Biochem J, 1985 Mar 15, 226(3), 853 - 8 Discontinuities in the topography of alcohol dehydrogenase-sodium dodecyl sulphate complexes revealed by mutagenesis and proteolysis; Ferl RJ; Experiments utilizing proteolytic mapping of maize Alcohol dehydrogenase-1 protein (EC 1.1.1.1; ADH) showed that, in the presence of sodium dodecyl sulphate (SDS), two discrete areas of the protein molecule were hypersensitive to digestion with Staphylococcus aureus V8 proteinase . These areas were mapped to points 11 and 27 kDa along the 41 kDa polypeptide . Furthermore, ADH1 proteins isolated from the ethyl methanesulphonate-induced mutants U725 and W586 showed both a change in electrophoretic mobility in SDS gels, and an altered V8 proteinase digestion pattern . Protein from U725 migrated in SDS gels as though it was 2 kDa smaller than wild-type ADH protein and lacked the 11 kDa cleavage site . Protein from W586 lacked the 30 kDa cleavage site and migrated as if it was 3 kDa smaller than wild type . The possible relationships between proteinase cleavage sites, mutations and SDS gel mobilities are discussed. J Biol Chem, 1985 Mar 10, 260(5), 2670 - 4 The ompA signal peptide directed secretion of Staphylococcal nuclease A by Escherichia coli; Takahara M et al.; The hybrid pre-enzyme formed by fusion of the signal peptide of the OmpA protein, a major outer membrane protein of Escherichia coli, to Staphylococcal nuclease A, a protein secreted by Staphylococcus aureus, is translocated across the cytoplasmic membrane of E . coli with concomitant cleavage of the signal peptide . A DNA fragment containing the coding sequence for the ompA signal peptide was initially ligated to a DNA fragment containing the coding sequence for nuclease A, with a linker sequence of 33 nucleotides separating the coding sequences . When this fused gene was induced, an enzymatically active nuclease was secreted into the periplasmic space; sequential Edman degradation of this protein revealed that the ompA signal peptide was removed at its normal cleavage site resulting in a modified version of the nuclease having 11 extra amino acid residues attached to the amino terminus of nuclease A . The 33 nucleotides between the coding sequences for the ompA signal peptide and the structural gene for nuclease A were subsequently deleted by synthetic oligonucleotide-directed site-specific mutagenesis . The nuclease produced by this hybrid gene was secreted into the periplasmic space and by sequential Edman degradation was identical to nuclease A . Thus, the ompA signal peptide is able to direct the secretion of fused staphylococcal nuclease A, and signal peptide processing occurs at the normal cleavage site . When the hybrid gene is expressed under the control of the lpp promoter, nuclease A is produced to the extent of 10% of the total cellular protein. J Mol Biol, 1985 Mar 5, 182(1), 91 - 107 Analogs of cyclic AMP that elicit the biochemically defined conformational change in catabolite gene activator protein (CAP) but do not stimulate binding to DNA; Ebright RH et al.; We have measured the effects on catabolite gene activator protein (CAP) of 22 synthetic analogs of cAMP . Each analog was assayed to test three parameters: (1) binding to CAP; (2) induction of the conformational change in CAP; and (3) activation of transcription . Thus we have identified seven cAMP analogs that bind to CAP as well or better than does cAMP, cause the assayed conformational change in CAP, yet exhibit no ability to activate transcription . We designate these analogs class D . The conformational change elicited in CAP by the class D analogs was further investigated by: (1) sensitivity to the proteolytic enzymes chymotrypsin, Staphylococcus aureus V8 protease, subtilisin and trypsin; (2) formation of inter-subunit covalent crosslinks by 5,5'-dithiobis(2-nitrobenzoic acid); and (3) degree of labeling of cysteine by {3H}N-ethylmaleimide . These experiments failed to detect a conformational difference between the CAP-class D and CAP-cAMP complexes . Filter binding and nuclease protection experiments indicate that the class D analogs do not efficiently support the binding of CAP to DNA . From these results, we suggest that there exists a hitherto undetected event dependent on cAMP, and required for CAP to bind to DNA . We suggest that this event involves a change that takes place in proximity to the N6 atom of cAMP . Three possible interpretations are discussed. Southeast Asian J Trop Med Public Health, 1985 Mar, 16(1), 15 - 21 Brugia malayi: serum dependent cell-mediated reactions to microfilariae; Chandrashekar R et al.; Sheathed and exsheathed microfilariae of Brugia malayi are killed by normal rat cells in the presence of immune serum in vitro . Immune serum heated at 56 degrees C for 1 hour lost this activity which was largely restored by the addition of fresh normal rat serum . EDTA but not EGTA abolished this activity indicating the operation of complement by alternate pathway . Fresh normal rat serum alone promoted cellular adherence without exerting cytotoxicity to the microfilariae . The activity in the immune serum could be removed with Staphylococcus aureus cells containing Protein A or anti-IgG antiserum . The activity could also be absorbed to and eluted from Protein A--sepharose CL-4B suggesting the involvement of IgG . Neutrophils and macrophages participate in the antibody dependent cell-mediated cytotoxicity phenomenon . Eosinophils while adhering to the microfilariae exert cytotoxicity only to the exsheathed parasites. Ann Inst Pasteur Microbiol, 1985 Mar-Apr, 136A(2), 241 - 5 Gelatin and collagen binding to Staphylococcus aureus strains; Carret G et al.; An interaction between the staphylococcal surface and gelatin is described . Out of 98 Staphylococcus aureus strains, 2 clumped in gelatin solution . Binding of collagen on the Staphylococcus aureus surface was also observed. Res Vet Sci, 1985 Mar, 38(2), 167 - 73 Electron microscopic study of leucocytic infiltration of the mammary teat duct during infection with Staphylococcus aureus; Nickerson SC et al.; Leucocytic response to Staphylococcus aureus infection was observed in the bovine mammary teat duct using transmission electron microscopy . Leucocytes migrating across the stratified squamous epithelium were observed in close association with areas colonised by cocci . Leucocytes gained access across the epithelium to the teat duct lumen by: passage as luminal cells desquamated; migration through degenerate cell cytoplasm; and penetration of cell junctions . The results provide evidence of marked leucocytic infiltration into ductal tissue which may participate in the cellular response to mastitis. Acta Paediatr Scand, 1985 Mar, 74(2), 219 - 25 Defective leukocyte interferon response in children with recurrent infections accompanied by arthralgia; Bondestam M et al.; Children with recurrent and/or unusually severe infections were investigated for possible defects in the interferon (IFN)-natural killer (NK) cell system . Two series, each of 13 children, were examined, one in 1982 and one in 1983 . Healthy children, seven in 1982 and eight in 1983, served as controls . Peripheral blood mononuclear leukocytes were examined for IFN production induced by the IFN-alpha inducers Sendai virus and Escherichia coli and by the IFN-gamma inducers Concanavalin A and Lens culinaris lectin . None of these inducers discriminated patients from controls . However, the bacteria Staphylococcus aureus Cowan I (SACol), inducers of atypical IFN in null lymphocytes, yielded significantly lower IFN production in infection-prone children than in controls, particularly in children with recurrent infections accompanied by arthralgia . No differences in basal NK activity or in the in vitro enhancement of such activity by IFN-alpha were found between patients and controls. Mol Biochem Parasitol, 1985 Mar, 14(3), 275 - 81 In vitro translation of mRNA from Toxocara canis larvae; Sugane K et al.; 300 micrograms of total RNA was extracted from 1 ml of packed Toxocara canis larvae by centrifugation through a 5.7 M cesium chloride cushion . 60 micrograms of polyadenylated messenger RNA was separated from 300 micrograms of total RNA in an oligothymidylic acid-cellulose gel column . The in vitro translation of the mRNA, isolated from T . canis larvae, was carried out using the rabbit reticulocyte cell-free translation system . Incorporation of {35S}methionine into trichloroacetic acid precipitable material in the lysate containing mRNA was 4-5 times greater than that of control . Translation products were analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by autoradiography . Many polypeptides ranging in molecular weight from 10000 to 100000 were synthesised in the lysate . A T . canis positive human serum was mixed with translation products to form antigen-antibody complexes, which were then absorbed by Staphylococcus aureus Cowan 1 strain and analysed by the autoradiography of SDS-PAGE . Three antigenic polypeptides with molecular weights of 49000, 27000 and 22000 which reacted specifically with IgG antibody in T . canis positive human serum, were demonstrated . The 27000 MW polypeptide reacted particularly strongly with the IgG antibody. J Dairy Sci, 1985 Mar, 68(3), 726 - 31 Evaluation of protein A and a commercial bacterin as vaccines against Staphylococcus aureus mastitis by experimental challenge; Pankey JW et al.; Protein A and a commercial staphylococcal bacterin were evaluated by experimental challenge with Staphylococcus aureus (ATCC 29740) . Thirty cows in first lactation were in three treatment groups, protein A, bacterin, and nonvaccinated controls . Studies were through three lactations and included bacteriological and cytological analyses of quarter milk samples . Rate of intramammary infection with Staphylococcus aureus was similar for vaccinated and unvaccinated cows . Rates of spontaneous cure within each lactation were significantly higher for vaccinated cows . For all three lactations, spontaneous cure rates were 83, 73, and 47% for protein A, bacterin, and control cows . Somatic cell counts were significantly lower for vaccinated cows for quarters infected with Staphylococcus aureus, but no differences were demonstrated for milk production by lactation . Incidence of clinical mastitis was higher in unvaccinated cows, but too few developed for a valid comparison. Clin Rheumatol, 1985 Mar, 4(1), 90 - 2 Staphylococcus aureus infection complicating haemarthroses in elderly patients; Helliwell M; The case histories of 3 elderly patients who developed haemarthrosis of osteoarthritic joints with subsequent infection with staphylococcus aureus, are described . Trauma to the affected joints was a predisposing factor in 2 patients and, while only one patient developed clinical signs of sepsis, all had marked elevation of the erythrocyte sedimentation rate . Although suspicion of joint sepsis was obscured by the presence of a coexistent haemarthrosis, routine culture of joint aspirates showed infection with staphylococcus aureus and all patients recovered well with antibacterial therapy. Can J Biochem Cell Biol, 1985 Mar, 63(3), 195 - 203 The cross-linking of human Met-hemoglobin with {14C}dimethyl adipimidate; Ferris RJ et al.; Met-hemoglobin, cross-linked with {14C}dimethyladipimidate (cross-linking span 9 A; 1 A = 0.1 nm), yields four distinct molecular weight products (monomer, dimer, trimer, and tetramer) as observed on sodium dodecyl sulfate - polyacrylamide gels . The dimer species was purified to homogeneity by gel filtration . It was proteolytically degraded using a combination of Staphylococcus aureus V8 protease, pepsin, trypsin, and chymotrypsin . The resultant peptides were fractionated using gel filtration and ion-exchange chromatography methods . Two cross-links were unambiguously identified: (i) alpha 1Lys7-alpha 1Lys11 and (ii) beta 1Lys82-beta 2Lys82 . The identified cross-links correlated well with the known structure of hemoglobin . However, attempts to isolate and identify a greater number of cross-linked peptides were unsuccessful owing to the complexity of the peptide mixtures . The complexity was a direct result of the chemical modification of the hemoglobin molecule . Therefore, attempts to employ chemical cross-linking as a means of examining sites of protomer contact within large oligomeric proteins should be approached with caution. J Bone Joint Surg Br, 1985 Mar, 67(2), 229 - 31 Infection in experimental hip arthroplasties; Southwood RT et al.; The relationship between the route of inoculation, the dose of inoculum and the development of infection after prosthetic replacement has been determined in an animal model . The rabbit hip served as the model and a Staphylococcus aureus isolated from an infected human hip arthroplasty was introduced using different protocols . The 50% infective dose (ID50) was determined for comparative purposes . Contamination of the wound site with only a few bacteria was likely to result in infection . It was considerably more difficult to induce infection when the operation was performed without inserting the prosthesis, which suggests that the implant inhibits the body's mechanism for dealing with the insult . It was difficult to produce infection by inoculating the organisms into the bloodstream: if this inoculation was delayed till three weeks after operation the animals were often grossly septicaemic by the time the arthroplasty was infected . The results emphasise the importance of minimising intra-operative contamination and the efficacy of antibiotic cover. Plast Reconstr Surg, 1985 Mar, 75(3), 394 - 6 Cellular and bacterial toxicities of topical antimicrobials; Lineaweaver W et al.; Cellular and bacterial toxicities of four commonly used topical antimicrobials (1% povidone-iodine, 3% hydrogen peroxide, 0.25% acetic acid, and 0.5% sodium hypochlorite) were assayed in vitro using cultures of human fibroblasts and Staphylococcus aureus . All agents tested at full strength killed 100 percent of exposed fibroblasts . Fibroblast toxicity exceeded bacterial toxicity with serial dilutions of hydrogen peroxide and acetic acid . Dilutions of povidone-iodine (1:1000) and sodium hypochlorite (1:100) were identified where no fibroblast toxicity occurred while full bactericidal activity persisted. J Am Geriatr Soc, 1985 Mar, 33(3), 170 - 4 Septic arthritis in the elderly; McGuire NM et al.; The clinical and microbiologic features of septic arthritis in 23 elderly patients are reviewed . Fifteen patients had pre-existing joint diseases, predominantly osteoarthritis and rheumatoid arthritis . Eight patients had underlying systemic illnesses, and eight patients were receiving systemic corticosteroid therapy prior to the development of septic arthritis . The knee was the joint most commonly infected . Although Staphylococcus aureus was the major pathogen (52.2 per cent of patients), enteric gram-negative bacilli were found in seven of 23 patients (30.4 per cent) . Five patients died (21.7 per cent mortality), two as a result of their infection and three of nosocomial Pseudomonas sepsis . Eight of the 18 survivors (44.4 per cent) developed osteomyelitis in the contiguous bone . Return of joint function was slow in all patients . Septic arthritis in the elderly is difficult to treat and has a poor outcome, possibly because pre-existing joint disease is very common and enteric gram-negative bacilli are often the causative organisms. Infect Immun, 1985 Mar, 47(3), 710 - 2 Effect of toxic shock syndrome toxin 1 on chicken embryos; de Azavedo JC et al.; Staphylococcus aureus strains associated with toxic shock syndrome produce toxic shock syndrome toxin 1 (TSST1) . This toxin has a variety of biological effects, including enhanced lethality in rabbits in the presence of sublethal amounts of lipopolysaccharide (LPS) . Because chicken embryos are highly susceptible to LPS, the synergistic effect of TSST1 and LPS was examined in this system . Although TSST1 per se had no effect on chicken embryos, it potentiated the lethal effect of LPS . The 50% lethal dose of LPS was greatly reduced in the presence of up to 10 micrograms of TSST1 per ml . However, at high doses of TSST1 (greater than 100 micrograms/ml), no enhanced lethality was observed . The lowest dose of TSST1 tested which potentiated lethality was 10 ng/ml. Infect Immun, 1985 Mar, 47(3), 581 - 6 Model of experimental chronic osteomyelitis in rats; Rissing JP et al.; We describe here a Sprague-Dawley rat model for chronic osteomyelitis . Staphylococcus aureus and sodium morrhuate were implanted by either microdrilling or direct needle injection into the tibiae of rats . Of 107 rats, 87 (81%) developed osteomyelitis when a high-speed drill was used for implantation, and 27 (51%) of 53 rats developed osteomyelitis by direct needle inoculation (chi square = 9.81, P less than 0.01) . Demonstrated histopathological changes included the presence of resorption bays filled with osteoclasts . Quantitative microbiological monitoring of tibial count confirmed disease chronicity, yielding stable numbers of CFU (10(6.29 +/- 0.27) ) of S . aureus over 70 days . Infected animals became anemic and lost weight . The erythrocyte sedimentation rates and leukocyte counts were not elevated . Roentgenograms provided the best correlation with the number of organisms in infected tibiae (r2 = 0.80) . Rats with infected tibiae were treated with either oxacillin (120 mg/kg per day) or ceftriaxone (50 mg/kg per day) . Treatment over 14 or 28 days reduced S . aureus counts in tibiae but did not reliably sterilize infected bones, suggesting that this model was resistant to prolonged antimicrobial therapy. Cancer Res, 1985 Mar, 45(3), 1263 - 9 Effects of plasma treatment with purified protein A and Staphylococcus aureus Cowan I on spontaneous animal neoplasms; Klausner JS et al.; Eleven dogs with spontaneous neoplasms were intensively treated with an immunoadsorption system consisting of a continuous flow centrifuge, Protein A-Sepharose columns, and a semi-automatic elution system . Despite consistent and substantial lowering of immunoglobulin G levels, tumor regression was noted in only one of 11 dogs . In contrast, infusion of small volumes of plasma after incubation with heat and formalin-treated Staphylococcus aureus Cowan I resulted in a tumoricidal response in five of six animals . These results suggest that tumor necrosis is probably not induced by Protein A-mediated removal of humoral "blocking" factors. J Antimicrob Chemother, 1985 Mar, 15(3), 353 - 63 Ceftazidime plus mezlocillin as initial antibiotic therapy in febrile neutropenic patients with haematological malignancy; Clough JV et al.; Sixty episodes of fever in neutropenic patients with haematological malignancy were treated with ceftazidime and mezlocillin . Improvement or temporary improvement was seen in 76% of patients with microbiologically or clinically documented infection . Fifty-seven per cent of episodes due to bacteraemia improved with the antibiotics . Escherichia coli and Staphylococcus aureus were the commonest pathogens isolated; bacteraemia due to Staph . epidermis was not encountered . The toxicity of ceftazidime plus mezlocillin was acceptable . Diarrhoea developed in 12% and a skin rash in 10%. J Exp Med, 1985 Mar 1, 161(3), 490 - 502 Human interleukin 1 . Purification to homogeneity; Kronheim SR et al.; We have purified human interleukin 1 (IL-1) to homogeneity by a simplified procedure that results in excellent yields of pure material that retains a high level of biological activity . IL-1, secreted by human peripheral blood macrophages that have been stimulated with Staphylococcus aureus, was purified by ion exchange chromatography and affinity chromatography on Procion Red agarose . The pure protein has a specific activity of 3.2 X 10(8) U/mg in the thymocyte mitogenesis assay, and is pyrogenic . No molecular weight heterogeneity was observed, in contrast to findings for mouse IL-1 and earlier reports of human IL-1 . Purified IL-1, as analyzed by two-dimensional electrophoresis/electrofocusing gels, exhibited a series of charged species with isoelectric points ranging from 6.0 to 4.9, all with a molecular weight of approximately 17,500 . Amino acid analysis indicated an abundance of acidic residues, in agreement with the low isoelectric points . There is little or no cysteine in the molecule . No evidence was found for the presence of carbohydrate moieties . The overall yield for this procedure was approximately 31% of the activity contained in the initial culture supernatant. J Infect Dis, 1985 Mar, 151(3), 514 - 22 Induction of human interleukin-1 by toxic-shock-syndrome toxin-1; Parsonnet J et al.; Strains of Staphylococcus aureus isolated from patients with toxic shock syndrome (TSS) make a characteristic protein known as toxic-shock-syndrome toxin-1 (TSST-1), but the role of this protein in the pathogenesis of TSS is not certain . We have purified TSST-1 by using a combination of alcohol precipitation, isoelectric focusing, and gel chromatography . TSST-1 has an isoelectric point of 7.2 and a molecular weight of 23,100, in accordance with previously published determinations for this protein, and is serologically identical to pyrogenic exotoxin C and staphylococcal enterotoxin F . In highly purified form, TSST-1 is a potent inducer of interleukin-1 production by human monocytes, as quantitated in a thymocyte-proliferation assay . This capability is not attributable to contamination by other staphylococcal products or gram-negative endotoxin and can be blocked by hydrocortisone . Many features of TSS suggest that induction of interleukin-1 by TSST-1 in vivo may play a central role in the elaboration of this disease. J Immunol, 1985 Mar, 134(3), 1690 - 701 The roles of T cell factors in activation, cell cycle progression, and differentiation of human B cells; Jelinek DF et al.; The relationship of the T cell influences involved in human B cell activation and differentiation into immunoglobulin-secreting cells (ISC) was investigated . T cell supernatants (T supt) generated by stimulating T cells with phytohemagglutinin and phorbol myristate acetate contained activities capable of augmenting DNA synthesis and the growth of mitogen-stimulated B cells and supporting the differentiation of ISC . To examine the role of T supt in B cell activation and the progression through the cell cycle, T cell- and monocyte-depleted B cells were stimulated with formalinized Cowan I strain Staphylococcus aureus (SA), and the percentages of cells in G1, S, and G2 + M were determined by acridine orange staining and analysis . In all experiments, a similar percentage of cells entered G1 during the first 24 to 36 hr of culture when stimulated with SA or SA + T supt . Similar results were seen when B cell activation was analyzed by acquisition of a number of other markers of cell activation . Analysis of cell cycle progression with mithramycin staining of cellular DNA in the presence or absence of vinblastine to arrest mitosis indicated that SA-activated B cells were able to complete S and divide in the absence of T supt . Although an effect of T supt on the progression of B cells through the S phase was evident during the first cell cycle, the major effect only became apparent after the first round of cell division . Although T supt was not necessary for initial B cell activation, T cell influences were absolutely necessary for the differentiation of ISC . T supt did not need to be present during the initial 24 to 36 hr of incubation to permit subsequent generation of ISC . However, when T supt was present initially, an increased number of ISC were produced . Hydroxyurea elimination of cells traversing the G1-S interphase indicated that reception of the differentiation signal occurred before the S phase, but that the generation of ISC required subsequent DNA synthesis and/or cell division . Although precursors of ISC were entirely contained within the population triggered to divide by SA alone, there was no preferential expansion of such precursors as a result of SA stimulation . These results indicate that T cell signals are not absolutely necessary for initial B cell activation and progression through the first cell cycle, although T cell factors promote DNA synthesis by some activated B cells . In contrast, differentiation into ISC is completely dependent on T cell influences.(ABSTRACT TRUNCATED AT 400 WORDS) J Antimicrob Chemother, 1985 Mar, 15(3), 291 - 5 The in-vitro activity of some antimicrobial agents against methicillin-resistant Staphylococcus aureus; Moorhouse EC et al.; A total of 185 strains of methicillin resistant Staphylococcus aureus was investigated for sensitivity to five other antimicrobial agents . The vast majority of these strains were also resistant to gentamicin and fusidic acid . Rifampicin was the most active drug tested (MIC90, 0.007 mg/l), while two newer compounds teichomycin and ciprofloxacin showed equal and appreciable activity (MIC90, 0.5 mg/l). Infect Immun, 1985 Mar, 47(3), 598 - 604 Acquired ability of Staphylococcus aureus to produce toxic shock-associated protein and resulting illness in a rabbit model; Rasheed JK et al.; Staphylococcus aureus from patients with toxic shock syndrome (TSS) produce TSS toxin 1 . We transferred, by a bacteriophage, the ability to produce TSS toxin 1 from a TSS toxin 1-positive to a TSS toxin 1-negative strain of S . aureus . This recombinant strain produced TSS toxin 1 as confirmed by isoelectric focusing, immunodiffusion, radioimmunoassay, and autoradiography . The recombinant produced TSS-like illness in rabbits, and was significantly (P less than 0.001) more lethal than the recipient strain . Both strains produced fever and diarrhea, but, in addition, rabbits challenged with the recombinant also developed lowered blood pressure (P = 0.002), conjunctival hyperemia, erythroderma, and respiratory distress . Histopathological findings in rabbits challenged with the recombinant strain were remarkably similar to those described for humans with TSS, e.g., erythrophagocytosis, liver "triaditis," and vasodilatation . This study demonstrates that this protein may contribute to the pathogenesis of the TSS. J Immunol, 1985 Mar, 134(3), 1397 - 402 Graft-vs-host reactions (GVHR) across minor murine histocompatibility barriers . I . Impairment of mitogen responses and suppressor phenomena; Holda JH et al.; In our laboratory, we have developed a murine model to examine GVHD across minor histocompatibility antigens . In our model, GVHD is induced by injecting B10.D2 spleen cells into irradiated BALB/c recipients . Seven to 10 days after irradiation and injection of cells, there are significant changes in cell function in the recipient spleens . In the B10.D2----BALB/c (600 rad) model, recipient spleen cells are profoundly unresponsive to Con A and LPS stimulation but show increased B cell activity measured by Staphylococcus aureus protein A plaque-forming activity . Spleen cells from such GVH mice profoundly suppress the mitogenic responses of normal BALB/c or B10.D2 spleen cells to Con A and LPS . The degree of impairment of the mitogenic response and the ability to suppress normal cells is proportional to the dose of cells used to induce GVH reactions . Both the inability to respond to mitogens and the capacity to suppress are also related to the dose of irradiation given to the recipients . In addition, immunosuppression across minor histocompatibility antigens shows an unevenhandedness . If we inject parental B10.D2 or BALB/c cells into F1 recipients (P----F1), there is greater inhibition of mitogenic responses when B10.D2 parental cells are given than when BALB/c cells are given to the irradiated F1 recipients . These experiments show that significant immunosuppression occurs during GVH reactions across minor histocompatibility barriers . The degree of suppression varies according to the dose of cells used to induce GVH, the dose of irradiation to the recipient and the "strength" of the GVH recognition system . Such experiments provide models for GVH disease seen in humans who receive treatment for leukemia or other diseases that involves recipient irradiation and infusion of HLA-identical bone marrow. Biochem J, 1985 Mar 1, 226(2), 369 - 77 Physicochemical behaviour and structural characteristics of membrane-bound acetylcholinesterase from Torpedo electric organ . Effect of phosphatidylinositol-specific phospholipase C; Futerman AH et al.; Quantitative solubilization of the phospholipid-associated form of acetylcholinesterase (AChE) from Torpedo electric organ can be achieved in the absence of detergent by treatment with phosphatidylinositol-specific phospholipase C (PIPLC) from Staphylococcus aureus {Futerman, Low & Silman (1983) Neurosci . Lett . 40, 85-89} . The sedimentation coefficient on sucrose gradients of AChE solubilized in detergents (DSAChE) varies with the detergent employed . However, the coefficient of AChE directly solubilized by PIPLC is not changed by detergents . Furthermore, PIPLC can abolish the detergent-sensitivity of the sedimentation coefficient of DSAChE purified by affinity chromatography, suggesting that one or more molecules of phosphatidylinositol (PI) are co-solubilized with DSAChE and remain attached throughout purification . DSAChE binds to phospholipid liposomes, whereas PIPLC-solubilized AChE and DSAChE treated with PIPLC do not bind even to liposomes containing PI . Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis shows that PIPLC-solubilized AChE, like unmodified DSAChE, is a catalytic subunit dimer; electrophoresis in the presence of reducing agent reveals no detectable difference in the Mr of the catalytic subunit of unmodified DSAChE, of AChE solubilized by PIPLC and of AChE solubilized by Proteinase K . The results presented suggest that DSAChE is anchored to the plasma membrane by one or more PI molecules which are tightly attached to a short amino acid sequence at one end of the catalytic subunit polypeptide. Am J Pathol, 1985 Mar, 118(3), 367 - 78 Clinicopathologic responses in cats with feline leukemia virus-associated leukemia-lymphoma treated with staphylococcal protein A; Engelman RW et al.; Purified protein A from Staphylococcus aureus Cowan I was injected intraperitoneally or was incorporated in filters ex vivo through which plasma from cats with feline leukemia virus (FeLV)-associated leukemia-lymphoma was passed . Before treatment, 65% of the FeLV-infected cats were anemic, and 70% were thrombocytopenic . Concomitant infections, or immune-mediated disease, was common . During treatment 50% of the cats with FeLV-associated disease improved objectively with normal posttreatment hematocrits, thrombocyte and leukocyte counts, disappearance of dysplastic hematologic elements, and correction of marrow dyscrasias . A 33% response to treatment occurred in cats with unequivocal manifestations of malignant disease and was characterized by reductions in tumor size and marrow and peripheral blood neoplastic cell populations . Clearance of FeLV viremia was documented in 28% of the treated cats . The several possible mechanisms by which treatment with staphylococcal protein A causes reduction in the extent of malignant disease are considered. J Hosp Infect, 1985 Mar, 6 Suppl A, 33 - 6 Disinfectant properties of new povidone-iodine preparations; Schubert R; Quantitative suspension tests performed with Staphylococcus aureus ATCC 6538 and different preparations of povidone-iodine (PVP-I) show that the germicidal activity of a preparation depends on the content of free, non-complex-bound iodine . A comparison between the results obtained with different PVP-I preparations showed that other substances present influenced microbicidal activity. J Hosp Infect, 1985 Mar, 6 Suppl A, 195 - 9 The management of methicillin-resistant Staphylococcus aureus in a major hospital; Tyzack R; A reduction in the incidence and duration of methicillin-resistant Staphylococcus aureus infection and colonization was obtained by the introduction of a rigorous control programme . This included computerization of data, improved nursing practices and an antiseptic routine. J Immunol, 1985 Mar, 134(3), 1609 - 18 Monoclonal antibodies to murine gamma-interferon which differentially modulate macrophage activation and antiviral activity; Schreiber RD et al.; Four monoclonal IgG antibodies to purified, recombinant murine gamma-interferon (rIFN-gamma) have been produced by fusion of immune hamster splenocytes with HAT-sensitive murine myeloma cells . Specificity was confirmed either with an enzyme-linked immunosorbent assay (ELISA) that used immobilized rIFN-gamma or with a radioimmunoassay that employed soluble 125I-rIFN-gamma and heat-killed, fixed Staphylococcus aureus-bearing Protein A . Competition binding experiments suggested that the monoclonal antibodies (MoAb) displayed two distinct epitope specificities: one displayed by H1 and H2, and the other displayed by H21 and H22 . By using murine-human recombinant IFN-gamma hybrid molecules, the H1/H2 epitope was shown to depend on the amino-terminus of IFN-gamma, whereas the H21/H22 epitope was formed by the carboxy-terminal amino acid sequence . The MoAb also reacted with natural IFN-gamma . When bound to a surface, all four MoAb, but not normal hamster IgG, removed 100% of the antiviral and MAF activities present in supernatants of cultures of the murine 24/G1 T cell hybridoma . In free solution, all four antibodies inhibited IFN-gamma dependent antiviral activity, but with different efficiencies . Soluble H21/H22 also blocked all of the 24/G1-derived activity that induces nonspecific tumoricidal activity in macrophages (MAF) while H1/H2 enhanced MAF activity . The differential inhibitory or enhancing activities of H21 or H1 reflected their ability to inhibit or enhance binding of 125I-rIFN-gamma to macrophages, respectively . Soluble H21/H22 and solid-phase H1/H2 inhibited 100% of the MAF, microbicidal, and Ia-inducing activities from lymphokine preparations produced by mitogen stimulation of normal murine splenic cells . These results help to establish definitive structure-function relationships for the IFN-gamma molecule, and indicate that IFN-gamma is the primary lymphokine responsible for inducing nonspecific tumoricidal activity and Ia antigen expression, and for enhancing microbicidal activity in macrophages. Coll Relat Res, 1985 Mar, 5(2), 181 - 91 Monoclonal antibodies against chick gp 115, a matrix glycoprotein with broad distribution; Colombatti A et al.; Hybridoma cell lines were generated producing monoclonal antibodies to chick gp 115, a 115,000-dalton glycoprotein widely distributed in the connective tissue . The specificity of the antibodies was determined by indirect radioimmunobinding: the extent of binding was a function of i) antigen and ii) antibody concentration; iii) inhibition of binding of radiolabelled antibody by unlabelled antibody and iv) among many known extracellular collagenous or noncollagenous glycoproteins tested only gp 115 gave a strong positive binding reaction . The antibodies were used for indirect immunofluorescence and a strong staining reaction was detected in all blood vessels, around smooth muscle cells in several organs, and in the connective matrix of other tissues such as the liver, and the lung . Based on the competition of binding of {125I}-labeled purified antibody by unlabeled antibodies, two separate epitopes were identified on gp 115 . Further analysis of the localization of the epitope was obtained by CNBr cleavage and partial digestion of gp 115 with Staphylococcus aureus V8 protease and alpha-chymotrypsin digestion . Following CNBr cleavage a major fragment of Mr = 35,000 was recognized by 4 monoclonal antibodies, and fragments of comparable Mr were detected following V8 protease and alpha-chymotrypsin digestion. Biochemistry, 1985 Feb 26, 24(5), 1164 - 8 Amino acid sequence of the amphiphilic phosphocarrier protein factor IIILac of the lactose-specific phosphotransferase system of Staphylococcus; Stuber K et al.; The lactose-specific factor III of the phosphotransferase system of Staphylococcus aureus is an amphiphilic trimeric protein composed of identical subunits . It is hydrophilic in its unphosphorylated state and can be isolated from the cytoplasmic protein fraction . It becomes a constituent of the membrane-bound phosphotransferase complex upon phosphorylation of a single histidyl residue . The sequence of S . aureus factor IIILac was determined and revealed that the subunits consist of 103 residues corresponding to a Mr of 11 367 and of 34 101 for the native trimer: (sequence; see text) According to this sequence and previous work histidine residue 82 located in the C-terminal part of the polypeptide chain is phosphorylated at the N-3 position by phosphoenolpyruvate, enzyme I, and histidine-containing phosphocarrier protein . The N-terminal part of the protein comprising approximately one-third of the chain exhibits in vitro affinity toward membrane-bound enzyme IILac. J Biol Chem, 1985 Feb 25, 260(4), 2345 - 54 Human tumor necrosis factor . Production, purification, and characterization; Aggarwal BB et al.; Human tumor necrosis factor (TNF) was purified to homogeneity from serum-free tissue culture supernatants of the HL-60 promyelocytic leukemia cell line induced by 4 beta-phorbol 12-myristate 13-acetate . The purification scheme consisted of controlled-pore glass and DEAE-cellulose chromatography, Mono Q-fast-protein liquid chromatography, and reverse-phase high performance liquid chromatography . The purified protein was homogeneous by the criteria of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and NH2-terminal sequence analysis . The specific activity of purified tumor necrosis factor is approximately 10(8) units/mg . The protein has a molecular weight of approximately 17,000, an isoelectric point of 5.3, and contains two cysteines involved in a disulfide bridge . Approximately 50% homology between TNF and another cytolytic lymphokine, lymphotoxin, exists when the NH2-terminal 34 residues of TNF and internal sequence generated by tryptic, Staphylococcus aureus V8 protease, and chymotryptic digests of TNF are aligned with the complete amino acid sequence of lymphotoxin. Anal Biochem, 1985 Feb 15, 145(1), 27 - 36 Separation of complexes containing protein A and IgG or Fc gamma fragments by high-performance liquid chromatography; Das C et al.; Protein A of Staphylococcus aureus is a bivalent Fc receptor that can form complexes with immunoglobulin G (IgG) or Fc gamma fragments that activate humoral (e.g., complement) and cellular (e.g., lymphocyte) components of the immune system both in vitro and in vivo . To obtain complexes formed between protein A of Staphylococcus aureus (SpA) and rabbit IgG or Fc gamma fragments for purposes of characterizing their compositions and studying their biological activities, we have used high-performance liquid chromatography to separate complexes in 20 min . Complexes were prepared with trace amounts of 125I-SpA and 131I-IgG or 131I-Fc gamma to simplify the analyses . With excess molar amounts of IgG or Fc gamma the complexes have the molecular formulas {(IgG)2SpA}2 or {(Fc gamma)2SpA}2 . With excess SpA, complexes corresponding to (IgG)(SpA) or (Fc gamma)(SpA) are formed, perhaps with other complexes that have different ratios of components . Since SpA is a rod-shaped molecule it elutes at a molecular weight corresponding to 240,000 rather than the true value of 42,000 . This behavior is reflected in the elution of certain complexes at shorter retention times than expected on the basis of actual molecular weights, and facilitates separation of complexes from free IgG or Fc gamma . The true molecular weights and molecular formulas of complexes isolated by HPLC were verified by ultracentrifugation . This HPLC method was used to study the interconversion and stability of complexes. Biochemistry, 1985 Feb 12, 24(4), 1029 - 35 One-step immunoaffinity purification of active progesterone receptor . Further evidence in favor of the existence of a single steroid binding subunit; Logeat F et al.; A very high capacity immunoaffinity matrix for the purification of progesterone receptor was prepared by cross-linking a monoclonal antireceptor antibody to protein A-Sepharose through the Fc fragment . The monoclonal antibody was selected for its property of losing affinity for the receptor at pH 10.5, i.e., in conditions where the receptor remains stable for extensive periods of time . This made it possible to elute active receptor form the immunosorbent . From crude rabbit uterine cytosol the steroid-receptor complexes were purified in a single step . A 1-mL column (containing 7 mg of monoclonal antibody) bound 1600 pmol of steroid-receptor complexes of which 79.5% were eluted . The overall yield of purification was 49% . The specific activity of the purified steroid-receptor complexes was 6.71 +/- 0.79 nmol of bound steroid/mg of protein (mean +/- SE of four experiments) . The purified receptor consisted of a mixture of 110 000- and 79 000-dalton forms . The latter appeared to be produced by proteolysis of the larger form during purification since immunoblot experiments showed that, at the start of purification, the 110 000-dalton form was present in overwhelming majority (80-95%) in the uterine cytosol and that the 79 000-dalton form only appeared during purification . This conclusion was also supported by the peptide analysis of both forms of receptor: the purified receptor was denatured and labeled with 125I; the 110 000- and 79 000-dalton forms were isolated by gel electrophoresis in denaturing conditions and electroelution and were then submitted to mild or extensive digestions by trypsin, chymotrypsin, and protease V8 from Staphylococcus aureus.(ABSTRACT TRUNCATED AT 250 WORDS) Biochemistry, 1985 Feb 12, 24(4), 903 - 10 Mechanism of inhibition of the PC1 beta-lactamase of Staphylococcus aureus by cephalosporins: importance of the 3'-leaving group; Faraci WS et al.; The hydrolysis of cephalosporins containing good leaving groups at the 3'-position {those used in this study were the chromogenic cephalosporin PADAC {pyridine-2-azo-4'-(N',N'-dimethylaniline) substituted on cephalosporin}, cephaloridine, and cephalothin}, catalyzed by the Staphylococcus aureus PC1 beta-lactamase, proceeds in two spectrophotometrically observable phases . The first involves formation of an acyl-enzyme intermediate while the second involves partitioning of this intermediate between two pathways . One path yields the normal cephalosporoate (3) from which the 3'-leaving group is spontaneously eliminated in solution to give the 3-methylenedihydrothiazine 2, while the second involves initial elimination of the 3' substituent, thus yielding a second acyl-enzyme intermediate, which then hydrolyzes to give the same final product as from the first pathway . The second acyl-enzyme is relatively inert to hydrolysis (t1/2 congruent to 10 min at 20 degrees C), and its formation thus leads to transient inhibition of the enzyme . The partition ratio between hydrolysis and elimination at the enzyme active site could be determined either spectrophotometrically from burst experiments or from measurements of residual beta-lactamase activity as a function of cephalosporin concentration . This ratio varied with the leaving group ability of the 3' substituent (acetoxy greater than N,N-dimethylaniline-4-azo-2'-pyridinium greater than pyridinium) in the anticipated fashion . The inert acyl-enzyme intermediate was isolated by exclusion chromatography and shown to contain the cephem nucleus, but not the 3' substituent, covalently bound to the enzyme . As would be expected, PADAC, cephaloridine, and cephalothin yielded the same inert intermediate . Cephalosporins with poor or no 3'-leaving groups, e.g., dansylcephalothin and desacetoxycephalothin, neither displayed the branched pathway nor yielded the long-lived acyl-enzyme. J Biol Chem, 1985 Feb 10, 260(3), 1661 - 9 Phospholipid protection against proteolysis of D-beta-hydroxybutyrate dehydrogenase, a lecithin-requiring enzyme; Maurer A et al.; D-beta-Hydroxybutyrate dehydrogenase is a lipid-requiring enzyme which is localized on the inner face of the mitochondrial inner membrane . The apodehydrogenase, i.e . the purified enzyme devoid of lipid, has been purified from beef heart mitochondria and as such is inactive . It can be reactivated by insertion into phospholipid vesicles containing lecithin . Proteolytic digestion with different proteases has been carried out to obtain insight into the orientation of the enzyme in the membrane and to assess the extent of immersion of the protein into the phospholipid bilayer . Digestion of the apodehydrogenase with either trypsin, chymotrypsin, Staphylococcus aureus protease, thermolysin, carboxypeptidases A and Y, or Pronase (from Streptomyces griseus) leads to loss of activity, as assayed with phospholipid . Limited digestion with carboxypeptidase results in complete inactivation . Of the proteases tested, only Pronase and chymotrypsin cleave and inactivate the enzyme inserted into phospholipid vesicles (enzyme-phospholipid complex) . For the enzyme-phospholipid complex, the loss of activity with Pronase digestion follows a single exponential decay to less than 10% of the initial activity . With chymotrypsin digestion, the staining intensity of the original approximately 31,500-dalton polypeptide decreases more rapidly than the loss of enzymic activity . The enzyme-phospholipid complex, after limited cleavage with chymotrypsin, retains enzymic activity and resonance energy transfer from protein to bound NADH and an approximately 26,000-dalton polypeptide is observed . Phospholipid alters the cleavage pattern with both chymotrypsin and Pronase, and the rate of inactivation of the enzyme-phospholipid complex is slowed in the presence of NAD(H) . Moreover, the rate of inactivation of the apodehydrogenase with chymotrypsin is diminished approximately 3-fold in the presence of NAD+ . Digestion of submitochondrial vesicles with either trypsin, chymotrypsin, or Pronase rapidly inactivates D-beta-hydroxybutyrate dehydrogenase; the addition of NAD+ or NADH, together with dithiothreitol and increased salt (to 50 mM), decreases the rate of inactivation, and with trypsin, virtually eliminates inactivation.(ABSTRACT TRUNCATED AT 400 WORDS) Biochim Biophys Acta, 1985 Feb 4, 827(2), 127 - 34 Purification and structural comparisons of the cytosolic and mitochondrial isoenzymes of fumarase from pig liver; O'Hare MC et al.; A method has been developed for the purification of cytosolic and mitochondrial isoenzymes of fumarase from total homogenates of pig liver . Separation of the isoenzymes from one another was achieved using chromatofocusing . The isoenzymes were pure as judged by production of single bands on electrophoresis in the presence of sodium dodecyl sulphate; they appeared to have identical or very similar subunit molecular weights . The isoenzymes differed in electrophoretic properties under nondenaturing conditions . One-dimensional peptide maps of fragments produced from the two isoenzymes by chemical cleavage at cysteine residues were identical; maps obtained after digestion with the V8 proteinase from Staphylococcus aureus showed small differences at short times of digestion which could have reflected variations in rates of hydrolysis rather than structural differences . However, two-dimensional peptide maps of digests obtained by treatment of the isoenzymes with trypsin followed by chymotrypsin had 58 peptides in common, but showed two peptides unique to the mitochondrial isoenzyme and five peptides unique to the cytosolic form . Using the dansylation procedure, the mitochondrial isoenzyme was shown to have N-terminal alanine and the cytosolic form to have N-terminal glutamic acid or glutamine . We conclude that the isoenzymes of fumarase are identical over nearly all of their amino acid sequences but differ at their N-termini; the extent of these differences is yet to be established . These results are consistent with the claim (Edwards, Y.H . and Hopkinson,D.A . (1979) Ann . Human Genet . Lond . 42, 303-313) that the isoenzymes are determined at the same genetic locus, but they raise interesting questions about the biosynthesis of the isoenzymes. Mol Biochem Parasitol, 1985 Feb, 14(2), 127 - 39 Biosynthesis of two stage-specific membrane proteins during transformation of Plasmodium gallinaceum zygotes into ookinetes; Kumar N et al.; We have studied the synthesis and expression of surface proteins in zygotes of Plasmodium gallinaceum during their transformation to mature ookinetes . The cells were biosynthetically labelled in vitro using {35S}methionine and proteins were immunoprecipitated with rabbit anti-ookinete serum or monoclonal antibodies . Early zygotes (approx . 2 h post-gametogenesis and fertilization) synthesized and expressed on their surface a protein of Mr 26 000 as observed under reducing conditions on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) (31 000 under non-reducing conditions) and continued to do so for 8-10 h; thereafter synthesis of the Mr 26 000 protein declined and little or none was synthesized in the mature ookinetes (greater than 20 h post-gametogenesis) . Between 3-5 h post-gametogenesis, zygotes also began to synthesize a protein of Mr 28 000 (34 000 under non-reducing conditions) . Synthesis and expression of this surface protein continued throughout development; and the Mr 28 000 protein was the predominant surface protein synthesized by the mature ookinete . Mr 26 000 and Mr 28 000 proteins have been designated earlier as PgO-1 and PgO-2 respectively (Carter and Kaushal, Mol . Biochem . Parasitol . (1984) 13, 235-241) . Neither protein was synthesized in the gametocytes prior to gametogenesis . Both proteins could be labelled with {3H}glucosamine or {3H}mannose . When zygotes were incubated with {3H}palmitic acid both PgO-1 and PgO-2 bound fatty acids in covalent linkage . The two proteins do not otherwise appear to be structurally related . They were differentially immunoprecipitated by different monoclonal antibodies and gave rise to distinct patterns of peptides following digestion with proteases such as Staphylococcus aureus V-8, trypsin and chymotrypsin. J Antimicrob Chemother, 1985 Feb, 15(2), 219 - 32 Efficacy of lincosaminide antibiotics in the treatment of experimental staphylococcal mastitis in lactating mice; Yancey RJ Jr et al.; Staphylococcus aureus is a frequent cause of bovine mastitis worldwide . A model that may predict the efficacy of antimicrobial agents in the treatment of bovine mastitis induced by Staph . aureus was developed in lactating mice . Infection was established by the inoculation of lactating CF1 mice with Staph . aureus into the mammary gland via the teat duct . At the dose of bacteria used, 85-90% of the inoculated, untreated animals developed a nonlethal, acute mastitis within 48 h . Antibiotic treatment was administered subcutaneously or by the intramammary route . Lincosaminide antibiotics including lincomycin, clindamycin, and pirlimycin were evaluated in this system . Other compounds which have been used in therapy of bovine mastitis including novobiocin, penicillin G, ampicillin, cloxacillin and rifamycin-SV were used as reference antibiotics . Pirlimycin was the most effective of the antibiotics tested in this standardized system . Depending upon the route of administration, this novel lincosaminide was 15 to 95-fold more effective than clindamycin, three- to six-fold better than lincomycin, two- to ten-fold more effective than novobiocin, 13- to 17-times more effective than cloxacillin and 8- to 22-times better than rifamycin-SV on a weight-dose comparison . Penicillin G and ampicillin were the least effective drugs tested against mastitis induced by the beta-lactamase producing strain of Staph . aureus used in these assays . Pharmacokinetic experiments suggested that the greater effectiveness of pirlimycin compared to clindamycin and lincomycin was due to increased affinity for and prolonged retention in the mammary gland. J Virol, 1985 Feb, 53(2), 679 - 83 Virions and intracellular nucleocapsids produced during mixed heterotypic influenza infection of MDCK cells; Sklyanskaya EI et al.; Phenotypically mixed virus yields, obtained by coinfection of MDCK cells with influenza A/WSN/33 and B/Lee/40 viruses, contained both A/WSN/33 and B/Lee/40 NP proteins, as revealed by polyacrylamide gel electrophoresis of the purified 14C-amino acids-labeled virus . Virions were lysed with 0.6 M KCl-Triton X-100 buffer, and nucleocapsids were immunoprecipitated with antibodies against NP protein of influenza A virus . Polyacrylamide gel electrophoresis of the immunoprecipitate revealed NP protein of A/WSN/33 but not of B/Lee/40 virus . However, in similar experiments with the lysates of doubly infected cells, the band of B/Lee/40 NP protein was revealed in the polyacrylamide gel electrophoresis patterns of the immunoprecipitates . In an attempt to analyze the RNA content of the immune complexes, we absorbed the lysates of doubly infected {3H}uridine-labeled cells with protein A-containing Staphylococcus aureus covered with antibodies against the NP protein of influenza A virus . RNA extracted from the immune complexes contained genomic RNA segments of both A/WSN/33 and B/Lee/40 viruses . In control samples containing an artificial mixture of cell lysates separately infected with each virus, the analysis revealed homologous components (i.e., A/WSN/33 NP protein or RNA segments) in the immune complexes . The results suggest the presence of phenotypically mixed nucleocapsids in the cells doubly infected with influenza A and B viruses; in the course of the virion formation, the nucleocapsids lacking the heterologous NP protein are selected. Aust J Exp Biol Med Sci, 1985 Feb, 63 ( Pt 1), 19 - 32 Regulation of immunity and tolerance to type III pneumococcal polysaccharide (SIII) by functionally distinct IgM anti-SIII antibodies; Kearney R et al.; When BALB/c mice and athymic (nude) mice are injected intraperitoneally (i.p.) with pneumococcal type III polysaccharide (SIII), their antibodies as measured by passive haemagglutination (HA) are inhibited more easily by high doses of SIII than antibody measured by passive haemolysis (HL) . The HA activity, due mainly to a highly avid non-complement-fixing (NCF) type of IgM, was further distinguished from the HL activity (CF-IgM, or CF-IgM plus CF hybrid IgM/A anti-SIII antibodies) by the failure of the NCF-IgM anti-SIII to bind to protein-A of Staphylococcus aureus (Sa) . High-dose tolerance in the HL anti-SIII antibody response of BALB/c and athymic mice was induced only in the absence of circulating NCF-IgM anti-SIII antibodies . The presence of NCF-IgM anti-SIII antibodies formed to multiple daily increasing amounts of SIII, commencing with 0.01 micrograms SIII, decreased the magnitude of the HL anti-SIII response to subsequent daily increments of SIII antigen injected into BALB/c and athymic (nude) mice . Thus, the effect on the HL anti-SIII response was independent of T-cells . The concomitant administration of NCF-IgM anti-SIII rendered SIII less tolerogenic in primed mice . In contrast to the HL activity, the NCF-IgM anti-SIII antibodies were induced to low doses of SIII, conferred protection against viable pneumococci, but did not precipitate the soluble antigen in agar . It is proposed that immune paralysis (as defined by the failure of SIII-injected mice to resist pneumococcal challenge) is not necessarily a condition of total unresponsiveness but is due to an absence of protective NCF-IgM anti-SIII antibodies . Thus, immune paralysis can co-exist with either the presence or absence of non-protective CF-IgM or CF-IgM/A anti-SIII antibodies. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1985 Feb, 259(1), 71 - 7 Purification of oligomeric staphylococcal alpha-toxin by affinity chromatography on digitonin-sepharose; Schaeg W et al.; An effective concentration of alpha-toxin from Staphylococcus aureus Wood 46, directly from the culture supernatant, could be achieved by adsorption on digitonin-sepharose and elution with 3 mol/l sodium thiocyanate (NaSCN) . The toxin was further purified by gelchromatography . The purified product yielded 1 single protein band upon SDS-polyacrylamide electrophoresis . It was nonhemolytic, but reacted with anti-alpha-toxin under complement fixation . Dialysis against 0.14 mol/l NaCl with hydrophobic amino acids partially reactivated the alpha-hemolytic activity of the toxin . Ultracentrifugal analysis yielded sedimentation coefficients for the purified toxin of approximately 3,7 S when dissolved in 3 mol/l NaSCN and of about 12 S after dialysis against 0.14 mol/l NaCl (Table 1) . The spontaneous oligomerization of the alpha-toxin during dialysis against 0.14 mol/l NaCl possibly resulted from a change in configuration induced by its adsorption to digitonin-sepharose. Immunobiology, 1985 Feb, 169(1), 11 - 20 Lipopolysaccharide significantly enhances erythrophagocytosis but marginally stimulates the phagocytosis of Staphylococcus aureus in mouse peritoneal macrophages; Lokesh BR et al.; The influence of lipopolysaccharide (LPS) on phagocytic and bactericidal functions of normal mouse peritoneal macrophages was investigated . Preincubation of macrophages with LPS enhanced their capacity for phagocytosis of antibody coated sheep red blood cells 5-fold, but phagocytosis of antibody coated Staphylococcus aureus was enhanced only 1.2-ld . Phagocytosis and intracellular killing of unopsonised or normal rabbit serum opsonised S . aureus was not affected by the LPS treatment of macrophages. Contraception, 1985 Feb, 31(2), 185 - 94 In vitro amplification of toxic shock syndrome toxin-1 by intravaginal devices; Tierno PM Jr et al.; Super-absorbent tampons and an exotoxin of Staphylococcus aureus have been associated with the recent emergence of toxic shock syndrome (TSS) . In the majority of cases, when a TSS strain of S . aureus was cultivated in the presence of various tampons and a contraceptive sponge, increased amounts of toxic shock syndrome toxin-1 (TSST-1) were observed to be produced into the blood medium by the bacterium . The amplification of toxin by these products adds support to the epidemiologic data in establishing the causal link between tampons and TSS. Burns Incl Therm Inj, 1985 Feb, 11(3), 202 - 6 Survival of an extensively burned infant following purulent pericarditis; Nakamura K et al.; This is a report of the treatment and survival of an extensively burned infant following purulent pericarditis with massive pericardial effusion due to Staphylococcus aureus . A 2-year-old boy fell into a bathtub and suffered scalds covering at least 70 per cent of the body surface area . Pericarditis with massive pericardial effusion was diagnosed on post-burn day 36 . As conservative treatment was ineffective pericardiotomy and pericardial drainage were carried out . Whole body oedema disappeared promptly and entirely and the patient was discharged from hospital with healed burns and free of cardiac symptoms. Antimicrob Agents Chemother, 1985 Feb, 27(2), 181 - 3 Once-daily ceftriaxone therapy for serious bacterial infections in children; Congeni BL et al.; Ceftriaxone administered as a single daily dose of 50 mg/kg was evaluated in the treatment of 35 children with a variety of nonmeningitic bacterial infections . In two of the patients, the drug was discontinued before the response to the drug could be evaluated . All of the remaining patients had a satisfactory response . In 22 of the patients, plasma was available for the determination of ceftriaxone levels 1 h after a dose and immediately before the next dose . All but one of these patients had trough ceftriaxone levels which exceeded the MIC of the infecting organism, although marginally so for Staphylococcus aureus . Ceftriaxone appears to be safe and effective in the treatment of a variety of bacterial pathogens in children when administered at a single daily dose of 50 mg/kg . This drug may be especially useful in those patients in whom outpatient antibiotic therapy is contemplated or in whom maintenance of intravenous access is difficult. J Gen Microbiol, 1985 Feb, 131 ( Pt 2), 405 - 8 A comparison of the patterns of extracellular proteins produced by the high alpha-toxin-secreting organism Staphylococcus aureus (Wood 46) during aerobic and anaerobic growth; Coleman G; Staphylococcus aureus (Wood 46) was grown aerobically and anaerobically in supplemented 3% (w/v) Tryptone Soya Broth medium for 24 h at 37 degrees C . Although the bacterial density achieved was 9 times higher in the aerobic culture, the exoprotein produced per unit of bacterial dry weight was only 1.4 times higher than in the anaerobic culture . However, the SDS-PAGE patterns of extracellular proteins were quite different: the aerobic products occurred almost exclusively in the mol . wt range 15-30000 compared with 30-60000 for those produced anaerobically . The only major component common to both preparations was alpha-toxin which accounted for 2.4 times more of the total exoprotein under aerobic than under anaerobic conditions. South Med J, 1985 Feb, 78(2), 157 - 8 Hand infections in the elderly; Stromberg BV; Diagnosis and management of hand infections in the elderly can be challenging . The general principles of rest, elevation, compresses, and drainage when appropriate apply . Antibiotics are important to therapy . Review of data from elderly patients and comparison with a younger population having identical infections show a number of important differences . Temperature, pulse, and white blood cell and differential counts were not elevated significantly enough to be useful . Culture data show fewer pure Staphylococcus aureus infections (20%) and fewer pure gram-positive infections (20%) than the 34% and 56% respectively in a younger population . On the other hand, there were significantly more mixed gram-positive and gram-negative infections (60%) . Significantly, the average number of organisms per infection is increased (2.4 vs 1.9 per infection) . Antibiotic susceptibility is significantly worse . The cephalosporins and the penicillinase-resistant antibiotics remain good choices. Scand J Immunol, 1985 Feb, 21(2), 189 - 93 Adherence of lysostaphin to and penetration into human monocytes; van den Broek PJ et al.; The effect of lysostaphin on Staphylococcus aureus phagocytosed by monocytes was investigated . The results showed that lysostaphin adheres to monocytes by a temperature-independent mechanism, is not adequately removed from monocytes by washing, and penetrates by means of a temperature-dependent mechanism . In in vitro assays of monocyte function, phagocytosed S . aureus can be killed by lysostaphin after penetration of the cells during incubation or by adhering lysostaphin when the monocytes are disrupted. J Cell Sci, 1985 Feb, 73, 279 - 97 Biochemical evidence for the presence of an actin protein in Tetrahymena pyriformis; Mitchell EJ et al.; A protein from an ATP extract prepared from an acetone powder of Tetrahymena pyriformis GL was identified as actin . The protein migrated slightly behind muscle actin on sodium dodecyl sulphate (SDS)/10% polyacrylamide gels (SDS/PAGE) with an apparent molecular weight of 47 500 (47.5 X 10(3) Mr) . Partial proteolysis of this band with Staphylococcus aureus V-8 protease followed by electrophoresis revealed a pattern of peptides in which at least four peptides were similar to those observed after digestion of rabbit skeletal muscle actin . The 47.5 X 10(3) Mr protein appeared particularly susceptible to endogenous proteolytic cleavage, which was inhibited by leupeptin . An ATP extract prepared with leupeptin was applied to a DNase I-affinity column and a distinct peak was eluted with 3 M-guanidine . HCl; the DNase I-binding protein appeared as a distinct band on SDS/PAGE with an apparent molecular weight of 47.5 X 10(3) Mr . In the absence of leupeptin, the DNase I-binding protein appeared as a broad 34 X 10(3) Mr band on gels . Both the ATP extract and the DNase I-binding protein showed reactivity with commercially available antiserum raised against native chicken skeletal muscle actin as determined by an enzyme-linked immunosorbance assay (ELISA) . Immuno-blotting studies and affinity purification of this antiserum showed that the recognition was not specific to the 47.5 X 10(3) Mr protein . However, using affinity-purified anti-actin antibodies raised against denatured actin from chick smooth muscle, recognition of the 47.5 X 10(3) Mr protein and a 34 X 10(3) Mr protein was shown . In negatively stained preparations from an ATP extract after two cycles of polymerization and depolymerization there were filaments, 8-12 nm diameter, which did not decorate with subfragment S-1 of myosin, but which resembled intermediate filaments . Analysis of these filaments on SDS/PAGE indicated an intensely stained 54 X 10(3) Mr band . It is suggested that, in vitro, Tetrahymena intermediate filaments assemble under conditions expected to assemble actin filaments . Thus, in Tetrahymena there is a protein that resembles actin in its extractability, molecular weight, peptide pattern after partial proteolysis, DNase I-binding capacity and reactivity with anti-actin antibodies . However, this protein did not assemble into actin filaments in crude extracts. Exp Eye Res, 1985 Feb, 40(2), 327 - 34 In vitro reassociation of EDTA-extractable proteins with calf lens fiber membranes; van den Eijnden-van Raaij AJ et al.; Nature and site of membrane binding of the EDTA-extractable proteins (EEP) from calf lens fiber membranes have been studied . Reassociation of EEP to EEP-free lens fiber membranes only occurs by means of calcium, not by magnesium ions . This EEP-membrane binding is not hindered by the cytoskeleton . The proportional distribution of the EEP protein components is not altered by reattachment of EEP to the membrane . Calcium appears to be a limiting factor in the reassociation of EEP with the membrane . The total amount of reassociated EEP increases with increasing calcium concentration and may largely exceed the quantity of naturally occurring EEP in lens fiber membranes . In addition, the latter EEP-containing membranes are able to bind an additional amount of EEP in the presence of calcium . These results indicate that most of the EEP-binding sites in lens fiber membranes are not occupied by EEP . Trypsin- or Staphylococcus aureus V8 protease-treated fiber membranes retain the capacity to bind EEP in the presence of calcium . This result indicates that the small polypeptide fragment of the main intrinsic protein (MIP), which is accessible to proteolytic attack, very likely is not the membrane attachment point for EEP . It is suggested that phospholipids rather than membrane proteins are involved in the calcium-dependent binding of EEP to calf lens fiber membranes. Exp Cell Res, 1985 Feb, 156(2), 429 - 38 Significant non-S-phase DNA synthesis visualized by flow cytometry in activated and in malignant human lymphoid cells; Neckers LM et al.; The development of a monoclonal antibody to the deoxynucleoside bromodeoxyuridine (BrdU), combined with two parameter flow cytometry, has allowed us to examine large numbers of cells for non-S-phase DNA synthesis . Three human lymphoid cell populations were studied to determine the level of deoxynucleoside (dN) incorporation as a function of DNA content . In each population, non-S-phase DNA synthesis was observed . In a rapidly growing human T-lymphoblastoid cell line (CCRF-CEM), 53% of dN incorporation occurred in G0/G1 plus G2 + M . In chronic lymphocytic leukemia (CLL) cells stimulated with tetradecanoylphorbol acetate (TPA), 45% of the observed burst in thymidine incorporation was found to be localized to G0/G1 cells . Non-S-phase incorporation was not, however, limited to neoplastic cells . Normal human peripheral blood B cells treated with the Cowan strain of Staphylococcus aureus (CSA) undergo a transient burst in thymidine incorporation, but do not go on to divide in the absence of other stimuli . Flow-cytometric analysis showed that 80% of this CSA-stimulated dN incorporation was into G0/G1 cells . These data are consistent with a more dynamic state of DNA synthesis than usually envisioned . Furthermore, the data show that although thymidine incorporation levels are related to incorporation of dN into DNA, they can be unrelated to cell proliferation. Anal Biochem, 1985 Feb 1, 144(2), 522 - 6 Peptide mapping of basic proteins by proteolysis in acetic acid/urea-minislab polyacrylamide gels; Davie JR; A method to obtain peptide maps of basic proteins on acetic acid/urea (AU) -polyacrylamide minislab gels is presented . Basic proteins such as the histones are digested with Staphylococcus aureus V8 protease in the stacking gel (pH 4) of an AU-polyacrylamide minislab gel . As the peptides are resolved in the AU minislab gel on the basis of charge and size, it is possible to separate peptides containing modified amino acids from the unmodified, parent peptide . The peptide(s) containing the modified residue may be identified following electrophoresis on a second-dimension sodium dodecyl sulfate-polyacrylamide minislab gel . This procedure will be useful for comparing histone variants and for the study of histone modifications. Am Fam Physician, 1985 Feb, 31(2), 131 - 7 Pneumonia in the elderly: a nursing home perspective; Roth RM et al.; The development of pneumonia is a life-threatening event in a nursing home resident . Staphylococcus aureus and Klebsiella pneumoniae are major identifiable bacterial pathogens . Some elderly patients with pneumonia can be effectively treated in the nursing home . The clinical impression of pneumonia merits radiologic confirmation . Cefuroxime, trimethoprim-sulfamethoxazole and cefaclor offer theoretic advantages in the treatment of bacterial pneumonia in elderly nursing home residents. J Immunol, 1985 Feb, 134(2), 1153 - 9 Effect of antibodies directed against complement receptors on phagocytosis by polymorphonuclear leukocytes: use of iodination as a convenient measure of phagocytosis; Klebanoff SJ et al.; Two monoclonal antibodies (Mab), designated 60.3 and 60.1, markedly inhibited the phagocytosis of serum-opsonized zymosan by human polymorphonuclear leukocytes (PMN) as measured by the iodination reaction and by microscopic visualization . These antibodies also inhibited rosette formation with EC3bi without decreasing EC3b rosetting, suggesting that Mab 60.3 and 60.1 inhibit the phagocytosis of opsonized zymosan through reaction with the C3bi receptor (CR3) on the leukocyte surface . In support of this concept is the finding that the PMN of two patients with recurrent infections do not ingest opsonized zymosan, lack C3bi receptor function, and react weakly or not at all with Mab 60.3 and 60.1 . At concentrations which completely inhibited ingestion of opsonized zymosan, both Mab partially inhibited iodination with Staphylococcus aureus 502A as the particle, and did not affect iodination when Staphylococcus epidermidis was used . This presumably reflects a variable need among the opsonized particles for CR3 for ingestion . Mab 60.3 also inhibited the phagocytosis of certain unopsonized particles as measured by iodination, indicating that the antigens recognized by the Mab do not influence phagocytosis solely by functioning as a C3bi receptor . Mab 60.3 increased the phagocytosis of unopsonized, heat-killed S . aureus by reaction with the PMN via its antibody-combining site, and with the staphylococcal protein A via its Fc region (reverse opsonization) . This process required protein A-containing organisms (S . aureus 502A or Cowan 1 but not S . aureus Wood 46 or S . epidermidis), was inhibited by purified protein A, and was not seen either when the F(ab')2 or Fab fragments of the antibody, or when PMN which lack or have low levels of the antigen were employed . Thus, these studies, using iodination as a convenient method for the measurement of phagocytosis, demonstrated two effects of antibodies directed against PMN cell surface components: inhibition of phagocytosis by reaction with the C3bi receptor, and stimulation of phagocytosis by reverse opsonization. Scand J Immunol, 1985 Feb, 21(2), 141 - 50 Abnormal production of and response to B-cell growth factor and B-cell differentiation factor in patients with systemic lupus erythematosus; Hirose T et al.; We examined the production of and the response to B-cell growth factor (BCGF) and B-cell differentiation factor (BCDF) in 21 patients with systemic lupus erythematosus (SLE) and 23 normal subjects . T cells, 2.5 X 10(6)/ml, were cultured for 24 or 72 h with 1% phytohaemagglutinin (PHA) . After absorption of PHA by chicken erythrocytes (CRBC), they were used for BCGF and BCDF . In inactive SLE, BCGF activity was significantly lower than that in normal subjects . Active SLE contained two separate groups, one showing normal BCGF activity and the other showing lower activity than normal . In contrast, BCDF activity from initial culture in active SLE was elevated . The B-cell response both to BCGF and BCDF was elevated in active SLE without Staphylococcus aureus Cowan I antigen (SAC) preactivation . However, the B-cell response to SAC was markedly disturbed . Thus SLE B cells were shifted to the mature state in vivo . We also demonstrated pivotal abnormalities of monocytes in SLE B-cell growth and differentiation . These results may contribute to the understanding of the abnormalities of T-B interactions and the overproduction of antibody in SLE. J Clin Invest, 1985 Feb, 75(2), 754 - 61 Effects of in vitro corticosteroids on B cell activation, proliferation, and differentiation; Cupps TR et al.; The present study demonstrates the graded effect of in vitro corticosteroids (CSs) on the different phases of B cell activation, proliferation, and differentiation . Early events such as activation and proliferation of high-dose anti-mu or Staphylococcus aureus-stimulated B cells are profoundly suppressed by the presence of in vitro CSs . The suppressed proliferative response may be mediated by a direct effect on B cells and/or modulation of accessory cell function . Later events in the B cell cycle such as the proliferative response to B cell growth factor after either in vivo or in vitro activation are less sensitive to the suppressive effects of in vitro CSs . The final events in the B cell cycle; namely, the differentiation to the immunoglobulin-producing state, is not suppressed by in vitro CSs . Indeed, depending on the systems employed, there is either no effect or enhancement of immunoglobulin secretion by the presence of in vitro CSs . The graded effect of in vitro CSs on the discrete phases of the B cell activation, proliferation, and differentiation cycle provide new insights into the complex nature of CS-induced modulation of human B cell responses. Eur J Immunol, 1985 Feb, 15(2), 193 - 6 B cell growth factor activity of immunoaffinity-purified and recombinant human interleukin 2; Mingari MC et al.; We investigated the effect of recombinant and affinity-purified human interleukin 2 (IL2) on human B cell proliferation . Five X 10(4) nonadherent spleen cells that had been depleted twice of T cells were activated by 3-day culture with formaldehyde-killed Staphylococcus aureus Cowan strain I (SAC) prior to addition of tested growth factors . Cultures were harvested 72 h later . It was found that both IL2 preparations led to optimal cell proliferation compared with a control supernatant obtained by 36 h phytohemagglutinin stimulation of spleen mononuclear cells . Moreover, the effect of such spleen supernatant on B cell proliferation correlated with the IL2 activity since its B cell growth factor activity (BCGF) was not greater than that of purified IL2 and no residual BCGF activity could be detected after absorption of all IL2 activity by the IL2-dependent cytotoxic T lymphocyte line cells . T cells, enumerated as E-rosetting cells as well as T3+ cells, represented 0.2 to 2% of the cells recovered at termination of the cultures (day 6) and there were less than 1% E-rosetting cells in freshly purified or SAC-activated (day 3) B cell populations . Therefore, we conclude that IL2 is a growth factor not only for activated T cells but also for activated human B cells. Infect Immun, 1985 Feb, 47(2), 514 - 21 Penicillinase plasmid-linked genetic determinants for enterotoxins B and C1 production in Staphylococcus aureus; Altboum Z et al.; The genes encoding for beta-lactamase (bla+) and resistance to metallic ions (cadmium, mercury, lead, arsenate, and arsenite) were located in a 56.2-kilobase plasmid, pZA10, isolated from a clinical strain, Staphylococcus aureus 6344 . This strain produced enterotoxin B and enterotoxin C1 . Elimination of pZA10 by either sodium dodecyl sulfate or heat treatment (43 degrees C) resulted in the loss of the capability of the bacteria to produce both enterotoxin B and enterotoxin C1 . A physical map of pZA10 was constructed with BamHI, SalI and BglII restriction endonucleases . Penicillin-resistant, enterotoxin B- and C1-producing cotransformants were isolated by transformation with pZA10 DNA with either S . aureus RN450 or cured S . aureus 6344 as recipients . The transferred plasmids exhibited genetic instability shown by changes in restriction pattern and molecular size, loss of plasmid DNA, and addition of chromosomal DNA . Enterotoxin B production was related to a 18.1-kilobase pZA10 fragment carried by such a rearranged plasmid . Chromosomal cointegration of bla+ with genetic determinants for metallic ion resistance and enterotoxin B and C1 production were detected in heat-treated S . aureus 6344 . Transformation employing chromosomal DNA containing the integrated plasmid resulted in excision and reestablishment of pZA10-related plasmids in the transformants . pZA10-linked resistance to cadmium, which was lost upon the integration of pZA10 into the host chromosome, reappeared in transformants carrying the excised plasmid. EMBO J, 1985 Feb, 4(2), 561 - 8 The use of synthetic oligonucleotides with universal templates for rapid DNA sequencing: results with staphylococcal replicon pC221; Brenner DG et al.; A rapid sequencing strategy has been devised and applied to determine the complete nucleotide sequence (4555 bp) of Staphylococcus aureus plasmid pC221 . The entire replicon was cloned into phage M13mp8 in both orientations to provide 'universal templates' for primed DNA synthesis from internally-sited oligonucleotide primers . The latter were synthesized by a modification of a recently described paper disc method which employs phosphotriester chemistry . Less than 4 weeks was required for the synthesis of the required primers and for the sequencing experiments . Plasmid pC221 bears a substrate-inducible chloramphenicol acetyltransferase (CAT) gene that shares much homology with its counterparts in pC194 (S . aureus) and the chromosomal cat-86 gene of Bacillus pumilus, both in coding regions and upstream sequences believed to be involved in the induction phenomenon . A second plasmid-specified protein, REP D, has an 81% identity in the REP C polypeptide that has been shown to be essential for the replication of staphylococcal plasmid pT181 . The 5' flanking region of rep D shows striking similarities with its counterpart in rep C that determines copy number and incompatibility . The nucleotide sequence reveals two additional and overlapping open reading frames that may specify proteins that play roles in plasmid relaxation and transfer. Jpn J Antibiot, 1985 Feb, 38(2), 199 - 202 {Antibiotic susceptibility of the clinically isolated staphylococcal strains resistant to cephalosporin derivatives}; Arai T; Incidence of cephalosporin-resistant staphylococcal infections is increasing recently . We tried to find out the possible first choice antibiotic for these infections . We estimated minimal inhibitory concentrations (MIC) of each one of the broad spectrum antibiotics with different mode of actions as well as representative drugs of penicillins and cephalosporins against strains of Staphylococcus epidermidis and Staphylococcus aureus . Trend of antibiotic susceptibility of S . epidermidis was found to be as same as that of S . aureus . MIC of minocycline was found to be the lowest in the drugs tested, and there found no resistant strains . MIC of erythromycin was next low but more than a half strains were found to be resistant to this drug . Some of the strains were thought to be treated with amikacin or sulfonamides by the MIC against them, but there also found many resistant strains . Therefore, use of these drugs should be decided after sensitivity test of the causative bacteria . Most of the strains were found not to be treated with beta-lactam antibiotics . In conclusion, minocycline could be the only one first choice drug for staphylococcal infections before antibiotic susceptibility test of the causative strains in the present moment. J Antimicrob Chemother, 1985 Feb, 15(2), 201 - 7 Differences in ability of cell-wall antibiotics to suppress emergence of rifampicin resistance in Staphylococcus aureus; Eng RH et al.; Rifampicin resistance developed easily in methicillin-susceptible and methicillin-resistant strains of Staphylococcus aureus during an overnight incubation in broth containing 0.1 mg/l of rifampicin . Incubation of methicillin-susceptible Staph . aureus and 0.1 mg/l of rifampicin with 1 mg/l of nafcillin reduced the emergence of rifampicin resistance with only 5 of 50 strains (10%) becoming rifampicin-resistant . However, incubation of the methicillin-susceptible or methicillin-resistant strains with 0.1 mg/l of rifampicin and 1 mg/l of vancomycin did not prevent the development of rifampicin resistance . Rifampicin resistance developed in 25 of 50 (50%) of methicillin-susceptible and 32 of 50 (64%) methicillin-resistant Staph . aureus strains tested . These data would suggest that differences exist in the abilities of nafcillin and vancomycin to suppress the development of rifampicin resistance in Staph . aureus (P less than 0.01) . Caution should be exercised when the combination of vancomycin and rifampicin is used for infections caused by Staph . aureus and Staph . aureus isolates recovered during therapy should be monitored for the development of rifampicin resistance. J Clin Microbiol, 1985 Feb, 21(2), 205 - 10 Effect of the source of Mueller-Hinton agar and resistance frequency on the detection of methicillin-resistant Staphylococcus aureus; Hindler JA et al.; Inconsistencies in the results of disk diffusion tests of oxacillin against Staphylococcus aureus that occurred when using commercially prepared Mueller-Hinton agar from different sources led us to evaluate the ability of media from different sources to detect resistance to oxacillin, methicillin, and nafcillin in S . aureus . Mueller-Hinton agar from five manufacturers was prepared in our laboratory and used for standard disk diffusion and agar dilution tests . Ten oxacillin-resistant S . aureus isolates, of which three were definitive-resistant and seven were occult-resistant, were examined . All definitive-resistant strains were resistant to all three antimicrobial agents on four out of five agars . The occult-resistant strains were consistently detected as resistant on only one of the agars . With only slight differences, oxacillin, methicillin, and nafcillin resistance was more readily detected by disk diffusion and agar dilution when initially incubated at 30 degrees C, and extended incubation improved the detection . The frequency of resistance within a population of occult-resistant cells was low compared with the frequency within a population of definitively resistant cells . The heterogeneity of colony morphology and apparent growth rates within a population of occult-resistant cells contributed to the problem of detecting some resistant isolates . Definitive-resistant isolates were characterized by a very high and stable frequency of resistance . Occult-resistant strains were characterized by a lower frequency of resistance, although the true frequency of resistance may be difficult to ascertain because of heterogeneity in growth rates. J Antimicrob Chemother, 1985 Feb, 15(2), 173 - 80 In-vitro activity of methicillin against clinical isolates of Staphylococcus aureus; Frimodt-Moller N et al.; Methicillin activity against 149 penicillin-resistant, methicillin-susceptible Staphylococcus aureus strains from bacteraemia cases with endocarditis (n = 89) or without endocarditis (n = 60), from the years 1976-1981, was studied with broth dilution and agar dilution . While no differences in methicillin susceptibility were found in relation to the origin of the strains, Staph . aureus of the phage type complex 94,96 showed significantly higher MIC and IC50 by agar dilution than strains of other phage groups/complexes . This difference probably has no clinical importance but is of epidemiological interest . Broth dilution MIC was generally one dilution higher than agar dilution MIC, possibly explained by methodological factors . The MBC/MIC ratios never exceeded two in any of the strains, indicating a lack of tolerance in these clinically important isolates. J Am Acad Dermatol, 1985 Feb, 12(2 Pt 1), 319 - 24 Treatment of gram-negative folliculitis with isotretinoin: positive clinical and microbiologic response; James WD et al.; Thirty-two patients with gram-negative folliculitis were treated with 0.47 to 1.0 mg/kg/day of isotretinoin . Serial microbiologic evaluations demonstrated rapid clearing of the face and nasal mucosa of gram-negative rods . The clinical response was rapid, complete, and induced prolonged remissions . Twenty-six of thirty-two patients developed Staphylococcus aureus nasal carriage by the end of the 20-week treatment course . Isotretinoin has decided advantages over previously reported therapies for gram-negative folliculitis. Infect Immun, 1985 Feb, 47(2), 502 - 7 Relationship between extracellular stimulation of intracellular killing and oxygen-dependent microbicidal systems of monocytes; Leijh PC et al.; Human monocytes require serum components immunoglobulin G, C3/C3b, and B/Bb to exert optimal microbicidal action against ingested microorganisms . The present study was performed to find out whether these factors act by enhancing oxygen-dependent antimicrobial mechanisms . Serum enhanced oxygen consumption and superoxide production by monocytes before phagocytosis, but did not further increase these processes in monocytes that had recently ingested bacteria . Furthermore, serum did not boost iodination during intracellular killing by monocytes . Phorbol myristate acetate, N-formyl-methyonyl-leucyl-phenylalanine, concanavalin A, and concanavalin A-Sephadex all stimulated the conversion of O2 to H2O2 by monocytes, but only concanavalin A augmented intracellular killing . Reactive oxygen intermediates generated by cell-free enzymes (xanthine oxidase or glucose oxidase) in concentrations comparable to those accumulating extracellularly during incubation of monocytes containing bacteria with phorbol myristate acetate did not promote intracellular killing . The presence of catalase during phagocytosis inhibited killing, but had no effect on killing in the postphagocytic state . Monocytes deprived of glucose for 24 h showed markedly impaired O2 consumption, O2- generation, and bacterial killing; all of these effects were rapidly reversed by restoration of glucose . It is concluded that both an intact respiratory burst and extracellular serum factors are necessary for optimal killing of intracellular Staphylococcus aureus by human monocytes . Serum does not appear to act by enhancing the respiratory burst, but rather to have a separate, synergistic role, the biochemical basis of which is unknown. Proc Natl Acad Sci U S A, 1985 Feb, 82(3), 638 - 42 Plasmid pT181 replication is regulated by two countertranscripts; Kumar CC et al.; A transcription map of the replication control region of the Staphylococcus aureus plasmid pT181 has been constructed . Two major leftward transcripts, RNA III and RNA IV, start at positions 339 and 413, respectively . These two RNAs can serve as mRNAs for a plasmid-specific replication protein RepC . Two short rightward transcripts, RNA I and RNA II, approximately 85 and 150 nucleotides long, respectively, start at position 246 . These rightward transcripts (referred to as countertranscripts) do not appear to be translated but act directly as negative regulators of plasmid replication, probably by interfering with translation of the RepC mRNAs . There is no significant base sequence homology among the countertranscripts of pT181, ColE1, and R1/NR1/R6-5, suggesting that the structural parallelism has risen by convergent molecular evolution. Biochemistry, 1985 Jan 29, 24(3), 776 - 81 Inhibition of monoclonal antibody binding and proteolysis by light-induced phosphorylation of rhodopsin; Molday RS et al.; Light-induced phosphorylation of rhodopsin in bovine rod outer segment disk membranes inhibits the binding of three carboxyl-terminal-specific anti-rhodopsin antibodies and the cleavage of the carboxyl-terminal region of rhodopsin by trypsin and Staphylococcus aureus V-8 protease . Two monoclonal antibodies, rho 3A6 and rho 1C5, which previously have been shown to preferentially bind to the 8'-12' and the 9'-18' carboxyl-terminal segments of rhodopsin, respectively, are both highly sensitive to phosphorylation . When an average of one phosphate is incorporated per rhodopsin, the binding reactivity of rhodopsin for these antibodies decreases to 30% that of nonphosphorylated rhodopsin as measured in radioimmune competition assays . Reactivity of the rho 1D4 antibody whose primary binding site is localized in the 1'-8' C-terminal segment of rhodopsin is unaffected at this level of phosphorylation but decreases to 30% when three phosphates on average are incorporated per rhodopsin . Direct binding studies using 125I-labeled antibodies indicate that phosphorylation of rhodopsin decreases the maximum extent of rho 3A6 and rho 1C5 binding to rhodopsin . For rho 1D4, the maximum extent of binding is unaffected by phosphorylation, but the dissociation constant is increased by 10-fold . Phosphorylation of rhodopsin also inhibits cleavage of the 1'-9' and 1'-7' carboxyl-terminal peptides by trypsin and S . aureus V-8 protease, respectively . When an average of one phosphate per rhodopsin is incorporated, cleavage decreases to 40% that of nonphosphorylated rhodopsin as measured by high-performance liquid chromatography . Phosphorylation of rhodopsin had no effect on S . aureus cleavage of rhodopsin into the F1 (Mr 25 000) and F2 (Mr 12 000) fragments.(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1985 Jan 25, 260(2), 834 - 40 The amino acid sequence of Acanthamoeba profilin; Ampe C et al.; The complete amino acid sequence of Acanthamoeba profilin was determined by aligning tryptic, chymotryptic, thermolysin, and Staphylococcus aureus V8 protease peptides together with the partial NH2-terminal sequences of the tryptophan-cleavage products . Acanthamoeba profilin contains 125 amino acid residues, is NH2-terminally blocked, and has trimethyllysine at position 103 . At five positions in the sequence two amino acids were identified indicating that the amoebae express at least two slightly different profilins . Charged residues are unevenly distributed, the NH2-terminal half being very hydrophobic and the COOH-terminal half being especially rich in basic residues . Comparison of the Acanthamoeba profilin sequence with that of calf spleen profilin (Nystrom, L . E., Lindberg, U., Kendrick-Jones, J., and Jakes, R . (1979) FEBS Lett . 101, 161-165) reveals homology in the NH2-terminal region . We suggest, therefore, that this region participates in the actin-binding activity. Med J Aust, 1985 Jan 21, 142(2), 138 - 9 Hospital outbreak of multiresistant Staphylococcus aureus in Victoria, 1979-1984; Bennett NM; The outbreak of nosocomial infection caused by multiresistant Staphylococcus aureus (MRSA) strains in Victoria has been monitored by a continuing programme of collecting monthly statistics from all hospitals on new patients who have been either colonized or infected by these strains . Data collected during the period from November 1980 to September 1984 have been plotted on graphs . Whereas the prevalence of MRSA strains has remained virtually unchanged, the number of infections attributed to them appears to have gradually decreased since November 1982. Med J Aust, 1985 Jan 21, 142(2), 108 - 11 Genetics and epidemiology of methicillin-resistant Staphylococcus aureus isolated in a Western Australian hospital; Townsend DE et al.; Strains of methicillin-resistant Staphylococcus aureus (MRSA) isolated in the Royal Perth Hospital (RPH) in Western Australia have been analysed genetically and three main types were characterized: (i) strains similar to those isolated in Europe before 1973 . These strains caused small outbreaks in the RPH during the period 1966-1974, but have not been isolated in recent years, except from one patient with reactivation of osteomyelitis after 16 years; (ii) strains of the type prevalent in eastern and northern Australia, one of which caused a difficult-to-control outbreak in the RPH in 1982 . Strains of this type have previously been isolated only from patients who had been in hospitals in eastern and northern Australia, but recently were isolated also from other patients--which indicates that this type of MRSA is now present in the Western Australian community; and (iii) strains, which are genetically different from either of the above two types, were isolated from patients who had been in hospitals in Southeast Asia, but have not yet caused an outbreak in the RPH. Med J Aust, 1985 Jan 21, 142(2), 103 - 8 Control of methicillin-resistant Staphylococcus aureus (MRSA) in an Australian metropolitan teaching hospital complex; Pearman JW et al.; In April 1982, a patient infected with methicillin-resistant Staphylococcus aureus (MRSA) was transferred to the Royal Perth Hospital from the Royal Darwin Hospital . Within three months, 19 patients and four staff members had become infected or colonized with MRSA . The outbreak was terminated only after all colonized inpatients were transferred to a separate isolation unit . After the outbreak, all new patients and new employees who had been in hospitals outside Western Australia in the previous 12 months were screened . From June 1, 1982, to June 30, 1984, 28 of the 649 patients (4.3%) screened on admission to the Royal Perth Hospital were found to be harbouring MRSA . During the same period only one of the 468 persons (0.2%) screened on application for employment at the Hospital was found to be colonized with MRSA . Since the policy of screening new patients and staff from hospitals outside Western Australia was introduced, no serious outbreak of MRSA has occurred. Biochem Biophys Res Commun, 1985 Jan 16, 126(1), 199 - 205 Amino acid sequence of an invertebrate FBP aldolase (from Drosophila melanogaster); Malek AA et al.; The complete amino acid sequence of FBP aldolase from Drosophila melanogaster has been determined . The enzyme contains four identical subunits of 360 amino acid residues . The primary structure of the monomer was established using automated Edman degradation on fragments prepared by CNBr-cleavage, by partial acid cleavage at the unique Asp-Pro bond and by oxidative cleavage at the three tryptophan residues . Manual Edman-Chang degradation was used on smaller peptides obtained by digestion with Staphylococcus aureus V8 protease, trypsin or chymotrypsin . The primary structure of Drosophila aldolase exhibits very extensive homology with the sequence of rabbit muscle aldolase (71% identity), thus explaining the early observation that Drosophila and mammalian aldolases form active interspecies hybrid quaternary structures (Brenner-Holzach, O . and Leuthardt, F., Eur . J . Biochem . (1972) 31, 423-426). Biochem J, 1985 Jan 15, 225(2), 543 - 7 Mannose 6-phosphate-specific receptor is a transmembrane protein with a C-terminal extension oriented towards the cytosol; von Figura K et al.; The portion of the mannose 6-phosphate receptor (nominal Mr 180000 under nonreducing conditions) protruding at the external side of the plasma membrane of fibroblasts and HepG2 cells is susceptible to trypsin . A series of membrane-bound fragments smaller in Mr by 20000-65000 is obtained after incubation of cells with trypsin . When membranes from fibroblasts and HepG2 cells are incubated with trypsin or Staphylococcus aureus proteinase, the receptor is degraded to a single membrane-bound product smaller in Mr by about 9000 . In the presence of 0.1% Triton X-100 extensive degradation of the receptor by trypsin is observed . Furthermore, the receptor in isolated membranes is sensitive to carboxypeptidase Y, which causes a decrease in Mr by about 5000 and 9000 in the absence or presence of detergent, respectively . Mannose 6-phosphate receptor appears to be a transmembrane protein with multiple trypsin-sensitive sites within its larger external (luminal) and smaller C-terminal (cytosolic) portions of the molecule. Virology, 1985 Jan 15, 140(1), 125 - 34 Monoclonal antibodies to the P protein of Sendai virus define its structure and role in transcription; Deshpande KL et al.; Four monoclonal antibodies specific for Sendai virus nucleocapsid protein P were used to examine both the antigenic structure of P and its role in transcription . Three distinct antigenic regions were delineated on P by competitive radioimmunoassays (RIAs), and through Western blot analysis all three sites were mapped to a 40,000-MW (40K) Staphylococcus aureus protease V8-digestion fragment, which remains associated with the neucleocapsid structure . To study the function of P, nucleocapsids were treated with saturating amounts of anti-P monoclonal antibodies and it was found that transcription in vitro was inhibited by 60-90% . Data, therefore, are consistent with the conclusion that the P protein is required for transcription and that the 40K protease-resistant core contains the functionally important portion of the molecule . Further analysis of the P structure showed that some of the 40K fragments were linked by disulfide bonds . These results suggest that the protease-resistant 40K fragment is in the carboxyl-terminal half of P, since the three cysteine residues of P are found there (C . Giorgi, B . M . Blumberg, and D . Kolakofsky (1983), Cell 35, 829-836). J Emerg Med, 1985, 3(3), 227 - 32 Abscess incision and drainage in the emergency department--Part I; Halvorson GD et al.; Superficial abscesses are commonly seen in the emergency department . In most cases, they can be adequately treated by the emergency physician without hospital admission . Treatment consists of surgical drainage with the addition of antibiotics in selected cases . Incision is generally performed using local anesthesia, with intraoperative and postoperative systemic analgesia . Care must be taken to make a surgically appropriate incision that allows adequate drainage without injuring important structures . Postoperative care includes warm soaks, drains or wicks, analgesia, and close follow-up . Antibiotics are usually unnecessary . Complications of incision and drainage include damage to adjacent structures, bacteremic complications, misdiagnosis of such entities as mycotic aneurysms, and spread of infection owing to inadequate drainage . The infectious agents responsible for abscess formation are numerous and depend largely on the anatomic location of the abscess . Staphylococcus aureus accounts for less than half of all cutaneous abscesses . Anaerobic bacteria are common etiologic agents in the perineum and account for the majority of all cutaneous abscesses . Abscesses at specific locations involve special consideration for diagnosis and treatment and may require specialty consultation. Arch Immunol Ther Exp (Warsz), 1985, 33(4), 589 - 94 Phagocytic and bactericidal activities of neutrophils in duodenal ulcer patients during cimetidine treatment; Markiewicz K et al.; Phagocytic and bactericidal activities of neutrophils relative to Staphylococcus aureus 209 P strain were studied in 16 duodenal ulcer patients who were intravenously administered cimetidine in doses of 4 X 200 mg for 8 days and in a group of untreated healthy subjects . The investigations were made before the treatment on the last day of cimetidine administration, and one week after drug withdrawal . The bactericidal activity of neutrophils was found to be higher in duodenal ulcer patients than in the healthy controls . Cimetidine does not have a significant effect on the phagocytic activity of neutrophils and it has a moderately inhibitory effect (p less than 0.05) on the bactericidal activity relative to the ingested intracellular bacteria . This shows that cimetidine may modify some of the neutrophil functions in duodenal ulcer patients. Microbiol Immunol, 1985, 29(8), 709 - 23 Detection of antibody to M protein of measles virus in patients with subacute sclerosing panencephalitis: a comparative study on immunoprecipitation; Ohara Y et al.; Consistent results have not been obtained yet on the presence of antibody to the M protein of measles virus in the sera of patients with subacute sclerosing panencephalitis (SSPE) . We performed a comparative study on various immunoprecipitation systems which appeared in the literature and found that the difference in the composition of the solubilizing buffer produced a large variety of results on the immunoprecipitation . {35S}Methionine-labeled Vero cells infected with the Edmonston strain of measles virus were solubilized by 10 different buffers and reacted with hyperimmune rabbit serum to whole virus, monospecific antisera to H, NP, and M proteins of the virus, normal adults' sera, and the sera from 16 SSPE patients . The immune complex was absorbed by protein A and both solubilization and precipitation rates were compared with each viral protein . Although viral proteins were solubilized by all buffers, the solubilization rate varied considerably . M protein was solubilized and was not coprecipitated nonspecifically with any of the other viral proteins . Purified protein A conjugated to Sepharose was preferable to Staphylococcus aureus for absorption of the immune complex since the latter absorbed both viral and host proteins nonspecifically . The precipitation rates of the viral proteins also varied according to the buffers . Better solubilization of the viral proteins seemed to reduce their rate of precipitation for which the presence of SDS may be responsible, and the presence of the protease inhibitors may also affect the results of immunoprecipitation . Detection of M protein in the immunoprecipitates was largely influenced by the kind of buffer used: some buffers could detect it clearly, but others could not defect it at all . Among the solubilizing buffers tested, Saleh's buffer (Virology 93: 369-376 (1979)),, which contains 0.5% DOC and 0.5% Triton X-100, was most reliable for detection of the anti-M antibody in the rabbit serum, because it showed a high solubilization and high precipitation rates of viral proteins without nonspecific absorption by protein A or coprecipitation of M proteins with any of the other proteins . Using this buffer, we could definitely detect M proteins in the immunoprecipitates from the sera of all six healthy adults and 15 out of 16 patients with SSPE . It was found, however, that the amount of M proteins in SSPE patients was lower than that in healthy adults and varied considerably. Acta Microbiol Hung, 1985, 32(2), 201 - 4 Staphylococcus aureus Tour, a selectively mouse-pathogenic strain for experimental chemotherapeutic study (a note); Uri JV et al.; Staphylococcus aureus Tour is a unique strain . It is highly pathogenic to mice but not to other laboratory animals and primates . This selective pathogenicity makes it useful for experimental chemotherapeutic studies . Since it is highly specialized for mice, it imitates the natural course of infection and produces death after intraperitoneal infection of a relatively few cells even when suspended in isotonic saline . This strain has been found to be very sensitive (MIC's) to various beta-lactams and gentamicin as well as useful for infection-protection studies in mice (ED50's) with a series of cephalosporins. Ultrastruct Pathol, 1985, 8(2-3), 155 - 63 Tubuloreticular inclusions and paired cisternae induced in human lymphocytes cultured with Staphylococcus aureus Cowan 1; Kuyama J et al.; Tubuloreticular inclusions (TRI) and paired cisternae (PC) were induced in lymphocytes of normal individuals after incubation with Staphylococcus aureus Cowan 1 . TRI were initially detected in lymphoid cells on day 2 (48-h culture) . The frequency of TRI-positive cell sections on day 5 increased about twofold over those on days 2-4 . On day 7, TRI were predominantly seen in lymphoplasmacytoid cells or plasmacytoid cells, with an incidence of up to 18% of sections . The regions in these cells were most extensive and anastomosed with the cisternae of adjacent well-developed rough endoplasmic reticulum (RER) . TRI formation appears not to be essential for mitogen-induced B-cell differentiation to plasmacytoid cells, because pokeweed mitogen (PWM) failed to induce TRI . The diverse expressions of TRI induction between these two mitogens may be due to a difference in B-cell activation mechanisms . Paired cisternae were observed in a great majority of mitotic cells at various stages . These were encountered most frequently on day 4 . PC were also seen in the PWM-stimulated culture . Our observations suggest that PC formation may be related to new formation of RER as well as to reconstruction of the nuclear envelope. Scand J Infect Dis, 1985, 17(3), 251 - 7 Serodiagnosis of acute B hepatitis: comparison between a competitive binding radioimmunoassay and an enzyme linked immunosorbent assay for IgM antibody to hepatitis B core antigen; Liaw YF et al.; The standard radioimmunoassay for anti-HBc (CORAB) was modified for the differential detection of anti-HBc IgM by incorporation of a step in which anti-HBc IgG was preferentially absorbed by Staphylococcus aureus cells (Protein A) . The ratio (R) of anti-HBc IgM to total anti-HBc was evaluated by computing the ratio of sample cpm's after and before protein A absorption . The R values of acute B hepatitis ranged from 0.9 to 2.1 (mean 1.3 +/- 0.3) while those of chronic HBsAg carriers ranged from 3.1 to 8.3 (mean 4.9 +/- 1.1) . Adopting 2.1 as the upper limit of R value for acute B infection, this modified CORAB was shown to have excellent correlation with enzyme immunoassay, and to be capable of differentiating acute from persistent HBV infection in HBsAg positive patients, and discriminating acute B hepatitis from non-A, non-B hepatitis in HBsAg negative but anti-HBc positive acute hepatitis. Respiration, 1985, 48(2), 103 - 7 Reactivity of cat tracheal smooth muscle to histamine under conditions of experimentally induced inflammation; Banovcin P et al.; The reactivity of cat tracheal smooth muscle to histamine in vitro was studied at various degrees of inflammation of the airways induced experimentally by the intratracheal administration of turpentine oil or Staphylococcus aureus, both in aerosol form . Tracheal smooth muscle preparations from the control animals did not respond to histamine in doses of 10(-9)-10(-3) mol X 1(-1) . In tracheal preparations from three groups of cats with turpentine oil inflammation induced 24 h, 48 h and 15 days previously, histamine caused contractions in 20, 70 and 24% of the cats, respectively, according to the degree of inflammation . All tracheal preparations from cats with staphylococcal inflammation responded to histamine by contraction . Atropine, acetylosalicylic acid and phentolamine did not abolish histamine contractions in tracheal preparations, but clemastine did. Postgrad Med J, 1985, 61 Suppl 1, 5 - 21 Toxic shock syndrome in Britain--epidemiology and microbiology; de Saxe MJ et al.; By 30 June 1984, only 99 confirmed and probable cases of toxic shock syndrome (TSS) had been reported in the British Isles . Sixty-three were related to menstruation in women aged 14 to 54 years who used tampons of various brands and absorbencies; 33 (52%) of these cases were in girls under 20 . Five women died (8%) and 19 (30%) reported at least one other possible episode . Thirty-six cases associated with a variety of clinical conditions occurred in men aged 17 to 74 years (9), women aged 20 to 54 years (15) and 12 children aged 10 months to 10 years; six patients (17%) died . Strains of Staphylococcus aureus which produced toxic shock syndrome toxin 1 (TSST-1) were isolated from 53 of 58 (91%) menstrual, but only from 18 of 33 (54%) non-menstrual patients . The frequency of toxin production was highest (93%) for 56 vaginal isolates and lowest (33%) for 9 isolates from blood culture . Ninety-six percent (68 of 71) of strains that were TSST-1-positive were sensitive to lytic-group I phages at one of the three concentrations tested; 82% were lysed by phage 29 . Nineteen percent of 339 strains from a variety of sources other than TSS, produced TSST-1, and 35% of the strains lysed by group I phages were positive . Antibody to TSST-1 was detected by enzyme-linked immunosorbent assay at a serum dilution of 1:100, in 232 of 320 (82%) healthy individuals aged 14 to 56 years, but in acute-phase sera from only four of 37 (18%) TSS patients . A rise in antibody levels during convalescence was noted in two menstrual and 5 non-menstrual patients . These results show that the epidemiology of TSS is similar in Britain and the United States and provide further evidence of the importance of TSST-1-producing strains in the aetiology of the disease. Postgrad Med J, 1985, 61 Suppl 1, 39 - 43 Toxic shock syndrome: the effect of solid phase materials on the physiology of Staphylococcus aureus; Holland KT et al.; Rayon and chemically modified cotton were compared for their effects in vitro on the physiology of a strain of Staphylococcus aureus isolated from a patient with toxic shock syndrome . Two types of experiment were carried out . The materials were used to pretreat the medium before culturing in the absence of the materials . Chemically modified cotton was either added or removed from three hour cultures and incubation continued . The rayon treated medium had little effect on growth or exoenzyme/toxin production by S . aureus . The chemically modified cotton greatly reduced growth of S . aureus and decreased exoenzyme/toxin production when added to three hour cultures . However, pretreated medium and cultures grown for three hours with the material present followed by incubation in its absence increased the exoenzyme/toxin production by S . aureus . The in vitro data suggests large effects of chemically modified cotton on the physiology of S . aureus by in some way, at present unknown, altering the growth medium. Infection, 1985, 13 Suppl 1, S9 - 13 Experience with cefotaxime in infections caused by gram-positive pathogens, especially Staphylococcus aureus; Fujii R; Cefotaxime (CTX) was the first third-generation cephalosporin to be launched . According to my classification of cephalosporins for practitioners, in contrast to old beta-lactamase-labile cephalosporins (Group I-III), CTX is beta-lactamase-stable and belongs to Group V with anti-pseudomonas activity . A critical review of about 90 patients with Staphylococcus aureus infections, found among analyzable subjects treated with CTX for gram-positive infections, demonstrates that CTX can be expected to be bacteriologically and clinically effective against this pathogen . Moreover, CTX had excellent efficacy against gram-positive organisms compared with other so-called third-generation cephalosporins . CTX is comparable to or more effective than conventional antibiotics in the treatment of respiratory tract infections, soft tissue infections, and neonatal and pediatric infections caused by gram-positive organisms, S . aureus