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| Pediatr Pulmonol, 1995 Aug, 20(2), 83 - 8 Staphylococcal pneumonia in childhood: will early surgical intervention lower mortality? Joosten KF, Hazelzet JA, Tiddens HA, Hazebroek FW, Dzoljic-Danilovic G, Neijens HJ, de Groot R. Staphylococcus aureus pneumonia (SAP) continues to be a serious bacterial infection which is associated with a high incidence of complications . We retrospectively reviewed the case records of 36 infants and children admitted with SAP to the Sophia Children's Hospital between 1970 and 1992 to analyse changes over time in the clinical presentation, diagnostic work-up, management and complications . Fifteen of these 36 children (42%) were less than 1 year old . Fever (97%) and respiratory distress (83%) were the most common symptoms at the initial presentation . Chest X-ray findings on admission or during hospitalisation included pleural effusion (75%), pneumothorax (47%), and abscess and/or pneumatocele (39%) . Diagnostic and/or therapeutic thoracentesis of pleural fluid was performed in 17 of the 36 patients (47%) . Twenty-one patients (58%) needed chest tube drainage . Twelve had a thoracotomy (33%) . Artificial ventilation was needed in 13 of the patients (36%) . Extrapulmonary complications included convulsions in 6 patients (17%) and osteomyelitis in 2 children (6%) . The mean duration of hospitalization was 36 days . Two of the 36 children died (6%) . The low mortality rate in this study may be the result of the relatively high rate of thoracotomy and of improvements in supportive treatment. J Chemother, 1995 Aug, 7(4), 355 - 62 In vitro postantibiotic effect and postantibiotic leukocyte enhancement of tobramycin; Novelli A et al.; The occurrence of a postantibiotic effect (PAE) and postantibiotic leukocyte enhancement (PALE) of tobramycin, a natural aminoglycoside clinically used since the early 1970s, has been investigated in comparison to gentamicin on recent clinical isolates of Staphylococcus aureus (both methicillin-susceptible and -resistant strains) and of Gram-negative fermenting and non-fermenting rods . A concentration-dependent PAE was observed with both antibiotics, regardless of the bacterial species used, with some variability based on their intrinsic resistance . Tobramycin, at concentrations equal to or higher than the minimum inhibitory concentration (MIC), exhibited a rather long PAE (ranging from 1.9 to 10.9 h) which was often significantly longer than that observed with gentamicin (ranging from 1.0 to 7.5h) . Moreover, pre-exposure to tobramycin led to enhanced polymorphonuclear leukocyte phagocytosis and killing with a 2- to 27-fold increase in activity compared to controls . These results suggest that tobramycin might be conveniently used with once-daily dosing for the treatment of infections due to sensitive pathogens. Clin Infect Dis, 1995 Aug, 21(2), 433 - 4 Sporotrichoid lymphangitis due to Staphylococcus aureus in a diabetic patient; Lieberman AA et al.; We describe a case of sporotrichoid lymphangitis (nodular lymphangitis or lymphocutaneous syndrome) that was an unusual clinical presentation of Staphylococcus aureus infection in a diabetic patient . Common pyogenic bacteria should be considered in the differential diagnosis of sporotrichoid lymphangitis. Clin Infect Dis, 1995 Aug, 21(2), 328 - 32 Familial carriage of methicillin-resistant Staphylococcus aureus and subsequent infection in a premature neonate; Hollis RJ et al.; During routine surveillance of patients in a Neonatal Intensive Care Unit (NICU), an alert infection-control practitioner confirmed the relationship of the index patient (sibling 3) who had a methicillin-resistant Staphylococcus aureus (MRSA) infection to an infant sibling (sibling 2) who had been admitted to the hospital 7 months previously with an MRSA infection . Cultures of nasal specimens obtained from the index patient's parents and two other siblings also yielded MRSA for two of the family members, the mother and sibling 1 . The strains were typed by antibiogram, plasmid analysis, and genomic DNA typing . The isolates from sibling 1, sibling 2, the mother, and one isolate from sibling 3 were found to be identical by all techniques . The other isolates from sibling 3 shared the same genomic type but had no detectable plasmids . These findings suggest that transmission of this strain occurred at least three times within this family and that at least one family member was colonized with the same strain for 7 months or more . Recognition that family members may serve as reservoirs for nosocomial infections with MRSA raises important issues for infection control. J Biochem (Tokyo), 1995 Aug, 118(2), 271 - 7 Stimulation of phagocytosis and phagosome-lysosome fusion by glycosphingolipids from Sphingomonas paucimobilis; Miyazaki Y et al.; Sphingomonas paucimobilis, a Gram-negative opportunistic pathogen, is actively phagocytosed by human peripheral polymorphonuclear leukocytes (PMN) in vitro . However, when live or killed cells were delipidated, the phagocytic rate was clearly decreased . Therefore, we have investigated the physiological role of membrane lipids in phagocytic processes . S . paucimobilis type strain 2395 produces four classes of acidic glycosphingolipids (GL-1, GL-2, GL-3, and GL-4) with the common components of glucuronic acid, 2-hydroxy myristic acid and d18:0 or d21:1 long-chain base . The effect of acidic glycosphingolipids on phagocytosis by PMN using killed Staphylococcus aureus cells coated with glucuronosyl ceramide (GL-1) or ceramide tetrahexoside (GL-4) was also examined . The rate of phagosome-lysosome fusion by PMN was determined by counting acridine orange-stained bacteria under a fluorescence microscope . Both phagocytosis and phagosome-lysosome fusion by PMN of glycosphingolipid-coated bacteria were stimulated markedly in a dose-dependent manner . It was noted that GL-1 or GL-4 stimulated phagosome-lysosome fusion dramatically, but synthetic lipid A did not . Superoxide anion release from PMN was enhanced significantly by the coating with synthetic lipid A at higher concentration, but only slightly with GL-1 or GL-4 . Glucuronic acid was an inhibitor of phagocytosis of GL-1-coated S . aureus by PMN . The effect of acidic glycosphingolipids obtained from mammalian tissue on phagocytosis was also compared with that of bacterial glycosphingolipids . Ganglioside GM3 and sulfatide showed a marked stimulative activity for phagocytosis by PMN, while the neutral glycosphingolipids did not . Thus, bacterial acidic glycosphingolipid and mammalian acidic glycosphingolipid promote phagocytosis and phagosome-lysosome fusion by PMN. Am J Vet Res, 1995 Aug, 56(8), 997 - 1006 Regulation of neutrophil adhesion molecules and shedding of Staphylococcus aureus in milk of cortisol- and dexamethasone-treated cows; Burton JL et al.; The effects of 3 days of glucocorticoid administration on bovine blood neutrophil expression of L-selectin and CD18, and on the health status of mammary glands subclinically infected with Staphylococcus aureus were measured in 9 lactating Holsteins . The experiment was a 3 x 3 Latin square cross-over design, with 3 glucocorticoid treatments switched among groups of 3 cows/treatment during 3 periods . Treatments consisted of a vehicle (control, 10 ml of excipient/cow/d), cortisol (7.5, 15, and 7.5 mg/cow on days 1, 2, and 3, respectively), and dexamethasone (0.04 mg/kg of body weight/cow/d for total daily dosages that ranged from 21.6 to 33.2 mg) . Blood samples for immunostaining and flow cytometric analysis of L-selectin and CD18 and leukograms, as well as foremilk samples for determination of S aureus shedding, somatic cell counts, protein and fat percentages, and daily milk yields were collected repeatedly before, during, and after treatment days . Dexamethasone caused a profound, acute, short-lived down-regulation of L-selectin on neutrophils, which correlated in time to leukocytosis, mature and immature neutrophilias, increased shedding of S aureus in infected glands, and onset of high percentages of fat and protein and decreased milk yields . Dexamethasone also caused profound but delayed down-regulation of neutrophil CD18, which reached nadir simultaneously with reappearance of L-selectin-bearing neutrophils, normalized blood neutrophil counts, markedly high foremilk somatic cell counts and protein percentage, decreased S aureus shedding in milk, and finally, expression of clinical mastitis in some infected quarters . Each of these variables had returned to control (vehicle) values by the ninth (and last) sample collection day . Although cortisol treatment also decreased expression of L-selectin and CD18 on neutrophils, dosages used in this study were not sufficient to alter the number of circulating cells or to convert subclinical mammary gland infections to clinical mastitis . These results suggest that mammary gland health status can be altered by sudden exposure of blood neutrophils to glucocorticoids, because these steroid hormones caused profound down-regulation of the adhesion molecules that direct neutrophil margination and migration through the vascular endothelium . The results also reinforce the potential disease risk of treating infected animals with potent synthetic glucocorticoids, such as dexamethasone. Immunopharmacology, 1995 Aug, 30(2), 147 - 55 Nigella sativa: effect on human lymphocytes and polymorphonuclear leukocyte phagocytic activity; Haq A et al.; The effects of Nigella sativa (N . sativa) seeds and their soluble fractions were studied in vitro on lymphocyte response to different mitogens and on polymorphonuclear leukocyte phagocytic activity . No stimulatory effect of N . sativa was detected on lymphocyte response to phytohemagglutinin, concanavalin-A or pokeweed mitogen . A stimulatory effect of N . sativa was noticed on the lymphocyte response to pooled allogeneic cells . This effect was more pronounced when the low molecular weight (< 10 kDa) fraction was used and varied from one normal individual to another (25% to 825%) . N . sativa enhanced the production of interleukin-3 by human lymphocytes when cultured with pooled allogeneic cells or without any added stimulator . N . sativa did not, however, enhance or suppress interleukin-2 secretion by mitogen activated peripheral blood mononuclear cells . Interestingly, N . sativa increased interleukin-1 beta, suggesting therefore, that it has an effect on macrophages . It also suppressed the leukocyte chemiluminescence activity using phorbol myristate acetate and Zymosan as stimulants . No effect of N . sativa or its fractions was, however, noticed on bacterial phagocytosis or killing when Staphylococcus aureus was used, indicating that the decrease in chemiluminescence activity in the presence of N . sativa is not relevant to the bactericidal activity. J Vet Med Sci, 1995 Aug, 57(4), 765 - 7 Effects of chitosan on experimental abscess with Staphylococcus aureus in dogs; Okamoto Y et al.; An abscess was developed experimentally by a subcutaneous inoculation of Stapylococcus (S.) aureus T-6 with a 4-cm silk suture in dogs . After draining the pus, the abscess was treated with a suspension of finely granulated chitosan (chitosan group), ampicillin (ampicillin group), or saline (control group) (Day 0) . The chitosan group was further divided into 3 subgroups (0.01, 0.1, and 1.0 mg/subgroups) . Similar treatment was repeated after 4 days (Day 4), followed by euthanasia on Day 8 . The wound cavity contraction rate was calculated by measuring the wound cavity diameter by a sound on Days 0, 4 and 8 . The wound cavity contraction rate was significantly higher in the ampicillin, 0.1 mg chitosan, and 1.0 mg chitosan groups than in the 0.01 mg chitosan and control groups on Days 4 and 8 (p < 0.05) . In the 0.1 and 1.0 mg chitosan groups, the abscess healed completely in 6 out of 11 (55%), and 9 out of 10 cases (90%), respectively, by Day 8 . In the ampicillin group, 4 out of 10 cases (40%) healed completely by Day 8 . No healing occurred in the 0.01 mg chitosan and control groups . Histologically, the granulation tissue formed had abundant vascularization in the 0.1 and 1.0 mg chitosan groups on Day 8. Am J Kidney Dis, 1995 Aug, 26(2), 377 - 80 Infection of a subclavian venous stent in a hemodialysis patient; Guest SS et al.; Endovascular stent placement to prevent restenosis after angioplasty is being increasingly employed . A 63-year-old hemodialysis patient with a right forearm gortex graft developed ipsilateral arm edema, and a right subclavian vein stenosis was diagnosed . This vascular stenosis was presumably secondary to previous placement of temporary access catheters . The subclavian vein stenosis was treated with angioplasty, endovascular stenting, and warfarin, which resulted in resolution of the arm edema . Three weeks after stenting, the patient developed fever to 104 degrees F, chills, and right arm and shoulder edema . All blood cultures grew Staphylococcus aureus, and an Indium-labeled white blood cell scan was positive at the sight of the subclavian stent . Infectious disease consultants recommended urgent removal of the infected stent, but the extensive surgery required posed considerable risk of major morbidity . We elected to conservatively treat the patient . With loss of all upper-extremity access sites, the patient was converted to peritoneal dialysis . Despite the patient's ambulatory status, a femoral venous Hickman catheter was placed and tunneled through the abdominal subcutaneous soft tissue . The patient received 9 weeks of antibiotics by the Hickman catheter with an infusion pump, and warfarin was continued . There has been complete clinical resolution of infection and subclavian thrombosis . Endovascular stents are being used more commonly, and this is the first description, to our knowledge, of a stent infection . The stent infection was successfully managed without surgical removal. Epidemiol Infect, 1995 Aug, 115(1), 51 - 60 Carriage of Staphylococcus aureus among 104 healthy persons during a 19-month period; Eriksen NH et al.; The present study was undertaken to investigate the frequency of the nasal carrier rate of Staphylococcus aureus . The investigation was performed on 104 healthy persons . The total number of swabs performed was 1498 and this resulted in isolation of 522 S . aureus strains . All strains have been identified, tested for antibiotic susceptibility, and phage-typed . The carrier-index (number of positive swabs/number of total swabs for each individual person) was compared with different sampling and culturing methods, phage type, age, and resistance to antibiotics . There was statistical difference in carrier rate according to sex (P < 0.05) . Among the 104 persons 15 (14.4%) were persistent carriers, 17 (16.3%) intermittent carriers, 55 (52.9%) occasional carriers and 17 (16.3%) non-carriers . Among intermittent and occasional carriers the phage-type distribution was different from the S . aureus strains isolated from Danish hospitalized patients in 1992, while the persistent carriers had similar phage-type distribution. J Pediatr, 1995 Aug, 127(2), 231 - 7 Humoral immunity in DiGeorge syndrome; Junker AK et al.; OBJECTIVE: To assess humoral immunity after immunization and natural infection in patients with clinical manifestations of the DiGeorge anomalad . DESIGN: Retrospective review of cases . SETTING: Ambulatory immunology clinic of a tertiary care teaching hospital . PATIENTS: The 13 patients had a symptom complex including congenital heart disease, characteristic facies of the DiGeorge anomalad, possible hypocalcemia, and thymic hypoplasia or aplasia . Molecular and cytogenic studies of 12 patients demonstrated that all had 22q11 microdeletions . METHODS: Serial studies included lymphocyte population enumeration by flow cytometry, lymphocyte proliferation assays with the mitogens phytohemagglutinin and pokeweed mitogen and Staphylococcus aureus, and immunoglobulin quantitation . Specific antibody studies included virus neutralization assays for poliovirus antibodies, and enzyme-linked immunosorbent assay for diphtheria, tetanus, measles, rubella, varicella-zoster virus (VZV), and cytomegalovirus (CMV) antibodies . Avidity of rubella, VZV, and CMV antibodies was tested by enzyme-linked immunosorbent assay modified to include a mild protein denaturant in the first wash after incubation with sera . RESULTS: All patients had a CD3+ cell count greater than 0.500 x 10(9)/L and a CD4+ cell count greater than 0.350 x 10(9)/L) . One patient had low proliferation responses to S . aureus, and one to phytohemagglutinin and pokeweed mitogen . Immunoglobulin levels, compared with those in age-related control subjects, were normal except that two patients had transient, borderline low IgG levels and two had elevated IgA levels . Specific antibody tests showed (No . of patients with positive results/No . tested) the following: diphtheria (13/13); tetanus (13/13); poliomyelitis caused by polio virus type 1 (5/9), type 2 (9/9), and type 3 (8/9); measles (11/13); rubella (11/13); and infection with VZV (5/5) and CMV (7/13) . There were no significant differences in antibody avidity results between patients and control subjects for rubella (mean avidity index, 83.5 +/- 8.79 vs 85 +/- 17.6), VZV (81.6 +/- 3.98 vs 65.1 +/- 12.38), or CMV (69.3 +/- 22.31 vs 73.3 +/- 12.46) . CONCLUSIONS: Patients with "partial" DiGeorge anomalad, defined by clinical and immunologic criteria, can be immunized and for the most part can generate good antibody responses. Invest Ophthalmol Vis Sci, 1995 Aug, 36(9), 1828 - 36 Accessory gene regulator controls Staphylococcus aureus virulence in endophthalmitis; Booth MC et al.; PURPOSE . To evaluate the contribution of toxins to the severity of Staphylococcus aureus endophthalmitis . METHODS . Experimental endophthalmitis was established by injecting rabbit eyes with wild type S . aureus ISP479 and the isogenic attenuated strain, ISP546, defective in expression of the global regulator locus agr . agr regulates expression of at least 19 exoproteins that are potentially important in the pathogenesis of endophthalmitis . Infections were evaluated using electroretinography, slit lamp biomicroscopy, and histology . Two concentrations (approximately 10 and 1000 organisms) of bacteria were injected . RESULTS . The agr- strain consistently resulted in slower loss of b-wave response when compared to the wild type strain, irrespective of inoculum size . Clinical signs were less severe among the agr- group at 24 and 48 hours when 10 organisms were injected . However, when the number of bacteria injected was increased to 1000, earlier onset of clinical signs was observed, with both groups showing maximum cell and flare and a white fundal reflex at 48 hours after infection . Histologic examination of eyes enucleated 36 hours after inoculation revealed that the wild type strain induced focal retinal destruction and mild vitritis, whereas eyes infected with the agr- strain remained completely normal . Histologic examination carried out when loss of B-wave response was 100% revealed that retinal changes for both groups could not be distinguished . CONCLUSIONS . These data indicate that toxin production by S . aureus contributes to severity of endophthalmitis by accelerating the rate of onset of retinal damage . Therefore, toxin-targeting therapies instituted early in the course of infection could preserve retinal function. South Med J, 1995 Aug, 88(8), 863 - 5 Primary psoas abscess due to methicillin-resistant Staphylococcus aureus concurrent with septic arthritis of the hip joint; Ash N et al.; We report an unusual case of a primary psoas abscess due to community acquired methicillin-resistant Staphylococcus aureus in an immunocompetent man . The course of the disease was indolent, expressed as septic arthritis of the hip joint . When the diagnosis was made, 3 months after symptoms began, the patient was treated by surgical evacuation of the abscess and appropriate antibiotics . Full recovery and return to his usual activity followed total hip replacement. Biophys Chem, 1995 Aug, 55(3), 273 - 7 Difference in surface properties between Escherichia coli and Staphylococcus aureus as revealed by electrophoretic mobility measurements; Sonohara R et al.; Electrophoretic mobilities of Escherichia coli and Staphylococcus aureus were measured in media of different pH values and ionic strengths at 310 K and the results were analyzed via a new mobility formula which was derived on the assumptions of uniform charge distribution in the cell surface layer of finite thickness and ion-penetrability in the layer . E . coli was shown to have a more negatively charged and less soft surface than that of S . aureus . It is suggested that electrophoretic mobility measurements can be used to detect the difference in surface structure between gram-positive and gram-negative bacteria. Urology, 1995 Aug, 46(2), 246 - 8 Recurrent Staphylococcus aureus renal abscess in a child positive for the human immunodeficiency virus; Brandeis JM et al.; A 10-year-old girl with the human immunodeficiency virus was found to have a Staphylococcus aureus renal abscess with perinephric extension . The abscess was drained first percutaneously and then surgically, and the patient received a 6-week course of intravenous antibiotics . Three months later, the abscess recurred, necessitating a nephrectomy . The extended morbidity and difficulty of eradicating S aureus suggest that, in immunocompromised patients, early aggressive surgical management is indicated. Infect Immun, 1995 Aug, 63(8), 3143 - 50 Quantitative comparison of clumping factor- and coagulase-mediated Staphylococcus aureus adhesion to surface-bound fibrinogen under flow; Dickinson RB et al.; The contributions of clumping factor and coagulase in mediating Staphylococcus aureus adhesion to surface-adsorbed fibrinogen have been quantified by using a new methodology and analysis . The attachment or detachment kinetics of bacteria were directly observed in a radial flow chamber with a well-defined laminar flow field and a spatially varying shear rate and were quantified by recursively scanning the chamber surface and counting cells via automated video microscopy and image analysis with a motorized stage and focus control . Intrinsic rate constants for attachment or detachment were estimated as functions of shear rate for the wild-type Newman strain of S . aureus and for mutants lacking clumping factor, coagulase, or both proteins on surfaces coated with plasma, fibrinogen, or albumin . Clumping factor, but not coagulase, increased the probability of attachment and decreased the probability of detachment of S . aureus on plasma-coated surfaces; however, both clumping factor and, to a lesser extent, coagulase increased the probability of attachment on the purified-fibrinogen-coated surface . All mutants were resistant to detachment on the purified-fibrinogen-coated surface, suggesting the possibility of an additional adhesion mechanism which was independent of coagulase or clumping factor and effective only for fully attached cells . Together, these results suggest that the presence of clumping factor plays the primary role in enhancing adhesion to surfaces with adsorbed fibrinogen, not only by enhancing the probability of cell attachment but also by increasing the strength of the resulting adhesion. Infect Immun, 1995 Aug, 63(8), 2886 - 91 Morphology of defensin-treated Staphylococcus aureus; Shimoda M et al.; Defensins are a family of broad-spectrum antimicrobial peptides found abundantly in the cytoplasmic granules of mammalian neutrophils and Paneth cells of the small intestine . Defensins are known to form ion channels on the membranes of target cells . These channel formations and the cytotoxicity of defensins are intimately linked . We showed the morphological effects of defensins on the cytoplasmic membranes of Staphylococcus aureus by transmission electron microscopy . S . aureus exposed to defensins developed characteristic mesosome-like structures but did not show remarkable changes in cell walls . Defensins induced such structural changes not only at high concentration but also at low concentrations that were not bactericidal . We also showed that increasing the concentration of NaCl in the reaction mixture completely inhibited the occurrence of membranous changes of target cells exposed to defensins . These findings are, to our knowledge, the first report of morphological changes in gram-positive bacteria treated with defensins . Our results indicate that the first effect of defensins in S . aureus is to damage cytoplasmic membranes directly; they also support previous reports that the cell membrane is the principal target of defensins. Infect Immun, 1995 Aug, 63(8), 2826 - 32 A natural mutation of the amino acid residue at position 60 destroys staphylococcal enterotoxin A murine T-cell mitogenicity; Mahana W et al.; A variety of techniques have been used to identify the amino acid residues of bacterial superantigens involved in their interactions with major histocompatibility complex (MHC) class II and T-cell receptor (TCR) . In this study, we isolated a naturally mutated staphylococcal enterotoxin A (SEA) from three different Staphylococcus aureus strains, in which the amino acid at position 60 has been changed from aspartic acid (D) to asparagine (N) . We then studied the influence of this change on the immunological activities of SEA . Our results demonstrated that this mutation does not affect the capacity of SEA to bind MHC class II molecules and consequently activates human monocytes and peripheral blood lymphocytes . In contrast, mutated SEA failed to stimulate the proliferation of murine splenic lymphocytes of two different strains, and when presented by human MHC class II molecules, it also failed to activate murine cell line 3DT, which expresses the SEA-specific TCR V beta element (V beta 1) . These results indicate that this mutation alters the interaction between SEA and murine TCR . The reactivity patterns of the mutated SEA with two specific anti-SEA monoclonal antibodies suggested that the observed effect of the isolated mutation in the murine system might be due to certain conformational changes in the SEA molecule introduced upon changing the D at position 60 to N . Site-directed mutagenesis of the N residue to D or to glycine reconstituted the ability of SEA to stimulate murine splenic lymphocytes . The different effects of this natural mutation at position 60 on the immunological activities of SEA with murine and human cells highlight the relevance of the affinity and avidity in SEA-TCR interactions in the function of different species or may reflect a difference in epitope specificity. Curr Microbiol, 1995 Aug, 31(2), 71 - 6 Transfer of plasmid-borne resistance from a multiply-resistant Staphylococcus aureus isolate, WBG1022; Udo EE et al.; Staphylococcus aureus isolate, WBG1022, was resistant to penicillin, kanamycin, neomycin, streptomycin, chloramphenicol, trimethoprim, cadmium, and ethidium bromide and harbored plasmids of 34.5, 24.5, 4.4, 3.2, and 2.6 kilobases . The plasmids were transferred in mixed-culture transfer and conjugation experiments . No resistance phenotype was associated with the 2.6-kb plasmid . The 3.2-kb and 4.4-kb plasmids encoded chloramphenicol and streptomycin resistance respectively . The 24.5-kb plasmid, pWBG626, encoded joint resistance to penicillin, kanamycin, neomycin, and ethidium bromide . Resistance to trimethoprim and cadmium were chromosomal . The 34.5-kb plasmid, pWBG661, had no resistance phenotype but was found to be conjugative . It also mobilized the 4.4-kb and 24.5-kb plasmids in WBG1022 . Restriction endonuclease analysis of pWBG661 with EcoRI, ClaI, PvuII, and BglII restriction enzymes demonstrated that pWBG661 was identical to two previously isolated S . aureus conjugative plasmids, pWBG620 and pWBG637, that also lack resistance phenotypes. Nippon Kyobu Geka Gakkai Zasshi, 1995 Aug, 43(8), 1166 - 70 {Successful treatment of an infected pseudoaneurysm of the ascending aorta with omental transfer--a case report}; Inoue T et al.; The formation of pseudoaneurysm is an uncommon complication of after coronary artery bypass grafting (CABG) . We report a 66-year-old man in whom an anastomotic pseudoaneurysm of the ascending aorta derived from mediastinitis after repeat CABG . At operation, the pseudoaneurysm was revealed to be involved in the proximal anastomotic site of a saphenous vein graft to the RCA . The aneurysm was resected and the defect was repaired with woven dacron patch under deep hypothermic circulatory arrest . In addition, omental transposition was performed to treat mediastinitis radically . Debris of the pseudoaneurysm grew methicillin-resistant Staphylococcus aureus (MRSA), so vancomycin was administered intravenously for 8 weeks . The postoperative course was uneventful . We considered that omental transfer can be very effective in the management of severe mediastinitis, especially that due to an infected pseudoaneurysm or widespread mediastinitis caused by MRSA. J Antibiot (Tokyo), 1995 Aug, 48(8), 780 - 6 Antibiotic GE37468 A: a new inhibitor of bacterial protein synthesis . I . Isolation and characterization; Stella S et al.; GE37468 A is a new thiazolyl peptide antibiotic obtained by fermentation of Streptomyces sp . strain ATCC 55365 . It inhibits bacterial protein synthesis by acting on elongation factor Tu and is structurally and functionally related to the GE2270 class of EF-Tu inhibitors . It is active in vitro against Gram-positive bacteria and Bacteroides fragilis, and protects mice against Staphylococcus aureus infection. West J Med, 1995 Aug, 163(2), 128 - 32 Staphylococcus aureus septic arthritis in patients on hemodialysis treatment; Slaughter S et al.; We retrospectively reviewed hospital discharge diagnoses of septic arthritis over an 11-year period (1982 through 1992) at 3 medical centers; 11 episodes of septic arthritis were identified in patients on hemodialysis treatment . Of the 11 episodes, 9 were caused by Staphylococcus aureus; in 8 of 9, the blood cultures were positive for the organism and the infection was monoarticular . Concurrent infection of the dialysis access site occurred in 4 cases . Two patients died (22%) . We postulate that repeated skin trauma and contact with health care personnel and facilities result in a high rate of nasal carriage of S aureus and, hence, an increased risk of bacteremia with its attendant complications such as septic arthritis . The use of mupirocin nasal ointment is reported to eradicate or suppress carriage in a high percentage of patients; some studies report that long-term suppressive therapy reduces the frequency of S aureus bacteremia. J Med Assoc Thai, 1995 Aug, 78(8), 393 - 8 Post-traumatic empyema thoracis in blunt chest trauma; Sriussadaporn S et al.; Three out of 42 patients who had isolated blunt chest injury requiring closed tube thoracostomy developed post-traumatic empyema thoracis . All of them were treated by thoracotomy and evacuation of the infected fluid with multiple chest tube drainage . Cultures of the pleural fluid grew Staphylococcus aureus in these 3 patients . Univariate analysis was performed by using Fisher's exact test which revealed the significance of age in association with the development of empyema thoracis . Multivariate analysis was performed by using Logistic Regression . Although no statistical significance was observed, the analysis revealed that the risk of empyema thoracis increased in elderly patients and in patients who had prolonged placement of thoracostomy tube . Intensive pulmonary care in elderly patients who sustained chest injury and early removal of thoracostomy tube is recommended in order to prevent the development of empyema thoracis. J Clin Microbiol, 1995 Aug, 33(8), 2141 - 4 Molecular genotyping of methicillin-resistant Staphylococcus aureus via fluorophore-enhanced repetitive-sequence PCR; Del Vecchio VG et al.; Methicillin resistance in Staphylococcus aureus is a frequent cause of nosocomial and community-acquired infections . Accurate, rapid epidemiologic typing is crucial to the identification of the source and spread of infectious disease and could provide detailed information on the generation of methicillin-resistant S . aureus (MRSA) strains . The high degree of genetic relatedness of MRSA strains has precluded the use of more conventional methods of genetic fingerprinting . A rapid DNA fingerprinting method that exploits PCR amplification from a DNA repeat sequence in MRSA is described . The random chromosomal distribution of this repeat sequence provides an ideal target for detecting DNA fragment patterns specific to individual MRSA strains . Two PCR fingerprinting methods which use an oligonucleotide primer based on a repetitive sequence found in Mycoplasma pneumoniae are presented . The repetitive element sequence-based PCR (rep-PCR) and fluorophore-enhanced rep-PCR (FERP) can identify epidemic strains among background MRSA . The combination of oligonucleotide primers labeled with different fluorescent dyes allowed simultaneous FERP fingerprinting and mecA gene detection . Eight different fingerprint patterns were observed in MRSA strains collected from different sources . These techniques provide a rapid discriminatory means of molecular epidemiologic typing of MRSA involved in nosocomial infections. J Clin Microbiol, 1995 Aug, 33(8), 2032 - 5 Unusually large number of methicillin-resistant Staphylococcus aureus clones in a Portuguese hospital; Couto I et al.; Methicillin-resistant Staphylococcus aureus (MRSA) has been endemic in Hospital de Santa Maria, a 1,300-bed teaching hospital in Lisbon, Portugal, since the mid-1980s with a prevalence of 30% in 1993 . A total of 54 MRSA and 93 methicillin-susceptible S . aureus (MSSA) isolates recovered during the first 3 months of 1993 were analyzed for the particular mecA polymorphs and Tn554 attachment sites (in the case of MRSA) and for pulsed-field gel electrophoretic patterns . While all MRSA isolates shared a very similar multidrug resistance antibiogram, molecular methods allowed the identification of an unusually large number of genetic backgrounds (24 different pulsed-field gel electrophoresis patterns in 54 isolates) and three different mecA polymorphs among the MRSA strains . Similar large variation in the genetic backgrounds of MSSA was observed . The most frequent mecA polymorph (mecA type I) was found in association with three different Tn554 patterns . Among the MRSA strains of Hospital Santa Maria, we found two clonal types previously described in Portugal: one corresponding to the dominant clone in an MRSA outbreak at the pediatric ward of the Lisbon Hospital Dona Estefania and another one identical to the Iberian epidemic clone identified in several Portuguese hospitals and in MRSA outbreaks in Barcelona and Madrid . This suggests that MRSA clones of Hospital de Santa Maria may have been a reservoir for staphylococcal strains over the past decade. J Clin Microbiol, 1995 Aug, 33(8), 2022 - 6 In vivo stability and discriminatory power of methicillin-resistant Staphylococcus aureus typing by restriction endonuclease analysis of plasmid DNA compared with those of other molecular methods; Hartstein AI et al.; We evaluated test discriminatory power and DNA type alterations among methicillin-resistant Staphylococcus aureus strains by testing 199 sequential isolates from 39 patients collected over 30 to 228 days . Isolates were typed by one or three different methods (restriction endonuclease analysis of plasmid DNA {REAP} with or without pulsed-field gel electrophoresis of genomic DNA {PFGE} and immunoblotting {IB}) . REAP was highly discriminatory compared with PFGE and IB . However, the initial isolates from 4 of the 39 patients lacked detectable plasmid DNA and could not be typed by REAP . Typing of individual patient isolates showed that a different REAP type was identified only once every 138 days . Among 25 comparisons, seven sequential isolate pairs demonstrating REAP differences were also different by PFGE and IB . This likely represented the presence of more than one strain . Eighteen other pairs with REAP differences were identical or related to one another by PFGE and IB typing, and 17 of these differences were likely caused by a single genetic alteration within the same strain or clone . The rate of PFGE differences explicable by single genetic alterations among sequential isolates identical by REAP was similar to the overall rate for REAP differences in the whole collection . We conclude that REAP and PFGE typing differences explicable by single genetic alterations are relatively infrequent but not rare . These isolates should be examined by alternative typing systems to further support or refute clonality. Immunology, 1995 Aug, 85(4), 645 - 50 Effects of sodium fusidate in animal models of insulin-dependent diabetes mellitus and septic shock; Nicoletti F et al.; We have evaluated the effects of the novel immunosuppressant sodium fusidate (fusidin) in the non-obese diabetic (NOD) mouse and in D-galactosamine (D-Gal)-presensitized BALB/c mice challenged with the bacterial superantigen, Staphylococcus aureus enterotoxin B (SEB) or with the endotoxin, Escherichia coli lipopolysaccharide (LPS) . The NOD mouse model has clinical and histoimmunological features similar to those of human insulin-dependent diabetes mellitus (IDDM) . The SEB- and LPS-treated BALB/c mouse models exhibit pathogenic similarities with human septic shock conditions . In the NOD mouse, fusidin suppressed the spontaneous development of insulitis (mean inhibition 73%) and hyperglycaemia (IDDM incidence 25% versus 0%) when administered at 40 mg/kg five times weekly for 8 consecutive weeks from the fourth week of age; concurrently treated animals exhibited reduced percentages of splenic T lymphocytes . This anti-diabetogenic effect was confirmed in the accelerated model of diabetes induced in the NOD mouse with cyclophosphamide (CY) (IDDM incidence 55% versus 21-6% using dosages of fusidin from 40 to 80 mg/kg five times weekly); protection from IDDM development was achieved even when the drug (80 mg/kg/day) was first administered 7 days after CY challenge . In contrast, fusidin did not reverse hyperglycaemia when administered to CY-treated animals within 3 days of IDDM development . In the two models of septic shock, prophylactic treatment with fusidin, 80 mg/kg given three times for 2 days prior to D-Gal/SEB or D-Gal/LPS challenge, drastically reduced the lethality compared with D-Gal/buffer-treated mice . This effect may depend on the inhibitory action of fusidin on the secretion of cytokines such as interferon-gamma and tumour necrosis factor-alpha, the serum levels of which were greatly diminished in the fusidin-treated mice (mean inhibition 50-90%) . These results demonstrate that fusidin may have a role in the treatment of cell-mediated autoimmune diseases and cytokine-mediated infectious diseases in humans. Br J Dermatol, 1995 Aug, 133(2), 326 - 7 Recurrent chancriform pyoderma: report of a case with tongue lesions . Is Staphylococcus aureus implicated? Holmes SC, Thomson J. We report a case of recurrent chancriform pyoderma . Staphylococcus aureus was isolated from only one of seven lesions . Immunological responses to S . aureus, as measured by peripheral neutrophil function, were normal . We question the notion that chancriform pyoderma represents an abnormal immune response to a primary staphylococcal infection . To our knowledge, this is only the third reported patient with lesions affecting the tongue. Br J Dermatol, 1995 Aug, 133(2), 319 - 21 A case of Behçet's disease aggravated by gingival infection with methicillin-resistant Staphylococcus aureus; Suga Y et al.; We report a case of Behcet's disease aggravated by gingivitis and carious teeth infected with methicillin-resistant Staphylococcus aureus . Recurrent severe ulcers in the mouth, and on the genitalia and legs were closely linked with the infection, and dramatically improved after extraction of the carious teeth and administration of systemic vancomycin hydrochloride. Burns, 1995 Aug, 21(5), 387 - 8 Pelvic abscess induced by a methicillin-resistant Staphylococcus aureus from haematogenous spread via the CVP line in a burn patient; Lin TW et al.; A 38-year-old female patients was found accidentally to have a positive culture of MRSA from a routine CVP catheter tip culture 1 week after she had complete wound closure . She was recovering from a partial skin thickness burn covering 42 per cent TBSA on the trunk and extremities . Fever and hip pain developed abruptly 1 week later when she was ready for discharge from hospital . Magnetic resonance imaging (MRI) of the pelvis disclosed an intramuscular abscess . Open drainage was performed and pus culture yielded a MRSA with the same sensitivity profile as the previous CVP tip culture . Vancomycin 500 mg every 6 h was used for 3 weeks until the drain culture disclosed a negative result, and a follow-up MRI indicated a loss of the abscess space . Follow-up at an outpatient clinic 3 months later showed that the patient remained symptom free . In this patient haematogenous dissemination was the most likely route of pelvic abscess formation . It should be remembered that MRSA infection is not always only a local problem, especially in the immunocompromised condition of burn injury. J Bacteriol, 1995 Aug, 177(15), 4437 - 41 CadC, the transcriptional regulatory protein of the cadmium resistance system of Staphylococcus aureus plasmid pI258; Endo G et al.; The CadC protein from the cadA cadmium resistance operon of Staphylococcus aureus plasmid pI258 regulates transcription of this system in vitro . The CadC protein was overproduced in Escherichia coli cells and partially purified . Gel shift assays of the proposed cadA operator/promoter region DNA showed specific association with the CadC protein . Control arsenic resistance operator/promoter DNA from the same plasmid was not shifted by the CadC protein . Cd2+, Bi3+, and Pb2+ caused the release of CadC from DNA in gel retardation assays . DNase I footprinting measurements showed that the CadC protein specifically associated with and protected a region of operator/promoter DNA from nucleotide positions -7 to +14 relative to the start point of mRNA synthesis . Runoff transcription assays with the operator/promoter region of DNA (plus the first 69 nucleotides of the cadC gene) and purified E . coli RNA polymerase gave an mRNA product of the predicted size . Added CadC protein inhibited transcription in vitro. J Infect Dis, 1995 Aug, 172(2), 497 - 505 Decreased activation of the respiratory burst in neutrophils from AIDS patients with previous Pneumocystis carinii pneumonia; Laursen AL et al.; Neutrophils from human immunodeficiency virus (HIV)-negative blood donors, asymptomatic HIV-positive patients, AIDS patients with previous Pneumocystis carinii pneumonia (PCP), and AIDS patients without previous PCP were compared for their ability to activate the respiratory burst, measured as luminol-amplified chemiluminescence . P . carinii, Staphylococcus aureus, phorbol-12-myristate-13-acetate, and FMLP were used to stimulate the neutrophils . When stimulated with P . carinii, neutrophils from PCP patients had a significantly lower response than the other groups, whereas no difference was found when S aureus was used . A somewhat but not significantly lower response to P . carinii was also seen in non-PCP patients compared with HIV-negative donors . Priming of the neutrophils with recombinant granulocyte colony-stimulating factor (G-CSF) or recombinant human granulocyte-macrophage (GM)-CSF corrected this defect . A similar effect of these cytokines was seen on phagocytosis, whereas the chemiluminescence in unprimed cells did not correlate with phagocytosis. J Infect Dis, 1995 Aug, 172(2), 410 - 9 Growth of Staphylococcus aureus with nafcillin in vitro induces alpha-toxin production and increases the lethal activity of sterile broth filtrates in a murine model; Kernodle DS et al.; The morbidity and mortality of Staphylococcus aureus infections remain high despite antibiotic therapy . To investigate further the observation that penicillins increase the hemolytic activity of staphylococcal cultures, 37 strains were grown in broth with and without subinhibitory nafcillin . Nafcillin stimulated hemolytic activity in nafcillin-susceptible and -resistant isolates . Sterile broth filtrates of nafcillin-associated cultures injected intraperitoneally in mice were more rapidly lethal than filtrates of the same strain grown without nafcillin . Lethality was neutralized by anti-alpha-toxin antisera . DNA-RNA hybridization revealed a nafcillin-associated increase in alpha-toxin mRNA during the postexponential growth phase after the activation of agr . Isolates grown in slightly inhibitory nafcillin concentrations had more alpha-toxin mRNA than did nafcillin-free cultures, whereas agr RNAIII levels were comparable . This suggests that nafcillin-induced alpha-toxin production is not entirely attributable to agr . A supplemental regulatory mechanism may be involved. Med Microbiol Immunol (Berl), 1995 Aug, 184(2), 53 - 61 The epidermolytic (exfoliative) toxins of Staphylococcus aureus; Bailey CJ et al.; Two epidermolytic toxins, produced by different strains of Staphylococcus aureus, split human skin at a site in the upper epidermis . Clinical effects are most common in infants, but adults are susceptible . Epidermolysis may also be observed in the mouse, in vivo and in vitro, and in a few other mammals . Recent in vitro experiments have demonstrated an inhibition by chelators and point to metal-ion, possibly Ca2+, involvement . The epidermolysis effect is insensitive to a wide range of other metabolic inhibitors . The toxin amino acid sequences are similar to that of staphylococcal proteinase, and new experiments by chemical modification and site-directed mutagenesis have shown that toxicity depends on 'active serine' residues of a catalytic triad similar to that found in serine proteases . Furthermore the toxins possess esterolytic activity, also dependent on the 'active serine' sites . However, the toxins have low or undetectable activity towards a range of peptide or protein substrates . In histological and related studies, the toxins bound selectively to an intracellular skin protein, profilaggrin, but there was no evidence that the toxin can enter intact epidermal cells . Therefore, although the circumstantial evidence that the toxins act by proteolysis is convincing, a specific skin proteolytic substrate for the toxin has not been identified. Antimicrob Agents Chemother, 1995 Aug, 39(8), 1752 - 5 Lipid-based slow-release formulation of amikacin sulfate reduces foreign body-associated infections in mice; Roehrborn AA et al.; Treatment and prophylaxis of uncomplicated infections with standard systemic antibiotics are usually successful . However, standard systemic antibiotic therapy alone is frequently unsatisfactory in certain circumstances, such as the presence of a foreign body (FB), necrotic tissue, overwhelming bacterial inoculum, or poor vascular supply to the involved tissues . We have developed a lipid-based sustained release formulation of amikacin sulfate (DepoFoam encapsulated amikacin sulfate {DEAS}) as a biodegradable, locally injectable antibiotic for such circumstances . The encapsulated drug is released over 7 to 10 days . We tested the efficacy of this formulation in an FB infection model in which Teflon tubes (length, 1 cm; outside diameter, 1.6 mm) were implanted into the subcutaneous tissue in mice and the local site was inoculated with 0.87 x 10(7) CFU of Staphylococcus aureus 3 days later . Inoculation was followed by either no treatment or a local injection of DEAS, free amikacin sulfate, non-drug-containing DepoFoam, or systemic free amikacin sulfate . All drug applications contained 1 mg of amikacin . One group was implanted with the FB and left unchallenged with bacteria and untreated as a sterile control group . All animals were sacrificed 10 days following FB implantation . FBs were retrieved from tissues by an aseptic technique and incubated in liquid culture media for 7 days . Local wound tissue was excised and processed to determine the number of CFU per gram of tissue . Treatment with local or systemic free amikacin had no effect on the number of infected FBs or on the log CFU in wound tissue compared with the untreated or non-drug-containing DepoFoam group . Compared with local free amikacin therapy, the number of infected FBs was reduced from 86 to 25% (P=0.02) following treatment with DEAS, and log CFU per gram of tissue was significantly decreased from 4.8 +/- 0.9 to 1.3 +/- 0.6 (P<0.0005) . DEAS may have clinical utility as locally injected antibiotic in certain infections. Antimicrob Agents Chemother, 1995 Aug, 39(8), 1748 - 51 Influence of aspirin on development and treatment of experimental Staphylococcus aureus endocarditis; Nicolau DP et al.; Previously, we have shown that a 5-mg/kg of body weight daily dose of aspirin (ASA) caused reductions in the bacterial densities and weights of aortic vegetations in a rabbit model of Staphylococcus aureus endocarditis . We sought to determine (i) whether ASA dosage influences the development of vegetations and (ii) whether ASA given with antimicrobial therapy improves the treatment outcome of infective endocarditis . To study the influence of ASA dosage, animals received either no ASA (control) or oral doses of 2.5, 10, 20, and 50 mg/kg daily . The 2.5- and 10-mg/kg groups had statistically significant reductions in vegetation weight compared with untreated controls . The 10-mg/kg dose also resulted in a significant decrease in bacterial densities compared with those of the controls . Although reductions in weight and bacterial density were observed in other ASA-treated groups, these did not achieve statistical significance . To study the influence of ASA and antimicrobial therapy, the animals received either vancomycin alone or vancomycin with ASA . When ASA was given prior to and during antimicrobial therapy, a significant reduction in vegetation weight was observed . Additionally, the rate of sterilization was directly proportional to this observed reduction in weight . ASA's impact on the reduction of both the bacterial density and the weight of aortic vegetations is a dose-dependent phenomenon . When given with antimicrobial therapy, ASA not only reduces vegetation weight but also improves the rate of sterilization . This study provides additional data regarding the role of ASA in the treatment of endocarditis. Antimicrob Agents Chemother, 1995 Aug, 39(8), 1655 - 60 Comparison of sparfloxacin, temafloxacin, and ciprofloxacin for prophylaxis and treatment of experimental foreign-body infection by methicillin-resistant Staphylococcus aureus; Cagni A et al.; The prophylactic and therapeutic activities of three broad-spectrum fluoroquinolones were evaluated in two different experimental models of foreign-body infections caused by methicillin-resistant Staphylococcus aureus (MRSA) susceptible to quinolones . In a guinea pig model of prophylaxis, subcutaneously implanted tissue cages were infected at a > 90% rate by 10(2) CFU of MRSA in control animals . A single dose of 50 mg of ciprofloxacin per kg of body weight administered intraperitoneally 3 h before bacterial challenge was less effective than an equivalent regimen of either sparfloxacin or temafloxacin in decreasing the rate of experimental infection in tissue cages challenged with increasing inocula of MRSA . In a rat model evaluating the therapy of chronic tissue cage infection caused by MRSA, the efficacy of a 7-day high-dose (50-mg/kg twice-daily) regimen of sparfloxacin, temafloxacin, or ciprofloxacin was compared to that of vancomycin (50 mg/kg twice daily) . Active levels of sparfloxacin, temfloxacin, or ciprofloxacin were continuously present in tissue cage fluid during therapy, exceeding their MBCs for MRSA by 6- to 20-fold . Either temafloxacin, sparfloxacin, or vancomycin was significantly (P < 0.01) more active than ciprofloxacin in decreasing the viable counts of MRSA in tissue cage fluids . The different activities of ciprofloxacin compared with those of the other two quinolones against chronic tissue cage infections caused by MRSA did not involve the selective emergence of quinolone-resistant mutants . Temafloxacin and ciprofloxacin, which showed the most prominent differences in their in vivo activities, however, exhibited similar bactericidal properties and pharmacokinetic parameters in the rat model . In conclusion, both temafloxacin and sparfloxacin were significantly more active than ciprofloxacin for the prophylaxis or treatment of experimental foreign-body infections caused by a quinolone-susceptible strain of MRSA. Photochem Photobiol, 1995 Aug, 62(2), 342 - 7 Structure-activity relationship of porphines for photoinactivation of bacteria; Nitzan Y et al.; The antibacterial photodynamic effects of uncharged (o-tetrahydroxyphenyl porphine {THPP}, m-THPP and p-THPP), cationic (5,10,15,20-tetra{4-N-methylpyridyl}porphine {TMPyP}) and anionic (5,10,15,20-tetra{4-sulfonatophenyl porphine} {TPPS4}) porphines on Staphylococcus aureus and Escherichia coli bacteria inactivation were examined . The results show that uncharged porphines provoked antibacterial photodynamic activity on S . aureus, and also on E . coli in the presence of the membrane-disorganizing peptide polymixin B nonapeptide (PMNP) . The TMPyP compound was highly photoactive toward gram-positive bacteria but only marginally effective on gram-negative cells, whereas TPPS4 showed no activity on either gram-positive or gram-negative bacteria . The photoactivity of TMPyP is due to the electrostatic attraction between the positively charged sensitizer molecule and the negatively charged membrane of the gram-positive target cells . For TPPS4, the inactivity toward gram-positive bacteria is due to electrostatic repulsion between the charged sensitizer molecule and the cell membrane . For gram-negative bacteria, the inactivity is conceivably due to preferential (electrostatic) binding to the positively charged PMNP, which is an adjuvant for membrane disorganization, but has no effect on cell viability . For hydrophobic sensitizers, the photoactivity depends on the state of aggregation . The extent of deaggregation of the different THPP isomers was determined by fluorescence measurements of bound sensitizers and could be positively correlated with their photoinactivation capacity . We conclude that the structure-activity relationships of these porphines are affected by their net charge and by aggregation. Ugeskr Laeger, 1995 Jul 24, 157(30), 4249 - 50 {Preclinical hereditary hemochromatosis--is there an indication for preventive screening?}; Gronbaek KE et al.; A 54 year-old previously healthy woman was admitted with staphylococcus aureus septicaemia . The patient had been treated with oral iron supplementation for two years due to fatigue . In the evaluation of postinfectious anaemia, serum transferrin saturation and serum ferritin were found persistently elevated with values of 74% and 950 micrograms/1, respectively . Hereditary haemochromatosis was suspected even though there was no history of liver disease or diabetes mellitus in the family . A bone marrow biopsy showed a normal content of haemosiderin iron . The liver biopsy revealed haemosiderosis, mainly located to the periportal hepatocytes, and fibrosis in the portal tracts . The HLA-type was A3, B7, B37 . Over a period of ten months, a total of 3.9 g of iron was removed by venesection while S-ferritin declined to 31 micrograms/l . A sister to the proband had an identical HLA type, but normal iron status markers, either indicating heterozygosity or homozygosity with lack of penetrance . In preclinical hereditary haemochromatosis, early diagnosis and treatment is essential in order to prevent organ damage and to improve prognosis . Prophylactic screening is recommended . The identification of one homozygous subject in a Danish year-cohort of 60.000 persons costs approximately 40.000 Danish kroner (7.000 US+). J Immunol, 1995 Jul 15, 155(2), 776 - 84 Role of protein kinase C isozymes in Fc gamma receptor-mediated intracellular killing of Staphylococcus aureus by human monocytes; Zheng L et al.; Intracellular killing of Staphylococcus aureus by human monocytes after cross-linking Fc gamma R is known to be a phospholipase C (PLC)-dependent process . Activation of PLC leads to the formation of second messengers that synergistically activate protein kinase C (PKC) . The aim of this study was to obtain more insight into the role of PKC in Fc gamma R-mediated killing process . PKC inhibitors H-7 and staurosporine markedly suppressed the killing of S . aureus by monocytes stimulated by cross-linking Fc gamma RI or -II . Cross-linking Fc gamma R caused a transient increase in PKC activity in the membranes of monocytes, as measured by Ca2+/phospholipid-dependent phosphorylation of histone . Western blot analysis revealed that cross-linking Fc gamma R stimulated a transient increase in PKC-beta in the membranes of monocytes with kinetics that correlated closely with the translocation of PKC activity . Cross-linking Fc gamma R on monocytes also stimulated the translocation of PKC-epsilon but not PKC-alpha . PMA and 1-oleoyl-2-acetylglycerol (OAG), which caused translocation of PKC-alpha, -beta, and -epsilon, did not stimulate the killing process . Incubation with these PKC activators for 10 min rendered monocytes unresponsive to stimulation of killing of S . aureus via Fc gamma R . It could be that activation of certain PKC isozymes, probably PKC-alpha and -epsilon, by these activators causes feedback inhibition of PLC and, consequently, the killing in monocytes, because PMA blocks the Fc gamma R-mediated intracellular inositol(1,4,5)P3 formation and PKC translocation . Together, our results indicate that PKC isozymes play an important role in both stimulation and inhibition of the Fc gamma R-mediated intracellular killing of bacteria by monocytes. J Med Chem, 1995 Jul 7, 38(14), 2531 - 40 Synthesis and antibacterial activity of some novel 1-substituted 1,4-dihydro-4-oxo-7-pyridinyl-3-quinolinecarboxylic acids . Potent antistaphylococcal agents; Reuman M et al.; The palladium-catalyzed coupling of 3- and 4-(trialkylstannyl)pyridines with 7-bromo or 7-chloro 1-substituted 1,4-dihydro-4-oxo-3-quinolinecarboxylates has provided access to the corresponding 1-substituted 1,4-dihydro-4-oxo-7-pyridinyl-3-quinolinecarboxylic acids . The antibacterial activity of these derivatives was studied with the finding that the optimal 1- and 7-position substituents for Gram positive activity are cyclopropyl and 4-(2,6-dimethylpyridinyl), respectively . We find that for the fluorine-substituted derivatives studied, the position of the fluorine on the quinolone nucleus or the number of fluorine atoms does not seem to be important for good Gram positive activity . For 1-cyclopropyl 7-(2,6-dimethyl-4-pyridinyl) derivatives, the 6-fluoro 4a, 8-fluoro 10d, 6,8-difluoro 10b, and 5,6,8-trifluoro 8, all provided equal antibacterial activity against Staphylococcus aureus ATCC 29213 . There is also a correlation between the substitution on the 7-(4-pyridinyl) group and the Gram positive activity, particularly for S . aureus, clearly indicating that the 2,6-dimethylpyridinyl group is optimal . The MIC50 value for the most potent agents in this study against S . aureus ATCC 29213 is 0.008 microgram/mL . By comparison, ciprofloxacin and aminopyrrolidine 28 gave values of 0.25 and 0.015 microgram/mL, respectively, against this organism. J Ethnopharmacol, 1995 Jul 7, 47(2), 97 - 100 Antibacterial activity and phytochemical analysis of Vochysia divergens (Vochysiaceae); Hess SC et al.; Vochysia divergens Pohl (Vochysiaceae) is a tree commonly found in wet soils of 'Pantanal' of Mato Grosso, Brazil, and used in folk medicine against infections and asthma . We have studied different extracts and some isolated compounds from this plant for antibacterial activity . From the extracts of the stem bark beta-sitosterol, betulinic acid and sericic acid were isolated . The minimal inhibitory concentration (MIC) for Staphylococcus aureus were: ethanolic extract (MIC = 1.5 mg/ml); ethyl acetate extract (MIC = 2.0 mg/ml); and sericic acid (MIC = 1.0 mg/ml) . Escherichia coli was resistant until 5 mg/ml. Biochem Biophys Res Commun, 1995 Jul 6, 212(1), 249 - 54 Isolation and characterization of novel antimicrobial peptides, rugosins A, B and C, from the skin of the frog, Rana rugosa; Suzuki S et al.; Three antimicrobial peptides were isolated from the skin of Rana rugosa . The major component, designated rugosin A, consisted of 33 amino acid residues and had structural homology (45%) with brevinin-2 of Rana porosa brevipoda . This peptide strongly inhibited the growth of gram-positive bacteria (e.g . Staphylococcus aureus 209P) . The second peptide (rugosin B), a minor component, also had 33 amino acid residues, but was less homologous (33%) with brevinin-2 . This peptide exhibited a striking antimicrobial activity against both gram-negative (e.g., Escherichia coli NIHJ) and gram-positive bacterial species . The third one, named rugosin C, composed of 37 amino acid residues, exhibited an antimicrobial activity against gram-positive bacteria . All three peptides had an intramolecular disulfide bond at the C-terminus. Biochim Biophys Acta, 1995 Jul 3, 1250(1), 110 - 6 Isolation and characterisation of a vitronectin-binding surface protein from Staphylococcus aureus; Liang OD et al.; In a previous study we demonstrated that cells of Staphylococcus aureus strain V8 bind 125I-labelled vitronectin in a receptor-ligand type of interaction, and a protein having a molecular mass of 60 kDa was identified as a putative high-affinity staphylococcal vitronectin-binding protein (Liang, O.D . et al . (1993) Biochim . Biophys . Acta 1225, 57-63) . In the present communication we report on the isolation and preliminary characterisation of the 60 kDa vitronectin-binding protein . The bacterial cell surface proteins were released by stirring bacteria with 1 M LiCl at 37 degrees C for 2 h and separated on an FPLC Mono-Q column with a gradient of 0-0.5 M NaCl in 20 mM Tris buffer at pH 9.0 . Fractions containing vitronectin-binding activity, assayed on microtiter plates with immobilised human vitronectin, were collected and SDS-PAGE analysis showed the content to be a single protein band at the 60 kDa position . In Western blot experiments the protein transblotted onto nitrocellulose membranes could bind soluble vitronectin . Its amino-terminal amino acid sequences showed a striking similarity with those of a 60 kDa heparan sulfate-binding protein from the same staphylococcal strain (Liang, O.D . et al . (1992) Infect . Immun . 60, 899-906), suggesting that they are identical molecules . This was supported by ligand blotting experiments where both vitronectin and heparan sulfate were shown to bind to the same protein band in parallel strips. Rev Latinoam Microbiol, 1995 Jul-Sep, 37(3), 245 - 55 {Comparison of detection methods for the Staphylococcus aureus capsule}; Odierno L et al.; Eleven methods for capsule detection of Staphylococcus aureus were compared . The most suitable of them were transmission electron microscopy, determination of the presence of clumping factor, determination of colonial morphology in serum-soft agar, estimation of cell volume and staining with safranine . The determination of clumping factor is a fast and effective method for presumptive diagnosis of capsulated strains, but need to be confirmed by another method . The cell volume estimation is useful for determination of capsule production in liquid cultures, while staining with safranine is suitable for genetic studies of capsule production . The other methods analyzed in this work (Indian ink staining, use of anticapsular antisera, determination of virulence for mice, lisostaphin susceptibility, resistance to phages and resistance to phagocytosis) were laborious, too slow, or need components and/or equipment not available in all laboratories . In addition, two methods of induction of capsule production were assayed, one in vitro by several passages in broth with 10% bovine serum and the other in vitro by intraperitoneal inoculation in mice . Both methods induced capsule production. Rev Latinoam Microbiol, 1995 Jul-Sep, 37(3), 237 - 44 {Comparative study of 3 heterologous expression systems for obtaining recombinant Bacillus thuringiensis delta-endotoxins}; Vazquez R et al.; cryIA(b) and cryIA(c) genes encoding active fragments of Bacillus thuringiensis delta-endotoxins were cloned downstream of the pR and pT7 promoters from the lambda and T7 bacteriophages, respectively . cryIA(b) gene was also fused with the gene encoding protein A from Staphylococcus aureus cloned under the control of the pR promoter . There were no remarkable differences in the expression levels of the cloned genes in E . coli, but the Western blot analysis allowed distinct protein quality for the three expression systems . We conclude that the best expression model for the production of delta-endotoxins toxic fragments in E . coli is the one based on lambda pR promoter. J Chemother, 1995 Jul, 7 Suppl 3, 99 - 103 The epidemiology of methicillin-resistant Staphylococcus aureus colonisation and infection; Doebbeling BN; Methicillin-resistant Staphylococcus aureus (MRSA) is an increasingly common nosocomial pathogen in health care facilities throughout the world . Overall, approximately two-thirds of nosocomial cases and outbreaks have occurred in critical care units . Major risk factors for colonisation and infection in nursing homes include age, underlying conditions, nasal colonisation and the presence of indwelling devices such as catheters, tracheostomies and nasogastric tubes . In general, patients with MRSA infections in an acute care facility are more likely to have had a prolonged hospital stay, to have received prior antibiotics and to have severe underlying disease, than patients infected with methicillin-susceptible S . aureus . Risk factors for MRSA bacteraemia include: a higher frequency of severe underlying disease, poorer underlying prognosis, prior antibiotic therapy, prolonged hospitalisation, intravascular catheterisation, and intensive care unit location . Risk factors for developing MRSA postoperative wound infections include: prior antimicrobial therapy, prolonged hospitalisation and severity of underlying disease . Little data are available to identify specific risk factors for colonisation or infection of burn wounds by MRSA. J Chemother, 1995 Jul, 7 Suppl 3, 93 - 8 Aspects of the epidemiology of MRSA in Europe; Cookson B; The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) varies in different European countries, in different cities in the same country or even within a city or a hospital . It is thought that the overall incidence of MRSA is higher than ever before, although the reasons for this are unclear . MRSA typing can identify epidemic MRSA in many countries . The spread of a closely related clone is Northern Europe is a cause for concern, as it is resistant to many agents . In the UK, control measures are threatened by changing patterns of health care and patient demography . Resistance to mupirocin, a valuable agent in the control of MRSA, has emerged in many hospitals in the UK, thus the agent must be used judiciously. J Chemother, 1995 Jul, 7 Suppl 3, 87 - 92 Genetic aspects of methicillin resistance in Staphylococcus aureus and methods used for its detection in clinical laboratories in the United States; Baron EJ; The mecA gene in methicillin-resistant Staphylococcus aureus (MRSA) directs production of a novel penicillin-binding protein (PBP 2A), an enzyme active in cell wall synthesis . MecA, alone, however, does not determine the degree of resistance expressed by strains of MRSA . Differential resistance or variations in other genes that participate in cell wall synthesis, such as laboratory mutant fem genes, may account, in part, for the heterogeneity of methicillin resistance expression in MRSA . The exact mechanisms of methicillin resistance expression in clinical isolates remain to be elucidated . Laboratories use selective agars containing oxacillin and turbidity pattern recognition programs in automated instruments to identify MRSA, although not all mecA-containing strains are detected . Until a rapid and inexpensive DNA probe assay is widely available, newer test methods such as the E test (AB Biodisk), oxidation-reduction indicators in MIC trays (Alamar), and the rapid fluorescent BBl . Crystal system seem promising. J Chemother, 1995 Jul, 7 Suppl 3, 81 - 5 Strategies for controlling methicillin-resistant Staphylococcus aureus in hospitals; Boyce JM; In areas where the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) is very low, aggressive strategies, which appear to have been effective, such as those used in the Netherlands and western Australia, may be feasible . In hospitals where MRSA is epidemic or highly endemic, less rigorous strategies are appropriate . However, which isolation techniques and barrier precautions are optimal is controversial . In addition, there is no consensus regarding the epidemiological importance of environmental contamination . Rapid detection of MRSA, prompt implementation of barrier precautions and prospective surveillance are essential components of a successful control programme . Eradicating nasal carriage of MRSA among patients and personnel can be useful during epidemics, but the cost-effectiveness of using this approach in hospitals where the prevalence of MRSA is low is unknown . Additional studies of this issue need to include surveillance for mupirocin-resistant strains. J Chemother, 1995 Jul, 7 Suppl 3, 67 - 70 Staphylococcus aureus infections during peritoneal dialysis; Coles GA; Peritoneal dialysis is in widespread use for the treatment of chronic renal failure . Infection is still one of the major complications and can include peritonitis and pericannular problems . The rate of peritonitis is currently 0.5 episodes per patient year with disconnect systems, and there are about 0.4 exit-site infections (ESIs) per patient year . ESI is associated with a high rate of catheter removal and replacement . Staphylococcus aureus is a common cause of peritonitis and accounts for more than half of all ESIs . Nasal carriage of S . aureus is associated with a much higher rate of ESI . Treatment of ESIs is unsatisfactory . The type of exit-site care, however, does influence the rate of infection and prophylaxis with oral rifampicin and local or nasal mupirocin has been claimed to reduce ESIs . A large multicentre double-blind trial of nasal mupirocin has just been completed and preliminary results show a reduction in the incidence of S . aureus-induced ESI . The cost benefits of such a regimen are being evaluated. J Chemother, 1995 Jul, 7 Suppl 3, 49 - 53 Staphylococcus aureus infections in haemodialysis patients: pathophysiology and use of nasal mupirocin for prevention; Boelaert JR et al.; Staphylococcus aureus is the most frequently (42%) isolated micro-organism during bacteraemic episodes in haemodialysis patients . Nasal carriage of S . aureus is of major importance in determining the risk of subsequent infections . Indeed, nasal carriage of S . aureus is highly prevalent in uraemic patients from the onset of maintenance dialysis therapy . The strains isolated simultaneously from the nares and the hands are usually the same . Likewise, infecting S . aureus strains and those isolated from nasal surveillance cultures obtained in the same patient are usually similar . S . aureus infections in haemodialysis patients are thus mostly to be considered as auto-infections . The nares are therefore an elective site for the prevention of S . aureus infections in haemodialysis patients . This has been demonstrated with oral rifampin, and more recently with nasal mupirocin, which is highly effective . Long-term application of nasal mupirocin (e.g . once per week) is cost-effective and is only rarely associated with the emergence of mupirocin-resistance in S . aureus. J Chemother, 1995 Jul, 7 Suppl 3, 37 - 43 Identifying high risk patients for Staphylococcus aureus infections: skin and soft tissue infections; Trilla A et al.; Staphylococcus aureus is the causative organism for many skin and soft tissue (SST) infections . Some SST infections have severe systemic complications, such as bacteraemia and sepsis . S . aureus is the cause of 75% of primary pyodermas . Pre-existing conditions, like tissue injury (ulcers, wounds) or tissue inflammation (exudative dermatitis), and also underlying disorders (such as poorly controlled insulin-dependent diabetes mellitus or cancer) are some of the risk factors for secondary infection with S . aureus . In S . aureus-infected primary skin disorders (impetigo, recurrent eczema), 2% mupirocin ointment has proved effective in several clinical trials . S . aureus is responsible for 25% of all burn-wound infections, and burn units could be the point of entry and source of spread of methicillin-resistant S . aureus infection outbreaks . Mupirocin (2% ointment) has also proven effective for topical treatment of these infections . Pressure sores develop in 6% of all patients admitted to acute and chronic health care institutions . An average of three aerobic species (including S . aureus) plus one anaerobic species are isolated when infected . Infectious complications are responsible for 60-80% of all intravenous drug user (IVDU) hospital admissions, 5-20% being due to S . aureus infective endocarditis (IE) . The origin of IE in IVDUs is probably the skin . Data from a Collaborative Spanish Study of IVDU infectious complications (including more than 10,000 episodes) are discussed. J Chemother, 1995 Jul, 7 Suppl 3, 29 - 35 Postoperative wound infections: risk factors and role of Staphylococcus aureus nasal carriage; Perl TM et al.; In the United States the rate of postoperative wound infection varies from one to nine per cent, depending on the surgical procedure . Each postoperative wound infection increases the length of stay in hospital, the cost of the procedure and is associated with significant morbidity . Staphylococcus aureus is the causative agent in 15 to 20% of these infections, although the pathogen isolated varies according to the surgical site . Risk factors for acquiring an infection can be divided into the following categories: host factors, surgical and environmental factors, and microbial characteristics . Host factors which may contribute to an increased risk of infection include: age, prolonged pre-operative length of stay, and concurrent infection at another body site . Increased infection risk may result from an extended surgical procedure, the wound classification, the use of a razor for hair removal before surgery and may also be dependent on the surgeon's technical skill . Microbial factors related to the risk of developing an infection postoperatively are less well defined, however, many outbreaks of surgical wound infections have been linked to personnel carrying an organism which is then transmitted to the patient . Furthermore, patients who carry intranasal S . aureus have a two-to ten-fold increased likelihood of developing a postoperative wound infection due to S . aureus . Identification of patients most at risk of developing an infection is the ultimate goal, however, risk indices must be highly sensitive, specific and accurate . To summarize, the epidemiology of postoperative wound infections remains poorly studied, however, since wound infections contribute significantly to morbidity, mortality and cost, future research is warranted. J Chemother, 1995 Jul, 7 Suppl 3, 19 - 28 Staphylococcus aureus colonisation and bacteraemia in persons infected with human immunodeficiency virus: a dynamic interaction with the host; Craven DE; Bacterial infections are common in persons with symptomatic disease caused by human immunodeficiency virus (HIV) . Colonisation and infection with Staphylococcus aureus is present in 30% to 50% of persons with HIV disease . Risk factors for bacteraemia due to S . aureus include nasal colonisation, advanced HIV disease with CD4+ lymphocyte count < 100/mm3, prior hospitalisations, neutropenia, skin lesions, intravenous drug use, and the presence of invasive devices, such as intravenous catheters . Some antibiotics may increase the risk of S . aureus nasal colonisation, and others such as trimethoprim-sulphamethoxazole and rifabutin may reduce colonisation and disease . Preliminary data suggest that mupirocin may decrease nasal colonisation with S . aureus, but the optimal regimen, duration of effect, use, and concerns about resistance, need further evaluation . Recent data suggest that bacterial infection may accelerate the progression of HIV disease . The presence of bacterial superantigens or cytokines, such as the exotoxins present in many strains of S . aureus have been shown to induce HIV production in human peripheral blood mononuclear leukocyte cultures in vitro, although the precise mechanism is unclear . Thus, HIV infection may increase the risk of S . aureus colonisation and disease and, in turn, infection or colonisation with S . aureus may accelerate the progression of HIV disease to AIDS. J Chemother, 1995 Jul, 7 Suppl 3, 11 - 7 Identifying the patient risk for Staphylococcus aureus bloodstream infections; Espersen F; Staphylococcus aureus is one of the leading causes of bloodstream infection, and these infections still have a high mortality . In certain clinical situations and for the planning of future prophylactic precautions, it is important to identify patients at risk of S . aureus bloodstream infection . Nearly all patients with S . aureus bloodstream infection have underlying conditions such as cardiovascular disease, renal disease, previous surgery, intravenous drug abuse, or malignancy . The great proportion of patients are over 60 years old and the infection is most often acquired in the hospital . The most common focus is intravascular catheters followed by wounds, skin, lungs, bone and joints, and heart valves . The potential risk factors are nasal carriage, previous hospitalisation, surgery, trauma, haemodialysis, presence of high risk focus, acquisition in an orthopaedic ward, and intravenous drug abuse. Microb Pathog, 1995 Jul, 19(1), 49 - 55 Opsonization of Staphylococcus aureus with a fibronectin-binding protein antiserum induces protection in mice; Mamo W et al.; The virulence of Staphylococcus aureus opsonized with an antiserum raised against a recombinant fibronectin-binding protein (FnBP) was compared with homologous, non-opsonized bacteria (treated with pre-immune serum) in a mouse mastitis model . Virulence was evaluated comparing the number of bacteria removed from the infected mammary glands and according to the type of lesions produced . The average number of bacteria recovered from the mammary glands inoculated with S . aureus opsonized with FnBP-antiserum was significantly lower (up to 10(7) cfu/ml) than the average number of bacteria recovered after inoculation with non-opsonized bacteria (up to 10(10) cfu/ml) . Gross examination of infected mammary glands showed that 65% of glands infected with opsonized bacteria developed low grade/or had no pathological changes, and 35% developed severe mastitis whereas, 75% of glands inoculated with non-opsonized bacteria developed severe mastitis and 25% low grade mastitis or had no pathological changes . According to the histopathological examination eight out of 10 glands inoculated with opsonized bacteria produced disseminated focal necrosis or had no pathological changes and two glands produced non reactive necrotic lesions . In contrast, only three out of 10 glands inoculated with non-opsonized homologous bacteria developed disseminated focal necrosis and had no pathological changes while seven glands developed total necrosis. Vet Immunol Immunopathol, 1995 Jul, 47(1-2), 111 - 21 Effects of in vitro supplementation of bovine mammary gland macrophages and peripheral blood lymphocytes with alpha-tocopherol and sodium selenite: implications for udder defences; Ndiweni N et al.; vitamin E and selenium have been observed to enhance the functions of bovine mammary gland macrophages and peripheral blood lymphocytes . In vitro supplementation with these compounds enhanced the production of neutrophil chemotaxins by macrophages stimulated with opsonised Staphylococcus aureus . Supplementation also enhanced the proliferation of peripheral blood lymphocytes in response to stimulation with concanavalin A but not to phytohaemagglutinin or pokeweed mitogen . There was no evidence of additive or synergistic effects of vitamin E and selenium . Results suggest that supplementation of cattle may optimise resistance to mastitis by enhancing the functions of resident macrophage and lymphocyte populations. Ann Vasc Surg, 1995 Jul, 9(4), 378 - 84 Surgical management of infected PTFE hemodialysis grafts: analysis of a 15-year experience; Tabbara MR et al.; The records of 52 consecutive patients who underwent surgical treatment for 57 episodes of hemodialysis graft infection (HGI) from 1977 to 1993 were reviewed to determine the mortality and morbidity associated with this complication and to clarify guidelines for its management . The study group consisted of 35 women and 17 men whose mean age was 57 years at initial graft placement . Thirty-three (58%) HGIs involved straight grafts in the upper arm, 12 (21%) straight forearm grafts, 11 (19%) loop forearm grafts, and 1 (2%) a loop groin fistula . All of these grafts were constructed with polytetrafluoroethylene (PTFE) . All 57 cases of HGI showed at least local evidence and 41 (72%) caused systemic symptoms . Thirty-seven (65%) HGIs were associated with positive blood cultures . The predominant infecting organism was Staphylococcus, which was isolated alone or in combination with other organisms from 40 (70%) graft or would sites . Seventy-eight percent (31/40) of the staphylococcal infections involved Staphylococcus aureus . The median time from graft implantation to diagnosis of HGI was 7 months (mean 16 months, range 0 to 77 months) and from diagnosis to surgical treatment, 4 days (mean 6 days, range 0 to 26 days) . Initial surgical management consisted of complete excision of all prosthetic material in 43 (75%) cases and partial excision in 14 . The 30-day mortality rate following the last operation for the treatment of HGI was 12% (6/52) and was not significantly increased by incomplete excision . Six (86%) of the early deaths were related to sepsis and each of these patients had positive blood cultures.(ABSTRACT TRUNCATED AT 250 WORDS) J Hosp Infect, 1995 Jul, 30(3), 201 - 10 Infection resistance of surface modified catheters with either short-lived or prolonged activity; Sampath LA et al.; It has been suggested that the invasion of microbes into the catheter tract occurs mainly at the time of catheter insertion . To investigate whether the presence of an antimicrobial environment during the initial period after insertion is sufficient to reduce the risk of subsequent catheter colonization and infection, we evaluated the use of benzalkonium chloride-heparin bonded (BZK-hep) central venous catheters, which exhibit short-lived surface antimicrobial activity, using a rat subcutaneous model . Bacterial adherence on these catheters was determined, seven days after challenging the insertion site with 10(6) cfu of Staphylococcus aureus . A chlorhexidine-silver sulphadiazine impregnated catheter (Arrowg+ard), with longer lasting surface antimicrobial activity, and a hydrophilic coated catheter ('Hydrocath'), were evaluated simultaneously for comparison . Unlike Arrowg+ard antiseptic catheters, BZK-hep 'Hydrocath' and control catheters had significant bacterial adherence on their surface . Arrowg+ard catheters were colonized in 19% of the animals compared with 100% in all the other groups (P < 0.05; mean cfu cm-2: control = 1.3 x 10(6), BZK-hep = 4.3 x 10(5), Hydrocath = 2 x 10(5), Arrowg+ard = 71) . Our results indicate that catheters with short-lived surface antimicrobial activity are unlikely to provide long-term protection against catheter-related infection . The efficacy of Arrowg+ard catheters may be due to the initial high rate of kill and prolonged antimicrobial activity. Ann Pharmacother, 1995 Jul-Aug, 29(7-8), 694 - 7 Recurrent methicillin-resistant Staphylococcus aureus osteomyelitis: combination antibiotic therapy with evaluation by serum bactericidal titers; Aspinall SL et al.; OBJECTIVE: To report on a patient with recurrent methicillin-resistant Staphylococcus aureus (MRSA) osteomyelitis and bacteremia successfully treated with combination antibiotic therapy . CASE SUMMARY: Two sets of blood cultures from a 55-year-old man with fever, malaise, and low back pain grew MRSA . Radiologic studies of the spine showed bony changes consistent with osteomyelitis . Soon after completing 6 weeks of vancomycin, the patient experienced a recurrence of back pain . Laboratory values included an increase in the sedimentation rate to 53 mm/h and positive blood cultures for MRSA . Vancomycin, gentamicin, and rifampin were administered for 8 weeks . Serum inhibitory and bactericidal titers were more than 1:1024 for both the peak and trough concentrations . Radiologic studies of the spine showed healing osteomyelitis . Two years after completion of antibiotic therapy, the infection has not recurred . DISCUSSION: Antibiotic therapy alone was attempted because the patient was considered a risky surgical candidate . Serum inhibitory and bactericidal titers documented the high in vivo activity of the vancomycin, gentamicin, and rifampin combination . Initiation of vancomycin, gentamicin, and rifampin combination . Initiation of vancomycin therapy led to disappearance of the fever and back pain . Cure was documented by sustained normalization of the erythrocyte sedimentation rate and radiologic evidence of healing . CONCLUSIONS: Combination antibiotic therapy with vancomycin, rifampin, and low-dose gentamicin (1 mg/kg q12h) may be useful for deep-seated tissue infection caused by MRSA. Infect Control Hosp Epidemiol, 1995 Jul, 16(7), 405 - 11 Control of methicillin-resistant Staphylococcus aureus in a hospital and an intensive care unit; Hartstein AI et al.; OBJECTIVE: To describe methicillin-resistant Staphylococcus aureus (MRSA) control in a hospital, including a surgical intensive care unit (SICU) outbreak . DESIGN: Prospective surveillance of newly identified patients with MRSA . Barrier isolation (disposable gloves for direct contact with patient or immediate environment) was used for the routine care of hospitalized MRSA patients as of October 1991 . Beginning in 1992, MRSA isolates were typed by restriction endonuclease enzyme analysis of plasmid DNA (REAP) and/or pulsed-field gel electrophoresis of genomic DNA (PFGE) . Surveillance information and MRSA typing were used concurrently to identify nosocomial case clustering, confirm cross-infection, and support a need for additional outbreak control interventions . SETTING: University-affiliated public hospital . PARTICIPANTS: Patients with newly identified MRSA colonization or infection from 1991 through 1993 and epidemiologically associated staff providing care to eight SICU patients in an outbreak . INTERVENTIONS: Barrier isolation for affected and unaffected patients in and admitted to the SICU institution when the outbreak was identified and cross-infection confirmed . Anterior nares cultures of staff in contact with outbreak cases for detection of MRSA colonization . RESULTS: Fifty-six hospitalized patients with community-acquired MRSA and 80 patients with nosocomial MRSA colonization or infection were identified during the 3 years . After the introduction of barrier isolation, the annual frequency of new nosocomial MRSA cases decreased and only one outbreak (eight cases in the SICU) caused by type-related isolates occurred . The other 35 nosocomial cases of MRSA during 1992 and 1993 were not epidemiologically related or were caused by isolates with different types . The SICU outbreak ended after instituting barrier isolation for all patients (with and without MRSA) in and admitted to the unit . Six colonized SICU staff were identified . All outbreak cases had identical or related MRSA types by PFGE and REAP . Staff isolates were different from case isolates by typing, and staff were not restricted and not given treatment for colonization . After more than 6 months of follow up, no further outbreaks of MRSA in the SICU or elsewhere in the hospital occurred despite returning to barrier isolation for affected patients only . CONCLUSION: MRSA in hospitals and outbreaks of MRSA in ICUs can be controlled by surveillance and minimal barrier interventions . REAP or PFGE typing of MRSA can be used to support or refute the presence of cross-transmission . Typing also may be helpful when planning and assessing the effectiveness of interventions directed at endemic, as well as outbreak, MRSA control. Infect Control Hosp Epidemiol, 1995 Jul, 16(7), 385 - 90 A statewide surveillance system for antimicrobial-resistant bacteria: New Jersey; Paul SM et al.; OBJECTIVES: To determine the validity of an active, hospital laboratory isolate-based surveillance system in estimating rates of infection and to evaluate the use of surveillance data in describing institutional risk factors for increased rates of infection . Methicillin-resistant Staphylococcus aureus (MRSA) was chosen as the prototype organism for these evaluations . DESIGN: Correlation Study: linear regression analysis and Student's t test were used to evaluate the correlation between number of MRSA isolates and number of MRSA infections in acute-care hospitals . Cross-Sectional Study: Student's t test, analysis of variance, and multiple linear regression analysis were used to evaluate the association between mean annual rate of MRSA blood isolates and institutional risk factors for increased rates of infection . SETTING: Acute-care hospitals, New Jersey . RESULTS: The number of MRSA blood isolates was significantly correlated with MRSA blood infections (R, 0.78; P < .01) and provided a good proxy measure for number of infections . Multivariate analysis demonstrated hospital location in the inner city (P = .02) and number of occupied beds (P < .01) to be independently associated with increased mean annual rates of MRSA blood isolates in acute-care hospitals . CONCLUSIONS: This surveillance system is a valid tool for the estimation of institutional rates of infection and for the determination of institutional risk factors for increased rates of infection . It is ideal for further population-based investigations of antimicrobial-resistant bacteria. Br J Anaesth, 1995 Jul, 75(1), 66 - 70 Inhibition of phagocytosis and killing of bacteria by anaesthetic agents in vitro; Krumholz W et al.; Polymorphonuclear leucocytes (PMNL) are an essential contribution to protection from bacterial infection . We have examined the effects of thiopentone, etomidate, ketamine and flunitrazepam on phagocytosis and killing of Staphylococcus aureus and Escherichia coli by PMNL in vitro with fluorescence microscopy . All anaesthetic agents significantly inhibited both phagocytosis and bactericidal activity . The additives in the commercial preparations may have contributed to the suppression. J Med Assoc Thai, 1995 Jul, 78 Suppl 1, S11 - 4 An outbreak of methicillin-resistant Staphylococcus aureus (M.R.S.A.) in a burn unit; Danchivijitr S et al.; An outbreak of methicillin-resistant Staphyllococcus aureus (MRSA) in the burn unit of Siriraj Hospital from August 1990 to July 1991 was reported . Temporary decrease in the incidence of MRSA was observed during a period when no new cases were admitted . The incidence rose again after normal service resumed . Surveillance cultures in 29 patients from May to July 1991 grew MRSA in 19 patients (65.5 per cent) . Infection by MRSA was found in 14 patients (48.3 per cent) . Methicillin-resistant Staphylococcus aureus first appeared in these patients mainly during the first and second weeks of admission . Wound infections were the only manifestation in 14 patients . Four of the MRSA infected patients died of infection by organisms other than MRSA . Nasal carriage was found in 2 of 25 ward staff members . The cost of treatment for each episode of MRSA infection was as high as 19,322.90 baht . The epidemic of MRSA infections persisted despite all control measures resulting in temporary closure of the ward. J Clin Microbiol, 1995 Jul, 33(7), 1909 - 11 Evidence that the National Committee for Clinical Laboratory Standards disk test is less sensitive than the screen plate for detection of low-expression-class methicillin-resistant Staphylococcus aureus; Mackenzie AM et al.; A low-expression-class methicillin-resistant Staphylococcus aureus strain (strain 9302-2) was sent to 207 laboratories as part of the bacteriology component of the Laboratory Proficiency Testing Program of Ontario . An incorrect (susceptible) result was reported by 16 of 76 (21%) of laboratories that used the National Committee for Clinical Laboratory standards disk test, whereas 1 of 104 laboratories that used other methods reported an incorrect result (P < 0.05) . Experiments showed discrepancies in the disk test results given by Mueller-Hinton agars from three manufacturers . We advise that laboratories should use a low expression-class methicillin-resistant S . aureus isolate as a control for the National Committee for Clinical Laboratory Standards disk test. J Appl Bacteriol, 1995 Jul, 79(1), 69 - 72 The differentiation of Staphylococcus aureus from other Micrococcaceae isolated from bovine mammary glands; Lam TJ et al.; Haemolysin production, the slide coagulase test and the tube coagulase test were assessed for their capability to differentiate Staphylococcus aureus among other Micrococcaceae in 199 isolates from udders of cows in herds with a low bulk milk somatic cell count . The API-Staph test was used as a reference . Haemolysin production was less effective in identifying Staph . aureus among Micrococcaceae than a combination of other tests . Differences were found in the predictive values of results from diagnostic protocols in which the slide coagulase test was performed on all Micrococcaceae, or on beta-haemolysin-negative Micrococcaceae only . Diagnostic protocols in which haemolysin production was combined with the results of the other tests resulted in excellent diagnostic performance and a reduction in diagnostic procedures . Recommendations for routine Staph . aureus identification in bovine mastitis bacteriology are given. Biometals, 1995 Jul, 8(3), 197 - 204 Energetic basis of cadmium toxicity in Staphylococcus aureus; Tynecka Z et al.; In washed cells of cadmium-sensitive Staphylococcus aureus 17810S oxidizing glutamate, initial Cd2+ influx via the Mn2+ porter down membrane potential (delta psi) was fast due to involvement of energy generated by two proton pumps--the respiratory chain and the ATP synthetase complex working in the hydrolytic direction . Such an unusual energy drain for rapid initial Cd2+ influx is suggested to be due to a series of toxic events elicited by Cd2+ accumulation down delta psi generated via the redox proton pump: (i) strong inhibition of glutamate oxidation accompanied by a decrease of electrochemical proton gradient (delta mu H+) formation via the respiratory chain, (ii) automatic reversal of ATP synthetase from biosynthetic to hydrolytic mode, which was monitored by a decrease of delta mu (H+)-dependent ATP synthesis, (iii) acceleration of the initial Cd2+ influx down delta psi generated by the reversed ATP synthetase, the alternative proton pump hydrolyzing endogenous ATP . The primary, cadmium-sensitive targets in strain 17810S seem to be dithiols located in the cytoplasmic glutamate oxidizing system, prior to the membrane-embedded NADH oxidation system . Inhibition by Cd2+ of delta mu (H+)-dependent ATP synthesis and of pH gradient (delta pH)-linked {14C}glutamate transport is a secondary effect due to cadmium-mediated inhibition of delta mu H+ generation at the cytoplasmic level . In washed cells of cadmium-resistant S . aureus 17810R oxidizing glutamate, Cd2+ accumulation was prevented due to activity of the plasmid-coded Cd2+ efflux system . Consequently, delta mu (H+)-producing and -requiring processes were not affected by Cd2+. Rev Esp Cardiol, 1995 Jul, 48(7), 496 - 8 {Aortic prosthetic endocarditis and periprosthetic abscess caused by Staphylococcus aureus}; Garcia Garcia JM et al.; Prosthetic endocarditis with annular abscess formation is a severe complication of cardiac valve replacement fortunately uncommon, though highly lethal . Increasing surgical experience and the high mortality with medical management have led to a widespread recommendation for early prosthetic replacement . We report a case of a 49 year old man with infective endocarditis due to Staphylococcus aureus in aortic ascendens prosthetic and aortic valve prosthetic complicated with periaortic abscess which was as successful treatment by drain of abscess without prosthetic replacement. Gen Pharmacol, 1995 Jul, 26(4), 855 - 64 Potentiation by endothelin-1 of Ca2+ sensitivity of contractile elements depends on Ca2+ influx through L-type Ca2+ channels in the canine cerebral artery; Tanaka Y et al.; 1 . Endothelin-1 (ET-1) contracted canine cerebral artery in a concentration-dependent manner with an increase in intracellular Ca2+ concentration ({Ca2+}i), and at higher concentrations it produced a greater contraction with a smaller increase in {Ca2+}i . 2 . Ca2+ channel antagonist such as d-cis-diltiazem inhibited the tension more effectively than the {Ca2+}i increased by ET-1 . 3 . In Ca(2+)-free solution containing 0.2 mM EGTA, ET-1 elicited a transient increase in {Ca2+}i and tension . 4 . In the Staphylococcus aureus alpha-toxin-permeabilized artery, ET-1 shifted the pCa-tension relationship leftwards in the presence of GTP . 5 . These findings suggest that ET-1 contracts the canine cerebral artery by increasing not only the Ca2+ influx through L-type Ca2+ channels, but also Ca2+ release from the intracellular storage sites, and also Ca2+ sensitivity of contractile elements . The degree of Ca2+ sensitivity is strongly affected by {Ca2+}i which is increased by the Ca2+ influx through L-type Ca2+ channels. Arch Dermatol, 1995 Jul, 131(7), 829 - 32 Superantigens . Do they have a role in skin diseases? Skov L, Baadsgaard O. Superantigens are a group of bacterial and viral proteins that are characterized by their capacity to stimulate a large number of T cells . They bind directly to the major histocompatibility complex class II molecule on the antigen-presenting cell and cross-link the antigen-presenting cell with T cells expressing certain T-cell receptors, leading to polyclonal T-cell activation . They have been shown to play a role in toxic shock syndrome and mucocutaneous lymph node syndrome and are postulated to play a role in other systemic diseases . Because inflammatory skin diseases such as atopic dermatitis and psoriasis are often known to be colonized with superantigen-releasing Staphylococcus aureus, the role of superantigens in skin diseases is of major importance . Recent studies have demonstrated that if a staphylococcal superantigen is applied on intact human skin, a clinical picture of dermatitis evolves . Furthermore, in the presence of superantigens, epidermal cells potently activate T cells . Thus, superantigens may play a role in the induction and exacerbation of inflammatory skin diseases. Am J Kidney Dis, 1995 Jul, 26(1), 47 - 53 Peritonitis in an urban peritoneal dialysis program: an analysis of infecting pathogens; Korbet SM et al.; We have previously found that race, level of education, and peritoneal dialysis system are factors that significantly and independently influence peritonitis rates in our patient population . We now extend these observations by assessing the pathogens responsible for peritonitis in these subgroups . Between January 1, 1981, and May 15, 1993, 248 peritoneal dialysis patients underwent dialysis at our facility . The rate of peritonitis by pathogen was determined in these patients using the fixed effects Poisson model . Total peritonitis rates in black patients (1.89 episodes/patient-year) were significantly greater compared with white patients (1.11 episodes/patient-year; P < 0.0001) . Increased infection rates in black patients were significant for Staphylococcus epidermidis, Staphylococcus aureus, and gram-negative pathogens . The level of education had a negative correlation with peritonitis rates (< or = 8 years, 2.00 episodes/patient-year; 9 to 12 years, 1.64 episodes/patient-year; and > or = 13 years, 1.24 episodes/patient-year) with patients having > or = 13 years of education at the start of dialysis demonstrating a significantly lower total peritonitis rate compared with patients with 9 to 12 years (P = 0.001) or < or = 8 years (P < 0.001) of education . This was accounted for by a significant decrease in infection rates for S epidermidis, polymicrobial, and gram-negative organisms . Finally, patients on automated peritoneal dialysis had significantly lower total peritonitis rates (0.59 episodes/patient-year) compared with patients on either a connect (2.11 episodes/patient-year) or disconnect (1.46 episodes/patient-year) system.(ABSTRACT TRUNCATED AT 250 WORDS) Cell Immunol, 1995 Jul, 163(2), 268 - 79 Expression and biological activities of bovine interleukin 4: effects of recombinant bovine interleukin 4 on T cell proliferation and B cell differentiation and proliferation in vitro; Estes DM et al.; Interleukin 4 (IL-4) is a pleotropic cytokine affecting a wide range of cell types in both the mouse and the human . These activities include regulation of the growth and differentiation of both T and B lymphocytes . The activities of IL-4 in nonprimate, nonmurine systems are not well established . Herein, we demonstrate in the bovine system that IL-4 upregulates production of IgM, IgG1, and IgE in the presence of a variety of costimulators including anti-IgM, Staphylococcus aureus cowan strain I, and pokeweed mitogen . IgE responses are potentiated by the addition of IL-2 to IL-4 . Culture of bovine B lymphocytes with IL-4 in the absence of additional costimulators resulted in the increased surface expression of CD23 (low-affinity Fc epsilon RII), IgM, IL-2R, and MHC class II in a dose-dependent manner . IL-4 alone increased basal levels of proliferation of bulk peripheral blood mononuclear cells but in the presence of Con A inhibited proliferation . In contrast to the activities of IL-4 in the murine system, proliferation of TH1- and TH2-like clones was inhibited in a dose-dependent manner as assessed by antigen-or IL-2-driven in vitro proliferative responses . These observations are consistent with the role of IL-4 as a key player in regulation of both T and B cell responses. J Lab Clin Med, 1995 Jul, 126(1), 29 - 35 Molecular typing of the methicillin resistance determinant (mec) of clinical strains of Staphylococcus based on mec hypervariable region length polymorphisms; Nishi J et al.; We used a method for molecular typing the methicillin resistance determinant (mec) based on the size of the mec-associated hypervariable region amplified by the polymerase chain reaction (PCR) to examine 61 methicillin-resistant Staphylococcus aureus (MRSA), 15 methicillin-resistant (Mcr) S . epidermidis, and 11 Mcr S . haemolyticus clinical isolates . In the 61 MRSA isolates, five sizes of PCR products were observed . The MRSA isolates were grouped into five hypervariable region (HVR) genotypes on the basis of the size of the PCR product . Three different sizes were detected among 15 isolates of Mcr S . epidermidis and two sizes among 11 isolates of Mcr S . haemolyticus . The PCR products amplified from 14 of 15 Mcr S . epidermidis isolates were the same as products amplified from MRSA isolates, which was confirmed by the PCR-SSCP (single-strand conformation polymorphism) method . In methicillin-susceptible isolates, the target gene was not amplified . This method is thought to be useful in epidemiologic investigations of nosocomial infections caused by MRSA . This is the first typing method capable of comparing the mec determinants of MRSA isolates and Mcr coagulase-negative staphylococcal isolates to establish the origin of the mec determinant. Crit Care Med, 1995 Jul, 23(7), 1200 - 3 Significant reduction in methicillin-resistant Staphylococcus aureus ventilator-associated pneumonia associated with the institution of a prevention protocol; Rumbak MJ et al.; OBJECTIVE: To determine whether the institution of a methicillin-resistant Staphylococcus aureus prevention protocol was associated with a decrease in methicillin-resistant S . aureus ventilator-associated pneumonia in long-term, acute care ventilator patients . DESIGN: A retrospective chart review comparing the number of episodes of clinical pneumonia per patient ventilator day in the 12 months preceding and 24 months following the introduction of the protocol . SETTING: University affiliated, long-term, acute care ventilator hospital . PATIENTS: Long-term, acute care ventilated patients who presented with clinical pneumonia . INTERVENTIONS: Addition of a methicillin-resistant S . aureus prevention protocol . In addition to universal precautions, the protocol consisted of mupirocin 2% ointment applied to the anterior nares, and whole body washing with chlorhexidine . All patients were given mupirocin and chlorhexidine twice weekly . Patients were cohorted in the same room if they were, or had been, infected or colonized with methicillin-resistant S . aureus in any anatomical location or at any time . This procedure replaced strict isolation of methicillin-resistant S . aureus-infected or colonized individuals . MEASUREMENTS AND MAIN RESULTS: Clinical pneumonia was diagnosed when a patient developed fever, bronchorrhea, increased white blood cell count, methicillin-resistant S . aureus isolated from the tracheal aspirate, and new or increasing infiltrate on chest roentgenograph . During the 12 months preceding the protocol, there were 0.2% episodes of methicillin-resistant S . aureus ventilator-associated pneumonia per ventilated patient day compared with 0.026% in the 24 months after the protocol (p < .001) . The relative and absolute risk reductions associated with the introduction of the protocol were 87% and 6, respectively . CONCLUSIONS: The period following the institution of the protocol showed a significant reduction in episodes of clinical pneumonia compared with the 12-month period preceding the use of the protocol (p < .001) . Thus, we conclude that the introduction of this protocol is associated with a significant decrease in methicillin-resistant S . aureus ventilator-associated pneumonia. J Anim Sci, 1995 Jul, 73(7), 2079 - 85 Effect of copper depletion and repletion on lymphocyte blastogenesis and neutrophil bactericidal function in beef heifers; Arthington JD et al.; Thirty-two beef heifers were used to examine the effect of dietary copper depletion and repletion on neutrophil and lymphocyte functions . Heifers allotted to the control group (C+; n = 8) were fed a basal roughage/concentrate diet with Cu-sulfate supplementation (Cu > or = 8 ppm) . To induce a Cu deficiency (depletion phase d 0 to 60), treated (T; n = 24) heifers received a diet supplemented with sulfur (.3% of diet) and sodium molybdate to achieve a Cu:Mo ratio of 1:1.5 . Liver biopsies were collected on d 0, 27, and 60 . Despite random allocation, T heifers had lower initial liver Cu concentrations (P < .01) than C+ heifers . At the start of the repletion phase (d 0, equal to d 60 of depletion), treated heifers were allotted by liver Cu concentration to three treatments (n = 8/treatment): Cu sulfate (S; Cu = 10 ppm), Cu proteinate (P; Cu = 10 ppm), or a negative control (C-) that remained on Mo and S supplementation . During the repletion phase, livers were biopsied on d 0, 14, and 45 . By d 45, both S and P heifers had greater (P < .05) liver Cu concentrations than C- heifers . For both depletion and repletion phases, no treatment differences were detected in liver Mo or S concentrations . Jugular blood was collected on d 0, 27, and 55 of the depletion phase and d 0, 13, and 42 of the repletion phase . Neutrophils were isolated and incubated with Staphylococcus aureus to determine neutrophil bactericidal capacity (NBC).(ABSTRACT TRUNCATED AT 250 WORDS) Clin Diagn Lab Immunol, 1995 Jul, 2(4), 412 - 6 Intrinsic defect in B cells of patients with hyper-immunoglobulin M syndrome; Porat YB et al.; We challenge the theory that the CD40-CD40 ligand is the only explanation for X-linked immunodeficiency in patients with hyper-immunoglobulin M (IgM) syndrome (HIGM1), and we demonstrate an intrinsic defect in the patients' B cells . Patients with HIGM1 have a defective CD40 ligand on their activated T-helper cells; therefore, they cannot receive signals for isotype switching when the cells are activated by T cell-dependent antigens . We activated mononuclear cells from three patients with HIGM1 and from three healthy blood donors with T cell-independent mitogens and studied their proliferative responses and Ig secretion . Normal murine plasma membrane fragments were implanted into peripheral blood mononuclear cells, and the cells were activated with Staphylococcus aureus Cowan I, pokeweed mitogen, and lipopolysaccharide . This implantation significantly augmented the proliferative responses to the mitogens in two patients . However, it augmented IGM secretion in response to B-cell mitogens in only one patient . No IgG or IgA response could be detected in the implanted mononuclear cells that originated from patients with HIGM1, unlike implanted mononuclear cells from healthy donors, which responded by IgM, IgG, and IgA antibody secretion following their stimulation with B-cell mitogens . The data suggest that the B cells of patients with HIGM1 possess an additional defect which prevents Ig isotype switching in response to T cell-independent mitogens . This defect is not located in the membrane receptors or within the membrane enzymes. Cell Mol Biol (Noisy-le-grand), 1995 Jul, 41(5), 615 - 23 Nucleic acid amplification and related techniques in microbiological diagnostics and epidemiology; Van Belkum A et al.; The use of nucleic acid amplification techniques within the medical microbiology laboratory is becoming more and more accepted . Polymerase chain reaction (PCR) tests or nucleic acid sequence based amplification (NASBA) assays are already available in the form of commercial kits . Although the technology has been adapted for application in a routine diagnostic setting, some of the systems' characteristics are still amenable to improvement . In this communication several of these aspects will be discussed . Reproducibility of DNA amplification mediated diagnostics and quality control of tests aiming at detection or genetic typing of both viral and bacterial microorganisms, will be discussed . This will be exemplified by the results obtained in multicenter studies on PCR diagnostics of the hepatitis viruses HBV and HCV and by data gathered in the course of PCR mediated DNA fingerprinting of Staphylococcus aureus strains, also performed in different institutes . Application of related techniques such as direct sequencing of amplified (c)DNA or the development of species-specific DNA probes will be described. Acta Anaesthesiol Scand, 1995 Jul, 39(5), 624 - 7 The effects of midazolam, droperidol, fentanyl, and alfentanil on phagocytosis and killing of bacteria by polymorphonuclear leukocytes in vitro; Krumholz W et al.; Polymorphonuclear leukocytes (PMNL) make an outstanding contribution to the defence against invading bacteria . We studied the effects of midazolam, droperidol, fentanyl, and alfentanil on phagocytosis and killing of Staphylococcus aureus and Escherichia coli by PMNL in vitro . Using a method described by Eggleton et al., PMNL were isolated from venous blood samples obtained from ten volunteers . The fluorescence microscopic method mentioned by Bellinati-Pires et al . was used to examine phagocytosis and killing . Whereas both midazolam and droperidol caused a significant inhibition of phagocytosis as well as bactericidal activity, fentanyl and alfentanil did not influence these PMNL functions . In order to find out whether midazolam and droperidol are able to intensify perioperative bacterial infections in vivo as well, additional clinical studies should be conducted. Vet Microbiol, 1995 Jul, 45(2-3), 275 - 9 The amino acid requirements of Staphylococcus aureus isolated from cases of bovine mastitis; Lincoln RA et al.; The amino acid requirements of seven strains of Staphylococcus aureus isolated from cases of bovine mastitis were determined . Arginine, cystine, glycine, leucine, proline and valine were essential for the growth of all isolates . In addition, all isolates required one or more of the following: glutamic acid, histidine, isoleucine, lysine, methionine, phenylalanine, tryptophan and tyrosine. Vet Microbiol, 1995 Jul, 45(2-3), 139 - 50 Evaluation of phenotypic and genotypic methods for epidemiological typing of Staphylococcus aureus isolates from bovine mastitis in Denmark; Aarestrup FM et al.; The value of five different typing methods (antibiogram typing, biotyping, phage typing, plasmid profiling and restriction fragment length polymorphism of the gene encoding 16S and 23S ribosomal RNA (ribotyping)), in discriminating 105 Staphylococcus aureus strains from bovine milk samples obtained from 105 different Danish dairy herds was investigated . A total of 85 strains (81%) proved susceptible to all of the 11 antibiotics tested, and the remaining 20 strains could be divided into 5 different antibiogram patterns . The predominant resistance pattern, penicillin resistance, was observed in 15 (75%) of the 20 antibiotic resistant strains . Biotyping assigned the strains to 14 different types, with the most common type accounting for 25.7% of the strains . Ninety eight (93.3%) strains could be typed by phages, assigning them to 19 different phage types . The predominant phage type accounted for 31.4% of the strains . Eight different plasmid profiles was observed among 24 (23%) strains harbouring plasmids . Ribotyping yielded 30 different types, with the most common accounting for 29.5% of the strains . The single most discriminatory typing method was ribotyping (0.863) followed by biotyping (0.842) and phage typing (0.795) . Plasmid profiling (0.395) and antibiogram typing (0.327) had low discriminatory indices . Correspondence among ribotypes and the presence or absence of plasmids were observed, as was some degree of correspondence between ribotype, phage type and biotype . In general the correspondence between phage type and ribotype were stronger than between biotype and ribotype and between biotype and phage type . All combinations of two or more methods led to an improved index of discrimination compared to the individual methods indicating, that some subdivision of types had taken place . The combination of phage, bio- or ribotyping or all three methods in combination are considered to be an efficient combination of typing methods for epidemiological investigation of S . aureus mastitis. J Obstet Gynecol Neonatal Nurs, 1995 Jul-Aug, 24(6), 557 - 61 Toxic shock syndrome: an opportunity for nursing intervention; Creehan PA; Toxic Shock Syndrome (TSS) is a potentially fatal illness caused by a particular strain of Staphylococcus aureus . The clinical presentation is similar to that of septic shock . The incidence of TSS peaked in the late 1970s and early 1980s, probably as a result of availability of super absorbent tampons . Although most commonly associated with menstruation, the overall incidence of menstrual and nonmenstrual TSS in men and women ranges from 1 to 3 per 100,000 . There are almost equal numbers of menstrual and nonmenstrual cases of TSS identified annually . S aureus, the causative microorganism in cases of TSS, has been isolated from many body tissues . Toxic shock syndrome presents as a flu-like illness with high fever, vomiting, diarrhea, general malaise, and muscle weakness . Nursing and medical management focus on controlling or preventing potentially serious complications, such as adult respiratory distress syndrome, renal failure, electrolyte imbalances, disseminated intravascular coagulation, encephalopathy, and cardiomyopathy . Judicious use of tampons and barrier contraceptive devices may decrease the risk of developing TSS. J Med Assoc Thai, 1995 Jul, 78 Suppl 2, S78 - 80 Surgical wound infections in gynaecology at Rajvithi Hospital 1989-1990; Choojitr W et al.; A survey on surgical wound infections in gynaecological patients at Rajvithi Hospital was done . One thousand three hundred and eighty-three cases who were operated on from 1989 to 1990 were included . The overall rate of surgical wound infection was 4.92% . Vaginal stump infection accounted for 52.9%, and abdominal wound infections 30.9% . Aerobic gram negative bacilli were the main group of isolated bacteria (45.12%) followed by aerobic gram positive cocci (19.51%) . Methicillin resistant Staphylococcus aureus (MRSA) were found in 3.66% of the isolates . Antimicrobials were used in combination . Two antimicrobials were precribed in 4.84%, 3 drugs in 17.74%, 4 drugs in 25.81% and more than 4 drugs in drugs in as many as 51.61% of the patients. Nippon Kyobu Geka Gakkai Zasshi, 1995 Jul, 43(7), 1073 - 6 {Intrathoracic transposition of the musculocutaneous flap in treating empyema}; Nomori H et al.; A 62-year-old woman suffered right empyema caused by methicillin-resistant staphylococcus aureus (MRSA) which occurred following a pancreatico-duodenectomy . After open drainage thoracotomy, intrathoracic transposition of the extended musculocutaneous (MC) flap of the latissimus dorsi was performed . The patient is now in good health, without recurrence of either the empyema or the carcinoma, 19 months after the operation . The MC flap, compared to muscle flap, has the advantages that (1) larger empyema cavities can be obliterated, and (2) the deformity of the thoracic wall can be minimized, because of the small range of resected rib segments and well-preserved volume of subcutaneous tissue in the flap long after transposition. Kansenshogaku Zasshi, 1995 Jul, 69(7), 835 - 9 {A case of myelodysplastic syndrome who died of septic pulmonary embolism}; Nakamura A et al.; A case of myelodysplastic syndrome (MDS) complicated by septic pulmonary embolism is reported . A 61-year-old female who had been followed for refractory anemia with excess of blasts suddenly died of acute respiratory failure . An autopsy revealed massive pulmonary emboli with gram-positive cocci gathered in the emboli and alveolar spaces . Staphylococcus aureus was also detected through a blood culture from the right atrium . We speculate that pulmonary embolism was the result of septicemia induced by the immunosuppressive condition associated with MDS. Int J Pept Protein Res, 1995 Jul, 46(1), 69 - 72 Primary structure of a cytotoxin-like basic protein from Naja naja naja (Indian cobra) venom; Babu AS et al.; The complete amino acid sequence of a cytotoxin-like basic protein (CLBP) from the venom of Naja naja naja (Indian Cobra) was determined by manual degradation using a 4-dimethylaminoazobenzene-4'-isothiocyanate double-coupling method . Peptide fragments obtained by chemical cleavage with cyanogen bromide and enzymic cleavages with trypsin and Staphylococcus aureus proteases for sequence analysis were purified by reversed-phase chromatography . The total number of amino acid residues was 61, with leucine as the C-terminal residue. J Am Soc Echocardiogr, 1995 Jul-Aug, 8(4), 554 - 6 Primary left ventricular mural endocarditis diagnosed by transesophageal echocardiography; Shirani J et al.; Primary left ventricular mural abscess was detected by transesophageal echocardiography and was confirmed at necropsy in a 44-year-old woman with Staphylococcus aureus bacteremia and cerebrovascular embolism . In two occasions, transthoracic echocardiography failed to show the mural abscess in this patient . Because of the aggressive nature of primary mural endocarditis, early use of transesophageal echocardiography is recommended in patients with Staphylococcal bacteremia and suspected endocarditis even in the absence of valvular abnormalities detectable by the transthoracic approach. J Clin Microbiol, 1995 Jul, 33(7), 1769 - 74 Phenotypic characterization of epidemic versus sporadic strains of methicillin-resistant Staphylococcus aureus; Van Wamel WJ et al.; Forty strains of methicillin-resistant Staphylococcus aureus (MRSA) were divided on the basis of their epidemiologic behavior into two subgroups, sporadic MRSA (SMRSA) and epidemic MRSA (EMRSA) strains . The strains were examined for binding of 125I-labelled fibronectin, vitronectin, collagen, Fc fragments of immunoglobulin G, and fibrinogen . A significant difference between EMRSA and SMRSA strains was found for binding of 125I-labelled fibrinogen and for Fc fragments of immunoglobulin G, (P < 0.05) . No significant difference in the binding of 125I-labelled fibronectin and collagen was found between EMRSA and SMRSA strains . The binding of 125I-labelled vitronectin to MRSA strains was found to be aspecific . Capsular serotypes of the strains were determined with monoclonal antibodies against capsular types 5 and 8 . Strains could be divided into the following four groups: types 5, 8, and 5/8 and nontypeable . More nontypeable strains were found in the EMRSA group (66.6%) . Significantly more EMRSA strains (79%) than SMRSA strains (44%) produced alpha-toxin (P < 0.025) . Logistic regression analysis using a combination of the parameters 125I-labelled immunoglobulin G binding, capsular type, and alpha-toxin production predicted the epidemic character with a sensitivity of 83% and a specificity of 75%. Lett Appl Microbiol, 1995 Jul, 21(1), 1 - 4 Application of a fluorescent redox dye for enumeration of metabolically active bacteria on albumin-coated titanium surfaces; McDowell SG et al.; A bacterial staining method using fluorescent redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) is described for quantifying actively respiring bacteria that adhere to commercially pure titanium surfaces coated with cross-linked albumin . This has not been possible to date using ordinary DNA stains such as propidium iodide (PI) or Hoechst, both of which produce a very bright background . With this technique, it was demonstrated that the cross-linked albumin inhibited the adherence of Staphylococcus aureus and Staph . epidermidis to the titanium surface. J Bacteriol, 1995 Jul, 177(13), 3631 - 40 Localization of penicillin-binding proteins to the splitting system of Staphylococcus aureus septa by using a mercury-penicillin V derivative; Paul TR et al.; Precise localization of penicillin-binding protein (PBP)-antibiotic complexes in a methicillin-sensitive Staphylococcus aureus strain (BB255), its isogenic heterogeneous methicillin-resistant transductant (BB270), and a homogeneous methicillin-resistant strain (Col) was investigated by high-resolution electron microscopy . A mercury-penicillin V (Hg-pen V) derivative was used as a heavy metal-labeled, electron-dense probe for accurately localizing PBPs in situ in single bacterial cells during growth . The most striking feature of thin sections was the presence of an abnormally large (17 to 24 nm in width) splitting system within the thick cross walls or septa of Hg-pen V-treated bacteria of all strains . Untreated control cells possessed a thin, condensed splitting system, 7 to 9 nm in width . A thick splitting system was also distinguishable in unstained thin sections, thereby confirming that the electron contrast of this structure was not attributed to binding of bulky heavy metal stains usually used for electron microscopy . Biochemical analyses demonstrated that Hg-pen V bound to isolated plasma membranes as well as sodium dodecyl sulfate-treated cell walls and that two or more PBPs in each strain bound to this antibiotic . In contrast, the splitting system in penicillin V-treated bacteria was rarely visible after 30 min in the presence of antibiotic . These findings suggest that while most PBPs were associated with the plasma membrane, a proportion of PBPs were located within the fabric of the cell wall, in particular, in the splitting system . Inhibition of one or more high-M(r) PBPs by beta-lactam antibiotics modified the splitting system and cross-wall structure, therefore supporting a role for these PBPs in the synthesis and architectural design of these structures in S . aureus. Eur J Nucl Med, 1995 Jul, 22(7), 638 - 44 Contribution of phagocytic cells and bacteria to the accumulation of technetium-99m labelled polyclonal human immunoglobulin at sites of inflammation; Calame W et al.; The purpose of this study was to assess the contribution of phagocytic cells and bacteria to the accumulation of technetium-99m labelled polyclonal human immunoglobulin (HIG) at sites of inflammation . Mice were intraperitoneally injected with Staphylococcus aureus (SA animals), with heat-inactivated newborn calf serum (NBCS, to mimic a non-bacterial inflammation) or with physiological saline (controls); 1 h thereafter they received HIG . At various intervals after the administration of HIG the mice were killed, and the percentages of radioactivity in the peritoneal effluent and attached to the cellular and bacterial fraction thereof were established . Furthermore, the total number of cells and that of bacteria in the fluid were quantitated . The percentage of activity in the effluent in the SA animals was (P < 0.02) higher than those in the NBCS-injected animals and controls from 4 h onwards . In all groups of mice this percentage was highest at 4 h and decreased (P < 0.01) afterwards . The percentage of cell-bound activity and the total number of cells remained fairly constant or increased with time in the SA animals (P < 0.01) . The bacteria-bound activity remained rather constant throughout the experiment and ranged between 4% and 6% . In the SA-infected animals the percentage of cell-bound activity was correlated with the total number of cells (macrophages but especially neutrophils) but even more strongly with the number of cell-associated bacteria . In the NBCS-injected animals a correlation was demonstrated between the cell-bound activity and the total number of cells (only neutrophils).(ABSTRACT TRUNCATED AT 250 WORDS) Biofizika, 1995 Jul-Aug, 40(4), 786 - 92 {Daily observations (1970-1992) of fluctuations in frequency of occurrence of a sector structure in bacterial colonies selected from open air and from S . aureus cultures}; Faraone P; The frequencies of sector structure occurrence in different bacteria colonies (SSC) denuded from open air every day in period from 1970 to 1982 years and also in laboratory cultures Staphylococcus Aureus from 1984 to 1992 were investigated . The value SSC was expressed in percents to general number of colonies . Variations of average SSC is found out distinctly expressed opposition to 11-years cycle of Solar activity for the same period of time . The year cycle SSC was registered also with local maxima in June, August and November and global minimum in March . SSC also were observed on level of sea, on height about 1000 m over the level of sea and in gallery under the rock of thickness 1400 m (Gran Cacco, National laboratory, Assergi) . The value of SSC has its maximum on height 1000 M and its minimum in gallery under the rock; it is possible, SSC depends on the intensity of space galactic rays . The results of experience, executed in April-July 1992 on cultures S.aureus have compared with average (on ten-day time periods) values for the same months of 1984-1991 . Good correlation of these two curves of SSC was received . Array of measurements received from 1970 to 1992 and made conclusions can be used for long-term forecasts of cosmophysical fluctuating phenomena at least on latitude between Rome and Milano. Antimicrob Agents Chemother, 1995 Jul, 39(7), 1554 - 8 Analysis of gyrA and grlA mutations in stepwise-selected ciprofloxacin-resistant mutants of Staphylococcus aureus; Ferrero L et al.; Fluoroquinolone-resistant mutants were obtained in vitro from Staphylococcus aureus RN4220 by stepwise selection on increasing concentrations of ciprofloxacin . Results from sequence analysis of the quinolone resistance-determining region of GyrA and of the corresponding region of GrlA, the DNA topoisomerase IV subunit, showed an alteration of Ser-80 to Tyr (corresponding to Ser-83 of Escherichia coli GyrA) or Glu-84 to Lys in GrlA of both low- and high-level quinolone-resistant mutants . Second-step mutants were found to have, in addition to a mutation in grlA, reduced accumulation of norfloxacin or an alteration in GyrA at Ser-84 to Leu or Glu-88 to Lys . Third-step mutants derived from second-step mutants with reduced accumulation were found to have a mutation in gyrA . The results from this study demonstrated that mutations in gyrA or mutations leading to reduced drug accumulation occur after alteration of GrlA, supporting the previous findings (L . Ferrero, B . Cameron, B . Manse, D . Lagneaux, J . Crouzet, A . Famechon, and F . Blanche, Mol . Microbiol . 13:641-653, 1994) that DNA topoisomerase IV is a primary target of fluoroquinolones in S . aureus. Antimicrob Agents Chemother, 1995 Jul, 39(7), 1505 - 11 Pharmacodynamics of RP 59500 alone and in combination with vancomycin against Staphylococcus aureus in an in vitro-infected fibrin clot model; Kang SL et al.; The bactericidal activity and emergence of resistance to RP 59500 (quinupristin/dalfopristin) when it was administered alone and in combination with vancomycin against fibrin clots that have been infected with methicillin-susceptible Staphylococcus aureus ATCC 25923 or methicillin-resistant S . aureus (MRSA) 67 were evaluated in an in vitro pharmacodynamic infected fibrin clot model . Fibrin clots were infected with S . aureus to achieve an inoculum of approximately 10(9) CFU/g . Antibiotics were administered to simulate pharmacokinetics in humans: RP 59500 (7.5 mg/kg of body weight) every 8 h and vancomycin (15 mg/kg) every 12 h over 72 h . Preliminary test tube time-kill experiments with an inoculum of approximately 10(5) CFU/ml suggested that RP 59500 was more rapid in achieving a 99.9% reduction in the number of CFU per milliliter than vancomycin against ATCC 25923 (6.94 versus 24 h; P = 0.0003) and MRSA 67 (6.77 versus 17.03 h; P = 0.004) . At a higher inoculum (approximately 10(8) CFU/ml), 99.9% kill was achieved only with the combination regimen against ATCC 25923 and MRSA 67 (10.9 and 10.5 h, respectively), with total reductions of 6.35 and 6.33 log10 CFU/ml over 24 h, respectively . In the fibrin clot model, RP 59500 was more effective than vancomycin in reducing organism titers over 72 h . In the fibrin clot model, the most optimal therapy was the combination regimen. Antimicrob Agents Chemother, 1995 Jul, 39(7), 1419 - 24 Treatment of experimental endocarditis due to erythromycin-susceptible or -resistant methicillin-resistant Staphylococcus aureus with RP 59500; Entenza JM et al.; RP 59500 is a new injectable streptogramin composed of two synergistic components (quinupristin and dalfopristin) which are active against erythromycin-susceptible and -resistant gram-positive pathogens . The present experiments compared the therapeutic efficacy of RP 59500 with that of vancomycin against experimental endocarditis due to either of two erythromycin-susceptible or two constitutively erythromycin-resistant isolates of methicillin-resistant Staphylococcus aureus . RP 59500 had low MICs for the four test organisms as well as for 24 additional isolates (the MIC at which 90% of the isolates were inhibited was < 1 mg/liter) which were mostly inducibly (47%) or constitutively (39%) erythromycin resistant . Aortic endocarditis in rats was produced with catheter-induced vegetations . Three-day therapy was initiated 12 h after infection, and the drugs were delivered via a computerized pump, which permitted the mimicking of the drug kinetics produced in human serum by twice-daily intravenous injections of 7 mg of RP 59500 per kg of body weight or 1 g of vancomycin . Both antibiotics reduced vegetation bacterial titers to below detection levels in ca . 70% of animals infected with the erythromycin-susceptible isolates (P < 0.05 compared with titers in controls) . Vancomycin was also effective against the constitutively resistant strains, but RP 59500 failed against these isolates . Further experiments proved that RP 59500 failures were related to the very short life span of dalfopristin in serum (< or = 2 h, compared with > or = 6 h for quinupristin), since successful treatment was restored by artificially prolonging the dalfopristin levels for 6 h . Thus, RP 59500 is a promising alternative to vancomycin against methicillin-resistant S . aureus infections, provided that pharmacokinetic parameters are adjusted to afford prolonged levels of both of its constituents in serum . This observation is also relevant to humans, in whom the life span of dalfopristin in serum is also shorter than that of quinupristin. Antimicrob Agents Chemother, 1995 Jul, 39(7), 1414 - 8 Incidence of various gyrA mutants in 451 Staphylococcus aureus strains isolated in Japan and their susceptibilities to 10 fluoroquinolones; Takenouchi T et al.; Point mutations in the gyrA genes of 451 clinical strains of Staphylococcus aureus isolated in Japan were detected by a combination of nonradioisotopic single-strand conformation polymorphism analysis and restriction fragment length polymorphism analysis and by direct sequencing . Six types of gyrA mutations were observed in 149 of the 451 strains (33%), and ofloxacin MICs were greater than 6.25 micrograms/ml for 147 of the 149 strains (98.7%) . These mutations were localized between codons 84 and 88, and they were associated with fluoroquinolone resistance . Two types of silent mutations were also found . Among these eight types of mutations, three types are novel, i.e., the serine at position 84 (Ser-84)-->Val (TCA-->GTA), Ser-84-->Leu (TCA-->TTA) plus Ile-86 (ATT-->ATC, silent), and Phe-110 (TTT-->TTC, silent) . Among GyrA mutants, strains with a Ser-84-->Leu alteration and strains with a Glu-88-->Lys alteration were dominant . In contrast, few strains had Ser-84-->Ala and Glu-88-->Gly alterations . All fluoroquinolones tested showed greater than a fourfold decrease in their activities in terms of their MICs that inhibited 50% of strains tested for each GyrA mutant, in comparison with their MICs that inhibited 50% of strains tested for susceptible strains . Most of the currently available fluoroquinolones, such as norfloxacin, enoxacin, ofloxacin, ciprofloxacin, tosufloxacin, lomefloxacin, sparfloxacin, and fleroxacin, were ineffective against each mutant . Mutants containing a Ser-84-->Leu or Val alteration showed high-level resistance to fluoroquinolones, and one containing a Ser-84-->Ala alteration showed relatively low-level resistance . Double mutations were associated with a higher level of resistance than single mutations. Nippon Saikingaku Zasshi, 1995 Jul, 50(3), 871 - 9 {Autobacteriographic studies on the distribution and localization of Staphylococcus aureus in mice}; Tani K; Autobacteriography was proposed as a bacteriological method to follow distribution and localization of bacteria in experimentally infected animals . Infectious organisms were restricted to rifampicin-resistant strains to prevent contamination during autobacteriography . Mice were infected with Staphylococcus aureus Smith diffuse type RFPr (rifampicin-resistant) by the intravenous route and frozen at various intervals after infection . Whole body sections (40-microns thick) of the mice were transferred onto selective agar medium containing rifampicin to incubate at 37 degrees C . On day 1 after infection, dense colonies of the infecting organism on the sections were distributed in the whole body . On day 3, few organisms were detected in the liver and many were observed in the spleen, kidney and intestinal tract . On days 7, 14 and 21, the organisms in the liver and spleen disappeared, and those in the kidney and intestine remained . The remaining infectious organisms were demonstrated in the kidney and intestinal tract by autobacteriography of mice infected with S . aureus Smith compact type RFPr . By cultivation of the homogenate of the gastrointestinal tissues and their contents, the infectious organisms were detected mainly in the lower small intestines, cecum and large intestines. Presse Med, 1995 Jun 24, 24(23), 1075 - 7 {Repeated studies of the prevalence of Staphylococcus aureus in the nasal cavity in hemodialysed patients}; Montagnac R et al.; OBJECTIVES: In order to better establish a prevention strategy based on mupirocin, we evaluated nasal carriage of Staphylococcus aureus in haemodialysis patients over a 15 month period . METHODS: Search for Staphylococcus aureus in the nasal cavities was made every 2 months in 92 chronic dialysis patients . These patients were divided into 3 groups according to the nature of the carriage: non-permanent, intermittent or permanent . RESULTS: Among the 80 patients retained for analysis, there were 27.5% with intermittent carriage and 11.25% with permanent carriage . Factors which appeared to protect against carriage were rural residence and home self-dialysis . CONCLUSION: Repeated long-term search for nasal carriage of Staphylococcus aureus has provided reliable data for each patient and gave information on the effects of epidemiological conditions and health care structures. J Chromatogr A, 1995 Jun 23, 705(1), 135 - 54 Capillary electrophoresis of S . nuclease mutants; Kalman F et al.; The electrophoretic migration behavior of 12 S . nuclease variants from Staphylococcus aureus with small but well defined structural differences from site directed mutation was investigated in free solution capillary electrophoresis at pH 2.8 to 9.5 . The nucleases are basic proteins; the pI and the M(r) of the wild type are 10.3 and 16.811 kd, respectively . With specially selected oligoamino buffers and with an inert, hydrophilic wall coating in 75 microns I.D . quartz capillary tubes, most of the proteins could be separated by CZE without interference by wall adsorption even at pH 9.5 where the selectivity was the highest . At pH 2.8, 4.1 and 7.0, S . nucleases are known to be in the random coil, "swollen" and the tight native state . Assuming that in a given state, i.e., at a certain pH, the molecular radii of the nucleases are the same, their hydrodynamic radii were calculated from their pertinent electrophoretic mobilities . The respective radii of 50.1, 26.8, and 25.0 Angstrum thus obtained agreed very well with the corresponding radii of gyration obtained from X-ray scattering . In fact, from the electrophoretic mobilities at pH 9.5, the existence of a hitherto unknown swollen basic state of the nuclease having a hydrodynamic radius of 30.5 Angstrum was postulated . In addition, a method was described to evaluate the valence of the protein at different pH from their pertinent electrophoretic mobilities . A general advantage of this method is that only the differences between the valences of the mutants and the wild type are needed; and for none of the proteins is required the knowledge of the actual valence . The results of the methods allowed the construction of a pH profile of the protein's valence . For the wild type, this profile was compared to the H+ titration curve and the agreement was excellent . Both methods employed some novel structure-electrophoretic mobility relationships and the predicted protein properties compared remarkably well to the values obtained by exoelectrophoretic methods such as pH titration and X-ray scattering . Surprisingly, certain S . nucleases having the same valence could also be readily separated by CZE in some cases under the same conditions used for the others . Close examination of appropriate X-ray crystallography and/or NMR data indicated subtle differences in the molecular structure of these proteins that could be responsible for slight alteration in their hydrodynamic radii.(ABSTRACT TRUNCATED AT 400 WORDS) Cytometry, 1995 Jun 15, 22(2), 154 - 7 Amphotericin B susceptibility of Candida species assessed by rapid flow cytometric membrane potential assay; Ordonez JV et al.; A method for detection of the susceptibility of Candida species to amphotericin B is described . The technique is a modification of a similar flow cytometric technique designed to detect antibiotic susceptibility of Staphylococcus aureus . Membrane potential changes caused by the antibiotic are measured flow cytometrically by using the fluorescent membrane potential-sensitive dye 3,3'-dipentyloxacarbocyanine iodide . One strain each of C . albicans and C . tropicalis (both sensitive to amphotericin B) and one strain of C . tropicalis (resistant to amphotericin B) were used to validate the method . Sensitivity or resistance was easily determined after 30 min of incubation in the presence of the antibiotic, and the drug effect was evident in a dose-dependent manner. Biochim Biophys Acta, 1995 Jun 14, 1236(2), 339 - 44 Horizontal 'solvent-free' lipid bimolecular membranes with two-sided access can be formed and facilitate ion channel reconstitution; Brutyan RA et al.; We present here an easily used method and apparatus for formation of horizontal 'solvent-free' lipid bilayer membranes affording two-sided access . These horizontal bilayers allow direct delivery of submicroliter volumes of samples onto the membrane upper surface increasing the efficacy of reconstitution by several orders of magnitude, as demonstrated using Staphylococcus aureus alpha-toxin . Also, they permit creation of locally high and transient transbilayer osmotic gradients to initiate fusion of ion-channel containing liposomes with planar membrane, which, following fusion, leaves the membrane and channel in essentially symmetric bathing solutions . This method is especially advantageous for cases where thickness of the membrane, absence of hydrocarbon solvent, or presence of differing lipid compositions in the two monolayers is critical. Arch Intern Med, 1995 Jun 12, 155(11), 1161 - 6 Staphylococcus aureus catheter-associated bacteremia . Minimal effective therapy and unusual infectious complications associated with arterial sheath catheters; Malanoski GJ et al.; OBJECTIVE: To determine factors that predict complications and examine outcomes of Staphylococcus aureus bacteremia according to the duration of antibiotic therapy . METHODS: Clinical data were extracted from charts of patients with positive blood cultures for S aureus at a single institution during a 2-year period . RESULTS: Of 102 patients with S aureus bacteremia, 55 were considered to have bacteremia attributable to an intravascular catheter, including five patients who were bacteremic after percutaneous transluminal coronary angioplasty . Among the other 50 patients with S aureus catheter-associated bacteremia, infection was community acquired in 18 and nosocomial in 32 . Septic pulmonary emboli were more common in patients with community-acquired S aureus catheter-associated bacteremia, most of whom had Hickman catheters or venous access disks . Delayed removal of the infected catheter was associated with persistence of bacteremia (P = .01) . With patients with early complications excluded, patients treated for 10 to 15 days had clinical characteristics similar to those of patients treated with longer courses of antibiotics and had similarly low rates of relapse (0% vs 4.7%) . In contrast, treatment with parenteral antibiotics for less than 10 days appeared to be inadequate in that relapse occurred in two of three such patients . Staphylococcus aureus catheter-associated bacteremia associated with percutaneous transluminal coronary angioplasty was complicated by a femoral artery mycotic aneurysm in two of five patients . CONCLUSION: Approximately one third of S aureus catheter-related bacteremias were community acquired, reflecting increased usage of intravascular devices for home parenteral support . A 10- to 15-day course of parenteral antibiotics was equivalent to longer courses of therapy in patients without early complications. Schweiz Med Wochenschr, 1995 Jun 10, 125(23), 1151 - 61 {Bacterial resistance to antibiotics}; Moreillon P; 50 years ago, the introduction of penicillin, followed by many other antibacterial agents, represented an often underestimated medical revolution . Indeed, until that time, bacterial infections were the prime cause of mortality, especially in children and elderly patients . The discovery of numerous new substances and their development on an industrial scale gave us the illusion that bacterial infections were all but vanquished . However, the widespread and sometimes uncontrolled use of these agents has led to the selection of bacteria resistant to practically all available antibiotics . Bacteria utilize three main resistance strategies: (1) modification of their permeability, (2) modification of target, and (3) modification of the antibiotic . Bacteria modify their permeability either by becoming impermeable to antibiotics, or by actively excreting the drug accumulated in the cell . As an alternative, they can modify the structure of the antibiotic's molecular target--usually an essential metabolic enzyme of the bacterium--and thus escape the drug's toxic effect . Lastly, they can produce enzymes capable of modifying and directly inactivating antibiotics . In addition, bacteria have evolved extremely efficient genetic transfer systems capable of exchanging and accumulating resistance genes . Some pathogens, such as methicillin-resistant Staphylococcus aureus and multiresistant Mycobacterium tuberculosis, have become resistant to almost all available antibiotics and there are only one or two substances still active against such organisms . Antibiotics are very precious drugs which must be administered to patients who need them . On the other hand, the development of resistance must be kept under control by a better comprehension of its mechanisms and modes of transmission and by abiding by the fundamental rules of anti-infectious chemotherapy, i.e.: (1) choose the most efficient antibiotic according to clinical and local epidemiological data, (2) target the bacteria according to the microbiological data at hand, and (3) administer the antibiotic in an adequate dose which will leave the pathogen no chance to develop resistance. J Biol Chem, 1995 Jun 2, 270(22), 13541 - 7 Ceramide triggers meiotic cell cycle progression in Xenopus oocytes . A potential mediator of progesterone-induced maturation; Strum JC et al.; The role of sphingomyelin-derived second messengers in progesterone-induced reinitiation of the meiotic cell cycle of Xenopus laevis oocytes was investigated . A brief treatment of defolliculated oocytes with sphingomyelinase (Staphylococcus aureus) was sufficient to induce maturation as measured by H1 kinase activity and germinal vesicle breakdown (GVBD) . Pretreatment with cycloheximide inhibited sphingomyelinase-induced GVBD demonstrating a requirement for protein synthesis . Microinjection of ceramide or sphingosine, potential products of sphingomyelin hydrolysis, were capable of inducing GVBD in the absence of hormone . Metabolic labeling studies suggested the conversion of sphingosine to ceramide was necessary for sphingosine-induced GVBD . Additionally, fumonisin b1, an inhibitor of sphingosine N-acyltransferase, blocked sphingosine-induced GVBD demonstrating that ceramide is the more proximal biologically active metabolite . Treatment of oocytes with progesterone, the physiological inducer of oocyte maturation, resulted in a time- and concentration-dependent increase in the mass of ceramide and decrease in the mass of sphingomyelin through activation of a Mg(2+)-dependent neutral sphingomyelinase . These observations suggest that the generation of ceramide from sphingomyelin is part of the signal transduction pathway activated in response to progesterone and that the increase in ceramide is likely to be functionally important in resumption of the meiotic cell cycle. Afr J Med Med Sci, 1995 Jun, 24(2), 135 - 8 Plasmid profiles and antibiotic susceptibility patterns of Staphylococcus aureus isolates from Nigeria; Olukoya DK et al.; In an investigation into the problems of infections due to Staphylococcus aureus in Nigeria, 100 strains were isolated from various hospitals in Lagos . The strains were screened for the presence of plasmids and for susceptibility to antimicrobial agents . Plasmids were extracted by modification of the method of Takahashi and Nagono{1} . The plasmids were diverse in nature . The strains were found to be highly resistant to commonly prescribed antibiotics. Q J Nucl Med, 1995 Jun, 39(2), 89 - 98 A new 99mTc labelling method for leucocytes: in vitro and in vivo comparison with 99mTc-HMPAO; Welling M et al.; A new method for the labelling of mixed leucocytes with 99mTc-tropolone was optimized and compared with a 99mTc-HMPAO leucocyte labelling procedure in vitro and in vivo . In the present study, leucocytes obtained from patients suffering from Crohn's disease, were isolated and labelled with 99mTc-HMPAO or labelled according the new 99mTc-tropolone procedure using 9.8 mM tropolone, 1 microM stannous chloride and 0.8 mM potassium borohydride (KBH4) at pH 5.5-6 . Labelling efficiency with 99mTc-tropolone yielded 92 +/- 3%, which is higher compared to the 99mTc-HMPAO labelling procedure (64 +/- 13%) using 10(8) of leucocytes . In vitro stability and viability of both the tropolone and the HMPAO labelled cells was investigated . The viability test of the 99mTc-labelled leucocytes was performed in autologous plasma at 37 degrees C and compared with unlabelled leucocytes . After 18 hours of incubation a significant (P < 0.05) higher stability was observed for 99mTc-tropolone labelled leucocytes (84 +/- 5%) compared with that of 99mTc-HMPAO labelled leucocytes (73 +/- 5%) . The viability of the 99mTc-labelled leucocytes observed for both labelling procedures was similar to unlabelled leucocytes . In vivo experiments were performed in mice . 99mTc-tropolone or 99mTc-HMPAO labelled murine mixed leucocytes were injected in mice, with a Staphylococcus aureus ATCC 25923 thigh infection . Analysis of scintigraphic images yielded a faster clearance of the 99mTc-tropolone labelled leucocytes . This was most likely due to a significant (P < 0.02) higher liver uptake at 4 hours after administration of the 99mTc-tropolone labelled leucocytes (19%) in comparison with 99mTc-HMPAO labelled cells (9%) . Faster and significant (P < 0.02) higher accumulation of the 99mTc-tropolone labelled leucocytes was observed at the site of infection compared with 99mTc-HMPAO labelled leucocytes at all time-intervals after the administration of the 99mTc-labelled leucocytes . The new 99mTc-tropolone leucocyte labelling procedure, offers an attractive low-cost agent for research purposes. Lett Appl Microbiol, 1995 Jun, 20(6), 391 - 5 Antimicrobial drug resistance in Staphylococcus aureus isolated from cattle in Brazil; Pereira MS et al.; Isolates of Staphylococcus aureus obtained from apparently healthy cattle in the State of Paraiba, Brazil were characterized in relation to resistance to 21 antimicrobial agents . Among the 46 isolates obtained, resistance to penicillin was most frequent, followed by resistance to cadmium, streptomycin, arsenate, tetracycline, mercury, erythromycin and kanamycin/neomycin . All isolates were susceptible to fusidic acid, ethidium bromide, cetrimide, chloramphenicol, benzalkonium chloride, doxycycline, gentamicin, methicillin, minocycline, novobiocin, rifamycin, tylosin and vancomycin . Only six isolates were susceptible to all the drugs tested . With respect to the antibiotics, multi-resistant isolates were uncommon . These results are probably a consequence of the peculiarities of local drug usage pressures . In relation to metal ions, resistance to mercury was rare while resistance to arsenate was relatively frequent, which contrasts with the situation for human Staph . aureus strains . After treatment with ethidium bromide, elimination of resistance to penicillin, tetracycline, streptomycin, erythromycin and cadmium was observed, which was consistent with the genetic determinants being plasmid-borne. FEMS Microbiol Lett, 1995 Jun 1, 129(1), 11 - 5 Intrinsic insensitivity to cadmium of the L-lactate oxidizing system in staphylococcoccus aureus; Tynecka Z et al.; Starved cells of cadmium-sensitive Staphylococcus aureus 17810S accumulated 109Cd via the Mn2+ porter energized by the membrane potential (delta psi) generated by L-lactate oxidation . However, Cd2+ accumulation did not result in inhibition of respiration and consequent generation of electrochemical proton gradient (delta muH+) via the respiratory chain . Thus, delta muH(+)-consuming processes, such as ATP synthesis and {14C}glutamate transport proceeded normally, despite the presence of Cd2+ in the cytoplasm . The mechanism of the intrinsic cadmium-insensitivity of the L-lactate oxidizing system is discussed. Chest, 1995 Jun, 107(6), 1522 - 5 The role of the 'pericardial window' in AIDS; Flum DR et al.; BACKGROUND: The leading cause of pericardial effusion in urban hospitals is now AIDS-related pathologies . Clinically, these effusions are a diagnostic and management dilemma . In our institution, pericardial biopsy and operative drainage have become part of the diagnostic and management plan . Surgical intervention, however, has appeared to have little clinical impact . METHODS: A retrospective review was conducted of all patients (n = 29) diagnosed as having AIDS who underwent "pericardial window" for pericardial effusions from 1986 to 1994 . RESULTS: Fluid cultures and pericardial biopsy were performed in all cases . Twenty-four percent of culture or biopsy specimens were diagnostic (7 of 29 = 2 adenocarcinoma, 3 lymphoma, 1 Staphylococcus aureus, 1 Mycobacterium tuberculosis) . In 94% of cases, there was no change in clinical management based on operative results . In 4 of 7 cases, the patients were ineligible for the indicated therapy based on underlying illness and in 1 of 7, the patient was receiving appropriate therapy for previously diagnosed disease . Ventilatory complications were noted in 17% . Three patients did not wean from the ventilator and died shortly after the operation . Sixty-nine percent mortality was noted at 8 weeks post-operatively . One hundred percent mortality was noted at 22 weeks with 86% follow-up . CONCLUSION: AIDS-related pericardial effusion is associated with a grave prognosis . Operations for diagnostic benefit provide little practical information and are not justified. Clin Exp Immunol, 1995 Jun, 100(3), 406 - 11 Two novel cationic staphylococcal proteins induce IL-2 secretion, proliferation and immunoglobulin synthesis in peripheral blood mononuclear cells (PBMC) of both healthy controls and patients with common variable immunodeficiency (CVID); Jahreis A et al.; Two cationic proteins, a neutral phosphatase (NP-tase) and a 70-kD protein (p70) were isolated from Staphylococcus aureus by ion exchange chromatography . We compared their properties to those of the well established B cell mitogen of whole, fixed Staph . aureus strain Cowan I cells (SAC) . Both purified proteins were able to induce immunoglobulin synthesis in PBMC cultures of healthy donors . NP-tase and p70 also induced immunoglobulin synthesis of PBMC from those patients with CVID who were also responsive to SAC plus IL-2 stimulation . Immunoglobulin synthesis in response to NP-tase and to p70 was time- and dose-dependent and could be inhibited by addition of specific antibodies against the proteins . In contrast to SAC, no addition of exogenous IL-2 was necessary to obtain maximal immunoglobulin synthesis induced by NP-tase or p70 . However, neither protein was able to induce immunoglobulin synthesis in B cell-enriched cultures . High amounts of IL-2 were found in supernatants of PBMC from healthy donors following stimulation with low concentrations of NP-tase or p70, and this was associated with vigorous lymphocyte proliferation . Both proteins behave like typical antigens, and not like lectins or superantigens, since an NP-tase-stimulated T cell line showed an antigen-specific, MHC-restricted secondary response . In addition, no preferential T cell receptor V beta chain usage was found with eight V beta-specific MoAb . It is likely that the two proteins induce antigen-specific T cell activation, which is then followed by polyclonal activation of B cells via CD40 receptors and cytokine release. J Infect Dis, 1995 Jun, 171(6), 1646 - 50 Decreased teicoplanin susceptibility of methicillin-resistant strains of Staphylococcus aureus; Mainardi JL et al.; Between February 1992 and February 1993, 12 patients were seen who were infected or colonized with methicillin-resistant strains of Staphylococcus aureus . The strains had decreased susceptibility to teicoplanin (MICs, 8-16 micrograms/mL) . Field inversion gel electrophoresis showed the strains belonged to three related clones (A1, A2, and A3) . The patients were though to have acquired the strains nosocomially . Consistent with results of laboratory studies of teicoplanin-resistant S . aureus, all but 1 strain expressed a 35-kDa membrane protein, and 9 strains expressed increased levels of penicillin-binding protein 2 complex . Six strains were isolated from patients treated with glycopeptides . These data suggest that nosocomial transmission of S . aureus with decreased teicoplanin susceptibility may occur during glycopeptide use and that such strains develop resistance by a mechanism associated with the appearance of a 35-kDa membrane protein . Surveillance is necessary to monitor for the potential selection of resistant clones that may be capable of dissemination. J Bacteriol, 1995 Jun, 177(11), 2965 - 70 Characterization of the gene for the chromosomal dihydrofolate reductase (DHFR) of Staphylococcus epidermidis ATCC 14990: the origin of the trimethoprim-resistant S1 DHFR from Staphylococcus aureus? Dale GE, Broger C, Hartman PG, Langen H, Page MG, Then RL, Stuber D. The gene for the chromosomally encoded dihydrofolate reductase (DHFR) of Staphylococcus epidermidis ATCC 14990 has been cloned and characterized . The structural gene encodes a polypeptide of 161 amino acid residues with a calculated molecular weight of 18,417 . This trimethoprim-sensitive (Tmps) DHFR, SeDHFR, differs in only three amino acids (Val-31-->Ile, Gly-43-->Ala, and Phe-98-->Tyr) from the trimethoprim-resistant (Tmpr) S1 DHFR encoded by transposon Tn4003 . Since in addition the S . epidermidis gene also forms part of an operon with thyE and open reading frame 140 as in Tn4003, the chromosomally located gene encoding the Tmps SeDHFR is likely to be the molecular origin of the plasmid-located gene encoding the Tmpr S1 DHFR . Site-directed mutagenesis and kinetic analysis of the purified enzymes suggest that a single Phe-->Tyr change at position 98 is the major determinant of trimethoprim resistance. Infect Immun, 1995 Jun, 63(6), 2141 - 6 A toxic shock syndrome toxin 1 mutant that defines a functional site critical for T-cell activation; Cullen CM et al.; Toxic shock syndrome toxin 1 (TSST-1), a superantigen produced by Staphylococcus aureus, is a causative agent of toxic shock syndrome (TSS) . This superantigen is a potent stimulator of T cells and macrophages/monocytes, resulting in the release of cytokines that are implicated in the pathogenesis of TSS . This study characterizes a mutant TSST-1, derived by site-directed mutagenesis, that has an alanine substitution at histidine 135 (mutant 135) . This single-amino-acid change results in a mutant toxin that has lost mitogenic activity for T cells . In contrast to wild-type TSST-1, this mutant does not induce T cells to express interleukin-2, gamma interferon, or tumor necrosis factor beta (TNF-beta) . The inability of mutant 135 to activate T cells is not due to a lack of binding to the class II major histocompatibility complex receptor . In addition, the mutant TSST-1 does not induce expression of TNF-alpha, which plays a role in the development of lethal shock . The lack of TNF-alpha induction by mutant 135 is likely due to its inability to activate T cells . These data suggest that the mutation at histidine 135 in TSST-1 affects toxin interactions with the T-cell receptor rather than the class II major histocompatibility complex receptor. J Immunol, 1995 Jun 1, 154(11), 6174 - 81 Down-regulating effects of IL-4 and IL-10 on the IFN-gamma response in atopic dermatitis; Lester MR et al.; Atopic dermatitis (AD) is a chronic allergic disease associated with toxin (superantigen)-producing Staphylococcus aureus skin infections, impaired delayed hypersensitivity responses, and the expansion of IL-4-secreting Th2 cells, as well as diminished IFN-gamma synthesis . IL-12 is known to induce IFN-gamma synthesis and to augment Th1 responses . In this study, therefore, we examined the potential role of IL-12 in the immunopathogenesis of AD . We show that, after stimulation with staphylococcal toxic shock syndrome toxin-1 (TSST-1) or IL-12, PBMC from patients with AD are deficient in their ability to produce IFN-gamma . PBMC from AD patients, however, produced normal quantities of IL-12 and expressed normal levels of IL-12R . Induction of IFN-gamma by TSST-1 was decreased by neutralizing anti-IL-12 Ab in normal donors, but not in AD patients . The latter observation is consistent with a defective response to IL-12 in AD PBMC . Because AD is associated with increased production of IL-4 and IL-10, we examined the effect of IL-4 on IL-12- or TSST-1-induced IFN-gamma production in normal donors . IL-4 inhibited IL-12-induced IFN-gamma production . Furthermore, Ab neutralization of IL-4 caused increased production of IFN-gamma in AD PBMC . However, neutralization of IL-10 activity caused an even greater augmentation of IFN-gamma production . Our data suggest that despite normal levels of IL-12 production and IL-12R expression, PBMC from AD patients are unable to generate normal IL-12-induced IFN-gamma responses . This defective response may be due to the excess production of IL-4 and IL-10 in this common allergic condition. Am J Infect Control, 1995 Jun, 23(3), 200 - 8 Use of 0.3% triclosan (Bacti-Stat) to eradicate an outbreak of methicillin-resistant Staphylococcus aureus in a neonatal nursery; Zafar AB et al.; BACKGROUND: Once established in an institution, methicillin-resistant Staphylococcus aureus (MRSA) outbreaks have proved difficult to eradicate, despite intensive infection control measures . This report describes the nosocomial infection with MRSA of 22 male infants in a neonatal nursery during a 7-month period and the infection control procedures that effectively brought this outbreak under control and eliminated recurrence for more than 3 1/2 years . METHODS: After a single index case of bullous impetigo caused by MRSA in a neonate discharged from the nursery 2 weeks previously, an additional 18 cases of MRSA skin infections were clustered in a 7-week period . Aggressive infection control measures were instituted, including changes in umbilical cord care, circumcision procedures, diapers, handwashing, gloves, gowns, linens, disinfection, placement in cohorts of neonates and staff, surveillance, and monitoring . RESULTS: These measures were not effective in slowing the outbreak . The single additional measure of changing handwashing and bathing soap to a preparation containing 0.3% triclosan (Bacti-Stat) was associated with the immediate termination of the acute phase of the MRSA outbreak . CONCLUSION: The nursery has remained free of MRSA for more than 3 1/2 years, attesting to the success of our program. Am J Infect Control, 1995 Jun, 23(3), 194 - 9 Long-term central venous catheters in patients with acquired immunodeficiency syndrome; Sweed M et al.; BACKGROUND: As long-term vascular access becomes more prevalent among patients with AIDS, it is becoming more important to consider their potential complications . METHODS: One hundred two central venous access devices placed in 84 patients with AIDS were reviewed for septic and mechanical complications . Catheters were inserted by one surgeon by means of the cephalic vein cutdown technique . The sample included 88 implanted venous reservoir catheters (86.3%) and 14 tunneled central venous catheters (13.7%) . RESULTS: Mean catheter life was 141 +/- 15 days . Total number of catheter days was 14,383 . The catheter-related infection rate was 0.125 episodes/100 catheter-days . Staphylococcus aureus was the most commonly isolated pathogen in the sample . Mechanical complications were rare (0.05 episodes/100 catheter-days) . CONCLUSION: When these data are compared with other, smaller series in the literature, the findings suggest that long-term central venous catheters inserted in patients with AIDS are safe and effective for the multiple infusion therapies required in these patients. J Otolaryngol, 1995 Jun, 24(3), 206 - 8 Bacteriology of chronic suppurative otitis media: ofloxacin susceptibility; Yuen AP et al.; Fifty-four patients with active chronic suppurative otitis media (CSOM) were prospectively studied for the bacteriology and their in vitro antibiotic susceptibility to ofloxacin . Thirty-nine patients (72%) had positive cultures . The commonest organism was Pseudomonas sp., which was found in 18 (33%) patients . The second commonest organism was Staphylococcus aureus, which was found in 15 (28%) patients . Bacteroides sp . was found only in 3 (6%) patients, and all were in association with the aerobes . Of all 42 aerobes found, 35 (83%) were susceptible to ofloxacin . The in vitro susceptibility results indicate that ofloxacin can be an effective antibiotic in the treatment of active CSOM. J Hosp Infect, 1995 Jun, 30(2), 111 - 24 beta-Lactamase production and genetic location in Staphylococcus aureus: introduction of a beta-lactamase plasmid in strains of phage group II; Skov RL et al.; Staphylococcus aureus strains of phage group II have increased in frequency in hospital-acquired infections during recent years . A total of 184 penicillin resistant group II strains from bacteraemia cases in the years 1961-1990 were analysed for the amount of beta-lactamase produced and the location of the beta-lactamase gene . Until 1977 all strains investigated had a chromosomally located beta-lactamase gene, but since then a 21 kb beta-lactamase plasmid has increased in occurrence among group II strains, especially among those strains typable only at high phage concentrations {100 x Routine Test Dilution (RTD) and 1000 x RTD} . In 1990, 84% of the group II strains contained this plasmid . Plasmid-containing strains produced more beta-lactamase than strains without the plasmid . S . aureus strains of the 94,96 complex, which since 1984 have decreased in frequency from 18 to 9% in 1993, have remained high producers of beta-lactamase. Nihon Rinsho Meneki Gakkai Kaishi, 1995 Jun, 18(3), 265 - 71 {Modulation of immunoglobulin production by polymorphonuclear neutrophils}; Yoshino Y; Infiltration of polymorphonuclear neutrophils (PMN) as well as lymphocytes is commonly observed at the sites of inflammation . However, it is still unclear whether PMN influence the function of lymphocytes . The current studies examined the effects of PMN upon immunoglobulin production . PMN did not significantly influence the IgM production by B cells stimulated with Staphylococcus aureus (SA) + IL-2 in the absence of T cells . By contrast, PMN significantly enhanced the IgM production induced by immobilized anti-CD 3 stimulated T cells . PMN also enhanced the interferon-gamma (IFN-gamma) production by immobilized anti-CD 3 stimulated T cells . PMN fixed by paraformaldehyde enhanced the IgM production and the IFN-gamma production induced by immobilized anti-CD3 as effectively as fresh PMN . These results indicate that PMN enhance the Ig production by upregulating the functions of immobilized anti-CD 3-activated T cells through direct cellular interaction with T cells. Semin Arthritis Rheum, 1995 Jun, 24(6), 391 - 410 Septic bursitis; Zimmermann B 3rd et al.; Nine cases of septic bursitis are presented, and the literature on the subject comprehensively reviewed, with an emphasis on the clinical manifestations of septic bursitis in various anatomic locations . Physical activities associated with increased susceptibility to septic bursitis and systemic conditions that increase the severity of septic bursitis are catalogued . Analysis of the microbiology of cases reported in the literature demonstrates that greater than 80% of cases of septic bursitis are caused by Staphylococcus aureus and other gram-positive organisms . However, a wide variety of gram-negative microorganisms, fungi, and other infectious agents have been reported to cause septic bursitis and may lead to complications in diagnosis and treatment . The nine cases reported here demonstrate the potential severity of septic bursitis and emphasize that significant systemic complications may result from this common musculoskeletal infection . Indications for hospitalization and/or intravenous antibiotic therapy for septic bursitis include the presence of fulminant local infection, evidence for systemic toxicity, or infection in an immunocompromised patient . Patients who fail to respond to intravenous antibiotics and percutaneous aspiration of the bursa may require surgical drainage or bursectomy by one of several methods that have been proposed . There is some recent evidence that intrabursal corticosteroid injection for therapy of nonseptic subcutaneous bursitis may be more effective than systemic antiinflammatory medication or simple bursa aspiration. Int J Food Microbiol, 1995 Jun, 26(1), 15 - 24 Media used in the detection and enumeration of Staphylococcus aureus; Baird RM et al.; Over the years a range of selective and diagnostic media have been developed to assist in the detection and enumeration of Staphylococcus aureus in routine food surveillance programmes and food poisoning investigations and these are reviewed here . Baird-Parker agar remains, however, the medium of choice for direct plating and enumeration of S . aureus in both Europe and the US . This paper also reports on a comparison of the productivity and selectivity of trypticase soy broth with 10% NaCl and 1% sodium pyruvate (PTSBS) with trypticase soy broth (TSB) for the isolation of S . aureus . Using three strains of S . aureus and a strain of S . hyicus the productivity ratio of PTSBS to TSBS ranged from -0.17 to 0.57 . In the recovery of heat-injured cells of S . aureus, PTSBS offered little or no improvement over TSB with sodium chloride but no pyruvate in a three-tube MPN assessment . Of the seven other bacterial cultures tested, none grew well on the PTSBS . Selectivity ratios of 4.4-7 were found. AIDS, 1995 Jun, 9(6), 547 - 53 Lack of selective V beta deletion in CD4+ or CD8+ T lymphocytes and functional integrity of T-cell repertoire during acute HIV syndrome; Cossarizza A et al.; OBJECTIVE: To study the V beta T-cell repertoire in peripheral blood lymphocytes (PBL) during acute HIV syndrome by using several anti-V beta monoclonal antibodies (MAb) and to analyse its functionality by stimulating PBL with superantigens (SAg) such as Staphylococcus aureus enterotoxins . METHODS: Cytofluorimetric analysis of V beta T-cell-receptor expression was performed on PBL from eight patients with symptomatic, acute HIV-1 primary infection, showing a dramatic decrease of CD4+ PBL accompanied by a marked increase in activated/memory CD8+ T cells, and on 12 age- and sex-matched healthy controls . PBL were then isolated, stimulated with different SAg, anti-CD3 MAb or phytohaemagglutinin and cultured for 3 days . PBL capability to progress through cell cycle was studied by the classic cytofluorimetric method of bromodeoxyuridine incorporation and DNA staining with propidium iodide . RESULTS: Despite the presence of a few expansions of some V beta families among CD8+ T lymphocytes, no gross alterations in T-cell repertoire were present in patients with acute HIV syndrome . Its functionality was maintained overall, as PBL responsiveness to SAg was well preserved . Interestingly, all CD8+ T cells, although bearing different V beta T-cell receptors, expressed marked signs of activation, i.e., CD45R0, CD38 and major histocompatibility complex class II molecules, and also high amounts of CD11a and CD18 . CONCLUSIONS: Our data suggest, at least in the early phases and in the acute form of the infection, that HIV is not likely to act as a SAg . However, further studies are needed to analyse other sites, such as lymph nodes, where HIV could exert other, significant effects, and to study the expression of other V beta families than those investigated here. Infect Control Hosp Epidemiol, 1995 Jun, 16(6), 354 - 8 Mupirocin resistance: clinical and molecular epidemiology; Bradley SF et al.; The antistaphylococcal activity of topical mupirocin has made it an attractive agent for the treatment of asymptomatic colonization with Staphylococcus aureus . Increasing use has been associated with the emergence of mupirocin resistance in staphyloccoci, and failure of therapy has been associated with the isolation of strains exhibiting high-level resistance (MIC > 500 micrograms/mL) . Fortunately, low-level mupirocin resistance (MIC < 100 micrograms/mL) occurs most commonly . Because a novel gene encoding for mupirocin resistance resides in both low-level and high-level resistant strains, the emergence of low-level mupirocin resistance may not be as epidemiologically insignificant as previously thought. J Reprod Med, 1995 Jun, 40(6), 485 - 6 Amnionitis and premature delivery with intact amniotic membranes involving Staphylococcus aureus . A case report; Ben-David Y et al.; Subclinical infection is suspected to be an important etiologic factor in the initiation of preterm labor in women with intact membranes . We present a case of acute clinical chorioamnionitis followed by preterm labor and fetal distress in a woman with intact membranes . The bacteriologic data on the mother and neonate clearly identified coagulase-positive Staphylococcus aureus as the etiologic factor. J Gastroenterol, 1995 Jun, 30(3), 408 - 12 A case of gallbladder cancer associated with a common bile duct neuroma, and a cystic lesion of the liver with histologic findings similar to those of an inflammatory pseudotumor; Akiyama T et al.; We report a rare case of gallbladder cancer associated with a common bile duct neuroma, and a cystic liver lesion with histologic findings similar to an inflammatory pseudotumor, in a patient who had had no previous abdominal surgery . The patient was a 62-year-old man whose major complaint was fever . Ultrasonography and a computed tomography scan revealed gallstones, an elevated lesion in the gallbladder, and a cystic liver lesion . Endoscopic retrograde cholangiopancreatography demonstrated stenosis of the common bile duct . Cultures of the cystic fluid and gallbladder bile were positive for Staphylococcus aureus . The patient underwent hepatectomy (inferior S4, S5, and S6), cholecystectomy, resection of the common bile duct, and right hemicolectomy . The resected specimens revealed gallbladder cancer with the microscopic appearance of a papillary adenocarcinoma, and a 12 x 4.5 x 3.5 cm cystic liver lesion with a wall 7 mm thick . Histologic studies of the wall of the cystic liver lesion revealed infiltration by histiocytes and plasma cells, and the presence of fibrous connective tissue, which findings are characteristic of inflammatory pseudotumors . A 9 x 6 mm elevated lesion, with the microscopic appearance of a neuroma, was resected from the common bile duct. Unfallchirurg, 1995 Jun, 98(6), 338 - 43 {Results of long-term therapy of chronic, post-traumatic osteomyelitis with gentamycin PMMA chains}; Jerosch J et al.; From 1977 to 1983, 173 patients suffering from chronic posttraumatic osteomyelitis were treated by local debridement and implantation of septopal beads; 102 patients were subsequently evaluated after follow-up periods of between 4 and 10 years . In 89.2% of these cases cure was achieved during the initial hospital stay . In 15.7% further debridements were necessary during the initial hospital stay to manage infection . In 91 patients there was no early recurrence until 1983 . After 1983 only 7 patients had late recurrences . At the time of follow up 95 patients (94.5%) did not show any signs of infection . It was necessary for 9 patients to change their profession because of the disease . Full use of the extremity involved was reported by 45 patients, and some slight limitations by 46 . In 11 cases the patients could not use the extremity involved in activities of daily living . In mono-infections (87.2%) and in infection with multiple different pathogen chains (82%) Staphylococcus aureus was by far the most common pathogen . Our findings indicate that both the outcome and the prognosis score are significantly better after the treatment regimen used than after other treatment modalities. Mol Immunol, 1995 Jun, 32(9), 603 - 12 In vitro induction of primary, antigen-specific CTL from human peripheral blood mononuclear cells stimulated with synthetic peptides; Wentworth PA et al.; A protocol for in vitro induction of primary, antigen-specific CTL from human peripheral blood mononuclear cells (PBMCs) was developed . Antigen presenting cells (APCs) consisted of Staphylococcus aureus Cowan-I (SAC-I) activated PBMCs treated with a citrate-phosphate buffer at pH 3 to release endogenous peptides bound to surface MHC . This treatment resulted in transient expression of empty class I molecules which could be subsequently stabilized with peptide and beta 2-microglobulin (beta 2m) . SAC-I activated PBMCs from HLA-A2.1 normal donors loaded with HBV core 18-27 peptide following acid treatment were used to stimulate PBMCs depleted of CD4+ T cells, in the presence of recombinant interleukin-7 (rIL-7) . After 12 days, cells were restimulated with autologous, peptide-pulsed, adherent cells and tested for CTL activity 7 days later . In 23 independent experiments from 13 different HLA-A2.1 donors, this protocol resulted in induction of primary CTL more than 90% of the time . As indicated by both the frequency and magnitude of the response against peptide-sensitized target cells, SAC-I activated PBMCs treated with acid were the most efficient stimulator APC . Thirteen per cent of the cultures generated were capable of lysing target cells transfected with the HBV core antigen and, in general, these CTL cultures exhibited high avidity for the HBV core peptide . This protocol is generally applicable to different antigens and class I alleles, and thus, may be utilized to screen large numbers of peptides to identify human CTL epitopes. Clin Orthop, 1995 Jun, (315), 153 - 62 Diagnosis and management of infection after tibial intramedullary nailing; Zych GA et al.; A series of 20 patients with infection after intramedullary nailing of the tibia is discussed . The most common pathogen was Staphylococcus aureus, which was found in 14 patients (64%) . Eleven nails were originally inserted without reaming, and 9 were reamed . Treatment protocols were based on the time of onset of infection (acute, subacute, and chronic) and the status of bone healing . In eight patients, the fractures (6) and nonunions (2) were healed at diagnosis of infection and were treated by debridement, nail removal, and antibiotics . Twelve patients had fractures (8) and nonunions (4) that were not healed . Four were treated with debridement, nail removal, and external fixation, and four with debridement and nail retention . The overall success rate for eradicating infection was 90% . Infection after unreamed nailing had fewer complications and a higher success rate for infection control than did reamed nailing . Risk factors identified in this study for infection are previous external fixation, severe open fracture, and substance abuse. AORN J, 1995 Jun, 61(6), 1023 - 7, 1030-4 Surgical patients with multiantibiotic-resistant bacteria; Ronk LL; Although antibiotics can cure most bacterial infections, there is an increasing number of bacteria that are resistant to antibiotics . Methicillin-resistant Staphylococcus aureus (MRSA) is becoming increasingly prevalent in US health care facilities . The majority of these infections are found in patients who have extensive burns or surgical wounds . As a result, perioperative nurses must be knowledgeable about MRSA and its implications for the OR . There are many theories on how to control the spread of MRSA but not one definitive set of control measures . Perioperative nurses, in cooperation with infection control practitioners, must develop policies that detail how patients with MRSA will be treated. Nippon Kyobu Geka Gakkai Zasshi, 1995 Jun, 43(6), 857 - 60 {A case of tricuspid valve endocarditis in the drug addict}; Hattori T et al.; We reported a case of tricuspid valve endocarditis in a drug addict . A 25-year-old woman with a history of drug abuse was admitted to our hospital for high fever . Blood cultures revealed staphylococcus aureus, and echocardiography showed vegetation attached to the tricuspid valve and moderate tricuspid regurgitation . Excision of the posterior leaflet including vegetation, and direct suture of the residual posterior leaflet along with annuloplasty was successfully done . Postoperative course was uneventful and endocarditis was eradicated. Kansenshogaku Zasshi, 1995 Jun, 69(6), 729 - 37 {Non-destructive and rapid identification of bacteria using near-infrared spectroscopy}; Matsunaga T et al.; Near-infrared (NIR) spectroscopy has recently come to be applied extensively in agricultural, food and chemical industries, and pharmaceutical science . We have been attempting to expand this method in the field of medical science . For example, we tried to use NIR spectroscopy for determination of bacteria . As the first step of this attempt, we differentiated between Escherichia coli and Staphylococcus aureus using NIR spectroscopy . This method could still further differentiate Methicillin-resistant Staphylococcus aureus (MRSA) and Methicillin-sensitive Staphylococcus aureus (MSSA) . Using those results as reference, the true name of bacteria from unknown bacteria was given . Not only untreated bacteria, but also we differentiated untreated MSSA, MSSA cultured in sub MIC concentration of ABPC and heat-killed MSSA . This identification method is sensitive to the bacterial concentration . In the future, the some new idea of a new direction of research from the result of plots of weights from two different bacteria will appear. Rinsho Byori, 1995 Jun, 43(6), 547 - 56 {Immunological investigations on pathogenesis of staphylococcal scalded skin syndrome}; Machida K; Staphylococcal exfoliative toxin (ET) is an extracellular product of Staphylococcus aureus isolated from patients with staphylococcal scalded skin syndrome (SSSS) which includes Ritter's disease, bullous impetigo and staphylococcal scarlet fever, and has been regarded as the causative agent of SSSS . The ET has not only a splitting effect at the granular layer of skin in human and mice but also an immunogenicity to human and mice . Using experimental animals and clinical specimens of patients with or without SSSS, the immunological investigation were performed and following results were obtained . 1) When some inbred and congenic resistant strains of mice were immunized with serotype A ET (ETA), they were divided into the high anti-ETA antibody producers (high responders) and the low responders . The gene controlling antibody response to ETA in mice is located in the I-A subregion in the major histocompatible complex (H-2 complex), and its function seems to be at least related to antigen recognition at the T-lymphocyte level . 2) Neonatal mice are generally susceptible to ETA regardless of their H-2 haplo-type . However, the neonatal mice born to a high-responder mother immunized with ETA were resistant to the subcutaneous challenge of ETA, but those born to an immunized low-responder mother were susceptible to the challenge . 3) The relationship between susceptibility and immune response to ETA in some mammalians could be divided into three groups: the possession of resistant skin and high production of antibody to ETA (rabbits and rats); the possession of resistant skin and low production of antibody to ETA (golden hamsters and guinea pigs); the possession of sensitive skin and various titers of antibody to ETA (humans and mice) . 4) The incidence of ET producing strains of Staphylococcus aureus in various clinical specimens obtained from patients without SSSS was 12% (50 out of 418 strains) in our epidemiological investigation . The percentages of antibody to ETA in sera obtained from healthy males and females were 23% and 29%, respectively . Eventually, ET-producing strains of Staphylococcus aureus seem to be spread out wider than the microbiological result obtained from clinical specimens . 5) The mitogenic responses of lymphocytes isolated from patients with the first impetigo could be measured by using PHA, and the five-of them were in the low range, whereas lymphocytes from patients with recurrent impetigo were in the normal range on this survey . Maybe this results suggest the T lymphocyte functions of some patients with impetigo are deteriorated.(ABSTRACT TRUNCATED AT 400 WORDS) J Trauma, 1995 Jun, 38(6), 958 - 9 Development of an epidural abscess following staphylococcal septicemia in an acutely burned patient: case report; Still JM et al.; A 57-year-old black female burned in a house fire sustained 22.5% total body surface area burns . On postburn day 45, she developed Staphylococcus aureus septicemia, and vancomycin was begun . On postburn day 50, quadriplegia developed . Magnetic resonance imaging revealed an anterior cervical mass, and cervical drainage of a staphylococcal cervical abscess was conducted . Antibiotics were continued . The patient regained almost complete neurologic function and was discharged doing well. Br J Pharmacol, 1995 Jun, 115(3), 522 - 6 All-or-none augmentation of Ca2+ sensitivity in alpha-toxin-permeabilized single smooth muscle cells from guinea-pig taenia caecum; Mita M et al.; 1 . Isolated smooth muscle cells from guinea-pig taenia caecum were permeabilized by use of Staphylococcus aureus alpha-toxin, and the sarcoplasmic reticulum Ca2+ store was depleted by exposure to 0.1 microM A23187 . 2 . Shortening of alpha-toxin-permeabilized single smooth muscle cells was induced by increasing free Ca2+ but was not induced by 0.2 microM free Ca2+ . 3 . Shortening of the permeabilized cells was caused by application of acetylcholine (ACh) with free Ca2+ concentration held at 0.2 microM . Permeabilized smooth muscle cells responded to 0.3 microM or 1 microM ACh with 0.2 microM Ca2+ with maximal shortening . The concentration-response relationship to ACh had a very steep slope and the cell shortening appeared to be an all-or-none response rather than a graded response, as was the shortening of intact cells to ACh . 4 . The shortening of permeabilized cells was also induced by application of guanosine 5'-triphosphate (GTP) with 0.2 microM free Ca2+, showing an all-or-none response . The threshold concentration of GTP that induced an all-or-none response was between 10 microM and 30 microM . 5 . These results suggest that Ca2+ sensitivity is augmented by stimulation of the muscarinic receptor or GTP-binding protein(s) in an all-or-none manner . It seems probable that this contributes to the all-or-none response to ACh in intact smooth muscle cells. Biomaterials, 1995 Jun, 16(9), 685 - 90 Disposable contact lenses and bacterial adhesion . In vitro comparison between ionic/high-water-content and non-ionic/low-water-content lenses; Arciola CR et al.; An in vitro quantitative study of the adhesion of a Staphylococcus aureus strain to two types of disposable contact lenses has been carried out . The first type was an ionic/high-water-content (I-HWC) lens (42% Etafilcon A, 58% water) and the second was a non-ionic/low-water-content (Nl-LWC) lens (61.4% poly(2-hydroxyethyl methacrylate), 38.6% water) . Adhesion to the two lens types was evaluated both in basic conditions and after treatment with lysozyme . The results showed that I-HWC lenses are more prone to Staphylococcus aureus adhesion than NI-LWC lenses, both untreated (+15.4%) and treated with lysozyme (+20.5%) . Lysozyme increased bacterial adhesion by 30.5% on the lenses with lower water content, and by 36.3% on those with higher water content. Biomaterials, 1995 Jun, 16(9), 681 - 4 Seven surgical silicones retain Staphylococcus aureus differently in vitro; Arciola CR et al.; In an in vitro study, we have quantitatively evaluated the capability of seven different types of silicone to retain a Staphylococcus aureus strain, isolated from a surgical wound . All the silicone specimens were taken from prostheses already used in plastic or ophthalmological surgery . Two polymers were used as controls: polystyrene, because of its known capability to favour in vitro bacterial recovery, and nylon, for its bacterial repellence . The results show that all silicones are suitable substrata for Staphylococcus aureus . However, there are some differences among silicone types . The amounts of bacteria retained from silicone oils are greater than or equal to those obtained from the positive control material. J Hosp Infect, 1995 Jun, 30 Suppl, 465 - 71 New threats to the control of methicillin-resistant Staphylococcus aureus; Casewell MW; Several countries have achieved considerable success in the control of epidemic methicillin-resistant Staphylococcus aureus (MRSA) . However, in several hospitals in the UK, MRSA strains of enhanced epidemicity, notably EMRSA-16, are becoming endemic . Our inability to eliminate the cause of a single-strain outbreak is unfamiliar and unnerving . Factors in 'market-led' health care delivery that hinder control of MRSA include a shortage of inpatient beds, patients moving from ward to ward, and more mixed-specialty wards . Increasing use of day treatments leaves an inpatient hospital population with more risk factors for infection . Early discharge of infected patients to convalescent homes, or to homes for the elderly, has created a new reservoir of infected and colonized patients . The emergence of high-level mupirocin resistance may soon also contribute to failure of control . The transfer of vancomycin resistance from Enterococcus faecium to a laboratory strain of S . aureus suggests that, especially in hospitals with both vancomycin-resistant enterococci and MRSA, there is the opportunity for the emergence of vancomycin-resistant MRSA for which there may be no effective antimicrobial prophylaxis or treatment . It is increasingly important to persuade hospital managers that even partial control of MRSA, whilst expensive, is still cost-effective and is a quality issue for individual hospitals . The control of EMRSA-16 in one hospital has recently been estimated to have saved more than 629,000 pounds extra costs . MRSA continues to be at the forefront of those organisms that seriously challenge modern technological medicine and surgery. J Hosp Infect, 1995 Jun, 30 Suppl, 389 - 96 A review of the test methods used to establish virucidal activity; Bellamy K; Testing the efficacy of disinfectants has been the domain of bacteriologists for many years . Recently interest has grown in the virucidal effects of disinfectants, due to increased awareness of viral infections and concern for possible cross-infection . Findings have demonstrated significant differences in the susceptibility of certain viruses, particularly non-enveloped viruses, e.g . enteroviruses, to disinfectants compared to some bacteria . For example Escherichia coli and herpes simplex virus are inactivated by 20% isopropyl alcohol (IPA) whereas Staphylococcus aureus requires > or = 50% and poliovirus is not inactivated by IPA . Currently there is little or no standardization in the methods used for the determination of virucidal activity in suspension, or on hands and surfaces . Methods in use in Europe and the USA will be reviewed and their relevance to the clinical situation discussed. J Hosp Infect, 1995 Jun, 30 Suppl, 26 - 34 Infection control programmes--are they cost-effective? Mehtar S. Infection control (IC) programmes are cost-effective in the long-term but much depends on the available resources and the support from management . The funding of IC programmes at present is linked to the Microbiology Department and a separate budget needs to be established . The best use of resources is to apply a risk assessment to each situation which presents and to adapt protocols accordingly . For example, the treatment of a carrier or an infected patient with methicillin-resistant Staphylococcus aureus cost 374 pounds and 2454 pounds, respectively in 1993, the major portion of the cost being due to an increased length of stay which was two days and 10 days, respectively . It is more cost-effective to treat carriers . The other cost-effective investment is in education and reinforcement of simple messages . Formal lectures seem to be the least effective way of producing long-term effect; frequent ward visits or contacts are most effective . Also, there is better compliance when there is a perceived risk to the staff themselves . The availability of the IC team to advise helps reduce waste and therefore cost . This is particularly true of antibiotic usage where it was noted that without guidance, the antibiotic usage increased by 2000 pounds per month when compared to a similar period in the previous year . The available provisions for IC programmes in the UK are utilized exceptionally well when compared with other countries. J Antimicrob Chemother, 1995 Jun, 35(6), 865 - 8 Activity of the semi-synthetic kanamycin B derivative, arbekacin against methicillin-resistant Staphylococcus aureus; Hamilton-Miller JM et al.; Arbekacin (1-N-((S)-4-amino-2-hydroxybutyryl)-3',4'-dideoxykanamycin B) was active against 54 strains of methicillin-resistant Staphylococcus aureus from 16 different countries, all of which were resistant to > or = 32 mg/L amikacin . MICs of arbekacin ranged from 1 to 16 mg/L, the MIC50 was 3.2 mg/L and the mode geometric mean MICs were 4 and 4.5 mg/L respectively. Int Dent J, 1995 Jun, 45(3), 218 - 22 The effect of the extract of the miswak (chewing sticks) used in Jordan and the Middle East on oral bacteria; Al lafi T et al.; Chewing sticks are commonly used in Jordan, Saudi Arabia and the United Arab Emirates in particular, and the Middle East, Asia, and Africa in general, in addition to many other areas for oral hygiene, religious and social purposes . Recently, the World Health Organisation (WHO) has recommended and encouraged the use of these sticks as an effective tool for oral hygiene . The antibacterial activity of one of these sticks has been tested against some oral aerobic and anaerobic bacteria . Three methods of antibacterial activity were carried out: streaked plate method, ditch plate method, and tube dilution test for minimum inhibitory concentration (MIC) . It was found that the extract of these sticks had a drastic effect on the growth of Staphylococcus aureus with MIC values of 69 mg/100 cc, while a variable effect on other bacterial species was noted . It is concluded that using chewing sticks twice a day on a regular basis may reduce the incidence of gingivitis and possibly dental caries . Apart from their antibacterial activity which may help control the formation and activity of dental plaque, they can be used effectively as a natural toothbrush for teeth cleaning . Such sticks are effective, inexpensive, common, available, and contain many medical properties. Zhongguo Zhong Xi Yi Jie He Za Zhi, 1995 Jun, 15(6), 347 - 50 {Experimental and clinical study of shuanghuanglian aerosol in treating acute respiratory tract infection}; Wang YH et al.; Two hundrde and two cases of acute respiratory tract infection (ARI) were treated with Shuanghuanglian (SHL) aerosol . In these cases, the majority of cases were virus infection and 64% of them was caused by the respiratory syncytial virus (RSV) . The virostatic and bacteriostatic test were done in vitro by the cell culture method and it was shown that SHL could inhibit the RSV, para-influenza I-IV and 23 kinds of pathogenic bacteria such as Staphylococcus aureus etc . The bacteriostatic effect was positively correlated to the SHL concentration . Experimental study showed that SHL could enhance the NK cell activity, promote the production of alpha-interferon and raise the rate of lymphocyte transformation . The controlled observation on SHL preparation with various dosage-forms revealed that the SHL aerosol in treating early ARI showed better results than that of injections and oral liquor symptomatologically (P < 0.01) . The effective rate was 96%. J Vet Med Sci, 1995 Jun, 57(3), 581 - 2 Protein A in Staphylococcus aureus isolates from pigs; Takeuchi S et al.; The presence and quantity of protein A in Staphylococcus aureus 147 isolates from the tonsils of healthy pigs were examined by three methods . Cell-bound protein A was detected in 71 (48%), 104 (71%) and 123 (84%) of 147 isolates by the slide hemagglutination test, microplate hemagglutination test and enzyme-linked immunosorbent assay (ELISA), respectively . Extracellular protein A was not detected in any isolates by the microplate hemagglutination test . When the quantity of cell-bound protein A in the isolates was determined by the ELISA, most of the isolates contained about 0.8 to 2.2 micrograms of protein A/ml in bacterial cell suspensions of a concentration of MacFarland No . 3. J Vet Med Sci, 1995 Jun, 57(3), 499 - 502 Isolation and characterization of Staphylococcus aureus in rats trapped at restaurants in buildings in downtown Tokyo; Kato Y et al.; 165 (18.1%) out of 910 rats captured at restaurants in 14 buildings in downtown Tokyo were positive for Staphylococcus aureus . The 165 S . aureus strains isolated were biotyped into A, B, C, D, G, and untypable groups (UT1 and UT2) . The UT1 was the most frequent (72 strains, 43.6%), followed by biotype G (33 strains, 20.0%) . The strains were classified into coagulase types I, II, III, IV, V, VI, VII, VIII, and an untypable group . Coagulase type V was the most frequent (92 strains, 55.8%), followed by coagulase type VII (25 strains, 15.2%) . Enterotoxins A, B, C, or D were produced by 35 strains . Enterotoxins A and B were the most frequent (13 strains each) . Toxic shock syndrome toxin-1 was produced by 3 strains . 65 strains were resistant to ampicillin, 2 to oxytetracycline, and 1 to erythromycin. APMIS, 1995 Jun, 103(6), 460 - 8 Evaluation of a method for measurement of intracellular killing of Staphylococcus aureus in human neutrophil granulocytes; Nielsen SL et al.; A modified method for measurement of intracellular killing of Staphylococcus aureus in human neutrophil granulocytes is described . After phagocytosis of S . aureus the extracellular bacteria were sufficiently removed by repeated centrifugations and washings of the granulocytes . The optimal conditions for incubation of granulocytes for measurement of intracellular killing were found to be 37 degrees C in the presence of 5% CO2 . Under these conditions, stable pH, the viability and the capacity of the granulocytes for superoxide anion generation were preserved . The number of intracellular viable bacteria was determined after lysis of the granulocytes, which should be done in H2O at pH 11 to ensure sufficient cell lysis . The kinetics and individual variation of the intracellular killing are described . The intra- or extracellular location of surviving bacteria was studied . After approximately 8 h incubation we observed intracellular growth of S . aureus followed by lysis of granulocytes and extracellular growth of bacteria . Consequently, the incubation period should not be extended beyond 5 to 8 h when the assay is used to study the effects of antibiotics on intracellular killing. J Bacteriol, 1995 Jun, 177(11), 3220 - 6 Insertional inactivation of a chromosomal locus that modulates expression of potential virulence determinants in Staphylococcus aureus; Cheung AL et al.; A single insertion of transposon Tn551 into a unique chromosomal locus of Staphylococcus aureus ISP479C has resulted in a pleiotropic effect on the expression of both extracellular and cell wall proteins . In particular, the expression of cell wall protein A and clumping activity with fibrinogen were rendered undetectable in the mutant 1E3 compared with the parent . The secretion of alpha-hemolysin in mutant 1E3 was modestly increased . Southern blot and phenotypic analyses indicated that this locus is distinct from agr, xpr, and sar, three previously described global regulatory loci . Transduction experiments demonstrated that the genotype associated with mutant 1E3 could be transferred back into the parental strain ISP479C . The transductant 1E3-2 displayed a phenotypic profile similar to that of the original mutant . Northern (RNA) blot studies showed that this locus may be involved in modulating target genes at the mRNA level . In the rabbit endocarditis model, there was a significant decrease in both the infectivity rate and intravegetation bacterial density with mutant 1E3 compared with the parent at an inoculum of 10(3) CFU . Since protein A and the fibrinogen-binding protein(s) are major surface proteins that may mediate bacterial adhesion to host tissues, this locus may be an important genetic element involved in the expression of virulence determinants in S . aureus. Mol Microbiol, 1995 Jun, 16(5), 895 - 907 Identification of the ligand-binding domain of the surface-located fibrinogen receptor (clumping factor) of Staphylococcus aureus; McDevitt D et al.; The ability of Staphylococcus aureus to bind to fibrinogen and fibrin is believed to be an important factor in the initiation of foreign-body and wound infections . Recently, we reported the cloning and sequencing of the gene clfA encoding the fibrinogen receptor (clumping factor, ClfA) of S . aureus strain Newman and showed that the gene product was responsible for the clumping of bacteria in soluble fibrinogen and for the adherence of bacteria to solid-phase fibrinogen . This was confirmed here by showing that antibodies raised against purified Region A inhibited both of these properties . Also, immunofluorescent microscopic analysis of wild-type Newman and a clfA::Tn917 mutant of Newman with anti-ClfA Region A sera confirmed that Region A is exposed on the bacterial cell surface . Furthermore, polystyrene beads coated with the Region A protein formed clumps in soluble fibrinogen showing that the ClfA protein alone is sufficient for the clumping phenotype . Western immunoblotting with anti-ClfA Region A antibodies identified the native ClfA receptor as a 185 kDa protein that was released from the cell wall of S . aureus by lysostaphin treatment . A single extensive ligand-binding site was located within Region A of the ClfA protein . Truncated ClfA proteins were expressed in Escherichia coli . Lysates of E . coli and proteins that had been purified by affinity chromatography were tested for (i) their ability to bind fibrinogen in Western ligand blotting experiments, (ii) for their ability to inhibit clumping of bacteria in fibrinogen solution and adherence of bacteria to solid-phase fibrinogen, and (iii) for their ability to neutralize the blocking activity of anti-ClfA Region A antibody . These tests allowed the ligand-binding domain to be localized to a 218-residue segment (residues 332-550) within Region A. Mol Microbiol, 1995 Jun, 16(5), 877 - 93 Novel organization of the site-specific integration and excision recombination functions of the Staphylococcus aureus serotype F virulence-converting phages phi 13 and phi 42; Carroll D et al.; Functions required for site-specific integration and excision of the Staphylococcus aureus serotype F virulence-converting phages phi 13 and phi 42 were localized and characterized . Like other temperate phages, integration of phi 13 and phi 42 sequences was found to require the product of an int gene located close to the phage attP site . Both int genes are almost identical, express proteins possessing characteristic features of the Int (integrase) family of recombinases, but share very little homology with previously described int genes, including those of the serotype B S . aureus phages L54a and phi 11 . Nevertheless, all four S . aureus phages share an almost identical short sequence located immediately 5' to these distinct int genes, suggesting a common mechanism of int gene regulation . Upstream from these common sequences, the sequences of phi 13 and phi 42 are quite distinct from each other, and from the corresponding regions of phi 11 and L54a which encode the Xis proteins that are required with Int to mediate site-specific excision of the latter phages . Surprisingly, phi 13 and phi 42 sequences encompassing the attP sites and int genes, but lacking either an adjacent or more distant phage excision protein gene, were sufficient to mediate site-specific excision of integrated phage DNA sequences. FEBS Lett, 1995 May 29, 365(2-3), 193 - 7 The tetracycline efflux protein encoded by the tet(K) gene from Staphylococcus aureus is a metal-tetracycline/H+ antiporter; Yamaguchi A et al.; The tet(K) gene from Staphylococcus aureus was highly expressed in Escherichia coli by an alteration of its initiation codon from TTG to ATG and its ribosome-binding sequence from GAGG to GGAGG {Noguchi, N . et al . (1994) Biol . Pharm . Bull . 17, 352-355} . The inverted membrane vesicles prepared from the tet(K)-expressing cells showed respiration-dependent {3H}tetracycline transport comparable to the vesicles from the tet(B)-expressing cells . The affinity of Tet(K) vesicles to tetracycline was the same as that of Tet(B) vesicles, whereas the former Vmax value was about 60% of the latter one . Contrary to Tet(B) vesicles, Tet(K) vesicles showed no significant minocycline uptake, which was consistent with the low minocycline resistance of the Tet(K)-producing cells . The tetracycline transport mediated by Tet(K) vesicles was coupled with proton transport and the translocation of 60Co2+ ions as well as in Tet(B) vesicles . This observation indicates that the class K tetracycline resistance determinant from Gram-positive bacteria also encodes a metal-tetracycline/H+ antiporter that is functionally similar to that encoded by tet(B), although there is a considerable difference in the primary sequences and the putative topologies of these Tet proteins. J Biol Chem, 1995 May 26, 270(21), 12839 - 45 The Staphylococcus aureus enterotoxin B superantigen induces specific T cell receptor down-regulation by increasing its internalization; Niedergang F et al.; Superantigens are able to stimulate T lymphocyte populations expressing T cell antigen receptors (TCR) belonging to particular V beta families . Moreover, the presence of these superantigens may induce long term unresponsiveness (anergy) of these sensitive cells . Some bacterial toxins are potent superantigens . We have analyzed in vitro the capacity of some Staphylococcus aureus enterotoxin superantigens to modulate T cell antigen receptor expression and the cellular mechanisms involved . Staphylococcus enterotoxin B (SEB) induced rapid down-regulation of surface T cell antigen receptors in V beta 3-expressing T lymphocytes, as assessed by flow cytometry . This phenomenon was a consequence of the direct interaction between the toxin and the TCR since it was observed in the absence of cells expressing major histocompatibility complex class II molecules . The cellular mechanism involved in SEB-induced down-regulation of TCR was further investigated . Immunofluorescence and confocal microscopy experiments showed that toxin B induced intracellular accumulation of TCR.CD3 in endocytic vesicles . Moreover, SEB induced an increase in T cell receptor endocytosis as measured using radiolabeled Fab fragments of an anti-CD3 monoclonal antibody . Taken together, our observations indicate that Staphylococcus enterotoxin B superantigen induced changes in the dynamics of surface T cell receptors, which resulted in the fast reduction of membrane receptor numbers. J Biol Chem, 1995 May 19, 270(20), 12005 - 11 Critical residues in the ligand-binding site of the Staphylococcus aureus collagen-binding adhesin (MSCRAMM); Patti JM et al.; We have identified a discrete collagen-binding site within the Staphylococcus aureus collagen adhesin that is located in a region between amino acids Asp209 and Tyr233 . Polyclonal antibodies raised against a recombinant form of the collagen adhesin inhibited the binding of collagen type II to S . aureus . When overlapping synthetic peptides mimicking segments of the adhesin fragment were tested for their ability to neutralize the inhibitory activity of the antibody only one peptide, CBD4 was found to be active . CBD4 bound directly to collagen and at high concentrations inhibited the binding of collagen to S . aureus . A synthetic peptide derivative of CBD4 lacking 2 carboxyl-terminal residues (Asn232, Tyr233) had no inhibitory activity . The importance of these residues for collagen binding was confirmed by biospecific interaction analysis . Mutant adhesin proteins N232-->A and Y233-->A exhibited dramatic changes in collagen binding activity . The dominant dissociation rate for the binding of mutant adhesin protein N232-->A to immobilized collagen II decreased almost 10-fold, while the Y233-->A and the double mutant exhibited even more significant decreases in affinity and apparent binding ratio when compared to the wild type protein. J Biol Chem, 1995 May 12, 270(19), 11348 - 57 Photolabeling of a pore-forming toxin with the hydrophobic probe 2-{3H}diazofluorene . Identification of membrane-inserted segments of Staphylococcus aureus alpha-toxin; Lala AK et al.; The identification of membrane-inserted segments of pore-forming soluble proteins is crucial to understanding the action of these proteins at the molecular level . A distinct member of this class of proteins is alpha-toxin, a 293-amino acid-long 33-kDa hemolytic toxin secreted by Staphylococcus aureus that can form pores in both artificial and natural membranes . We have studied the interaction of alpha-toxin with single bilayer vesicles prepared from asolectin using a hydrophobic photoactivable reagent, 2-{3H}diazofluorene ({3H}DAF) (Pradhan, D., and Lala, A . K . (1987) J . Biol . Chem . 262, 8242-8251) . This reagent readily partitions into the membrane hydrophobic core and on photolysis labels the lipid and protein segments that penetrate the membrane . Current models on the mode of action of alpha-toxin indicate that, on interaction with membranes, alpha-toxin forms an oligomer, which represents the active pore . In keeping with these models, we observe that {3H}DAF photolabels the membrane-bound alpha-toxin oligomer . Cyanogen bromide fragmentation of {3H}DAF-labeled alpha-toxin gave several fragments, which were subjected to Edman degradation . We could thus sequence residues 1-19, 35-60, 114-139, 198-231, and 235-258 . Radioactive analysis and phenylthiohydantoin-derivative analysis during sequencing permitted analysis of DAF insertion sites . The results obtained indicated that the N and C termini (residues 235-258) have been extensively labeled . The putative pore-forming glycine-rich central hinge region was poorly labeled, indicating that the apposing side of the lumen of the pore does not form the lipid-protein interface . The DAF labeling pattern indicated that the major structural motif in membrane-bound alpha-toxin was largely beta-sheet. J Biochem (Tokyo), 1995 May, 117(5), 974 - 9 Binding of myosin and its subfragment-1 with antibodies specific to the two heads of the myosin molecule; Murai S et al.; It was shown by Miyanishi et al . {Miyanishi, T., Maita, T., Matsuda, G., and Tonomura, Y . (1982) J . Biochem . 91, 1845-1853} that the amino acid sequence around the reactive lysine residue is different between head B (Pi-burst head) and head A of the myosin molecule . Thus, we synthesized these two peptides, and prepared rabbit polyclonal antibodies against them . Each antibody bound strongly with both peptides . However, the binding of the antibodies with S-1 was inhibited by the peptide used for the antigen but unaffected by the non-antigen peptide, suggesting that only antibodies specific to each head can bind with S-1 . Myosin was absorbed by either antibody A or B, which was immobilized on protein A in Staphylococcus aureus cells . However, half of S-1 was absorbed by each of the antibodies . The S-1 prepared showed about 0.5 mol of initial Pi-liberation per mol of S-1 . The Pi-burst size of S-1 unbound to the immobilized anti-A antibody increased to almost 1 mol/mol S-1, while that of S-1 unbound to the anti-B antibody decreased to 0.15 mol/mol S-1 . These results suggest the existence of two kinds of heads in the myosin molecule. Zentralbl Veterinarmed B, 1995 May, 42(3), 129 - 39 Characteristics of Staphylococcus aureus strains isolated from bovine mastitic milk: bacteriophage and antimicrobial agent susceptibility, and enterotoxigenicity; Adesiyun AA; Staphylococcus aureus strains isolated from bovine mastitic milk in Trinidad were examined for their susceptibility to bacteriophages and antimicrobial agents and their ability to produce enterotoxins . Phage 42D was used to screen for bovine strains of S . aureus in milk . Of 250 strains tested, 224 (89.6%) were sensitive to phages in the international phage set (IPS), 85 (34.0%) were resistant to antimicrobial agents and 134 (53.6%) were enterotoxigenic . Strains lysed by phages in various groups (mixed) were prevalent, 145 (58.0%), followed by strains sensitive to groups III (17.0%) and I (8.8%) phages . A total of 72 (28.8%) strains were lysed by phage 42D either alone or with others . Resistance to penicillin was most common with 59 (23.6%) strains while 44 (17.6%) and 43 (17.2%) strains were resistant to ampicillin and triple sulphur respectively . Only 3 (1.2%) strains were resistant to methicillin . Prevalence of resistance to penicillin (12.5%) amongst phage 42D-sensitive strains was significantly (P < or = 0.01; X2) lower than for strains not lysed by phage 42D (28.1%) but strains susceptible to phage 42D were significantly (P < or = 0.05; X2) more resistant (4.2%) to methicillin than those not lysed by the phage (0.0%) . Amongst 134 enterotoxigenic strains, 32 (23.9%), 77 (57.5%), 67 (50.0%) and 21 (15.7%) produced staphylococcal enterotoxins A(SEA), B(SEB), C(SEC) and D(SED) respectively either alone or mixed . SEB and SEC were significantly (P < or = 0.01; X2) more produced than either SEA or SED . Strains lysed by groups IV, i.e . 42D (62.5%), and III (56.7%) were more enterotoxigenic than those sensitive to phages in groups II (45.5%) and non-typable (46.2%) but the differences were not statistically significant (P > or = 0.05; X2) . Strains lysed by group II phages (72.7%) were significantly (P < or = 0.05; X2) more resistant to antimicrobial agents than those lysed by phage 42D (18.8%) . It was concluded that bovine mastitis strains of S . aureus in Trinidad were highly susceptible to bacteriophages and antimicrobial agents and enterotoxigenic and less than one-third may be considered to be bovine strains. Med Microbiol Immunol (Berl), 1995 May, 184(1), 33 - 6 Hemagglutination by Staphylococcus aureus strains responsible for human bacteremia or bovine mastitis; Rupp ME et al.; Although hemagglutination by Staphylococcus aureus has been associated with the pathogenesis of bovine mastitis, this trait has not been characterized with regard to human disease . In this study, the prevalence of hemagglutination in 100 strains of S . aureus responsible for bovine mastitis or human bacteremia, was characterized . Under optimum conditions hemagglutination was noted in 23% of the bovine strains, but only 13% of human strains, leading us to conclude that this trait is not a significant virulence determinant in human systemic infection . Additional studies indicate the hemagglutinin of S . aureus strains responsible for human bacteremia is proteinaceous in character. Rinsho Byori, 1995 May, 43(5), 487 - 92 {Detection of heterogeneous MRSA by using the PCR method and population analysis}; Moriki S et al.; To assess the clinical usefulness of the PCR method for the detection of methicillin-resistant Staphylococcus aureus (MRSA) by targetting the mecA gene, we surveyed 150 clinical isolates of Staphylococcus aureus and compared the results of the PCR method with those of the standard broth microdilution method . Fifty-four isolates (36%) were positive for the mecA gene and two of them, presenting coagulase type IV, were recognized as susceptible strains by microdilution method, while all the other mecA positive strains were drug-resistant . Population analysis revealed that these two strains were heterogeneous in terms of drug-resistance and composed of two populations of cells; i.e., relatively susceptible cells and highly resistant cells . The discrepancy between the phenotypic expression (drug-resistance) and the genotype (mecA gene) seems to be due to the small percentage of highly resistant cells . Drug-resistant colonies could be induced in these strains by the contact with methicillin, indicating the selective increase of the population of resistant cells by a passage in the drug . These observations suggest that it is clinically important to detect the resistance-inducible strains (prototype MRSA) by using the PCR method and population analysis. J Cardiol, 1995 May, 25(5), 263 - 8 {Right-sided infectious endocarditis due to methicillin-resistant Staphylococcus aureus resulting in ruptured abscess of the ventricular septum and sinus of Valsalva}; Nishinaga M et al.; A 24-year-old woman presented with right-sided infectious endocarditis due to methicillin-resistant Staphylococcus aureus (MRSA) . This is the first report of right-sided infectious endocarditis caused by MRSA in Japan . The patient was admitted to the Jichi Medical School Hospital because of fever of unknown origin and disturbance of consciousness . Several months before, she had discontinued treatment for hyperthyroidism . Antibiotics effective against MRSA, vancomycin and flomoxef, were given intravenously, but a new heart murmur was detected . Echocardiographic study revealed vegetations attached to the tricuspid valve and abscess formation on the ventricular septum . The vancomycin dosage was increased and arbekacin sulfate was also given from the sixth hospital day . However, these antibiotics had very little effect and the abscess rapidly increased in size . Color flow mapping finally demonstrated intracardiac shunt flow through the ruptured abscess of the ventricular septum and sinus of Valsalva . She died suddenly, probably from heart failure . The prognosis of infectious endocarditis due to MRSA is poor and drug therapy often fails . Thus, surgery should be considered in the early stage. Microbiology, 1995 May, 141 ( Pt 5), 1255 - 65 Typing of Staphylococcus aureus strains by PCR-amplification of variable-length 16S-23S rDNA spacer regions: characterization of spacer sequences; Gurtler V et al.; To develop a rapid and accurate method of typing large numbers of clinical isolates of Staphylococcus aureus, the spacer region C of the rRNA operon {1391-507 (16S-23S)} was enzymically amplified from 322 strains . When the products were separated by denaturing PAGE, 15 variable-length rrn alleles were demonstrated, ranging in size from 906 to 1223 bp . The variable-length HpaII-digested region C {(region E; 1446-196 (16S-23S)} amplification products were cloned into M13mp18RF to sequence separate variable-length alleles . A total of 17 region E inserts were sequenced, aligned and divided into nine alleles by length (938-1174) and sequence properties . The 16S-23S spacer rDNA varied in length (303-551 bp) and in properties; three alleles contained a tRNAIle gene alone, two alleles contained a tRNAIle and a tRNAAla gene, and four alleles lacked tRNA genes . The sequences of two alleles showed less than 1% variation when isolated from two or three S . aureus strains . The 48 penicillin- and methicillin-sensitive strains were divided into 26 ribotypes; in contrast, the 274 methicillin-resistant S . aureus (MRSA) strains were divided into nine ribotypes (A-I) with 97% typing as either ribotype A or B (rrnL was missing in B) . The sequence conservation of the rrn operons argues for the use of the 16S-23S spacer region as a stable and direct indicator of the evolutionary divergence of S . aureus strains. Curr Microbiol, 1995 May, 30(5), 299 - 303 Salt-induced cell lysis of Staphylococcus aureus; Yabu K et al.; Cell lysis was efficiently induced in Staphylococcus aureus by the addition of 0.3 M NaCl to exponentially growing cultures at 30 degrees C . When cells harvested at the exponential phase were incubated in buffer with NaCl, autolysis occurred . Treatment with chloramphenicol failed to induce cell lysis by NaCl . The effects of NaCl on growing cells and harvested cells were inhibited by the addition of sodium polyanethole sulfonate, subtilisin, cardiolipin, and lipoteichoic acid . These agents diminished the activity of a cell wall-lytic enzyme liberated from the cells in the presence of NaCl . Lysis induced by salt appears to be catalyzed by a similar lytic enzyme in growing and harvested cells. J Trauma, 1995 May, 38(5), 794 - 801 Endotoxemia and specific antibody behavior against different endotoxins following multiple injuries; Hiki N et al.; The aim of this study was to establish the incidence of endotoxemia and the influence of endotoxin on specific antibody response after multiple injury . Blood samples were collected from 39 patients (median Injury Severity Score: 20.5) at 0-3 and 6-12 hours, and 1, 3, 5, and 10 days after admission . The endotoxin plasma levels were high at the first time point (mean = 0.421 endotoxin units/mL) and decreased in the later course . Total immunoglobulin levels of IgM, IgG, or IgA were low and increased throughout the observation period . Specific antibodies of the IgM class against two lipid A and four lipopolysaccharide preparations increased transiently but significantly on day 3 and/or day 5 . No changes of specific antibody content against endotoxin or lipid A was seen in the IgG or IgA class . The specific antibody content of the different classes against alpha-hemolysin of Staphylococcus aureus did not differ during 10 days after trauma . The specific antibodies of the IgM class reacted with all lipid A and LPS lipopolysaccharide preparations demonstrating cross-reactivity . These results suggest that endotoxin may be a specific stimulator of IgM antiendotoxin antibody secretion following trauma. J Infect Dis, 1995 May, 171(5), 1230 - 6 Development and characterization of a new model of hematogenous osteomyelitis in the rat; Hienz SA et al.; Hematogenous osteomyelitis was produced in the tibia or the mandible of rats by drilling a hole into the bone, injecting sodium morrhuate, and inoculating Staphylococcus aureus Phillips into the femoral vein . Animals were sacrificed after 2 weeks and examined . The infection was characterized grossly and radiographically by bone deformation, histopathologically by a characteristic suppurative reaction, and microbiologically by the recovery of S . aureus Phillips from the infected tissue . These findings indicate that the model mimics human osteomyelitis with respect to its inflammatory bone changes . In contrast to earlier rat models in which bacteria were injected directly into the bone, this new experimental model allows study of the initiating events of osteomyelitis such as bacterial attachment and might assist as a model for both prophylactic and therapeutic trials. J Bacteriol, 1995 May, 177(10), 2609 - 14 In vitro transcription of pathogenesis-related genes by purified RNA polymerase from Staphylococcus aureus; Rao L et al.; The RNA polymerase (RNAP) holoenzyme of Staphylococcus aureus was purified by DNA affinity, gel filtration, and ion-exchange chromatography . This RNAP contained four major subunits with apparent molecular masses of 165, 130, 60, and 47 kDa . All four subunits of the RNAP were serologically related to the subunits of Escherichia coli E sigma 70 holoenzyme by Western immunoblot analysis . The 60-kDa subunit was subsequently isolated and found to react with a monoclonal antibody specific to the E . coli sigma 70 subunit . This sigma 70-related protein allowed E . coli core RNAP promoter-specific initiation and increased transcription by S . aureus RNAP that is unsaturated with sigma . We therefore suggest that this 60-kDa protein is a sigma factor . Purified S . aureus RNAP transcribed from the promoters of several important S . aureus virulence genes (sea, sec, hla, and agr P2) in vitro . The in vitro transcription start sites of the sea, sec, and agr P2 promoters, mapped by primer extension, were similar to those identified in vivo . The putative promoter hexamers of these three genes showed strong sequence similarity to the E . coli sigma 70 consensus promoter, and transcription by E sigma 70 from some of these promoters has been observed . Conversely, S . aureus RNAP does not transcribe from all E . coli sigma 70-dependent promoters . Taken together, our results indicate that the promoter sequences recognized by purified S . aureus RNAP are similar but not identical to those recognized by E . coli E sigma 70. South Med J, 1995 May, 88(5), 586 - 90 Pneumonia due to Staphylococcus aureus in a patient with AIDS: review of incidence and report of an atypical roentgenographic presentation; Dicpinigaitis PV et al.; Multiple defects in host defense mechanisms produce an increased incidence of community-acquired bacterial pneumonia in individuals infected with the human immunodeficiency virus . Clinical studies suggest that Staphylococcus aureus is an uncommon cause of such infections, though its incidence is increased in the setting of intravenous drug use, indwelling vascular catheter, and coexistent pulmonary Kaposi's sarcoma or pneumonia due to Pneumocystis carinii . The significantly higher incidence of S aureus pneumonia documented in autopsy series suggests that the infection frequently remains undiagnosed ante mortem . The clinical and radiologic presentation of staphylococcal pneumonia in HIV-seropositive patients is similar to that seen in immunocompetent hosts . However, atypical radiographic patterns can occur . We describe a case of S aureus pneumonia manifested as an infiltrate with focal predominance and multiple cavitary lesions . Such a radiologic appearance has not previously been described in this population . Given the likelihood that pneumonia due to S aureus is significantly underdiagnosed ante mortem, a high index of clinical suspicion is warranted. Infect Immun, 1995 May, 63(5), 1914 - 20 Cloning, expression, and nucleotide sequence of a Staphylococcus aureus gene (fbpA) encoding a fibrinogen-binding protein; Cheung AI et al.; Septicemia due to Staphylococcus aureus often begins as a focal infection (e.g., colonized wounds or catheters) from which the organism gains access to the bloodstream . On the basis of recent data from this laboratory, it is likely that S . aureus colonizes catheters and endothelium by using a fibrinogen-binding protein to mediate adhesion to fibrinogen-coated surfaces . To characterize the fibrinogen-reactive protein, we screened a lambda Zap library of S . aureus DB, a clinical isolate, for clones that were reactive with fibrinogen . Of 100,000 plaques screened, 3 were found to react with fibrinogen on immunoblots . Plasmid DNA prepared from clones 14, 30, and 36, upon digestion with EcoR1, which released the insert, revealed fragments of 4.6, 3.6, and 3.2 kb, respectively . To identify the cloned protein expressed in E . coli, cells were fractionated into periplasmic, membrane, and cytoplasmic fractions . Expression studies of clone 14, which comprised approximately two-thirds of the mature molecule, including the C terminus, revealed a 34-kDa fibrinogen-reactive protein in both the periplasmic and membrane fractions . This protein, designated FbpA, could be partially purified on a fibrinogen column . By using both clones 14 and 36 as templates, the complete DNA sequence of the fibrinogen-binding protein was obtained, yielding a molecule with a predicted size of 69,991 Da . Although sequence analysis revealed a high degree of homology with coagulase, there is a unique sequence of 11 amino acids that is not found in three known coagulases as well as two recently cloned fibrinogen-binding proteins . This unique sequence shares homology with a cell wall anchor motif found in other gram-positive surface proteins. Infect Immun, 1995 May, 63(5), 1835 - 9 Internalization of Staphylococcus aureus by endothelial cells induces cytokine gene expression; Yao L et al.; The ability of the vascular endothelium to elaborate cytokines in response to gram-positive sepsis has received limited attention . This study examined cytokine expression by human umbilical vein endothelial cells (EC) following infection with a gram-positive bacterial pathogen, Staphylococcus aureus . S . aureus infection of EC resulted in the production of interleukin-6 (IL-6) and IL-1 beta . For IL-6, message was detected at 3 h after infection, protein was present at 24 h, and both message and protein persisted for 72 h . IL-1 beta message was detected at 12 h, IL-1 beta protein was detected at 24 h, and both persisted for 72 h . Message for colony-stimulating factor 1 remained unaltered . UV-killed S . aureus also elicited IL-1 beta and IL-6 message and protein expression at 24 and 48 h . Twenty-one clinical isolates of S . aureus were tested, and all induced IL-6 release by 48 h . However, the laboratory strain 8325-4 did not induce cytokine expression at any time point and was internalized by EC 1,000-fold less than other strains were . Internalization of latex beads by EC did not induce IL-6 gene expression . Furthermore, cytochalasin D treatment of the EC prevented IL-1 and IL-6 induction by S . aureus but not by tumor necrosis factor alpha or lipopolysaccharide . These results indicate that S . aureus is a potent inducer of IL-1 and IL-6 in EC and that internalization of S . aureus by EC is necessary for their cytokine expression . Thus, our data suggest that the vascular endothelium may play an important role in the pathogenesis of septicemia caused by gram-positive organisms. Rev Med Chil, 1995 May, 122(5), 487 - 95 {Molecular study of 6 episodes of nosocomial infections produced by methicillin resistant Staphylococcus aureus}; Kaltwasser G et al.; A critical step in any epidemiologic research concerning nosocomial infections is the precise identification of the responsible pathogen . The present work utilized a molecular approach -plasmids identification, restriction length polymorphism DNA analysis, and random amplified polymorphic DNA- for the characterization of 6 nosocomial outbreaks due to 52 strains of methicillin-resistant Staphylococcus aureus (MRSA) . In these episodes, the clinic-epidemiologic and phenotypic analysis (antibiotype) pointed to a nosocomial infection . Through molecular analysis it was possible to establish, in a very precise way, clonality due to MRSA strains in 2 of the studied outbreaks; the same type of analysis allowed to eliminate a MRSA clonal origin in the remainder 4 episodes . The antibiogram was not an useful analytic tool due to its poor discriminatory power . Also, through a PCR procedure, it was possible to identify the presence of the gen mecA in every of the 52 MRSA strains studied. Obstet Gynecol, 1995 May, 85(5 Pt 2), 834 - 5 Sacroiliitis associated with pyelonephritis in pregnancy; Egerman RS et al.; BACKGROUND: Sacroiliitis is a rare infection and an unusual cause of back pain during pregnancy . Because pregnancy and infections commonly associated with pregnancy are risk factors, this diagnosis should be considered in the gravida with sacroiliac pain . CASE: A 17-year-old woman at 24 weeks' gestation, with a history of illicit drug use, presented to a local emergency room with back and buttock pain . Bacteriuria and pyuria were diagnosed, and cefazolin was initiated . Blood cultures grew Staphylococcus aureus and Escherichia coli . Despite prolonged antibiotic therapy for possible endocarditis, she had persistent debilitating lower back and buttock pain . Radiographic studies detected sacroiliitis, and broadened antibiotic therapy effected cure . CONCLUSION: When sacroiliitis is suspected, diagnostic imaging with either computed axial tomography, or, preferably, magnetic resonance imaging may be helpful . Antibiotic therapy should be tailored to the specific organism involved and continued for 3-6 weeks. J Immunol, 1995 May 1, 154(9), 4341 - 50 The role of IL-10 in human B cell activation, proliferation, and differentiation; Itoh K et al.; Recent studies have disclosed variable effects of IL-10 on viabilities of human B lineage cells . Thus, IL-10 has been shown to prevent apoptosis of germinal center B cells, whereas IL-10 has been found to induce apoptosis of B-chronic lymphocytic leukemia cells, suggesting the possibility that the effects of IL-10 might be different depending on the state of activation of B cells . The current studies therefore examined in detail the regulation of the survival of human peripheral blood B cells by IL-10 and its relevance to Ig production . Highly purified B cells from healthy adult individuals were cultured with Staphylococcus aureus (SA) Cowan I in the presence or absence of IL-10 . When IL-10 was present during the initial activation of B cells with SA, IL-10 facilitated the apoptosis of SA-activated B cells, as determined by staining with propidium iodide, followed by analysis with flow cytometry, thus resulting in very modest IgM production . IL-2 prevented the IL-10-mediated progression of the apoptosis of SA-activated B cells during the initial activation, and thus restored the further differentiation of these B cells into Ig secreting cells . By contrast, IL-10 rather rescued SA-activated B cells from apoptosis and thus supported the differentiation of these B cells without any influences of IL-2, when it was added after 72 h of culture . Of note, cyclosporin A prevented the IL-10-mediated promotion of the apoptosis of SA-activated B cells, thus resulting in the marked enhancement of IgM production of B cells stimulated with SA + IL-10 . Finally, the promotion or prevention of the IL-10-mediated apoptosis was correlated with the expression of Bcl-2 oncoprotein in SA-activated B cells . These results indicate that the effects of IL-10 are different depending on the state of activation of B cells after ligation of Ag receptors . Thus, the data have demonstrated that IL-10 during the initial activation delivers negative signals that promote the apoptosis of B cells, whereas IL-10 supports the differentiation of B cells in the complete absence of IL-2 during the subsequent responses following activation . These results therefore emphasize unique biphasic effects of IL-10 on human B cell responsiveness in determining the outcome of humoral immune responses. J Infect, 1995 May, 30(3), 227 - 33 Effect of glucose in dialysis fluid on antibacterial defence in the peritoneal cavity; Calame W et al.; To study the effect of glucose concentration and dwell time of dialysis fluid on peritoneal antibacterial defence, an experimental infection with Staphylococcus aureus was induced in rats . For this purpose rats were inoculated intraperitoneally with Staphylococcus aureus at different intervals after the administration of various dialysis fluids . Twenty-four hours later the numbers of bacteria and cells in the peritoneal cavity were determined . The number of bacteria was correlated positively with the glucose concentration . Furthermore, an inverse correlation between dwell time and the number of bacteria was observed . Neither finding could be attributed to a glucose-dependent growth of the bacteria or disruption of the killing capacity of peritoneal cells in vitro . A glucose-dependent increase in the volume of the peritoneal fluid could partially explain the differences found in vivo . It is concluded that the glucose in dialysis fluid impairs antibacterial defence in the peritoneal cavity and that longer dwell times enhance this defence. J Hosp Infect, 1995 May, 30(1), 39 - 49 Nasal carriage of Staphylococcus aureus and cross-contamination in a surgical intensive care unit: efficacy of mupirocin ointment; Talon D et al.; A six month prospective study was carried out in a surgical intensive care unit (SICU) of a university hospital to assess the incidence and routes of exogenous colonization by Staphylococcus aureus . A total of 157 patients were included in the study . One thousand one hundred and eleven specimens (nasal, surgical wound swabs, tracheal secretions obtained on admission and once a week thereafter, and all clinical specimens) were collected over a four month period from patients without nasal decontamination (A) . They were compared with 729 specimens collected over a two month period from patients treated with nasal mupirocin ointment (B) . All S . aureus strains were typed by restriction fragment length polymorphism (RFLP) pulsed-field gel electrophoresis after SmaI macrorestriction . The nasal colonization rates on admission were 25.5 and 32.7% in groups A and B, respectively . Thirty-one untreated patients (31.3%) and three patients (5.1%) treated with nasal ointment, acquired the nasal S . aureus in the SICU (P = 0.00027) . Nasal carriers were more frequently colonized in the bronchopulmonary tract (Bp) and surgical wound (Sw) (62%) than patients who were not nasal carriers (14%) (P < 0.00001) . The patterns were identical for nasal, Bp and Sw strains from the same patient . RFLP analysis characterized seven epidemic strains of methicillin-resistant S . aureus (MRSA) which colonized 60% of group A and 9% of group B patients (P < 0.00001) . The bronchopulmonary tract infection rate was reduced in group B (P = 0.032) . In conclusion, in an SICU, nasal carriage of S . aureus appeared to be the source of endogenous and cross-colonization . The use of nasal mupirocin ointment reduced the incidence of Bp and Sw colonization, as well as the MRSA infection rate. Rinsho Shinkeigaku, 1995 May, 35(5), 537 - 41 {Acute bacterial meningitis and foramen magnum syndrome as a presentation of odontoid osteomyelitis . A case report}; Akanuma J et al.; A 77-year-old male was hospitalized with a 6-day history of severe posterior cervical pain and headache . An initial lumbar puncture revealed polymorphonuclear pleocytosis indicating pyogenic meningitis . The blood and throat cultures on admission were positive for Staphylococcus aureus . A combination of ceftriaxone and ABPC, both of which were sensitive to the pathogen, were intravenously administered for one month, by which he responded well as to the meningitis . However, he was found to have persistent neck pain, and wasting and weakness of the shoulder girdles, which were associated with polyneuropathy-like sensory loss . Neuroimaging studies disclosed a partial destruction of the odontoid process, subluxation of the atlantoaxial joint, MRI evidence of an inflammatory pseudomass formation dorsal to the odontoid process and an increased uptake of radionuclide on bone scanning, all of which were indicative of osteomyelitic processes at the region of the axis; i.e., the meningitis was thought to be secondary to osteomyelitis of the odontoid process and hence the foramen magnum syndrome characteristic to this particular case was explained by the compression of the cervicomedullary junction due to the pseudomass . To our knowledge, this is the first case of primary odontoid osteomyelitis to be reported in Japan. Protein Sci, 1995 May, 4(5), 936 - 44 Investigation of a side-chain-side-chain hydrogen bond by mutagenesis, thermodynamics, and NMR spectroscopy; Hammen PK et al.; Anomalous NMR behavior of the hydroxyl proton resonance for Ser 31 has been reported for histidine-containing protein (HPr) from two microorganisms: Escherichia coli and Staphylococcus aureus . The unusual slow exchange and chemical shift exhibited by the resonance led to the proposal that the hydroxyl group is involved in a strong hydrogen bond . To test this hypothesis and to characterize the importance of such an interaction, a mutant in which Ser 31 is replaced by an alanine was generated in HPr from Escherichia coli . The activity, stability, and structure of the mutant HPr were assessed using a reconstituted assay system, analysis of solvent denaturation curves, and NMR, respectively . Substitution of Ser 31 yields a fully functional protein that is only slightly less stable (delta delta G(folding) = 0.46 +/- 0.15 kcal mol-1) than the wild type . The NMR results confirm the identity of the hydrogen bond acceptor as Asp 69 and reveal that it exists as the gauche- conformer in wild-type HPr in solution but exhibits conformational averaging in the mutant protein . The side chain of Asp 69 interacts with two main-chain amide proteins in addition to its interaction with the side chain of Ser 31 in the wild-type protein . These results indicate that removal of the serine has led to the loss of all three hydrogen bond interactions involving Asp 69, suggesting a cooperative network of interactions . A complete analysis of the thermodynamics was performed in which differences in side-chain hydrophobicity and conformational entropy between the two proteins are accounted for.(ABSTRACT TRUNCATED AT 250 WORDS) Zh Mikrobiol Epidemiol Immunobiol, 1995 May-Jun, (3), 109 - 13 {The immunomorphological characteristics of experimental bacterial sepsis}; Morozov VL et al.; The experimental model of bacterial sepsis, based on the intraperitoneal inoculation of Staphylococcus aureus with the simultaneous suppression of sensitivity by the injection of prednisolone, has been developed in 40 guinea pigs . Sepsis is characterized by the generalization of the infection in the body and by the formation of disseminated purulent foci simultaneously with inflammatory and dystrophic changes in different organs . The development of experimental sepsis leads to an increase in the content of B lymphocytes and to the activation of humoral immunity simultaneously with the suppression of cell-mediated immunity. J Pharm Sci, 1995 May, 84(5), 661 - 4 Synthesis and antimicrobial activity of 6,7-annulated pyrido{2,3-d}pyrimidines; Donkor IO et al.; Four new 6H-indeno{2',1':5,6}pyrido{2,3-d}pryimidines (10-13) were synthesized via cyclocondensation reactions involving chlorovinyl aldehyde 1 or ketoaldehyde 3 and appropriately substituted 6-aminopyrimidines . The regiochemistry of the compounds was established by 1H NMR and 13C NMR spectral data as well as X-ray crystal data . Compounds 10 and 11 and previously reported homologues 14 and 15 were screened for antimicrobial activity . Moderate antimicrobial activity was observed for some of these compounds . Compound 14 was especially active against Staphylococcus aureus . Crystal data for 13 (C14H7N3Cl2) follows: monoclinic space group, P21/n; Unit cell dimensions, a = 7.284(1) A, b = 12.800(1) A, c = 13.108(1) A, beta = 93.98(1) degree, V = 1219.2(2) A3, Z = 4. Infect Control Hosp Epidemiol, 1995 May, 16(5), 260 - 7 Molecular epidemiology of methicillin-resistant Staphylococcus aureus at a low-incidence hospital over a 4-year period; Lugeon C et al.; OBJECTIVE: To study the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) over a prolonged period of time with the aid of a molecular typing method (ribotyping) . SETTING: A 1,000-bed tertiary university medical center . PATIENTS AND METHODS: Defined epidemiological data were recorded for all patients culture-positive for MRSA between 1989 and 1992 . Ribotyping of MRSA strains was performed using three restriction enzymes: EcoRV, HindIII, and KpnI . RESULTS: From 1989 to 1992, MRSA was isolated from clinical specimens in 98 patients and from surveillance cultures in 27 patients . Among the 122 isolates available for typing, 26 different ribotypes were identified . In 20% of the cases, MRSA was community-acquired, and a third of these patients never had been hospitalized previously . Nine ribotypes were responsible for more than one case (2 to 64 patients); 17 appeared only once . Epidemiological data correlated with ribotyping results revealed 14 epidemiologic clusters involving six different ribotypes, whereas only three outbreaks were suspected initially . The median follow-up after the last isolation of a given ribotype was 14 months (range, 1 to 42) for clusters and 25 months (range, 1 to 46) for ribotypes that appeared only once . During clusters, only 16% of the cases occurred after the implementation of control measures in the ward (breakthrough cases) . CONCLUSIONS: The high diversity of MRSA strains observed over 4 years suggested that new strains were introduced continuously in our hospital . Furthermore, that 17 ribotypes were isolated only once, that breakthrough cases represented only 16% of the cases in clusters, and that the follow-up duration after the last isolation of a given ribotype was more than 14 months suggest that infection control measures were effective in limiting the nosocomial spread of MRSA over a prolonged period of time. Chemotherapy, 1995 May-Jun, 41(3), 172 - 7 Degradation of penicillin-binding protein 2' in methicillin-resistant Staphylococcus aureus; Sumita Y et al.; We detected a novel protein with {14C}benzylpenicillin (PCG)-binding capacity and a molecular mass of about 60 kD in methicillin-resistant Staphylococcus aureus using a high concentration of {14C}PCG and extending the reaction time . However, the fluorogram showed that the band density of penicillin-binding protein 2' (PBP2') decreased gradually with incubation time . The appearance of the 60-kD protein and the reduction of the band density of PBP2' were stoichiometrically linked, and the binding profiles of beta-lactams for PBP2' and the 60-kD protein corresponded . These results suggested that the 60-kD protein is a degradation product of PBP2'. Br J Pharmacol, 1995 May, 115(1), 147 - 57 Effects of GTP gamma S on muscarinic receptor-stimulated inositol phospholipid hydrolysis in permeabilized smooth muscle from the small intestine; Prestwich SA et al.; 1 . Smooth muscle fragments from the longitudinal layer of the small intestine of the guinea-pig were permeabilized with Staphylococcus aureus alpha toxin (alpha-toxin) and used to investigate the role of G-protein activation in the regulation of muscarinic acetylcholine receptor (AChR)-stimulated inositol phospholipid hydrolysis . 2 . The efficiency of alpha-toxin permeabilization was estimated by the release of {3H}-2-deoxyglucose ({3H}-2DG) after prior loading or lactate dehydrogenase (LDH) enzyme release from the smooth muscle fragments . 3 . In alpha-toxin-permeabilized smooth muscle, but not in non-permeabilized muscle, GTP gamma S induced time- and concentration-dependent increases in labelled inositol phosphates . Carbachol (CCh) increased labelled inositol phosphates in both permeabilized and non-permeabilized muscle, although the increases were greater in non-permeabilized smooth muscle . The response to 100 microM CCh was severely reduced by 0.5 microM atropine . 4 . In permeabilized muscle the effects of GTP gamma S or CCh on inositol phosphate levels were reduced by treatment with pertussis toxin (PTX) and completely inhibited by GDP beta S . 5 . GTP gamma S caused a concentration-dependent inhibition of the CCh-induced increases in the levels of labelled inositol phosphates . Dibutyryl cyclic AMP or Sp-cAMPs (adenosine-3',5'-cyclic phosphorothiolate-Sp) reduced the effects of CCh on inositol phosphate levels . 6 . The results suggest that muscarinic AChR activation induces inositol phospholipid hydrolysis via more than one G-protein in this smooth muscle and that several mechanisms may contribute to the modulation of both stimulatory and inhibitory responses observed. J Bone Miner Res, 1995 May, 10(5), 726 - 34 Surface-associated proteins from Staphylococcus aureus demonstrate potent bone resorbing activity; Nair S et al.; Staphylococcus aureus infections are associated with rapid bone destruction in conditions such as osteomyelitis, bacterial arthritis, and infected orthopedic implant failure . How this bacterium induces bone destruction has not been defined . In studies of the role of oral Gram-negative bacteria in periodontal pathology, we have established that cell surface-associated proteins (SAPs) are potent stimulators of bone resorption . The surface-associated components from S . aureus have now been isolated and demonstrated to be extremely potent stimulators of bone resorption in the murine calvarial bone resorption assay . Bone resorption appears to be due to proteins, is not the result of contamination with lipoteichoic acid or muramyl dipeptide, and is potently inhibited by indomethacin and can be completely blocked by high concentrations of interleukin-1 receptor antagonist or TN3-19.12, a neutralizing monoclonal antibody to murine TNF . The SAP fraction can stimulate fibroblasts or monocytes to release osteolytic cytokines, but only at high concentrations . Fractionation of the SAPs by high performance liquid chromatography demonstrated that a number of fractions were osteolytically active . The most active contained a heterodimeric protein of molecular weight 32-36 kD . The presence of this osteolytically active surface-associated fraction may account for the bone resorption associated with local infection with S . aureus. Acta Paediatr, 1995 May, 84(5), 585 - 7 Multiple brain abscesses in a premature infant: complication of Staphylococcus aureus sepsis; Regev RH et al.; We report a premature infant with Staphylococcus aureus sepsis, most probably originating from an infected peripheral iv site, and complicated by multiple brain abscesses . Diagnosis was made by cranial ultrasonography and computed tomography . Since systemic antibiotic treatment failed, surgical drainage was performed . The same organism that caused the initial sepsis grew from the pyogenic material obtained . The child is currently severely handicapped. Antimicrob Agents Chemother, 1995 May, 39(5), 1173 - 7 Concentration-dependent synergy and antagonism between cefoperazone and imipenem against methicillin-resistant Staphylococcus aureus; Rand KH et al.; By the agar dilution checkerboard method, striking synergy between cefoperazone and imipenem was observed with 32 strains of methicillin-resistant Staphylococcus aureus (fractional inhibitory concentration indices, < or = 0.03 to 0.34; mean, 0.12 +/- 0.08) . By the double-disk diffusion method, the synergy was confirmed, but about 60% of strains showed antagonism manifest by truncation of the zone of inhibition around cefoperazone . The mechanism may involve auxiliary factors distinct from those essential for the expression of methicillin resistance. Antimicrob Agents Chemother, 1995 May, 39(5), 1134 - 9 In vivo verification of in vitro model of antibiotic treatment of device-related infection; Blaser J et al.; Device-related infections are difficult to treat with antibiotics alone . Standard susceptibility tests do not correlate with treatment success . Therefore, the utility of a pharmacokinetic in vitro model has been evaluated in comparison with the tissue-cage infection model in guinea pigs . The bactericidal activity of 28 treatment regimens has been studied by using three different test strains . In vitro efficacy was defined as reduction in the number of suspended or adherent bacteria, and in vivo efficacy was defined as reduction in the number of bacteria in tissue-cage fluid . Test results between the two models (in vivo and in vitro) correlated well, with correlation coefficients of 0.85 for in vivo efficacy versus in vitro efficacy against suspended bacteria and 0.72 for in vivo efficacy versus in vitro efficacy against adherent bacteria (P < 0.05) for Staphylococcus aureus, 0.96 and 0.82 (P < 0.05) for Staphylococcus epidermidis, and 0.89 and 0.97 for Escherichia coli, respectively . In contrast, standard susceptibility tests, ratios of MICs to trough or peak levels, ratios of the area under the curve to the MIC, or time above the MIC were not predictive for therapeutic outcome in either the in vitro or in vivo model . In both models, the bactericidal activity levels with combination regimens were significantly higher than those with single-drug regimens (P < 0.001) . Furthermore, rifampin combinations with either vancomycin, teicoplanin, fleroxacin, or ciprofloxacin were significantly more bactericidal against adherent bacteria than netilmicin combinations with vancomycin or daptomycin (P < 0.01) . Thus, in vivo verification of the pharmacokinetic in vitro model correlated well with the animal model . The in vitro model offers an alternative to ther animal model in experiments that screen and assess antibiotic regimens against device-related infections. J Biomed Mater Res, 1995 May, 29(5), 635 - 45 Contribution of vascular catheter material to the pathogenesis of infection: the enhanced risk of silicone in vivo; Sherertz RJ et al.; There is currently very little information to suggest that polymer materials used to make vascular catheters differ in their risk of infection . A rabbit model of subcutaneous Staphylococcus aureus infection was used to determine the relative risk of infection associated with silicone, polyurethane, polyvinylchloride, and Teflon catheters . Seven days after catheter implantation and inoculation with S . aureus, catheters were observed for gross purulence and quantitatively cultured . Silicone catheters were found to have a greater risk of grossly apparent infection (purulence) and a greater number of organisms removed from catheters by quantitative culture than the other three catheter materials (P < .01) . The risk of infection associated with silicone catheters decreased (P < .05) if the S . aureus inoculation was delayed for 2 days or if the catheters were preincubated in the subcutaneous space prior to insertion . The histology of the inflammatory response around the four catheter materials was evaluated at either 2 or 7 days after catheter insertion with or without S . aureus inoculation . Silicone catheters had greater associated inflammation (P < .05) with or without S . aureus inoculation . These results suggest that silicone catheter materials may have unique properties that increase the risk of infection after implantation . Further studies should be done to understand the mechanism(s) of these observations. Clin Infect Dis, 1995 May, 20(5), 1384 - 6 Gonococcal abscess of the obturator internal muscle: use of new diagnostic tools may eliminate the need for surgical intervention; Gurbani SG et al.; Obturator internal muscle (OIM) abscess or pyomyositis, a poorly recognized intrapelvic infection, is characterized by limping, pain in the hip with limited movement, and the sciatic type of radiating leg pain . With the use of advanced diagnostic tools such as computed tomography (CT), magnetic resonance imaging, and radionuclide scans, several cases of OIM abscess have been diagnosed; these cases have recently been reported in the English-language literature . Staphylococcus aureus is the predominant organism isolated from OIM abscesses . Gonococcal abscess involving the OIM has not been reported . We describe a case of gonococcal OIM abscess in an adolescent female whose condition was successfully diagnosed and managed with the use of CT-guided needle aspiration and antibiotic therapy . We review the English-language literature regarding the use of newly developed tools in the diagnosis of OIM abscess. J Clin Microbiol, 1995 May, 33(5), 1243 - 6 Evidence for the geographic spread of a methicillin-resistant Staphylococcus aureus clone between Portugal and Spain; Sanches IS et al.; Methicillin-resistant Staphylococcus aureus (MRSA) isolates collected during a 7-month period in 1992 and 1993 at Hospital Pulido Valente (340 beds), Lisbon, Portugal, were characterized by a combination of genotypic and phenotypic methods . Clonal identities were determined by probing ClaI digests (i) with a mecA probe and (ii) with a Tn554 probe and (iii) by pulsed-field gel electrophoresis (PFGE) patterns of chromosomal SmaI digests . mecA-ClaI type I was predominant among these isolates (38 of 43) . Most of these (37 of 38 {97.4%}) were associated with a single Tn554 pattern, pattern E, and the majority (23 of 38 {61%}) also showed a relatively uniform chromosomal background, as indicated by PFGE (PFGE pattern A) . The major clone (mecA-ClaI type I::Tn554 type E and PFGE pattern A) at Hospital Pulido Valente was indistinguishable by these molecular typing criteria from the dominant clone that had been identified in two major current outbreaks of MRSA disease in Spain (Barcelona and Madrid) . The Portuguese and Spanish clones also had a common heterogeneous class 3 phenotype and identical multidrug resistance patterns . The data presented in this work support the notion that MRSA clones can spread across considerable geographic distances. J Clin Microbiol, 1995 May, 33(5), 1121 - 8 Food-initiated outbreak of methicillin-resistant Staphylococcus aureus analyzed by pheno- and genotyping; Kluytmans J et al.; An outbreak of methicillin-resistant Staphylococcus aureus (MRSA) involving 27 patients and 14 health-care workers (HCW) was studied . The outbreak started in the hematology unit of the University Hospital Rotterdam, Dijkzigt, The Netherlands, and spread to the surgical unit . Twenty-one patients (77.8%) developed clinical disease, and five died . Subsequently, MRSA was detected in food and in the throat of one of the HCW who prepared food for hematology patients . Food contaminated by an HCW most likely caused the first case of MRSA septicemia . This route of transmission has not been described before . The outbreak strain was probably transmitted to the surgical unit by a colonized nurse, where it caused an explosive outbreak . Airborne probably transmitted to the surgical unit by a colonized nurse, where it caused an explosive outbreak . Airborne MRSA transmission played an important role in disseminating the organism . The outbreak was controlled within 6 months by intensifying surveillance, temporarily closing the affected wards, treating carriers, and instituting an MRSA ward outside the hospital . Phage typing, insertion sequence probing, protein A gene typing, and DNA fingerprinting by PCR revealed that all outbreak-related isolates were identical . By pulsed-field gel electrophoresis, all but one of the outbreak-related isolates were determined to be identical . Protein A gene typing identified numerous (11) repeat units in all outbreak-related isolates, which supports the suggestion that the outbreak strain may have been more virulent and more transmissible than other MRSA strains . Pheno- and genotyping studies underlined the value of DNA fingerprinting methods for investigation of MRSA epidemiology . Optimal discriminatory power was achieved by combining the results of four genotyping methods. Gaoxiong Yi Xue Ke Xue Za Zhi, 1995 May, 11(5), 295 - 9 Successful medical treatment for staphylococcal vertebral osteomyelitis complicated by spinal epidural abscess, psoas abscess and meningitis: a case report; Cheng TC et al.; A 42 year-old farmer was transferred to our hospital for recently exaggerated lower back pain . Neurological examination revealed an L4 radiculopathy on the right side . Meningitis developed after admission . MRI showed L4-5 osteomyelitis and discitis with contiguous spinal epidural abscess and right psoas abscess . Blood culture and CSF culture both grew Staphylococcus aureus . Because the patient refused to receive a drainage procedure, we gave him antibiotics which resulted in a favorable outcome. Kansenshogaku Zasshi, 1995 May, 69(5), 506 - 10 {DNA fingerprinting by arbitrarily primed polymerase chain reaction (AP-PCR) for methicillin-resistant Staphylococcus aureus}; Hojo S et al.; Recently, nosocomial outbreaks of MRSA have become an important social problem in Japan . To examine the routes of transmission of MRSA, the establishment of accurate MRSA typing system is essential . However, more recently, because MRSA strains with type II coagulase have been increasing, it is difficult to discriminate MRSA strains by the coagulase typing method . Under this background, our study was designed to evaluate the clinical significance of DNA fingerprinting by arbitrarily primed polymerase chain reaction (AP-PCR) . Several MRSA strains isolated from patients in our department were used in this study . To optimize the condition of AP-PCR, the differences of amplified products by AP-PCR were evaluated according to the following conditions: extracting methods of DNA from MRSA strains, buffer conditions, the temperature of AP-PCR, cycles of AP-PCR, and several primers . As a result, the optimal conditions of AP-PCR were as follows: extracted DNA using the InstaGene kit, amplified DNA by two-step AP-PCR using a M13 reverse primer with a buffer condition of 3.5 mM of magnesium chloride, and a pH of 8.5 . The results of AP-PCR correlated well with the results of pulsed-field gel electrophoresis . In conclusion, DNA fingerprinting by AP-PCR seems to be useful in examining the nosocomial MRSA outbreak. J Antimicrob Chemother, 1995 May, 35(5), 623 - 9 Azithromycin in an experimental Staphylococcus aureus abscess model; Bamberger DM et al.; We studied the efficacy and pharmacokinetics of azithromycin in a rabbit tissue-cage Staphylococcus aureus abscess model . A dosage of 15 mg/kg/day azithromycin was administered to rabbits with 24 h or 2 week old infected tissue cages and to uninfected controls . Concentrations of azithromycin were higher in the infected compared with the uninfected tissue cages . Azithromycin was effective in reducing the bacterial concentrations in both groups of infected tissue cages by approximately 3 log10 cfu/mL compared with untreated controls after 8 days of therapy . Fifty percent of the 24 h and 29% of the 2 week infected tissue cages became culture-negative. J Antimicrob Chemother, 1995 May, 35(5), 611 - 22 The influence of human granulocytes and monocytes on the antibacterial activity of cefetamet against Staphylococcus aureus; van den Broeck PJ et al.; The presence of human granulocytes and monocytes increased the activity of cefetamet against non-ingested Staphylococcus aureus . During ingestion of S . aureus by granulocytes, the activity of cefetamet was equal to that found in the absence of cells . Enhanced activity of cefetamet 30 mg/L was observed during ingestion of S . aureus by monocytes . The effect of cefetamet on S . aureus ingested by granulocytes was much less than that on bacteria in a liquid medium, the maximum growth inhibition (ERmax) being 0.15 and 0.90 log/h, respectively . The concentration at which 50% of the maximum effect was achieved (EC50) was 19.7 mg/L under both conditions . Cefetamet was also more active against S . aureus ingested by monocytes than against bacteria in medium (ERmax 0.97 and 0.77 log/h, respectively) . The EC50 under both conditions was 9.7 mg/L . The granulocyte-associated concentration of cefetamet was about on third of that in medium, whereas the monocyte-associated concentration was about two thirds of that in medium . The low intracellular activity of cefetamet against S . aureus ingested by granulocytes is explained primarily by the minimal intracellular growth of bacteria. Int J Immunopharmacol, 1995 May, 17(5), 445 - 52 The effect of carbamazepine and its reactive metabolite, 9-acridine carboxaldehyde, on immune cell function in vitro; Furst SM et al.; Carbamazepine, a widely used anticonvulsant, is associated with a wide range of adverse reactions including agranulocytosis, aplastic anemia and drug-induced lupus . It has also been reported to alter immune function in a variety of ways . We had previously demonstrated that carbamazepine is oxidized by activated neutrophils to several metabolites and this leads to covalent binding of the drug to the cells . It appears that the major metabolite responsible for this binding is 9-acridine carboxyaldehyde . In this study the effects on leukocyte function of carbamazepine and its leukocyte-generated metabolites were compared . Incubation of lymphocytes with 100 microM 9-acridine carboxaldehyde resulted in 40% cell death while carbamazepine at this concentration had no effect on viability . The effect on the immune cell function was investigated using the autologous mixed lymphocyte reaction (AMLR), allogeneic mixed lymphocyte reaction (MLR), lymphocyte transformation test (LTT) and mitogenesis assays . Alteration of immune cell function by the reactive metabolite, 9-acridine carboxyaldehyde, was demonstrated by an increased proliferation at low concentrations (0.08-1.0 microM) and inhibition at high concentrations (20-100 microM) in the allogeneic MLRs . Carbamazepine had no measurable effect . 9-Acridine appears to have more of an effect on B-cells since this augmentation-suppression phenomenon was also observed in mitogenesis assays with Staphylococcus aureus, a B-cell mitogen, in contrast to mostly inhibition observed in the mitogenesis assay with phytohemagglutinin, a T-cell mitogen . Again, carbamazepine had no measurable effects at comparable concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) J Clin Immunol, 1995 May, 15(3), 145 - 51 Staphylococcus aureus Cowan I-induced human immunoglobulin responses: preferential IgM rheumatoid factor production and VH3 mRNA expression by protein A-binding B cells; Kozlowski LM et al.; Protein A (PA), a cell wall component of SAC, activates human B cells by cross-linking the Fabs of membrane immunoglobulins . Recent data indicate that PA binds only to Fabs that use VH3 heavy chains, and thus it has been designated as a B-cell superantigen . We previously reported that Staphylococcus aureus Cowan I (SAC)-induced IgM rheumatoid factor (RF) by human PBMC was mediated by PA . Therefore, we sought to determine if SAC-induced IgMRF production was confined to PA-binding B cells and if these B cells were enriched for the expression of VH3 heavy chains . We observed that the elicitation of IgMRF in response to SAC was limited to a subset of B cells that bind PA and that this subset was enriched for VH3 mRNA expression . Taken together, these results suggest that IgMRFs produced in response to SAC are enriched for usage of VH3 heavy chains . Thus, this SAC-induced autoantibody response appears to represent a new B-cell superantigenic property of PA. East Afr Med J, 1995 May, 72(5), 322 - 4 Experience with ventriculo peritoneal shunts at the University of Nigeria Teaching Hospital, Enugu; Okoro BA et al.; In a study of 212 children at the University of Nigeria Teaching Hospital, Enugu who received ventriculo peritoneal shunt for hydrocephalus over a 13-year period (1977-1989, 14 had infected shunts and one developed shunt nephritis . Staphylococcus aureus was responsible for 36.4% of the positive cultures and was responsible for the only case of shunt nephritis . This is about the fourth case of shunt nephritis associated with Staphylococcus aureus shunt infection, in the literature . The pertinent features of this case were intraventricular haemorrhage as the cause of the hydrocephalus, frequent revision of shunts before the onset of nephritis and full recovery following intravenous antibiotic therapy and reimplantation of a new shunt. Int J Urol, 1995 May, 2(2), 17 - 23 Combined intraarterial cisplatin infusion and radiation therapy for invasive bladder cancer; Mizoguchi H et al.; Combined intraarterial cisplatin infusion and radiation therapy were performed as the initial treatment for 23 patients (mean age: 70 years) with invasive bladder cancers (T2 in 17, T3 in 6) who were suitable for total cystectomy . Of these patients, five who had multiple invasive cancers without laterality had their intrapelvic hemodynamics altered by embolizing a contralateral internal iliac artery . Cisplatin (50 mg) was infused into the internal iliac artery through a subcutaneous reservoir twice a week over three weeks while concurrent radiation therapy with 30 Gy, delivered in 15 fractions, was performed . Additional cisplatin infusions were given in six patients . After this combined therapy, total cystectomy and ileal conduit was performed in six patients and transurethral resection of bladder tumor (TURBT) in 17 . Two of the patients who underwent total cystectomy were found to exhibit a complete response . Therefore, the overall response rate was 87%, including 13 complete responses and seven partial responses . The complete response rates in patients with clinical stage T2 and T3 disease were 53 and 67%, respectively . The complete response rate was slightly higher in patients with a non-papillary cancer than in those with a papillary one . Toxic reactions included a decrease in bladder capacity in two patients and severe diarrhea due to methicillin-resistant Staphylococcus aureus colitis in one . Other forms of toxicity, including nausea, vomiting, neurotoxicity in the gluteal region, nephrotoxicity and myelosuppression, were tolerable . All but one of the patients are alive.(ABSTRACT TRUNCATED AT 250 WORDS) Br J Haematol, 1995 May, 90(1), 31 - 40 Administration of rHuGM-CSF activates monocyte reactive oxygen species secretion and adhesion molecule expression in vivo in patients following high-dose chemotherapy; Williams MA et al.; Microbicidal and cytocidal products of the respiratory burst and integrin adhesion molecule expression have been studied in monocytes from patients who received rHuGM-CSF during regeneration after high-dose chemotherapy . In this study, administration of rHuGM-CSF after high-dose chemotherapy significantly augmented the secretion of inducible products of the monocyte respiratory burst . Monocyte activation persisted for several weeks after the cessation of GM-CSF therapy . Under in vitro conditions that mimicked gram-negative (LPS) and gram-positive (opsonized Staphylococcus aureus) sepsis, the monocyte responded to such stimulation by exhibiting an enhanced release of hydrogen peroxide at both regeneration and several weeks later (P < 0.001) . Similarly, GM-CSF administration significantly augmented the phenotypic expression of the beta 2-integrin adhesion molecules and allowed the leucocyte-specific selectin, LAM-1, and the beta 2-integrins to respond normally to inflammatory stimulation by LPS . We further present evidence that GM-CSF therapy restored the otherwise refractory status of monocytes to inflammatory stimulation that existed in those patients given chemotherapy alone . The restoration of monocyte responsiveness by GM-CSF following high-dose chemotherapy could be a potentially valuable and hitherto not described action of rHuGM-CSF on monocyte function . We conclude that administration of GM-CSF may have the potential for restoring as well as augmenting the anti-microbial and anti-tumour function of the monocyte after high-dose chemotherapy. Blood, 1995 May 1, 85(9), 2482 - 9 Effect of granulocyte colony-stimulating factor treatment on ex vivo blood cytokine response in human volunteers; Hartung T et al.; We explored the ex vivo alteration in the cytokine release of stimulated blood taken from healthy volunteers treated subcutaneously with 480 micrograms granulocyte colony-stimulating factor (G-CSF) . In a double-blind, controlled, randomized study with 21 volunteers who received G-CSF once or twice 24 hours apart, we measured lipopolysaccharide (LPS)-inducible release of various cytokines and soluble receptors at different times after treatment . At day 1 after a single dose of G-CSF, mediator release was also initiated with muramyl dipeptide, Staphylococcus aureus enterotoxin A, lipoteichoic acid, streptolysin O, complement factor C5a, phytohemagglutinin, or phorbol myristate acetate . In blood from G-CSF-treated subjects, our major findings were (1) a maximal 12-fold increase in interleukin-1 receptor antagonist (IL-1ra) release and an increase of both the p55 and p75 soluble tumor necrosis factor (TNF) receptors; (2) a reduction in TNF release when using all the various stimuli described except LPS; (3) an increase in G-CSF and, to lesser extent, in IL-6, IL-8, and IL-10 release; and (4) an attenuation of interferon-gamma (IFN-gamma) and granulocyte-macrophage (GM)-CSF release . Our findings demonstrate that the major effect of G-CSF treatment is a change in the responsiveness of blood towards a variety of stimuli, which we interpret as a shift toward an antiinflammatory cytokine response. Blood, 1995 Apr 15, 85(8), 2202 - 11 Identification of signaling motifs within human Fc gamma RIIa and Fc gamma RIIb isoforms; Van den Herik-Oudijk IE et al.; To assess the functional capacity of the heterogeneous Fc gamma RII (CD32) family and to identify critical regions for functioning, we generated a panel of B-cell transfectants . The Fc gamma R-negative B-cell line IIA1.6 was transfected with wild-type or mutant human Fc gamma RIIa and IIb molecules . Solely Fc gamma RIIa-expressing IIA1.6 cells were capable of phagocytosing opsonized Staphylococcus aureus bacteria, and cross-linking of Fc gamma RIIa triggered a rapid induction of tyrosine phosphorylation after 20 seconds . Analysis of Fc gamma RIIa mutants identified the immunoreceptor tyrosine-based activation motif (ITAM; previously described as ARH-1 motif) within the IIa cytoplasmic tail to be critical for B-cell activation . In contrast, Fc gamma RIIb isoforms triggered tyrosine phosphorylation on cross-linking with much slower kinetics (> 3 minutes) than Fc gamma RIIa . Furthermore, solely Fc gamma RIIb molecules proved capable of downregulating {Ca2+}i and interleukin-2 production on co-cross-linking with sIgG in IIA1.6 . The Fc gamma RIIb-mediated functions were absent in Fc gamma RIIb mutants in which the tyrosine or leucine within the YSLL motif in a conserved 13-aa region (now known as immunoreceptor tyrosine-based inhibitor motif {ITIM}) were changed into phenylalanines . In conclusion, these data show the presence of functionally critical motifs within Fc gamma RII cytoplasmic tails . Fc gamma RIIa contains an ITAM involved in B-cell activatory functions, whereas the downregulatory activity of Fc gamma RIIb isoforms is linked to an ITIM. Cancer, 1995 Apr 15, 75(8), 2099 - 102 A phase II trial of extracorporeal plasma immunoadsorption of patient plasma with PROSORBA columns for treating metastatic breast cancer; Fennelly DW et al.; BACKGROUND . Circulating immune complexes (CIC) have been implicated as a cause of malignancy-associated immunosuppression and disease progression . Previous attempts to remove CIC by pheresis or immunoadsorption over a Staphylococcus aureus protein A column have resulted in a few clinical responses, however the relationship between removal of CIC and tumor response in these trials is not clear . Based on these data, a Phase II trial of immunoadsorption over a Staphylococcus aureus protein A column was initiated for patients with metastatic breast cancer . The authors sought to correlate clinical response with amount of CIC eluted from the columns after immunoadsorption . METHODS . The potential role of extracorporeal immunoadsorption was determined using protein A columns in treating patients with advanced breast cancer . An immunoadsorbent column composed of protein A was bound covalently to an inert silica matrix (PROSORBA {IMRE Corporation, Seattle, WA} column) . Patients underwent a 3-hour on-line procedure phlebotomizing 2000 ml of whole blood . Patient plasma was passed over PROSORBA columns to remove immunoglobulin G (IgG) and IgG-related CIC . The treated plasma then was reunited with formed elements and reinfused into the patient . Patients were treated three times per week for a total of 4 weeks . Analyses of tumor-associated Le(x)-containing CIC adsorbed on PROSORBA columns were performed using an enzyme-linked immunosorbent assay technique with a monoclonal antibody specific for the Le(x) moiety . RESULTS . Sixteen patients were entered in this Phase II study, with a mean age of 57 years (range, 40-69 years) . All patients received prior treatment for Stage IV breast cancer . The median number of PROSORBA treatments was 12 (range, 1-15 treatments) . No toxicities or major objective responses were seen noted the 16 patients . One patient with severe chest wall pain had a symptomatic response . The remaining patients all had disease progression . Analyses of column eluates from 11 patients in this study revealed no detectable Le(x)-containing immune complexes when compared with control subjects . CONCLUSIONS . Immunoadsorption over a Staphylococcus aureus Protein A column had no meaningful antitumor activity in patients with advanced breast cancer . In this cohort of patients, an elevated level of Le(x) CIC was not confirmed in the eluates of the column compared with a control group of patients without cancer. Pharmacoeconomics, 1995 May, 7(5), 416 - 27 Clinical and economic considerations in the use of third-generation oral cephalosporins; Chambers ST et al.; A number of oral third-generation cephalosporins (cefixime, cefetamet pivoxil, ceftibuten and cefpodoxime proxetil) have been widely trialled and are becoming available . In addition, cefdinir may also be marketed . Compared with first- and second-generation agents, the oral third-generation cephalosporins have an improved antibacterial spectrum and reduced minimum inhibitory concentrations against common Gram-negative pathogens . In contrast, with the exception of cefdinir, they are less active against Staphylococcus aureus . They have favourable pharmacokinetic profiles and are generally administered in once- or twice-daily regimens . They are well tolerated, but cefixime has been associated with a particularly high rate of diarrhoea . Possible clinical indications for the use of oral third-generation cephalosporins include upper and lower respiratory, genitourinary and soft-tissue infections and follow-on treatment of severe infections requiring hospitalisation . At present, these drugs offer no particular clinical advantages over standard therapy in most circumstances . However, they may be considered where there is hypersensitivity to penicillins, a high incidence of resistance to first-line therapy in the community, or failure of standard therapy . Further studies are needed to define the efficacy of oral third-generation agents in the prevention of rheumatic fever and as follow-on therapy for severe infections . The oral third-generation cephalosporins are generally more expensive than standard agents, but detailed studies that include extended costs (e.g . treatment of adverse effects, treatment of clinical failure, return visits to physicians) have yet to be reported. Science, 1995 Apr 7, 268(5207), 103 - 6 Structure of the cell wall anchor of surface proteins in Staphylococcus aureus; Schneewind O et al.; Many surface proteins are anchored to the cell wall of Gram-positive bacteria and are involved in the pathogenesis of these organisms . A hybrid molecule was designed that, when expressed in Staphylococcus aureus, was anchored to the cell wall and could be released by controlled enzymatic digestion . By a combination of molecular biology and mass spectrometry techniques, the structure of the cell wall anchor of surface proteins in S . aureus was revealed . After cleavage of surface proteins between threonine and glycine of the conserved LPXTG motif, the carboxyl of threonine is amide-linked to the free amino group of the pentaglycine crossbridge in the staphylococcal cell wall. J Biol Chem, 1995 Apr 7, 270(14), 8061 - 7 Identification of two 17-kDa rat parotid gland phosphoproteins, subjects for dephosphorylation upon beta-adrenergic stimulation, as destrin- and cofilin-like proteins; Kanamori T et al.; We previously reported that when 32Pi-loaded rat parotid slices are incubated with the beta-adrenergic agonist isoproterenol, the level of a soluble 32P-labeled 17-kDa protein (pp17) decreases rapidly (Kanamori, T., and Hayakawa, T . (1982) Biochem . Int . 4, 517-523) . Here we show that pp17 consists of two distinct phosphoproteins (pp17a and pp17b), identify their unphosphorylated forms (p17a and p17b, respectively), and provide evidence for their beta-adrenergic stimulation-induced dephosphorylation . Since p17a and p17b were predominant forms even in nonstimulated cells, peptides were generated from them with Staphylococcus aureus V8 protease or cyanogen bromide; subsequent sequencing of these peptides and homology search allowed identification of p17a and p17b as destrin- and cofilin-like proteins, respectively . Interestingly, they were also dephosphorylated in response to cholinergic stimulation . Because destrin and cofilin are actin-depolymerizing proteins whose activities are possibly regulated by their phosphorylation/dephosphorylation, the two parotid proteins reported here might be involved in cortical F-actin disruption observed in parallel with exocytotic amylase secretion. Bull Soc Pathol Exot, 1995 Apr, 88(5), 257 - 9 {Methicillin resistant Staphylococcus aureus . Decrease of incidence in a medical resuscitation unit at CHU Charles-Nicolle in Tunis}; Ben Hassen A et al.; Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen in hospitals . Between 1985 to 1988, there has been an increase in the number of MRSA (92 %) caused by an epidemic emergency in medical care unit of our hospital . Infections control measures allowed to stop these outbreaks (20 % MRSA in 1990) . But incidence of MRSA persist to decrease without supplement control measures and in 1994, only 7 % of Staphylococcus aureus strains are MRSA. Zentralbl Hyg Umweltmed, 1995 Apr, 197(1-3), 252 - 9 {Epidemiology and reasons for microbial resistance to biocides}; Kaulfers PM; In the last years the significance of microbial resistance to biocides like toxic heavy metal ions, aldehydes, phenoles, and cationic surface active agents has increased . However the genetical and functional mechanisms of the resistance to most of these compounds are still unknown . Till now it was possible to demonstrate a plasmid resistance in only some bacterial strains which were resistant to heavy metal ions, hexachlorophene, formaldehyde, benzalkoniumchloride, and chlorhexidine . In mercuric resistant Staphylococcus aureus and formaldehyde resistant E . coli strains the resistance genes are now well characterized . These strains are able to produce enzymes which degradade the disinfectant . The resistance to cationic surface active agents seems to depend on a modification of the bacterial cell wall . This is concluded from former investigations with benzalkoniumchloride resistant Pseudomonas strains, where it was possible to demonstrate different lipid contents in the cell wall from resistant and sensitive strains. Rev Latinoam Microbiol, 1995 Apr-Jun, 37(2), 127 - 34 {Recovery of Staphylococcus aureus after acid damage}; Assis EM et al.; The growth behavior of S . aureus in fresh cheese (Minas and Mozzarella) during their shelf life was studied in this research . The possibility of injury to this microorganism caused by increasing acidity was also investigated . Raw milk was inoculated with S . aureus FRIA-100 with approximately 10(6) cells/ml and cheese production was carried out according to normal procedures . They were stored at 7 degrees C during 40 days for Minas cheese and during 60 days for Mozzarella cheese . At 2 to 3 days intervals the following analyses were performed: acidity, pH, S . aureus count on Baird-Parker agar by traditional methods and by the method recommended by the American Public Health Association, to count repair of injured cells . We were certain of the presence of injured S . aureus when acidity was in the range of 0.7 to 0.8% expressed as lactic acid and when the count was 1.3 log higher. Rev Med Chil, 1995 Apr, 123(4), 457 - 63 {Chronic intermittent peritoneal dialysis: a 12-year experience}; Garcia R et al.; Since 1981, we have treated 46 adults aged 56 +/- 21 years, of whom 17 were diabetic, with intermittent peritoneal dialysis . Their mean time on dialysis has been 21 months with an accumulated experience of 493 months/patient . The program included 2 dialyses per week, 25 exchanges of 21 per session and 30 min of dwell time . Arterial pressure control has been satisfactory . Diabetic patients had lower levels of serum calcium, alkaline phosphatases and m-PTH . The incidence of peritonitis has been 1 episode/14 months/patient and the causative agent has been Staphylococcus aureus in 47% of episodes . Mean catheter duration has been 15 months and 1 episode/34 months/patient of exit site infection has been recorded and Staphylococcus aureus has been the causative agent in 83% of episodes . The risk of acquiring the first peritonitis was 12% at 3 months, 23% at 6 months and higher for non diabetic patients . Actuarial survival of treated patients at 12 and 24 months was 89 and 67% respectively . No differences in survival were recorded between diabetic and non diabetic patients . Fifty two percent of patients that dropped out continued on hemodialysis, 23% died, 11% abandoned treatment, 8% continued on chronic ambulatory peritoneal dialysis and 6% received a kidney allograft . We conclude that intermittent peritoneal dialysis is a good alternative treatment of chronic renal failure, even in diabetic patients. Infect Immun, 1995 Apr, 63(4), 1591 - 4 Possible receptor for exfoliative toxins produced by Staphylococcus hyicus and Staphylococcus aureus; Tanabe T et al.; Exfoliative toxin produced by Staphylococcus hyicus bound to the GM4-like glycolipid extracted from the skin of 1-day-old chickens but did not bind to glycolipid from adult chickens or suckling mice . Exfoliative toxin produced by Staphylococcus aureus bound to the GM4-like glycolipid extracted from the skin of suckling mice but not to glycolipid from 1-day-old or adult chickens . S . hyicus and S . aureus exfoliative toxins lost their toxicity by preincubation with GM4-like glycolipid from 1-day-old chickens and suckling mice, respectively. Infect Immun, 1995 Apr, 63(4), 1380 - 6 Capsular polysaccharide types 5 and 8 of Staphylococcus aureus bind specifically to human epithelial (KB) cells, endothelial cells, and monocytes and induce release of cytokines; Soell M et al.; In order to examine the possible implication of capsular polysaccharide (CP) types 5 and 8 (CP5 and CP8) from Staphylococcus aureus in the pathological mechanism associated with staphylococcal infections, we tested the immunomodulatory effects of CP5 and CP8 on human epithelial KB cells, endothelial cells, and monocytes . Using biotinylated CP5 and CP8, we provide evidence that both CPs bind to KB cells, endothelial cells, and monocytes in a dose- and calcium-dependent manner through specific interactions . These results were confirmed by competition experiments using soluble cell extracts . Furthermore, we show that CPs bind to identical cell membrane receptors on all three types of human cells and that human normal serum contains a factor(s) which inhibits the binding of both CPs to human KB cells, endothelial cells, and monocytes . The ability of CP5 and CP8 to stimulate the production of cytokines by the human cells was then examined . CP5 and CP8 trigger KB cells to produce interleukin-8 (IL-8); endothelial cells to produce IL-8 and IL-6; and monocytes to produce IL-8, IL-6, IL-1 beta, and tumor necrosis factor alpha . The release of cytokines by all three types of cells is time dependent and dose dependent, and the tumor necrosis factor alpha production by monocytes is not affected by the addition of polymyxin B . We further confirm that human normal serum inhibits the immunomodulatory effects of both polysaccharides on each kind of cell . These results confirm that S . aureus CPs act as bacterial adhesins having immunomodulatory effects for human cells. Infect Immun, 1995 Apr, 63(4), 1165 - 72 Endogenous gamma interferon, tumor necrosis factor, and interleukin-6 in Staphylococcus aureus infection in mice; Nakane A et al.; The production and roles of endogenous gamma interferon (IFN-gamma), tumor necrosis factor (TNF), and interleukin-6 (IL-6) in both lethal and nonlethal infections of Staphylococcus aureus were investigated in mice . In the case of nonlethal infection, although no bacteria were detected in the bloodstreams, bacteria that colonized and proliferated persistently for 3 weeks were found in the kidneys . All mice given lethal injections died within 7 days, and large numbers of bacteria were detected in the bloodstreams, spleens, and kidneys . The first peaks of IFN-gamma, TNF, and IL-6 were observed in the bloodstreams and spleens of the mice with nonlethal and lethal infections within 24 h . Thereafter, in the nonlethal cases, IFN-gamma, TNF, and IL-6 peaked again in the spleens and kidneys during the period of maximum growth of bacteria in the kidneys, although only IL-6 was detected in the sera . In contrast, in the case of lethal infection, the titers of IFN-gamma and IL-6 in the sera and TNF in the kidneys peaked before death . Effects of in vivo administration of monoclonal antibodies (MAbs) against IFN-gamma and TNF on the fates of S . aureus-infected mice were studied . In the nonlethal infections, anti-TNF alpha (anti-TNF-alpha) MAb-treated mice, but not anti-IFN-gamma MAb-treated mice, died as a result of worsening infection, suggesting that endogenous TNF plays a protective role in host resistance to S . aureus infection . In the mice that received lethal doses, injection of anti-TNF-alpha MAb accelerated death . However, although injection of anti-IFN-gamma MAb inhibited host resistance of the infected mice early in infection, most of the animals survived the lethal infection by injection of anti-IFN-gamma MAb, suggesting that endogenous IFN-gamma plays a detrimental role in S . aureus infection . Thus, this study demonstrated that IFN-gamma and TNF play different roles in S . aureus infection. Clin Infect Dis, 1995 Apr, 20(4), 895 - 9 Postoperative toxic shock syndrome; Graham DR et al.; We conducted a retrospective review of all cases of postoperative toxic shock syndrome (PTSS) occurring in two community hospitals from 1981-1993, during which time 390,000 surgical procedures were performed . The incidence was 0.003% (12 cases) . All wounds in these 12 cases, from those with scant superficial exudates to those with gross purulence, yielded Staphylococcus aureus . All tested isolates were susceptible to methicillin or cephalothin . Patients had a mean maximal temperature of 40 degrees C . All patients had a rash, most in a truncal, "sunburn" pattern . Eleven patients had desquamation . Mean time from surgery to onset of symptoms was 4 days . All patients required vigorous fluid resuscitation . No correlation could be demonstrated between the development of toxic shock syndrome and a patient's age, sex, preoperative skin preparation or administration of antibiotics, members of the surgical team, or duration of procedure . All patients with PTSS survived . PTSS should be considered in the differential diagnosis for the acutely febrile, systemically ill postoperative patient, even when surgical wounds are deceptively benign in appearance . Early recognition and treatment of PTSS is essential for successful outcome. Acta Paediatr Jpn, 1995 Apr, 37(2), 201 - 2 Transient low level of IgG3 induced by sepsis; Tabata N et al.; Staphylococcus aureus sepsis developed in a 14 year old girl . Immunological evaluation revealed low level of IgG3, although total IgG level was normal . The level of IgG3 increased gradually along with the recovery from sepsis . Immunoglobulin replacement therapy might have been useful in this patient, even though the total immunoglobulin level was within normal limits. Med Sci Sports Exerc, 1995 Apr, 27(4), 473 - 9 Osteomyelitis of the pubic symphysis in athletes: a case report and literature review; Karpos PA et al.; Groin pain is a common problem in athletes . Osteitis pubis, a chronic inflammatory condition involving the pubic symphysis, is a rare cause, and pyogenic osteomyelitis of the pubis is seen even more rarely in healthy athletes . We report one of four cases of pyogenic osteomyelitis of the pubis seen at our institution, review our experience with all four cases, and present a review of the literature (7 cases) . The diagnosis is established by the presence of extreme pain, point tenderness at the pubic symphysis, fever, and either a positive culture of blood, needle aspiration, or open biopsy of the pubis . White blood cell count, erythrocyte sedimentation rate, and the results of bone scan and computerized tomography may initially be normal and therefore cannot exclude the diagnosis . Prompt treatment with intravenous (i.v.) antibiotics effective against Staphylococcus aureus (causative organism in all documented cases-9/11) should initially be administered and then guided by culture and sensitivity information . Oral antibiotics should be given if the infection is responsive to i.v . antibiotic treatment . Prompt recognition and treatment with antibiotics may obviate the need for surgical debridement . All athletes who returned to sports activity did so by 6 months after diagnosis. Antimicrob Agents Chemother, 1995 Apr, 39(4), 982 - 4 Revised interpretation of oxacillin MICs for Staphylococcus epidermidis based on mecA detection; McDonald CL et al.; In 1992 and 1993, at The Ohio State University Medical Center, a larger proportion of Staphylococcus epidermidis strains required oxacillin MICs of 1 to 2 micrograms/ml than did Staphylococcus aureus strains . mecA genotype was correlated with antimicrobial susceptibility for selected clinical S . epidermidis strains . All 14 strains that required oxacillin MICs of < or = 0.25 microgram/ml and 2 of 5 strains that required oxacillin MICs of 0.5 microgram/ml were susceptible by 1-microgram oxacillin disk test and were mecA negative . Three of 5 strains that required oxacillin MICs of 0.5 microgram/ml and all 18 strains that required oxacillin MICs of > or = 1.0 microgram/ml were resistant by oxacillin disk test and were mecA positive . Current National Committee for Clinical Laboratory Standards MIC interpretive criteria may underestimate methicillin resistance among S . epidermidis strains. Jpn J Antibiot, 1995 Apr, 48(4), 563 - 70 Synergistic enhancement of in vitro antimicrobial activity of cefmetazole and cefotiam, cefamandole or cefoperazone in combination against methicillin-sensitive and -resistant Staphylococcus aureus . II . Effect of inoculum size; Uete T et al.; The inoculum effect was studied on the activity of cefmetazole and cefotiam, cefamandole or cefoperazone alone and in combination against 9 strains of methicillin-sensitive Staphylococcus aureus (MSSA) and 20 strains of methicillin-resistant Staphylococcus aureus (MRSA) by means of the checkerboard titration method with Mueller-Hinton agar plate using 10(6) and 10(8) CFU/ml . The antimicrobial activity against MSSA and MRSA was potentiated synergistically in combination of cefmetazole and these cephalosporins either with inoculum size 10(6) or 10(8) CFU/ml . At a concentration of cephalosporins < or = 6.25 micrograms/ml, the combination effect of cefmetazole and cefotiam or cefamandole against MRSA was more potent than that of cefmetazole and cefoperazone . With a higher inoculum size the effect was reduced . Under these conditions with a low dose level of drugs, the enhancement of the activity of cefmetazole by cefotiam was least influenced by inoculum size among cephalosporins studied. Jpn J Antibiot, 1995 Apr, 48(4), 553 - 62 Synergistic enhancement of in vitro antimicrobial activity of cefmetazole and cefazolin, cefotiam, cefamandole or cefoperazone in combination against methicillin-sensitive and -resistant Staphylococcus aureus . I . Effect of NaCl; Uete T et al.; The in vitro antimicrobial activity of cephamycin, e.g . cefmetazole and cephalosporin, such as cefazolin, cefotiam, cefamandole and cefoperazone, alone and in combination, was studied employing 9 strains of methicillin-sensitive Staphylococcus aureus (MSSA) and 30 strains of methicillin-resistant Staphylococcus aureus (MRSA) . Using the checkerboard agar dilution method, strong synergism was demonstrable in a majority of MSSA and MRSA strains for cefmetazole combined with these cephalosporins, with the minimum fractional inhibitory concentration index < or = 0.5 . In the presence of a concentration < or = 6.25 micrograms/ml of these cephalosporins in Mueller-Hinton agar medium, the activity of cefmetazole against MRSA was most prominently potentiated by cefotiam, followed by cefamandole, cefazolin and cefoperazone . At a concentration of 12.5 micrograms/ml, cefotiam and cefamandole showed a similar effect in potentiation of cefmetazole activity . In hypertonic agar medium containing 4% NaCl, these synergistic combination effects were reduced . However, the activity of cefmetazole and cefamandole in combination under these conditions was influenced to a lesser extent and more potent than that of other combinations. Microbiology, 1995 Apr, 141 ( Pt 4), 937 - 43 Variation in the size of the repeat region of the fibrinogen receptor (clumping factor) of Staphylococcus aureus strains; McDevitt D et al.; The ability of Staphylococcus aureus to bind to fibrinogen and fibrin is believed to be an important factor in the initiation of foreign body and wound infections . Recently, the gene encoding the fibrinogen receptor (clumping factor, ClfA) of S . aureus strain Newman was cloned and sequenced . The ClfA protein possesses a highly unusual 308 residue dipeptide repeat region composed predominantly of Asp and Ser . Polymerase chain reaction analysis of seven different strains showed that the size of the clfA repeat coding region varies from 580 bp to 1320 bp . In contrast, the clfA region A is the same size in each strain . The size of the clfA repeat region did not correlate with the ability of these strains to form clumps in a solution of fibrinogen . Indeed, the strain with the smallest repeat size of 580 bp clumped almost as well as strain Newman . Each strain of S . aureus examined contained several high molecular mass proteins that reacted with anti-ClfA region A antibody . In some cases the molecular mass of the major protein varied in accordance with the length of the coding sequence for the repeat region R. FEMS Microbiol Lett, 1995 Apr 1, 127(3), 195 - 9 Influence of iron depletion on growth kinetics, siderophore production, and protein expression of Staphylococcus aureus; Trivier D et al.; Growth rates, siderophore secretion, and bacterial proteins of two clinical isolates of Staphylococcus aureus were studied over 72 h of growth in iron-supplemented and iron-restricted chemically defined media . Under iron restriction the growth rates were decreased to different extents depending on the strain . Production of siderophore was detected in the mid-exponential and stationary phases of growth . The expression of iron-regulated proteins of 81, 23, and 17 kDa was time-dependent, associated with the same stage of growth, and might be involved in siderophore efficiency. FEMS Microbiol Lett, 1995 Apr 1, 127(3), 165 - 70 Interaction of the chromosomal Tn 551 with two thermosensitive derivatives, pS1 and p delta D, of the plasmid pI9789 in Staphylococcus aureus; Sohail M et al.; The plasmid pI9789::Tn552 carries genes conferring resistance to penicillins and to cadmium, mercury and arsenate ions . The presence of Tn551 at one location in the chromosome of Staphylococcus aureus enhances the frequency of suppression of thermosensitivity of replication of the plasmids pS1 and p delta D which are derivatives of pI9789::Tn552 . Bacteriophage propagated on the bacteria in which thermosensitivity of replication had been suppressed was used to transduce cadmium resistance to S . aureus PS80N . The cadmium-resistant transductants obtained carried plasmid pS1 or p delta D with a copy of Tn551 inserted into a specific site on pS1 but into several different sites on p delta D . The possible mechanisms of the suppression are discussed. Kansenshogaku Zasshi, 1995 Apr, 69(4), 408 - 12 {Restriction fragment length polymorphism of coagulase gene of methicillin-resistant Staphylococcus aureus can be used as an epidemiological marker}; Tobita M et al.; In order to see the restriction fragment length polymorphism (RFLP) of coagulase gene of Methicillin-resistant Staphylococcus aureus (MRSA) and the type of coagulase, a total of 118 strains of MRSA from 1986 to 1994 was collected from the Akita University Hospital, six hospitals in Akita Prefecture and a hospital in Aomori Prefecture . The RFLP analysis using the Alu I restriction enzyme brought us four kinds of electrophoretic patterns (A, B, C, D) . Patterns A (87%) and B (13%) were detected in our hospital and only pattern A was detected in other hospitals in Akita . On the other hand, Patterns A (68%), B (26%), C (3%) and D (3%) were detected in Aomori . In coagulase type analysis, all strains in Akita showed type II, and the strains in Aomori also showed type II except for one strain (type IV) . From the above results, the polymorphism of coagulase gene of MRSA can be useful as an epidemiological marker to examine the extent of MRSA infection in a prefecture unit or in a larger area. Free Radic Biol Med, 1995 Apr, 18(4), 633 - 9 Modification of neutrophil oxidant production with diphenyleneiodonium and its effect on bacterial killing; Hampton MB et al.; Diphenyleneiodonium (DPI), an inhibitor of the NADPH oxidase, has been used to distinguish between oxidative and nonoxidative killing of Staphylococcus aureus and Escherichia coli by neutrophils . The rate of killing of S . aureus was inhibited by 77% in the presence of 10 microM DPI, compared to 81% measured under anaerobic conditions . DPI represents a convenient and accessible alternative to an anaerobic environment or using neutrophils from patients with chronic granulomatous disease, for eliminating oxidative killing . The killing of E . coli was also inhibited by DPI . The effect was more apparent at 30 min than at 10 min, suggesting that E . coli can be killed rapidly by nonoxidative mechanisms that become less efficient at later times . DPI was used at concentrations less than 10 microM to determine how this affected production of the three major neutrophil oxidants, superoxide, hydrogen peroxide, and hypochlorous acid, and to determine the effect of partial inhibition of oxidant production on the killing of S . aureus . Unexpectedly, lower concentrations of DPI (0.1-2 microM) inhibited hydrogen peroxide and hypochlorous acid production 10-30% more than they inhibited superoxide production . Correlation of hydrogen peroxide or hypochlorous acid production with the killing of S . aureus showed that up to 30% inhibition had no effect on the rate of killing, implying that agents that impair neutrophil oxidant production less than this will not compromise bacterial killing . Higher inhibition of oxidant production led to a linear decline in the rate of killing. Appl Environ Microbiol, 1995 Apr, 61(4), 1438 - 43 Identification and purification of a new staphylococcal enterotoxin, H; Su YC et al.; A staphylococcal enterotoxin which elicited an emetic response in monkeys but did not share antigenic determinants with any of the identified enterotoxins was identified and purified from Staphylococcus aureus FRI-569 . The emetic activity of this new enterotoxin was neutralized only by antibodies specific to it and not by antibodies to enterotoxins A, B, C, D, and E or toxic shock syndrome toxin 1 . Immunodiffusion assays did not detect cross-reactivity between this new and all the other identified enterotoxins . The purification procedure involved removal of the enterotoxin from culture supernatant fluids by batch adsorption with CG-50 resin, CM-Sepharose FL ion-exchange chromatography, and Sephacryl 100 HR and Bio-Gel P-30 gel filtration . The molecular weight of this enterotoxin, 27,300, determined by gel filtration on Sephacryl 100 HR agreed with the molecular weight, 28,500, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) . The apparent migration of this enterotoxin determined by SDS-PAGE did not shift in the presence of a disulfide reducing agent, indicating that it is composed of a single-chain protein . The N-terminal amino acid sequence of the enterotoxin was determined to be Glu-Asp-Leu-His-Asp-Lys-Ser-Glu-Leu-Thr-Asp-Leu-Ala-Leu-Ala-Asn-Ala-Tyr- Gly- Gln-Tyr-Asn-His-Pro-Phe-Ile-Lys-Glu-Asn-Ile, which did not match the N-terminal sequences of any known proteins . The isoelectric point of the enterotoxin determined by isoelectric focusing was about 5.7.(ABSTRACT TRUNCATED AT 250 WORDS) Arch Biochem Biophys, 1995 Apr 1, 318(1), 89 - 96 Primary structure of a coagulant enzyme, bilineobin, from Agkistrodon bilineatus venom; Nikai T et al.; The amino acid sequence and disulfide bridge location of the coagulant enzyme, named bilineobin, isolated from the venom of Agkistrodon bilineatus was determined by Edman sequencing of the peptides derived from digests with cyanogen bromide, clostripain, Staphylococcus aureus V8 protease, trypsin, and chymotrypsin . This enzyme has a molecular weight of 57,000 Da by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; however, bilineobin consists of 235 amino acids and has a calculated molecular weight of 26,481 . The enzyme contains fucose, GlcNAc, galactose, mannose and NeuAc and six N-linked glycosylation consensus sites . The carboxyterminal amino acid, proline, was determined using carboxypeptidase Y . The six disulfide bonds of bilineobin link Cys78 to Cys234, Cys120 to Cys188, Cys178 to Cys203, Cys7 to Cys141, Cys152 to Cys167, and Cys28 to Cys44 . The amino acid sequence similarity to flavoxobin (T.C . Shieh et al., 1988, J . Biochem (Tokyo) 103, 596-605) and batroxobin (N . Itoh et al., 1987, J . Biol . Chem . 262, 3132-3135) was 67% . The deglycosylated enzyme more rapidly generated fibrinopeptide A than native bilineobin. Tijdschr Diergeneeskd, 1995 Apr 1, 120(7), 208 - 13 {Subclinical and clinical mastitis on dairy farms in The Netherlands: epidemiological developments}; Schukken YH et al.; In this review a number of the main epidemiological developments in the field of udder health in the Netherlands are summarized . The changes considering the bulk milk somatic cell count are described, as are the population dynamics of subclinical mastitis . Additionally, clinical mastitis caused by Escherichia coli and Staphylococcus aureus and their risk factors are reviewed . Finally, some future developments and the possible applications of these developments are discussed. J Nutr, 1995 Apr, 125(4), 817 - 22 A mixture of nucleosides and nucleotides increases bone marrow cell and peripheral neutrophil number in mice infected with methicillin-resistant Staphylococcus aureus; Matsumoto Y et al.; We studied the effects of a mixture of nucleosides and nucleotides on the peripheral neutrophil number and the proliferation of bone marrow cells in mice challenged with methicillin-resistant Staphylococcus aureus . BALB/c mice were fed a nucleotide-free 20% casein diet (control) or this diet supplemented with nucleosides and nucleotides orally (Expt . 1) or intraperitoneally (Expt . 2 and 3) . On d 10, the mice were challenged intravenously with methicillin-resistant S . aureus (6.7 x 10(12) colony forming units/L) . In Expt . 1 and 2, numbers of total and differential counts of blood leucocytes were counted on d 0 (before), 1, 3 and 5 after the infection . In Expt . 3, 30 min before killing, bromodeoxyuridine (20 mg/kg), an analogue of thymidine, was administered intraperitoneally and its incorporation in the DNA synthetic phase of bone marrow cells was determined at 0 h (before), 3, 6 and 24 h after the infection . Mice fed the supplemented diet had higher (P < 0.05) leucocyte and neutrophil numbers on d 0 compared with the control group . The neutrophil numbers tended to be greater in the supplemented group at 1, 3 and 5 d after the infection . Intraperitoneal administration of nucleosides and nucleotides increased (P < 0.05) neutrophil numbers before and after the infection . Twenty-four h after the infection, incorporation of bromodeoxyuridine into the DNA synthetic phase of bone marrow cells in the group administered nucleosides and nucleotides was higher (P < 0.05) than in the control group . We conclude that, following methicillin-resistant S . aureus injection, intraperitoneal administration of a nucleoside-nucleotide mixture may stimulate bone marrow cell proliferation and increase the peripheral blood neutrophil numbers . Oral administration may elicit weaker effects. J Med Microbiol, 1995 Apr, 42(4), 237 - 45 Epidemiological data on Staphylococcus aureus strains producing synergohymenotropic toxins; Prevost G et al.; DNA hybridisation of 309 consecutive Staphylococcus aureus clinical isolates with oligonucleotide probes specific for genes encoding Panton-Valentine leucocidin (luk-PV) and gamma-haemolysin (hlg) revealed that 99% of randomly selected strains carried the hlg locus whereas only 2% harboured the luk-PV as well as the hlg loci . Only 1% of the strains did not possess either gene . In a clinical prospective study of independent S . aureus strains, 58 Panton-Valentine leucocidin (PVL)-producing isolates were shown to be responsible for primary skin infections, mainly furuncles (86%) . Phage susceptibility patterns and pulsed field gel electrophoresis (PFGE) profiles of DNA were shown to be polymorphic epidemiological markers of PVL-producing strains . In eight patients with recurrent furuncles, the PVL-producing strains isolated either from furuncles or from the anterior nares were considered to be identical in each based upon phage sensitivity profiles or PFGE patterns. J Infect Dis, 1995 Apr, 171(4), 897 - 902 Ampicillin, sulbactam, and rifampin combination treatment of experimental methicillin-resistant Staphylococcus aureus endocarditis in rabbits; Chambers HF et al.; Ampicillin or amoxicillin at 625-800 mg/kg/day, in combination with a beta-lactamase inhibitor, each is as effective as vancomycin in animal models of methicillin-resistant Staphylococcus aureus endocarditis . Studies were done to determine whether the addition of rifampin would permit lowering the dose of ampicillin into the range recommended for use in humans without loss of efficacy . The efficacy of ampicillin/sulbactam (300/150 or 150/75 mg/kg/day intramuscularly, in three divided doses) in combination with rifampin (5 mg/kg intramuscularly, three times daily) was compared with that of vancomycin (25 mg/kg intravenously, twice daily, or 30 mg/kg intramuscularly, three times daily) in the rabbit model of methicillin-resistant S . aureus aortic valve endocarditis . Neither ampicillin/sulbactam nor rifampin alone was effective . The ampicillin/sulbactam/rifampin regimen was as effective as vancomycin . This regimen may be an alternative to vancomycin in treatment of methicillin-resistant S . aureus infections. Transfusion, 1995 Apr, 35(4), 298 - 302 Comparison of bacteria growth in single and pooled platelet concentrates after deliberate inoculation and storage; Wagner SJ et al.; BACKGROUND: The ability to store pools of platelet concentrates (PCs) for extended periods would provide logistical flexibility . However, reports of severe adverse reactions due to the transfusion of contaminated PCs led to an examination of whether the total bacteria levels after storage of pools containing a deliberately inoculated platelet unit would be significantly different than the levels in paired unpooled concentrates . STUDY DESIGN AND METHODS: A single PC was deliberately inoculated on Day 0 with one of three bacterial species (0.1-8.0 colony-forming units/mL) . On Day 1, the deliberately inoculated PC was divided into three equal parts and either 1) pooled with 5 half-volume, ABO- and Rh-identical PCs; 2) similarly pooled and white cell reduced; or 3) kept as a control . Sterile connections were used during pooling; modified storage containers were used to ensure the correct surface-to-volume ratio of the single unit . RESULTS: Between Day 2 and Day 5 of storage, in 26 of 36 paired samples, nonfiltered pools containing Escherichia coli had greater total numbers of bacteria than did the paired single PCs . Day 2 pools had total bacteria levels approximately five times higher (colony-forming units/mL x container volume) than those in single units (p < 0.05) . There was rapid growth of Staphylococcus aureus by Day 2 in pooled and unpooled PCs; by Day 3, total bacteria levels were approximately five times higher in pools than in single units (p < 0.05) . Between Days 3 and 5 of storage, in 23 of 27 paired samples, nonfiltered pools containing S . aureus had greater total bacteria levels than the single PCs . By Day 5, 15 of 16 non-white-cell reduced pools had total levels of Staphylococcus epidermidis bacteria approximately five times those in the paired single PCs . Greater total bacteria levels in pooled units than in single units generally occurred when bacteria in pools reached the stationary phase of growth (when bacteria concentration became constant), and they were well correlated with the sixfold volume of pooled units . White cell reduction did not substantially affect the time required to attain stationary phase . CONCLUSION: The potential during storage for greater total bacteria levels in pools than in single PCs is a consequence of the greater volume of the pool. Vet Immunol Immunopathol, 1995 Apr, 45(3-4), 211 - 20 Immortalization and characterization of bovine peritoneal macrophages transfected with SV40 plasmid DNA; Stabel JR et al.; A transformed bovine peritoneal macrophage cell line was developed and characterized . Primary peritoneal macrophages were transformed by calcium-phosphate transfection with SV40 plasmid DNA . The transformed cell line retained the morphology of resident peritoneal macrophages as determined by light microscopy and histochemical analysis showed non-specific esterase activity . In addition, immunohistochemical staining of transformed peritoneal macrophages for lysozyme activity was positive . Transformed cells phagocytized Staphylococcus aureus, lysed chicken red blood cell (RBC) targets with and without opsonization and produced hydrogen peroxide radicals and interleukin-6 upon stimulation with opsonized zymosan and lipopolysaccharide, respectively . Transformed cells were also able to ingest and kill Mycobacterium paratuberculosis, an acid-fast bacillus . These results suggest that this cell line should be useful to study interactions between the bovine and intracellular pathogens. J Hosp Infect, 1995 Apr, 29(4), 265 - 73 Methicillin-resistant Staphylococcus aureus (MRSA) isolated in clinics and hospitals in the Fukuoka city area; Kono K et al.; Bacteriological and clinical studies were carried out on 280 strains of Staphylococcus aureus isolated in clinics and hospitals in the Fukuoka city area from September 1990 to March 1991 . The percentage of methicillin-resistant S . aureus (MRSA) strains studied was 41.4% (116 of 280) . Of 116 MRSA strains, 48 (41.3%) produced coagulase VII and 21 (18.1%) produced coagulase II . The mean age of the patients who harboured MRSA, 70.5 +/- 16.9 years, was significantly higher than that of patients with methicillin-sensitive S . aureus (MSSA), 44.2 +/- 29.3 years (P < 0.001) . MRSA was detected more frequently than MSSA in sputum (P < 0.01), while MSSA was detected more frequently than MRSA in pus (P < 0.01) . Ninety (89.1%) of 101 strains of MRSA were isolated from inpatients and 98 (71.0%) of 130 strains of MSSA were isolated from outpatients . Similar number of MRSA strains were recovered in a variety of hospitals, indicating that there was no relationship between hospital size (number of beds) and the incidence of MRSA . As for drug susceptibility, coagulase VII-producing MRSA strains were more sensitive to clidamycin (P < 0.01) and more resistant to minocycline (P < 0.01) than were other MRSA strains. Eur J Clin Microbiol Infect Dis, 1995 Apr, 14(4), 282 - 90 Variation and persistence of methicillin-resistant Staphylococcus aureus strains among individual patients over extended periods of time; Maslow JN et al.; To determine the strain variation and persistence among isolates of methicillin-resistant Staphylococcus aureus (MRSA) cultured from patients with colonization over extended time spans, pulsed-field gel electrophoresis was used to analyze the isolates from 47 patients for whom at least two mecA-positive isolates collected a minimum of six months apart were available . For 22 (47%) patients, the isolates represented multiple distinct strains of Staphylococcus aureus, while 20 (43%) patients had only a single strain detected; five (11%) patients had similar, genetically related isolates . MRSA were frequently associated with mucocutaneous abnormalities; 29 (62%) patients had focal cutaneous defects, and ten (21%) had chronic dermatitis . Multiple strains of MRSA were detected more frequently than single strains among patients in whom the initial focus of MRSA resolved clinically and another mucocutaneous defect subsequently developed compared to patients with clinically persistent foci (11/15 versus 9/23, respectively; p = 0.05, Fisher's exact test) . Among the 21 patients in this series for whom isolates cultured within a two-month time span were available, there were seven (33%) patients with multiple strains of MRSA, including one patient with polyclonal bacteremia . In summary, patients with long-term MRSA colonization often have several different strains of MRSA, which typically change overtime in association with removal or resolution of a colonized focus and the recurrence of mucocutaneous defects. Can J Vet Res, 1995 Apr, 59(2), 124 - 8 Prevalence of coagulase gene polymorphism in Staphylococcus aureus isolates causing bovine mastitis; Aarestrup FM et al.; This study was conducted to investigate polymorphism of the coagulase gene of Staphylococcus aureus causing bovine mastitis . One hundred eighty-seven strains of S . aureus were isolated from bovine mastitic milk samples obtained from 187 different Danish dairy farms . The isolates were characterised for restriction fragment length polymorphism (RFLP) of the coagulase gene . A variable region of the coagulase gene was amplified using the polymerase chain reaction (PCR) followed by AluI restriction enzyme digestion . A total of 15 different RFLP patterns were observed . The predominant pattern was found in 35% of the isolates . The ease of analysing coagulase gene polymorphisms among a large number of strains, and the multiple distinct polymorphic patterns generated, supports the use of this technique in epidemiological investigations of bovine mastitis . The predominating variants may have predelection for causing intramammary infections. Minerva Cardioangiol, 1995 Apr, 43(4), 155 - 9 {Splenic abscess after angiography . Considerations on a clinical case}; Tridico F et al.; A case of splenic abscess in a diabetic, cardiopathic and arteriopathic 49-year-old man is reported . The abscess developed on a previous splenic infarction caused by an embolus coming from an intraventricular thrombus . The angiographic procedure was the source of bacterial contamination (Staphylococcus aureus) . The patient was successfully treated with splenectomy. Am J Infect Control, 1995 Apr, 23(2), 87 - 94 Recommendations for preventing the spread of vancomycin resistance: recommendations of the Hospital Infection Control Practices Advisory Committee (HICPAC); Conjunctivitis in a long-term care facility; Section of Geriatric Medicine, Riverview Health Centre, Winnipeg, Manitoba, CanadaOBJECTIVE: To determine the prevalence, incidence, and etiology of conjunctivitis in residents of a long-term care facility . DESIGN: Prospective surveillance of episodes of conjunctivitis during two 6-month periods . SETTING: Three-building, 319-bed, multipurpose, long-term care facility including units for personal care, chronic care, palliative care, rehabilitation, respiratory rehabilitation, and chronic ventilator patients . RESULTS: The incidence of conjunctivitis on different units varied from 0.6 to 3.5 per 1,000 patient-days . One building had a significantly higher rate of acute conjunctivitis and, within that facility, the most highly impaired patients had significantly more disease . Patients with chronic conjunctivitis generally were admitted with conjunctivitis; institutionally acquired conjunctivitis proceeding to chronic conjunctivitis seldom was observed except on the chronic ventilator unit . Residents with chronic conjunctivitis were significantly more likely to have a diagnosis of glaucoma, ectropion, or entropion . Potential bacterial pathogens were isolated from 3 (21%) of 14 and 24 (38%) of 69 acute episodes for which cultures were available in the two study periods . Staphylococcus aureus was the most frequent pathogen isolated . Empiric therapy was usually with topical sodium sulamyd or gentamicin, with substantial interphysician variability in prescribing patterns . Symptom duration did not differ for episodes which did or did not receive topical antimicrobials . CONCLUSION: Conjunctivitis is common in this facility and occurs with increased frequency in more highly impaired residents . Most episodes may not be due to bacterial infection . Further study of optimal management approaches to conjunctivitis is necessary. Naunyn Schmiedebergs Arch Pharmacol, 1995 Apr, 351(4), 329 - 36 Analysis of receptor-G protein interactions in permeabilized cells; Wieland T et al.; Receptor-induced binding of the stable GTP analogue, guanosine 5'-{gamma-thio}triphosphate (GTP {gamma S}), to guanine nucleotide-binding regulatory proteins (G proteins) was measured in various permeabilized cells . In myeloid differentiated human leukemia (HL-60) cells, permeabilized with either digitonin, streptolysin O or Staphylococcus aureus alpha-toxin, binding of GTP{gamma S} induced by three distinct chemoattractant receptors was observed . The extent of receptor-stimulated GTP{gamma S} binding (maximally about 2-fold) was independent of the type of permeabilizing agent used . In human erythroleukemia cells permeabilized with digitonin, agonist activation of thrombin and neuropeptide Y receptors increased GTP{gamma S} binding by 1.8- and 1.5-fold, respectively . Finally, in adherently grown human embryonic kidney cells permeabilized with digitonin, activation of the stably expressed human muscarinic m3 receptor increased GTP{gamma S} binding by about 1.6-fold . In digitonin-permeabilized HL-60 cells, a quantitative analysis of formyl peptide receptors and interacting G proteins was performed . About 50,000 formyl peptide receptors per cell were detected . Agonist binding to these receptors was fully sensitive to regulation by guanine nucleotides and pertussis toxin . The number of high-affinity GTP{gamma S} binding sites, most likely representing heterotrimeric G proteins, was calculated to be about 670,000 per cell . Stimulation of formyl peptide receptors led to the activation of about 130,000 of high-affinity GTP{gamma S} binding sites, indicating a ratio of about three activated G proteins per one agonist-activated receptor.(ABSTRACT TRUNCATED AT 250 WORDS) J Antimicrob Chemother, 1995 Apr, 35(4), 473 - 81 Binding affinities of beta-lactam antibodies for penicillin-binding protein 2' in methicillin-resistant staphylococcus aureus; Sumita Y et al.; We devised an accurate procedure with which to measure the affinities of beta-lactam antibiotics for penicillin-binding protein (PBP) 2' in methicillin-resistant Staphylococcus aureus (MRSA) . In the present study, we used two isogenic strains of MRSA, on heterogeneous and the other homogeneous, derived from the methicillin-susceptible strain FDA209P, harbouring the mecA gene . In these MRSA strains, PBP2' was saturated by {14C}benzylpenicillin (PCG) at a concentration of 300 mg/L . In addition, the saturation of PBP2' by {14C}PCG required an incubation period of 30 min . According to these results, the precise affinities of beta-lactam antibiotics for PBP2' were determined by the 'accurate competition assay', using a high concentration of {14C}PCG and extending the reaction time . This procedure yielded lower IC50 values of beta-lactams than the 'usual competition assay' . However, each beta-lactam had almost the same affinity for PBP2' in heterogeneous and homogeneous strains . These results suggest there is a factor(s) other than PBP2' responsible for controlling resistance levels and the heterogeneity or homogeneity of MRSA strains. Ther Drug Monit, 1995 Apr, 17(2), 107 - 12 Changes in the serum protein binding of vancomycin in patients with methicillin-resistant Staphylococcus aureus infection: the role of serum alpha 1-acid glycoprotein levels; Morita K et al.; The relationship between albumin or alpha 1-acid glycoprotein (AAG) levels and vancomycin (VCM) protein binding was studied in 44 serum samples from the 10 patients with methicillin-resistant Staphylococcus aureus (MRSA) infection receiving VCM therapy . Eighty serum samples from 80 healthy subjects were used as a control for albumin and AAG levels . The protein binding percentage of VCM in the serum of the patients varied widely from 27 to 62% . The mean albumin level (34 g/L) was significantly lower than that in healthy subjects (46 g/L) . There was no correlation between the binding percentage and serum albumin level in the patients (r = -0.25, p > 0.1) . The mean AAG level in the patients (1.51 g/L) was significantly higher than in healthy subjects (0.59 g/L) . There was a significant correlation between the binding percentage of VCM and serum AAG level (r = 0.63, p < 0.001) . The binding percentage of VCM individual patients changed in parallel to serum AAG and C-reactive protein (CRP) levels, which is a useful marker of acute phase response . There were also significant correlations between AAG, albumin, and CRP levels . The present results indicate that the increased AAG level in serum of the patients appeared to have a significant effect on the protein-binding characteristics of VCM. Eur J Vasc Endovasc Surg, 1995 Apr, 9(3), 314 - 8 Rifampicin impregnated Dacron grafts: no development of rifampicin resistance in an animal model; Sardelic F et al.; AIM: Rifampicin impregnated Dacron grafts have been shown to be effective at preventing vascular graft infection in different animal models . The development of resistance to rifampicin would be a major drawback to the widespread use of such a graft . We aimed to determine how readily this would occur by using a sheep animal model . METHODS: Under general anaesthetic a 2cm long, 5mm diameter Dacron interposition graft inpregnated with 1.2 mg/ml rifampicin was placed in the left carotid artery . An extreme challenge of methicillin resistant Staphylococcus aureus (MRSA) using an inoculum of 10(9) colony forming units was placed directly onto the graft . The grafts were harvested at 3 weeks and cultures of the graft and tissues were taken . The presence or absence of any abscess formation, anastomotic disruption and graft thrombosis was noted . Any positive growths were identified and if found to be the same as the inoculum, the bacteria were used as the inoculum for another sheep . This was repeated once more . Thus we started with three sheep initially and used a total of nine sheep . RESULTS: There were no deaths . All grafts were infected with the same MRSA strain, confirmed on phage typing . There were three abscess and one anastomotic disruption . Seven of the grafts were occluded . The minimal inhibitory concentration (MIC) of the infecting inoculum and the bacteria retrieved were determined using the agar dilutional method . The MIC for the three initial inocula was < 0.007 mg/l . All subsequent strains isolated had an MIC of < 0.015 mg/l . This was a difference of one dilution and not significant . CONCLUSION: There was no development of rifampicin resistance using this animal model. Acta Med Okayama, 1995 Apr, 49(2), 81 - 9 Current status of antimicrobial susceptibility in MRSA isolates typed by coagulase and phage typing in Okinawa; Lotsu DK et al.; The incidence of nosocomial infections with methicillin-resistant Staphylococcus aureus is of great concern in Japan and the developed world as a whole . Simple typing techniques like coagulase and phage typing are quick and useful for monitoring and evaluating these organisms . In view of this, the current status of antimicrobial susceptibility in Staphylococcus aureus (S . aureus) isolates in Okinawa typed by coagulase and phage typing was studied . Of 508 isolates, methicillin-resistant S . aureus (MRSA) comprised 54.3% (minimum inhibitory concentration (MIC) > or = 16 micrograms/ml) . Coagulase type II and phage group III were the most prevalent, comprising 65.2% and 38%, respectively . These were followed by phage non-typable group and coagulase type III with 36.6% and 12.7%, respectively . Compared to a previous study conducted in 1989, there has been an increase of about 17% in the MRSA isolation rate with a concomitant increase of about 11% in the coagulase type II MRSA isolation rate and a decrease of about 27% in the isolation rate of coagulase type III MRSA . Using a panel of 16 antibiotics, coagulase type II MRSA were resistant to all except Arbekacin and Vancomycin . Arbekacin and Vancomycin were the sole antibiotics to which resistance was not expressed by any of the isolates . With regard to the methicillin-sensitive S . aureus (MSSA), coagulase type III and phase group III were the most prevalent, comprising 25.9% and 32.3%, respectively. Nippon Kyobu Geka Gakkai Zasshi, 1995 Apr, 43(4), 493 - 6 {A case of infective mitral endocarditis after percutaneous transvenous mitral commissurotomy}; Inoue M et al.; We encountered a case of infective mitral endocarditis (IE) due to Methicillin resistant staphylococcus aureus (MRSA) after percutaneous transvenous mitral commissurotomy (PTMC) . A 65-year-old woman underwent PTMC for mitral stenosis . Two days later, she incurred a fever of more than 40 degrees C, and MRSA was detected in her blood culture . Inflammation and renal function deteriorated despite transvenous administration of antibiotics . Transesophageal echocardiogram revealed vegetation at the mitral valve . Mitral valve replacement was performed after diagnosis of IE, and MRSA was identified from the vegetation . The patient improved with the transvenous application of antibiotics including vancomycin hydrochloride . The blood culture became negative and inflammation was suppressed . In view of this case, we consider that IE is a possible complication following PTMC. Eur J Pediatr, 1995 Apr, 154(4), 273 - 4 Thigh pain due to obturator internus phlegmon: a diagnostic challenge; Godfroid N et al.; A 9-year-old boy with thigh pain, high fever, hyperleukocytosis and positive blood cultures for Staphylococcus aureus had a left obturator internus phlegmon . Lack of evidence for hip infection or osteomyelitis in a child with thigh pain, high fever and hyperleukocytosis points to a possible infection of pelvic structures . Abscess or phlegmon of the obturator internus muscle is a very rare condition . The most frequent agent is Staphylococcus aureus . CT of the pelvis is the procedure of choice to show diffuse swelling of the muscle or collection of pus . Conservative treatment with adequate antibiotics is the treatment of choice. Lab Anim, 1995 Apr, 29(2), 177 - 9 Granulomatous dermatitis and mastitis in two SPF rats associated with a slowly growing Staphylococcus aureus--a case report; Kunstyr I et al.; An isolated occurrence of multifocal severe granulomatous dermatitis and mastitis accompanied by extensive calcifications is described in 2 female Sprague-Dawley, SPF breeding rats . Poorly growing Staphylococcus aureus of uncertain lysotype (probably lysotype II) was isolated from the lesions of both rats . The source of infection could not be determined . No further cases in the closed barrier maintained breeding colony occurred in the following 7 months . Difficulties in interpreting these findings and the practical consequences relating to the hygienic status of the barrier breeding colony are discussed. J Virol Methods, 1995 Apr, 52(3), 265 - 72 A combined staphylococcal coagglutination assay for rapid identification of rotavirus and adenovirus (COARA); Peret TC et al.; An improved staphylococcal coagglutination test was developed for rapid detection, in a single assay, of rotavirus and adenovirus in stool samples (COARA) . Suspensions of Staphylococcus aureus coated respectively with anti-rotavirus and anti-adenovirus sera were used to identify these viruses in 327 stool samples of children . The samples were also tested by an enzyme immunoassay . The data analysis has demonstrated a high degree of correlation between the two assays. Electrophoresis, 1995 Apr, 16(4), 595 - 603 Charge and size effects in the capillary zone electrophoresis of nuclease A and its variants; Kalman F et al.; The migration behavior of nuclease A from Staphylococcus aureus and 11 of its variants in capillary zone electrophoresis (CZE) was investigated in the light of their three-dimensional structure known from X-ray crystallography and nuclear magnetic resonance (NMR) measurements . Nuclease A (molecular mass 16.8 kDa, pKa 10.3) and the variants differ only in a single amino acid residue and have a very similar crystal structure . With the use of coated quartz capillaries and suitable buffers, the protein migration was investigated at pH from 2.8 to 9.5 without interference by wall adsorption . Although the selectivity of the electrophoretic system for the proteins was mainly determined by their charge differences, certain variants having the same net charge could also be readily separated under nondenaturing conditions . For instance, the mobility of variant K116A was sufficiently higher than that of K116G so that they could be separated by CZE . The structures of both variants are the same except for the solvent-exposed loop containing residue 116 . For this reason, the difference in electrophoretic mobilities can be attributed to the fact that in K116G the backbone of the 112 to 117 amino acids protrudes slightly from the protein, with a concomitant increase in the hydrodynamic radius with respect to that of K116A . Consequently, K116G shows a smaller mobility than K116A due to its larger hydrodynamic radius despite its smaller molecular mass . The interpretation of the experimentally measured mobilities of such closely related proteins therefore requires not only consideration of their electrostatic charge but also the fine details of their molecular structures. Toxicon, 1995 Apr, 33(4), 437 - 49 Ammodytoxin A acceptor in bovine brain synaptic membranes; Krizaj I et al.; Ammodytoxin A, the presynaptic neurotoxin from Vipera ammodytes ammodytes venom, was found to bind specifically and with high affinity to bovine cortex synaptic membrane preparation . The detected ammodytoxin A high-affinity binding was characterized by equilibrium binding analysis which revealed a single high-affinity binding site with Kd 4.13 nM and Bmax 6.67 pmoles/mg of membrane protein . 125I-ammodytoxin A was covalently cross-linked to its neuronal acceptor using a chemical cross-linking technique . As revealed by subsequent SDS-PAGE analysis and autoradiography, 125I-ammodytoxin A specifically attached to membrane components with apparent mol . wts 53,000-56,000 . Besides by the native ammodytoxin A, the binding of radioiodinated ammodytoxin A to the neuronal acceptor was highly attenuated, also by other two iso-neurotoxins from V . a . ammodytes venom, ammodytoxins B and C, and neurotoxin crotoxin B from the venom of the South American rattlesnake (Crotalus durissus terrificus) . Vipera berus berus phospholipase A2 was a weaker inhibitor, whereas nontoxic phospholipase A2, ammodytoxin I2 and myotoxic phospholipase A2 homologue, ammodytin L, both from V . a . ammodytes venom as well, were very weak inhibitors . No inhibitory effect on 125I-ammodytoxin A specific binding at all was, however, obtained with alpha-dendrotoxin, beta-bungarotoxin and crotoxin A, respectively . Treatment of synaptic membranes with proteinase K and Staphylococcus aureus V-8 proteinase, a combination of PNGase F and neuroaminidase, heat or acid lowered the 125I-ammodytoxin A specific binding to various extents but never completely abolished it . The ammodytoxin A binding site in bovine synaptic membranes is thus most likely a combination of membrane glycoprotein acceptor and membrane phospholipids . As ammodytoxin A reduced the second negative component of the perineural waveform, measured on mouse triangularis sterni preparation, which is very likely a result of an inhibition of a fraction of the terminal K+ currents, the ammodytoxin A acceptor could well be connected with K+ channels. Pathol Biol (Paris), 1995 Apr, 43(4), 364 - 9 {Penetration of ceftriaxone (1 or 2 gr intravenous) into mediastinal and cardiac tissues in man}; Viviand X et al.; Ceftriaxone penetration into heart tissues (valves, myocardium, auricles and pericardium) and mediastinal tissues (fat and sternal bone) was evaluated after two regimens of ceftriaxone administration . Ten patients were given 1,000 g intravenously of ceftriaxone 30 min . before anesthesia . Ten other patients received the same dose and then a second 1,000 mg dose at the time of initiation of cardiopulmonary bypass . Similar and very satisfactory ceftriaxone tissue penetrations were observed in both groups . However, for some patients in the two groups and mainly in the sternal bone at the time of thorax closure, ceftriaxone levels in tissues were less than the MICs for some potential pathogens (Methicillin susceptible Staphylococcus aureus and Staphylococcus epidermidis) . During the different steps of the surgical procedures all patients in both groups had tissue levels greater than the MICs for Gram negative aerobic bacilli, except for Pseudomonas spp. Pathol Biol (Paris), 1995 Apr, 43(4), 329 - 35 {Dynamics of the nasal colonization by methicillin resistant Staphylococcus aureus in patients hospitalized in an intensive care unit}; Mounier M et al.; Prospective study on MRSA nasal cariage was done during two months in an intensive care unit in Limoges University Hospital . Nasal swab specimens were taken daily and cultured on selective and non selective media . Sixty eight patients were included in this study (878 swab collected, 575 MRSA isolated) . Patients mean age was 62 years and stay period mean was 12.3 days (median: 7 days) . Among these patients, 16 were already carrying MRSA when entering in the unit and 26 became positive for MRSA during their stay . The mean colonisation delay was 5.5 days (median 4 days) . All patients, except one, have shown a nasal carriage during all their stay . During the study period, 17 patients became infected and only two patients neither carried nasal MRSA before and during infection . In all cases, glycopeptide treatment did not affect nasal carriage . Colonisation and infection risk factors were discussed. J Electron Microsc (Tokyo), 1995 Apr, 44(2), 66 - 71 Ultrastructural localization of penicillin-binding sites in Staphylococcus aureus using penicillin G-BSA-gold; Morioka H et al.; Penicillin G (PcG) conjugated to bovine serum albumin (BSA) was found to retain antibacterial activity against Staphylococcus aureus as judged by growth inhibition and by ultrastructural analyses of the PcG-BSA-treated bacterium . Using the PcG-BSA conjugated to colloidal gold, binding sites of PcG were localized in S . aureus with electron microscopy using a postembedding labeling technique . To our surprise, labeling was observed both on the cell wall and in the cytoplasm of S . aureus . A part of the labeling observed may be attributed to penicillin-binding proteins (PBPs) or their precursors . Also there may be other molecules which bind with PcG in the cell wall and cytoplasm of S . aureus. Immunology, 1995 Apr, 84(4), 645 - 52 Activation of effector functions by immune complexes of mouse IgG2a with isotype-specific autoantibodies; Rajnavolgyi E et al.; Analysis of five monoclonal autoantibodies, rheumatoid factors produced by hybridomas generated from spleen cells of BALB/c mice repeatedly infected with A/PR/8/34 human influenza A virus, revealed that they recognized distinct but spatially related epitopes . The differing isoallotypic specificity of the IgM and IgA monoclonal antibodies correlated with the presence of Ile258 and Ala305, respectively . Although these data suggest that the epitopes recognized are within the CH2 domain, all antibodies failed to inhibit IgG antigen reactivity with Staphylococcus aureus protein A (SpA), C1q, mouse C3, human Fc gamma RI or mouse Fc gamma RII, activities known to be predominantly determined by CH2 domain structures . Reactivity of the IgA antibody, Z34, with IgG2b allowed further specificity studies using a panel of 26 mutant IgG2b proteins, each having single amino acid replacements over the surface of the CH2 domain . The only substitution that affected Z34 reactivity was Asn/Ala297, which destroyed the glycosylation sequon, resulting in secretion of an aglycosylated IgG molecule . The epitope recognized by Z34 therefore seems to be located outside of the Fc gamma R and C1q binding sites, but to be dependent on the presence of carbohydrate for expression . In contrast to the binding studies, complement activation by aggregated IgG2a, through classical or alternative pathways, was inhibited by the presence of autoantibodies . The functional significance of isotype-specific autoantibody in immune regulation is discussed. Br J Haematol, 1995 Apr, 89(4), 771 - 9 Absence of G-CSF receptors and absent response to G-CSF in childhood Burkitt's lymphoma and B-ALL cells; Tomeczkowski J et al.; The expression of the granulocyte colony-stimulating factor (G-CSF) receptor in childhood Burkitt's lymphoma (BL) cells, and the mitogenic effect of G-CSF on these cells, was studied in a panel of 13 Epstein-Barr virus (EBV) positive and negative BL cell lines derived from nine children . G-CSF receptor mRNA expression was investigated by Northern blot analysis and reverse transcriptase polymerase chain reaction (RT-PCR) . Binding of G-CSF to BL cell lines was measured by chemical crosslinking of 125I-G-CSF, and proliferation by thymidine incorporation . Inducibility of the G-CSF receptor was studied by stimulation with interleukin-1 beta, tumour necrosis factor-alpha, Staphylococcus aureus Cowan A, anti-human IgM, phorbol myristate acetate, calcium ionophore A23187, and by infection in vitro by immortalizing and non-immortalizing strains of EBV . BL cell lines, unstimulated or stimulated by biological reagents or EBV infection, did not bind radioionated G-CSF in crosslinking experiments . No stimulation by recombinant human G-CSF was observed in 3H-thymidine incorporation assays . No G-CSF receptor mRNA was detected by Northern blot analysis or RT-PCR in BL cell lines . It is concluded that G-CSF plays no direct stimulatory role in the growth of these malignant B-cells, making a deleterious influence of G-CSF in the clinical treatment situation unlikely. Appl Microbiol Biotechnol, 1995 Apr, 43(1), 65 - 9 Molecular cloning and expression in Escherichia coli of bleomycin-resistance gene from a methicillin-resistant Staphylococcus aureus and its association with IS431 mec; Bhuiyan MZ et al.; A gene that confers bleomycin resistance was cloned from the chromosomal DNA of methicillin-resistant Staphylococcus aureus (MRSA) B-26 into the plasmid pUC18 . It is of chromosomal origin rather than plasmid and exists in the chromosome making a cluster with the kanamycin-resistance gene . We found that the nucleotide sequence of the bleomycin-resistance gene from the chromosome of MRSA B-26 is identical to that from a staphylococcal plasmid, pUB110 . The partial sequence of IS431mec was also found upstream from the DNA fragment containing the bleomycin- and kanamycin-resistance genes.
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