FEMS Microbiol Lett, 1993 Jan 15, 106(2), 147 - 55 Serotype F double- and triple-converting phage insertionally inactivate the Staphylococcus aureus beta-toxin determinant by a common molecular mechanism; Carroll JD et al.; The precise molecular mechanism of Staphylococcus aureus beta-toxin inactivation by the serotype F triple-converting phage phi 42, phi A1 and phi A3 was investigated . Sequence analysis of the phi 42 (attP) and Staphylococcus aureus (attB) attachment sites and the left (attL) and right (attR) chromosomal/bacteriophage DNA junctions of individual lysogens, each harbouring a triple-converting phage, revealed the presence of a common 14-bp core sequence in all four sites . These findings indicate that the genomes of the triple-converting phage integrate into the 5'-end of the beta-toxin gene (hlb) by a site- and orientation-specific mechanism identical to that previously described for the serotype F double-converting phage phi 13. FEMS Microbiol Lett, 1993 Jan 15, 106(2), 123 - 7 Cloning of the replication region on the bacteriocinogenic plasmid pRJ9 of Staphylococcus aureus; de Oliveira SS et al.; A 5.8-kb ClaI fragment of pRJ9, a bacteriocinogenic plasmid of Staphylococcus aureus, was cloned in the unique ClaI site of pRJ5 . The recombinant plasmid obtained, pRJ23, failed to confer bacteriocin production and immunity to bacteriocin on host cells . The cloned fragment was shown to contain the complete replicon of pRJ9 . Attempts to clone the 4.4-kb ClaI fragment of pRJ9 were unsuccessful, apparently due to the inactivation of the basic replicon of the cloning vector . Therefore, plasmid pRJ5 cut at its ClaI site appears to be a suitable vector for cloning replication regions of plasmids that can replicate in S . aureus. Gene, 1993 Jan 15, 123(1), 99 - 103 The use of degenerate, sensor gene-specific, oligodeoxyribonucleotide primers to amplify DNA fragments from Staphylococcus aureus; Bayles KW; The sensor proteins of bacterial two-component regulatory systems comprise a large family of proteins that are involved in environmental sensing and signal transduction . To study these proteins in the Gram+ pathogen, Staphylococcus aureus, two pairs of degenerate oligodeoxyribonucleotides (oligos) that corresponded to conserved sequences contained within sensor protein-encoding genes were synthesized . Using these oligo primers, DNA fragments from S . aureus were amplified by polymerase chain reaction (PCR), cloned in Escherichia coli, and sequenced . Comparison of the deduced amino acid sequences from these cloned fragments to the sequences contained in the GenBank database suggest that some of the PCR products were derived from sensor protein-encoding genes . However, several other fragments were identified that encoded peptides with up to 65% identity to transport proteins . Given the biochemical and functional properties of some of these proteins, these data suggest that sensor and transport proteins may be evolutionarily related. Blood, 1993 Jan 15, 81(2), 454 - 61 Subtractive cDNA cloning of a novel member of the Ig gene superfamily expressed at high levels in activated B lymphocytes; Kozlow EJ et al.; Using subtractive cDNA cloning we have isolated a series of cDNA clones that are exclusively or selectively expressed in B lymphocytes . mRNA transcripts from one such cDNA clone, referred to as BL11, were found to be expressed at low levels in RNA from normal B lymphocytes, but at very high levels in RNA from in vitro activated B lymphocytes . One major 2.5-kb BL11 mRNA transcript was detected, while low levels of 4.8-, 1.8-, and 1.6-kb transcripts were also found . BL11 mRNA transcripts were absent or present at low levels in RNA prepared from resting or mitogen activated T cells, a variety of lymphoid cell lines including several B-cell lines, and several different tissues . Low levels of BL11 transcripts were found in poly(A) RNA purified from brain and lung . A study of the kinetics of BL11 mRNA accumulation in B lymphocytes stimulated in vitro with Staphylococcus aureus Cowan strain I showed a rapid induction of BL11 mRNA within 2 hours of stimulation with peak expression by 16 hours and a mild decrease with time following the peak levels . Consistent with the in vitro data, in situ hybridization using antisense BL11 RNA probes and human tonsillar tissue localized BL11 transcripts in B-cell-enriched areas . Multiple BL11 cDNA and genomic clones were isolated and sequenced to complete and verify the BL11 cDNA sequence (2,404 bp) . A 615-nucleotide open reading frame predicted to encode for a 205-amino acid protein with a molecular weight of 23 Kd was identified . Search of protein data bases with the predicted BL11 protein showed homologies to several members of the Ig superfamily . Analysis of the predicted protein showed a likely signal peptide, a single membrane spanning region, and one V-like Ig domain with three predicted n-glycosylation sites . Southern blot analysis of human genomic DNA suggested that BL11 is a single copy gene without evidence of rearrangement . Primer extension and S1 nuclease mapping identified four tightly clustered transcriptional start sites approximately 40 bp upstream of the predicted translation start site . The first 270 bp of the promoter region were sequenced and found to contain a CATAA box rather than a TATAA box and several DNA motifs found in activation genes . BL11 should prove to be an interesting gene that likely encodes for a protein involved in B-cell activation. Med Clin (Barc), 1993 Jan 9, 100(1), 9 - 13 {Relationship between nosocomial infection and hospital mortality . Multicenter study}; Rodriguez-Rumayor G et al.; BACKGROUND: Hospital mortality related to infections acquired in the hospital setting has not been well studied in Spain . We carried out a study of seven hospitals in order to assess and quantify the problem . METHODS: The study period included three months of observations (between November 1, 1989 and January 31, 1990), and data pertaining to all deaths of patients hospitalized for a minimum of 24 hours were collected . The number of people admitted within the study period was 16,025, and the number of deaths registered and included in our study was 488 (3%) . The data were obtained from the patient's medical history one week after death as well as from the hospital physicians on the case . In order to quantify the interobserver variability derived from the classification criteria, the simple kappa index was calculated and averaged to form an ordinal scale . RESULTS: 216 (44.3%) of 488 deaths included in our study had no infection, 138 (28%) had an infection no-hospital-associated, and 134 (27%) had nosocomial infection (50-10%--"causally related to death", 59-12%--"contributing to death", and 25-5%--"not related to death") . The lower respiratory tract infections, bacteremias and surgical wound infections were the most related to cause of death . Staphylococcus aureus was the pathogen most frequently associated with the infections found at the time of death . CONCLUSIONS: Those patients admitted with non-fatal diseases made up the greatest percentage (39.9%) of deaths from nosocomial infections . The infection was considered the direct cause of death in 18.8% of these cases, although the differences found had no statistical significance. Science, 1993 Jan 8, 259(5092), 227 - 30 Evidence for a clonal origin of methicillin resistance in Staphylococcus aureus; Kreiswirth B et al.; Soon after methicillin was introduced into clinical practice in the early 1960s, resistant strains of Staphylococcus aureus (MRSA) appeared, bearing a newly acquired resistance gene, mecA, that encodes a penicillin binding protein, PBP2a . MRSA have spread throughout the world, and an investigation of the clonality of 472 isolates by DNA hybridization was performed . All 472 isolates could be divided into six temporally ordered mecA hybridization patterns, and three of these were subdivided by the chromomosomal transposon Tn554 . Each Tn554 pattern occurred in association with one and only one mecA pattern, suggesting that mecA divergence preceded the acquisition of Tn554 in all cases and therefore that mecA may have been acquired just once by S . aureus. J Immunol Methods, 1993 Jan 4, 157(1-2), 225 - 31 Comparative analysis of using MTT and XTT in colorimetric assays for quantitating bovine neutrophil bactericidal activity; Stevens MG et al.; Two different tetrazolium compounds were compared for use in a colorimetric assay for quantitating bovine neutrophil bactericidal activity against Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, and Brucella abortus . The tetrazolium compounds tested included 3-{4,5-dimethylthiazol-2-yl}-2,5-diphenyltetrazolium bromide (MTT) and sodium 3,3'-{1{(phenylamino)carbonyl}-3,4- tetrazolium}-bis(4-methoxy-6-nitro) benzene sulfonic acid hydrate (XTT) . The MTT and XTT colorimetric bactericidal assays were conducted by incubating antibody-opsonized bacteria with neutrophils in microtiter plates for 30 and 60 min at ratios of ten and 100 bacteria per neutrophil . Neutrophils were then lysed with saponin and samples were incubated 30 min with MTT or XTT plus coenzyme Q (CQ) . Dead bacteria and lysed neutrophils did not react with MTT or XTT plus CQ . Live bacteria converted XTT to water soluble orange formazan in the presence of CQ and MTT to insoluble purple formazan . Absorption of formazan produced by bacteria from XTT was measured at 450 nm . Formazan produced by bacteria from MTT was solubilized by adding isopropanol and measured by absorption at 560 nm . Absorption of both types of formazan was directly related to viable bacteria cell number and used to determine the number of bacteria not killed by neutrophils . The percentage of bacteria killed by neutrophils was determined by extrapolation from a standard formazan curve that was derived by incubating MTT or XTT plus CQ with known numbers of bacteria . The XTT and MTT colorimetric bactericidal assays produced comparable results when used to measure bovine neutrophil bactericidal activity against S . aureus, E . coli, L . monocytogenes, and B . abortus . However, the assay using XTT was quicker and easier to perform because bacteria converted XTT to a formazan that did not need to be solubilized before measuring absorption. Bull Hosp Jt Dis, 1993-95, 53(4), 7 - 9 Septicemia in a child undergoing callotasis limb lengthening; Minty I et al.; We report a patient undergoing femoral callotasis lengthening using the dynamic axial fixator to correct a post-infective leg length discrepancy of 7.8 cm . Seventeen days after the operation, the patient developed a pin site infection, which was successfully treated by oral antibiotics . On the 34th post-operative day, the infection reoccurred, and was accompanied by generalized malaise, vomiting and pyrexia . Serology identified Staphylococcus aureus enterotoxin . Following removal of the fixator, the child recovered, but only four cm of lengthening was achieved . The pins probably acted as a persistent foreign body, with local inflammation creating favorable ground for bacterial infection . The role of the previous multifocal osteomyelitis unclear, but it could have acted as a continuous source of pathogens . The resulting toxemia was not immediately suspected, and could have resulted in the loss of the patient had the fixator not been removed promptly. Vojnosanit Pregl, 1993 Jan-Feb, 50(1), 39 - 42 {Analysis of bacterial flora in burn patients}; Tomanovic B et al.; Bacterial flora of burn wound smears and hemocultures of 25 patients with burns have been followed up and analysed in the period of one year . Pseudomonas auruginosa have been most commonly isolated from burn wound smears, in 406 (5.6%) and Staphylococcus aureus, in 371 (47.14) samples . The most important hemoculture isolates has been Staphylococcus aureus found in 34 (57.6%) of samples . One multiresistant strain of Staphylococcus aureus represented 91% of all Staphylococcus aureus strains isolated from hemocultures and 84% from burn wound smears . Serologic and pyocin typing of Pseudomonas auruginosa proved the presence of several epidemic strains. Biochem Mol Biol Int, 1993 Jan, 29(1), 149 - 59 Comparative study of the properties of plasmid mediated staphylococcal beta-lactamase as expressed in Escherichia coli; Saha D et al.; An unmodified natural plasmid of Staphylococcal origin having beta-lactamase determinant was introduced by transformation into Gram-negative Escherichia coli HB101 recA-SmR (plasmidless) . Beta-lactamase of Staphylococcus aureus is extracellular . In recipient transformed E . coli HB101, the enzyme shows periplasmic location . The enzyme is expressed well in E . coli, and the basal level of enzyme production in E . coli, is approximately 3-4 fold higher than that of S . aureus . Beta-lactamase is inducible both in S . aureus and E . coli . Enzymatic properties as indicated by substrate profile and Km values are found to be similar. J Physiol, 1993 Jan, 460, 247 - 71 The effects of MgADP on cross-bridge kinetics: a laser flash photolysis study of guinea-pig smooth muscle; Nishiye E et al.; 1 . The effects of MgADP on cross-bridge kinetics were investigated using laser flash photolysis of caged ATP (P3-1(2-nitrophenyl) ethyladenosine 5'-triphosphate), in guinea-pig portal vein smooth muscle permeabilized with Staphylococcus aureus alpha-toxin . Isometric tension and in-phase stiffness transitions from rigor state were monitored upon photolysis of caged ATP . The estimated concentration of ATP released from caged ATP by high-pressure liquid chromatography (HPLC) was 1.3 mM . 2 . The time course of relaxation initiated by photolysis of caged ATP in the absence of Ca2+ was well fitted during the initial 200 ms by two exponential functions with time constants of, respectively, tau 1 = 34 ms and tau 2 = 1.2 s and relative amplitudes of 0.14 and 0.86 . Multiple exponential functions were needed to fit longer intervals; the half-time of the overall relaxation was 0.8 s . The second order rate constant for cross-bridge detachment by ATP, estimated from the rate of initial relaxation, was 0.4-2.3 x 10(4) M-1 s-1 . 3 . MgADP dose dependently reduced both the relative amplitude of the first component and the rate constant of the second component of relaxation . Conversely, treatment of muscles with apyrase, to deplete endogenous ADP, increased the relative amplitude of the first component . In the presence of MgADP, in-phase stiffness decreased during force maintenance, suggesting that the force per cross-bridge increased . The apparent dissociation constant (Kd) of MgADP for the cross-bridge binding site, estimated from its concentration-dependent effect on the relative amplitude of the first component, was 1.3 microM . This affinity is much higher than the previously reported values (50-300 microM for smooth muscle; 18-400 microM for skeletal muscle; 7-10 microM for cardiac muscle) . It is possible that the high affinity reflects the properties of a state generated during the co-operative reattachment cycle, rather than that of the rigor bridge . 4 . The rate constant of MgADP release from cross-bridges, estimated from its concentration-dependent effect on the rate constant of the second (tau 2) component, was 0.35-7.7 s-1 . To the extent that reattachment of cross-bridges could slow relaxation even during the initial 200 ms, this rate constant may be an underestimate . 5 . Inorganic phosphate (Pi, 30 mM) did not affect the rate of relaxation during the initial approximately 50 ms, but accelerated the slower phase of relaxation, consistent with a cyclic cross-bridge model in which Pi increases the proportion of cross-bridges in detached ('weakly bound') states.(ABSTRACT TRUNCATED AT 400 WORDS) Bioorg Khim, 1993 Jan, 19(1), 75 - 80 {Cloning the Staphylococcus aureus enterotoxin B gene, obtained by polymerase chain reaction, and its expression in Escherichia coli cells}; Ignatov KB et al.; To determine the Staphylococcus aureus enterotoxigenicity, we have developed an approach based on polymerase chain reaction (PCR) . Using this method several S . aureus strains have been screened for the presence of the enterotoxin B gene . A DNA fragment of the selected strain (FRI 722H) containing enterotoxin B gene has been obtained by the PCR method and cloned in the pUC19 vector . It is shown that enterotoxin B with the leader peptide forms insoluble complexes in E . coli cells, whereas the mature toxin is present in cytoplasmic fraction in a soluble form . The recombinant toxin made up for 1.7% of the total cellular protein in E . coli JM 109 cells. Eur J Clin Microbiol Infect Dis, 1993, 12 Suppl 1, S13 - 8 Genetic control of population structure in heterogeneous strains of methicillin resistant Staphylococcus aureus; de Lencastre H et al.; Cultures of heterogeneous strains of methicillin resistant Staphylococcus aureus are composed of several subpopulations of cells that differ in their level of antibiotic resistance and frequencies . Experimental evidence is presented indicating that the number of these subpopulations, their resistance level and the frequency with which they are represented in a culture are strain-specific and reproducible with a surprising degree of precision from one culture to another . This implies a genetic control of population structure in these bacteria. Microbiol Immunol, 1993, 37(1), 29 - 34 Efficient adsorption of lysostaphin on bacterial cells of lysostaphin-resistant Staphylococcus aureus mutant; Sakurada J et al.; A simple and efficient method for the purification of staphylolytic endopeptidase (lysostaphin) contained in culture supernatant of Staphylococcus simulans biovar staphylolyticus strain by adsorption of the enzyme on bacterial cells of lysostaphin-resistant S . aureus mutant was successfully devised . Lysostaphin was sufficiently absorbed on the heat-killed mutant cells derived from S . aureus Cowan I and efficiently eluted by 3 M KSCN . Enzyme preparation obtained by a single procedure of the affinity purification was pure enough for practical use . The yield of the enzyme was 25 mg from 1 liter culture and recovery rate was 64%. Monaldi Arch Chest Dis, 1993, 48(1), 29 - 33 Thymomodulin enhances phagocytic and intracellular killing activities of polymorphonuclear leucocytes without increasing release of chemotactic factors; Lantero S et al.; Thymomodulin, a calf thymus derivative, is able to stimulate T-lymphocytes and monocytes, and to activate phagocytes and their precursors . However, it is not fully understood whether the effect of thymomodulin on phagocytic cells is a direct stimulation, or a phenomenon mediated by cytokines released by mononuclear cells . To answer this question, we first evaluated the effects of thymomodulin on the phagocytosis and intracellular killing of Staphylococcus aureus by blood polymorphonuclear cells (PMNs), cultured with or without autologous mononuclear cells . Secondly, during the processes of phagocytosis and intracellular killing, we evaluated the release by PMNs of chemotactic factors for PMNs, lymphocytes and monocytes . No difference was found in the phagocytosis and killing processes when PMNs were incubated alone or with autologous mononuclear cells (p > 0.05, each comparison) . Thymomodulin was able to increase the phagocytosis process when PMNs were incubated with lymphocytes and monocytes (p = 0.05), and to enhance the killing by PMNs cultured alone (p = 0.05), or cultured with autologous mononuclear cells (p < 0.05) . The release of chemotactic factors for PMNs, lymphocytes and monocytes in the supernatants of the phagocytosis experiments, was higher when PMNs were incubated with mononuclear cells, compared to cultures of PMNs alone (p < 0.05, each comparison); and thymomodulin did not increase their release without the presence of autologous mononuclear cells in the cultures (p > 0.05 each comparison) . These data suggest that thymomodulin acts upon PMNs, inducing mononuclear cells to release factors able to stimulate the phagocytosis and the intracellular killing of exogenous organisms, but does not amplify the immune reaction enhancing further leucocytes recruitment. Biofizika, 1993 Jan-Feb, 38(1), 37 - 46 {Molten globule unfolding by strong denaturing agents occurs by the "all or nothing" principle}; Uverskii VN et al.; The equilibrium Gu-HCl-induced denaturation of bovine carbonic anhydrase B and beta-lactamase from Staphylococcus aureus was studied at 4 degrees C by the multiparametric approach . With the use of fast protein size-exclusion chromatography (FPLC) it has been shown that in the region of the molten globule-random coil transition the distribution function of the protein molecules on size is bimodal, i.e . the protein molecules (which are already denatured) can only in one of two conformational states . Consequently the unfolding of the molten globule can be of "all-or-none" character . This means that a protein molecule can be at least in three discrete states: the native, the molten globule and unfolded. Surg Today, 1993, 23(1), 4 - 8 A study on postoperative enteritis caused by methicillin-resistant Staphylococcus aureus; Takesue Y et al.; We investigated the production of staphylococcal enterotoxin (SE) with respect to coagulase types by methicillin-resistant Staphylococcus aureus (MRSA) . A total of 138 strains of MRSA, which were isolated from clinical materials in the surgical ward between 1983 and 1990, were studied . Coagulase type IV strains produced SE A only, whereas coagulase type II strains were classified into four groups by SE production: SE B producing strains (32.7%), SE C producing strains (29.8%), SE B and C coproducing strains (12.5%), and SE A and C coproducing strains (25.0%) . Almost all of the organisms (nine of ten) which were isolated from the feces of patients with MRSA enteritis were SE A and C coproducing strains . The coincidence in time of the prevalence of MRSA enteritis and the isolation SE A and C coproducing strains also demonstrated that these strains caused MRSA enteritis . Although SE C producing strains and SE A and C coproducing strains were simultaneously prevalent in 1990, the former tended to be sensitive while the latter tended to be resistant to minocycline . Considering the variety of antibiotic sensitivity in coagulase type II strains, it is thus considered to be of critical importance for epidemiologic purposes to further characterize isolates by SE typing. Scand J Infect Dis, 1993, 25(1), 123 - 32 Activity of erythromycin and clindamycin in an experimental Staphylococcus aureus infection in normal and granulocytopenic mice . A comparative in vivo and in vitro study; Hoogeterp JJ et al.; The activity of 2 bacteriostatic antibiotics, erythromycin and clindamycin, against Staphylococcus aureus was studied in vitro and in an experimental infection in granulocytopenic as well as normal mice . In vivo, on the basis of dosage, erythromycin was 2.20 times more potent than clindamycin in normal mice and 1.95 times more potent in irradiated granulocytopenic mice . Tissue concentrations were calculated from the plasma concentrations, taking plasma protein and tissue binding into account . On the basis of the area under the curve for the free tissue concentrations, clindamycin was 1.48 times more potent than erythromycin in normal mice and 1.64 times more potent in irradiated mice, which is somewhat less than expected from the comparison in vitro . It is concluded that the relative antistaphylococcal efficacies of erythromycin and clindamycin in vivo can be predicted from the in-vitro values if the relevant pharmacokinetics are taken into account . Irradiation decreased the efficacy of both antibiotics such that an about 8-fold increase in dose led to an antibacterial effect similar to that in non-irradiated animals . This could imply that these drugs will not be sufficiently effective against staphylococcal infections in granulocytopenic patients. Scand J Infect Dis, 1993, 25(1), 107 - 13 The value of surveillance cultures in neutropenic patients receiving selective intestinal decontamination; de Jong PJ et al.; 230 neutropenic episodes in 84 patients with acute myeloid leukemia receiving selective intestinal decontamination were studied to evaluate the ability of surveillance cultures to monitor the efficacy of microbial suppression, to identify causative organisms in case of fever, and to predict infection due to potential pathogens (i.e . Staphylococcus aureus and aerobic Gram-negative bacteria) . Most cultures became negative soon after the administration of prophylactic antibiotics and there were only few persistent colonizations . 14 potential pathogens resistant to the intestinal decontamination regimen were isolated in surveillance cultures, none of which caused infection . Of the 212 febrile episodes, only 22 were caused by a microbiologically documented infection with potential pathogens . Most microbiologically documented infections were caused by organisms not routinely identified by surveillance cultures, indicating efficient selective intestinal decontamination . Only 9 (41%) of the 22 infections with potential pathogens were predicted by surveillance cultures . We conclude that surveillance cultures are of limited use in predicting infection or identifying causative organisms of fever in neutropenic patients receiving selective intestinal decontamination . However, they are useful in monitoring the efficacy of microbial suppression . One set of surveillance cultures each week after the disappearance of potential pathogens would be sufficient. Nephron, 1993, 63(4), 409 - 15 Depressed phagocytosis in hemodialyzed patients: in vivo and in vitro mechanisms; Vanholder R et al.; Infection is a frequent complication and the major cause of death among end-stage renal patients . Polymorphonuclear phagocytes (PMNL) are important in host defense mainly because of bacterial destruction by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-related free radical production following phagocytosis . In this study, hexose monophosphate pathway glycolytic activity, delivering energy to NADPH oxidase, is evaluated in vivo and in vitro, in healthy controls and in dialyzed renal failure patients . Our results show a marked parallel and correlated inhibition in the response to three stimuli for phagocytic activity (Staphylococcus aureus, formyl-methionine-leucine-phenylalanine, phorbol myristic acid) in predialysis samples . These data point to a main suppression of metabolic pathways, possibly beyond protein kinase C . This response is further suppressed at the 15th minute of cuprophane dialysis, for all stimuli studied (-40 to -94%; p < 0.001) except PMA . PMNL response remains intact during dialysis with non-complement-activating dialyzers . In vitro experiments confirm decreased PMNL glycolytic activity after the suspension of cuprophane fragments in normal whole blood . We conclude that polymorphonuclear cell energy delivery to NADPH oxidase is impaired in patients with end-stage renal failure . The impaired response against various stimuli is different in predialysis blood samples compared to samples collected during cuprophane dialysis, and may be related to two different conditions . These events probably contribute to the acquired immune suppression of uremia and the high incidence of infection among dialysis patients. Jpn J Antibiot, 1993 Jan, 46(1), 8 - 17 {Effects of a combination therapy consisting of arbekacin and ampicillin of methicillin-resistant Staphylococcus aureus}; Deguchi K et al.; Investigations were made on effects of a combination therapy consisting of arbekacin (ABK) and ampicillin (ABPC) against methicillin-resistant Staphylococcus aureus (MRSA) . The following conclusions were drawn . 1 . A frequency of FIC index to be < or = 0.5 upon a combination therapy consisting of ABK and ABPC against MRSA was 48.7%, hence this therapy showed better antibacterial effects than the other 8 combination therapies consisting of ABK and other antibiotics previously studied . 2 . A combination therapy consisting of ABK and ABPC against MRSA was better than any other combination therapy because ABPC had a higher binding affinity with penicillin-binding protein 3 of MRSA, and because a high antibacterial activity of ABPC was maintained during the combination therapy . ABPC did not lose antibacterial activity largely because MRSA dose not produce beta-lactamase, or most of MRSA produces little beta-lactamase activity . 3 . Our data indicate that the success of a combination therapy against MRSA depends upon the effects of the drug under sub MIC concentrations. J Burn Care Rehabil, 1993 Jan-Feb, 14(1), 55 - 7 Development of septic arthritis by hematogenous seeding in a pediatric patient with burns; Smoot EC et al.; We describe a pediatric patient with burns who experienced a septic hip joint from hematogenous seeding by sustained Staphylococcus aureus bacteremia . A search for a central vascular focus of the bacteremia included the use of an indium scan, which localized purulence within the left hip joint . The physical findings, radiographic findings, and subsequent aspiration of the hip joint were diagnostic for a septic arthritis . This case alerts the physician that the pediatric hip is a potential site for infection in the presence of bacteremia . Once infected, the joint may become a secondary source of an ongoing bacteremia . The diagnostic work-up and treatment for a septic hip have been discussed . This occult site of infection is easy to overlook because a painful hip joint in the pediatric patient with burns may be confused for painful burn wounds . The prerequisites to joint infections in this population were all present: bacteremia, intercurrent illness, unique vascular anatomy, and an impaired immune response. Acta Paediatr, 1993 Jan, 82(1), 113 - 5 Pyomyositis and staphylococcal scalded skin syndrome; Wong GW et al.; Pyomyositis is a rare musculoskeletal infection in non-tropical countries . We report a child who had pyomyositis complicated by staphylococcal scalded skin syndrome . This complication has not previously been described in patients with pyomyositis . Early diagnosis of pyomyositis was made by ultrasound examination, and percutaneous needle drainage under ultrasound guidance was performed . Pus aspirated and blood cultures grew Staphylococcus aureus . No open surgical drainage was required and resolution of the abscess was documented by serial ultrasound examinations. Kansenshogaku Zasshi, 1993 Jan, 67(1), 45 - 52 {Hospital infection with methicillin-resistant Staphylococcus aureus (MRSA) in Saga Medical School Hospital, a rapid increase in coagulase type-VIII strains}; Nagasawa Z et al.; Hospital infection with MRSA has increased in Saga Medical School Hospital . The causative MRSA consisted predominantly of coagulase type-II strain before 1989, but after 1990, coagulase type-VII MRSA increased rapidly . This type-VII strain has marked multiple drug-resistance, and the pattern of drug sensitivity of MRSA in this hospital was different from that of MRSA detected in other facilities, which are clinically serious problems, therefore, we conducted an etiological study of the background of the increase in MRSA infection in our hospital . The results of the study are summarized as follows: 1) The proportions of MRSA (on strain from one patient) to all types of S . aureus detected in the hospital were 26% for 1986, 23% for 1988, 37% for 1989, 30% for 1990 and 60% for 1991 . The proportion increased greatly in 1991 . 2) Coagulase type VII-MRSA was first detected only in 5 patients in 1989, then it tended to spread, and this type (probably derived from the same strain) accounted for 47% of MRSA infection in patients examined in 1991 . 3) The study of the drug sensitivity pattern and etiological survey of the infection showed that coagulase type VII-MRSA prevalent in the hospital consisted of two types: CLDM, and EM-sensitive, IPM/CS, and MINO-resistant and TSST-1 non-producing and enterotoxin non-producing type, and CLDM, and EM-sensitive, IPM/CS, and MINO-resistant and TSST-1 non-producing type with enterotoxin serotype A . 4) Coagulase type VII-MRSA (Probably derived from the same strain) was detected in physicians and nurses working in affected wards and also in the patients's room.(ABSTRACT TRUNCATED AT 250 WORDS) Clin Immunol Immunopathol, 1993 Jan, 66(1), 43 - 51 Regulation of B cell function by bucillamine, a novel disease-modifying antirheumatic drug; Hirohata S et al.; Bucillamine {N-(2-mercapto-2-methylpropionyl)-L-cysteine} (BUC) is a thiol compound that differs from D-penicillamine (DPC) in that it contains two free sulfhydryl groups . Clinical trials have demonstrated that its efficacy in rheumatoid arthritis is superior to that of DPC, but its mechanism of action remains unclear . We therefore examined the effects of BUC on the in vitro function of human B cells in comparison to those of DPC . IgM production was induced from highly purified B cells from healthy donors by stimulation with Staphylococcus aureus Cowan I (SA) plus factors generated from mitogen-activated T cells (TF) or interleukin-2 (IL-2) or with immobilized anti-CD3-activated CD4+ T cells . BUC suppressed the production of IgM at concentrations of 0.3-100 micrograms/ml irrespective of the presence of CuSO4 . Whereas BUC suppressed the production of IL-2 and interferon-gamma by immobilized anti-CD3-activated CD4+ T cells, its suppressive effects on the production of IgM in anti-CD3-stimulated cultures were not overcome by addition of TF or IL-2, indicating that the action of BUC involves direct inhibition of B cell function . BUC suppressed the initial stages of B cell activation, but not the maturation of previously activated B cells . In contrast to DPC, the suppressive activities of BUC did not require the presence of copper and were not overcome by the addition of monocytes or catalase . The effects of SA981, a metabolite of BUC with an intramolecular disulfide, on B cell function were more marked than those of BUC, whereas the effects of SA679, another metabolite of BUC with one of the two sulfhydryl bonds methylated, were similar to those of DPC . SA672, a metabolite of BUC with both of the two sulfhydryl bonds methylated, did not suppress B cell function . These results indicate that BUC as well as some of its metabolites inhibit cytokine production by T cells and also suppress the production of IgM at least in part by directly inhibiting B cells . These compounds exert immunosuppressive effects that are similar to those of DPC, but also unique inhibitory effects that depend upon the capacity of BUC to form an intramolecular disulfide between its two sulfhydryl groups. Mol Gen Genet, 1993 Jan, 236(2-3), 187 - 92 LamB as a carrier molecule for the functional exposition of IgG-binding domains of the Staphylococcus aureus protein A at the surface of Escherichia coli K12; Steidler L et al.; One, two or four IgG-binding domains of the Staphylococcus aureus Protein A (SPA) were inserted into the LamB protein which was expressed under control of the tac promoter . The chimeric proteins were shown to be exposed at the cell surface by analysis of isolated outer membranes and also by testing their functional interaction with IgG molecules . We hereby show that the LamB protein can accept as many as 232 amino acids (four SPA domains) and still be incorporated into the Escherichia coli outer membrane, while maintaining the functional conformation of the inserted SPA polypeptides. Int J Pept Protein Res, 1993 Jan, 41(1), 74 - 84 Sequence and solution conformation of the 20-residue peptaibols, saturnisporins SA II and SA IV; Rebuffat S et al.; Saturnisporins SA II and SA IV are the major components of the 20-residue peptaibol mixture isolated from a culture of the fungus Trichoderma saturnisporum . These peptides exhibit antibiotic activity against Staphylococcus aureus . Their sequences were derived from unequivocal methodology implying the combined use of positive ion FAB mass spectrometry and NMR: the majority of the sequences result from mass spectrometry fragmentations and the location of isomeric residues arises either from analysis of ROESY cross-peaks between contiguous amide protons or from heteronuclear 2J or 3J 1H-13C couplings detected in long-range 1H-13C COSY experiments . The sequence specific 1H and 13C NMR assignments are described . Saturnisporins SA II and SA IV exhibit similar secondary structures, as deduced from their ROESY patterns and 3JNHC alpha H coupling constant values, together with amide hydrogen-deuterium exchange rates and temperature coefficients of amide and carbonyl groups . An overall alpha-helical structure is proposed, maintaining two regions of distortion to this regular structure; i) the N-terminal part, which contains 3(10) and mixed alpha-3(10) turns, and ii) the Aib10-Val15 region, including a Pro residue which accommodates a bend stabilized by two 3(10) H-bonds. Antimicrob Agents Chemother, 1993 Jan, 37(1), 32 - 8 High-level mupirocin resistance in Staphylococcus aureus: evidence for two distinct isoleucyl-tRNA synthetases; Gilbart J et al.; Mupirocin resistance in Staphylococcus aureus results from changes in the target enzyme, isoleucyl-tRNA synthetase (IRS) . Twelve strains of S . aureus comprising four susceptible (MICs < or = 4 micrograms/ml), four intermediate level-resistant (MICs between 8 and 256 micrograms/ml), and four highly resistant (MICs > or = 512 micrograms/ml) isolates were examined for their IRS content and the presence of a gene known to encode high-level mupirocin resistance . Ion-exchange chromatography of cell extracts showed a single IRS active peak in mupirocin-susceptible strains, with 50% inhibitory concentrations (IC50s) of 0.7 to 3.0 ng of mupirocin per ml . In strains showing intermediate mupirocin resistance, similar single IRS activity peaks were observed, but these were less sensitive to inhibition, and the mupirocin IC50s for them were 19 to 43 ng/ml . Strains that were highly resistant to mupirocin displayed two distinct peaks; one was similar to that found with susceptible strains (IC50, 0.9 to 2.5 ng/ml), but an additional peak with an IC50 of 7,000 to 10,000 ng/ml was also observed . A strain cured of the plasmid encoding high-level mupirocin resistance lacked the resistant IRS peak . Restriction digests, produced by endonuclease NcoI, of total bacterial DNA isolated from the highly resistant strains hybridized with a mupirocin resistance gene probe, whereas DNA isolated from the intermediate level-resistant and susceptible strains did not . These results demonstrate that two different IRS enzymes were present in highly mupirocin-resistant S . aureus strains . In strains expressing intermediate levels of resistance, only a chromosomally encoded IRS which was inhibited less by mupirocin than IRS from fully susceptible strains was detected. J Steroid Biochem Mol Biol, 1993 Jan, 44(1), 1 - 11 Expression and functional analysis of steroid receptor fragments secreted from Staphylococcus aureus; Cao X et al.; Fragments of the DNA-binding domain of the rat glucocorticoid receptor (rGR) and the human estrogen receptor (hER) were expressed in Staphylococcus aureus as a chimeric fusion to protein A by using a modified expression vector with an artificial factor X-cleavage site . The secreted product was isolated by hydrophobic chromatography on Phenyl-Sepharose and purified on DNA-cellulose or by anion-exchange chromatography . After cleavage of the protein A moiety, the purified rGR DNA-binding domain from amino acids 406 to 523 (rGR406-523), binds specifically to a glucocorticoid responsive element as a homodimer but cannot form heterodimers with the DNA-binding domain of the hER . Amino acids 510 to 523 following the zinc finger region, as well as free sulfhydryl-groups are necessary for DNA-binding, which is more efficient when the tripeptide Gly-Gly-Cys is added to the carboxy terminal end . Despite its specific interaction with DNA, rGR406-523 does not activate transcription from the MMTV promoter in a cell-free system that efficiently responds to addition of native GR, suggesting that regions essential for transcriptional activation in vitro are located outside of the DNA-binding domain. Eur J Immunol, 1993 Jan, 23(1), 191 - 9 Regulation of T helper cell cytokine expression: functional dichotomy of antigen-presenting cells; Schmitz J et al.; A bias to either cell-mediated or antibody-mediated effector mechanisms is induced in an immune response against a pathogen, if activated T helper cells (Th) predominantly express Th1 {interleukin (IL)-2, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-beta} or Th2 (IL-4, IL-5, IL-6 and IL-10) cytokines . Here we provide evidence that, due to the capability to secrete IL-1, macrophages, but not B cells, as antigen-presenting cells (APC) induce production of IFN-gamma in resting Th cells . Normal murine splenic Th cells were activated in vitro with the superantigen Staphylococcus aureus enterotoxin B (SEB) presented by macrophages as compared to other APC from murine spleen . As determined by immunofluorescence, Th cells producing IL-2 but almost none producing IL-4 and IL-5 are generated, irrespective of the type of APC . Generation of IFN-gamma-producing Th cells is largely dependent on presentation of SEB by macrophages . The requirement for macrophages, however, is overcome if IL-1 is provided . Expression of IFN-gamma by Th cells is not induced, if production of IL-1 by macrophages is inhibited by IL-10 . Our results suggest a functional dichotomy of APC: normal resting Th cells differentiate into IL-2 and IFN-gamma secreting cells (Th1 cells) if antigen is presented by macrophages, whereas presentation by B cells generates Th cells secreting IL-2, which might differentiate into Th2 cells upon re-stimulation. Cutis, 1993 Jan, 51(1), 43 - 4 Linear bacterial dissection; Shelton R; An unusual case of an erythematous indurated arcuate plaque with a curvilinear appearance was noted after a patient manipulated an ingrown beard hair . During removal of a biopsy specimen, a purulent discharge was expressed that had evidently dissected through the dermis forming this interesting shape . This may be similar to a recently reported entity described by Shelley and Shelley as linear bacterial dissection . Culture of the discharge grew Staphylococcus aureus and Klebsiella pneumoniae . Complete resolution occurred after drainage and antibiotic therapy . Cellulitis and erysipelas typically do not show central clearing or form arcuate plaques . Aside from lymphangitis, thrombophlebitis, and sinus tracts of carbuncles, dermal infectious processes rarely present as linear or arcuate plaques . The differential diagnosis and a review of the literature are presented. Intensive Care Med, 1993, 19(5), 285 - 9 Resistance to methicillin and virulence of Staphylococcus aureus strains in bacteriemic cancer patients; Marty L et al.; OBJECTIVE: To estimate the relation between resistance to methicillin and virulence in strains of Staph . aureus by estimating the survival at day 30 . DESIGN: Case control analysis . SETTING: Inpatients at a major Cancer Referral Center . PATIENTS: 21 patients with methicillin-resistant Staph . aureus (MRSA) bactaeremia and 45 patients with methicillin-susceptible Staph . aureus (MSSA) bactaeremia, all treated with vancomycin . MEASUREMENTS AND MAIN RESULTS: Fewer MRSA than MSSA patients were neutropenic (p = 0.04), but more MRSA patients were hospitalized in intensive care units (p = 0.03), had had surgical episodes (p = 0.06) . MRSA patients also had more microbiological samples cultured (p = 0.007) . The outcome of the bacteriemic episodes in 14 MRSA and 14 MSSA patients matched for these four factors showed that blood cultures from MRSA patients remained positive significantly longer (p = 0.04), but that survival and length of hospital stay were not significantly different . CONCLUSIONS: These results do not indicate that methicillin-resistance in Staph . aureus is associated with increased virulence. Perit Dial Int, 1993, 13 Suppl 2, S79 - 80 Establishing an experimental infection model for peritoneal dialysis: effect of inoculum and volume; Calame W et al.; The effect of the number of bacteria and the volume of the inoculum was studied in an experimental infection model to establish a peritoneal dialysis model in the rat . Staphylococcus aureus was used in all experiments, and Staphylococcus epidermidis only in the volume experiments . A bacterial number between 10(8) and 10(9) colony-forming units (cfu) resulted in a time-dependent decrease of bacteria collected from the peritoneal cavity . Higher concentrations resulted in the death of animals, while lower concentrations were rapidly cleared . There was a positive correlation between the volume in which 3 x 10(8) cfu were dissolved and the number of bacteria isolated from the peritoneal cavity 24 hours after infection . The results of this study led to an experimental dialysis model using 10 mL of dialysis fluid and 0.5 mL of a suspension containing 3 x 10(8) cfu of Staphylococcus aureus. Perit Dial Int, 1993, 13 Suppl 2, S332 - 4 Staphylococcus aureus peritonitis is associated with Staphylococcus aureus nasal carriage in peritoneal dialysis patients; Piraino B et al.; Although the definition of S . aureus nasal carriage in peritoneal dialysis patients is variable, carriage is often defined as 2 or more positive cultures for S . aureus . We investigated the relationship between S . aureus infections and nasal carriage (defined as 1 or more positive nose cultures) in 138 patients on peritoneal dialysis . By this definition, approximately 50% of the patients were carriers . The rates of S . aureus exit-site infection (0.28/year vs 0.43/year, p < 0.001) and peritonitis (0.08/year vs 0.20/year, p < 0.001) were lower in the noncarriers (patients with no positive nose cultures) than in those patients who had 1 or more positive nose cultures . Patients with only 1 positive nose culture had a similar S . aureus exit-site infection rate (0.31/year vs 0.28/year), but a higher peritonitis rate (0.24/year vs 0.08/year, p < 0.001) compared to those patients with no positive nose cultures . However, patients with 2 or more positive nose cultures were at increased risk for both S . aureus exit-site infections and peritonitis . This relationship held even when only patients with frequent nose cultures were included in the analysis . Thus patients with even 1 positive nose culture for S . aureus are at risk for S . aureus peritonitis and should not be classified as noncarriers. Exp Clin Immunogenet, 1993, 10(1), 16 - 20 Defective calcium-dependent signal transduction in B lymphocytes of a certain common variable immunodeficiency; Kondo N et al.; Two different common variable immunodeficiency patients were studied . Patient 1 showed hypogammaglobulinemia in all major classes of gammaglobulins and slightly reduced percentages of circulating CD19+ cells, CD20+ cells and surface IgG-or IgA-bearing cells with a normal percentage of surface IgM-bearing cells . Patient 2 showed normal levels in IgM with IgG and IgA deficiencies and almost normal percentages of circulating CD19+ cells, CD20+ cells and surface IgM-, IgG- or IgA-bearing cells . The proliferative responses of peripheral blood mononuclear cells to B cell mitogen, Staphylococcus aureus Cowan I, were reduced in patient 1 but not in patient 2 . The intracellular calcium concentrations in lymphoblastoid cell lines (LCLs) transformed by Epstein-Barr virus from patient 2 were increased after anti-mu stimulation . The concentrations were scarcely increased in LCLs from patient 1 . These results suggest that B cells of patient 1 have a defective Ca(2+)-dependent signal transduction pathway, resulting in a failure of cell activation and cell proliferation . The defect in B cells of patient 2 may exist on immunoglobulin gene switching or expression rather than the Ca(2+)-dependent signal transduction pathway. Nephrol Dial Transplant, 1993, 8(3), 235 - 9 Nasal mupirocin ointment decreases the incidence of Staphylococcus aureus bacteraemias in haemodialysis patients; Boelaert JR et al.; The incidence of S . aureus bacteraemia in a haemodialysis unit was studied over 2 years (167.75 patient-years of follow-up) during which nasal calcium mupirocin was used to eradicate nasal S . aureus carriage; this incidence was compared to that previously observed in the same unit before the use of nasal mupirocin (185.8 patient-years) . Nasal mupirocin led to eradication of nasal S . aureus carriage in 96.3% of surveillance cultures and to a fourfold reduction in the incidence of S . aureus bacteraemia per patient-year, from 0.097 before mupirocin to 0.024 with mupirocin use (P = 0.008) . Once or thrice weekly maintenance regimens of mupirocin were equally efficacious . The incidence of bacteraemia caused by other micro-organisms was not significantly affected . One single mupirocin-resistant isolate was identified in a nasal surveillance culture . Eradication of S . aureus from the nares did not lead to overgrowth by other micro-organisms . Chemoprophylaxis with nasal mupirocin in haemodialysis patients is cost-effective. Nephrol Dial Transplant, 1993, 8(2), 157 - 62 Adherence of Staphylococcus aureus to cultures of human peritoneal mesothelial cells; Glancey G et al.; Strains of Staphylococcus aureus, isolated from the effluent of patients with peritonitis on CAPD (continuous ambulatory peritoneal dialysis), adhered well to both cultured human mesothelial cells and to fibronectin, but not to laminin or gelatin . Mesothelial cells grown in medium M199 exhibited more surface fibronectin compared to cells grown in MEM-Dval and demonstrated higher levels of S . aureus adherence . Soluble fibronectin concentrations up to 10 micrograms/ml increased the adherence of S . aureus to cultured mesothelial cells . The dose-response curve was consistent with the binding of fibronectin to a saturable receptor of apparent dissociation constant (KD) = 1.7 x 10(-10) M . This corresponds closely to the KD (2 x 10(-10) M) of the staphylococcal fibronectin-binding protein . S . aureus adherence was increased following the preincubation of mesothelial cell monolayers with interleukin-1 and was maximal after 6 h preincubation . Treating mesothelial cells with interferon-gamma for 48-72 h reduced the adherence of S . aureus. Mol Microbiol, 1993 Jan, 7(1), 99 - 107 A collagen receptor on Staphylococcus aureus strains isolated from patients with septic arthritis mediates adhesion to cartilage; Switalski LM et al.; Staphylococcus aureus strains isolated from patients with septic arthritis or osteomyelitis possess a collagen receptor present in two forms, which contains either two or three copies of a 187-amino-acid repeat motif . Collagen receptor-positive strains adhered to both collagen substrata and cartilage in a time-dependent process . Collagen receptor-specific antibodies blocked bacterial adherence, as did preincubation of the substrate with a recombinant form of the receptor protein . Furthermore, polystyrene beads coated with the collagen receptor bound collagen and attached to cartilage . Taken together, these results suggest that the collagen receptor is both necessary and sufficient to mediate bacterial adherence to cartilage in a process that constitutes an important part of the pathogenic mechanism in septic arthritis. Infect Control Hosp Epidemiol, 1993 Jan, 14(1), 29 - 35 Restriction endonuclease analysis of plasmid DNA from methicillin-resistant Staphylococcus aureus: clinical application over a three-year period; Trilla A et al.; OBJECTIVE: To investigate trends in plasmid types of methicillin-resistant Staphylococcus aureus over a 3-year period and determine the clinical impact of plasmid typing . DESIGN: Restriction endonuclease analysis of plasmid DNA (REAP) was used to type 270 clinical isolates . SETTING: A Veterans Affairs Medical Center . RESULTS: Forty-four different REAP types were identified and, of these, 23 appeared only once . Types designed as V1 and V2 were more commonly found in nosocomial cases than in community-acquired cases (p < .05); whereas the K and D types were more common in isolates from community-acquired cases (p < .05) . In 30 (11%) isolates, no plasmid was detected . When combined with epidemiological data, REAP typing revealed 4 small outbreaks that would have been missed using traditional epidemiological methods alone . In large outbreaks involving 10 or more cases, REAP typing data revealed unsuspected patterns of transmission . CONCLUSIONS: Multiple plasmid types were present in this endemic setting . Restriction endonuclease analysis of plasmid DNA was a practical and valuable adjunct to traditional epidemiological methods. Diagn Microbiol Infect Dis, 1993 Jan, 16(1), 9 - 16 Molecular epidemiology and antimicrobial susceptibility testing . Testing of quinolone-resistant Staphylococcus aureus strains isolated in Brazil; Rodrigues JN et al.; An outbreak of 20 oxacillin-resistant Staphylococcus aureus (ORSA) bloodstream infections (BSIs) was detected in the intensive care unit (ICU) at Hospital Sao Paulo, Brazil . In a surveillance study, 10% of ICU personnel were defined as chronic nasal carriers of ORSA . Thirteen BSIs and five nasal isolates were available for phage typing, restriction endonuclease analysis of plasmid (REAP) and chromosomal DNA hybridized with 32P rRNA gene probe . Susceptibility testing against select antimicrobial agents, including 11 quinolones, six glycopeptides, and five topical agents, was performed by broth microdilution and the disk diffusion tests . Ten of the 13 BSI isolates and four of the five nasal strains were oxacillin and quinolone resistant . The new fluoroquinolones CI-960 and WIN57273, the glycopeptides, and the topical agents ramoplanin, bacitracin, mupirocin, and novobiocin were most active . Fourteen strains, 12 BSI isolates, and three nasal isolates showed the same REAP profile . Moreover, the same REAP and chromosomal profile was detected in at least nine BSI isolates and in two nasal isolates . These strains were nontypable by phage typing . We concluded that nosocomial cross-transmission of a single, multiresistant strain of S . aureus occurred and that the epidemic reservoir was nasal carriage by ICU personnel. Cell Immunol, 1993 Jan, 146(1), 117 - 30 Intracellular signaling events associated with the induction of DNA synthesis in human B lymphocytes . I . Stimulation of PKC-dependent and -independent pathways by LMW-BCGF; Kolb JP et al.; The low molecular weight B cell growth factor (LMW-BCGF) induces the G1 --> S transition in human B lymphocytes activated by a first signal, Staphylococcus aureus Cowan (SAC) or anti-mu antibody . It also stimulates proliferation of normal long-term B cell lines and some B cell tumors . We have previously reported that LMW-BCGF induces the hydrolysis of polyphosphoinositides (PI) and a rise in intracellular free calcium concentration, through the generation of inositol trisphosphate (InsP3) (Renard et al., J . Immunol . 18, 1705, 1988) . In the present work we have analyzed the possible association between early signaling events elicited by LMW-BCGF in SAC-activated B cells and its ability to provoke DNA synthesis, notably at the level of phospholipase C (PLC) and protein kinase C (PK-C) activation . Inhibitors of PLC and of InsP3-induced calcium release were found to block LMW-BCGF-dependent DNA synthesis . An increase in membrane-associated protein kinase C (PK-C) activity was detected after the addition of the growth factor and the mitogenic effect of LMW-BCGF was partially suppressed when B cell blasts were incubated with staurosporine or H-3, two inhibitors of PK-C activity . In addition, the mitogenic effect due to the addition of LMW-BCGF was not modified by the incubation of B cell blasts with high concentrations of TPA, even if this treatment inhibited cellular response to a low concentration of TPA . LMW-BCGF also increased intracellular pH in B cell blasts and lymphokine-induced mitogenic activity was reduced when the Na+/H+ amiloride or ethylisopropyl amiloride (EIPA) antiport blockers were added . These results suggest that (i) LMW-BCGF-induced PI breakdown and CA2+ mobilization and cell alkalinization are associated with the induction of cell proliferation, and (ii) the activation of PK-C does not appear to be the sole pathway activated by LMW-BCGF. Biochem J, 1993 Jan 1, 289 ( Pt 1), 289 - 97 Deglycosylation and fragmentation of purified rat liver angiotensin II receptor: application to the mapping of hormone-binding domains; Desarnaud F et al.; We report new structural data about the rat liver angiotensin II receptor, which belongs to the AT1 subclass . This receptor has been purified at analytical or semi-preparative levels by a previously described strategy involving its photolabelling with a biotinylated azido probe and selective adsorption of the covalent probe-receptor complexes to immobilized streptavidin {Marie, Seyer, Lombard, Desarnaud, Aumelas, Jard and Bonnafous (1990) Biochemistry 29, 8943-8950} . Chemical or enzymic deglycosylation of the purified receptor has shown a shift in its molecular mass from 65 kDa to 40 kDa . Fragmentation of the purified receptor was carried out with V8 protease from Staphylococcus aureus, CNBr and trypsin . It was possible to find trypsin-treatment conditions which allowed production of a 6 kDa probe-fragment complex with a satisfactory yield . Attempts to localize this small fragment (5 kDa after subtraction of the probe contribution) in the recently published rat AT1 receptor sequence are reported . As expected, this fragment is not glycosylated; moreover, its further fragmentation by CNBr induces a very slight decrease in its size . These data support the hypothesis that a receptor sequence comprising the third transmembrane domain and adjacent portions of extra- and intracellular loops is involved in photolabelling by the C-terminal azidophenylalanine of the angiotensin-derived probe . These preliminary results are discussed in terms of future prospects for the characterization of hormone-binding domains of angiotensin II receptors. Vasa, 1993, 22(3), 241 - 6 {Pathogenesis and size development of infected saccular aneurysms--an animal experiment study}; Bosse U et al.; Both femoral arteries of 56 Wistar rats were intersected and re-anastomosed by microsurgical technique . 24 anastomoses were performed under aseptic conditions and 88 were contaminated with a standard suspension of staphylococcus aureus . All the contaminated anastomoses which had remained patent developed true saccular aneurysms that reached their maximum volume during the acute period of infection between the 10th and 50th postoperative day . None of the anastomoses performed under aseptic conditions developed an aneurysm . This experiment represents an animal model for the reproduction of true saccular aneurysms of the arterial wall . It may help to investigate the etiology of arterial aneurysm formation. Microbios, 1993, 75(302), 17 - 21 Survival ability of clinical isolates of methicillin-resistant Staphylococcus aureus; Sasatsu M et al.; The survival abilities of methicillin-resistant strains of Staphylococcus aureus were compared with those of sensitive strains . Absorbent cotton was inoculated with 10(8) cells of each strain tested and stored at 30% humidity at 25 degrees C . The numbers of cells of sensitive strains decreased significantly to well below those of resistant strains after 4 weeks, while cells of resistant strains remained above 10(3) after 14 weeks. Cytobios, 1993, 75(300), 21 - 32 Morphological changes of Carcinoscorpius rotundicauda amoebocyte and Escherichia coli during their interaction; Ding JL et al.; Interaction between Carcinoscorpius rotundicauda amoebocytes and Escherichia coli or Staphylococcus aureus has shown that whilst E . coli was observed to lyse, S . aureus remained intact . The heavily granulated amoebocytes were activated by the lipopolysaccharide of E . coli to undergo dramatic morphological changes with time, leading to degranulation which caused lysis of the E . coli . Degranulation of the amoebocytes involved maturation of the larger electron dense granules (0.7-2.0 micron) which were apparently mobilised towards the cell membrane whilst undergoing a loss in electron density . Some of these larger granules were expelled at 15-30 min while other intracellular granules underwent further degranulation, where parallel filaments appearing like microtubules were evident, giving the granules a striated and less dense form . By 60 min, there was complete depolymerisation of the microtubular filaments giving rise to undefined, fused and homogeneously packed striations and particulate material in the cytoplasm; the smaller electron dense granules (0.3-0.7 micron) remained unchanged . This asynchronous degranulation and preferential expulsion of granules from only some amoebocytes while maintaining others intact, implies that the horseshoe crab has safeguards in its defence mechanism, hence ensuring its continued survival as a 'living fossil'. Biotechnol Ther, 1993, 4(1-2), 31 - 42 Functional interaction between interleukin-9/P40 and interleukin-4 in the induction of IgE production by normal human B lymphocytes; Dugas B et al.; Interleukin-4 (IL-4) has been shown to induce a high rate of IgE production by normal B lymphocytes from peripheral blood without preactivation of these cells . In the present study, significant IgE production was observed at the concentration of 30 U/mL IL-4 (0.5 ng IgE/mL), and a plateau value was reached at 300 U/mL (1.2 ng IgE/mL) . In these experiments, recombinant human (rh) and murine (rm) interleukin-9/P40 (IL-9/P40) were unable to induce such an IgE production by human B lymphocytes . However, in the presence of a suboptimal dose of IL-4 (100 U/mL), both rh and rm IL-9/P40 enhanced the IgE production in a dose-dependent manner (from 3 to 1000 U/mL) . Under these experimental conditions, the slight but significant (p < 0.01, Student's t test) production of IgG (50 ng/mL) was also potentiated when the cell cultures were performed in the presence of 300 U/mL of IL-9/P40 . The IL-4-induced IgG and IgM production by purified B lymphocytes preactivated with Staphylococcus aureus Cowan strain I was also potentiated in the presence of 300 U/mL of rh IL-9/P40, indicating a direct effect of this cytokine on this cell type . In contrast, IgE production by Staphylococcus aureus Cowan strain I-activated B lymphocytes was never observed . Taken together these data suggested a direct role for IL-9/P40 in the regulation of immunoglobulin production by B lymphocytes. Perit Dial Int, 1993, 13(3), 224 - 7 Survival of 138 surgically placed straight double-cuff Tenckhoff catheters in patients on continuous ambulatory peritoneal dialysis; Weber J et al.; OBJECTIVE: To determine the natural history of a surgically placed Tenckhoff catheter in patients on continuous ambulatory peritoneal dialysis (CAPD) . DESIGN: Prospective 7-year study analyzing catheter survival of all catheters using the Kaplan-Meier life table methodology . SETTING: Teaching hospital, department of nephrology . PATIENTS: One hundred and fifteen unselected patients beginning CAPD . INTERVENTIONS: Removal of the catheter required for the following complications: exit-site or tunnel infections or relapsing peritonitis, outflow obstruction, pericatheter leak, and development of hernias . MAIN OUTCOME MEASURES: Period between insertion and removal of the catheter . RESULTS: The cumulative survival of all catheters after 1, 2, and 3 years of CAPD was 87%, 69% and 65% . Catheter survival of the first versus the second catheter after 1 year was significantly longer (p = 0.03) . The difference was not significant in relation to diabetes, age, and sex . Infectious complications caused 61% (n = 19) of all 31 catheter failures, mainly due to tunnel infections caused by Staphylococcus aureus (n = 12) . "Mechanical" complications accounted for 49% (n = 12) of catheter failures . Eight of 12 mechanical complications were outflow failures . Seven patients had to be transferred to hemodialysis . CONCLUSIONS: The straight Tenckhoff catheter is a reliable peritoneal access device for CAPD in an unselected patient population. Perit Dial Int, 1993, 13(3), 198 - 200 Chronic tunnel infections in children: removal and replacement of the continuous ambulatory peritoneal dialysis catheter in a single operation; Schroder CH et al.; OBJECTIVE: Chronic tunnel infections often necessitate the removal of the continuous ambulatory peritoneal dialysis (CAPD) catheter . Most published studies advocate postponing the insertion of a new catheter for several weeks . For young children it will be particularly difficult to wait this length of time, since vascular access may be cumbersome, and hemodialysis may not be well tolerated . The present study describes the results of the simultaneous removal and replacement of the CAPD catheter . DESIGN: Twenty-three Toronto Western Hospital II catheters were inserted in 17 children because of infectious complications (21 chronic tunnel infections; 2 recurrent peritonitis) in a single operation under appropriate antibiotic prophylaxis . The new catheter was inserted at the contralateral side of the abdomen with the deep cuff in the midline, using the same entrance to the peritoneal cavity . Dialysis was resumed immediately after the operation . SETTING: A university pediatric dialysis unit . PATIENTS: Seventeen children (mean age 3.7 years; range 1.0-8.5 years) were studied . In this group 23 catheters were replaced . RESULTS: In four cases a relapse of the tunnel infection was observed within 3 months . All other cases remained free of infection for a period of at least 6 months . The main causative microorganism was Staphylococcus aureus (15 occurrences) . CONCLUSION: It is not necessary to interrupt peritoneal dialysis for the replacement of a CAPD catheter because of infectious complications. Eur J Haematol Suppl, 1993, 54, 14 - 7 Teicoplanin in home therapy of the terminally ill child; Ball LM et al.; Children discharged in the terminal phase of illness were offered the possibility of having central venous line infections treated with teicoplanin at home by their parents after suitable instruction . The decision to begin antibiotic treatment was subjective, based on a history of rigors and/or raised temperature in an otherwise "well" child . No difficulties were encountered in instructing the chosen parents . In all, five treatment periods of 7 days were required in the five children selected . The review time was 31 weeks (mean duration, 6.2 weeks/patient; range, 4-12 weeks), ended in all cases by death . Infection occurred a mean of 3.2 weeks after discharge (range, 1-8 weeks), and all episodes were successfully treated at home without hospital admission or ward-based support . No deaths occurred as a result of antibiotic therapy failure, and there were no clinically relevant side-effects . Autopsy confirmed the absence of central venous line infection in one patient, but blood culture was positive for Staphylococcus aureus in another . This study shows that home treatment of line infections with teicoplanin is effective and well tolerated, and offers advantages in terms of quality of life and parent-child relationships. J Clin Lab Anal, 1993, 7(4), 209 - 13 Production of hydrogen peroxide by neutrophilic polymorphonuclear leukocytes in patients with diabetic nephropathy; Watanabe A et al.; The production of hydrogen peroxide (H2O2) by neutrophilic polymorphonuclear leukocytes (PMN) after stimulation with phorbol myristate acetate (PMA), n-formyl-l-methionyl-l-leucyl-l-phenylalanine (FMLP), aggregated human IgG, or Staphylococcus aureus was determined in 36 patients with non-insulin dependent diabetes mellitus (NIDDM) . H2O2 production by PMN after stimulation was measured using flow cytometry . Thirty-six patients with NIDDM were divided into four stages as follows: 1) stage I: non-microalbuminuric stage; 2) stage II: microalbuminuric stage; 3) stage III: proteinuric stage without impairment of renal function; and 4) stage IV: proteinuric stage with impairment of renal function . H2O2 production after PMA stimulation in all stages of NIDDM patients was higher than that in healthy controls . This increase of H2O2 production by PMN was particularly observed in stage IV of NIDDM patients after stimulation . Furthermore, H2O2 production in patients in stage IV was higher than that in patients with non-diabetic disease with impairment of renal function . It appears that reactive oxygen species produced by PMN after stimulation under some conditions may play an important role in the progression of diabetic nephropathy. Ann Fr Anesth Reanim, 1993, 12(1), 72 - 4 {Nosocomial Aeromonas hydrophila pneumonia complicating toxic coma}; Poirier T et al.; A 24-year-old male homosexual drug addict was admitted in coma and circulatory failure after a 10 g overdose of acebutolol . The usual resuscitative measures were undertaken, together with administration of adrenaline and gastric lavage . Six hours of external cardiac massage and pacing, and high catecholamine doses (36 mg.h-1 of adrenaline and 60 micrograms.kg-1 x min-1 of dobutamine) were required before the circulatory system became again spontaneously efficient . After this acute episode, the patient improved despite acute tubular necrosis . On the third day, bilateral alveolar and interstitial lesions were found on the chest film . Bronchoalveolar lavage and protected distal brushings were carried out . Both Aeromonas hydrophila and Staphylococcus aureus were found in the cultured brushings . Treatment with ceftriaxone, vancomycin and amikacin was introduced . This nosocomial pneumonia was very haemorrhagic, resulting in several bloody casts responsible for several episodes of atelectasis . The patient was definitely extubated on the 18th day, and left the ICU 23 days later without any sequela . His HIV status was negative . Four other infections with the same strain of Aeromonas hydrophila occurred at the same time as this patient's . The common source for this germ was found to be soft water . Several measures have since been undertaken: removal of a centralized water softener, filtration and higher chlorine content in the water circuit, and updating of intensive care protocols for disinfection of equipment. Acta Neurochir (Wien), 1993, 122(1-2), 130 - 3 Pyogenic parenchymatous and nidus infection after embolization of an arteriovenous malformation . An unusual complication . Case report; Mourier KL et al.; Infectious complications of cerebral angiography and of therapeutic angiographic procedures are very seldom reported . The case of an infected embolized arteriovenous malformation (AVM) by staphylococcus aureus is reported . Abscess formation became manifest seven months after the endovascular procedures . Antibiotherapy was initially started after puncturing the abscess, but finally the cure of the lesion could only be obtained by radical excision of the infected and embolized AVM, as if the persisting embolization material was promoting the infection . The modalities of infection after cerebral endovascular procedures are discussed. Vet Res, 1993, 24(1), 33 - 45 {Modeling of the bactericidal effect of spiramycin . Choice of a pharmacodynamic model}; Renard L et al.; The modelling of in vitro growth curves of Staphylococcus aureus ATCC 6538P at different pHs, of spiramycin killing curves and of the dose-effect relationship was performed . The growth curve was characterised by a Volterra-Kostitzin differential model, the main parameters of which were the bacterial growth rate constant of 0.95 +/- 0.13 h-1 and a lag-time of 0.74 +/- 0.36 h . The Zhi model used for bactericidal kinetics showed the same bacterial killing rate constant from a minimum concentration characteristic of "time-dependent" antibiotics . This differential model incorporated with pharmacokinetic models could lead to a rationale for proper dosage regimens. Med Dosw Mikrobiol, 1993, 45(2), 159 - 62 {Treatment with neomycin and Na2EDTA for skin infections in rabbits caused by Staphylococcus aureus}; Rudy M et al.; The rabbits were infected intradermally with a strain of S . aureus resistant to neomycin . They were treated with neomycin, Na2EDTA and by combination of these substances, which were applied as a cream with differing concentrations of these factors . Simultaneous application of neomycin and Na2EDTA for treatment of experimental staphylococcal infections caused in rabbits with a strain resistant to neomycin, resulted in acceleration of healing of pustular changes, when compared with results obtained in animals treated with these substances alone. Am J Cardiovasc Pathol, 1993, 4(4), 367 - 70 Isolated right-sided endocarditis in Hong Kong Chinese; Dickens P et al.; Thirteen histologically-proven cases of endocarditis confined to the right side of the heart were found in 9406 consecutive autopsies . Eleven cases involved the tricuspid valve and two the atrial endocardium . Nine cases were in males and 4 in females . Only one case showed underlying valvular/endocardial disease . Eight cases were infective, with Staphylococcus aureus being the predominant organism, and 5 non-infective . For the infective cases, intravenous drug abuse was the commonest predisposing factor, and malignancy the commonest association for the non-infective . The commonest overall complication was lung abscess due to septic emboli in the 3 drug addicts, a common association in other reported series . These latter cases are particularly important to recognize, being generally more responsive to treatment than left-sided infective endocarditis. Acta Haematol Pol, 1993, 24(4), 333 - 8 {Production of activity stimulating an increase of clonal granulocytic precursors by normal B lymphocytes under the influence of various mitogens}; Kozlowska-Skrzypczak M et al.; The effect of mitogen-stimulated (concanavalin A, Con A; phytohemagglutinin, PHA; pokeweed mitogen, PWM; Staphylococcus aureus Cowan I, SAC I) normal B lymphocytes on the clonal proliferation of granulocytic progenitors from marrow of healthy subjects (CFU-dG) was studied in diffusion chamber culture . PWM-, SAC- and Con A-stimulated B lymphocytes produced an humoral activity that increased the CFU-dG-derived colony formation . The highest growth-stimulating effect was induced by SAC I-preincubated B lymphocytes and to a lesser degree by PWM- or Con A-stimulated B cells . In contrast, PHA-preincubated and unstimulated B lymphocytes revealed no effect on the CFU-dG proliferation. Surg Today, 1993, 23(10), 880 - 4 The significance of methicillin-resistant Staphylococcus aureus infection in general surgery: a multivariate analysis of risk factors and preventive approaches; Shimada M et al.; To investigate the characteristics, risk factors, and prevention of methicillin-resistant Staphylococcus aureus (MRSA), a total of 3,627 patients were studied . Among these, 1,336 patients with various infections were used in a risk factor analysis of MRSA to determine the relationship between the use of antibiotics and the incidence of MRSA . Only 3.0% of infections were attributed to MRSA, the esophagus and colorectal region being highly involved, as anastomotic or pelvic abscesses, while the lung had a lower incidence . Almost half the patients with MRSA infections (47.6%) had concomitant infection . A univariate analysis revealed the following significant factors: The coexistence of gastrointestinal or metastatic malignancy, sepsis, tracheostomy, and the prior use of antibiotics such as the beta-lactam compounds or aminoglycosides . A multivariate analysis showed that gastrointestinal malignancy, sepsis, and the prior use of aminoglycosides, tetracycline, macrolides, and carbapenems were independently significant factors . To promote the education of doctors and nurses, regular in-service meetings on MRSA were held in the ward . Moreover, preventive approaches such as patient isolation, strategically placed hand washing equipment, and the use of disposable gloves and contaminated waste bags, have been initiated, and the incidence of MRSA has decreased significantly since then . Thus, to control MRSA, the following steps should be taken: (1) constant and careful surveillance, (2) regular risk factor analyses, (3) the optimal administration of antibiotics, and (4) the education of all hospital staff. Microbiol Immunol, 1993, 37(11), 853 - 9 Potent in vitro bactericidal activity of polymyxin B against methicillin-resistant Staphylococcus aureus (MRSA); Yoshida T et al.; Eradication of methicillin-resistant Staphylococcus aureus (MRSA) carried by inpatients or healthy hospital personnel by topical use of antibiotics is an important step for preventing outbreak of MRSA nosocomial infection . In the screening of the antibiotic best suited for this purpose, we have found that polymyxin B, a commonly used antibiotic for gram-negative infection, had an unexpected strong cytokilling activity towards MRSA clinical strains, which was more potent than that of vancomycin or gentamicin . The data suggested that polymyxin B could be an antibiotic of choice in the treatment of topical carriage of or infection caused by MRSA. Medicina (B Aires), 1993, 53(2), 124 - 8 {Evaluation of alternative hemoculture tests for the diagnosis of early neonatal sepsis}; Celadilla ML et al.; In order to evaluate the diagnostic usefulness of coadjuvant tests such as external auditory canal swab culture and cultures from nasopharyngeal and gastric aspirates, and to determine the incidence and etiology of early neonatal sepsis (ENS) at our Unit, 90 newborn cases whose mothers experienced premature rupture of the membranes (PRM) were studied prospectively . Although a firm diagnosis requires positive blood cultures, the difficulty in recovering microorganisms and the trauma induced by sample collection in the baby justify the search for alternative diagnostic tests . Out of 2293 childbirths during 1991, 90 mothers (4%) had PRM more than 24 hours pre-partum, while 6.9/1000 (16/2293) developed ENS . In newborns from PMR mothers, ENS percentage was 3.3%, but increased to 5.5% in association with chorionamnionitis and reached 8.8% in premature cases . Among etiological ENS agents, Gram-positive microorganisms predominated (Table 1), particularly Staphylococcus aureus . Despite the finding that none of the coadjuvant assays (Table 2) had sufficient sensitivity or positive predictive value to identify all septic cases, they may prove useful to pinpoint newborns at high risk due to amniotic fluid exposure to infection or to chorioamnionitis. Surg Today, 1993, 23(11), 947 - 53 Methicillin-resistant Staphylococcus aureus infections following esophageal surgery in patients with impaired defense mechanisms; Saito T et al.; This study was conducted to determine whether or not compromised host defense mechanisms prior to surgery are related to postoperative infections with methicillin-resistant Staphylococcus aureus (MRSA) . Neutrophil cytocidal activities, serum complement and immunoglobulin levels, the in vivo antibody-producing capacity against pneumococcal polysaccharide (PPS), and cell-mediated immunity (CMI) were evaluated in 22 patients who underwent esophagectomy for esophageal cancer between 1989 and 1990 . Postoperatively, nine patients developed MRSA infections . Anti-PPS IgG was found to be significantly lower in patients with MRSA infections than in those without (P < 0.01) . All the patients with MRSA infections showed a titer < 600 EU, while all but one of the non-infected patients showed a titer > 600 EU . Impairment in other components of the defense mechanisms, apart from a partial deficiency of CMI, did not differ between the groups . Thus, a preoperative evaluation of the antibody-producing capacity may serve to predict the development of MRSA-related infections following major surgery such as esophagectomy. J Mol Neurosci, 1993 Fall, 4(3), 161 - 72 Synthesis and characterization of a bioactive 82-residue sphingolipid activator protein, saposin C; Weiler S et al.; The sphingolipid activator protein, saposin C (also termed SAP 2), was chemically synthesized, purified, and characterized . The fully protected 82-residue protein was synthesized by automated solid-phase methods, with multiple recoupling steps resulting in a high average coupling efficiency of 98.8% . The overall yield was estimated to be approx 40% . Deprotection and cleavage of the peptide from the resin was followed by folding in the absence of chaotropic agents at pH 8.5 . The protein was purified by reversed-phase high pressure liquid chromatography (HPLC) and its purity determined by capillary electrophoresis and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) . The composition of the synthetic saposin C was determined by amino acid analysis . Its sequence was verified by Edman sequence analysis of overlapping peptide fragments generated by chymotryptic and Staphylococcus aureus V8 digestions . The sequence at the C-terminus was determined by digestion with carboxypeptidase P, followed by phenylthiohydantoin (PTH) derivitization and HPLC analysis of the released amino acid residues . Deglycosylated native saposin C appeared as a lower molecular-weight species than synthetic saposin C on SDS-PAGE . This has been explained by amino acid and C-terminal analysis showing native saposin C to be two amino acids shorter at the C terminus than a deduced sequence (from cDNA) previously published . Synthetic saposin C displayed 85% of full biological activity as determined by its ability to stimulate glucocerebrosidase activity in vitro: Synthetic and native saposin C increased glucocerebrosidase catalyzed hydrolysis of 4-methylumbelliferyl beta-D-glucoside by factors of 6.0 and 7.1, respectively . Furthermore, synthetic and native saposin C share similar K(act) values (0.5 and 1.5 microM respectively) indicating that they bind to glucocerebrosidase with similar affinities. Postgrad Med J, 1993, 69 Suppl 3, S78 - 83 Action of povidone-iodine against methicillin-sensitive and -resistant cultures of Staphylococcus aureus; Lacey RW et al.; Forty clinical isolates of methicillin-resistant and methicillin-sensitive Staphylococcus aureus were studied for their susceptibility to povidone-iodine (available iodine 11%) under a variety of conditions . The cultures varied in properties, notably in their sensitivity to other therapeutic agents . No differences were observed between the resistant and sensitive cultures in destruction by povidone-iodine . When bactericidal activity occurred, > 99% of the bacterial cells were lethally damaged within 10 seconds of exposure to povidone-iodine . The bacterial cells that had been washed and resuspended thrice in distilled water were most susceptible, with as little as 1 x 10(-16) G (2.36 x 10(5) atoms) of iodine being required to destroy one bacterial cell . Attempts were made to select variants resistant to iodine from the surviving minority population in such experiments . These experiments failed to select resistance, and it is concluded that any apparent variation in vulnerability to povidone-iodine results from aggregation of the bacteria and differences in penetration of povidone-iodine . Substances were studied for their ability to inactivate povidone-iodine . The most potent agents were the free sulphur-containing amino acids, cysteine and methionine . These are present in high concentrations in nutrient broth and most other culture media . This explains the reports of 'pseudo-resistance' to povidone-iodine in certain laboratory experiments . Suggestions are made for the optimum use and formulation of povidone-iodine. Postgrad Med J, 1993, 69 Suppl 3, S66 - 9 Comparison of bactericidal activities of various disinfectants against methicillin-sensitive Staphylococcus aureus and methicillin-resistant Staphylococcus aureus; Yasuda T et al.; Various disinfectants were compared in terms of the duration of bactericidal activity against methicillin-sensitive Staphylococcus aureus (MSSA), and methicillin-resistant Staphylococcus aureus (MRSA), among S . aureus isolated in our hospital . Strains of S . aureus which showed minimum inhibitory concentrations of cloxacillin of less than 1.56 micrograms/ml and of 3.13 micrograms/ml or higher were designated MSSA and MRSA respectively . There was no difference in sensitivity to disinfectants between MSSA and MRSA . There was a great variation in the duration of bactericidal activity of chlorhexidine gluconate against these species with the majority requiring contact times of between 2 minutes and over 20 minutes . All strains except for one strain of MRSA were killed within 20 seconds after disinfection with benzalkonium chloride . All strains were killed within 20 seconds after disinfection with alkyldiaminoethylglycine hydrochloride or povidone-iodine. Postgrad Med J, 1993, 69 Suppl 3, S62 - 5 In vitro efficacy of povidone-iodine solution and cream against methicillin-resistant Staphylococcus aureus; Goldenheim PD; Methicillin-resistant Staphylococcus aureus (MRSA) has emerged worldwide as a significant and difficult-to-treat source of nosocomial infection, emphasizing the ever-present need for reliable and effective antisepsis . Betadine Solution (10% povidone-iodine; PVP-I) and Betadine Cream (5% povidone-iodine) are broad-spectrum microbicides widely used for topical cleansing, wound treatment, and antisepsis . There have been no reports of bacterial resistance to Betadine preparations . In the present study in vitro killing times were determined for PVP-I solution and PVP-I cream against five hospital isolates of MRSA and one methicillin-sensitive strain (MSSA) . Isolates were incubated for 24 hours in trypticase soy broth and 0.5 ml, containing 10(6)-10(7)/ml bacteria, was then removed and added to each test agent containing PVP-I . At intervals of 30 seconds, 1 min, 2 min, 3 min, 5 min, 10 min, and 15 min, 10 microliters aliquots were removed and added to 10 ml of culture broth containing 0.1% sodium thiosulphate, the neutralizing agent for PVP-I . After neutralization the samples were incubated for 48 hours and then observed for growth (turbidity) or its absence . PVP-I solution was effective against all isolates, killing each strain within 30 seconds . PVP-I cream killed four of the MRSA isolates within 30 seconds, and required a 2-3 minute killing time for one strain of MRSA and the strain of MSSA . These results are supported by four published in vitro studies which show that PVP-I is highly effective in killing MRSA . In addition, each study shows that PVP-I is more effective than chlorhexidine against MRSA. Postgrad Med J, 1993, 69 Suppl 3, S27 - 32 Efficacy and safety of a quick drying rubbing type povidone-iodine alcoholic disinfectant solution; Kirita T et al.; The efficacy and safety of a quick drying rubbing type disinfectant (HAD Hand Wash) composed of a povidone-iodine (PVP-I) and ethanol were evaluated in the disinfection of transient skin flora on fingers of 30 medical staff of our department . Three millilitres of HAD solution was rubbed onto the hands for three minutes several times a day over a four week period . Bacteriological effects were evaluated twice, on the first day and after four weeks, by the palm stamp method . The rate of colonies sterilized (bacterial reduction rate) after the use of HAD Hand Wash was 86.9 +/- 21.2% (mean +/- SD) on the first measurement, and 91.5% +/- 12.8% at four weeks . Staphylococcus epidermidis was isolated most frequently and this was eradicated in 83.3% of subjects, the bacterial reduction rate 99.7% . Staphylococcus aureus was originally isolated in 14 subjects and was eradicated in 13; the bacterial colony reduction rate was 99.9% showing high antiseptic efficacy . Although the bacterial reduction rate for Bacillus sp . was a rather low, 75.8-100%, this result was not considered problematic from the clinical standpoint . There were no side effects like chapping in spite of an average 104 applications of HAD Hand Wash during the 4-week test period . From these results, HAD Hand Wash was considered highly effective and safe. Postgrad Med J, 1993, 69 Suppl 3, S126 - 9; discussion S130 Epidemic methicillin-resistant Staphylococcus aureus (EMRSA): experience from a health district of central England over five years; Rahman M; A series of outbreaks of EMRSA occurred in three hospitals of a Health District in the years 1986 and 1987, affecting 64 patients and 6 staff members . By the antibiotic sensitivity pattern (methicillin-resistance and resistance to many others) and phage typing, the organism resembled the epidemic strains involved in London outbreaks . In this series of outbreaks, different circumstances led to adaptation of different control measures with their cost implications . As failure with chlorhexidine was experienced, it was replaced with povidone-iodine for routine handwashing and topical application to patients in affected wards . Heavy environmental contamination involving mattress, bed, floor, table, chair, locker, television etc . was shown . Repeated failures of a phenolic disinfectant led to use of formaldehyde or a higher concentration of the phenolic disinfectant which caused side effects in staff members . Various anti-bacterial agents had been used in treating different conditions in different cases . Mupirocin was found to be the best agent in treating infections or colonization of superficial accessible sites . For non-accessible sites fusidic acid and rifampicin were found to be satisfactory . Continued surveillance in the District for three years after the last case did not detect re-emergence of EMRSA. Postgrad Med J, 1993, 69 Suppl 3, S122 - 5 Efficacy of intranasal application of povidone-iodine cream in eradicating nasal methicillin-resistant Staphylococcus aureus in neonatal intensive care unit (NICU) staff; Masano H et al.; We investigated the staff in our neonatal intensive care unit (NICU) for the presence of methicillin-resistant Staphylococcus aureus (MRSA) in the nasal cavity, and then applied intranasal povidone-iodine cream to the physicians and nurses working on the unit . Prior to the application of povidone-iodine cream, the isolation rate of S . aureus from the nasal cavity was 30% for the physicians and nurses in contact with NICU patients (contact group), not significantly different from the 33.3% rate for other hospital staff (control group) . The isolation rate for the contact group decreased to 10.5% after application of the cream . Although MRSA was not isolated from the nasal cavity of those in the control group, it was isolated from 13.3% of those of the contact group before application of the cream . Nasal MRSA disappeared after use of the cream . No adverse reactions or abnormalities in serum levels of thyroid hormone-related substances were observed in any of the subjects . These results indicated that the nasal application of povidone-iodine cream is safe and effective for eradicating MRSA in the nasal cavity. Postgrad Med J, 1993, 69 Suppl 3, S117 - 21 Prevention and control of nosocomial infection caused by methicillin-resistant Staphylococcus aureus in a premature infant ward--preventive effect of a povidone-iodine wipe of neonatal skin; Aihara M et al.; In early 1983 we experienced a small scale epidemic of Staphylococcus aureus coagulase type IV in the premature infants unit . Children had bacteraemia or impetigo . The microorganism was resistant to methicillin, erythromycin and lincomycin and was susceptible to tetracycline, chloramphenicol and cefmetazole . The results of coagulase typing and antimicrobial sensitivities indicated that these cases represented nosocomial infection with MRSA . The source and route of the infection were investigated, and measures were taken to prevent bacterial spread from carriers and to keep instruments and environments clean . As the source of infection was not identified, we tried wiping the body surface of the premature infants with a diluted IsodineR solution (10% povidone-iodine; 1:100 dilution) in order to prevent colonization of the microorganism on the body surface . As a result, no additional MRSA infection occurred in the premature infant unit . During the subsequent 6 years of frequent surveys of carriers and wiping the appropriate body surface with diluted IsodineR solution we have had no recurrence of MRSA . None of the premature infants wiped with IsodineR solution showed any objective abnormalities, although laboratory testing disclosed an elevated blood iodine level and a transient mild reduction of T4 in some infants. Postgrad Med J, 1993, 69 Suppl 3, S106 - 16 Clinical significance of methicillin-sensitive and methicillin-resistant Staphylococcus aureus in UK hospitals and the relevance of povidone-iodine in their control; Gordon J; This review summarizes the natural history, clinical relevance and basis of control of Staphylococcus aureus infection in UK hospitals, stressing the central role of asymptomatic carriage by patients and staff in persistence of this prolific and versatile nosocomial pathogen . The clinical relevance of methicillin-resistant and methicillin-sensitive S . aureus (MRSA and MSSA) is considered in terms of prevalence and spectrum of invasive and toxigenic infections produced, correlated with host and parasite risk factors . An assessment is made of arguments why the acquisition of methicillin-resistance or multiple antibiotic resistance might justify more than conventional methods of containment and how the control policy is influenced by the expression of enhanced virulence and epidemicity . Guidelines for control of epidemic MRSA (EMRSA) are discussed with reference to justification, feasibility and efficacy . As elimination of carriage is crucial to the success of any rational control policy the relative merits of topical antibiotics and antiseptic agents are compared . The bacterial efficacy of povidone-iodine, chlorhexidine and mupirocin are evaluated as a basis for eradication of MRSA. Scand J Infect Dis, 1993, 25(5), 647 - 53 Changing pattern of phage group II Staphylococcus aureus infections: from community to hospital; Faber M et al.; Changes in Staphylococcus aureus strains of phage group II were evaluated over a 30-year period . Strains of this group were previously known to be mainly community-acquired, to be rather sensitive to antibiotics and to occur in infections with better clinical outcome . During the period 1961 to 1990 a total of 551, 563 S . aureus strains was characterized according to antibiotic resistance, phage type and site of isolation . Clinical information was obtained on 16,781 bacteraemia cases . Strains of group II occurred until 1983 with a rather constant frequency around 16% of the S . aureus population . Since then, a constant increase in frequency has taken place and since 1990 they are the most frequently isolated phage pattern (22%) . Bacteraemia caused by these strains occurs today with the same frequency in hospital and community as strains of other phage patterns . Moreover, these strains have changed from being highly susceptible, to being more resistant to penicillins and tetracycline than the rest of the population . Resistance to methicillin remained low in group II strains throughout the period (about 0.2%) . Mortality rate and the proportion of patients with underlying diseases have been the same in recent years regarding group II strains as with other strains . This study indicates that the previous distinction between community-acquired and hospital-acquired S . aureus infections is no longer valid. Microbiol Immunol, 1993, 37(9), 695 - 703 A factor found in aged tungstate solution enhanced the antibacterial effect of beta-lactams on methicillin-resistant Staphylococcus aureus; Tajima Y et al.; We have found a factor (Factor T) in aged mixtures of tungstate and phosphate which greatly enhances the antibacterial effects of beta-lactams on both inducible and constitutive methicillin-resistant Staphylococcus aureus, but not on methicillin-susceptible S . aureus . Factor T alone did not strongly inhibit bacterial growth . There was no synergism of Factor T with other classes of antibiotics, nor with other groups of bacteria, and it reduced the efficacy of amino-glycosides and tetracycline . Upon preparation of Factor T, acidifying and heating the mixture of tungstate and phosphate resulted in a high yield and reproducibility, and no enhancing effect was observed when other anions such as sulfate or molybdate were used instead . Factor T is heat- and acid-stable but labile to alkalization, and is probably a complex of phosphate and tungstate. Microbios, 1993, 76(306), 19 - 28 Cell wall-active antibiotics induce specific alterations of the electrophoretic profile of membrane proteins in a Staphylococcus aureus strain; Najioullah F et al.; The effect of six representative cell wall-active antibiotics upon the membrane proteins of Staphylococcus aureus CIP 65-25 was studied . The profiles of whole membrane proteins and of penicillin-binding proteins of strain CIP 65-25 grown on an antibiotic-free medium were analysed and compared with those of nine other S . aureus strains . Both profiles were found to be very similar for all strains examined . Cell wall-active antibiotics brought about definite changes dealing with a few membrane proteins of strain CIP 65-25 . Some polypeptide bands of the control profiles decreased or vanished after exposure to antibiotics whereas others rose only in antibiotic-treated bacteria . None of these modified polypeptides was a penicillin-binding protein . Their possible involvement in the antibacterial effects of antibiotics or in the determinism of bacterial resistance to antibiotics is discussed. Nord Med, 1993, 108(11), 286 - 8 {Hvidovre Hospital . Measures to control the spread of imported multiresistant Staphylococcus aureus}; Kolmos HJ; Methicillin-resistant Staphylococcus aureus (MRSA) are endemic in many hospitals abroad . Patients transferred from these hospitals may give rise to a spread of MRSA in Scandinavian hospitals, where MRSA only occur sporadically . A spread of MRSA may have great therapeutic and economic consequences for the local health care system and should therefore be met by strict hygienic measures . In Hvidovre Hospital patients transferred from or recently admitted to hospitals outside Scandinavia, are screened for MRSA . Patients carrying MRSA are isolated and discharged as soon as possible . Attempts are made to eliminate the carrier state by means of chlorhexidine. Med Dosw Mikrobiol, 1993, 45(1), 37 - 40 {Modulation of lymphocytes and immunoglobulins by serine proteinase of Staphylococcus aureus}; Porwit-Bobr Z; Serine proteinase of S . aureus binds with surface of peripheral blood lymphocytes and by proteolysis of polypeptides on the membrane and transduction of signals, changes in polyclonal activation of T and B lymphocytes occur . Cleavage of immunoglobulins tested in three classes and evaluated by electrophoresis in polyacrylamide gel (SDS-Page) of Ig fragments, was differing . IgG and IgG fragments were degraded in higher degree than IgM, H and L chains or i Fc and i Fab fragments . Influence of proteolytic modulation of IgG particles on fixation antigenic fractions homogenized by ultrasound cells of S . aureus was evaluated in ELISA test . Basing on detailed results postulated mechanism of modification effect of immunoglobulins and lymphocyte reaction under influence of serine proteinase of S . aureus in microenvironment of after-inflammatory reaction in staphylococcal infection, has been accepted. Med Dosw Mikrobiol, 1993, 45(1), 19 - 23 {Increase of pathogenicity of Staphylococcus aureus strains caused by lysogenic conversion by phages of serologic group F}; Mlynarczyk G et al.; The substantial influence of presence of the double converting prophages of the serological group F on the decrease of intensity of human leukocytes stimulation with lysogenic cells of S . aureus was determined . The end values of chemiluminescence in the six experimental systems containing lysogenic bacteria were only from 15.4 to 37.2% of the value in the control sample . The end values of the intercellular killing of the two lysogenic variants were twofold lower than in controls . Third of the investigated variants was completely resistant to intracellular killing in the applied system, and revealed 27% of growth stimulation. Jpn J Ophthalmol, 1993, 37(2), 165 - 70 Ophthalmic diseases in bedridden patients with severe dementia; Kiuchi Y et al.; Ophthalmologic examinations were performed on the 176 eyes of 88 bedridden patients with severe dementia . There were some pathologic changes in the external and anterior segments of 93 eyes . We observed infectious external and anterior segment eye diseases in 80 of these 93 eyes (86.0%) . Fundus diseases were found in 34 eyes, but treatment was not necessary . Infectious eye diseases appeared to be the most prevalent ocular problem in these patients . Cultures of conjunctival swabs commonly demonstrated Staphylococcus aureus . No differences were noted in tear secretion rate or lysozyme concentrations in tears between eyes with and without infection . The blinking frequency was significantly reduced in infected eyes compared to normal eyes . Failure of the lacrimal drainage system was observed more often in infected eyes than in normal eyes . This study demonstrated that bedridden patients with severe dementia develop infectious eye diseases easily, to which stasis of tear flow seems to predispose . In caring for bedridden patients with severe dementia, ophthalmologists must be concerned with controlling infectious eye diseases. Drugs Exp Clin Res, 1993, 19(2), 59 - 64 Microbicidal effects of ozone solution on methicillin-resistant Staphylococcus aureus; Yamayoshi T et al.; Methicillin-resistant Staphylococcus aureus has become an important problem in chemotherapy in the past decade because of its resistance to commonly used disinfectants . The usefulness of ozone solutions to disinfect against strains of this bacterium has been evaluated . Ozone strongly oxidizes the cell walls and cytoplasmic membranes of bacteria . The microbicidal effect of ozone solutions took place within the first five seconds of treatment . An ozone concentration 1.5 times that needed to kill the one methicillin-sensitive strain tested was sufficient to kill the resistant clinical isolates . Such an ozone solution could be used to disinfect medical instruments and similar equipment . Ozone solutions should be useful in reducing the number of bacterial infections caused by inadequate disinfection against new resistant strains. Respiration, 1993, 60(3), 182 - 5 Conservative treatment of empyema in children; Gocmen A et al.; Seventy-two children with no underlying diseases were treated for empyema . Radical surgical approaches like decortication were necessary for only 3 children . In 66 children tube drainage was applied . Staphylococcus aureus was cultured or was shown in Gram's stain in 32 (44%) and children with this microorganism had longer duration of tube drainage (p < 0.05) . The period for normalization of chest X-ray was positively related with the age of the patient (p < 0.05) . In 60 of 72 children, chest X-ray was normal after 3 months . Follow-up of the patients 18 months after the infection revealed that pulmonary radiograms were normal in all cases and pulmonary function tests were within normal limits in all of the tested children (n = 25) . It is emphasized that avoiding major surgical approaches must be encouraged in childhood empyema. Rev Med Interne, 1993, 14(7), 733 - 6 {Staphylococcus aureus endocarditis: an unusual clinical picture}; Acar P et al.; We describe an unusual presentation of bacterial endocarditis due to Staphylococcus aureus in a 50 year-old woman with mitral insufficiency . The disease began by a vascular purpura without fever and a digital embolism . The source of infection was anal ulcerations . Diagnosis of endocarditis was made possible by trans-oesophagus echocardiography but not by trans-thoracic echocardiography . The patient was successfully treated by surgery associated with antibiotherapy . This observation emphasizes the indications of surgery at the acute phase of endocarditis . The anal source of endocarditis is original, no other case has been found in the literature. Acta Microbiol Hung, 1993, 40(2), 91 - 9 Multiple effects of promethazine in Staphylococcus aureus; Molnar J et al.; The antibiotic resistance of 6 Staphylococcus aureus strains was eliminated with a frequency from 1.2 to 10% in the presence of subinhibitory concentrations of promethazine . The pigment production of the cells was also eliminated by the promethazine to an extent of 0 to 5% . The cell size was increased and the protein A production was markedly decreased in S . aureus cells cultured in the presence of promethazine . Complex formation between protein A and promethazine was detected by differential spectrophotometry . The biological activity of staphylococcus protein A was abolished by promethazine in the passive haemagglutination of rabbit antiserum treated sheep red blood cells . Evidence has been found that plasmid-encoded functions of S . aureus cells can be altered in the presence of promethazine, and the chromosomally controlled synthesis of protein A, one of the weakest virulence factor of S . aureus is also lowered by promethazine. Am J Nephrol, 1993, 13(6), 435 - 6 Retropharyngeal infection with Staphylococcus aureus in a haemodialysis patient; Hughes J et al.; The case of a 55-year-old female Asian haemodialysis patient is described . She developed a septicaemic illness and retropharyngeal soft-tissue infection secondary to Staphylococcus aureus infection. J Mal Vasc, 1993, 18(4), 327 - 30 {Aortic thrombosis during acute endocarditis caused by Staphylococcus aureus}; Pistorius MA et al.; The authors report a case of acute bacterial (Staphylococcus aureus) endocarditis in a 70-year-old woman, revealed by a febrile cerebral ischemic accident . Ultrasonography confirmed the presence of a large posterior mitral valve vegetation interfering with left ventricular filling . The sudden onset of complete paraplegia and acute ischemia of the lower limbs suggested thrombosis of the abdominal aorta, which was confirmed by aortography . These features indicated that a vegetation fragment had migrated, obstructing the aortic bifurcation and causing secondary thrombosis . This led in turn to involvement of the medullary arteries and the onset of paraplegia . Unfortunately, acute renal insufficiency and major left heart failure rapidly developed, and the patient died . Autopsy confirmed the diagnosis of aortic thrombosis with involvement of the renal arteries . Multiple visceral infarcts were noted as well as the large mitral vegetation . This case illustrates the potential severity of systemic embolism complicating endocarditis due to Staphylococcus aureus . The accident was remarkable because of the aortic acute occlusion and the association with paraplegia, an unusual neurologic complication. Trop Geogr Med, 1993, 45(6), 294 - 6 Bacterial pathogens in malnourished children with pneumonia; Fagbule DO; Lung puncture aspirates were obtained from 99 malnourished children aged 9 months to 5 years with pneumonia, in order to identify the causative bacterial agents . Klebsiella species, Staphylococcus aureus and Escherichia coli were identified in 39.4%, 30.3% and 8.8% of aspirates, respectively . Approximately half to two-thirds of the children in each aetiological group had the same organisms isolated in both lung aspirates and blood . The bacterial agents reflected the immunocompromised status of the patients . Isolation of Klebsiella species was associated with age (p < 0.01), and type of malnutrition (p < 0.05); while that of Staphylococcus aureus was only statistically associated with type of malnutrition (p < 0.01) . Gentamicin and cloxacillin represent a safe antibiotic regime for pneumonia in children with moderate to severe malnutrition. J Ocul Pharmacol, 1993 Winter, 9(4), 311 - 20 Bioassays for quantitating ciprofloxacin and tobramycin in aqueous humor; Engel LS et al.; Development of new therapies for bacterial keratitis requires assays of antibiotics in infected ocular tissues; however, methods for such drug quantitation can be expensive and can lack precision and sensitivity . To develop a more suitable assay for quantitating tobramycin and ciprofloxacin in the aqueous humor, sixteen bioassays were compared . Variables assessed included: the strain of bacteria used as indicator (Staphylococcus aureus ATCC 25923 versus Klebsiella pneumoniae ATCC 10031), the application of bacteria to the agar surface versus inclusion in molten agar, the volume of the agar, and the application of antibiotic on paper discs versus instillation into agar wells . The best results were obtained when the appropriate indicator organism (S . aureus for tobramycin and K . pneumoniae for ciprofloxacin) was applied to the surface of the agar, antibiotic solution was put into agar wells, and the volume of agar was 30 ml per plate . The best sensitivity for assays of tobramycin and ciprofloxacin diluted in water were 0.98 and 0.12 micrograms/ml, respectively . These sensitivities improved as much as fourfold when antibiotics were diluted in aqueous humor or normal saline . The precision of all assays tested was high as determined by regression coefficients (r > 0.90) . The sensitivity and precision of these assays indicate their utility for quantitating fluoroquinolones and aminoglycosides in aqueous humor. Adv Perit Dial, 1993, 9, 248 - 50 Hypertonic saline compresses: therapy for complicated exit-site infections; Strauss FG et al.; We review our experience with hypertonic saline compress therapy in 17 patients with complicated peritoneal dialysis catheter exit-site infections (ESIs) . Compresses consisted of exit-site application of 4-5 gauze pads soaked with warm 3% saline for 5-10 minutes, three times daily, for 2-4 weeks, followed by once-daily use thereafter . The mechanism of action involves inhibition of bacterial growth by a hypertonic medium . Eleven patients with cultures positive for Staphylococcus aureus or Pseudomonas were treated with local exist-site measures (cleansers, antiseptics, antibiotic ointments) . Therapy, which included multiple courses of systemic antibiotics, failed in 8 patients; in 3 patients, who were intolerant to antibiotics, ESI remained unresolved after local care only . Six patients with culture-negative ESIs received no systemic antibiotics and were unimproved following local therapy . Factors associated with therapy failure included malnutrition, diabetes, obesity, and dermal sensitization and injury associated with prolonged topical agent use . Following hypertonic saline compress therapy, we observed resolution of ESI in all patients without recurrence for follow-up intervals of 3-12 months (mean 6.5 months) . Advantages of this therapy include excellent patient acceptance, ease of use, lack of adverse effects on exit site, adjacent skin, catheter or systemic reaction, and minimal expense . Future potential applications include routine daily use for infection prophylaxis and as therapy combined with antibiotics for established ESIs. Adv Perit Dial, 1993, 9, 236 - 9 Breaking-in after the inse