J Vet Med Sci, 1995 Apr, 57(2), 351 - 3 Application of the Limulus amoebocyte lysate test as an indicator of microbial contamination in pork carcasses; Misawa N et al.; The Limulus amoebocyte lysate (LAL) test, a simple and rapid method, was applied for the evaluation of bacterial contamination of carcasses in a slaughterhouse . Twenty-five pork carcasses were examined for bacterial contamination, and we evaluated the correlation between the LAL test and the methods usually employed for detecting indicator organisms of total viable cells, coliforms or Staphylococcus aureus . The reciprocal of the highest dilution found to be positive was used as the LAL index . A high correlation was observed between the LAL index and the number of coliforms, but not between the index and the number of other organisms . The LAL test was accomplished within 2 hr after sampling, and thus may serve as a useful tool for the substitute indicator of coliforms among bacterial contamination in carcasses. J Vet Med Sci, 1995 Apr, 57(2), 299 - 305 Proliferative response and cytokine production of bovine peripheral blood mononuclear cells induced by the superantigens staphylococcal enterotoxins and toxic shock syndrome toxin-1; Yokomizo Y et al.; The potential of staphylococcal enterotoxin A (SEA), B (SEB), C(SEC) and toxic shock syndrome toxin-1 (TSST-1) to act as superantigens by inducing polyclonal T-cell mitogenesis and cytokine production was tested on bovine peripheral blood mononuclear cells (PBMC) . These four toxins were capable of inducing strong proliferative response of PBMC from calves over a broad dosage range (1 pg/ml to 1 microgram/ml) in vitro . The toxin-activated blast cells consisted of both CD4+ T-cells and CD8+ T-cells, but the T-cell proliferation depended upon the presence of monocytes . Treatment of monocytes with monoclonal antibody to major histocompatibility complex class II antigens substantially inhibited the toxin-induced T-cell proliferative response, but paraformaldehyde-fixation did not abrogate the accessory function . SEA, SEB, SEC and TSST-1, all induced the in vitro release of interleukin-2, interferon gamma and tumor necrosis factor alpha in a dose dependent manner . The results indicate that SEA, SEB, SEC and TSST-1 are capable of acting as superantigens by stimulating bovine T-cells as shown in the human and murine systems . The possible implications of these toxins in the immunopathogenesis of bovine mastitis caused by the infection with Staphylococcus aureus are discussed. Zentralbl Veterinarmed B, 1995 Apr, 42(2), 118 - 26 Effects of Staphylococcus aureus mastitis after endotoxin application on milk yield and composition during subsequent lactation of guinea-pigs; Vandeputte-Van Messom G et al.; The effects of Staphylococcus aureus mastitis on milk yield and composition throughout subsequent lactation in lactating guinea-pigs and the role of endotoxin pretreatment on these phenomena were investigated . Primiparous lactating guinea-pigs were intramammarily inoculated with sterile saline (group 1), S . aureus strain UC 6097 (group 2), or with S . aureus UC 6097 after endotoxin pretreatment (group 3) . Clinical signs and survival rate were monitored . During the second lactation, daily milk yield was measured and milk composition was determined . In primiparous inoculated guinea-pigs, moderate (group 3)-to-severe acute mastitis (group 2) was produced . During subsequent lactation, milk yield in the control group peaked on day 5 and then decreased . Concentrations of Na+ and Cl- in milk, and concentrations of fat, gradually increased, but lactose and K+ decreased . After an early decrease, NAGase in milk increased towards the end of lactation . Except for higher NAGase concentrations in group 3, milk yield and composition during the second lactation did not differ significantly between the mastitis and the control groups . Endotoxin pretreatment only plays a role in the determination of the severity of the infection. FEBS Lett, 1995 Mar 27, 362(1), 80 - 4 Overproduction of the bleomycin-binding proteins from bleomycin-producing Streptomyces verticillus and a methicillin-resistant Staphylococcus aureus in Escherichia coli and their immunological characterisation; Sugiyama M et al.; The bleomycin-binding proteins designated BLMA and BLMS, which confer resistance to bleomycin (Bm), from Bm-producing Streptomyces verticillus ATCC15003 and a methicillin-resistant Staphylococcus aureus B-26, respectively, were overexpressed in Escherichia coli . The present study showed that both BLMA and BLMS quench the antibacterial activity of Bm by the binding to the drug . To immuno-characterize the Bm-binding proteins, we constructed a monoclonal antibody against BLMA . The antibody, designated 893-12, did not cross react to BLMS and another Bm-binding protein from tallysomycin-producing Streptoalloteichus hindustanus . Although the ability of Bm to cleavage DNA was eliminated by a binding of BLMA to Bm, as shown by Sugiyama et al . {Gene 151 (1994) 11-15}, the Bm-induced DNA degradation was restored by pre-incubation of BLMA with the anti-BLMA monoclonal antibody. Nippon Naibunpi Gakkai Zasshi, 1995 Mar 20, 71(2), 167 - 72 {A rare case of a diabetic patient with small cell lung cancer, initially diagnosed as pyogenic vertebral osteomyelitis}; Miyamoto T et al.; A rare case of a patient with non-insulin-dependent diabetes mellitus (NIDDM) with small cell lung cancer, initially diagnosed as pyogenic vertebral osteomyelitis, was reported . A 40-year-old male patient was diagnosed with NIDDM about 3 years earlier, but he did not receive any treatment . Then, a two-month history of high fever, persistent cough and back pain developed . Chest X-ray film showed a lung infiltrate with a small cavity in the upper portion of the left lung . Computed tomography and magnetic resonance imaging of the chest revealed a tumor mass shadow with osteoclasia along the bodies of the 6th and 7th thoracic vertebral bone . Staphylococcus aureus infection was confirmed by arterial blood culture . Administration of antibiotics resulted in the disappearance of the left lung infiltrate and a slight reduction of the tumor mass in the thoracic vertebral bone, suggesting pyogenic vertebral osteomyelitis as an unusual complication of NIDDM . However, as the tumor mass still remained, needle biopsy for the mass lesion was performed, resulting in the diagnosis of metastasis of small cell carcinoma from the left lung . Gene aberration in this lung disease has been reported recently, and its correlation with NIDDM which may also be induced by genetic abnormality is an interesting question that remains to be resolved. Eur J Pharmacol, 1995 Mar 15, 289(1), 59 - 66 Ca2+ handling mechanisms underlying neuropeptide Y-induced contraction in canine basilar artery; Tanaka Y et al.; The effects of neuropeptide Y on isometric tension simultaneously measured with cytosolic Ca2+ concentration ({Ca2+}cyt) and Ca2+ sensitivity of contractile elements were studied in isolated canine basilar arteries . Neuropeptide Y (1-100 nM) increased {Ca2+}cyt and tension in a concentration-dependent and parallel manner, whereas 9,11-dideoxy-11 alpha,9 alpha-epoxymethano prostaglandin F2 alpha (U46619) (10-100 nM), a thromboxane A2 mimetic, produced a large contraction with a small increase in {Ca2+}cyt . Ca2+ channel antagonists such as d-cis-diltiazem (10 mM) abolished both {Ca2+}cyt and tension augmented by neuropeptide Y . In Ca(2+)-free solution containing 0.2 mM EGTA, neuropeptide Y did not change {Ca2+}cyt and tension, whereas U46619 transiently increased both of them . Furthermore, neuropeptide Y apparently did not affect the Ca2+ sensitivity when assessed in the artery permeabilized with Staphylococcus aureus alpha-toxin, whereas U46619 augmented it . These findings suggest that neuropeptide Y-induced contraction in the canine basilar artery is produced mainly by Ca2+ influx through L-type Ca2+ channels. Eur J Biochem, 1995 Mar 15, 228(3), 798 - 804 Lactose-specific enzyme II of the phosphoenolpyruvate-dependent phosphotransferase system of Staphylococcus aureus . Purification of the histidine-tagged transmembrane component IICBLac and its hydrophilic IIB domain by metal-affinity chromatography, and functional characterization; Peters D et al.; The lactose-specific integral-membrane-protein enzyme II (IICBLac) of the bacterial phosphoenolpyruvate-dependent phosphotransferase system of Staphylococcus aureus catalyses the uptake and phosphorylation of lactose . It consists of an N-terminal membrane-spanning IIC domain and a C-terminal hydrophilic IIB domain . IICBLac was fused with a C-terminal tag of six histidine residues using recombinant DNA technology . The resulting protein, IICBLac-His, was produced in Escherichia coli and purified under nondenaturing conditions to homogenity . The purification procedure consists of a NaOH extraction step followed by solubilisation with Triton X-100, and metal-affinity chromatography using Ni(2+)-nitrilotriacetic acid resin . The purified recombinant His-tagged protein possessed substrate specificity identical to that of the wild-type protein . To investigate the hydrophilic IIB domain, the DNA sequence coding for IIB and the His tag were fused in-frame to a DNA sequence specific for an initiation signal . The overproduced recombinant IIBLac-His was obtained by metal-affinity chromatography in pure form . Bacterial phosphotransferase-system-dependent phosphorylation of IIB-His was demonstrated in a photometric assay and by urea/polyacrylamide gel electrophoresis . The phosphorylation activity of the mutant protein {C476S}-IICBLac, containing the mutagenized phosphorylation site, was restored in the presence of IIBLac-His in a phosphorylation assay. Eur J Biochem, 1995 Mar 15, 228(3), 772 - 8 The DNA-binding site of the RecA protein . Photochemical cross-linking of Tyr103 to single-stranded DNA; Morimatsu K et al.; To investigate the DNA-binding site in the Escherichia coli RecA protein, RecA was covalently cross-linked to oligodeoxythymidine {p(dT)14} by irradiation with ultraviolet light . We identified the site of cross-linking of the protein when the RecA.p(dT)14 complex was formed in the absence of nucleotide cofactor as well as in the presence of adenosine 5'-{gamma-thio}triphosphate . When RecA.p(dT)14 complex formed without nucleotide cofactor was irradiated with ultraviolet light, a cross-linked peptide was found after digestion with Achromobactor lyticus protease I . Amino acid composition of the peptide was determined . The results indicated that the site of cross-linking was in the region spanning amino acid residues 89-106 . Further digestion of the cross-linked fragment with Staphylococcus aureus V8 protease indicated that Tyr103 was the site of cross-linking . When the complex formed with adenosine 5'-{gamma-thio}triphosphate was irradiated with ultraviolet light, two cross-linked sites were detected, which were in the region of residues 89-106 and residues 178-183 . These regions are far from the two disordered loops in the crystal structure, which were suggested to be DNA-binding sites by Story et al . {Story, R . M., Weber, T . W . & Steitz, T . A . (1992) Nature 355, 318-325}. Eur J Biochem, 1995 Mar 15, 228(3), 732 - 8 Comparative biochemical and molecular analysis of the Staphylococcus hyicus, Staphylococcus aureus and a hybrid lipase . Indication for a C-terminal phospholipase domain; Nikoleit K et al.; The lipase gene, geh, from Staphylococcus aureus NCTC8530 was cloned in Staphylococcus carnosus . DNA sequencing revealed an open reading frame (ORF) of 2046 nucleotides encoding a 682-amino-acid protein with a molecular mass of 76900 Da . Determination of the transcriptional start site revealed a 203-nucleotide mRNA leader . Expression of geh in the protease-negative S . carnosus (pT181copSA22) resulted in overexpression of a 83-kDa lipase found in the culture supernatant . N-terminal protein sequencing and sequence comparison with three other staphylococcal lipases suggest that this lipase is organised as a pre-pro-enzyme . The substrate specificity of this lipase is different from the Staphylococcus hyicus lipase . The S . hyicus lipase expressed both a high Ca(2+)-dependent phospholipase and lipase activity while the S . aureus lipase lacked this phospholipase activity and its activity with tributyrylglycerol or p-nitrophenyl octanoate is hardly stimulated by Ca2+ ions . A hybrid protein was constructed in which the C-terminal 146 residues of the S . hyicus lipase were substituted by 145 residues of the C-terminal of the S . aureus lipase, which contains the proposed active-site amino acids Asp602 and His641 . The hybrid enzyme was still active and revealed an intermediary enzymic activity . The most striking effect was that it had lost the S . hyicus-specific phospholipase activity and that, in contrast to the two parental enzymes, its activity with p-nitrophenyl octanoate became highly sensitive to the presence of Ca2+ . These observations suggest that the C-terminal domain of the S . hyicus lipase strongly contributes to the binding pocket of the polar headgroup of phospholipids . The Ca(2+)-binding site seems to be located in the N-terminal fragment of the S . hyicus lipase . The fact that two closely related enzymes differ in the need for Ca2+ underscores the notion that it plays a structural rather than a catalytic role. Spine, 1995 Mar 15, 20(6), 685 - 8 The effect of prophylactic antibiotics on iatrogenic intervertebral disc infections . a rabbit model; Guiboux JP et al.; STUDY DESIGN . A rabbit model was used to test the efficacy of two commonly used prophylactic antibiotics, cefazolin and vancomycin, in preventing iatrogenically introduced Staphylococcus aureus intervertebral disc infections . OBJECTIVE . This study was performed to assess the efficacy of two prophylactic antibiotics in preventing iatrogenically introduced Staphylococcus aureus intervertebral disc infections . SUMMARY OF BACKGROUND DATA . Previous studies have had conflicting results regarding the penetration of antibiotics into the nucleus pulposus and their ability to eradicate infection . METHODS . In this study, 40 adult New Zealand White rabbits underwent inoculation of 10(1) or 10(3) Staphylococcus aureus/ml into 3-6 lumbar intervertebral discs under direct visualization . Either no antibiotics (control groups) or various preoperative and postoperative dosing schedules of cefazolin or vancomycin were given intravenously . Five days after surgery, the discs were harvested and cultured . RESULTS . All 40 discs inoculated in the control groups became infected . None of the 35 discs inoculated in the cefazolin groups became infected . Infection developed in 23 of 107 discs inoculated in the vancomycin groups . Most notable of these were 17 of 17 positive cultures in animals given vancomycin 8 hours preoperatively only . CONCLUSIONS . Based on these results, it was concluded that intravenous cefazolin or vancomycin given within 1 hour before surgery can effectively prevent postoperative discitis . No advantage was found with additional postoperative antibiotics. Southeast Asian J Trop Med Public Health, 1995 Mar, 26(1), 78 - 85 An unusual outbreak of food poisoning; Thaikruea L et al.; On August 25 1990, over 400 people who attended a Thailand handicappeds' sport day at a provincial physical education college developed gastrointestinal symptoms after having dinner . An epidemiological team want to determine causes(s) and recommend how to prevent and control a food poisoning outbreak . The investigation included interviewing all 1,210 persons who attended the sport's day . In addition, an environmental survey, laboratory analysis of food samples, and rectal, ear, throat and nasal swabs from foodhandlers were also performed . A case was defined as a person who ate any items of dinner food and experienced vomiting, nausea, abdominal pain, and diarrhea . There were 485 cases out of 1,094 persons, an attack rate of 43% . Interviews were completed for 470 out of 485 cases . The three most common symptoms were nausea (93%), vomiting (88%), and abdominal pain (81.5) . The mean incubation period was 3.20 hours . Three out of four items of food had a significant association with illness . Among these 3 items, eclairs had to the highest crude relative risk, 7.0 (95% CI = 4.8, 10.2) . For statistical analysis, logistic regression by unconditional method was used, and found that only eclairs which were prepared during the night before the dinner and kept at room temperature for at least 12 hours before serving, remained statistically significant in the model (RR = 11.96; 95% CI = 9-22) . Laboratory examination of foods and foodhandlers indicated heavy growth of Staphylococcus aureus producing toxins A and C and Bacillus cereus in eclairs . Culture of nasal swabs from healthy foodhandlers identified B . cereus and S . aureus of different phage types from those in eclairs.(ABSTRACT TRUNCATED AT 250 WORDS) Am J Physiol, 1995 Mar, 268(3 Pt 2), H1223 - 31 Effects of cGMP on calcium handling in ATP-stimulated rat resistance arteries; Andriantsitohaina R et al.; The mechanisms by which guanosine 3',5'-cyclic monophosphate (cGMP) modulates the contraction induced by ATP were investigated in small mesenteric resistance arteries of the rat . The nitric oxide donors 3-morpholinosydnonimine (SIN-1, 10 microM) and sodium nitroprusside (SNP, 10 microM) increased cGMP but not adenosine 3',5'-cyclic monophosphate (cAMP) content of the tissue . SIN-1, SNP, and 8-bromoguanosine 3',5'-cyclic monophosphate (8-BrcGMP, 100 microM) inhibited the myosin light chain phosphorylation and the contractile response to ATP . Both effects were completely reversed by the selective inhibitor of cGMP protein kinase, Rp-8-bromoguanosine 3',5'-cyclic monophosphorothioate (30 microM) . The sensitivity to Ca2+ of arteries permeabilized with Staphylococcus aureus alpha-toxin (4,000 hemolytic units/ml) was not affected by 8-BrcGMP . The two nitric oxide donors and 8-BrcGMP decreased the rise in intracellular Ca2+ induced by ATP . The vasodilator agents abolished the contractile response to the exogenous calcium in vessels that were exposed to 3 mM ATP after depletion of intracellular Ca2+ stores . Thapsigargin (1 microM), an inhibitor of the sarcoplasmic reticulum Ca(2+)-adenosinetriphosphatase, reversed the inhibitory effect of the vasodilator agents when the contraction induced by ATP was elicited in the presence of the Ca2+ entry blocker nitrendipine (1 microM) or in Ca(2+)-free medium . These results show that cGMP inhibits ATP-induced contraction by decreasing intracellular Ca2+ concentration in small resistance arteries . They indicate that this effect results from decreased Ca2+ influx and enhanced Ca2+ sequestration through a thapsigargin-sensitive pump via activation of a cGMP protein kinase. J Lab Clin Med, 1995 Mar, 125(3), 356 - 69 Immunosuppressive properties of surfactant and plasma on alveolar macrophages; Allen JN et al.; Alveolar macrophages have been shown to be major producers of the potent proinflammatory cytokines interleukin-1 beta and tumor necrosis factor-alpha, and of the antiinflammatory cytokine interleukin-1 receptor antagonist . During the adult respiratory distress syndrome the normally surfactant-coated alveolus becomes flooded with plasma proteins, altering the milieu of alveolar cells such as alveolar macrophages . To understand alveolar macrophage function during the adult respiratory distress syndrome, the individual and combined effects of surfactant and plasma on alveolar macrophage cytokine production was examined . A synthetic surfactant (Exosurf) and a bovine-derived surfactant (Survanta) both inhibited production of interleukin-1 beta, pro-interleukin-1 beta, tumor necrosis factor-alpha, and interleukin-1 receptor antagonist in a dose-dependent manner . This inhibition was noted when both endotoxin and heat-killed Staphylococcus aureus were used as stimuli . Autologous plasma also inhibited interleukin-1 beta and tumor necrosis factor-alpha release in a dose-dependent manner, but, unlike surfactant, plasma did not inhibit interleukin-1 receptor antagonist release . Similarly, the combination of plasma and surfactant inhibited interleukin-1 beta and tumor necrosis factor-alpha release but not interleukin-1 receptor antagonist release . In support of these data, interleukin-1 receptor antagonist was detectable in five of six bronchoalveolar lavage fluid samples from patients with adult respiratory distress syndrome at a mean concentration of 465 pg/ml; on the other hand, interleukin-1 beta was not detectable in any of these samples . These results indicate that the relative production of interleukin-1 beta, tumor necrosis factor-alpha, and interleukin-1 receptor antagonist can be altered depending on the local concentration of both surfactant and plasma. J Med Microbiol, 1995 Mar, 42(3), 214 - 9 Mechanisms of 4-quinolone resistance in quinolone-resistant and methicillin-resistant Staphylococcus aureus isolates from Japan and China; Tanaka M et al.; Ninety-two and 33 methicillin-resistant Staphylococcus aureus (MRSA) strains were isolated in Japan and China respectively . They were categorised as ofloxacin-susceptible (MIC < 12.5 mg/L), moderately (MIC 12.5-25 mg/L) or highly (MIC > or = 50 mg/L) ofloxacin-resistant . 4-Quinolone concentrations required to inhibit purified DNA gyrase from the moderately and highly quinolone-resistant MRSA were at least 20 times higher than those required to inhibit the equivalent enzyme from quinolone-susceptible strains . Reconstitution assays demonstrated that the 4-quinolone-resistant MRSA had a mutation in subunit A of DNA gyrase . A portion of the gyrA gene from amino acids codons 40-115 was sequenced . Four moderately resistant and seven highly resistant MRSA contained a Ser-->Leu substitution at amino acid 84; one moderately and one highly resistant MRSA and one moderately resistant methicillin-susceptible S . aureus (MSSA) strain contained a Glu-->Lys substitution at amino acid 88 . Eight MRSA, including one quinolone-susceptible strain and one MSSA contained a silent mutation at amino acid 86 . Uptake of ofloxacin in moderately resistant strains was almost the same in the presence or absence of carbonyl cyanide m-chlorophenylhydrazone (CCCP), whereas in highly resistant strains, uptake increased when CCCP was added . Restriction fragment length analysis of the norA gene with the restriction endonuclease SfcI showed a mutation of nucleotide position 1085 in all MRSA strains tested except for one highly quinolone-resistant strain . Thus the mechanisms of 4-quinolone-resistance in these MRSA isolates involved alterations in both DNA gyrase and antimicrobial uptake and efflux. J Bacteriol, 1995 Mar, 177(6), 1491 - 6 Identification of endo-beta-N-acetylglucosaminidase and N-acetylmuramyl-L-alanine amidase as cluster-dispersing enzymes in Staphylococcus aureus; Sugai M et al.; Two proteins which are capable of dispersing cell clusters of Staphylococcus aureus have been purified from a S . aureus FDA209P culture supernatant . Both of them were found to have bacteriolytic activity . From the elution profile of column chromatography and Western blot (immunoblot) analysis, one of them was identified as a 51-kDa endo-beta-N-acetylglucosaminidase (GL) . The other was a 62-kDa protein on the basis of sodium dodecyl sulfate gel electrophoresis . Analysis of the peptidoglycan fragments following treatment with the 62-kDa protein indicated that this protein is an N-acetylmuramyl-L-alanine amidase (AM) . In vitro studies of cluster dispersion activities using S . aureus mutant strains Lyt66 or S . aureus Wood46 grown as clusters demonstrated that these two enzymes act synergistically to disperse clusters into single cells . Antiserum against the 51-kDa GL cross-reacted with the 62-kDa AM, and S . aureus FDA209P grown in the presence of anti-51-kDa-GL immunoglobulin G induced giant clusters . Clusters induced by anti-51-kDa GL and by Cibacron blue F3G-A were dispersed by coincubation with the 51-kDa GL and the 62-kDa AM . Western blot analysis demonstrated that the 51-kDa GL and the 62-kDa AM were missing in culture supernatants of S . aureus Lyt66, Wood46, and RUSAL2 (Tn551 autolysin-defective mutant), which grow in clusters . These results strongly suggest that the 51-kDa GL and 62-kDa AM are involved in cell separation of daughter cells after cell division. J Infect Dis, 1995 Mar, 171(3), 614 - 24 Eradication of endemic methicillin-resistant Staphylococcus aureus infections from a neonatal intensive care unit; Haley RW et al.; To control infections with endemic methicillin-resistant Staphylococcus aureus (MRSA) in a neonatal intensive care unit (NICU), triple dye was applied to the umbilical cords of infants in the intermediate-care but not the intensive-care area . The rate of MRSA infection, adjusted for time and intensity of care, decreased in the intermediate-care area (rate ratio, 0.35; 95% confidence interval {CI}, 0.14-0.87; P < .01) but not in the intensive-care area (rate ratio, 0.92; 95% CI, 0.41-2.24; P = .48) . After 22 months, the rate increased in both areas (Mantel-Haenszel rate ratio, 1.7; 95% CI, 1.0-2.8; P < .05) after overcrowding and understaffing increased . After temporary reduction of overcrowding and understaffing, extension of triple dye use to the intensive-care area and dedication of an infection control nurse to the NICU, MRSA colonization and infection rates decreased to near zero in both areas (infection rate ratios, 0.09 and 0.11, respectively; P < .005) . The endemic MRSA strain, identified by pulsed-field gel electrophoresis, was eradicated. Antimicrob Agents Chemother, 1995 Mar, 39(3), 714 - 9 Mode of action of the lantibiotic mersacidin: inhibition of peptidoglycan biosynthesis via a novel mechanism? Brotz H, Bierbaum G, Markus A, Molitor E, Sahl HG. Mersacidin is an antibiotic peptide produced by Bacillus sp . strain HIL Y-85,54728 that belongs to the group of lantibiotics . Its activity in vivo against methicillin-resistant Staphylococcus aureus strains compares with that of the glycopeptide antibiotic vancomycin (S . Chatterjee, D . K . Chatterjee, R . H . Jani, J . Blumbach, B . N . Ganguli, N . Klesel, M . Limbert, and G . Seibert, J . Antibiot . 45:839-845, 1992) . Incubation of Staphylococcus simulans 22 with mersacidin resulted in the cessation of growth and slow lysis . Biosyntheses of DNA, RNA, and protein were not affected, whereas incorporation of glucose and D-alanine was inhibited and a regular reduction in the level of cell wall thickness was observed . Thus, unlike type A lantibiotics, mersacidin does not form pores in the cytoplasmic membrane but rather inhibits cell wall biosynthesis . Comparison with tunicamycin-treated cells indicated that peptidoglycan rather than teichoic acid metabolism is primarily affected . Mersacidin caused the excretion of a putative cell wall precursor into the culture supernatant . The formation of polymeric peptidoglycan was effectively inhibited in an in vitro assay, probably on the level of transglycosylation . In contrast to vancomycin, the activity of mersacidin was not antagonized by the tripeptide diacetyl-L-Lys-D-Ala-D-Ala, indicating that on the molecular level its mode of action differs from those of glycopeptide antibiotics . These data together with electron microscopy suggest that mersacidin acts on a novel target, which opens new perspectives for the treatment of methicillin-resistant S . aureus. Vopr Med Khim, 1995 Mar-Apr, 41(2), 40 - 2 {The effect of Staphylococcus aureus toxic shock exotoxin on whole blood cell chemiluminescence in vitro and in vivo}; Antipov AIu et al.; Stimulated and nonstimulated blood chemiluminescence were studied in presence of the toxic shock syndrome toxin I (TSST-I) in vivo and in vitro . In vivo experiments involved evaluation of rabbit whole blood chemiluminescence developed within various periods after the TSST-I intraperitoneal administration . Effect of various TSST-I concentrations on blood chemiluminescence was studied in vitro . TSST-I was shown to inhibit the nonstimulated chemiluminescence in blood within 45 min after administration of LD50 of both opsonized zymosan and calcium ionophore A 23187 in vivo . Similar effects the toxin exhibited at concentrations 20 microM and more in vitro . The maximum effect TSST-I demonstrated within the first 5 min of incubation and this effect was distinctly dose-dependent. Pediatr Dermatol, 1995 Mar, 12(1), 12 - 5 The frequency of erythromycin-resistant Staphylococcus aureus in impetiginized dermatoses; Misko ML et al.; A trend toward increasing resistance of Staphylococcus aureus to standard antibiotic therapy has been reported . Specimens were taken from 98 patients in our outpatient staff clinic who had clinical signs of superficial skin infections . Patients with erythromycin-resistant S . aureus were contacted by telephone or seen in clinic . The organism was found in 87% of patients . Twenty-two (26%) of the 85 cultures that grew S . aureus were resistant to erythromycin . Treatment failure occurred in one of these patients . We conclude that S . aureus is the most common causative organism in secondary skin infections and impetigo in our patient population . Despite significant erythromycin resistance, there was a low frequency of treatment failure in this group . Erythromycin may still be a reasonable agent in the treatment of uncomplicated superficial skin infections in our community at this time. J Antimicrob Chemother, 1995 Mar, 35(3), 421 - 4 Susceptibility of methicillin-resistant Staphylococcus aureus to the essential oil of Melaleuca alternifolia; Carson CF et al.; All 66 isolates of Staphylococcus aureus tested were susceptible to the essential oil of Melaleuca alternifolia, or tea tree oil, in disc diffusion and modified broth microdilution methods . Of the isolates tested, 64 were methicillin-resistant S . aureus (MRSA) and 33 were mupirocin-resistant . The MIC and MBC for 60 Australian isolates were 0.25% and 0.50%, respectively . Comparable results were obtained by co-workers in Britain using similar methods . These in-vitro results suggest tea tree oil may be useful in the treatment of MRSA carriage. J Antimicrob Chemother, 1995 Mar, 35(3), 399 - 408 A double-blind, randomized, placebo-controlled clinical trial to evaluate the safety and efficacy of mupirocin calcium ointment for eliminating nasal carriage of Staphylococcus aureus among hospital personnel; Fernandez C et al.; Sixty-eight health care workers were enrolled in a double-blind clinical trial and randomized to receive either mupirocin calcium ointment or placebo, intranasally bid for 5 days . Nasal cultures were taken immediately before starting treatment, 1 and 2 during treatment, at the end of treatment, 3 days later, weekly for 1-5 weeks and then monthly for 2-6 months after treatment . Mupirocin eliminated nasal carriage with Staphylococcus aureus in 58% of subjects within two days and 86.7% subjects by the end of therapy compared to 9.4% subjects at the end of treatment with placebo (P < 0.001) . Post-treatment colonization rates of 43%, 56% and 67% were attained after 1 month, 2-4 and 6 months treatment with mupirocin respectively and recolonisation with the same strain of S . aureus that had been isolated before treatment was noted in 32%, 40% and 48% . No resistance to mupirocin developed and the drug was well tolerated . Mupirocin is safe and effective in suppressing nasal carriage of S . aureus. J Am Soc Nephrol, 1995 Mar, 5(9), 1697 - 702 Polymorphonuclear leukocyte oxidative burst is enhanced in patients with chronic renal insufficiency; Ward RA et al.; Previous reports that polymorphonuclear leukocyte (PMN) function is impaired in hemodialysis patients do not differentiate between effects of dialysis and of uremia . The hypothesis that chronic renal insufficiency impairs PMN function was tested . Phagocytosis and oxidative burst were measured in PMN from patients with varying degrees of chronic renal insufficiency impairs PMN function was tested . Phagocytosis and oxidative burst were measured in PMN from patients with varying degrees of chronic renal insufficiency (creatinine clearance, 6 to 35 mL/min per 1.73 m2) and normal subjects . The ability of tumor necrosis factor-alpha (TNF-alpha) to prime the oxidative burst was also assessed . Phagocytosis of Staphylococcus aureus and basal H2O2 and O2- release by PMN did not differ between normal subjects and patients with chronic renal insufficiency . However, the oxidative burst stimulated by S . aureus and formyl-Met-Leu-Phe, but not phorbol myristate acetate, was significantly enhanced in PMN from patients with chronic renal insufficiency . The increase in formyl-Met-Leu-Phe-stimulated oxidative burst correlated significantly with the level of renal function . TNF-alpha significantly increased S . aureus-induced H2O2 production in normal PMN, but not in PMN from patients with chronic renal insufficiency . These data indicate that chronic renal insufficiency does not impair PMN phagocytosis and oxidative burst . To the contrary, it enhances receptor-mediated oxidative burst . The inability of TNF-alpha to further enhance the oxidative burst suggests that PMN exist in a primed state in patients with chronic renal insufficiency. J Burn Care Rehabil, 1995 Mar-Apr, 16(2 Pt 1), 97 - 103 Cytotoxicity testing of topical antimicrobial agents on human keratinocytes and fibroblasts for cultured skin grafts; Boyce ST et al.; Cultured epidermal skin has become an adjunctive therapy for treatment of major burn injuries, but its effectiveness is greatly limited because of destruction by microbial contamination . To evaluate candidate antimicrobial agents for use with cultured skin, a combined cytotoxicity-antimicrobial assay system was developed for determination of toxicity to cultured human keratinocytes and fibroblasts and for determination of susceptibility or resistance of common burn wound organisms . Candidate agents including chlorhexidine gluconate, polymyxin B, mupirocin, sparfloxacin, or nitrofurazone were tested separately for inhibition of growth of human cells and for inhibitory activity to microorganisms with the wet disk assay . The data showed that (1) chlorhexidine gluconate (0.05%) was uniformly toxic to both cultured human cells and microorganisms; (2) nitrofurazone (0.02%) had dose-dependent toxicity to human cells and limited effectiveness against gram-negative microorganisms; (3) sparfloxacin (30 micrograms/ml) had low toxicity to human cells and retained antimicrobial activity against both gram-positive and gram-negative bacteria; (4) polymyxin B (400 U/ml) was not toxic to human cells and had intermediate effectiveness on gram-negative bacteria; and (5) mupirocin (48 micrograms/ml) had no toxicity to skin cells and had uniform effectiveness against Staphylococcus aureus including methicillin-resistant Staphylococcus aureus . Selection of topical antimicrobial drugs by these assays may improve effectiveness of cultured skin for burns and may be used to control other surgical wound infections. Am J Orthop, 1995 Mar, 24(3), 262 - 4 A septic hip complicated by a ruptured appendix in an intravenous drug user; Pereles TR et al.; This paper reports on a case of hip joint sepsis complicated by a ruptured appendix in an intravenous drug user . A 41-year-old woman underwent open irrigation and debridement of her right hip joint for a methicillin-sensitive Staphylococcus aureus infection . Five days later the patient developed an intraperitoneal mass, requiring laparotomy and debridement of a periappendiceal abscess . The organisms infecting the abscess were different from those infecting the patient's hip . The patient recovered satisfactorily after 6 weeks of intravenous antibiotic therapy. Infect Dis Clin North Am, 1995 Mar, 9(1), 11 - 24 Staphylococcus aureus infections in diabetic patients; Breen JD et al.; Staphylococcus aureus infections may occur with greater frequency among patients with diabetes mellitus . This article reviews the available literature as it pertains to diabetes and S . aureus in three categories: colonization/carriage, bacteremia with or without metastatic complications, and dialysis-related infections . The clinical entity of pyomyositis is also discussed. Clin Chest Med, 1995 Mar, 16(1), 111 - 20 Pneumonia due to Staphylococcus aureus infection; al-Ujayli B et al.; Staphylococcus aureus is the second most common infectious agent of pneumonia in the ICU . The virulence of this organism is highlighted by toxins and enzymes that result in severe damage to lung tissue . Clinical features fail to distinguish Staphylococcus aureus pneumonias from other pathogens, and clinical diagnosis has the same limitations that beset other bacterial causes of pneumonia . Effective therapy is dictated by carefully performed susceptibility testing . First-line therapy is with a beta-lactam agent . If BRSA is detected or beta-lactam intolerance occurs, vancomycin should be administered . Despite agents active in vitro, the mortality of this disease remains high, especially if spread through hematogenous routes. QJM, 1995 Mar, 88(3), 181 - 9 Prognostic significance of the neutrophil count in immunocompetent patients with bacteraemia; Leibovici L et al.; To examine the prevalence of neutropaenia in immunocompetent, bacteraemic patients, and whether it carries an independent risk for mortality, we surveyed 2096 bacteraemic patients without malignant diseases, and who were not receiving cytotoxic drugs . The granulocyte count on the day of the first positive blood culture was < 1 x 10(9) cells/l in 33 patients (1.7%, group 1); 1.0-4.0 x 10(9) cells/l in 154 patients (7.9%, group 2); 4.0-8.0 x 10(9) cells/l in 564 patients (29%, group 3); 8.0-20.0 x 10(9) cells/l in 1034 patients (53%, group 4); and > 20.0 x 10(9) cells/l in 163 patients (8.4%, group 5) . The mortality rates in the five groups were 39.4%, 18.8%, 18.1%, 25.7% and 25.8%, respectively (p = 0.0001) . The main pathogens in group 1 were Staphylococcus aureus in 25% of patients and Pseudomonas sp . in 23% . Mortality in group 1 patients was higher than in the other patients (odds ratio 1.4, 95% CI 1.1-1.9} . Mortality was also significantly higher in group 2 patients with high blood urea nitrogen . The percentage of neutropaenia, septic patients without known risk factors for neutropaenia is small, but their mortality is high . Overall mortality in patients with relative neutropaenia (1.0-4.0 x 10(9) cells/l) is low, but a subgroup of patients with high blood urea nitrogen is at considerable risk for a fatal outcome . High leucocyte counts are also a marker of increased risk for mortality, but this association is not an independent prognostic factor. Heart Lung, 1995 Mar-Apr, 24(2), 177 - 8 Escherichia coli sternal osteomyelitis after open heart surgery; Shea KW et al.; Deep-seated infections after open-heart surgical procedures, fortunately, are uncommon with appropriate prophylactic antibiotics and careful aseptic technique . When serious infection, such as sternal osteomyelitis, does occur, the effects are devastating and usually require one or more debridement procedures . The organisms usually implanted in postoperative sternal infections are primarily Staphylococcus aureus and aquatically based gram-negative bacilli . Common gram-negative pathogens such as Escherichia coli are very unusual in this setting . We report a case of E . coli sternal osteomyelitis in a diabetic patient after coronary artery bypass grafting. Chemotherapy, 1995 Mar-Apr, 41(2), 77 - 81 Changes of surface hydrophobicity and charge of Staphylococcus aureus treated with sub-MIC of antibiotics and their effects on the chemiluminescence response of phagocytic cells; Nomura S et al.; The effects of the sub-MIC of antibiotics on the surface hydrophobicity and charge of Staphylococcus aureus were examined by the contact angle method and by microscopic electrophoresis, and the production of oxygen-derived radicals by mouse peritoneal macrophages was measured by a luminol-chemiluminescence assay . The treatment of the bacterial cells with antibiotics induced an increase in hydrophobicity and a decrease in the negative charge of the bacterial surface . The chemiluminescence of the macrophages stimulated by S . aureus treated with antibiotics was significantly higher than that obtained with the untreated bacterial cells . These findings suggest that the antibiotics caused an increase in the hydrophobicity and a decrease in the negative charge of the surface of S . aureus, resulting in the enhancement of nonopsonic phagocytosis of S . aureus by macrophages. Jpn J Antibiot, 1995 Mar, 48(3), 402 - 8 Synergistic enhancement of in vitro antimicrobial activity of imipenem and cefazolin, cephalothin, cefotiam, cefamandole or cefoperazone in combination against methicillin-sensitive and -resistant Staphylococcus aureus; Uete T et al.; Synergistic enhancement of the in vitro antimicrobial activity of imipenem combined with cephalosporins against methicillin-resistant Staphylococcus aureus (MRSA) has been reported . In order to investigate which cephalosporin is more effective in enhancing the activity of imipenem against MRSA, the in vitro antimicrobial activities of imipenem, cefazolin, cephalothin, cefotiam, cefamandole and cefoperazone, alone and in combination, against methicillin-sensitive Staphylococcus aureus (MSSA) and MRSA were assessed . Using the checkerboard Mueller-Hinton agar dilution method, strong synergy was found in 97% to 100% of MRSA strains for imipenem and all tested cephalosporins except cefoperazone; fractional inhibitory concentration (FIC) indices were < or = 0.5 . Among the cephalosporins studied, cefamandole most markedly increased the activity of imipenem against MRSA, followed, in order of decreasing effect, by cefotiam, cephalothin, cefazolin, and cefoperazone . The synergistic effect of imipenem combined with cefamandole or cefotiam was confirmed using the broth dilution method with 2% of NaCl. J Clin Microbiol, 1995 Mar, 33(3), 551 - 5 Pulsed-field gel electrophoresis as a replacement for bacteriophage typing of Staphylococcus aureus; Bannerman TL et al.; Bacteriophage typing (BT) (World Health Organization method) has been used at the Centers for Disease Control and Prevention for over 30 years to type isolates of Staphylococcus aureus . Since studies have shown that BT patterns have poor reproducibility and because BT fails to type a high percentage (15 to 20%) of isolates, the Centers for Disease Control and Prevention has converted from using BT to using pulsed-field gel electrophoresis (PFGE) for strain typing S . aureus . We compared the results of BT with results of PFGE for typing 300 isolates of S . aureus, including strains from several well-characterized outbreaks . Ninety-six isolates were BT group I, 19 were group II, 82 were group III, 7 were group V, and 96 were nontypeable . PFGE identified subgroups within each phage group and thus was more discriminating than BT, which identified no subgroups . PFGE was able to type all isolates and distinguish related from unrelated strains of S . aureus . Our modified, standardized PFGE methodology should enable typing laboratories to obtain rapid, reliable results in 3 to 4 days when starting with an isolated colony on agar media. J Pediatr Orthop, 1995 Mar-Apr, 15(2), 169 - 71 Radial-nerve palsy associated with septic shoulder in neonates; Lejman T et al.; Four neonates presented with septic shoulders after delays in diagnosis of 7-20 days . All lacked active extension of the wrist and thumb, and three lacked active extension of the fingers . Two had positive cultures for Staphylococcus aureus, and one had a positive Escherichia coli culture . Surgical treatment was one single aspiration and three arthrotomies . Recovery of extension began in 10-21 days and was normal at 18-35 days post-treatment . At follow-up 9-13 months later, all limbs were neurologically normal . Cadaver dissections showed that the radial nerve passed close to the shoulder joint, and when the capsule was distended, it impinged on the nerve. Cornea, 1995 Mar, 14(2), 175 - 9 Effect of topical antioxidant therapy on experimental infectious keratitis; Alio JL et al.; To test the effect that the treatment with topical antioxidants may have on corneal infection, we have studied the effect of topically applied antioxidants, such as dimethylthiourea 0.5% (DMTU) and dismutase superoxide 0.2% (SOD), on infectious experimental keratitis caused by Staphylococcus aureus . We have quantified the results of the incubated corneas in ex vivo as well as in in vivo treated with antioxidants by using the luminol amplified chemiluminescence technique (LAC) . The evaluation of corneal inflammation was performed calculating the average inflammatory index obtained from the clinical observation of the corneal secretion, corneal edema and ciliary injection . The evolution of the corneal infiltration was evaluated by means of computerized planymetry . The antioxidants used in this study demonstrated a significant reduction of the LAC values when compared with a control group both in the in vivo as well as in ex vivo studies . No significant differences in the clinical evaluation of the average inflammatory index were observed between the study and the control groups . However, a significant increase in the corneal infiltration was registered in the antioxidant treated group (p < 0.001) evaluated by computerized planymetry . Our results indicate that the use of antioxidants as antiinflammatory drugs may have a potential negative influence on the course of infectious keratitis. J Dermatol, 1995 Mar, 22(3), 175 - 80 Staphylococcus aureus isolated from nostril anteriors and subungual spaces of the hand: comparative study of medical staff, patients, and normal controls; Namura S et al.; An epidemiologic investigation of methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus aureus (S . aureus) colonization was conducted at Kansai Medical University Hospital between 1990 and 1991 . The incidence of nasal and subungual positivity for S . aureus was examined in a total of 156 subjects including inpatients, physicians, and nurses at a ward for dermatology, plastic surgery, and emergency patients, outpatients with atopic dermatitis and other skin diseases, and normal controls . Inpatients were most heavily colonized with MRSA (40.8%), but S . aureus colonization was most frequent in outpatients with atopic dermatitis (95.5%) . Not only nostrils, which have been much discussed as a reservoir of S . aureus, but also subungual spaces seemed to be havens of S . aureus . Twelve out of 22 atopic dermatitis patients were positive for S . aureus on skin regions, and coagulase and phage testing showed a correlation between the nasal and skin-colonizing S . aureus . Coagulase type II and phase type NT (not typable) were the predominant types of S . aureus, including MRSA. J Antibiot (Tokyo), 1995 Mar, 48(3), 233 - 42 Total structures and antimicrobial activity of bacitracin minor components; Ikai Y et al.; Total structures of 13 minor components of bacitracin (BC) were proposed, and their antimicrobial activities were investigated . The components of BC including bacitracins A (BC-A) and F (BC-F) were isolated by preparative HPLC and were hydrolyzed under acidic conditions . The resulting amino aids were derivatized with 1-fluoro-2,4-dinitrophenyl-5-L-alanineamide and were separated by HPLC to determine their absolute configurations . It was found that the N-terminal amino acids of BC-A and its related components were epimerized during the hydrolysis to yield their enantiomers . The formation of these artifactual amino acids suggests that our previously proposed structures of the BC minor components are incorrect; therefore, the structures were corrected based on these results . The structures of the BC minor components were the same as that of BCs-A and -F except that one to three of the L-isoleucines, including the N-terminal one, were replaced by L-valines . These structures were confirmed by tandem mass spectrometry under fast atom bombardment (FAB) conditions and Frit-FAB liquid chromatography/mass spectrometry . Based on the UV spectra of the BC components determined by photodiode array detection-HPLC analysis, a new systematic nonmenclature was proposed for the minor components . The isolated components were also used for the determination of their minimal inhibition concentrations and it was found that BC-A is 2 approximately 8 times more potent than the other minor components against strains of Micrococcus luteus and Staphylococcus aureus. FEMS Microbiol Lett, 1995 Mar 1, 126(3), 305 - 9 Nucleotide and deduced amino acid sequence of the gene for a novel protein with a possible regulatory function encoded in the beta operon of Staphylococcus aureus; Aboshkiwa MA et al.; Although considerable homology exists between the translation products of the rplL, rpoB and rpoC genes of the beta operons of the Gram-negative organism Escherichia coli and the Gram-positive Staphylococcus aureus the region between the rplL and rpoB genes is quite different in the two bacterial species . In E . coli the 324 bp has three centres of dyad symmetry in the first half of the sequence and multiple nonsense codons in all three reading frames . By contrast, the corresponding region in S . aureus consists of 1000 bp capable of forming a similar arrangement of stem-loop structures but with an open reading frame, sited 177 bp downstream of the end of rplL and 217 bp upstream of the beginning of the rpoB gene, with consensus initiation and termination signals, which if translated would generate a 22,665 Da protein with 202 amino acids . In view of the inability to find any significant homology with other proteins in the data bank and because the evidence suggests, as in E . coli, that the rplL-rpoB intergenic sequence is involved in regulation it is proposed that the expression product of orf202 may be a further element of control in the S . aureus beta operon. J Orthop Res, 1995 Mar, 13(2), 286 - 95 Contaminated fractures of the tibia: a comparison of treatment modalities in an animal model; Curtis MJ et al.; External fixation is the current standard treatment for skeletal stabilization of open tibial fractures, but intramedullary fixation techniques have become increasingly popular . The aim of this study was to compare, in an animal model, the susceptibility to infection of contaminated fractures stabilized with external fixation with that of contaminated fractures fixed with intramedullary locking nails with or without reaming . A unilateral osteotomy of the tibia was performed in 15 goats under general anesthesia . Each osteotomy was stabilized with either (a) a unilateral biplanar external fixator, (b) an 8 mm diameter intramedullary rod inserted without reaming of the medullary cavity, or (c) a 10 mm diameter rod inserted after reaming . A standardized inoculum of Staphylococcus aureus, 10(3) colony forming units per milliliter, was placed at each osteotomy site on a piece of absorbable gelatin sponge, to simulate contamination of an open fracture . Antibiotics were not administered . The animals were allowed full activity after the procedure . Fourteen days postoperatively, the animals were killed, radiographs of the tibiae were taken, and the tibiae were harvested in a sterile manner . Multiple specimens for quantitative microbiological analysis were taken from the fracture site and from sites 3 cm distal and 6 cm proximal to the fracture . Additional specimens of bone were taken for histological study . Clinical, radiographic, and microbiological analysis demonstrated that, in this animal model, there were significantly fewer and less severe infections in fractures fixed with external fixation than in those fixed with an intramedullary nail with or without reaming . There was marked cortical necrosis in tibiae that had been fixed with nailing and reaming. Clin Diagn Lab Immunol, 1995 Mar, 2(2), 166 - 71 Effects of culture conditions on production of type 5 capsular polysaccharide by human and bovine Staphylococcus aureus strains; Poutrel B et al.; Two Staphylococcus aureus strains, the prototype human Reynolds strain and a bovine isolate, were grown in different complex media and in a synthetic medium (D . Taylor and K . T . Holland, J . Appl . Bacteriol . 66:319-329, 1989) and compared for their ability to produce type 5 capsular polysaccharide . Cell-bound and cell-free type 5 capsular polysaccharide were measured by a new one-step competition enzyme-linked immunosorbent assay . The total production and the proportion of cell-bound type 5 capsular polysaccharide were dependent on the nature of the medium, the duration of the culture, and the strain . Both strains produced more type 5 capsular polysaccharide when cultivated in the synthetic medium than when cultivated in complex media . The best yield of type 5 capsular polysaccharide, about 300 micrograms/ml of medium, was obtained with strain Reynolds grown for 48 h with shaking in the synthetic broth containing glucose as a carbon source. Biomaterials, 1995 Mar, 16(5), 361 - 7 In vitro biocompatibility of a polyurethane catheter after deposition of fluorinated film; Pizzoferrato A et al.; The in vitro biocompatibility of an experimental surface-treated polyurethane was compared with an untreated polyurethane already used for intravascular catheters . The experimental surface was coated with a fluorinated film using a glow discharge treatment . Neither of the catheters was cytotoxic for L929 murine fibroblasts, caused platelet adhesion or release reaction, or changed the mean platelet volume . The surface-treated polyurethane, however, caused a higher adhesion of Staphylococcus aureus than did the untreated one . Therefore, using in vitro testing, it has been ascertained that the examined material, though not being cytotoxic and not modifying platelet behaviour, could favour bacterial adherence. Zentralbl Hyg Umweltmed, 1995 Mar, 196(6), 545 - 61 {Microbial resistance to formaldehyde . III> Dependence of the microbial effect on Staphylococcus aureus, Enterococcus faecium and spores of Bacillus stearothermophilus on temperature}; Spicher G et al.; Temperature dependence of microbicidal efficacy of formaldehyde was examined with suspension tests (pH 7.0) . Test germs were Staphylococcus aureus, Enterococcus faecium and spores of Bacillus stearothermophilus . The methodology was nearly the same as in previous investigations (5, 7) . At given exposure periods and temperatures formaldehyde concentrations necessary to produce a microbicidal effect of log (N/N0) = -4.0 (concentrations of equal efficacy) were determined . N and N0 represent the numbers of colony-forming units in suspensions with and without formaldehyde, respectively . On rectangular graphic representation with the reciprocal value of absolute temperature on the abscissa and with the logarithm of the formaldehyde concentration on the ordinate, the formaldehyde concentrations of equal efficacy fitted straight lines . Lines referring to different exposure periods nearly paralleled each other . With increasing exposure periods the steepness of the lines decreased slightly . This effect was most pronounced with Staphylococcus aureus as a test germ . The ratio of formaldehyde concentrations of equal efficacy for exposure periods of 120 minutes at 20 degrees C and 30 degrees C, respectively, was 3.1:1 with Staphylococcus aureus, and 2.8:1 with Enterococcus faecium . The corresponding ratio obtained with spores of Bacillus stearothermophilus and referring to 60 degrees C and 70 degrees C, respectively, was 3.6:1 . The logarithms of these ratios decreased with temperature in the same measure as the pertinent absolute temperatures increased . On the basis of the previously presented three-dimensional model of the relations between concentration, period of action and efficacy of microbicidal agents, it could be shown that deviations of the results from a linear and parallel course reflect an inconstant concentration exponent . When low formaldehyde concentration, long exposure period and "high" temperature coincide, the efficacy of formaldehyde is lower than calculated for a linear and parallel course of the relation. J Hosp Infect, 1995 Mar, 29(3), 177 - 88 Methicillin-resistant Staphylococcus aureus in Western Australia, 1983-1992; Riley TV et al.; A statewide screening programme has prevented imported strains of methicillin-resistant Staphylococcus aureus (MRSA) from becoming established in any hospital in Western Australia (WA) . Recently, notifications of MRSA in WA have increased, prompting a review of surveillance data for the period 1983-1992 . Our aims were to determine: (i) the distribution by age and sex of persons with MRSA; (ii) changes in notification rates over time and by location in WA; and (iii) temporal changes in antimicrobial resistance patterns . There were 631 notifications of MRSA for the 10 year period 1983-1992, ranging from a low of 36 in 1988 to a high of 117 in 1992 . When the distribution by age and sex was examined, three age group peaks were apparent: 0-9 years, 20-39 years and 60-79 years . There was a predominance of females in the 20-39 years age group, reflecting a greater proportion of hospital nursing staff carrying MRSA . In those aged 50 years or more, there was a marked predominance of males . The highest notification rates overall occurred in the remote Kimberley region of WA, however, rates increased significantly in all regions of the state in 1992 . Based on antimicrobial resistance patterns, MRSA was classified into two groups: multiresistant imported strains which often caused outbreaks in hospitals; and a less resistant MRSA (WA MRSA) . WA MRSA appears to have originated in the Kimberley region and then spread widely in the community to other regions of the state, and the proportion of WA MRSA has increased significantly since 1989. Eur J Clin Microbiol Infect Dis, 1995 Mar, 14(3), 199 - 205 Diversity of methicillin-resistant Staphylococcus aureus isolated in a Canadian hospital; Hammerberg O et al.; Three neonates and three other patients located elsewhere in the hospital became infected with Staphylococcus aureus . Initial automated microdilution susceptibility testing with oxacillin and disk diffusion testing with amoxicillin-clavulanic acid indicated the isolates had borderline oxacillin resistance (MICs 4 micrograms/ml), presumably due to hyperproduction of beta-lactamase . Chromosomal DNA restriction fingerprinting and phage typing revealed the neonatal isolates to be identical; whereas, the other patients were infected with three different strains . Further analysis of the four strains by Southern hybridization with a mecA specific oligoprobe and a quantitative beta-lactamase assay demonstrated that two strains carried the mecA gene (coding for low affinity penicillin-binding protein 2a), and two strains were hyperproducers of beta-lactamase, including one which was mecA gene positive . One strain neither carried the mecA gene nor hyperproduced beta-lactamase . The two mecA gene positive strains displayed oxacillin MICs of 16 micrograms/ml on dilution susceptibility testing in 4% NaCl supplemented Mueller-Hinton agar . Hence, they were considered intrinsically methicillin-resistant Staphylococcus aureus . Both oxacillin and amoxicillin-clavulanic acid MICs were increased on NaCl supplementation . Results of amoxicillin-clavulanic acid disk diffusion susceptibility testing did not correlate with quantitative beta-lactamase production . It is recommended that clinical laboratories do not use amoxicillin-clavulanic acid disk diffusion assays to differentiate suspected borderline resistance due to beta-lactamase hyperproduction from mecA gene expression of PBP-2a since additional mechanisms may account for resistance. Int J Urol, 1995 Mar, 2(1), 17 - 23 Combined intraarterial cisplatin infusion and radiation therapy for invasive bladder cancer; Mizoguchi H et al.; Combined intraarterial cisplatin infusion and radiation therapy were performed as the initial treatment for 23 patients (mean age: 70 years) with invasive bladder cancers (T2 in 17, T3 in 6) who were suitable for total cystectomy . Of these patients, five who had multiple invasive cancers without laterality had their intrapelvic hemodynamics altered by embolizing a contralateral internal iliac artery . Cisplatin (50 mg) was infused into the internal iliac artery through a subcutaneous reservoir twice a week over three weeks while concurrent radiation therapy with 30 Gy, delivered in 15 fractions, was performed . Additional cisplatin infusions were given in six patients . After this combined therapy, total cystectomy and ileal conduit was performed in six patients and transurethral resection of bladder tumor (TURBT) in 17 . Two of the patients who underwent total cystectomy were found to exhibit a complete response . Therefore, the overall response rate was 87%, including 13 complete responses and seven partial responses . The complete response rates in patients with clinical stage T2 and T3 disease were 53 and 67%, respectively . The complete response rate was slightly higher in patients with a non-papillary cancer than in those with a papillary one . Toxic reactions included a decrease in bladder capacity in two patients and severe diarrhea due to methicillin-resistant Staphylococcus aureus colitis in one . Other forms of toxicity, including nausea, vomiting, neurotoxicity in the gluteal region, nephrotoxicity and myelosuppression, were tolerable . All but one of the patients are alive . This patient died of distant metastasis and seven other patients had a local recurrence of bladder cancer.(ABSTRACT TRUNCATED AT 250 WORDS) Infect Control Hosp Epidemiol, 1995 Mar, 16(3), 170 - 4 An overview of nosocomial infection control in Brazil; Pannuti CS et al.; Brazil is the largest country in Latin America, with a population of 146 million people . The socioeconomic development and the distribution of population and health services varies widely within the country . There are approximately 1.2 million hospital admissions per month, 80% of them paid by a government healthcare program that follows the diagnosis-related groups (DRGs) model . The Ministry of Health has been trying to establish a nationwide nosocomial infection control program since 1983 . Most Brazilian hospitals now have some kind of infection control activity, but only a few of them have complete programs . Infrastructural deficiencies, the scarcity of well-trained healthcare workers, and the widespread occurrence of multiresistant Staphylococcus aureus and gram-negative bacteria are some of the challenges faced by Brazilian hospitals in the control of nosocomial infection. Electrophoresis, 1995 Mar, 16(3), 366 - 76 An improvement of restriction analysis of bacteriophage DNA using capillary electrophoresis in agarose solution; Kleparnik K et al.; Seven representatives of the serogroup B Staphylococcus aureus bacteriophages, 29, 53, 55, 83A, 85, phi 11 and 80 alpha, were examined by capillary electrophoresis (CE) for genomic homology using DNA restriction analysis . Genomic DNA of individual bacteriophages was cleaved by HindIII restriction endonuclease, and the resulting restriction fragments were separated by standard horizontal agarose slab gel electrophoresis (SGE) as well as by CE in low-melting-point agarose solutions . The number and size of restriction fragments identified by both methods were compared . The high separation power of CE makes it possible to extend the restriction fragment patterns . In most of the restriction patterns, some additional restriction fragments as small as 150 bp, not identified by SGE, were detected . With respect to speed, high separation efficiency, low sample consumption and automation, CE offers a simple procedure for processing of multiple samples cost-effectively in a reasonable time . The comparison of the complemented restriction patterns of the different phage strains and the subsequent identification of their common fragments leads to a deeper understanding of their phylogenetic relationships . The genome homologies expressed for individual phage pairs in terms of coefficient F values ranged from 15 to 69% . These values are in good accordance with the degree of DNA homology of these phages as determined by DNA hybridization studies and thermal denaturation analysis of DNA by other authors . The total size of each phage genome was estimated by adding the sizes of individual restriction fragments. Vet Immunol Immunopathol, 1995 Mar, 45(1-2), 31 - 43 Monocyte function in cattle experimentally infected with bovine immunodeficiency-like virus; Rovid AH et al.; The effects of bovine immunodeficiency-like virus (BIV) on monocyte function were examined in experimentally infected cattle and in monocytes infected in vitro . Infection with the R29 isolate of BIV appeared to have relatively little effect on monocyte function in cattle during the first 2 years postinfection (PI) . For the first 4 to 8 months post infection, monocyte phagocytosis of Staphylococcus aureus tended to be lower (P = 0.06) in BIV infected calves than in control animals . After 8 months PI, however, phagocytosis became equal between the two groups . Random and chemotactic migration and antibody-dependent cell-mediated cytotoxicity (ADCC) did not appear to be affected by BIV infection . Monocytes from BIV infected cattle were able to respond to in vitro treatment with interferon gamma similarly to monocytes from control cattle . Although experimental infection with BIV R29 resulted in minimal effects on monocyte function, this result could have been due either to a low virus burden in vivo or because BIV is intrinsically unable to affect monocyte function . To distinguish between these possibilities, monocytes from control, uninfected cattle were treated with BIV virus in vitro . Treatment of normal monocytes with cell-free virus significantly (P < 0.05) increased phagocytosis and random and chemotactic migration and decreased ADCC, in a dose-dependent manner . It appears, therefore, that the normal function of peripheral blood monocytes in the BIV R29 infected animals may be due to a low virus burden rather than to the inability of BIV to affect monocyte function . The in vitro infection results also raise the possibility that the function of monocyte derived cells at local sites of BIV replication may be altered. Immunol Lett, 1995 Mar, 45(3), 210 - 4 Differential regulation of surface immunoglobulin expression by various muramyl dipeptides in a murine pre-B cell line; Cohen LY et al.; The murine pre-B cell line 70Z/3 responds to lipopolysaccharide (LPS), interleukin-1 (IL-1) or interferon-gamma (IGN gamma) by kappa gene transcription and expression of surface IgM (sIg) . We found that muramyl dipeptide (MDP), a synthetic immunoadjuvant analog of a bacterial membrane structure, produced a weak increase in the number of sIg-positive 70Z/3 cells as measured by immunofluorescence staining . This number was significantly increased after exposure to MDP . Moreover, when MDP was used in combination with LPS, IL-1 or IFN gamma, an enhancement of sIg expression was observed showing an early influence of MDP in the presence of a second stimulant . Unexpectedly, two adjuvant-active analogs of MDP did not share its capacity to stimulate differentiation of the cell line when used alone or associated with other agents, indicating that adjuvanticity of MDP was not the only requirement . Two other products of bacterial origin, a Staphylococcus aureus cell extract (SAC) and the Toxic Shock Syndrome Toxin TSST-1 could neither enhance the kappa gene expression in 70Z/3 cells nor increase the MDP effect . The stimulating effect displayed by MDP could by related to NF-kappa B activation. Br J Pharmacol, 1995 Mar, 114(6), 1317 - 23 Delayed circulatory failure due to the induction of nitric oxide synthase by lipoteichoic acid from Staphylococcus aureus in anaesthetized rats; De Kimpe SJ et al.; 1 . This study investigates the effect of lipoteichoic acid (LTA) from the cell wall of Staphylococcus aureus, a micro-organism without endotoxin, on haemodynamics and induction of nitric oxide synthase (iNOS) in the anaesthetized rat . 2 . Intravenous injection of LTA (10 mg kg-1) resulted in a decrease in blood pressure from 123 +/- 1 mmHg to 83 +/- 7 mmHg after 270 min (P < 0.001) and a reduction of the pressor response to noradrenaline (1 microgram kg-1) from 33 +/- 1 mmHg.min to 23 +/- 3 mmHg.min after 270 min (P < 0.05) . 3 . The delayed circulatory failure (hypotension and vascular hyporeactivity) caused by LTA was prevented by pretreatment of rats with dexamethasone (10 mg kg-1, 60 min prior to LTA) or the nitric oxide synthase inhibitor NG-monomethyl-L-arginine (L-NMMA, 10 mg kg-1 h-1, i.v . infusion starting 30 min prior to LTA) . 4 . In contrast, treatment of rats with polymyxin B (0.05 mg kg-1), an agent which binds endotoxin (lipopolysaccharides, LPS), did not affect the delayed circulatory failure caused by LTA . Polymyxin B, however, attenuated the hypotension and vascular hyporeactivity to noradrenaline afforded by endotoxaemia (2 mg kg-1 LPS, i.v.) for 270 min . 5 . The delayed circulatory failure caused by LTA was associated with a time-dependent increase in (i) the expression of iNOS protein in the lung (Western blot analysis), and (ii) iNOS activity . This increase in iNOS protein and activity was prevented by pretreatment of LTA-rats with dexamethasone (10 mg kg-1).(ABSTRACT TRUNCATED AT 250 WORDS) Immunopharmacology, 1995 Mar, 29(2), 111 - 9 The effect of GM-CSF and G-CSF on human neutrophil function; Bober LA et al.; A direct comparison of GM-CSF and G-CSF in a panel of in vitro neutrophil-function assays was performed to investigate any differences in activity profiles . In our modified chemotactic assay, GM-CSF rapidly increased the migratory capacity of polymorphonuclear cells (PMNs) to move toward fMLP and LTB4 . In contrast, G-CSF only stimulated PMN migration towards fMLP . GM-CSF, but not G-CSF, increased PMN cytotoxic killing of C . albicans blastospores . The expression of PMN surface antigens associated with Fc- and complement-mediated cell-binding (Fc gamma R1, CR-1 and CR-3), and adhesion signalling (ICAM-1), was increased after the exposure of GM-CSF, but not to G-CSF . In contrast these CSFs demonstrated relative equipotency in their ability to induce PMN anti-bacterial phagocytosis, and to restore the Staphylococcus aureus killing capacity of dexamethasone-suppressed neutrophils . The phagocytic activity of PMNs for opsonized yeast, as well as hexose-monophosphate shunt activity, was equivalent following GM-CSF or G-CSF treatment . We discuss the significance of the difference in activity profiles in this article. J Infect Dis, 1995 Mar, 171(3), 607 - 13 Effects of Staphylococcus aureus leukocidins on inflammatory mediator release from human granulocytes; Konig B et al.; The secretion of the Panton-Valentine leukocidin (Luk-PV) but not of another leukocidin (Luk-R) from Staphylococcus aureus strains is correlated with severe pyodermic infections (dermonecrosis) . The effects of both Luk-PV and Luk-R in amounts of 0-5000 ng on inflammatory mediator release from human leukocytes were studied . Luk-PV but not Luk-R induced a pronounced release of the vasodilator histamine from human basophilic granulocytes (up to 55% +/- 7%) and of enzymes (beta-glucuronidase, up to 45% +/- 10%; lysozyme, up to 35% +/- 7%), chemotactic components leukotriene B4 (42 +/- 8 ng/10(7) cells) and interleukin-8 (up to 33 +/- 5 ng/10(7) cells), and oxygen metabolites from human neutrophilic granulocytes . The results indicate that granulocytes play a central role in dermonecrosis; these in vitro data account for the histologic picture of Luk-PV infections, characterized by local vasodilation, infiltration of granulocytes, and a central necrotic area. Infect Immun, 1995 Mar, 63(3), 1095 - 101 Role of a carboxy-terminal site of toxic shock syndrome toxin 1 in eliciting immune responses of human peripheral blood mononuclear cells; Drynda A et al.; Staphylococcus aureus toxic shock syndrome toxin 1 (TSST-1) is involved in the pathogenesis of toxic shock syndrome and perhaps other staphylococcal diseases . Recently, the C-terminal part of the TSST-1 toxin has been shown to be responsible for mitogenic activity in animal models . We studied the role of the C-terminal structural unit of TSST-1 with regard to proliferation, cytokine release (tumor necrosis factor alpha {TNF-alpha}, interleukin-6 {IL-6}, and IL-8), mRNA expression for IL-6, IL-8, IL-10, TNF-alpha, and CD40 ligand (CD40L), synthesis of immunoglobulin E (IgE), IgA, IgG, and IgM, CD23 expression, and soluble CD23 (sCD23) release from human peripheral blood mononuclear cells (PBMC) . For this purpose, we used the recombinant wild-type TSST-1 (p17) mutant toxin Y115A (tyrosine residue modified to alanine) and toxin H135A (histidine residue modified to alanine) . Unmodified toxin p17 and mutant toxin Y115A, at a concentration below 5 ng, to a lesser degree, induced a strong proliferation . Toxin p17 followed by toxin Y115A was the most pronounced inducer for mRNA expression for IL-10 and CD40L and cytokine generation (mRNA and protein) for TNF-alpha, IL-6, and IL-8 . Mutant protein H135A failed to activate human PBMC . Both toxins p17 and, to a lesser degree, Y115A significantly suppressed IL-4- and anti-CD40-induced synthesis of all four Igs as well as IL-4-induced CD23 expression and sCD23 release . Mutant toxin H135A failed to do so . Thus, our data show that a region in the C terminus of TSST-1 is responsible not only for mitogenic activity but also for additional immunomodulating biological activities of TSST-1 . More specifically, histidine residue H135A of the 194-amino-acid toxin appears to be critical for the expression of biological activities in a human in vitro model. Protein Eng, 1995 Mar, 8(3), 315 - 8 Generation of analogs having potent antimicrobial and hemolytic activities with minimal changes from an inactive 16-residue peptide corresponding to the helical region of Staphylococcus aureus delta-toxin; Dhople VM et al.; The delta-toxin is a 26-residue peptide from Staphylococcus aureus with the sequence formyl-MAQDIISTIGDLVKWIIDTVNKFTKK . NMR studies indicate that the segment IISTIGDLVKWIIDTV occurs in an alpha-helical conformation in the toxin . A synthetic peptide corresponding to this segment, although helical, did not exhibit hemolytic activity . Since charged residues like D and K are likely to modulate cytolytic activity, analogs of the 16-residue peptide were synthesized where D was systematically replaced by K . Analogs in which the first D and both Ds were replaced by K showed potent antimicrobial and hemolytic activities . The analog in which the second D was replaced by K was relatively less active . However, all the peptides showed an alpha-helical structure with similar helical content . The activities of the peptides were found to correlate directly with their ability to permeabilize model membranes . Thus, by minimal judicious replacement of charged amino acids, it should be possible to generate cytolytic peptides from short segments of peptide toxins. Proc Natl Acad Sci U S A, 1995 Feb 28, 92(5), 1619 - 23 Autocrine regulation of toxin synthesis by Staphylococcus aureus; Balaban N et al.; Staphylococcus aureus is a major human pathogen causing diseases which range from minor skin infection to endocarditis and toxic shock syndrome . The pathogenesis of S . aureus is due primarily to the production of toxic exoproteins, whose synthesis is controlled by a global regulatory system, agr . We show here that agr is autoinduced by a proteinaceous factor produced and secreted by the bacteria and that it is inhibited by a peptide produced by an exoprotein-deficient S . aureus mutant strain . The inhibitor, RIP, competes with the activator, RAP, and may be a mutational derivative . Our results suggest two possible approaches, independent of antibiotics, to the control of S . aureus infections . RIP may prove useful as a direct inhibitor of virulence and RAP as a vaccine against the expression of agr-induced virulence factors; either could interfere with the ability of the bacteria to establish and maintain an infection. Biochem Biophys Res Commun, 1995 Feb 27, 207(3), 978 - 84 Isolation and the gene cloning of an alkaline shock protein in methicillin resistant Staphylococcus aureus; Kuroda M et al.; The growth of Staphylococcus aureus occurs at a wide range of pH(5-10), while the optimal is pH 7.0-7.5 . The molecular mechanism of such pH tolerant properties should be elucidated because the production of the virulence factors was greatly affected by environmental pH . The effect of pH shift on the composition of cytosolic proteins in S . aureus was examined . A protein with a molecular mass of 23 kDa was remarkably enhanced by a pH upshift from 7 to 10 . This alkaline shock protein (ASP23) was isolated and purified by ion-exchange chromatography . The N-terminal sequences of the purified protein and the protease-digested peptides were analyzed . The 320-bp DNA fragment that was designed from the peptide analysis was amplified . Using the amplified fragment as a probe, the ASP gene, asp23, was cloned . The deduced primary sequence of ASP23 comprised 169 amino acids with a calculated molecular weight of 19,191 . Northern analysis revealed that asp23 was positively regulated at the transcription level by alkaline shock . Homology search revealed that asp23 is a novel gene . Although the physiological role of ASP23 has yet to be further analysed, we suggest that ASP23 plays a key role in alkaline pH tolerance of S . aureus. Arch Biochem Biophys, 1995 Feb 20, 317(1), 85 - 92 Characterization of the glycosaminoglycan-binding region of lactoferrin; Wu HF et al.; Lactoferrin is a prominent component of neutrophil secondary granules and its blood concentration is increased in certain inflammatory diseases . Although the biochemical characterization of lactoferrin as an iron-binding protein has been well described, its physiological role in inflammation remains undefined . We examined the ability of lactoferrin to regulate glycosaminoglycan-accelerated thrombin-serine protease inhibitor (serpin) reactions . Lactoferrin effectively reduced the rate of thrombin-serpin (antithrombin and heparin cofactor II) reactions by three physiological glycosamino-glycans including heparin, heparan sulfate, and dermatan sulfate . An enzyme kinetics analysis showed that lactoferrin did not alter the apparent heparin-thrombin or the heparin-antithrombin dissociation constant values for the heparin-catalyzed thrombin-antithrombin reaction . However, the maximum reaction velocity at saturation with respect to either protein was markedly decreased by lactoferrin . The glycosaminoglycan-binding region of lactoferrin was analyzed following limited proteolysis using Staphylococcus aureus V8 protease . Two lactoferrin fragments with Mr's of approximately 8 and approximately 11 kDa were purified based on their affinity to heparin-Sepharose . Amino acid sequence analysis demonstrated that both peptides were from the N-terminus . Although slightly less capable compared to intact lactoferrin, the lactoferrin peptides effectively neutralized heparin, heparan sulfate, and dermatan sulfate-catalyzed serpin-thrombin inhibition reactions . In addition, lactoferrin N-terminal peptides have approximately the same binding affinity to heparin-Sepharose as that of intact lactoferrin . Inspection of both the N-terminal amino acid sequence and the crystal structure of lactoferrin further supports the conclusion that lactoferrin is a novel glycosaminoglycan binding protein and that the putative glycosaminoglycan-binding site is localized to the N-terminus. Med Clin (Barc), 1995 Feb 18, 104(6), 221 - 3 {The pediatric heart transplant . Its evolution and early postoperative complications}; Villaizan C et al.; BACKGROUND: Cardiac transplantation is an acceptable therapeutic alternative for cardiac diseases refractory to other forms of management in adults as well as in infants and children . METHODS: Between 1987-1992 7 children (4 girls and 3 boys) underwent cardiac transplantation: four with dilated cardiomyopathy, one with cardiac fibroma and two with hypertrophic cardiomyopathy . Age at transplantation ranged from 2 months to 13 years and 5 months, with a follow-up ranging from 15 months to 5 years and 9 months . Prophylaxis of acute rejection consisted of cyclosporine, azathioprine and glucocorticoids . RESULTS: Two patients presented acute rejection three weeks after cardiac transplantation, with a good response to high dose glucocorticoids . Two patients developed severe infection (sepsis by Staphylococcus aureus) with successful outcome after antibiotic treatment . One patient died in the early postoperative period and other after 4 years 11 months postransplantation because myelodysplastic syndrome . At present only one case is receiving glucocorticoids in immunoprophylaxis . The status is asymptomatic in the other 5 patients with a normal height-weight development . CONCLUSIONS: Heart transplantation provides durable therapy for congenital and myopathic heart disease in infants and children with an excellent quality of life. J Biol Chem, 1995 Feb 17, 270(7), 3365 - 9 The N-terminal portion of growth inhibitory factor is sufficient for biological activity; Uchida Y et al.; To determine its active site, growth inhibitory factor (GIF), a central nervous system-specific metallothionein-like protein, was digested with trypsin followed by Staphylococcus aureus protease V8 digestion . Of 5 peptide fragments separated from trypsin-digested GIF by reverse-phase high pressure liquid chromatography and gel filtration, only GIF1-26 or longer peptides showed growth inhibitory activity on cortical neurons in culture . A shorter peptide, GIF5-23, which was obtained by further digestion of GIF1-26 with V8 protease, also showed growth inhibitory activity . However, a synthetic peptide corresponding to GIF5-23 did not show growth inhibitory activity . Metal-free GIF1-26 prepared by acid treatment showed a similar level of growth inhibitory activity to that of metal-containing GIF1-26, indicating that metal in the peptide does not affect the activity . Treatment of metal-free GIF1-26 with beta-mercaptoethanol resulted in the loss of activity . The CD spectrum of beta-mercaptoethanol-treated metal-free GIF1-26 was different from that of nontreated metal-free GIF1-26 . These results indicate that the N-terminal portion of GIF is required for growth inhibitory activity and that folding of the peptide via S-metal bonding is critical for biological activity. J Immunol, 1995 Feb 15, 154(4), 1606 - 13 Role of IL-12 in human B lymphocyte proliferation and differentiation; Jelinek DF et al.; The role of IL-12 in human peripheral blood B cell responsiveness was examined . To analyze the ability of IL-12 to directly mediate B cell growth and/or differentiation, FACS-purified (> 99% pure) B cells were studied and a polyclonal B cell-activating system utilizing Cowan I Staphylococcus aureus was used . Whereas IL-2 is highly effective in this system in promoting both B cell growth and differentiation, IL-12 was observed only to augment modestly B cell growth and to be ineffective by itself as a B cell differentiation factor for S . aureus-stimulated B cells . However, IL-12 markedly enhanced Ig secretion when added in the presence of IL-2 . Moreover, when the ability of IL-12 to augment IL-2-dependent B cell Ig secretion was compared with the ability of several known auxiliary B cell differentiation factors, IL-12 was observed to be the most potent cytokine that could costimulate with IL-2 . Analysis of IL-12-stimulated B cell cultures failed to reveal outgrowth of T cells and NK cells . In addition, assessment of IFN-gamma levels in IL-12-driven B cell culture supernatants and analysis of IFN-gamma effects on B cell responses added additional support to the conclusion that IL-12 directly modulates B cell function . Finally, these results suggest that IL-12 is a potent constimulus of B cell differentiation and that the signals conveyed by IL-12 seem to be qualitatively distinct from the differentiative signals delivered by other cytokines such as IL-2. FEMS Microbiol Lett, 1995 Feb 15, 126(2), 139 - 43 The CAMP effect of Actinobacillus pleuropneumoniae is caused by Apx toxins; Jansen R et al.; Actinobacillus pleuropneumoniae shows synergistic haemolysis when cocultured with Staphylococcus aureus on blood agar plates . This CAMP effect has been attributed to a discrete CAMP factor, but also to the A . pleuropneumoniae-RTX-toxins I, II, and III . We examined the CAMP effect of recombinant Escherichia coli strains that secreted each of these toxins, and of A . pleuropneumoniae mutant strains that were devoid of one or more these toxins . We found that the E . coli strains were CAMP positive, whereas the A . pleuropneumoniae strain devoid of functional toxin genes was CAMP negative . This demonstrated that the CAMP effect of A . pleuropneumoniae is caused by the toxins and that no CAMP factor per se exists. Biochemistry, 1995 Feb 14, 34(6), 1988 - 96 Purification and characterization of two potent heat-stable protein inhibitors of protein phosphatase 2A from bovine kidney; Li M et al.; Two heat-stable protein inhibitors of protein phosphatase 2A (PP2A), tentatively designated I1PP2A and I2PP2A, have been purified to apparent homogeneity from extracts of bovine kidney . The purified preparations of I1PP2A exhibited an apparent M(r) approximately 30,000 and 250,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel permeation chromatography on Sephacryl S-300, respectively . In contrast, the purified preparations of I2PP2A exhibited an apparent M(r) approximately 20,000 and 80,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel permeation chromatography on Sephacryl S-200, respectively . The purified preparations of I1PP2A and I2PP2A inhibited PP2A with 32P-labeled myelin basic protein, 32P-labeled histone H1, 32P-labeled pyruvate dehydrogenase complex, 32P-labeled phosphorylase, and protamine kinase as substrates . By contrast, I1PP2A and I2PP2A exhibited little effect, if any, on the activity of PP2A with 32P-labeled casein, and did not prevent the autodephosphorylation of PP2A in incubations with the autophosphorylation-activated protein kinase {Guo, H., & Damuni, Z . (1993) Proc . Natl . Acad . Sci . U.S.A . 90, 2500-2504} . The purified preparations of I1PP2A and I2PP2A had little effect, if any, on the activities of protein phosphatase 1, protein phosphatase 2B, protein phosphatase 2C, and pyruvate dehydrogenase phosphatase . With 32P-labeled MBP as a substrate, kinetic analysis according to Henderson showed that I1PP2A and I2PP2A were noncompetitive and displayed a Ki of about 30 and 25 nM, respectively . Following cleavage with Staphylococcus aureus V8 protease, I1PP2A and I2PP2A displayed distinct peptide patterns, indicating that these inhibitor proteins are the products of distinct genes . The N-terminal amino acid sequences of the purified preparations indicate that I1PP2A and I2PP2A are novel proteins. Anal Biochem, 1995 Feb 10, 225(1), 64 - 72 A microtiter plate assay using cascade amplification for detection of nonisotopically labeled DNA; Rothschild CB et al.; We describe a microtiter-plate-based, colorimetric assay for DNA, the enzyme-linked DNA-enzyme-linked coagulation assay (EDNA-ELCA) . The EDNA-ELCA uses amplification of the common pathway of coagulation for the ultrasensitive detection of DNA which is tagged by incorporation of functional groups such as biotin and fluorescein . The EDNA-ELCA enables detection of attomole amounts of DNA (< 1 pg per microtiter well), with a sensitivity 200-1000 times higher than other colorimetric techniques . The assay has been applied as an adjunct to PCR for quantitative determination of methicillin-resistant Staphylococcus aureus DNA at levels corresponding to 1-10(5) organisms . The EDNA-ELCA can also be used to assay DNA by hybridization; < 50 amol of an unlabeled DNA template is detected by hybridization to biotin- and fluorescein-labeled probes. Kyobu Geka, 1995 Feb, 48(2), 156 - 9 {Intermittent mini-dose vancomycin intravenous administration and closed continuous irrigation technique applied to mediastinitis caused by MRSA following mitral valve replacement}; Suetsugu F et al.; Mediastinitis caused by MRSA (Methicillin-Resistant Staphylococcus aureus) remains an intractable infection producing high mortality even in these days of advanced chemotherapy . The authors report a case of mediastinitis due to MRSA complicated with acute renal failure following mitral valve replacement . The patient's mediastinum had been thoroughly cleaned with physiological saline solution with 0.2% povidone iodine, and underwent a chemotherapy regimen of mini-dose vancomycin . The patient made favorable progress and recovered completely . Our patient's progress confirmed that when chemotherapy using vancomycin is administered in a patient whose condition is complicated with acute renal failure, closely monitoring the vancomycin serum concentration is essential . Intermittent mini-dose intravenous administration is sufficient to maintain an effective vancomycin serum concentration . In our case, vancomycin serum concentration measured before and at completion of dialysis revealed no appreciable decline. FEMS Microbiol Lett, 1995 Feb 1, 126(1), 91 - 2 Antibacterial activity of 7-aminocholesterol, a new sterol; Dherbomez M et al.; A new sterol, 7-aminocholesterol, which inhibits growth of Saccharomyces cerevisiae, also displayed antibiotic activity against Gram-positive bacteria . The 50% growth inhibitory concentration against strains of Listeria innocua, L . monocytogenes, Staphylococcus aureus, Enterococcus hirae and Bacillus cereus was 3 microM. J Med Microbiol, 1995 Feb, 42(2), 91 - 5 Superantigenic exotoxin production by isolates of Staphylococcus aureus from the Kawasaki syndrome patients and age-matched control children; Todome Y et al.; Nineteen strains of Staphylococcus aureus were isolated from the throat or the tooth surfaces of 19 cases amongst 127 patients with Kawasaki syndrome (KS) during the acute phases and 11 S . aureus isolates were obtained from five of 17 diseased controls and six healthy controls . The production of exotoxins, particularly superantigenic toxic shock syndrome toxin-1 (TSST-1), coagulase serotype, pigment production, haemolytic activity and tryptophan auxotrophy of these isolates were compared . Among 10 KS S . aureus strains isolated in 1990-1991, five (50%) secreted TSST-1, a higher frequency than two (18%) of 11 control isolates . In contrast, none of the nine KS strains collected in 1984 produced TSST-1 . Four of five TSST-1-secreting KS strains produced white or white to golden pigmentation, whereas the two control strains capable of TSST-1 production formed golden colonies . There were no noticeable differences between S . aureus strains from KS patients and control children in the production of staphylococcal exotoxins A-E, coagulase serotype, haemolysis of sheep erythrocytes and tryptophan auxotrophy . The pathological or aetiological role of a new TSST-1-secreting S . aureus clone in patients with KS was not confirmed. J Med Microbiol, 1995 Feb, 42(2), 127 - 32 Typing of Staphylococcus aureus colonising human nasal carriers by pulsed-field gel electrophoresis; Hu L et al.; Colonisation by Staphylococcus aureus in the nares of 120 outpatients and 63 healthy adults was studied for c . 2 years . Two states of carriage of S . aureus were confirmed: persistent carriage and persistent non-carriage . The states of carriage and non-carriage were quite stable and > 60% of the population of any of the study groups were stable non-carriers . The results of typing the strains isolated from the same individuals at different times with DNA fingerprinting by digestion with SmaI enzyme showed that all the stable carriers were persistently infected with the same strain and that changes in the strain seldom occurred. Epidemiol Infect, 1995 Feb, 114(1), 143 - 51 A comparative study of food retail premises by means of visual inspection and microbiological quality of food; Powell SC et al.; The relationship between visual inspection ratings given to ten food retail premises and the microbiological quality of food samples was examined . Viable counts of bacteria and of Staphylococcus aureus were determined for cooked meat samples from each of the premises . There was no correlation between potential risk of foodborne infection, as assessed by total inspection rating, and bacteriological counts in food (P < 0.05) . Neither was there a consistent relationship between scores given to any component of the total rating and the bacteriological quality of food . The effectiveness of the current UK inspection scheme in assessing risk of foodborne infection is questioned . Inclusion of appropriately weighted criteria such as food temperature abuse is suggested to improve the scheme. Eur J Biochem, 1995 Feb 1, 227(3), 873 - 9 Duodenase, a new serine protease of unusual specificity from bovine duodenal mucosa . Primary structure of the enzyme; Zamolodchikova TS et al.; The complete amino acid sequence of duodenase, a new serine endopeptidase from bovine duodenal mucosa, has been determined . The sequence was reconstructed by the automated sequence analysis of the peptides obtained after cleavage with trypsin, Staphylococcus aureus V8 protease, cyanogen bromide and duodenase . The enzyme is composed of 226 amino acid residues yielding a molecular mass of 29.06 kDa . The presence of six cysteine residues and one potential sugar-chain-binding site at Asn50 was revealed . A predicted catalytic triade characteristic of the serine proteases was traced in the duodenase primary structure at the corresponding positions (His44, Asp87 and Ser181 in the sequence) . Comparison of the sequence of duodenase with the other known primary structures of mammalian serine proteinases reveales the duodenase identity to granzymes from human and mice, human cathepsin G and mast cell chymases from rat, and gives an overall sequence identity of 47-55% with the mentioned enzymes . Alignment of the known serine protease and duodenase primary structures showed unique amino acid residues within the duodenase substrate-binding pocket at positions 189 (Asn) and 226 (Asp) (the bovine chymotrypsinogen A numbering) . These results are discussed with respect to the relation between the duodenase unique residues within the primary specificity pocket S1 and the unusual dual specificity of the enzyme. Can J Cardiol, 1995 Feb, 11(2), 123 - 8 Molecular analysis provides evidence for human to human transmission of Staphylococcus aureus endocarditis in non-drug abusers; Ignaszewski AP et al.; Two cases of invasive Staphylococcus aureus are reported in which human to human transmission resulted in primary bacteremia and endocarditis . The identity of the organism was confirmed by phage typing, antibiograms, coagulase gene polymorphisms and ribotyping . This is the first documented case of such transmission not involving an intravenous drug abuser. Acta Orthop Scand, 1995 Feb, 66(1), 69 - 72 Collagen with gentamicin for prophylaxis of postoperative infection . Staphylococcus aureus osteomyelitis studied in rabbits; Riegels-Nielsen P et al.; In 34 rabbits, both tibiae were inoculated with Staphylococcus aureus . 14 legs received no treatment and served as controls . In 12 legs, the wound was treated with pure collagen and in 18 legs, collagen with gentamicin (Gentacoll) in a dose of 10 mg/kg body weight was applied to the wound before closure . Postoperatively 12 received 10 mg/kg body weight gentamicin intravenously and no local treatment . The animals were killed 7 days after inoculation and evaluated macroscopically and microbiologically for infection . 6 rabbits (12 legs) were used for pharmacokinetic studies only and they were killed after 2, 4, and 18 hours, respectively . 11/14 untreated legs developed a macroscopically acute osteomyelitis . No infection was found in the 18 legs treated with Gentacoll and 1/12 treated with gentamicin systemically had growth of the inoculated bacteria in tissue biopsies . The concentrations of gentamicin in the serum as well as locally reached peak values were well above the MIC value in all groups, with a maximum after 1-2 hours . No gentamicin could be detected after 18 hours, independently of the mode of administration. J Leukoc Biol, 1995 Feb, 57(2), 235 - 41 Retention of phagocytic functions in cryopreserved human monocytes; Hansen JB et al.; Phagocytosis and respiratory burst activity were measured by flow cytometry in fresh and cryopreserved human monocytes, after ingestion of Escherichia coli and Staphylococcus aureus . Mononuclear leukocytes, isolated from 15 healthy donors, were divided into two portions, of which one was examined immediately and the other was cryopreserved for 3 weeks . Morphological characteristics and expression of receptors involved in phagocytosis were similar in fresh and cryopreserved monocytes . Furthermore, both internalization of bacteria and respiratory burst activity remained unchanged after cryopreservation . Transmission electron microscopy confirmed actual internalization of bacteria and not merely bacterial attachment to monocytes . Monocytes were demonstrated to retain integral cellular functions during cryopreservation . This may imply that the method has potential for use in basal and clinical trials. J Lab Clin Med, 1995 Feb, 125(2), 200 - 11 Molecular correlation between in vitro and in vivo activity of beta-lactam and beta-lactamase inhibitor combinations against methicillin-resistant Staphylococcus aureus; Fasola EL et al.; Beta-Lactam resistance in Staphylococcus aureus is associated with beta-lactamase production, with the presence of a new penicillin binding protein (PBP) called PBP2a, with reduced affinity for beta-lactam antibiotics, and with modifications of normal PBPs . We have studied these mechanisms of resistance, in vivo and in vitro, for several beta-lactam antibiotics against both beta-lactamase-producing and non-producing methicillin-resistant S . aureus organisms (MRSA) . Our results showed that all tested agents inhibited binding of labeled penicillin G to many PBPs . The combination of cefoperazone and sulbactam was the best combination, and it inhibited radiolabeled penicillin G binding to PBP2a at a lower concentration than that needed for cefoperazone alone . In vivo, the regimen of cefoperazone plus sulbactam was also more effective than cefoperazone alone . For beta-lactamase-negative strains this correlated with an increased binding affinity of cefoperazone plus sulbactam to PBP2a and PBP4 . The improved efficacy of cefoperazone plus sulbactam versus cefoperazone with a beta-lactamase producing strain was closely related to cefoperazone hydrolysis by beta-lactamase that was inhibited by sulbactam . This study demonstrates that there is more than one effect of beta-lactamase inhibitors when they are combined with beta-lactam antimicrobial agents, and also that there may be a role for these agents in therapy for MRSA infections. J Bone Joint Surg Am, 1995 Feb, 77(2), 258 - 65 Evaluation of treatment modalities for septic arthritis with histological grading and analysis of levels of uronic acid, neutral protease, and interleukin-1; Nord KD et al.; We compared the effectiveness of antibiotics alone and in combination with arthroscopy, arthroscopy with debridement, arthrotomy, or needle aspiration for the treatment of septic arthritis . Each modality has its proponents, but, to our knowledge, no comparative studies have been conducted in animals . We used biochemical and histological analysis to compare these methods of treatment in an experimental model . The right hind knee of thirty goats was injected with 1 x 10(5) Staphylococcus aureus bacilli . The left hind knee was not inoculated and served as the normal control . Seventy-two hours after inoculation, a two-week course of treatment with intramuscular administration of cefuroxime sodium, either alone or in combination with another mode of treatment, was initiated in each of five groups . The cartilage was evaluated histologically with biochemical, enzymatic, and interleukin-1 analyses . Despite the early therapeutic intervention, on the average, there was a 25 per cent loss of uronic acid (t test, p < 0.001) and a 43 per cent increase in neutral protease activity (signed-rank test, p = 0.003) in the treatment groups . There were no significant intergroup differences with regard to the histochemical-histological rating or the levels of uronic acid, neutral protease, or interleukin-1. J Exp Med, 1995 Feb 1, 181(2), 537 - 46 Stimulatory and inhibitory effects of interleukin (IL)-4 and IL-13 on the production of cytokines by human peripheral blood mononuclear cells: priming for IL-12 and tumor necrosis factor alpha production; D'Andrea A et al.; The production of cytokines in monocytes/macrophages is regulated by several different cytokines that have activating or inhibitory effects . Interleukin (IL)-10, IL-4, IL-13, and transforming growth factor (TGF)-beta are usually considered to be the most important macrophage-deactivating factors, with inhibitory effects on cytokine production . Unlike IL-10 and TGF-beta, which appear to act as downmodulators of many phagocytic cell functions, the mode of action of IL-4 and IL-13 is more complex . Addition of IL-4 and IL-13 to peripheral blood mononuclear cell (PBMC) cultures inhibited production of IL-12, tumor necrosis factor (TNF)-alpha, IL-10, and IL-1 beta induced by lipopolysaccharide (LPS) or Staphylococcus aureus added simultaneously with the cytokines . However, pretreatment of PBMC with IL-4 or IL-13 for > or = 20 h enhanced the production of IL-12 and TNF-alpha in response to LPS or S . aureus several fold in these cells; this IL-4-induced priming for the two cytokines was inhibited by anti-IL-4 neutralizing antibodies . IL-4 priming also enhanced the accumulation of IL-12 and TNF-alpha mRNA induced by LPS and S . aureus . The enhanced accumulation of transcripts for the IL-12 p35 and p40 chains by IL-4 priming was reflected in enhanced secretion of both the IL-12 free p40 chain and the p70 heterodimer . These results suggest an unexpected complexity in the regulatory role of IL-4 and IL-13 in immune responses. Infect Immun, 1995 Feb, 63(2), 585 - 90 Use of adhesion-defective mutants of Staphylococcus aureus to define the role of specific plasma proteins in promoting bacterial adhesion to canine arteriovenous shunts; Vaudaux PE et al.; We used an ex vivo canine arteriovenous shunt model, previously developed to study plasma protein adsorption and thrombogenesis on polymeric biomaterials, to define the role of host proteins in promoting adhesion of Staphylococcus aureus . Either polyethylene or polyvinyl chloride tubings were exposed to canine blood for 5, 15, or 60 min at a flow rate of 300 ml/min and then were flushed in phosphate-buffered saline (PBS), cut into 1.5-cm segments, and stored at -70 degrees C . After thawing, each segment was preincubated in 0.5% albumin in PBS to prevent nonspecific staphylococcal attachment to surfaces that were not exposed to blood . Each segment was then incubated with 4 x 10(6) CFU of {3H}thymidine-labelled S . aureus per ml for 60 min at 37 degrees C in an in vitro adhesion assay . Two site-specific mutants of S . aureus were tested: one specifically defective in adhesion to surface-bound fibronectin (FnAd-def) and the other defective in adhesion to fibrinogen (FgAD-def) {corrected} . Compared with their respective parental strains, the FgAd-def, but not the FnAd-def, mutant of S . aureus showed a strong (> 80%) decrease in attachment to ex vivo tubings . The adhesion of each strain of S . aureus onto polyethylene was consistently more than twofold higher than the adhesion onto polyvinyl chloride segments exposed to flowing blood for 5 or 15 min, but adhesion became similar to that on polyvinyl chloride after 60 min of exposure . In conclusion, the specific adhesion-defective mutants of S . aureus suggested that fibrinogen was the most active adhesion-promoting protein in a short-term blood-material interaction . The experimental approach described in this study should prove useful for screening materials thought to be resistant to protein-mediated staphylococcal adhesion and colonization. Infect Immun, 1995 Feb, 63(2), 509 - 15 A mutation at histidine residue 135 of toxic shock syndrome toxin yields an immunogenic protein with minimal toxicity; Bonventre PF et al.; Structure-function studies have revealed that the region between amino acids 115 and 141 of toxic shock syndrome toxin 1 (TSST-1) constitutes a biologically active domain . A critical residue appears to be histidine 135, since a site-directed mutation that alters the histidine to alanine (H135A) results in a loss of mitogenic activity and an absence of toxicity as measured in a rabbit infection model of toxic shock syndrome . We have characterized the mutant toxin further and report here on its immunogenic activity in rabbits and on the protective ability of mutant-specific antibodies in two animal models of toxin-mediated shock . Antibodies raised in rabbits by immunization with the purified H135A are fully cross-reactive with staphylococcal TSST-1 and wild-type recombinant TSST-1 (rTSST-1) expressed in Escherichia coli . The H135A antibodies neutralized the mitogenic activity for murine splenic T cells equally well as did TSST-1-specific polyclonal and monoclonal antibodies . In addition, the H135A antibodies blocked the production of tumor necrosis factor by spleen cells stimulated with rTSST-1 . The toxicities of rTSST-1 and H135A were compared in D-galactosamine (D-GalNH2)-sensitized MRL-lpr/lpr mice . The nontoxicity of H135A was confirmed in this murine model of superantigen-induced septic shock . No toxicity of H135A was demonstrable at doses of 60 micrograms, while doses of rTSST-1 as low as 2 micrograms caused significant mortality within 24 to 72 h after challenge . Furthermore, subsequent to challenge of mice with H135A, no elevation in the serum levels of interleukin-2 or tumor necrosis factor was measurable . Passive immunization with H135A antibodies also protected MRL-lpr/lpr mice against lethal challenge with rTSST-1 . Finally, rabbits actively immunized with purified H135A did not succumb to infection with a transformed strain of Staphylococcus aureus expressing rTSST-1 . Additional animal studies will be required to confirm the immunizing potential of H135A and the efficacy of H135A antibodies as a neutralizing antitoxin. Infect Immun, 1995 Feb, 63(2), 375 - 80 Antibodies to capsular polysaccharides are not protective against experimental Staphylococcus aureus endocarditis; Nemeth J et al.; The protective efficacy of antibodies to the Staphylococcus aureus capsular polysaccharide was examined in a rat model of catheter-induced endocarditis . Capsular antibodies were induced either by active immunization with killed S . aureus or by passive immunization with hyperimmune rabbit antiserum to S . aureus . Control rats were injected with phosphate-buffered saline or passively immunized with normal rabbit serum or rabbit antiserum to a nonencapsulated strain . Animals with indwelling catheters were challenged intravenously with 5 x 10(4) to 4 x 10(6) CFU of the homologous S . aureus strain (capsular serotype 5 strain Reynolds or serotype 1 strain SA1 mucoid) . Both immunized and control rats developed S . aureus endocarditis . The numbers of S . aureus cells recovered from the blood and aortic valve vegetations of immunized rats were similar to those of control rats, indicating that capsule-specific antibodies were not protective . To determine whether the presence of an indwelling catheter interfered with antibody-mediated protection against S . aureus endocarditis, catheters were removed 2 h after insertion in additional groups of rats . An inoculum of 10(8) CFU of strain Reynolds was needed to provoke endocarditis in rats catheterized for 2 h, compared with 5 x 10(4) CFU for rats with indwelling catheters . Passively transferred capsular antibodies were not protective since both immunized and nonimmunized animals developed endocarditis, and quantitative cultures of blood and valvular vegetations revealed no differences between immunized and control animals . The findings of this study indicate that antibodies to the capsular polysaccharide are not protective in the rat model of experimental S . aureus endocarditis. Int Arch Allergy Immunol, 1995 Feb, 106(2), 124 - 33 Responsiveness of peripheral blood mononuclear cells from normal and atopic donors to microbial superantigens; Konig B et al.; Patients with atopic dermatitis (AD) are frequently colonized with Staphylococcus aureus strains secreting exotoxins such as the staphylococcal enterotoxin B (SEB) and A (SEA) . Nonetheless the role of SEB and SEA in AD is yet unknown . We analyzed the responsiveness of peripheral blood mononuclear cells (PBMCs) and isolated T cells from donors with AD and from normal donors to SEB and SEA . PBMCs as well as T cells from normal donors showed a significantly enhanced proliferation after stimulation with enterotoxin B, whereas the 3H-thymidine uptake of the T lymphocytes from patients with AD was markedly suppressed . Furthermore, we show that IFN-gamma mRNA and protein and mRNA for both chains of IL-12 (p35 and p40) are produced in human PBMCs from normal donors upon stimulation with SEB and SEA . In contrast to normal donors T cells from donors with AD predominantly express mRNA for IL-4, IL-5, and only diminished levels for IFN-gamma and IL-12 upon stimulation with SEB and SEA . Furthermore, in contrast to normal donors, PBMCs from donors with AD spontaneously produce high levels of IgE and express increased levels of CD23, the low-affinity receptor for IgE . Nonetheless, the superantigens by themselves, from 0.1 fg up to 1 microgram/10(6) cells, induced neither IgE secretion nor CD23 expression on PBMCs . Moreover, the addition of superantigens to IL-4-treated PBMC cultures diminished or totally suppressed the IL-4-induced IgE synthesis and CD23 expression . No differences were observed between PBMCs from normal donors of donors with AD . Both PBMCs isolated from normal and atopic donors produced high levels of soluble IL-4-receptor (up to 210 +/- 90 pg/ml) . Addition of soluble IL-4-receptor to PBMC cultures downregulated the IL-4-induced IgE synthesis and CD23 expression in unstimulated as well as in SEB-stimulated PBMCs from normal donors and donors with AD . Our results suggest that superantigen-producing staphylococcal strains on the skin of patients with AD may modulate and/or amplify allergic inflammation. Mol Microbiol, 1995 Feb, 15(4), 679 - 87 Localization of the start sites of lagging-strand replication of rolling-circle plasmids from gram-positive bacteria; Dempsey LA et al.; A number of small, multicopy plasmids from Gram-positive bacteria replicate by an asymmetric rolling-circle mechanism . Previous studies with several of these plasmids have identified a palindromic sequence, SSOA, that acts as the single-strand origin (SSO) for the replication of the lagging-strand DNA . Although not all the SSOA sequences share DNA sequence homology, they are structurally very similar . We have used an in vitro system to study the lagging-strand replication of several plasmids from Gram-positive bacteria using the SSOA sequences of pT181, pE194 and pSN2 as representative of three different groups of