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 phag