J Antimicrob Chemother, 1996 Nov, 38(5), 859 - 63 Effect of paclitaxel alone or in combination on the intracellular penetration and activity of quinolones in human neutrophils; Garcia I et al.; We evaluated the effect of paclitaxel alone or in combination with cisplatin or doxorubicin, on the intracellular penetration and activity of ciprofloxacin, ofloxacin, levofloxacin and sparfloxacin in human polymorphonuclear leukocytes (PMN) . In general, paclitaxel either alone or in combination did not affect the intracellular concentrations of the quinolones evaluated (intracellular to extracellular concentration ratio (C/E > 4) . However, at high concentrations (20 mg/L) paclitaxel decreased the penetration of ofloxacin and levofloxacin although the C/E ratios remained higher than 3 . Paclitaxel alone or in combination affected neither the production of oxygen radicals by PMN nor the intracellular activity of the quinolones against Staphylococcus aureus. J Am Soc Nephrol, 1996 Nov, 7(11), 2403 - 8 Nasal mupirocin prevents Staphylococcus aureus exit-site infection during peritoneal dialysis . Mupirocin Study Group; Plasma concentrations of endotoxin and antiendotoxin antibodies in patients with multiple injuries: a prospective clinical study; Department of Surgery, University of Ulm, GermanyOBJECTIVE: To investigate the time course of endotoxaemia and its effects on the synthesis of antiendotoxin antibodies in patients with multiple injuries . DESIGN: Prospective clinical study . SETTING: University hospital, Germany . PATIENTS: 40 Patients with multiple injuries and 20 healthy volunteers who served as controls . INTERVENTIONS: Blood samples were collected up to 10 days after injury and the concentrations of endotoxin, antiendotoxin antibodies to four endotoxins, and for anti-alpha-haemolysin of Staphylococcus aureus were measured . The kinetics of endotoxaemia and antiendotoxin antibodies were investigated . RESULTS: Endotoxaemia peaked 0-3 hours after injury at 0.425 EU/ml and decreased thereafter to 0.04 EU/ml within five days . Total concentrations of IgM, IgA, and IgG increased continuously after day 3 (p < 0.05), but the specific IgM response to all endotoxins was only temporary and the relative content of specific antibodies to all endotoxins peaked at day 3 (p < 0.05) . Antiendotoxin antibodies of IgM class cross-reacted among themselves . There was no general increase in specific antiendotoxin antibodies of IgA and IgG class . The relative content of specific antibodies to alpha-haemolysin of S aureus of all classes of immunoglobulins (IgM, IgA, IgG) remained on the same level from day 0-10 . CONCLUSION: Multiple injuries are associated with early and temporary endotoxaemia which causes specific increases in antiendotoxin antibodies of the IgM-class . IgM antibodies to endotoxins cross-react among themselves. Proc Assoc Am Physicians, 1996 Nov, 108(6), 455 - 66 Interaction of tumor necrosis factor-alpha and granulocyte colony-stimulating factor on neutrophil apoptosis, receptor expression, and bactericidal function; Sullivan GW et al.; Infected patients are likely to have increased levels of tumor necrosis factor-alpha (TNF-alpha) and may be treated with recombinant human granulocyte colony-stimulating factor (G-CSF) . Recombinant human TNF-alpha activates polymorphonuclear neutrophil (PMN) inflammatory activity . We examined the effect of exposure to TNF-alpha and G-CSF alone and in combination on PMN apoptosis, receptor expression, phagocytosis, and bactericidal function . The results were compared to those obtained with a promoter of PMN apoptosis, cycloheximide . After 24 hr, 27% of PMNs were nonapoptotic, and TNF-alpha (1 unit/ml) showed no change . Cycloheximide (10 micrograms/ml) decreased the number of nonapoptotic cells to 10% of the initial PMN . In contrast, G-CSF (30 ng/ml) decreased apoptosis (57% nonapoptotic PMN after 24 hr) . Both G-CSF and TNF-alpha (but not cycloheximide) induced preservation of PMN Fc gamma RIII (467% and 167% of 24-hr controls, respectively) and beta 2-integrin expression (150% and 168% of 24-hr controls, respectively) . G-CSF (but not TNF-alpha or cycloheximide) stimulated expression of Fc gamma RI (191% of 24-hr control) and Fc gamma RII (267% of 24-hr control) . G-CSF (but not TNF-alpha) maintained the ability of PMN to ingest and kill opsonized Staphylococcus aureus . TNF-alpha decreased the effect of G-CSF on apoptosis, expression of Fc gamma RIII and Fc gamma RI, and bactericidal function . Thus, TNF-alpha promoted expression of Fc gamma RIII and beta 2-integrin receptors, which are important for phagocytic activity, and G-CSF diminished apoptosis, increased Fc gamma receptor expression, and maintained bactericidal function . TNF-alpha counteracted some effects of G-CSF . Interactions of these cytokines in vivo serve to regulate the PMN inflammatory response and bactericidal capacity. Arzneimittelforschung, 1996 Nov, 46(11), 1045 - 53 Stereo-selective calcium antagonistic and binding properties of the enantiomers of lemildipine in vascular tissue of pigs and dogs; Nakayama K et al.; Stereo-selective calcium antagonistic properties of the enantiomers of lemildipine (CAS 94739-29-4, NB-818) were assessed in vascular tissues, including pig coronary artery and dog cerebral artery . Ca2+ antagonistic action of (-)-lemildipine was about 5 times and 100 times more potent than nifedipine and (+)-lemildipine, respectively . (-), (+), and (+/-)-lemildipine showed a slow onset and long duration of Ca2+ antagonistic action . Pretreatment with (-)-lemildipine and (+/-)-lemildipine for 30 min inhibited the contractions produced by 5-hydroxytryptamine and endothelin-1 with pD2 values between 6.0-8.5, whereas (+)-lemildipine and nifedipine were less potent . Of the lemildipine enantiomers, (+)-lemildipine at a low concentration (1 nmol/l) had a slight but significant Ca2+ agonistic action . However, (+)-lemildipine had no apparent effect on the pCa-tension relationship in the dog basilar artery permeabilized with Staphylococcus aureus alpha-toxin . Enantiomers of lemildipine as well as nifedipine competitively antagonized the specific binding of (+)-{3H}PN 200-110 (isradipine) to the membranes of pig coronary artery in the following order: (-)-lemildipine > nifedipine > (+/-)-lemildipine > (+)-lemildipine . These results suggest that the stereo-selective Ca2+ antagonistic actions of lemildipine enantiomers and nifedipine assessed in the functional experiments were well correlated with their potencies for the competition with the (+)-{3H}PN 200-110 binding to the pig coronary artery. FEMS Immunol Med Microbiol, 1996 Nov, 16(1), 51 - 9 Developmental and environmental factors that enhance binding of Bordetella pertussis to human epithelial cells in relation to sudden infant death syndrome (SIDS); Saadi AT et al.; Asymptomatic infection due to Bordetella pertussis has been suggested to be one cause of sudden infant death syndrome (SIDS) . We examined developmental and environmental factors previously found to affect binding of another toxigenic species, Staphylococcus aureus, to human epithelial cells: expression of the Lewis(a) antigen; infection with respiratory syncytial virus (RSV); exposure to cigarette smoke; and the inhibitory effect of breast milk on bacterial binding . Binding of two strains of B . pertussis (8002 and 250825) to buccal epithelial cells was significantly reduced by treating the cells with monoclonal antibodies to Lewis(a) (P < 0.05) and Lewis(x) (P < 0.01) antigens . Both strains bound in significantly greater numbers to cells from smokers compared with cells from non-smokers (P < 0.05) . HEp-2 cells infected with RSV subtypes A or B had higher binding indices for both 8002 (P < 0.001) and 250825 (P < 0.01) . On RSV-infected cells, there was significantly enhanced binding of monoclonal antibodies to Lewis(x) (P < 0.05), CD14 (P < 0.001) and CD18 (P < 0.01); and pre-treatment of cells with anti-CD14 or CD18 also significantly reduced binding of both strains of B . pertussis . Pre-treatment of the bacteria with human milk significantly reduced their binding to epithelial cells . The results are discussed in relation to our three-year survey of bacterial carriage among 253 healthy infants, their mothers and local SIDS cases between 1993-1995 and in relation to the change to an earlier immunisation schedule for infants and the recent decline in SIDS in Britain. Rinsho Byori, 1996 Nov, 44(11), 1093 - 9 {Mini review: a factor enhanced the antibacterial effect of beta-lactams on methicillin-resistant Staphylococcus aureus}; Tajima Y; We found a factor (Factor T) in aged mixtures of tungstate and phosphate which greatly enhances the antibacterial effects of beta-lactams on methicillin-resistant Staphylococcus aureus (MRSA), while Factor T alone did not strongly inhibit bacterial growth . The effect on methicillin-susceptible S.aureus was weak . There was no synergism of Factor T with other classes of antibiotics, nor with other groups of bacteria . Factor T is probably a complex of phosphate and tungstate, and reduced the expression of penicillin-binding protein-2', thus sensitizing the MRSA strains to beta-lactams. Biochem Mol Biol Int, 1996 Nov, 40(4), 779 - 85 Purification and enzymatic peptide mapping of protein synthesis elongation factor-2 from mink and chicken livers; Riis B; This investigation has shown it it possible to purify elongation factor-2 from livers of two rather distinct animal species, minks and chicken, to high homogeneity by employing the same purification procedure . It is also shown that making peptide maps of the factor by the use of Staphylococcus aureus Endoprotease Glu-C gives the same pattern . Combined, these two experimental approaches show that the evolution has conserved this elongation factor in two species with a rather different physiology and choice of feeding substances . It is also shown that using mink or chicken livers as staring material for purification of eEF-2 is both technically and economically sound when obtaining large amounts of highly purified eEF-2 is the ultimate goal. Eur J Biochem, 1996 Nov 1, 241(3), 765 - 71 Phosphorylation and dephosphorylation in the proline-rich C-terminal domain of microtubule-associated protein 2; Sanchez C et al.; The C-terminal domain of microtubule-associated protein 2 (MAP2) contains a proline-rich region and the tubulin-binding domain . We have generated antibodies to follow the phosphorylation state of the proline-rich domain . One of these antibodies (no . 305) has been raised against a synthetic peptide P (sequence RTPGTPGTPSY) phosphorylated at the threonine residues . This sequence is present in the proline-rich region of MAP2 and is phosphorylated in vitro by at least three different proline-directed protein kinases: p42mpk, p34cdc2, and GSK3 (glycogen-synthase kinase 3) alpha/beta . The MAP2 sites phosphorylated by these kinases are different, although all of them phosphorylate the C-terminal domain of MAP2 as determined by Staphylococcus aureus V8 protease mapping . Nonphosphorylated peptide P can be phosphorylated in vitro by all three kinases studied with similar efficiency . In high-molecular-mass MAP2, this sequence is highly phosphorylated in vivo at the late stages of rat development . This motif can be rapidly dephosphorylated in vitro by protein-phosphatase 1 (PP1) and 2A (PP2A) catalytic subunits but not by PP2B. Res Vet Sci, 1996 Nov, 61(3), 231 - 3 Influence of age of the donor sheep on the phagocytosis of Staphylococcus aureus subspecies anaerobius and S aureus by neutrophils; Quiteria JA et al.; The age of the sheep donating neutrophils had a marked influence in vitro on their phagocytic ability with respect to Staphylococcus aureus and S aureus subspecies anaerobius . Neutrophils from lambs five to eight weeks old phagocytosed these organisms significantly less efficiently (P < 0.001) than neutrophils from adults two to four years old . However, neutrophils from the young animals phagocytosed S aureus significantly (P < 0.001) better than S aureus subspecies anaerobius (61.5 v 53.8 per cent), whereas there was no significant difference in the ability of the neutrophils from adult sheep to phagocytose S aureus and S aureus subspecies anaerobius (68.2 v 68.2 per cent). Protein Expr Purif, 1996 Nov, 8(3), 332 - 40 A Trypanosoma cruzi polyantigen obtained by gene fusion: its expression in Staphylococcus aureus and rapid purification; Moreno JI; In order to simplify the large-scale production of three different Trypanosoma cruzi antigens with significant diagnosis value, their coding DNA fragments were fused to produce a single molecule . This tripartite protein was expressed using a shuttle Staphylococcal protein A (SPA) gene fusion vector . The resulting fusion protein location was intracellular when synthesized in Escherichia coli but was also proteolytically degraded during its purification . When the same construct was expressed using the Staphylococcus aureus secretion system, a nondegraded expression product was obtained from the culture medium . A "size effect" seemed to take place in the final yield . The SPA tripartite antigen fusion protein was purified in one step using IgG-Sepharose affinity chromatography . The SPA affinity tail was removed by specific proteolysis with enterokinase and further chromatography on IgG Sepharose . Specific antibodies against individual antigens reacted equally well with the purified tripartite antigen . These results suggest that the simultaneous production of several antigens in a single molecule using the S . aureus secretion system could be a good alternative, when a mixture of cloned antigens is necessary for a strict diagnosis or for immunization experiments. J Dairy Res, 1996 Nov, 63(4), 543 - 53 Cell cycle regulation of mammary epithelial cell detachment by Staphylococcus aureus; Zavizion B et al.; The effect of Staphylococcus aureus on detachment of bovine mammary epithelial cells in culture was examined . Mammary epithelial cells became detached from fresh monolayers following a 3 h incubation in the presence of Staph . aureus M60 . Two different procedures indicated that cell detachment coincided with the S-phase of the cell cycle . The roles of proteinases, toxins and Ca availability in inducing cell detachment were examined . Addition of the proteinase inhibitor phenylmethylsulphonyl fluoride (1 mM) to the culture medium prevented cell detachment . Addition of a combination of purified staphylococcal proteinases XVI and XVII-B to the culture medium of mammary epithelial cells induced cell detachment in the absence of Staph . aureus . Cell detachment may be caused by a staphylococcal proteinase . However, addition of Ca (10 mM) to the culture medium abolished Staph . aureus-induced cell detachment, despite the fact that proteinase activity was still apparently present . Isogenic mutants of Staph . aureus M60, expressing either alpha or beta toxins but not both, induced cell detachment, but to a lesser extent than the wild type . Thus, Ca and toxins play some role during cell detachment . Clones established from detached cells that were washed and replated showed the same susceptibility to Staph . aureus-induced cell detachment as the parental cells . This indicated that there is no subclone of mammary epithelial cells more sensitive to this effect. J Hosp Infect, 1996 Nov, 34(3), 205 - 10 When should healthcare workers be screened for methicillin-resistant Staphylococcus aureus? Lessing MP, Jordens JZ, Bowler IC. The role of screening of healthcare workers (HCWs) in the control of methicillin-resistant Staphylococcus aureus (MRSA) is controversial . It is recommended in guidelines by expert groups in both North America and the United Kingdom, although the role of MRSA carriage by HCWs in outbreaks is not clearly defined . The present report describes the spread of a distinct strain of MRSA to patients by a single HCW on three separate occasions over 27 months . The isolates from this HCW and patient contacts were shown to be indistinguishable by antibiogram and repetitive extragenic palindromic polymerase chain reaction (REP/PCR); none were typeable by lytic phage-typing . Throat carriage of the MRSA probably recurred in this HCW, despite attempts to eradicate it on three occasions . Over the same period, nine other small clusters were seen in the Oxford Hospital Group, involving 66 patients and 22 HCWs colonized, or occasionally infected, with a variety of MRSA strains . In none of these instances could HCWs be implicated in the initiation of an outbreak . The advantages of a screening policy include the determination of the full extent of MRSA-colonization and work exclusion; the disadvantages include detection of transient nasal carriage, disruption of staff routine and stigmatization . Screening of HCWs can be a valuable tool in the control of MRSA outbreaks but it should be used selectively . This strategy remains an important part of a control programme. J Hosp Infect, 1996 Nov, 34(3), 197 - 203 Changes in major populations of methicillin-resistant Staphylococcus aureus in Belgium; Wildemauwe C et al.; A total of 102 epidemic methicillin-resistant Staphylococcus aureus (MRSA) isolates collected in 13 Belgian hospitals during two periods (1981-1985 and 1991-1992) were tested for phage-type, for the presence of aminoglycoside-modifying enzymes (AME), and examined by arbitrarily primed polymerase chain reaction (AP-PCR) . All isolates, but five, belonged to a few distinct phage-types of group III . Most isolates expressed a combination of AAC(6')-APH(2") with APH(3')III, and ANT(4',4") or both . Both phage-typing and AME suggested a change in the MRSA population between the two periods but the AP-PCR method revealed only slight differences. Ann Surg, 1996 Nov, 224(5), 665 - 71 Rapid diagnosis of methicillin-resistant Staphylococcus aureus bacteremia by nested polymerase chain reaction; Kitagawa Y et al.; OBJECTIVE: The purpose of this study was to establish a rapid and sensitive diagnostic method for methicillin-resistant Staphylococcus aureus (MRSA) bacteremia in postoperative patients . SUMMARY BACKGROUND DATA: As a result of diffusion and abuse of third-generation cephalosporin antibiotics in the 1980s in Japan, an outbreak of MRSA infection has been posed . In the field of surgery, severe postoperative infections with MRSA such as MRSA bacteremia, which may lead to multiple organ failure, have emerged with a high mortality . METHODS: Thirty-five patients with high fever (above 38.5 C) or watery diarrhea or both within 2 weeks after gastrointestinal major surgery and 6 healthy volunteers were examined . Nested polymerase chain reaction was used to detect mecA and toxic shock syndrome toxin-1 (TSST-1) genes in blood specimens . RESULTS: The mecA and TSST-1 genes were not detected in the blood samples of any of the six healthy volunteers . In all 12 samples from which MRSA colonies were isolated by blood culture, mecA and TSST-1 genes were detected . Although it took at least 48 hours to identify MRSA by the blood culture method, the presence of mecA and TSST-1 genes was determined by nested polymerase chain reaction method within only 3 to 4 hours after blood sampling . CONCLUSIONS: This method, as a sensitive and rapid monitoring system for MRS bacteremia, would be clinically beneficial for prevention of cross infection and for early determination of appropriate treatment for infected patients. Arch Biochem Biophys, 1996 Nov 1, 335(1), 102 - 8 Comparison of the beta-toxins from Staphylococcus aureus and Staphylococcus intermedius; Dziewanowska K et al.; The beta-toxins produced by Staphylococcus aureus and Staphylococcus intermedius were purified to homogeneity from culture supernatants . Although the toxin from S . aureus has been throughly studied, less is known about its unique counterpart from S . intermedius . This is the first reported purification and analysis of the S . intermedius beta-toxin . Both toxins have similar enzymatic properties, belong to the class of neutral sphingomyelinases C, and have a high specificity for sphingomyelin . They also hydrolyze lysophosphatidylcholine at a much slower rate, but have no activity toward phosphatidylcholine, phosphatidylethanolamine, or phosphatidylserine . The kinetic parameters determined for both proteins (apparent Km 1.4 mM, Vmax 100 mmol/min/microg protein) are identical . Despite these similarities, the size and amino acid composition of the two beta-toxins differ . Molecular mass values, determined by electrophoresis and gel filtration, indicate that the both enzymes are single polypeptides . The decrease in sphingomyelinase activity of S . aureus beta-toxin upon pretreatment with dithiothreitol (DTT) indicates the presence of a disulfide bond in the protein . In contrast, DTT has no effect on the enzymatic activity of S . intermedius beta-toxin . This observation is consistent with the absence of detectable cysteine residue in the protein . N-terminal amino acid sequences determined for the first 19 residues of both beta-toxins also differ, only nine of the first 19 residues are identical . Further evidence that the two proteins differ was obtained by immunological analysis which demonstrated crossreactivity but a lack of identity. Antimicrob Agents Chemother, 1996 Nov, 40(11), 2529 - 34 In vivo efficacies of levofloxacin and ciprofloxacin in acute murine hematogenous pyelonephritis induced by methicillin-susceptible and-resistant Staphylococcus aureus strains; Frosco MB et al.; Levofloxacin, the active L-isomer of ofloxacin, has demonstrated strong activity against Staphylococcus aureus both in vitro and in vivo . In a murine model of hematogenous pyelonephritis, the in vivo efficacies of levofloxacin and ciprofloxacin were evaluated against two methicillin-susceptible and two methicillin-resistant S . aureus strains . All four isolates had virtually identical susceptibilities to levofloxacin and ciprofloxacin . Pyelonephritis was induced in carrageenan-primed mice by an intravenous injection of 0.5 ml of 10(7) CFU of methicillin-susceptible S . aureus isolates per ml or 10(8) CFU of methicillin-resistant S . aureus isolates per ml . At 1 h postinfection, the mice were treated orally with levofloxacin or ciprofloxacin once a day or twice a day (total daily dose of 20 to 160 mg/kg of body weight) for 4 days . Mice were euthanized 24 h after the final treatment, and the kidneys were excised and weighed . The kidneys were prepared for histological examination or were homogenized to determine the numbers of CFU per gram of tissue quantitatively . The reduction in the mean log10 number of CFU per gram as a function of total daily dose was recorded . A dose-response analysis showed that levofloxacin was superior to ciprofloxacin for all four isolates at any dose or regimen tested, independent of the methicillin susceptibility of the isolates . By using an inverse prediction technique, the equivalent effective doses of levofloxacin (once a day) were less than those of ciprofloxacin (twice a day) by 5.2 and 3.2 times, respectively, for methicillin-susceptible S . aureus 9039 and 3087 . For methicillin-resistant S . aureus 667 and 2878, the equivalent effective doses of levofloxacin (once a day) were less than those of ciprofloxacin (twice a day) by 4.1 and 6.4 times, respectively . In a separate study, histological examination of all infected, untreated mice showed moderate to marked hematogenous pyelonephritis . Levofloxacin-treated mice (40 mg/kg once a day) showed no evidence of pyelonephritis in the kidneys, whereas the kidneys of mice treated with the same dose of ciprofloxacin showed only a reduction in the severity of the lesions . Treatment with ciprofloxacin (80 mg/kg twice a day) demonstrated a histology comparable to that of treatment with levofloxacin (40 mg/kg once a day) . Levofloxacin (40 mg/kg once a day) reduced the log10 numbers of CFU per gram by 5 log10; however, ciprofloxacin (80 mg/kg twice a day) reduced the numbers of CFU per gram by only 3 log10 . In the present murine model of pyelonephritis, levofloxacin was superior to ciprofloxacin in preventing pyelonephritis and eradicating S . aureus. Clin Immunol Immunopathol, 1996 Nov, 81(2), 175 - 81 Inhibition of human B cell activation by gold compounds; Hirohata S; The mechanism of action of gold compounds, which are effective in the treatment of rheumatoid arthritis (RA), has not been clearly identified . Although one of the characteristic features of RA is chronic stimulation of B cells, the effects of gold compounds on B cells have not been precisely assessed . We therefore examined the effects of gold sodium thiomalate (GST) on human B cells . IgM production was induced from highly purified B cells obtained from healthy donors by stimulation with Staphylococcus aureus Cowan 1 (SA) plus IL-2 . T cell proliferation and IFN-gamma production were induced from highly purified T cells by stimulation with immobilized mAb to CD3 . As little as 0.1 microg/ml (0.25 microM) GST almost completely suppressed B cell IgM production, whereas it did not suppress T cell proliferation or IFN-gamma production . The inhibition of IgM production by GST is not due to its thiomalate, but is most likely due to its gold components, since thiomalate alone did not inhibit IgM production . GST was required at the initiation of cultures to exert optimal suppressive effects on IgM production . Moreover, GST suppressed the expression of IL-2R (CD25) and transferrin receptor (CD71) on SA-stimulated B cells . These results indicate that GST preferentially inhibits the function of B cells at concentrations much lower than those which inhibit the function of T cells by interfering with the initial activation of B cells . The direct inhibitory effects of GST on human B cell activation described here may contribute at least in part to its therapeutic effect in RA. FEMS Microbiol Lett, 1996 Nov 1, 144(2-3), 241 - 7 The combined effects of plasma and hydrogel coating on adhesion of Staphylococcus epidermidis and Staphylococcus aureus to polyurethane catheters; John SF et al.; The adhesion of three Staphylococcus epidermidis and three S aureus clinical isolates, to uncoated and hydrogel-coated polyurethane catheters was tested, following pretreatment of catheters with human plasma . Plasma significantly decreased the adhesion of S . epidermidis strains to uncoated polyurethane catheters, but had no significant effect on the adhesion to hydrogel-coated catheters . The influence of plasma on adhesion of S . aureus strains to catheters was strain dependent . Plasma significantly increased the adhesion of one strain (SA6) to uncoated catheters . For two other strains (SA3 and SA14) plasma produced no clear effect on their adhesion to uncoated catheters; adhesion values for each strain showed either a small but significant increase or a replicate-dependent increase or decrease . However, plasma significantly increased the adhesion of all S . aureus strains to hydrogel-coated polyurethane catheters . Overall, with the exception of one batch culture of S . epidermidis strain SE3 tested, attachment to plasma-treated hydrogel coated catheters was statistically significantly lower, by up to 85%, than attachment to plasma-treated uncoated catheters for both S . epidermidis and S . aureus. J Infect Dis, 1996 Nov, 174(5), 1001 - 9 Penicillin G-induced microbicidal activity of endothelial cells cultured on gelfoam blocks; Zhang B et al.; A body of evidence has surfaced documenting the ability of endothelial cells cultured on monolayers to phagocytose but not kill bacteria . Several years ago, a new three-dimensional endothelial cell culturing model was developed, which simulated the morphology of the endothelium in small vessels and capillaries . Given that endothelial cells may be derived from the same pluripotent stem cells as macrophages, the question of whether endothelial cells might phagocytose and kill bacteria was explored . Endothelial cells grown on Gelfoam blocks exhibited bactericidal activity towards Staphylococcus aureus, reaching maximal killing of > 90% after 2 h . Evidence documents the involvement of bacterial adherence to the plasma membrane of the endothelial cell . This is followed by phagocytosis of S . aureus, leading to intracellular killing . Penicillin G, included in the endothelial cell growth medium, was found to be a critical factor in the bactericidal activity demonstrated by Gelfoam blocks laden with endothelial cells. Endocrinology, 1996 Nov, 137(11), 4671 - 6 Stable and diffusible pools of nucleotides in pancreatic islet cells; Detimary P et al.; Adenine nucleotides are thought to serve as second messengers in the control of beta-cell function by glucose, e.g . by regulating the activity of ATP-dependent K+ channels . However, their localization in different intracellular pools may mask the biologically relevant changes and complicate the interpretation of measurements in whole cells . The plasma membrane of mouse islet cells was selectively permeabilized by the alpha-toxin from Staphylococcus aureus to allow diffusion of cytoplasmic nucleotides . After permeabilization of cells from freshly isolated islets, approximately 68% of ATP, 45% of ADP, and 52% of AMP rapidly diffused out of the cells, whereas the insulin content hardly varied . The nondiffusible pool of nucleotides was stable for at least 90 min at 4 C, which suggests that it is contained in cellular organelles . The size of this nondiffusible pool decreased proportionally to insulin stores when these were lowered by stimulating secretion to different degrees during culture before permeabilization . From these results, it can be calculated that nondiffusible nucleotides are mainly contained in insulin secretory granules, with a small proportion in another, probably mitochondrial, compartment . Approximately 80% GTP and 30% GDP were present in the diffusible pool, and their relative proportions in the granular pool were only about 20% that of adenine nucleotides . Incubation of the cells in 20 instead of 2 mM glucose before permeabilization did not affect the nondiffusible pool, which indicates that the increase in the ATP/ADP ratio measured in intact cells occurred in the diffusible pool . Cytoplasmic nucleotide levels could be evaluated by subtracting the nondiffusible pool from the measurements in intact cells . It emerges that glucose induces large changes in the ATP/ADP ratio in the cytoplasmic pool, and that these changes are largely due to a fall in ADP. J Immunol, 1996 Nov 1, 157(9), 4133 - 40 Escherichia coli hemolysin and Staphylococcus aureas alpha-toxin potently induce neutrophil adhesion to cultured human endothelial cells; Krull M et al.; Adhesion of polymorphonuclear leukocytes (PMN) to endothelial cells is an essential step in inflammatory reactions . We characterized the effects of two important bacterial exotoxins, Escherichia coli hemolysin (HlyA) and Staphylococcus aureus alpha-toxin (S . alpha-toxin) on PMN adhesion to cultured HUVEC . Both toxins increased adherence of human PMN to HUVEC in a dose- and time-dependent manner, peaking after 30 min at 0.01 hemolytic units/ml HlyA or 0.5 microg/ml S . alpha-toxin . Pretreatment of HUVEC with anti-P-selectin mAbs or of PMN with anti-CD11b/CD18 mAb reduced HlyA- and S . alpha-toxin-related cell adhesion significantly . Increased P-selectin expression on toxin-treated endothelial cells was demonstrated by cell surface ELISA . Compared with endotoxin, HlyA and S . alpha-toxin did not induce the expression of E-selectin, ICAM-1, or VCAM-1 . FACS analysis showed increased CD11b/CD18 expression on HlyA-, but not on S . alpha-toxin-stimulated PMN . Platelet-activating factor, an important costimulatory factor for PMN adhesion and activation, was also active in the exotoxin-stimulated adhesion system, as evidenced by studies using the platelet-activating factor receptor antagonist BN50727 . HPLC analysis of endothelial cell extracts confirmed enhanced toxin-mediated PAF synthesis . The capacity of exotoxins to stimulate PMN adhesion to endothelial cells may be relevant in patients with severe local or systemic bacterial infections. Infect Immun, 1996 Nov, 64(11), 4520 - 4 Antibacterial activity of antileukoprotease; Hiemstra PS et al.; Antileukoprotease (ALP), or secretory leukocyte proteinase inhibitor, is an endogenous inhibitor of serine proteinases that is present in various external secretions . ALP, one of the major inhibitors of serine proteinases present in the human lung, is a potent reversible inhibitor of elastase and, to a lesser extent, of cathepsin G . In equine neutrophils, an antimicrobial polypeptide that has some of the characteristics of ALP has been identified (M . A . Couto, S . S . L . Harwig, J . S . Cullor, J . P . Hughes, and R . I . Lehrer, Infect . Immun . 60:5042-5047, 1992) . This report, together with the cationic nature of ALP, led us to investigate the antimicrobial activity of ALP . ALP was shown to display marked in vitro antibacterial activity against Escherichia coli and Staphylococcus aureus . On a molar basis, the activity of ALP was lower than that of two other cationic antimicrobial polypeptides, lysozyme and defensin . ALP comprises two homologous domains: its proteinase-inhibitory activities are known to be located in the second COOH-terminal domain, and the function of its first NH2-terminal domain is largely unknown . Incubation of intact ALP or its isolated first domain with E . coli or S . aureus resulted in killing of these bacteria, whereas its second domain displayed very little antibacterial activity . Together these data suggest a putative antimicrobial role for the first domain of ALP and indicate that its antimicrobial activity may equip ALP to contribute to host defense against infection. Eur J Nucl Med, 1996 Nov, 23(11), 1536 - 9 Retention of technetium-99m in infectious foci in rats after release from technetium-99m labelled human non-specific polyclonal immunoglobulin G: a dual-label study with hydrazinonicotinamido and iminothiolano immunoglobulin; Claessens RA et al.; In an effort to contribute to the understanding of the mechanism of uptake of technetium-99m labelled non-specific polyclonal human immunoglobulin G (hIgG) in inflammatory lesions we compared the tissue distribution of double-labelled 99mTc-hydrazinonicotinamido (HYNIC) hIgG-14C and 99mTc-iminothiolano hIgG-14C in groups of five Wistar rats with a Staphylococcus aureus infection of the left calf muscle between 2 h p.i . and 24 h p.i . The stability of the two double-labelled hIgG preparations was evaluated in vitro and in plasma in vivo by high-performance liquid chromatography (HPLC) analysis . At 24 h after injection of 99mTc-HYNIC-hIgG-14C the abscess uptake of 99mTc (1.5% ID/g+/-0.2% ID/g) was significantly higher (P<0.01) than the 14C uptake (1.0% ID/g+/-0.1% ID/g) . After injection of 99mTc-iminothiolano hIgG-14C no significant difference (P=0.08) was found between the abscess uptake of the two radionuclides at 24 h p.i . (99mTc: 0.8% ID/g+/-0.1% ID/g; 14C: 0.90% ID/g+/-0.09% ID/g) . HPLC analysis of plasma samples revealed release of 99mTc from both double-labelled immunoglobulin preparations . This phenomenon was more pronounced for iminothiolano hIgG than for HYNIC hIgG (43% vs 18%) . In most tissues other than abscesses significant differences were also found between the 99mTc and the corresponding 14C uptake . Our results demonstrate that the chemical form in which 99mTc is bound to hIgG severely influences its release from hIgG and its retention in infections. Eur J Nucl Med, 1996 Nov, 23(11), 1531 - 5 Different behaviour of radioiodinated human recombinant interleukin-1 and its receptor antagonist in an animal model of infection; van der Laken CJ et al.; Recently, we demonstrated that radiolabelled interleukin-1alpha (IL-1) specifically accumulates in focal infection in mice through interaction with its receptor . Unfortunately, systemic side-effects of IL-1 limit its clinical application . We investigated whether this problem could be circumvented by using the interleukin-1 receptor antagonist (IL-1ra), an equally sized protein that binds to the same receptors as IL-1 without induction of biological effects . Biodistribution of 125I-IL-1 and 125I-IL-1ra was determined in Swiss mice with Staphylococcus aureus-induced abscesses in the left calf muscle at 4, 12, 24 and 48 h after injection of either 0.4 MBq 125I-IL-1 or 0.4 MBq 125I-IL-1ra . In vitro, the proteins displayed similar binding characteristics . High-performance liquid chromatographic analysis revealed a tendency for IL-1ra to associate with serum proteins . Both proteins rapidly cleared from most organs . However, the abscess uptake of 125I-IL-1ra was significantly lower than that of 125I-IL-1 at all time points (48 h p.i.: 0.06+/-0 . 01%ID/g vs 0.60+/-0.04%ID/g; P<0.02) . The abscess-to-contralateral muscle ratios did not exceed 15.5+/-2.9 for 125I-IL-1ra, while the ratios for 125I-IL-1 reached 46.9+/-5.7 at 48 h p.i . Despite similar in vitro receptor binding, the abscess uptake of IL-1ra was much lower than that of IL-1 . The interaction of IL-1ra with serum proteins in vivo may reduce its availability for receptor binding in the infection . Although on theoretical grounds IL-1ra is very interesting, these characteristics will prevent its development as a clinically useful radiopharmaceutical to image infection. Gene, 1996 Oct 31, 178(1-2), 97 - 106 NBP35 encodes an essential and evolutionary conserved protein in Saccharomyces cerevisiae with homology to a superfamily of bacterial ATPases; Vitale G et al.; We have cloned a novel and essential gene, NBP35, from Saccharomyces cerevisiae that encodes a putative Nucleotide Binding Protein of 35 kDa . Sequence analysis revealed structural homology of Nbp35p with a family of bacterial ATPases involved in cell division processes and chromosome partitioning . A search in databases identified closely related sequences from yeast and higher eukaryotes, suggesting a conserved function for this family of proteins . By indirect immunofluorescence, a tagged version of Nbp35p carrying two immunoglobulin G-binding domains derived from Staphylococcus aureus Protein A was localised to the nucleus . A single amino-acid substitution in the conserved nucleotide-binding motif of Nbp35p renders the protein non-functional . Furthermore, a conserved cluster of four cysteines in the N-terminal end of the protein is also required for an essential role of Nbp35p. J Immunol Methods, 1996 Oct 30, 198(1), 67 - 77 Chicken anti-protein A prevents Staphylococcus aureus protein A from binding to human and rabbit IgG in immunoassays and eliminates most false positive results; Hoffman WL et al.; This report demonstrates that chicken anti-protein A can prevent both soluble and surface-bound Staphylococcal protein A from binding to either human or rabbit IgG . In an ELISA assay, chicken anti-protein A prevented > 98% of the soluble protein A from binding to the human IgG-Fc coat . In a blotting assay, chicken anti-protein A prevented the membrane-bound protein A from interacting with the human IgG probe . When intact S . aureus (Cowan I strain) was bound to the surface of a microassay plate, chicken anti-protein A blocked > 98% of the cell wall protein A and permitted the probing of the surface components with human IgG . In another immunoassay, rabbit anti-enterotoxin A IgG was used to measure enterotoxin A concentrations in S . aureus culture medium supernatants after soluble protein A was blocked by chicken anti-protein A . Thus, the binding of chicken anti-protein A to protein A almost completely eliminates false positive results and permits the measurement of specific antibodies or antigens in a variety immunoassays where protein A is present. Ann N Y Acad Sci, 1996 Oct 25, 797, 285 - 7 Prophylaxis with the immunomodulator PGG glucan enhances antibiotic efficacy in rats infected with antibiotic-resistant bacteria; Tzianabos AO et al.; The emergence of multiple antibiotic-resistant microorganisms has led to a search for alternatives to traditional therapeutic regimens . PGG glucan is a soluble beta-glucan immunomodulator that selectively enhances the microbicidal activities of neutrophils and macrophages without stimulating proinflammatory cytokine production . In the present studies, we examined the ability of PGG glucan to act in concert with antibiotics to decrease mortality in a rat model of intraabdominal sepsis using antibiotic-resistant bacteria as infectious inocula . Results of these studies demonstrated that prophylaxis with PGG glucan in combination with antibiotics provided enhanced protection against lethal challenge with Esherichia coli or Staphylococcus aureus as compared with the use of antibiotics alone. Presse Med, 1996 Oct 19, 25(31), 1441 - 6 {Nosocomial pneumonia in intensive care units}; Fagon JY et al.; Nosocomial pneumonia is associated with substantial morbidity and mortality . Patients treated with mechanical ventilation have the highest risk for developing this intensive care unit acquired infection . Gram-negative bacilli are the predominant organisms responsible for pneumonia in this setting . However, Staphylococcus aureus has recently emerged as a significant isolate . Nosocomial pneumonia is difficult to diagnose clinically in ventilated patients because fever, lung infiltrate on chest X-ray, leukocytosis are frequent in severely ill patients under mechanical ventilation whatever lung infection is present or not and because lower respiratory tract of such patients is colonized by potentially pathogenic bacteria independently of the presence of true lung infection; thus, different diagnostic strategies are proposed . Our personal bias is that using bronchoscopic techniques to obtain bronchoalveolar lavage and protected-brush specimens permits us to devise a therapeutic strategy that is superior to one based only on clinical evaluation . Measures for prevention of nosocomial infection are essential to decrease the incidence of nosocomial pneumonia and the emergence of multiresistant pathogens. J Immunol, 1996 Oct 15, 157(8), 3298 - 304 Enterotoxin septic shock protection and deficient T helper 2 cytokine production in growth hormone transgenic mice; Gonzalo JA et al.; Neuroendocrine hormones have long been thought to play a role in lymphoid development and function . In particular, growth hormone has been shown to mediate thymic development as well as to promote T cell engraftment in severe combined immunodeficiency mice . Murine T helper cells are classified into two subsets based on their cytokine production pattern . Here, we report that transgenic mice for bovine growth hormone show significant alterations in T cell function and decreased capability for cytokine production, an effect that is more acute in T helper cells as measured by their inability to produce IL-4 upon in vivo injection with Staphylococcus aureus enterotoxin B . Furthermore, upon immunization with conventional Ags, growth hormone transgenic mice produce an altered Ig isotype pattern characterized by a response shift from IgG1 in nontransgenic mice to IgG2 in transgenic mice . The impaired T cell responses correlated with survival from septic shock mediated by bacterial enterotoxins . We conclude that growth hormone may have the potential of regulating immune responses in pathologic processes associated with hyperactivation of T cells or with massive cytokine production. J Am Vet Med Assoc, 1996 Oct 15, 209(8), 1457 - 63 Evaluation of udder health and mastitis in llamas; Rowan LL et al.; OBJECTIVE: To investigate intramammary infections in llamas, identify the pathogens responsible, and determine whether effects of intramammary infection could be detected by use of mastitis indicator tests commonly used for cows . DESIGN: Observational study . ANIMALS: 100 llamas on 10 farms . PROCEDURE: Milk samples were evaluated by bacterial culturing and by determination of somatic cell count (SCC), using direct microscopic and automated counting methods, California Mastitis Test score, pH, and N-acetyl-beta-d-glucosaminidase activity . Correlation coefficients were determined among the various mastitis indicator tests, and test results were determined for milk from infected and uninfected glands . RESULTS: Evidence of intramammary infection was evident in 76 of 369 (21%) milk samples, with 54 of 94 (57%) llamas having at least 1 infected gland . Staphylococcus sp other than Staphylococcus aureus were the predominant pathogens . None of the llamas had clinical signs of mastitis, and significant differences were not detected in SCC, California Mastitis Test score, pH, or N-acetyl-beta-d-glucosaminidase activity between infected and uninfected samples . California Mastitis Test scores were negative or trace for 307 of 313 (98%) samples, and SCC were low . In contrast, pH and N-acetyl-beta-d-glucosaminidase activity of milk from uninfected glands were higher than values reported for milk from uninfected cows, and neither variable was significantly correlated with the number of somatic cells in samples of llama milk . CLINICAL IMPLICATIONS: Although intramammary infections develop in llamas, inflammation (mastitis) appears to be rare . Values for mastitis indicator tests used for cows cannot be directly extrapolated to llamas . Subclinical mastitis is apparently not an important problem in llamas in the United States. J Toxicol Environ Health, 1996 Oct 11, 49(2), 127 - 43 Reaction of human hemoglobin toward the alkylating agent S-(2-chloroethyl)glutathione; Erve JC et al.; In order to investigate if hemoglobin might serve as a biomarker of exposure for 1,2-dichloroethane (DCE) encountered in the workplace, human hemoglobin was alkylated at physiologic pH by the episulfonium ion of S-(2-chloroethyl)glutathione (CEG) . In vitro alkylation resulted in three alkylation products on the alpha chain and at least two alkylation products on the beta chain as determined directly by matrix-assisted laser desorption-ionization mass spectrometry . To ascertain if the site of alkylation was the reactive sulfhydryl present at cysteine-93 on the beta chain of hemoglobin (beta-93 Cys), a spectrophotometric assay using 4,4'-dithiodipyridine was used to measure the free sulfhydryl groups before and after treatment of hemoglobin with various amounts of CEG . Results indicate that the episulfonium ion did not react substantially at beta-93 Cys, as there was no measurable decrease in the sulfhydryl to hemoglobin ratio, even with a large excess of CEG . In contrast, iodoacetamide did react with the sulfhydryl groups and gave a dose-dependent decrease in the sulfhydryl to hemoglobin ratio as measured by this assay . CEG-treated hemoglobin was digested with Staphylococcus aureus endoproteinase Glu-C and the digest was analyzed by fast atom bombardment mass spectrometry . Only one peak in the FAB mass spectrum could correspond to a peptide modified by the episulfonium ion of CEG . These results indicate that although the episulfonium ion of CEG does alkylate human hemoglobin, beta-93 Cys is not the major alkylation target. Microb Drug Resist, 1996 Fall, 2(3), 343 - 51 Testing the efficacy of a molecular surveillance network: methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecium (VREF) genotypes in six hospitals in the metropolitan New York City area . The BARG Initiative Pilot Study Group . Bacterial Antibiotic Resistance Group; de Lencastre H et al.; Molecular fingerprinting techniques are rapidly becoming indispensable tools for hospital epidemiology . On the other hand, the relative complexity and unfamiliarity of these techniques to most hospital diagnostic laboratories limit their usefulness . In an attempt to provide a solution for this dilemma, we tested the feasibility and efficacy of a cooperative venture in which molecular typing of isolates recovered from patients in six hospitals was performed at two microbiology research laboratories with expertise in these techniques . In a small preliminary study, 30 methicillin-resistant Staphylococcus aureus (MRSA) and 30 vancomycin-resistant Enterococcus faecium (VREF) isolates were collected over a 3-week period from six hospitals in the metropolitan New York area and transported to the Laboratory of Microbiology at The Rockefeller University during the summer months of 1994 . Nineteen of the 27 confirmed MRSA isolates were closely related strains carrying the same mecA and the same Tn554 polymorphs in a pulsed-field gel electrophoresis (PFGE) background represented by closely related subtypes of a single pattern, indicating the wide distribution of this MRSA clone among the participating hospitals . Typing of the same 27 MRSA isolates was also performed at the Tuberculosis Center of the Public Health Research Institute and identical results were obtained . The 29 confirmed VREF isolates were highly heterogeneous and belonged to as many as 23 distinct clonal types as defined by PFGE patterns and probing with vanA . Characterization of the 60 isolates by these methods was completed in one month of full-time effort by a single experienced laboratory assistant guided by a doctoral-level expert in molecular fingerprinting techniques . The collection of samples for both MRSA and VREF was not intended to address epidemiological questions but to determine the feasibility of a multicenter study . On the basis of our preliminary findings we are encouraged that a larger cooperative effort is possible and with the correct sampling method we believe that epidemiological and surveillance studies could be accomplished that would provide a tracking system to assist hospitals, clinics, and chronic care facilities in controlling the spread of multidrug-resistant pathogens. Microb Drug Resist, 1996 Fall, 2(3), 331 - 41 Characterization of methicillin-resistant Staphylococcus aureus isolates from Portuguese hospitals by multiple genotyping methods; Aires de Sousa M et al.; One hundred and eighty-three methicillin-resistant Staphylococcus aureus (MRSA) isolates from eight different Portuguese hospitals were genetically typed by random amplification of polymorphic DNA (RAPD) employing different oligonucleotide primers . Fourteen different RAPD genotypes were identified . A subset of the same strains was also characterized by pulsed-field gel electrophoresis (PFGE) and/or hybridization using mecA and Tn554 probes . In the majority of cases, the different genotyping methods have identified the same MRSA clones . However, PFGE combined with the DNA probes was clearly the method providing higher resolution . Most strains that have already been identified by PFGE and DNA probes as members of the widely spread Iberian clone of MRSA generated a common RAPD genotype . The most prevalent Iberian clone was not detected in a collection of MRSA from Poland that was also examined by RAPD . On the other hand, MRSA strains second most frequent in prevalence in the Portuguese and Polish collection appear to be identical by RAPD, indicating extensive geographic spread of this particular clone . No correlation was apparent between epidemic behavior and the number of protein A gene repeats in this particular collection of MRSA strains. Microb Drug Resist, 1996 Fall, 2(3), 319 - 29 Tracing the origin of an outbreak of methicillin-resistant Staphylococcus aureus infections in a Portuguese hospital by molecular fingerprinting methods; Sanches IS et al.; Seventy-six methicillin-resistant Staphylococcus aureus (MRSA) isolates were collected from July 1992 to May 1995 at a 400-bed district hospital in the northeast of Portugal . During the second half of the surveillance period, in July of 1994, an outbreak was detected in the orthopedic ward . Thirty-three (out of the 76) MRSA strains were recovered only in this ward during the outbreak period . All strains were characterized by a variety of genomic fingerprints . Hybridization of ClaI and SmaI restriction digests with the mecA- and Tn554-specific DNA probes was used to identify polymorphism and determine chromosomal location of these determinants, and pulsed-field gel electrophoretic analysis of SmaI digests was used to determine chromosomal backgrounds . All strains recovered during the outbreak in the orthopedic ward were found to belong to a single clone that carried the mecA polymorph I, Tn554 type E in a macrorestriction background called H (clone I::E::H1), which was identified in 18 patients, and 5 health care personnel and from a fomite sample, and was traced to a single transfer patient admitted to the hospital at the beginning of the outbreak . The new clone I::E::H1 differed only in the macrorestriction profile from the MRSA clone previously dominant in this hospital, known as Iberian epidemic clone I::E::A, which has already been identified in several Spanish and Portuguese hospitals. Pneumologie, 1996 Oct, 50(10), 706 - 11 {MRSA (methicillin-resistant Staphylococcus aureus) infections in patients with pulmonary diseases}; Erbes R et al.; Methicillin-resistant Staphylococcus aureus (MRSA) has become a major nosocomial pathogen . We investigated MRSA-infections in patients with pulmonary diseases referring to epidemiological aspects . Between 9/92 and 2/92 we found MRSA-infections in our hospital in 24 patients (11 female, 13 male, average age 54.6 years) . Clinical presentation, main and accompanying disorders and previous antibiotic therapy regimens were registered . Strains were typed using DNA-RFLP and lysotyping . MRSA detection were done in specimen from sputum (12/24) and from the bronchial secret (9/24) . In 18/24 cases the MRSA-colonisation was associated with infection . In 15/24 cases the first acquisition of MRSA happened in our hospital, 6/24 times the germ was carried off other institutions and in 3/24 cases it was possibly community acquired . Most frequently patients suffered from bronchial cancer (6/24), from chronical bronchitis (5/24), from pneumonia (4/24) or Cystic fibrosis (4/24) . Usually the patients showed other severe comorbidity . 13/24 patients had an antibiotic course before detecting MRSA . Typing revealed a strain already known in different hospitals of Berlin, another known strain of northern Germany and two so far unknown strains . Of interest was a different behaviour of resistance and the lost of resistance of strains in the course . MRSA-infection in pulmonary medicine emerged as a problem mostly in patients with multimorbidity and severe underlying diseases . Change of resistance in strains and new strains were observed. Monaldi Arch Chest Dis, 1996 Oct, 51(5), 387 - 90 Mixed infection by Staphylococcus and Candida, and Wegener's granulomatosis; Valenti S et al.; We describe the case of a patient who initially presented with pneumonia from Staphylococcus aureus and Candida parapsilosis, which was resolved with antibiotic treatment, but reappeared 6 months later as full-blown Wegener's granulomatosis . The possible pathogenetic correlations between infective agents, in particular Staphylococcus aureus and Candida, and Wegener's granulomatosis are discussed. Biosci Biotechnol Biochem, 1996 Oct, 60(10), 1571 - 4 Detection of protein A produced by Staphylococcus aureus with a fiber-optic-based biosensor; Chang YH et al.; Staphylococcus aureus is a pathogen important in causing human infections and intoxication . A sensitive fiber-optic that produces evanescent waves was developed for the detection of protein A, a product secreted only by S . aureus . In the immunosensor, a 40-mV argon-ion laser that generated laser light at 488 nm was used together with plastic optical fiber and antibodies to protein A were physically adsorbed onto the fiber . The principle of the detection involved a sandwich immunoassay with fluorescein isothiocyanate conjugated with anti-(protein A) immunoglobulin G to produce signals of the antigen-antibody reaction . The detection limit was 1 ng of protein A per milliliter . The fiber-optic immunosensor could be used for rapid and specific detection of S . aureus in clinical specimens and foods. Respir Med, 1996 Oct, 90(9), 531 - 7 Predictors of Staphylococcus aureus pneumonia associated with human immunodeficiency virus infection; Tumbarello M et al.; This study is based on a retrospective logistic regression analysis of all human immunodeficiency virus (HIV)-infected patients with Staphylococcus aureus pneumonia (SAP) admitted to the Department of Infectious Diseases, Catholic University, Rome, Italy between January 1986 and December 1994 . Nineteen patients with 24 episodes of SAP were enrolled in the study . A control group of 38 HIV-infected patients without pneumonia was included . The attack rate of SAP was 8.31/1000 HIV-related hospital admissions and the frequency, out of the total number of bacterial pneumonia observed in the study period, was 16% (24 of 154 patients) . The large majority of SAP was community acquired . On the univariate analysis, intravenous drug abuse (IVDA) (P = 0.02), history of previous Pneumocystis carinii pneumonia (PCP) (P = 0.03) and cirrhosis (P = 0.03) were significant risk factors for SAP . In addition, IVDA and previous PCP were independent risk factors on multivariate analysis . All patients presented with fever associated with cough (74%), chest pain (26%) or shortness of breath (37%) . Chest X-ray documented lobar pneumonia (78%), predominantly in the lower lobes, consolidation with cavitation (11%), and interstitial-nodular infiltrates (11%) . Pleural effusion was present in 31% of patients . The response to therapy was favourable in 79% of patients . Recurrence occurred in 26% and death occurred in 21% of patients . Death was significantly associated with the low level (< 50 mm-3) of circulating T CD4+ cells (P = 0.03) and the recurrence of pneumonia (P = 0.03) . In conclusion, the present study indicates that S . aureus is an important aetiologic agent of bacterial pneumonia in HIV-infected patients, especially if they are drug abusers with previous PCP. Vet Immunol Immunopathol, 1996 Oct, 53(3-4), 249 - 56 In vitro responses of cervine macrophages to bacterial stimulants; Cross ML et al.; The function of cervine (deer) mononuclear phagocytes is poorly defined . In the present study, the potential of cervine macrophages to generate phagocytic and immunoregulatory responses following stimulation with bacterial products was investigated . Blood-derived macrophages of red deer were cultured in vitro with particulate stimulants (Mycobacterium bovis BCG and Staphylococcus aureus SAC) or soluble stimulants (M . bovis PPD and Escherichia coli LPS), prior to assessment of phagocytic responses, prostaglandin secretion and cytokine production . Particulate stimulants induced vigorous phagocytic responses (superoxide anion generation, lysosomal enzyme release), secretion of prostaglandin E2 and transcription of mRNA specific for the cytokines IL-1 beta, IL-10 and TNF alpha, while soluble products invoked weaker responses . These results are discussed in relation to the role of cervine mononuclear phagocytes in regulating and participating in inflammatory and immune processes relevant to bacterial challenge. J Antimicrob Chemother, 1996 Oct, 38(4), 589 - 97 The concentration-independent effect of monoexponential and biexponential decay in vancomycin concentrations on the killing of Staphylococcus aureus under aerobic and anaerobic conditions; Larsson AJ et al.; An in-vitro pharmacodynamic system was used to generate time-kill curves to demonstrate the concentration-independent pharmacodynamics of vancomycin against Staphylococcus aureus ATCC 29213 . Initial vancomycin concentrations of 5, 10, 20 and 40 mg/L were studied monoexponentially while simulating a 6 h half-life . One parallel experiment was performed in duplicate using an initial peak concentration of 40 mg/L where both a distribution alpha-phase half-life of 0.66 h for 1 h and an elimination beta-phase half-life of 6 h for 11 h were simulated to determine if the transient distribution phase concentrations of vancomycin have any impact on bacterial killing beyond that provided by the elimination phase concentrations . Additionally, two monoexponential experiments with peak concentrations of 40 and 20 mg/L and a half-life of 6 h were repeated in an anaerobic chamber to determine if killing of S . aureus was affected . The time to achieve a 3 log10 kill was calculated from the linear portion of the regression line and averaged (mean +/- S.D.) 9.0 +/- 1.4 h for all aerobic monoexponential experiments and was 8.4 and 8.6 h for the aerobic biexponential experiments (P > 0.05) . For the anaerobic studies, the times to reach 3 log10 kill were significantly greater averaging 18.9 +/- 1.7 h . The slopes of the bacterial kill curves were virtually identical for both monoexponential and biexponential aerobic experiments averaging -0.34 +/- 0.04, yet significantly different from the anaerobic bacterial kill curve slopes of -0.16 +/- 0.015 (P < 0.05) . Time-kill curve analyses suggest that varying the concentration of vancomycin does not affect the rate or extent of bacterial killing aerobically or anaerobically against S . aureus and more efficient killing was achieved under aerobic conditions . The simulated distribution phase concentrations did not contribute to more effective killing of this strain of S . aureus. Infect Immun, 1996 Oct, 64(10), 4288 - 98 Altered regulation of inducible nitric oxide synthase expression in macrophages from senescent mice; Chen LC et al.; We investigated the capacity of mouse macrophages obtained from senescent animals to respond in vitro to microbial stimuli . Significant hypersecretion of nitric oxide (NO) was observed in thioglycolate-elicited macrophages from senescent mice compared with those obtained from young mice in response to lipopolysaccharide (LPS) . In contrast, both cell populations manifested equivalent responses to LPS with respect to tumor necrosis factor alpha secretion . Further, macrophages from senescent animals also showed potentiated responses to both zymosan and heat-killed Staphylococcus aureus, as assessed by NO production . Both cell populations were equivalently inhibited by a competitive inhibitor of NO synthase NG-monomethyl-L-arginine . Since endogenous beta interferon (IFN-beta) is recognized as an essential cofactor for LPS-induced NO production by macrophages, we investigated the role of IFN-beta in enhancing the capacity of both macrophage populations for LPS-induced NO production . Macrophages from young mice were minimally activated by LPS alone to express inducible NO synthase (iNOS), and the response was significantly potentiated by the addition of IFN-beta . These findings were confirmed by immunocytochemical staining of iNOS in which the frequency of iNOS-positive cells in response to LPS was enhanced in the presence of IFN-beta . Reverse transcription-PCR analyses revealed that macrophages from senescent animals produced larger amounts of iNOS mRNA in response to LPS . Further, exogenous IFN-beta potentiated iNOS mRNA expression in macrophages from young mice . In contrast, the frequency of LPS-activated macrophages for iNOS expression was markedly increased during senescence and addition of IFN-beta did not significantly change this frequency . These results correlated with reverse transcription PCR data showing high levels of iNOS mRNA in LPS-stimulated macrophages from senescent mice . LPS-induced NO production in macrophages from both young and senescent mice was inhibited by neutralizing antibody to either IFN-beta or IFN-gamma . Mixed cultures of macrophages from young and senescent mice stimulated with LPS manifested significantly enhanced NO production relative to that which would be predicted from an additive response of the two macrophage populations stimulated separately . The differential responsiveness of NO production observed with thioglycolate-elicited macrophages from young and senescent mice was also observed in resident macrophages but, interestingly, not in bone marrow culture-derived macrophages . These results suggest that environmental factors may be responsible for the potentiated NO responses of macrophages from senescent mice . Collectively, these data suggest that macrophages from senescent animals manifest an altered mechanism for regulation of macrophage function in NO production and iNOS expression by constitutive and/or induced expression of autoregulatory cytokines. Infect Immun, 1996 Oct, 64(10), 4231 - 5 Gastrointestinal colonization by methicillin-resistant Staphylococcus aureus in immunosuppressed mice; Kato-Matsunaga N et al.; ICR mice were inoculated intranasally with methicillin-resistant Staphylococcus aureus (MRSA) N133, and the inoculated MRSA was quantitatively recovered from the ceca and feces . The viable counts of the MRSA recovered from ceca correlated well with those from feces . Some mice eliminated MRSA from the cecum by 14 days after inoculation . Intraperitoneal administration of cyclophosphamide at a dose of 200 mg/kg 3 days before inoculation inhibited the elimination of the MRSA from both ceca and feces . All mice treated with cyclophosphamide excreted more than 10(4) CFU of the MRSA per g of feces for at least 70 days, indicating persistent colonization of the MRSA in the gastrointestinal tract . Some beta-lactam antibiotics decreased the colonization level, but others did not . The colonization level was suggested to depend on the antibacterial activity of the antibiotic against the MRSA and the degree of disturbance of intestinal flora by the antibiotic. Zentralbl Veterinarmed B, 1996 Oct, 43(8), 475 - 82 Microbiological diagnoses of chronic otitis externa in the dog; Blanco JL et al.; The microbiological characteristics of otic exudates from 26 dogs with chronic otitis externa was studied with special reference to the implication of yeasts in the aetiology of the disease . A high frequency of yeasts and Staphylococcus aureus were isolated, alone or in association . In reference to the yeasts, there was a clear predominance of the genus Candida (48% of the total yeasts) . Malassezia (Pytirosporum) represented only 3% of the isolates . It can be concluded that yeasts have an important role in the pathogenicity of this disease . For the microbiological diagnosis of otitis externa, we recommend the simultaneous use of Columbia/5% Sheep Blood Agar and Sabouraud-Dextrose without antibiotic addition, the use of 37 degrees C as the incubation temperature and direct microscopic observation of the sample before culture. Int Immunol, 1996 Oct, 8(10), 1603 - 7 Linomide, an immunomodulator that inhibits Th1 cytokine gene expression; Arad G et al.; Linomide (LS-2616, quinoline-3-carboxamide) has strong immunomodulating effects in animal models, inhibiting toxic shock, progressive autoimmune disease and cancer . In humans, linomide strongly reduced the appearance of new lesions in multiple sclerosis yet enhanced immune responses after bone marrow transplantation . In contrast to these clear effects in vivo, attempts to show an effect of linomide in vitro have not been successful and its mode of action remains to be elucidated . Here we show that at concentrations effective in vivo, linomide is active on human peripheral blood mononuclear cells (PBMC), severely inhibiting the induction by Staphylococcus aureus enterotoxin B of mRNA of three cytokine genes expressed in Th1 cells, those for IFN-gamma, IL-2, and tumor necrosis factor-beta . Yet, cell viability was not affected by linomide . The extent of inhibition is dose-dependent on linomide . Linomide also blocked induction of IL-2 and IFN-gamma mRNA by phytohemagglutinin . The inhibitory effect is expressed immediately but can be enhanced significantly by a prolonged exposure of PBMC to linomide, reaching 10-fold . These results support the concept that linomide antagonizes the activation of Th1 cells during a cellular immune response. Br J Dermatol, 1996 Oct, 135(4), 528 - 32 Antimicrobial effects of phototherapy and photochemotherapy in vivo and in vitro; Yoshimura M et al.; We investigated the antimicrobial effects of phototherapy and photochemotherapy in vivo and in vitro . First, Staphylococcus aureus samples were obtained using stamp agar medium from inflammatory lesions of 29 adult patients with atopic dermatitis before and after a single photochemotherapy . Therapy was oral PUVA (30 mg 8-methoxypsoralen, 8MOP plus 5J/cm2 UVA), topical PUVA (0.3% 8MOP plus 200 mJ/cm2 UVA) or UVB (80 mJ/cm2) irradiation . The number of S . aureus on the lesions was significantly reduced, even after a single treatment with all therapies . Reductions (mean +/- SD) were 69.3 +/- 26.9%, 76.3 +/- 31.3% and 83.8 +/- 18.5%, respectively . Secondly, we investigated the effect of PUVA (0.001% 8MOP plus 10, 20, 30, 40, or 50 mJ/cm2 UVA) and UVB (10, 30, 50, or 100 mJ/cm2) irradiation on the proliferation of S . aureus in vitro . PUVA and UVB treatment markedly inhibited the proliferation in a dose-dependent manner . These results seem to indicate the possibility that the antimicrobial effect of UV radiation contributes to successful photochemotherapy in patients with atopic dermatitis. Adv Ren Replace Ther, 1996 Oct, 3(4), 302 - 8 Staphylococcus aureus vaccination for dialysis patients--an update; Fattom AI et al.; Staphylococcus aureus infections are a major cause in both hemodialysis and peritoneal dialysis patients . The availability of a safe and effective protective vaccine would be of great benefit to these patients, but attempts at using vaccines consisting of inactivated whole cells have been unsuccessful . This article discusses an alternate approach to S . aureus vaccine design using a capsular polysaccharide conjugate and preliminary results in hemodialysis and peritoneal patients. J Hosp Infect, 1996 Oct, 34(2), 151 - 60 Epidemiology of methicillin-resistant Staphylococcus aureus in a Warsaw hospital; Mlynarczyk G et al.; Methicillin-resistant Staphylococcus aureus (MRSA) isolates were collected during two eight-month periods in 1991 and 1994, respectively . In order to study the epidemiology, all 74 strains were characterized by phage-typing, antibiotic resistance patterns and DNA-restriction map after cleavage with SmaI enzyme, and pulsed-filed gel electrophoresis (PFGE) . These investigations confirmed that MRSA in the hospital, 1991 and 1994, was not due to the spread of one or two clones, but by the simultaneous occurrence of a few well characterized strains and sporadic, occurring strains of different phage-types . Some of these might have developed from the more commonly occurring strains . Isolates from 1994 were more resistant to antibiotics in vitro, than the 1991 isolates . The typing results also indicated that whilst most of the MRSA strains in 1994 were different compared with those of 1991, some of the strains might have been present in both years . The PFGE-typing was more discriminatory and gave a higher typability than the phage-typing, especially among the multiply resistant isolates of MRSA from 1994 . Among the less resistant strains the phage-typability was high and with only few exceptions, there was a good correlation between PFGE-type and phage-type. J Hosp Infect, 1996 Oct, 34(2), 145 - 9 Survival of methicillin-resistant Staphylococcus aureus (MRSA) on naturally contaminated dry mops; Oie S et al.; The floors of single rooms being used by inpatients colonized by methicillin-resistant Staphylococcus aureus (MRSA) were cleaned using disposable dust-attracting dry mops . Each mop was divided into 12 sections and MRSA quantified serially . This experiment was repeated a total of 21 times for four patients . The MRSA survival rate on the dry mops compared with a control was 59.0-125% after seven days, 26.3-41.6% after 14 days, 0.1-16.2% after 28 days, 0-0.1% after 56 days and 0% after 84 days . MRSA disseminated by patients over the environment can survive for several weeks on dry mops. Pediatr Pulmonol, 1996 Oct, 22(4), 236 - 47 Acute bronchiolitis in tropical Africa: a hospital-based perspective in Ibadan, Nigeria; Johnson AW et al.; In a 30-month prospective study of severe acute lower respiratory infections in hospitalized pre-school Nigerian children, acute bronchiolitis was diagnosed in 67 cases; 19 (28.4%) and 2 (3.0%) of these had concomitant pneumonia or croup, respectively . The peak prevalence was in the wet (rainy) season (May-October) . The male/female (M:F) ratio in infants < or = 6 months was 2.9:1, differing significantly from the 1.1:1 in older subjects (P = 0.04) . None of the subjects had severe malnutrition . Neither a high fever (> or = 39 degrees C), nor tachypnea on admission was significantly correlated with co-existing pneumonia . Of the 29 subjects in whom it was possible to explore viral immunofluorescence studies and/or serodiagnosis, we identified 26 viral identifications in 18 (62.1%) cases; 6 (20.7%) had > or = 2 viruses . Respiratory syncytial virus was identified in 11 (38.0%) of the 29 cases, and parainfluenza virus (PIV) types 1, 2, and 3 in 10 (34.5%) . PIV type 3 accounted for 7 cases, including 3 with bacteremia . Bacterial isolates were made in 9 (21.4%) of 42 blood cultures and in the only lung aspirate; Staphylococcus epidermidis and Staphylococcus aureus accounted for 4 and 3 cases, respectively . Although bacteremia was 2.9 times more common in cases with co-existing pneumonia or croup, the respective frequency of virus-positive cases and that of bacteremia was not significantly different between cases with bronchiolitis alone and those with associated pneumonia or croup . No deaths were recorded, but subjects aged > 6 months had a significantly shorter hospital stay than those < 6 months old (P = 0.02) . Despite the limited sample size, our findings reflect the etiological importance of the paramyxoviruses and the seasonal pattern of bronchiolitis in tropical Africa. Infect Control Hosp Epidemiol, 1996 Oct, 17(10), 654 - 9 Quantitative antibiogram as a typing method for the prospective epidemiological surveillance and control of MRSA: comparison with molecular typing; Blanc DS et al.; OBJECTIVE: Evaluation of the quantitative antibiogram as an epidemiological tool for the prospective typing of methicillin-resistant Staphylococcus aureus (MRSA), and comparison with ribotyping . METHODS: The method is based on the multivariate analysis of inhibition zone diameters of antibiotics in disk diffusion tests . Five antibiotics were used (erythromycin, clindamycin, cotrimoxazole, gentamicin, and ciprofloxacin) . Ribotyping was performed using seven restriction enzymes (EcoRV, HindIII, KpnI, PstI, EcoRI, SfuI, and BamHI) . SETTING: 1,000-bed tertiary university medical center . RESULTS: During a 1-year period, 31 patients were found to be infected or colonized with MRSA . Cluster analysis of antibiogram data showed nine distinct antibiotypes . Four antibiotypes were isolated from multiple patients (2, 4, 7, and 13, respectively) . Five additional antibiotypes were isolated from the remaining five patients . When analyzed with respect to the epidemiological data, the method was found to be equivalent to ribotyping . Among 206 staff members who were screened, six were carriers of MRSA . Both typing methods identified concordant of MRSA types in staff members and in the patients under their care . CONCLUSIONS: The quantitative antibiogram was found to be equivalent to ribotyping as an epidemiological tool for typing of MRSA in our setting . Thus, this simple, rapid, and readily available method appears to be suitable for the prospective surveillance and control of MRSA for hospitals that do not have molecular typing facilities and in which MRSA isolates are not uniformly resistant or susceptible to the antibiotics tested. Infect Control Hosp Epidemiol, 1996 Oct, 17(10), 649 - 53 Colonization by Staphylococcus aureus resistant to methicillin and ciprofloxacin during 20 months' surveillance in a private skilled nursing facility; Lee YL et al.; OBJECTIVE: To evaluate endemic colonization with Staphylococcus aureus resistant to methicillin, ciprofloxacin, or both among patients of a private skilled nursing facility, with regard to colonization rate and site, and relation to infection and prior antibiotic use . DESIGN: Prospective quarterly culture surveillance of nares and rectal specimens over 20 months' observation . RESULTS: The mean prevalence was 3.8% in new admissions and 5.4% for in-house patients; cumulatively, 7.5% of the patients were colonized during the study period . The colonization rate remained stable during the study period . Screening of rectal, as well as nares, specimens detected substantially more colonized patients than would have been detected by nasal cultures alone . Five to seven percent of the colonized patients developed later infection with methicillin-ciprofloxacin-resistant S aureus . Colonized patients did not differ significantly from the noncolonized group in prior use of quinolones, but the colonized group was exposed significantly more frequently to other antibiotics than the noncolonized group . Eighty-three percent of methicillin-resistant S aureus (MRSA) isolated from infections and 89% from colonization were also ciprofloxacin resistant . CONCLUSION: Although all infecting and most colonizing isolates of MRSA were resistant to quinolones, the overall rate of colonization remained low and stable despite the continued use of quinolones . The findings suggest that good infection control practice has prevented broader spread of such strains in this facility. Can J Microbiol, 1996 Oct, 42(10), 1024 - 31 Staphylococcus aureus strains differ in their in vitro responsiveness to human urokinase: evidence that methicillin-resistant strains are predominately nonresponsive to the growth-enhancing effects of urokinase; Hart DA et al.; Clinical isolates of Staphylococcus aureus were found to exhibit strain-specific heterogeneity to the growth-enhancing effects of human urokinase (UK), a proteinase with plasminogen activator activity . Nine out of fourteen (64%) methicillin-sensitive strains of S . aureus were responsive to UK in "in vitro" cultures . In contrast, 3/29 (10%) methicillin-resistant strains were responsive to the proteinase . When only strains isolated from western Canada were considered, 6/11 methicillin-sensitive strains and 1/26 methicillin-resistant strains were responsive to UK . The single western Canadian methicillin-resistant strain (strain 456) responsive to UK was one of two isolated from the same patient, indicating that the two strains were phenotypically different . Strain 456, resistant to 32 micrograms methicillin/mL, was responsive to as little as 50 U UK/mL and enhancement of growth was evident by 9 h of incubation at 37 degrees C . This growth enhancement was specific to UK and not duplicated by equivalent concentrations of other proteins (bovine serum albumin, trypsin, plasminogen) . The results presented indicate differences in the frequency of the UK-responsive phenotype between methicillin-sensitive and -resistant S . aureus . These findings indicate that the UK phenotype of S . aureus may have utility in both phenotyping clinical isolates, as well as providing insights into the regulation of growth in this clinically important organism. Biotechnol Appl Biochem, 1996 Oct, 24 ( Pt 2), 155 - 9 Expression and purification of recombinant toxicshock-syndrome toxin I; Wahlsten JL et al.; Toxic-shock-syndrome toxin I (TSSTI), an exotoxin produced by certain strains of Staphylococcus aureus, has been closely associated with the pathogenesis of toxic shock syndrome . Outside the context of its staphylococcal host, TSSTI may offer therapeutic uses . We report here a strategy for high-level expression and simplified purification of TSSTI . We have subcloned the coding region for TSSTI into a vector containing an inducible T7 promoter sequence and expressed the protein in an Escherichia coli host strain . The recombinant TSSTI protein contained ten sequential histidine residues (Histag) at its N-terminus, which enabled its efficient purification using nickel-agarose-affinity resin . Histag-TSSTI (H-TSSTI) was further purified to homogeneity using a size-exclusion column . By this system, 80 mg of highly purified H-TSSTI can be consistently obtained per litre of culture in under 3 days . H-TSSTI retained biological activity and was unaffected by the presence of the Histag, as measured in lymphocyte proliferation assays. Arch Phys Med Rehabil, 1996 Oct, 77(10), 1014 - 8 Clinical use of percutaneous intramuscular electrodes for functional electrical stimulation; Shimada Y et al.; OBJECTIVE: To evaluate the clinical use of the percutaneous intramuscular electrode in functional electrical stimulation (FES) . DESIGN: Randomized and controlled study . SETTING: A referral center and institutional practice providing outpatient care . PATIENTS: Seventeen patients (12 men, 5 women) who had implanted percutaneous intramuscular electrodes for more than 1 year were examined . The average follow-up time after implantation of electrodes was 2.2 years (range, 1yr to 4yr 10mo) . Overall, there were 327 electrodes (83 upper extremities and 244 lower extremities) . INTERVENTION: The indwelling electrode was composed of helically coiled Teflon-coated rope stranded from 19 hard drawn wires of SUS 316L stainless steel (SES 114) . MAIN OUTCOME MEASURES: The rates of breakage, movement, and infection, and the number of electrodes that needed reimplantation were evaluated . RESULTS: Only one electrode broke (0.3%) in the iliopsoas muscle at 12 weeks after implantation . Eight electrodes (2.4%) were removed because of loss of sufficient contraction force caused by movement of the electrodes . Movements occurred at 9 weeks in 6 electrodes and at 5 months in two . The failure rate of electrodes in the lower extremities was 3.7% . No failures occurred in the upper extremities . Ten electrodes (3.1%) required reimplantation . Although ten superficial infections (3.1%) were seen around the site of electrode insertion, no removals of electrode were needed . All electrodes in one patient were removed, however, because of generalized methicillin-resistant Staphylococcus aureus infection complicated with renal disease . Electrodes were reimplanted after improvement of the infection . CONCLUSIONS: The ultrafine percutaneous intramuscular electrode was considered practical for long-term FES use. J Med Microbiol, 1996 Oct, 45(4), 294 - 301 Persistence of Staphylococcus aureus strains among cystic fibrosis patients over extended periods of time; Branger C et al.; Pulsed-field gel electrophoresis (PFGE) of SmaI macrorestriction fragments of chromosomal DNA was used to confirm the persistence of methicillin-sensitive Staphylococcus aureus isolates in the sputum of 25 cystic fibrosis patients in five French hospitals . Three-to-eight consecutive isolates, with the same esterase electrophoretic type isolated from each patient over a period of 12-28 months, were analysed . Consecutive isolates with indistinguishable PFGE profiles were found in 12 patients (48%) and consecutive isolates with similar PFGE profiles showing minor differences of one-to-four fragments (similarity coefficient >/=84%) were found in 11 patients . Consecutive isolates with different PFGE profiles were obtained from only two patients, but the profiles found in each patient were more closely related to each other than to other profiles . The results were in agreement with esterase electrophoretic typing for 23 patients, and we considered that those patients were infected with a single persistent strain . For any given patient, variations in antibiotypes and phage types of consecutive isolates were not associated with major genotypic variations . PFGE is useful in confirming the persistence of S . aureus strains in cystic fibrosis patients over long periods. Scand J Immunol, 1996 Oct, 44(4), 361 - 8 Increasing cytotoxic activity and production of reactive oxygen and nitrogen intermediates by peritoneal macrophages during the development of multiple organ dysfunction syndrome in mice; Jansen MJ et al.; A major problem in the intensive care unit nowadays is the development of multiple organ dysfunction syndrome (MODS), a cumulative sequence of progressive deterioration of organ functions . While the pathogenic pathways of MODS remain to be elucidated, it is assumed that cells of the host defence system, especially the macrophages, are altered in their function . During the development of MODS it is assumed that macrophages are overactivated and that an exaggerated inflammatory response may contribute to its pathogenesis . In order to gain insight into the alterations of the functional status of the macrophage during the development of MODS, a series of macrophage functions was measured in the subsequent phases of zymosan induced generalized inflammation in mice . Male C57BL/6 mice received a single dose of zymosan intraperitoneally and groups of animals were killed after 2, 5, 8, and 12 days . Peritoneal macrophages were collected for in vitro assessment of the ADCC, the production of superoxide (O2-) and nitric oxide (NO), and complement mediated phagocytosis and intracellular killing of Staphylococcus aureus . A single intraperitoneal injection with zymosan resulted in a three-phase illness . During the third phase the animals developed MODS-like symptoms . Peritoneal cells from control animals produced very low to non-detectable amounts of O2- and NO, and the cytotoxic activity was also low . During the development of MODS, from day 7 onwards, the ability to produce O2- and NO2- became strongly elevated, as did the cytotoxic activity . These findings are in parallel with the development of MODS whereas the phagocytic and killing capacity remained essentially unaltered . The changes found could be detrimental for the organism, thus possibly contributing to the onset and development of MODS. J Infect Dis, 1996 Oct, 174(4), 800 - 5 Defective polymorphonuclear leukocyte functions in children receiving chemotherapy for cancer are partially restored by recombinant human granulocyte colony-stimulating factor in vitro; Lejeune M et al.; Granulocyte colony-stimulating factor (G-CSF) has important direct and priming effects on different functions of normal mature polymorphonuclear leukocytes (PMNL) . Previous study has shown an alteration in respiratory burst and bactericidal activities of PMNL harvested from children with cancer treated with chemotherapy . The present study evaluates the possibility that recombinant human (rh) G-CSF could correct these defective functions in vitro . Free radical formation in defective PMNL was enhanced by rhG-CSF to a level similar to that found in normal PMNL primed by rhG-CSF . The defective bactericidal activity against Escherichia coli and Staphylococcus aureus was also corrected . This bactericidal activity was not different from that observed in normal PMNL primed by rhG-CSF . In conclusion, correction of the altered free radical-formation pathway by rhG-CSF in these cells contributed to the restoration of normal bactericidal activity against both gram-positive and gram-negative microorganisms. FEMS Microbiol Lett, 1996 Oct 1, 143(2-3), 203 - 10 Localization of prophages of serological group B and F on restriction fragments defined in the restriction map of Staphylococcus aureus NCTC 8325; Borecka P et al.; Several Staphylococcus aureus strains were lysogenized by the phages of serological group B (phages phi 53, phi 85) as well as by some of serological group F (phages phi 77, phi 84) and macrorestriction fragment patterns of genomic DNA were estimated in the lysogenized, non-lysogenic and delysogenized (cured of prophages) strains . It was shown that the integration of phage DNA into chromosome of S . aureus leads to specific changes in restriction fragment pattern in all the lysogenized strains . These changes correlate well with the SmaI restriction map of S . aureus NCTC 8325 since they concern the restriction fragments defined in this map . Phages phi 53 and phi 85 integrate into SmaI fragment B . On the other hand, phages phi 77 and phi 84 integrate into SmaI fragment E of the S . aureus restriction map . The prophages of strain NCTC 8511 have their integration sites, as follows: the phage designated by us phi M integrates in fragment A, whereas the integration site for phage phi J lies in fragment E . Phage phi M was estimated to be genetically related to phages of serological group A and phage phi J to those of serological group F . Evidence was given that lysogenization of S . aureus strains by at least four prophages does not cast any doubt upon the estimation of their genetic relatedness based on their similarity in restriction pattern. FEMS Microbiol Lett, 1996 Oct 1, 143(2-3), 195 - 201 Detection of the response regulator AgrA in the cytosolic fraction of Staphylococcus aureus by monoclonal antibodies; Morfeldt E et al.; The expression of many virulence genes in Staphylococcus aureus is controlled by a regulatory RNA molecule, RNAIII, which is encoded by the agr locus . Transcription RNAIII requires the activity of the agrA, B, C and D genes, which code for components of a quorum sensing signal transduction system . In this report we describe the overexpression and purification of the response regulator, AgrA . Monoclonal antibodies were produced and used to detect AgrA in the cytosolic fraction of S . aureus cells . Purified AgrA did not bind to the RNAIII promoter region in a DNA mobility shift experiment . This confirms previous results obtained with protein extracts from agr+ and agr- cells. J Bacteriol, 1996 Oct, 178(19), 5810 - 2 Identification of LytSR-regulated genes from Staphylococcus aureus; Brunskill EW et al.; In this report, the characterization of a Staphylococcus aureus operon containing two LytSR-regulated genes, lrgA and lrgB, is described . Sequence and mutagenesis studies of these genes suggest that lrgA encodes a murein hydrolase exporter similar to bacteriophage holin proteins while lrgB may encode a protein having murein hydrolase activity. J Invest Dermatol, 1996 Oct, 107(4), 603 - 9 Staphylococcal toxins and protein A differentially induce cytotoxicity and release of tumor necrosis factor-alpha from human keratinocytes; Ezepchuk YV et al.; It has been proposed that toxins and other bacterial protein products of Staphylococcus aureus can act as triggers or persistence factors in several inflammatory skin diseases . In this study, we examined the S . aureus isolates from the skin of patients with atopic dermatitis and psoriasis . We found that the bacterial isolates from these patients exhibited either characteristic superantigenic toxins or thermolabile toxins believed to be staphylococcal alpha-toxin . All of these staphylococcal strains also secreted extracellular staphylococcal protein A . We found significant differences in the action of these toxins on human keratinocytes and keratinocyte cell lines . The superantigenic toxins toxic shock syndrome toxin-1, staphylococcal enterotoxins A and B, and exfoliative toxin-A, as well as staphylococcal protein A, did not induce significant cytotoxic damage in the keratinocyte cell line HaCaT, whereas the staphylococcal alpha-toxin produced profound cytotoxicity . Keratinocyte cytotoxicity induced by staphylococcal alpha-toxin was time and concentration dependent and demonstrated the morphologic and functional characteristics of necrosis, not apoptosis . Addition of alpha-toxin to keratinocytes simultaneously induced cell lysis and tumor necrosis factor-alpha release into the medium within 30 min; apparently, it was constitutive tumor necrosis factor-alpha . On the other hand, superantigenic toxins and, in particular, protein A showed stimulation of tumor necrosis factor-alpha secretion in keratinocytes and release of this cytokine after 6-12 h of incubation . Thus, staphylococcal protein A, alpha-toxin, and superantigenic toxins found in S . aureus isolates from patients with psoriasis and atopic dermatitis can produce direct pro-inflammatory effects on keratinocytes through the release of tumor necrosis factor-alpha . We propose that these effects may be relevant to the induction and persistence of lesions in these two diseases. JAMA, 1996 Sep 25, 276(12), 972 - 7 Infection and allergy incidence in ambulatory surgery patients using white petrolatum vs bacitracin ointment . A randomized controlled trial; Smack DP et al.; OBJECTIVE: To assess the effect of white petrolatum vs bacitracin ointment on wound infection incidence, allergic contact dermatitis incidence, and healing characteristics . DESIGN: Randomized, double-blind, prospective trial comparing white petrolatum with bacitracin ointment in postprocedure wound care . SETTING: A general outpatient dermatology clinic and a tertiary referral advanced surgical procedure clinic at Walter Reed Army Medical Center, Washington, DC . PATIENTS: A total of 922 patients who had dermatologic surgery with a total of 1249 wounds . MAIN OUTCOME MEASURES: The incidence of infection and allergic contact dermatitis during a follow-up period of 4 weeks . Healing characteristics were secondary outcomes . RESULTS: Of the 922 patients enrolled, 440 in the white petrolatum group and 444 in the bacitracin group were evaluable for clinical response . The 2 treatment groups had comparable baseline characteristics . Thirteen patients developed postprocedure infection (1.5%), 9 (2.0%) in the white petrolatum group vs 4 (0.9%) in the bacitracin group (95% confidence interval for difference, -0.4% to 2.7%; P=.37) . Eight infections (1.8%) in the white petrolatum group were due to Staphylococcus aureus vs none in the bacitracin group (P=.004) . No patient in the group using white petrolatum developed allergic contact dermatitis vs 4 patients (0.9%) in the group using bacitracin (P=.12) . Additionally, there were no clinically significant differences in healing between the treatment groups on day 1 (P=.98), day 7 (P=.86), or day 28 (P=.28) after the procedure . CONCLUSIONS: White petrolatum is a safe, effective wound care ointment for ambulatory surgery . In comparison with bacitracin, white petrolatum possesses an equally low infection rate and minimal risk for induction of allergy. Biochem Biophys Res Commun, 1996 Sep 24, 226(3), 730 - 4 Atomic force microscopy proposes a novel model for stem-loop structure that binds a heat shock protein in the Staphylococcus aureus HSP70 operon; Ohta T et al.; The Staphylococcus aureus HSP70 operon produces a polycistronic RNA in response to heat shock, and ORF37 is the first protein to be translated . The promoter of this operon contains a palindromic nucleotide sequence that may form a stem-loop structure . Structural analysis of the promoter regions by atomic force microscopy (AFM) revealed a quadruplet that consists of a pair of stem-loops . A novel "SL2S' (Stem-Loop-Loop-Stem) model was proposed for this structure . AFM also revealed the binding of ORF37 to the quadruplet, establishing a molecular mechanism for this heat shock gene expression; ORF37 acts as a regulator by binding to the SL2S structure in the promoter. Biochemistry, 1996 Sep 24, 35(38), 12251 - 8 Structure and kinetics of the beta-lactamase mutants S70A and K73H from Staphylococcus aureus PC1; Chen CC et al.; Two mutant beta-lactamases from Staphylococcus aureus PC1 which probe key catalytic residues have been produced by site-directed mutagenesis . In the S70A enzyme, the nucleophilic group that attacks the beta-lactam carbonyl carbon atom was eliminated . Consequently, the kcat values for hydrolysis of benzylpenicillin and nitrocefin have been reduced by 10(4)-10(5) compared with the wild-type enzyme . The crystal structure of S70A beta-lactamase has been determined at 2.1 A resolution . With the exception of the mutation site, the structure is identical to that of the native enzyme . The residual activity is attributed either to mistranslation that leads to production of wild-type enzyme and/or to remaining features of the active site that stabilize the tetrahedral transition state . Soaking of the crystals with ampicillin or clavulanate, followed by flash-freezing, has been carried out and the structures examined at 2.0 A resolution . For both experiments, the difference electron density maps revealed buildup of density in the active site that presumably corresponds to beta-lactam binding . However, neither electron density is sufficiently clear for defining the atomic details of the bound compounds . The K73H beta-lactamase has been prepared to test the possible role of Lys73 in proton transfer . It exhibits no detectable activity toward benzylpenicillin, and 10(5)-fold reduction of kcat for nitrocefin hydrolysis compared with the wild-type enzyme . No significant recovery of activity has been measured when the pH was varied between 5.0 and 8.0 . The crystal structure of K73H beta-lactamase has been determined at 1.9 A resolution . While the overall structure is similar to that of the native enzyme, the electrostatic interactions between His73 and neighboring residues indicate that the imidazole ring is positively charged . In addition, the hydroxyl group of Ser70 adopts a position that is incompatible with nucleophilic attack on substrates . A crystal soaked with ampicillin was flash-frozen, and diffraction data were collected at 2.1 A resolution . The electron density map showed no indication of substrate binding. EMBO J, 1996 Sep 16, 15(18), 4789 - 97 Target cell specificity of a bacteriocin molecule: a C-terminal signal directs lysostaphin to the cell wall of Staphylococcus aureus; Baba T et al.; Microbial organisms secrete antibiotics that cause the selective destruction of specific target cells . Although the mode of action is known for many antibiotics, the mechanisms by which these molecules are directed specifically to their target cells hitherto have not been described . Staphylococcus simulans secretes lysostaphin, a bacteriolytic enzyme that cleaves staphylococcal peptidoglycans in general but that is directed specifically to Staphylococcus aureus target cells . The sequence element sufficient for the binding of the bacteriocin as well as of hybrid indicator proteins to the cell wall of S.aureus consisted of 92 C-terminal lysostaphin residues . Targeting to the cell wall of S.aureus occurred either when the hybrid indicator molecules were added externally to the bacteria or when they were synthesized and exported from their cytoplasm by an N-terminal leader peptide . A lysostaphin molecule lacking the C-terminal targeting signal was enzymatically active but had lost its ability to distinguish between S.aureus and S.simulans cells, indicating that this domain functions to confer target cell specificity to the bacteriolytic molecule. Cell Immunol, 1996 Sep 15, 172(2), 205 - 16 Induction of inhibitory activity for B cell differentiation in human CD8 T cells with pokeweed mitogen, dimaprit, and cAMP upregulating agents: countersuppressive effect of platelet factor 4; Crisi GM et al.; As shown previously, native or recombinant (r) human platelet factor 4 (PF4) alleviates the suppression induced by Con A or dimaprit, a histamine type 2 receptor (H2-R) agonist, in a murine system . The effect of rPF4 on human peripheral blood cells has now been studied, using as a model pokeweed mitogen (PWM)-induced, T-cell-mediated suppression of Ig-secreting cell (ISC) formation by Staphylococcus aureus and rIL-2 activated B cells . PWM, but not phytohemagglutinin (PHA), induced inhibitory activity in mitomycin-treated CD8+ T cells, but not unfractionated or CD4+ T cells, for both ISC formation and B cell proliferation . rPF4 and its C-terminal tridecapeptide alleviated the suppressive effect of PWM-activated CD8+ T cells on ISC production but not on proliferation . Heparin did not prevent this immunoregulatory activity of PF4 . Neutralizing antibody to TGF-beta, but not to IFN-gamma or TNF-alpha, alleviated the suppression of ISC formation in some of the experiments . The H2-R appeared to play a part in inducing suppression, because the H2-R antagonist, cimetidine, prevented the PWM-induced suppression of ISC production . Furthermore, dimaprit induced suppression of ISC formation when added instead of PWM at the start of culture . Incubation of CD8+ T cells with dimaprit for only 3 hr prior to coculture with S . aureus + IL-2 activated B cells decreased the ISC response . This suppression was also alleviated by addition of rPF4 to the coculture . Similar to dimaprit, known cAMP upregulating agents, such as forskolin, dibutyryl cAMP, and cAMP analog, all induced this immunoregulatory activity in T cells . Moreover, the effect of dimaprit was prevented by the specific protein kinase A inhibitor, HA1004, suggesting strongly that upregulation of cAMP played a role in the H2-R-mediated effect . Cell contact appeared to be necessary, since supernatants from dimaprit or PWM activated T cells failed to suppress ISC production . We suggest that the known ability of PF4 to prevent TGF-beta-mediated effects on endothelial and other target cells may be involved in the alleviating effect of PF4 on the cell-contact-dependent CD8+ T-cell-mediated B cell suppression. Hosp Pract (Off Ed), 1996 Sep 15, 31(9), 133 - 7, 142-4, 150 Catheter-associated Staphylococcus aureus bacteremia; Gold HS et al.; The majority of cases of Staphylococcus aureus bacteremia are hospital-acquired, and most are associated with infected intravenous catheters . Preventive measures, early detection of infections, and strategies for effective treatment have become matters of increasing urgency. J Immunol, 1996 Sep 15, 157(6), 2555 - 63 L-selectin (CD62L) blockade does not impair peritoneal neutrophil emigration or subcutaneous host defense to bacteria in rabbits; Sharar SR et al.; Neutrophil (PMN) recruitment into systemic inflammatory sites in vivo is thought to be initiated by selectin-mediated endothelial adherence . We explored the role of L-selectin (CD62L) in leukocyte emigration following instillation of bacteria into the peritoneum or s.c . skin in rabbits . Pretreatment with blocking mAb against L-selectin (LAM1.3) reduced peritoneal PMN emigration 4 h after i.p . inoculation with 10(10) CFU of Escherichia coli by only 17% compared with animals receiving a nonblocking L-selectin mAb (LAM1.14) . Peritoneal PMNs from saline-treated rabbits demonstrated a complete absence of L-selectin, whereas those from LAM1.3-treated animals retained 43% of their baseline L-selectin expression . This suggests that L-selectin shedding is not a requisite event for PMN emigration under these conditions . In rabbits given s.c . inoculations with either Staphylococcus aureus or E coli, pretreatment with mAb LAM1.3 did not significantly impair PMN emigration at 24 h, nor increase the incidence, size, or associated mortality of resulting abscesses at 7 days compared with animals receiving nonblocking mAb LAM1.14 . We conclude that: 1) mAb blockade of L-selectin in vivo only modestly affects acute, E . coli-induced peritoneal PMN emigration; and 2) L-selectin blockade does not increase infectious sequelae associated with s.c . bacterial inoculation . These findings of only mildly reduced PMN emigration into the peritoneum and no alteration in s.c . host defense differ from those reported with L-selectin blockade under other, nonbacterial inflammatory conditions, and suggest that redundant selectin-mediated mechanisms (P- and E-selectin) are sufficient for normal PMN emigration in response to bacterial stimulation. J Immunol, 1996 Sep 15, 157(6), 2514 - 20 Bacteria in the bloodstream are trapped in the liver and killed by immigrating neutrophils; Gregory SH et al.; The critical role of the liver in the resolution of systemic bacterial infections is well documented . In the case of Listeria monocytogenes, approximately 60% of bacteria inoculated i.v . into mice are recovered in the liver at 10 min after infection . Here we report that the Listeria recovered at 10 min were distributed equally among the hepatocyte and nonparenchymal liver cell populations . The majority (>/= 75%) of these organisms were bound extracellularly as judged by their sensitivity to gentamicin . In contrast, >/= 93% of Listeria recovered in the liver at 6 h were located within hepatocytes . The listerial burden of the liver decreased 0.5 to 1.0 log, between 10 min and 6 h after infection . This decrease correlated with a sevenfold increase in the percentage of neutrophils that constituted the nonparenchymal cell population . Mice rendered neutrophil deficient by pretreatment with anti-granulocyte (RB6-8C5) mAb exhibited a significant increase (>300%) rather than a decrease in liver Listeria and a marked increase in hepatocyte damage . Similarly, neutrophil-deficient mice exhibited a reduced capacity to eliminate Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus that were cleared by the liver and bound extracellularly to hepatocytes and nonparenchymal cells . These findings document the crucial role of immigrating neutrophils in nonspecific host defenses to systemic bacterial infections expressed within the liver. J Am Vet Med Assoc, 1996 Sep 15, 209(6), 1143 - 6 Evaluation of a coagulase-negative variant of Staphylococcus aureus as a cause of intramammary infections in a herd of dairy cattle; Fox LK et al.; A coagulase-negative variant of Staphylococcus aureus was identified in a herd of 250 lactating dairy cows . During testing of the entire herd, this strain of S aureus was isolated from aseptically collected milk samples of 25 cows . Cows with intramammary infections attributable to coagulase-negative S aureus had an increased somatic cell count in their milk, which was indicative of mastitis infection . Speciation of the Staphylococcus organisms was made, using a series of biochemical tests . A strain of a coagulase-positive S aureus also caused intramammary infections in the herd and shared identical biochemical characteristics with the coagulase-negative strain . Moreover, both strains could not be typed by the use of the International Set of Bovine Phages . Analysis of these findings indicated that a coagulase-negative variant of S aureus can cause intramammary infections in cattle, coagulase-negative variants of S aureus that cause mastitis can be more prevalent in herds than coagulase-positive variants, and clinicians should avoid misclassifying coagulase-negative S aureus as organisms that are clinically unimportant. J Chromatogr A, 1996 Sep 13, 744(1-2), 177 - 85 Comparative peptide mapping of a hepatitis C viral recombinant protein by capillary electrophoresis and matrix-assisted laser desorption time-of-flight mass spectrometry; Winkler MA et al.; Capillary electrophoresis (CE) and matrix assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS) were investigated as alternatives to sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis for peptide mapping with Staphylococcus aureus protease (V8) of a hydrophobic recombinant hepatitis C virus antigen, HC-31, which required 0.1% SDS for solubility . Controls (V8 only) or HC-31 digests were extracted with chloroform-methanol-water (1:4:3) to remove SDS, which interferes with MALDI-TOF, and high salt content, which affects CE . In two different runs by CE, the elution times of each of 11 peptide peaks were very reproducible (R.S.D . < 0.016) . 25 fragments were resolved by MALDI-TOF-MS, including six smaller peptides (M(r) < 13 000) resulting from V8 autodigestion . MALDI-TOF-MS indicated that partial cleavages occurred, primarily at sites where there are paired glutamic and/or aspartic acid residues. J Mol Biol, 1996 Sep 13, 262(1), 21 - 30 Surface display of proteins on bacteriophage lambda heads; Mikawa YG et al.; We have developed plasmid and phage vectors for the display of foreign proteins on the surface of bacteriophage lambda capsid by modifying the D gene which encodes the major head protein gpD . The vectors have multiple cloning sites, and permit colour selection and conditional chain termination for recombinants . Displayed proteins can be fused to either the N or C terminus of gpD by a peptide linker . The conditional chain termination scheme, via a host Escherichia coli suppressor activity, allows the fusion and assembly of homomultimeric proteins as well as control of the number of fusion proteins per phage particle . We have successfully displayed beta-lactamase, IgG-binding domains of the Staphylococcus aureus protein A, and beta-galactosidase by cloning the genes into the vector . The constructs express functionally active proteins fused to gpD that assemble into phage particles . These results suggest that gpD may be fused to many other peptides and proteins at their N or C terminus and the fusion products may be accessible on the surface of bacteriophage lambda particles. Arch Intern Med, 1996 Sep 9, 156(16), 1857 - 60 Microbiology of acute purulent pericarditis . A 12-year experience in a military hospital; Brook I et al.; OBJECTIVE: To study the aerobic and anaerobic micro-biological and clinical characteristics in 15 cases of acute pericarditis treated over a 12-year period . DESIGN: Retrospective review of microbiological and clinical data . SETTING: Military hospital in Bethesda, Md . RESULTS: Aerobic or facultative bacteria alone were present in 7 specimens (47%), anaerobic bacteria alone in 6 specimens (40%), and mixed aerobic-anaerobic flora in 2 specimens (13%) . In total, there were 21 isolates: 10 aerobic or facultative bacteria and 11 anaerobic bacteria, an average of 1.4 per specimen . Anaerobic bacteria predominated in patients with pericarditis who also had mediastinitis that followed esophageal perforation and in patients whose pericarditis was associated with orofacial and dental infections . The predominant aerobic bacteria were Staphylococcus aureus (3 isolates) and Klebsiella pneumoniae (2 isolates), and the predominant anaerobic bacteria were Prevotella species (4 isolates), Peptostreptococcus species (3 isolates), and Propionibacterium acnes (2 isolates) . CONCLUSION: The findings in our study highlight the potential importance of anaerobic bacteria in acute pericarditis. Biochim Biophys Acta, 1996 Sep 6, 1303(1), 63 - 73 Leukotriene B4 and platelet activating factor production in permeabilized human neutrophils: role of cytosolic PLA2 in LTB4 and PAF generation; Bauldry SA et al.; The specific type of phospholipase A2 (PLA2) involved in formation of leukotriene B4 (LTB4) and platelet activating factor (PAF) in inflammatory cells has been controversial . In a recent report we characterized activation of the 'cytosolic' form of PLA2 (cPLA2) in human neutrophils (PMN) permeabilized with Staphylococcus aureus alpha-toxin under conditions where the secretory form of PLA2 (sPLA2) was inactive . In the current study, generation of both LTB4 and PAF in porated PMN are demonstrated . PMN, prelabeled with {3H}arachidonic acid (3H-AA, to assess AA release and LTB4 production) or with 1-O-{9',10'-3H}hexadecyl-2-lyso-glycero-3-phosphocholine (3H-lyso-PAF, for determination of lyso-PAF and PAF formation), were permeabilized with alpha-toxin in a 'cytoplasmic' buffer supplemented with acetyl CoA . Maximum production of both PAF and LTB4 required addition of 500 nM Ca2+, G-protein activation induced with 10 microM GTP gamma S, and stimulation with the chemotactic peptide, N-formyl-Met-Leu-Phe (FMLP, 1 microM); LTB4 production was confirmed by radioimmunoassay . Removal of acetyl CoA from the system had little effect on LTB4 generation but blocked PAF production with a concomitant increase in lyso-PAF formation LTB4 and PAF production occurred in parallel over time and at differing ATP and Ca2+ concentrations . Further work demonstrated that: (i) maximum production of both inflammatory mediators required a hydrolyzable form of ATP; (ii) blocking phosphorylation with staurosporin inhibited production of both; (iii) the reducing agent, dithiotreitol, had little affect on LTB4 formation but slightly enhanced PAF generation . This study clearly shows that cPLA2 activation can provide precursors for both LTB4 and PAF, that maximum PAF and LTB4 formation occur under conditions that induced optimal cPLA2 activation, that a close coupling between LTB4 and PAF formation exists, and that, after substrate generation, no additional requirements are necessary for LTB4 and PAF generation in the permeabilized PMN system. Biochim Biophys Acta, 1996 Sep 5, 1296(2), 228 - 34 C-terminal truncation of spinach chloroplast NAD(P)-depend