aerobic and anaerobic bacteria in five . Anaerobic bacteria were recovered in liver abscesses that were associated with other infection in which these organisms were predominant (i.e . abdominal infection, abscesses) . This study highlights the potential importance of anaerobic bacteria in pyogenic liver abscess. J Am Dent Assoc, 1993 Sep, 124(9), 76 - 84 Enhancing infection control for elderly and medically compromised patients; Shuman SK et al.; Some patients may be at risk for complications from relatively common infectious diseases . Influenza, tuberculosis and methicillin-resistant Staphylococcus aureus infection can lead to illness and even death in elderly, medically compromised and institutionalized individuals . Dental personnel caring for these individuals should adopt preventive strategies that are simple and inexpensive in addition to standard infection control guidelines. J Clin Microbiol, 1993 Sep, 31(9), 2281 - 5 Phenotypical and genotypical characterization of epidemic clumping factor-negative, oxacillin-resistant Staphylococcus aureus; Schwarzkopf A et al.; A total of 50 oxacillin-resistant Staphylococcus aureus (ORSA) strains that were clumping factor negative (CFN) and protein A negative by latex agglutination were collected from patients in six different hospitals at different locations in Germany during 1991 and 1992 . Antibiograms, bacteriophage typing, and plasmid analysis were performed . The antibiograms showed that, besides oxacillin, all CFN ORSA strains were resistant to gentamicin, clindamycin, erythromycin, ciprofloxacin, and fosfomycin . All these isolates were nontypeable with an international set of phages, and an additional experimental phage set indicated that the strains were phage type 16, 192 . Moreover, all isolates possessed a single plasmid of 30 kb, and restriction analysis of those plasmids revealed identical patterns . For genotyping, these 50 isolates were also analyzed by pulsed-field gel electrophoresis (PFGE) and polymerase chain reaction (PCR) of the coagulase and protein A genes and then by restriction enzyme digestion and analysis of restriction fragment length polymorphisms (RFLPs) . With 49 strains, electrophoresis of SmaI-digested chromosomal DNA revealed identical PFGE patterns regarding the number and size of the DNA fragments, which could be differentiated from those of clumping factor-positive ORSA strains . Typing for the coagulase gene by PCR revealed PCR products of identical sizes . The AluI restriction digestion patterns of the PCR products were identical . PCR with primers derived from the region of that part of the protein A gene that encodes the immunoglobulin G-binding domains showed a PCR product that was about 170 bp smaller than that of the protein A gene from strains that were positive in the protein A latex agglutination test . Since it is precisely this size that is required in order to encode one immunoglobulin G-binding region, we assume that this is not present in the CFN ORSA strains . The phenotypical and genotypical features identify these very unusual CFN ORSA stains as being of clonal origin. FEMS Microbiol Lett, 1993 Sep 1, 112(2), 173 - 7 The antibacterial activity of haemin compared with cobalt, zinc and magnesium protoporphyrin and its effect on potassium loss and ultrastructure of Staphylococcus aureus; Ladan H et al.; The unique antibacterial properties of Fe-protoporphyrin (haemin) on Staphylococcus aureus, compared to Co-protoporphyrin (Co-PP), Mg-protoporphyrin (Mg-PP) and Zn-protoporphyrin (Zn-PP) are described . Only haemin (20 microM) exhibits a strong light-independent antibacterial effect on S . aureus; the other metalloporphyrins, Co-PP, Mg-PP or Zn-PP, have no antibacterial effect in the dark . Only light photosensitization of Mg-PP-treated cells resulted in the inhibition of the bacterial growth, while Co-PP or Zn-PP were photodynamically inactive . A notable effect of haemin on inactivation of S . aureus was the induction of immediate ion fluxes as determined by X-ray microanalysis (XRMA) of fast-frozen cells . A marked efflux of K (96%) and Cl (94%) was expressed immediately as determined by X-ray microanalysis of S . aureus cells treated with haemin for 5 min . Only 48% loss of Na was detected in the cells under these treatment conditions, while P content was increased by 150% . Electron microscopy analysis revealed the appearance of a mesosome-like structure connected to the new septa, filamentous chromosome and arrays of aggregated ribosomes in the cytoplasm . We propose that haemin has multiple cellular targets for its oxidative effect in S . aureus. No Shinkei Geka, 1993 Sep, 21(9), 805 - 8 {Stellate ganglion blocks as the suspected route of infection in a case of cervical epidural abscess}; Makiuchi T et al.; Spinal epidural abscess is a comparatively rare disease . Its prognosis reportedly depends on degree and duration of the neurological symptoms before the treatment . Thus, the importance of early diagnosis and prompt surgical treatment has been emphasized repeatedly . In the case reported here stellate ganglion blocks were considered to be involved in the etiology of a cervical epidural abscess . <Case> The 47-year old woman complained of tinnitus and vertigo and repeatedly underwent stellate ganglion blocks over a period of 10 months . In August, 1991, the patient complained of back pain and developed fever . A few days later she noticed motor weakness and sensory disturbances in the legs . Ten days after the onset of these neurological symptoms she complained of rapidly progressive tetraplegia and was referred to this hospital for admission . On admission, she was fully conscious but febrile . Neurologically, she presented tetraplegia, hypesthesia below level of C7 and slight cervical rigidity . Bladder and bowel dysfunction were also observed . MRI examination showed an epidural mass behind vertebral bodies C6-7 compressing the spinal cord . Antibiotic therapy was initiated immediately and emergency surgical decompression was performed through an anterior approach . Intraoperative findings showed a discitis and yellowish liquid pus in the epidural space . Culture of the pus revealed staphylococcus aureus . In this case repeated stellate ganglion blocks before onset of the symptoms were the suspected route of infection . Postoperative MR images confirmed satisfactory decompression of the spinal cord and motor power was gradually recovered after surgery . Approximately 4 months after surgery she could walk independently . Cervical epidural abscess has been rarely reported as a complication of stellate ganglion block.(ABSTRACT TRUNCATED AT 250 WORDS) J Vasc Surg, 1993 Sep, 18(3), 477 - 84; discussion 485 Direct replacement of mycotic thoracoabdominal aneurysms; Hollier LH et al.; Direct graft replacement with local debridement and prolonged administration of antibiotics was used in the treatment of six patients with mycotic thoracoabdominal aneurysms . The only early death occurred in a patient with systemic sepsis related to Staphylococcus aureus mycotic suprarenal aneurysm . Long-term survival of the remaining patients has been excellent: two patients died of unrelated causes at 5 and 6 years, respectively; one patient remains alive with known persistent infection at 5 years; and the remaining patients are alive with no evidence of infection at 1 1/2 and 10 years, respectively . Percutaneous aspiration of infected perigraft fluid with local instillation of antibiotics along with administration of intravenous antibiotics may provide palliation in selected patients with recurrent infections . In view of the magnitude of the problems associated with recurrent infection, life-time administration of antibiotics is recommended after in situ graft replacement of mycotic thoracoabdominal aneurysms. Eur J Immunol, 1993 Sep, 23(9), 2372 - 4 Linomide, a novel immunomodulator that prevents death in four models of septic shock; Gonzalo JA et al.; Intravenous injections of 50 micrograms Staphylococcus aureus enterotoxin B (SEB) or bacterial lipopolysaccharide (LPS) are lethal, provided that mice are simultaneously sensitized with either N-galactosamine (GalN) or the anti-glucocorticoid RU-38486 . Similar to the synthetic glucocorticoid (GC) receptor agonist dexamethasone, pharmacological doses of the immunomodulator linomide (quinoline-3-carboxamide) prevent death in all four models of lethal septic shock (LPS + GalN, LPS + RU-38486, SEB + GalN, and SEB + RU-38486) and inhibit the secretion of tumor necrosis factor, one of the major intermediate effector molecules of SEB and LPS toxicity . In this system, cyclosporine A (CsA), although effective in suppressing SEB toxicity, fails to counteract the lethal effect of LPS . This observation, together with the fact that linomide acts in the presence of excess amounts of GC receptor antagonist, indicates that linomide functions in a different way to that of known immunosuppressive agents like CsA and GC. Nurs Clin North Am, 1993 Sep, 28(3), 625 - 37 Methicillin-resistant Staphylococcus aureus . Past, present, and future; Morita MM; The MRSA control program at UCDMC is one of many approaches used in hospitals today . With this less restrictive program, we have been able to maintain control of MRSA for approximately 5 years . By establishing a threshold for investigation at four new nosocomial cases per month, the ICD has been able to maintain a manageable endemic level of MRSA . Figure 2 shows the downward trend of new cases of MRSA per fiscal year . During 1991, however, we experienced clustering of cases in two areas of the hospital along with a major outbreak in our burn unit . This is reflected in the significant rise in the number of new cases for that year . Even with these outbreaks, however, we have been able to re-establish control . Figure 3 represents the number of new MRSA cases per month during fiscal year 1991 . After the peak in April, we have again seen a downward trend, with an average of four new cases per month during the first quarter of fiscal year 1992 . The success of this program is attributed to the cooperation and excellent communication among the hospital departments who are active participants in this control program . It is also owing to the establishment of a very active antibiotic control program that discourages inappropriate use of antimicrobial agents . This team effort is critical to the success of any infection control program . Infection control protocols are constantly changing, even if they are directed toward the same pathogen . Hospital epidemiology provides us with a very systematic approach to controlling the spread of disease . During the process, however, we are constantly collecting new information about the pathogen, allowing us to re-evaluate and, we hope, improve our approach to control . This is one reason for the numerous approaches to control found in the vast array of MRSA literature . MRSA continues to be a challenge of the 1990s . It appears to be an increasing problem in US hospitals, regardless of hospital size . Although its elusive behavior evades the most comprehensive and carefully planned control programs of even the most skilled hospital epidemiologists, hospitals continue to struggle for control of this organism . The alternative to control is probable resistance to vancomycin, which would have devastating public health consequences, particularly as alternative antibiotic therapy is not yet available. Infect Immun, 1993 Sep, 61(9), 3879 - 85 The accessory gene regulator (agr) controls Staphylococcus aureus virulence in a murine arthritis model; Abdelnour A et al.; We have studied the role of the accessory gene regulator (agr) of Staphylococcus aureus as a virulence determinant in the pathogenesis of septic arthritis . At least 15 genes coding for potential virulence factors in Staphylococcus aureus are regulated by a putative multicomponent signal transduction system encoded by the agr/hld locus . agr and hld mutants show a decreased synthesis of extracellular toxins and enzymes, such as alpha-, beta-, and delta-hemolysin, leucocidin, lipase, hyaluronate lyase, and proteases, and at the same time an increased synthesis of coagulase and protein A as compared with the wild-type counterpart . We have used a recently described murine model of S . aureus-induced arthritis to study the virulence of S . aureus 8325-4 and two agr/hld mutants derived from it . Sixty percent of the mice injected with the wild-type strain developed arthritis, whereas agrA and hld mutants displayed joint involvement in only 10 and 30%, respectively . In addition, 40% of the mice inoculated with the wild-type strain displayed an erosive arthropathy; such changes were not detectable at all in mice inoculated with the agrA mutant . Serum levels of interleukin-6, a potent B-cell differentiation factor, were significantly higher (P < 0.001) in the mice inoculated with the wild-type strain than in those inoculated with the agrA mutant counterpart . Overall, our results suggest that the agr system of S . aureus is an important virulence determinant in the induction and progression of septic arthritis in mice. J Pediatr, 1993 Sep, 123(3), 422 - 4 Microbiology of kerions; Honig PJ et al.; Qualitative and semiquantitative bacterial culture specimens were obtained from 44 patients with kerions . Staphylococcus aureus was isolated from the scalp surface overlying the kerion in 29% of patients and from the pus within the kerion in 48% of patients . Gram-negative bacteria were found in the same locations in 11.3% and 18% of patients, respectively . The data indicate that bacteria are frequently cultured from kerions. Eur J Epidemiol, 1993 Sep, 9(5), 559 - 62 Antibiotic susceptibility and phage typing of methicillin-resistant Staphylococcus aureus clinical isolates from blood cultures of 692 patients in 15 Israeli hospitals; Samra Z et al.; Six hundred ninety-two clinical isolates of Staphylococcus aureus were collected from blood cultures of 692 patients in 15 Israeli hospitals over a two year period . Antibiotic sensitivity was tested by the standard disk diffusion technique . Of these isolates, 41.6% were methicillin-resistant (MRSA) . All 288 MRSA isolates were sensitive to vancomycin and pristinomycin; 98.6% were sensitive to fucidine; 97.9% to imipenem; 79% to rifampicin; 63.6% to amikacin; 54.5% to augmentin; 36.4% to clindamycin; 12.6% to ciprofloxacin; 11.9% to cotrimoxazole and ofloxacin; 10.5% to gentamicin; 9.8% to erythromycin; and 8.4% to norfloxacin . Phage typing was determined by using the international set of phages . All the isolates that were sensitive to Group I phages, and 91.8% of those sensitive to Group II phages, were sensitive to methicillin . Of the isolates that were sensitive to Group III phages, 79.2% were methicillin-resistant and 72.4% of the latter were sensitive to phages 75/85 . Of the isolates that were sensitive to Group III and miscellaneous phages, 50.7% were methicillin-resistant and 71% of the latter were sensitive to phages 75/85 as well. Microb Pathog, 1993 Sep, 15(3), 227 - 36 Immunization with fibronectin binding protein from Staphylococcus aureus protects against experimental endocarditis in rats; Schennings T et al.; Rats were immunized with a fusion protein (gal-FnBP) encompassing beta-galactosidase and the domains of fibronectin binding protein from Staphylococcus aureus responsible for binding to fibronectin . Antibodies against gal-FnBP were shown to block the binding of S . aureus to immobilized fibronectin in vitro . Endocarditis in immunized and non-immunized control rats was induced by catheterization via the right carotid artery, resulting in damaged aortic heart valves which became covered by fibrinogen and fibronectin . The catheterized rats were then infected intravenously with 1 x 10(5) cells of S . aureus . The number of bacteria associated with aortic valves was determined 1 1/2 days after the challenge infection and a significant difference in bacterial numbers between immunized and non-immunized groups was then observed (p < 0.05). J Antimicrob Chemother, 1993 Sep, 32(3), 453 - 8 Efficacy of cefoperazone in combination with sulbactam in experimental Staphylococcus aureus endocarditis in rabbits; Chambers HF et al.; The activity of cefoperazone with and without sulbactam was studied in vitro and in vivo against strains of methicillin-resistant and methicillin-susceptible Staphylococcus aureus . Cefoperazone with or without sulbactam was inactive in vitro against the methicillin-resistant strain and was bound by penicillin-binding protein 2a with an IC50 of 190 mg/L (the concentration that reduced radio-labelling with 3H-penicillin by 50%) . Cefoperazone was hydrolysed by beta-lactamase in vitro but sulbactam improved cefoperazone activity in a rabbit model of aortic valve endocarditis caused by a beta-lactamase producing methicillin-susceptible strain. J Antimicrob Chemother, 1993 Sep, 32(3), 431 - 44 Behaviour of beta-lactamase-positive and -negative Staphylococcus aureus isolates in susceptibility tests with piperacillin/tazobactam and other beta-lactam/beta-lactamase inhibitor combinations; Bonfiglio G et al.; beta-Lactamase production protects Staphylococcus aureus against piperacillin and amoxycillin . Tazobactam and clavulanate inhibit the enzyme, but beta-lactamase producers remained substantially less susceptible than non-producers to piperacillin/tazobactam and co-amoxiclav in disc tests . These observations reflected the fact the inhibition zones for beta-lactamase non-producers expanded with time, as bacterial growth was killed by the diffusing antibiotic, whereas those for enzyme producers maintained a constant diameter after they first became discernible, indicating equilibrium between diffusion and destruction of the antibiotic . The antibiotic destruction required for this equilibrium depended on biomass formation, which was apparent from the luxuriant growth on beta-lactamase-producers around the inhibition zone edges, and on beta-lactamase induction, which was demonstrable when S . aureus cultures were exposed to discs containing tazobactam alone . beta-Lactamase-related effects on the activity of inhibitor combinations were apparent also in MIC tests, but only when these were performed with large inocula . Thus, the MICs of piperacillin or amoxycillin with tazobactam or clavulanate were only two- to four-fold higher for beta-lactamase producers than non-producers at low inocula (10(4) cfu), but this differential increased to 6- to 16-fold at high inocula (10(6) cfu) . Inoculum effects were greater for tazobactam combinations than for clavulanate combinations, irrespective of the partner penicillin . These beta-lactamase-related inhibition zone differentials and inoculum effects for the inhibitor combinations resemble previous observations for first-generation cephalosporins, notably cephazolin and cephaloridine . Their significance for the latter compounds has occasioned long debate. J Antimicrob Chemother, 1993 Sep, 32(3), 401 - 11 Antimicrobial susceptibility testing of enterococci: results of a survey conducted by the United Kingdom National External Quality Assessment Scheme for Microbiology; Snell JJ et al.; Six strains of enterococci were distributed to participants in the United Kingdom National External Quality Assessment Scheme for Microbiology with a request that they be tested for susceptibility to ampicillin/penicillin and vancomycin and for high-level resistance to gentamicin . More than 98% of laboratories correctly reported the resistant strains as resistant to penicillin and ampicillin . However, there was a poorer correlation for the susceptible strains, with only 1-6% and 69-83% of laboratories reporting the strains as susceptible to penicillin and ampicillin respectively . Those laboratories which used 5-25 micrograms ampicillin discs or breakpoint methods reported proportionally more results correctly for the susceptible strains than those using 2 micrograms discs . Participants using Escherichia coli or an enterococcus as a control and those not using a control at all were more likely to report ampicillin-susceptible strains correctly than those using Staphylococcus aureus NCTC 6571 (Oxford strain) . For vancomycin-susceptible and high-level vancomycin-resistant strains, 93-96% and 96-99% of reports respectively were correct . For the low-level vancomycin-resistant strains, only 50-54% of reports were correct . Participants using a 5 micrograms vancomycin disc reported proportionally more of the low-level resistant strains correctly than those using 10, 20, 25, 30 or 100 micrograms discs . Participan