Zentralbl Bakteriol Mikrobiol Hyg {A}, 1984 Dec, 258(4), 431 - 40 Purification and properties of succinic oxidase factor, an extracellular product of Staphylococcus aureus; Gemmell CG; Mitochondrial respiration can be inhibited in vitro by a complex of exotoxins elaborated by Staphylococcus aureus . This complex, designated Succinic Oxidase Factor, consists of two components, one of which (A) has a molecular weight of 75,000, is heat-stable, and interferes with cytochrome c oxidase and the other (B) has a molecular weight of 42,000, is heat-labile, and interferes with succinate-cytochrome c reductase . Each component can be separated by Sephadex G200 chromatography and purified by isoelectric focussing . Component A focussed at pH 3.4 and component B focussed at pH 4.5 . Antibody prepared in rabbits to Succinic Oxidase Factor could neutralise the biological activities of each component separately or the whole complex and could react serologically with each component in an agar precipitation test. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1984 Dec, 258(4), 441 - 8 Frequency of pseudocoagulase production in Staphylococcus aureus; Chomarat M et al.; Staphylocoagulase activity could only be separated from that of pseudocoagulase after addition of 4 inhibitors: 3 proteases inhibitors (EDTA, N-ethylmaleimid, aprotinin) and heparin . Of the 32 collection strains studied, only the strains 1503 and NCTC 10 345 (mutant of WOOD 46) exhibited pseudocoagulase activity alone . No other WOOD 46 strain had this isolated activity; on the other hand, 22% of them had simultaneous coagulase and pseudocoagulase activities . One out of 286 Staphylococcus aureus strains of human origin was found to be productive of pseudocoagulase alone, 32% of the strains having both activities . The search for staphylocoagulase coagulating activity in rabbit plasma is a valid test for the identification of Staphylococcus aureus; taxonomical implications are limited without use of a specific detection of coagulase. J Hosp Infect, 1984 Dec, 5(4), 444 - 6 Antibiotic sensitivity and phage typing of Staphylococcus aureus isolated from non-hospitalized patients with angular cheilitis; MacFarlane TW et al.; Strains of Staphylococcus aureus were isolated from 360 patients with angular cheilitis . Of these 24 per cent were sensitive to penicillin G, 74 per cent to tetracycline, 93 per cent to fusidic acid and 96 per cent to erythromycin . Twenty per cent belonged to bacteriophage Group I, 9 per cent to Group II, 13 per cent to Group III, 39 percent miscellaneous and 19 per cent were untypable . A number of phage typing patterns which have been reported for strains associated with specific forms of staphylococcal disease were present in the 360 isolates . In investigations involving cross infection of Staph . aureus, both patients and staff should be examined for evidence of infection at the angles of the mouth. J Hosp Infect, 1984 Dec, 5(4), 417 - 24 Genetic analysis of methicillin-resistant Staphylococcus aureus from a Western Australian hospital; Townsend DE et al.; The isolates from an outbreak of methicillin-resistant Staphylococcus aureus (MRSA) at Royal Perth Hospital (RPH) during a 3 month period in 1982 have been compared genetically with MRSAs isolated at the same hospital during 1969-1973 . The 1982 isolates are genetically similar to isolates from eastern Australia and appear to have been introduced by a patient transferred from a hospital in another Australian state . Methicillin-resistant Staph . aureus isolated during 1969-1973 were genetically distinguishable from the 1982 isolates but were similar to strains reported from elsewhere in the world during these years . This indicates that the MRSA strains currently prevalent in Australia are either a new type or are related to previous MRSAs but have undergone considerable genetic change . These results give further support to suggestions that the strains of MRSA currently being isolated in Australian hospitals have special properties which have facilitated their spread. Thorac Cardiovasc Surg, 1984 Dec, 32(6), 369 - 72 A suspension of fibrin glue and antibiotic for local treatment of mycotic aneurysms in endocarditis--an experimental study; Deyerling W et al.; The finding of mycotic aneurysms creates a major problem in surgery for both active bacterial endocarditis and prosthetic valve endocarditis . The value of local treatment of such aneurysms by a suspension of fibrin glue and an antibiotic was examined in an animal study since a previous in vitro investigation had indicated that such a suspension may discharge sufficient quantities of the antibiotic for up to 12 days . In 3 groups of 6 rabbits each, the entrance to the left atrial appendage was occluded subtotally . The endothelium within the cavity thus created was mechanically injured and the tip of a thin transthoracic catheter was placed in the cavity . In all animals, aliquots of staphylococcus aureus were injected through the catheter . All rabbits developed fever, and positive blood cultures were obtained in 16 . The animals in group 1 were left without treatment . All 6 animals lost weight progressively, 4 animals died from sepsis, 2 rabbits were sacrificed after 6 days . Active endocarditis was demonstrated by histology and bacteriology in each animal . In group 2, 12.5 mg cephalotin were injected via the catheter 24 hours after the infection . Four animals died from sepsis, one rabbit had a positive tissue culture, and only one animal was free of infection on postoperative day 10 . In group 3, 12.5 mg cephalotin suspended in fibrin glue was injected via the catheter 24 hours after the infection . All animals survived, became afebrile and resumed gain of weight . At autopsy after 10 days no infection was detectable . We conclude that a suitable antibiotic suspended in fibrin glue may allow for the sterilization of mycotic aneurysms in bacterial endocarditis. J Biol Chem, 1984 Nov 25, 259(22), 14128 - 35 The 110,000-dalton actin- and calmodulin-binding protein from intestinal brush border is a myosin-like ATPase; Collins JH et al.; A 110-kDa protein present in chicken intestinal brush-border microvilli is believed to laterally link the actin filament bundle that forms the structural core of the microvilli with the microvillar plasma membrane . We have purified a 110-kDa protein to greater than 95% homogeneity by extraction of brush borders with solution containing 0.6 M KCl and 5 mM ATP, followed by gel filtration chromatography, sedimentation as a complex with exogenous actin, and hydroxylapatite chromatography . The 110-kDa protein-calmodulin complex bound F-actin in the absence but not the presence of ATP and had K+,EDTA-ATPase (0.2 mumol/min/mg) and Ca2+-ATPase (0.2 mumol/min/mg) activities and Mg2+-ATPase activity (0.03 mumol/min/mg) that was not activated by F-actin . The actin-binding and ATPase activities of the complex were similar to those of purified brush-border myosin . However, immunoblot analysis showed no reactivity between the 110-kDa protein and polyclonal antibody against purified chicken brush-border myosin . Also, peptide maps of 110-kDa protein and myosin obtained by limited proteolysis with chymotrypsin and Staphylococcus aureus V8 protease had few, if any, peptides in common . Immunoblot analysis also showed that myosin heavy chain was stable under the conditions of the preparation. Biochim Biophys Acta, 1984 Nov 23, 791(1), 102 - 11 Interaction of porcine immunoglobulin M with protein A of Staphylococcus aureus; Gentile TC et al.; Intrigued by reports that the mitogenic effect of protein A on B lymphocytes was due to a direct interaction of cell surface immunoglobulin with protein A, the binding of 19 S, 8 S, and Fab mu fragments of 125I-labeled IgM isolated from porcine serum was investigated . Approx . 60% of purified 19 S porcine IgM interacted specifically with Protein A-Sepharose . Mild reduction and alkylation of 19 S IgM to yield monomeric IgM did not appear to alter its ability to bind to protein A . Elution of either molecular species of IgM from protein A and subsequent repassage over Protein A-Sepharose resulted in nearly quantitative rebinding of the IgM to protein A . Fab mu fragments prepared by digestion of 19 S IgM with pepsin exhibited binding characteristics similar to that observed for intact and monomeric IgM . These results suggest that: (1) there are at least two populations of porcine serum IgM, one that binds to protein A and one that does not; (2) these populations are not interconverting; (3) the ability of IgM to bind to protein A is not dependent on the 19 S pentameric structure extant in sera, but rather is an intrinsic property of some and not all four chain IgM protomers; and (4) a binding site for protein A on porcine IgM is localized in the Fab mu (including the C mu 2 domain) regions of the molecule. J Immunol Methods, 1984 Nov 16, 74(1), 129 - 38 Monoclonal rat anti-Torpedo electroplax nicotinic acetylcholine receptor antibodies: immunochemical characterization; Lukas RJ; The interaction of Torpedo californica nicotinic acetylcholine receptor (nAcChoR) with three rat monoclonal antibodies (mcab) directed against nAcChoR (Gomez et al., 1979) was studied by use of four different radioimmunoassay protocols . Each mcab reacts poorly with formalin-fixed Staphylococcus aureus or S . aureus Protein A, which requires that an additional incubation with second antibody (goat anti-rat immunoglobulin G) is included in each radioimmunoassay paradigm . One mcab inhibits 125I-labeled alpha-bungarotoxin binding to nAcChoR, recognizes a subset of solubilized nAcChoR-toxin complexes, and shows higher titer against nAcChoR in the absence of toxin . Thus, it appears to be directed against nAcChoR antigenic determinants that at least partially overlap with toxin binding sites . Two other mcab react with nAcChoR in a toxin-independent manner . Receptor-mcab dissociation constants are less than 10 nM, according to each assay paradigm . Estimates of antibody titer or concentrations of antibody in stock hybridoma supernatants vary according to the assay used . This predictable result is attributed to differences in design and sensitivity of assay protocols . The data provide a basis for further utilization of monospecific antibodies as probes in characterization of nAcChoR structure and function. J Biol Chem, 1984 Nov 10, 259(21), 13159 - 65 The covalent protein structure of insecticyanin, a blue biliprotein from the hemolymph of the tobacco hornworm, Manduca sexta L; Riley CT et al.; The amino acid sequence has been determined for the insecticyanin from the hemolymph of the fifth instar larvae of the tobacco hornworm, Manduca sexta . The apoprotein is a single polypeptide chain of 189 amino acids, molecular weight 21,378, containing two disulfide bridges, 9-119 and 42-176 . The sequence analysis was performed by automated Edman degradation of reduced and carboxymethylated insecticyanin and fragments generated therefrom by cyanogen bromide, trypsin, chymotrypsin, and Staphylococcus aureus proteinase . Most of the peptides were purified by reverse-phase high-performance liquid chromatography . A purification procedure for the isolation of insecticyanin in high yields and a simple method of determining disulfide linkages are also reported. Proc Natl Acad Sci U S A, 1984 Nov, 81(22), 7127 - 31 Synthesis of a major integral membrane polypeptide of rat liver peroxisomes on free polysomes; Fujiki Y et al.; The manner of synthesis and assembly of the peroxisomal membrane proteins is unknown . Understanding these processes is essential to an understanding of the formation of the organelle . We have investigated the biogenesis of the previously identified major 21.7-kDa integral peroxisomal membrane polypeptide {Fujiki, Y., Fowler, S., Shio, H., Hubbard, A . L . & Lazarow, P . B . (1982) J . Cell Biol . 93, 103-110} . This protein was purified to apparent homogeneity and used to elicit a rabbit antiserum . In immunoblotting analysis, antibody bound only to the 22-kDa membrane polypeptide present exclusively in peroxisomal membranes . Total rat liver RNA was translated in a nuclease-treated rabbit reticulocyte cell-free protein-synthesizing system . The in vitro translation product, isolated by means of the antibody and Staphylococcus aureus cells, comigrated with the mature 22-kDa polypeptide in NaDodSO4/PAGE . Analysis of the translation products of RNAs from free and membrane-bound polysomes indicated that the mRNA for the 22-kDa membrane polypeptide is found predominantly in free polysomes . The results imply post-translational insertion of the membrane polypeptide into the peroxisomal membrane without proteolytic processing and suggest that peroxisomes, like mitochondria and chloroplasts, form by fission from preexisting organelles. J Leukoc Biol, 1984 Nov, 36(5), 633 - 45 Recruitment of neutrophils by an encapsulated coagulase-negative strain of Staphylococcus simulans in the mammary gland of the mouse; Anderson JC et al.; The encapsulated coagulase-negative strain of Staphylococcus simulans (strain 76) was inoculated into the mammary glands of lactating mice . In contrast to the coagulase-positive encapsulated Staphylococcus aureus (strain M), which elicited a neutrophil response within 18 hr, strain 76 organisms multiplied in the glands but failed to elicit a neutrophil response for 3 days; they were then eliminated from the gland by 8 days . When strain 76 was inoculated into mice whose offspring had been removed 3 days earlier, a neutrophil response was induced within 18 hr and, although phagocytic ingestion was resisted for at least 42 hr, the organisms were eliminated from most glands within 5 days . Inoculation of a naturally occurring double-stranded RNA (BRL 5907) into the mammary gland induced a macrophage infiltration of the alveolar lumen that was qualitatively similar to 3 days involution . Intramammary inoculation of strain 76 18 hr after BRL 5907 resulted in a neutrophil infiltration within 6 hr . In vitro, strain 76 stimulated the release of chemotactic factors for neutrophils from bovine mammary gland macrophages . The results suggest that the recruitment of neutrophils by strain 76 in the mammary gland of the mouse is mediated by macrophages. J Clin Microbiol, 1984 Nov, 20(5), 1001 - 2 Prevalence and laboratory identification of methicillin-resistant Staphylococcus aureus in community hospitals; Mahoney MC et al.; The prevalence of methicillin-resistant Staphylococcus aureus infections in community hospitals in northern Minnesota, Wisconsin, and Michigan was found to be one case in 82,565 patients . The percentage of S . aureus isolates resistant to methicillin was less than 0.2% (5 of 2,835) . In this study, conducted from 1 June 1982 to 31 May 1983, a laboratory-controlled methodology was used. Am J Ophthalmol, 1984 Nov, 98(5), 562 - 5 Methicillin-resistant Staphylococcus epidermidis blepharitis; Khan JA et al.; Blepharoconjunctivitis caused by methicillin-resistant strains of Staphylococcus epidermidis occurred in two hospitalized patients, a 74-year-old man and a 19-year-old man . Both strains were resistant to multiple courses of topical antibiotic therapy . Successful treatment in both cases depended upon antibiotic sensitivity testing . These methicillin-resistant strains of S . epidermidis, like methicillin-resistant strains of Staphylococcus aureus, demonstrated almost complete cross-resistance to cephalosporins despite apparent sensitivity on Kirby-Bauer disk sensitivity testing . Topical vancomycin was curative in one case . In the other case, treatment with topical gentamicin, intravenous cefoperazone, and oral rifampin led to resolution of the symptoms. Plasmid, 1984 Nov, 12(3), 197 - 202 Isolation and preliminary characterization of a plasmid mutant derepressed for conjugal transfer in Staphylococcus aureus; Asch DK et al.; The plasmid pCRG1600 is a 52.9-kb self-transmissible plasmid coding for resistance to aminoglycoside and beta-lactam antibiotics in Staphylococcus aureus . When transferred by transduction, plasmid deletion mutants affecting one or more antibiotic-resistance genes were readily obtained . Of these, one derivative (pCRG1690) was found to exhibit a conjugal transfer frequency ca . 100-fold higher than that of the wild-type plasmid . A preliminary physical-genetic map of pCRG1600 located tra in a 14.6-kb region within the 16.9-kb XbaI-A fragment . An 8.5-kb deletion to the left of tra in pCRG1690 was specifically associated with the increased conjugal transferability of the plasmid . Thus, pCRG1690 appears similar to plasmids derepressed for conjugal transfer (drd) in gram-negative bacterial species. J Biochem (Tokyo), 1984 Nov, 96(5), 1525 - 30 A monoclonal antibody to rat liver ornithine decarboxylase; Matsufuji S et al.; A monoclonal antibody was obtained against rat liver ornithine decarboxylase by using hybridoma technology with a small amount of partially purified enzyme . The antibody, IgG1 of kappa-type, was affinity-purified to homogeneity from culture supernatants of hybridoma cells . While the antibody had no inhibitory effect on ornithine decarboxylase activity when tested alone, it precipitated up to 87 units (60 ng) of the enzyme per microgram in the presence of formalin-fixed Staphylococcus aureus Cowan I bacteria . Immunoadsorption on a column of the monoclonal antibody-Sepharose 4B was shown to be useful for the removal of ornithine decarboxylase from antizyme inhibitor preparations, an essential procedure for the accurate assay of either ornithine decarboxylase-antizyme complex or antizyme inhibitor . It was also shown that antizyme could be affinity-purified by using a column of the monoclonal antibody-Affi-Gel 10 to which ornithine decarboxylase had been bound. Rev Infect Dis, 1984 Nov-Dec, 6 Suppl 4, S870 - 4 Failure of a once-daily regimen of cefonicid for treatment of endocarditis due to Staphylococcus aureus; Chambers HF et al.; Cefonicid, a new long-acting cephalosporin, was evaluated for treatment of endocarditis due to Staphylococcus aureus . Four patients, all with infection of the tricuspid valve, were treated with a single daily injection . By the fifth day of therapy, three of the four patients continued to have spiking fevers and positive blood cultures, and treatment with cefonicid was discontinued . Even though peak concentrations of antibiotic in serum were greater than 20-40 times the minimum inhibitory concentration of the antibiotic for the infecting organism, serum bactericidal titers were less than 1:8 in three patients . Susceptibility testing of 52 clinical isolates in broth confirmed a marked difference between inhibitory and bactericidal concentrations for 40% of these strains . In addition, susceptibility testing performed in serum rather than broth resulted in a sixfold increase in the minimum inhibitory concentration, a result suggesting that protein binding may be in part responsible for these failures of treatment . Cefonicid administered as a single daily dose is inadequate for treatment of endocarditis due to S . aureus and should not be used for treatment of bacteremia or life-threatening infections known or suspected to be caused by this organism. Rev Infect Dis, 1984 Nov-Dec, 6 Suppl 4, S829 - 34 Effect of hydrogen ion and protein concentrations on the activity of beta-lactam antibiotics; Platt R et al.; The susceptibility of 50 isolates of Staphylococcus aureus to seven beta-lactam antibiotics was measured under four conditions, involving two pH values and the presence or absence of serum protein . Multiple linear regression analysis was used to determine the effect of pH, protein, and antibiotic on the minimal inhibitory concentrations (MICs) . Each of these factors as well as their interactions had significant effects on the MIC . The effects of pH and protein did not bear a predictable relationship to the extent of binding of antibiotic to serum proteins . All MICs were higher in the presence of protein at both pH values . For some antibiotics, the protein effect at pH 6.0 was larger than that at pH 7.4; for others the protein effect was smaller at pH 6.0 . These data indicate that pH and protein effects must be determined individually for beta-lactam antibiotics. Prikl Biokhim Mikrobiol, 1984 Nov-Dec, 20(6), 798 - 803 {The effect of nitrofurans on the membrane apparatus of coccal bacteria}; Kutsmako RT et al.; Secondary membrane effects on the membrane apparatus of coccus bacteria were being studied . Cultivation of Micrococcus lysodeikticus and Staphylococcus aureus cells on subbacteriostatic concentrations of nitrofurans results in a lower biosynthesis of many membrane proteins, as well as in inhibiting the activity of respiratory enzymes, i . e . the specific concentration of cytochromes and specific activity of NADH-, malate-, lactate oxidases and some reductases drop . Some cytological changes were revealed, when cells were grown on solafur, furazolidone, and furacriline. J Dairy Sci, 1984 Nov, 67(11), 2608 - 13 Evaluation of attenuated, live staphylococcal mastitis vaccine in lactating heifers; Watson DL; Four heifers were immunized in late pregnancy with two doses of attenuated, live Staphylococcus aureus and challenged during early lactation by intramammary infusion into one quarter of approximately 100 organisms of the same attenuated strain . Three unvaccinated control heifers were challenged similarly . At challenge immunoglobulins G1 and G2 antibodies against Staphylococcus aureus surface antigens were significantly greater in blood serum of vaccinated heifers than in controls . Also at challenge, serum from vaccinated heifers had a significantly greater opsonizing capacity for Staphylococcus aureus than did that of controls . The challenge dose of Staphylococcus aureus did not produce prolonged clinical signs of acute mastitis in any of the heifers; however, once of the control animals remained chronically infected . There was a decrease of milk production following challenge for controls but no such decrease for the immunized heifers . Taken together, results of clinical assessments, bacteriology, and measurements of milk production suggested that vaccinated heifers had higher resistance to the challenge dose than did controls. J Dairy Sci, 1984 Nov, 67(11), 2566 - 70 Effect of dietary vitamin A on resistance to experimental Staphylococcus mastitis in mice; Chew BP et al.; Weanling mice (118) were fed a purified diet free of vitamin A for 3 wk and subsequently assigned to diets containing 10, 100 (4000 IU vitamin A/kg diet), or 300% of National Research Council recommended vitamin A . After 3 wk on the treatment diet all mice were bred and allowed to complete gestation . At 24 h postpartum, the left fourth abdominal mammary gland of each mouse was inoculated with Staphylococcus aureus (10(8) cells in .1 ml of saline/gland), and mammary gland infection was observed daily for 6 consecutive days . Liver vitamin A content was lowest in mice for 10% and highest for 300% . However, mice fed 10% showed normal growth and reproduction by small treatment differences in body weight changes, litter size at birth, and average pup weight . Mice fed 10 and 100% vitamin A showed more severe mammary gland inflammation after intramammary inoculation as opposed to mice fed 300% . Severity of mastitis in mice fed 100% vitamin A was similar to 10% . The number of mice classified as mastitic was also similar between 10 and 100% on days 1 and 2 postinoculation; however, on day 3 postinoculation mice fed 100% had a lower incidence of mastitis as opposed to 10% . Severity of mammary inflammation on days 4 through 6 were similar to those on day 3 . Results showed a protective effect of dietary vitamin A supplementation against experimental Staphylococcus aureus mastitis in mice. J Virol Methods, 1984 Nov, 9(3), 227 - 35 Differentiation of Newcastle disease virus strains by one-dimensional peptide mapping; Nagy E et al.; One-dimensional peptide mapping was used for the differentiation of Newcastle disease virus (NDV) strains . Virions were purified in one step, and digested with Staphylococcus aureus V8 protease or chymotrypsin without prior separation of their proteins . Peptides were separated by polyacrylamide gel electrophoresis and stained with Coomassie blue . This method proved to be a simple, economic and reproducible means of differentiating NDV strains. Can J Microbiol, 1984 Nov, 30(11), 1424 - 7 Adaptational changes in Staphylococcus aureus MF 31 grown above its maximum growth temperature when protected by sodium chloride: lipid studies; Hurst A et al.; Staphylococcus aureus MF31 was grown to stationary phase in a complex medium at 30, 37, and 43 degrees C in the absence of salt and at 37 and 46 degrees C in the same medium supplemented with 1 M NaCl . The principal phospholipids were cardiolipin, phosphatidylglycerol, aminoacylphosphatidyl glycerol, mono- and di-glycosyldiglyceride, and traces of phosphoglycolipid . The proportion of cardiolipin decreased with increasing growth temperature, but only slightly in the presence of 1 M NaCl, while that of aminoacylphosphatidyl glycerol was unaffected by growth temperature in absence of salt, but was about halved in the presence of 1 M NaCl . The net negative charge per mole phospholipid was greatly increased in the presence of 1 M NaCl . In the absence of salt, temperature had no effect on the total lipid content, but cells from the 46 degrees C culture in 1 M NaCl contained 25% less total lipid . The proportion of phospholipid in the total lipids, both in the absence and presence of salt, declined with increasing growth temperature . The proportion of glycolipids, however, increased with temperature both in the absence and presence of salt . It is suggested that the increase in glycolipid content and in negative charge/mole phospholipid is a part of the adaptation of S . aureus to the combination of high temperature and 1 M NaCl giving its membrane increased stability and possibly helping to exclude Cl- anion from the cell interior. J Bacteriol, 1984 Nov, 160(2), 792 - 3 Effect of culture pH on the D-alanine ester content of lipoteichoic acid in Staphylococcus aureus; MacArthur AE et al.; The lipoteichoic acid in Staphylococcus aureus growing at high pH values contained very little alanine ester, showing that high overall levels of substitution were not essential for growth . The low alanine content could have resulted from a progressive loss due to base-catalyzed hydrolysis of the labile ester linkages. Infect Immun, 1984 Nov, 46(2), 615 - 8 Primary sequence of the alpha-toxin gene from Staphylococcus aureus wood 46; Gray GS et al.; The complete DNA sequence of a cloned alpha-toxin gene from Staphylococcus aureus was determined . The amino acid sequence of the alpha-toxin protein, predicted from the DNA sequence, was described and compared with published data . The primary product of the cloned alpha-toxin gene contained a 26-amino-acid leader sequence which possessed characteristic features of a signal sequence involved in secretion . The mature alpha-toxin protein had a molecular size of 33,000 and contained only three short regions of high hydrophobicity in addition to a number of short, weakly hydrophobic regions. Infect Immun, 1984 Nov, 46(2), 590 - 7 Presence of toxic shock toxin in toxic shock and other clinical strains of Staphylococcus aureus; Reeves MW et al.; Toxic shock toxin (TST), also known as pyrogenic exotoxin C (Schlievert et al., J . Infect . Dis . 143:509-516, 1981) and staphylococcal enterotoxin F (Bergdoll et al., Lancet i:1017-1021, 1981), was purified from toxic shock strains of Staphylococcus aureus by preparative isoelectric focusing and by chromatofocusing . Neither method produced an absolutely pure protein as determined by silver staining of sodium dodecyl sulfate-acrylamide gels, although chromatofocusing was the better method of the two . Three molecular weight variants of the protein were found in the two toxic shock syndrome strains that were studied, regardless of the purification method that was used . An isoelectric point of 7.15 and molecular weights of 21,400, 22,100, and 23,200 were determined for the different forms of the protein from electrophoresis data . A sedimentation coefficient of 2.3S was determined by sucrose gradient centrifugation, and a Stokes radius of 2 X 10(-7) cm was determined by gel filtration . An average molecular weight of 18,900 for all of the TST forms was calculated from these data by the Stokes-Einstein equation . A survey for TST in 32 control and 46 toxic shock strains of S . aureus by isoelectric focusing and by agarose gel double immunodiffusion with specific rabbit antiserum revealed that the isoelectric focusing method tends to overestimate the number of TST-positive strains because of the detection of non-TST, neutral staphylococcal proteins . Based on immunodiffusion data, the association of TST with toxic shock strains was found to be 100% in vaginal isolates and 62% in non-vaginal isolates . In the control strains, TST was found in 16% of the vaginal strains and 23% of the non-vaginal strains . The value of this toxin as a marker for toxic shock and its relationship to the pathogenesis of this disease are discussed. Infect Immun, 1984 Nov, 46(2), 318 - 23 Correlation between toxin binding and hemolytic activity in membrane damage by staphylococcal alpha-toxin; Bhakdi S et al.; The binding of Staphylococcus aureus alpha-toxin to rabbit and human erythrocytes was studied by hemolytic assays and sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting . Hemolytic assays showed that toxin binding to 10% cell suspensions at neutral pH was very ineffective in the concentration range 3 X 10(-8) to 3 X 10(-7) M (1 to 10 micrograms/ml), and less than 5% of added toxin became cell bound . However, binding was augmented as toxin levels were raised, abruptly increasing to 50 to 60% at 2 X 10(-6) to 3 X 10(-6) M (60 to 100 micrograms/ml) . When rabbit erythrocytes were lysed with 1 to 5 micrograms of toxin per ml, both monomeric and hexameric forms of the toxin could be detected on the membranes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting . In contrast, human erythrocytes treated with 1 to 6 micrograms of toxin per ml did not lyse, and membrane-bound toxin was not detectable . When toxin concentrations were raised to 30 to 100 micrograms/ml, human erythrocytes also lysed and toxin hexamers became membrane bound in comparable amounts as on rabbit cell membranes . Lowering the pH led to a marked increase in susceptibility of human, but not rabbit erythrocytes towards alpha-toxin . When human cells were lysed at pH 5.0 with 5 micrograms of toxin per ml, membrane-bound hexameric toxin became detectable . The demonstrated correlation between the presence of hexameric, cell-bound toxin and hemolytic activity supports the channel concept of toxin-mediated cytolysis . The results also show that toxin binding does not exhibit overall characteristics of a simple receptor-ligand interaction. Infect Immun, 1984 Nov, 46(2), 314 - 7 Toxicity of staphylococcal toxic shock syndrome toxin 1 in rabbits; de Azavedo JC et al.; Strains of Staphylococcus aureus associated with toxic shock syndrome produce toxic shock syndrome toxin 1 (TSST 1), which is lethal to conventional rabbits and acts synergistically with gram-negative lipopolysaccharide . The lethal effect of TSST 1 was examined in specific-pathogen-free rabbits on the basis that these rabbits, being less colonized by gram-negative bacteria, would be less susceptible than conventional animals . Although there was no significant difference in mortality between specific-pathogen-free and conventional rabbits in response to 100 micrograms of TSST 1, there was a difference in response between Dutch belted rabbits and New Zealand white rabbits . Both specific-pathogen-free and conventional New Zealand white rabbits were more susceptible to TSST 1 than the Dutch belted strain . Pretreatment of conventional New Zealand white rabbits with polymyxin B neutralized the lethal effect of TSST 1. Br Heart J, 1984 Nov, 52(5), 591 - 3 Aortic root abscess complicating bacterial endocarditis . Demonstration by computed tomography; Cowan JC et al.; A 68 year old man with an aortic valve prosthesis was admitted to hospital with Staphylococcus aureus endocarditis . Despite antibiotic treatment he continued to be pyrexial . Computed tomography identified a probable abscess between the root of the aorta and the left atrium . The presence of an abscess in this location was subsequently confirmed at operation . Computed tomography is a useful additional diagnostic method for identifying this potentially lethal complication of bacterial endocarditis. Laryngoscope, 1984 Nov, 94(11 Pt 1), 1468 - 71 Microorganisms isolated from infected aural fistulas; Sugita R et al.; Aural fistula is a congenital deformity of the external ear relatively common in Orientals and rare in Caucasians . Suppuration tends to occur, and chemotherapy rather than surgical drainage should be attempted . However, the lack of information concerning infected aural fistulas has made appropriate chemotherapy difficult . Microorganisms isolated from 13 cases of infected aural fistulas were studied from January 1981 to December 1982 . Six species and 22 strains were isolated; nonsporeforming faculative anaerobes were detected in 12 cases . The isolated pathogens included Peptococcus sp (10 cases), Peptostreptococcus sp (3), Bacteroides sp (3), and Fusobacterium sp (2) . One case exhibited only Staphylococcus aureus . Our data also stresses the etiologic importance of anaerobic gram-positive cocci in infected aural fistulas. Mayo Clin Proc, 1984 Nov, 59(11), 785 - 90 Hemorrhagic cardiac tamponade: a clinicopathologic correlation; Olson LJ et al.; Staphylococcus aureus pericarditis and recurrent episodes of hemorrhagic cardiac tamponade developed in a 31-year-old man . He later died of exsanguination and at autopsy was found to have a ruptured infective pseudoaneurysm of the aortic arch . When hemorrhagic pericardial effusions of undetermined cause are encountered, the heart and great vessels should be evaluated as potential sources of the hemorrhage. J Nutr, 1984 Nov, 114(11), 2003 - 9 Increased sensitivity to H2O2 in glutathione peroxidase-deficient rat granulocytes; Baker SS et al.; The role of glutathione peroxidase (GSH-Px) in protecting phagocytic function of peritoneal granulocytes (PMN) was assessed using selenium (Se)-deficient rats . Rats fed an Se-deficient diet for 12-15 weeks developed profound Se deficiency . Their PMN were found to contain 11% of control levels of GSH-Px . Previous studies have shown that at this level of enzyme activity, the metabolism of H2O2 via the glutathione cycle was impaired . Despite this, the initial rate of phagocytosis, as measured by the ingestion of opsonized oil red O particles, was normal . Prior incubation of PMN in an H2O2-generating system resulted in a time-dependent loss in the ability of the cells to ingest . GSH-Px-deficient PMN were affected to a greater degree than control PMN . Degranulation, as measured by the release of beta-glucuronidase into the extracellular medium after stimulation of PMN by opsonized zymosan in the presence of cytochalasin B, was unaffected by GSH-Px deficiency . Prior incubation of PMN in an H2O2 generating system resulted in decreased degranulation in both control and GSH-Px-deficient PMN, with GSH-Px-deficient PMN being affected to a greater degree . The killing of Staphylococcus aureus 502A by both control and GSH-Px-deficient PMN was the same . There was no effect of prior H2O2 incubation on bacterial killing in either control or GSH-Px-deficient PMN . Thus, GSH-Px appears to be important in protecting those aspects of phagocytic function that are sensitive to the destructive properties of exogenous H2O2. J Infect Dis, 1984 Nov, 150(5), 662 - 6 Antibody responses to toxic-shock-syndrome (TSS) toxin by patients with TSS and by healthy staphylococcal carriers; Bonventre PF et al.; Serum samples taken from women with toxic-shock syndrome (TSS) and from women without a history of TSS were examined for the presence of antibodies to toxic-shock-syndrome toxin (TST) . Serum samples from 38 women with TSS and from 70 women with no history of TSS were analyzed by radioimmunoassay (RIA) and by an enzyme-linked immunoadsorbent assay (ELISA) . Antitoxin titers obtained by the assays were highly correlated . Antibody levels in sera of women with TSS, or a history of TSS, were significantly lower than levels in sera of women with no prior evidence of TSS . The mean level of antitoxin titers in the total sample of acute, convalescent, and recovered TSS groups was significantly lower than that of the control groups, which consisted of 31 carriers of genital Staphylococcus aureus and a similar number of age- and race-matched noncarriers . Although a trend toward elevated antitoxin titers was apparent after recovery, no vigorous immunologic response to TST was noted . In contrast, the majority of healthy women demonstrated measurable antitoxin titers, a finding indicative of current or prior colonization with TST-producing strains of S . aureus . The data suggest that absence of antibodies to the TSS toxin may be a predisposing factor in the development of clinical disease. Rev Infect Dis, 1984 Nov-Dec, 6 Suppl 4, S865 - 9 Cefonicid in a once-daily regimen for treatment of osteomyelitis in an ambulatory setting; Kunkel MJ et al.; Fifteen patients with bone joint infections were treated with 1.0 g of cefonicid administered intravenously or intramuscularly once daily . Single organisms isolated included Staphylococcus aureus (from six patients), Staphylococcus epidermidis (three), and Peptococcus species (one) . For four patients infection was polymicrobial, and for one patient no organism was isolated . The mean duration of therapy was 40.4 days, only 10.9 days of which were spent in the hospital . The remainder of therapy was administered intramuscularly in an ambulatory setting . Therapy was successful in all 12 assessable patients . No clinical or bacteriologic relapse occurred in the follow-up period of three to 13 months . The occurrence of adverse effects prompted discontinuation of cefonicid therapy in three patients . Minimum savings in hospital-bed costs alone were $64,350, with 390 hospital days avoided . The minimum savings in work income were $10,010, with 182 days of absenteeism avoided . These data are preliminary but suggest efficacy of cefonicid in a mode of therapy that could have profound cost benefits. Oral Surg Oral Med Oral Pathol, 1984 Nov, 58(5), 545 - 8 The effect of cytotoxic therapy on saliva and oral flora; Main BE et al.; Oral complications of cytotoxic therapy result from direct mucosal damage and, indirectly, occur as a consequence of immunosuppression . Such problems are further exacerbated as a result of associated xerostomia and secondary infection . Therefore, the aims of this study were to examine the salivary volume and composition (amylase, IgA, and lysozyme) together with the oral carriage of potential pathogens in patients receiving cytotoxic therapy . A pilot study comparing healthy controls with patients on chemotherapy for malignant conditions indicated that there were differences between the two groups . Therefore, a longitudinal study was initiated and twelve patients were assessed prior to and 4 and 12 weeks after the start of cytotoxic therapy . The 10-minute forced-spitting salivary volume and amylase and IgA levels all declined significantly over the 12-week period . Lysozyme content did not change . A quantitative increase in the oral carriage of Candida species, coliforms, and Staphylococcus aureus was also observed during therapy . Hence, it is concluded that cytotoxic chemotherapy results in a decreased salivary flow, a reduction in salivary amylase and IgA, and an increase in the oral carriage of opportunistic pathogens. Infect Immun, 1984 Nov, 46(2), 340 - 5 Chemical and biological characterization of a gonococcal growth inhibitor produced by Staphylococcus haemolyticus isolated from urogenital flora; Frenette M et al.; The purified antigonococcal substance produced by Staphylococcus haemolyticus no . 7 has shown a broad antigonococcal spectrum and a narrow antibacterial spectrum . The inhibitor produced in vitro was also active in the guinea pig subcutaneous chamber . The inhibitor has shown hemolytic activity; the human, horse, and mouse erythrocytes were the most susceptible . Hemolytic and antigonococcal activities were inhibited in the presence of phosphatidylcholine . The amino acid composition of the antigonococcal substance was characterized by the absence of proline, tyrosine, histidine, cysteine, and tryptophan . The molecular weight was found to be 2,565, and the major isoelectric points were 4.8 and 4.9 in the presence of 8 M urea and 4.6 without urea . The inhibitor has some properties similar to those of the delta toxin of Staphylococcus aureus, although the two substances are different based mainly on their chemical characteristics . Also an antiserum directed against the gonococcal inhibitor did not give a precipitation line with the delta toxin, indicating that the two substances are antigenically unrelated. Obstet Gynecol, 1984 Nov, 64(5), 666 - 71 Toxic shock syndrome Staphylococcus aureus: effect of tampons on toxic shock syndrome toxin 1 production; Schlievert PM et al.; Tampons were tested for effect on growth and production of toxic shock syndrome toxin 1 by Staphylococcus aureus . Under good growth conditions, regular absorbency tampons had little effect on bacterial growth and inhibited toxin production two- to fourfold . In contrast, higher absorbency tampons had three different effects: 1) some tampons had no effect on bacterial growth but inhibited toxin production; 2) many tampons inhibited both growth and toxin production; 3) one tampon inhibited growth but increased exotoxin per cell . These effects were independent of degree of saturation of the tampons and were observed at incubation times of six, 12, and 18 hours . In no instance was the production of toxic shock syndrome toxin 1 per milliliter increased in the presence of tampons when compared with control. J Immunol, 1984 Nov, 133(5), 2442 - 5 Stimulation of immunoglobulin secretion in human B lymphocytes as a direct effect of high concentrations of IL 2; Ralph P et al.; In certain human IgM and IgG cell lines, immunoglobulin (Ig) secretion is highly stimulated by a B cell inducing factor (BIF) that is free of interleukin 2 (IL 2) . BIF also induces Ig secretion in purified peripheral blood B cell populations that have been mitogenically stimulated by Staphylococcus aureus bacteria . Low concentrations of IL 2 (less than 20 U/ml) are not active in these systems . We now show that IL 2 at concentrations above 100 U/ml can induce Ig secretion in these blood B cells and B cell lines . Both conventional IL 2, purified from the human JURKAT and gibbon MLA-144 cell lines, and recombinant IL 2 are active . Very high concentrations approaching 10(4) U/ml are optimal for Ig secretion . Antibody to the T cell IL 2 receptor, anti-Tac, did not inhibit stimulation of the IgM cell line SKW6.4 by IL 2, and no Tac antigen was detected on the cells . The 9B11 monoclonal anti-IL 2 antibody that neutralizes T cell growth activity also abrogates stimulation of Ig secretion by conventional and recombinant IL 2 in the SKW6.4 cell line . However, the 1H11 monoclonal anti-(conventional thr3-glycosylated IL 2), which does not neutralize T cell growth activity, does inhibit induction of Ig secretion by the corresponding IL 2 in the B cell line . These results suggest that IL 2 stimulates B cells via a low-affinity interaction with a receptor different from the Tac receptor identified on T cells, and that the active site on the IL 2 molecule for B cells differs from that for T cell targets . If IL 2 promotes Ig secretion by binding with a low affinity to the B cell BIF receptor, IL 2 and BIF could be homologous proteins. Dev Biol, 1984 Nov, 106(1), 1 - 14 Formation of the rabbit zona pellucida and its relationship to ovarian follicular development; Wolgemuth DJ et al.; The zona pellucida is the unique extracellular glycoprotein matrix which is assembled during growth of the mammalian oocyte . The present studies were carried out to examine the formation of this structure in relation to the differentiation of ovarian cell types during follicular development . Specific antibodies were developed against total rabbit ZP proteins as well as against ZP proteins electrophoretically purified by high-resolution two-dimensional polyacrylamide electrophoresis gels (2D-PAGE) . Antibodies were characterized by (a) immunoelectrophoresis, (b) a Staphylococcus aureus protein A binding assay, and (c) immunoblotting following 2D-PAGE separation of ZP proteins . Immunoperoxidase localization with these antibodies was used to determine the stage of ovarian follicular development at which ZP antigens first appear as well as to evaluate the cellular and extracellular distribution of these proteins throughout folliculogenesis . The ZP proteins were first observed in the cytoplasm and at the periphery of the oocytes surrounded by a thin squamous follicular cell layer . No staining was observed in the cytoplasm of follicle cells during early folliculogenesis . As the ZP matrix was assembled extracellularly, the intensity of staining of the outer and inner regions could be distinguished . This differentiation of the matrix coincided with the differentiation of the follicular cells into a multilayer cell complex . At this stage, specific ZP proteins are localized within the cytoplasm of the inner layers of these follicular cells . The staining is then diminished in cells of preantral follicles . These studies demonstrate that the formation of the ZP is an excellent model system to study the early stages of follicular development and cell differentiation. Antimicrob Agents Chemother, 1984 Nov, 26(5), 670 - 2 Staphylococcus saprophyticus beta-lactamase production and disk diffusion susceptibility testing for three beta-lactam antimicrobial agents; Latham RH et al.; beta-Lactamase production and MIC determinations for penicillin, methicillin, and cephalothin were assessed for 67 strains of Staphylococcus saprophyticus and correlated with results of disk diffusion susceptibility testing . Fifty-five (82%) of the 67 strains produced beta-lactamase, and 40 (77%) of these beta-lactamase-producing strains were susceptible (zone size, greater than 29 mm) by disk diffusion techniques . Although the range of zone sizes for beta-lactamase producers was broad (26 to 36 mm), all 38 strains with a zone size of less than 31 mm by disk diffusion testing were beta-lactamase producers compared with 17 (59%) of 29 with larger zone sizes (P = 0.0000008) . The median penicillin MIC for 12 S . saprophyticus strains was 0.25 micrograms/ml and was not related to beta-lactamase production . Although the methicillin MICs for 15 strains were in the susceptible range (4.0 micrograms/ml), interpretation of disk diffusion testing for oxacillin varied greatly among laboratories using identically prepared media and standardized techniques . Criteria presently used to define susceptibility of Staphylococcus aureus to penicillin and oxacillin by disk diffusion are inappropriate for S . saprophyticus . The clinical significance of the beta-lactamase produced by these strains needs further evaluation. Proc Natl Acad Sci U S A, 1984 Nov, 81(21), 6827 - 30 Monocytes are required to trigger Ca2+ uptake in the proliferative response of human t lymphocytes to staphylococcus aureus protein A; Lederman HM et al.; We have used the T-cell mitogen Staphylococcus aureus protein A (SpA) to study the role of monocytes in the early events of T-lymphocyte activation . The mitogenic response of human peripheral blood mononuclear cells (PBM) was compared to the response of populations enriched for T cells by E-rosetting (PBM-E+) . In response to SpA, the {3H}thymidine uptake of PBM-E+ was reduced by 80% compared to PBM . The reduced response of PBM-E+ was completely restored by the addition of irradiated PBM-E- or the monocyte-like human cell line U-937 but not by addition of irradiated PBM-E+ . A direct interaction of SpA with monocytes is important since proliferative responses could be generated by preincubation of U-937 with SpA followed by washing and subsequent addition to PBM-E+; incubation of PBM-E+ with SpA followed by washing and subsequent addition of U-937 did not result in a proliferative response . To further delineate the role of the monocyte, we examined the ability of soluble SpA, U-937, or U-937 preincubated with SpA to trigger Ca2+ flux into T lymphocytes, an early step in initiation of the proliferative response . SpA-pretreated U-937, but neither SpA nor monocytes alone, triggered Ca2+ movement into the T lymphocytes . This defines a new role for the monocyte in the early events of T-lymphocyte activation. J Immunol, 1984 Nov, 133(5), 2446 - 53 Regulation of human B cell proliferation by prostaglandin E2; Thompson PA et al.; The role of prostaglandin E2 (PGE2) in the regulation of human B cell proliferation in vitro was examined . Initial studies demonstrated that monocyte (M phi)-mediated suppression of B cell proliferation in Staphylococcus aureus (SA)-stimulated cultures was diminished by indomethacin, and thus it was suggested that cyclooxygenase pathway products of arachidonic acid played a role in the regulation of B cell activation . The possibility that PGE2, one of the major products of this pathway generated by M phi, affected human B cell responses was therefore investigated . PGE2 was found to suppress B cell DNA synthesis and proliferation stimulated by SA in the presence or absence of B cell growth factor (BCGF)-containing T cell supernatants . Suppression was concentration-dependent; significant inhibition was observed in indomethacin-blocked cultures with 10(-10) to 10(-9) M PGE2 . In contrast to the effect of PGE2 on SA-induced B cell proliferation, it had minimal effect on pokeweed mitogen (PWM)-stimulated B cell or T cell DNA synthesis . The effect of PGE2 on the generation of BCGF activity from mitogen-stimulated T cells was also examined . PGE2 (10(-7) M) did not inhibit the production of BCGF activity from PWM-stimulated T cells, or from T cells stimulated with the combination of phytohemagglutinin and phorbol myristate acetate . Thus, PGE2 inhibits B cell proliferation in response to SA but not PWM, and has little effect on the production of BCGF by T cells . These results indicate that PGE2 at physiologically relevant concentrations exerts selective regulatory effects on human B cell responses. Clin Immunol Immunopathol, 1984 Nov, 33(2), 245 - 57 Abnormalities of in vitro immunoglobulin synthesis by peripheral blood lymphocytes from patients with essential mixed cryoglobulinemia; Meroni PL et al.; Peripheral blood mononuclear cells from patients with essential mixed cryoglobulinemia (EMC) were studied for their ability to differentiate into cells containing cytoplasmic immunoglobulins (Ig) and to synthetize Ig after in vitro pokeweed mitogen activation . EMC lymphocytes showed a significant defective differentiation and Ig synthesis compared to normal controls . Coculture experiments carried out mixing enriched normal T- and EMC B-cell suspensions, and vice versa, showed that (a) the EMC B-cell-defective Ig synthesis still persisted after removal of suppressor activity by irradiation, both with autologous and with normal allogeneic T suspensions and (b) EMC T cells displayed a less efficient activity in helping Ig production by normal B lymphocytes . A comparable, reduced response was also found after activation with Staphylococcus aureus strain Cowan I . Taken together these results seem to indicate that in essential mixed cryoglobulinemia an impaired T-cell helper activity coexists with a B-lymphocyte impairment . The significance of these abnormalities in the pathogenesis of EMC is discussed. Arch Biochem Biophys, 1984 Nov 1, 234(2), 341 - 52 Proteolytic dissection of rat brain hexokinase: determination of the cleavage pattern during limited digestion with trypsin; Polakis PG et al.; Limited treatment of rat brain hexokinase (ATP: D-hexose-6-phosphotransferase; EC 2.7.1.1) with trypsin causes cleavage of the Mr 98K enzyme into three major fragments having molecular weights of 10K, 40K, and 50K, with intermediates of Mr 60K and 90K being detected . This information, in conjunction with N- and C-terminal analysis of the intact enzyme and tryptic cleavage products, has established the tryptic cleavage pattern as where T1 and T2 indicate tryptic cleavage sites; cleavage at only T1 or T2 gives rise to the 90K or 60K intermediate, respectively . Confirmation of this cleavage pattern has been provided by two-dimensional peptide mapping using Staphylococcus aureus V8 protease, and epitope mapping with two monoclonal antibodies directed against rat brain hexokinase . The epitopes recognized by one of the monoclonal antibodies is located within the 40K C-terminal fragment while the epitope for the other monoclonal antibody lies within the 50K fragment . A two-dimensional peptide mapping-immunoblotting technique has permitted a more defined localization of these epitopes to specific regions within these major tryptic cleavage fragments . Complete tryptic cleavage of the enzyme occurs with only modest (approximately 20%) loss of catalytic activity, and the cleaved enzyme retains many of the properties of intact hexokinase . Specifically, there was no effect of cleavage on the Km for Glc or the Ki for Glc-6-P, though a slight decrease in Km for ATP was consistently noted to result from cleavage . Furthermore, like the intact enzyme, cleaved hexokinase retained the ability to bind to outer mitochondrial membranes in a Glc-6-P-sensitive manner . Under nondenaturing conditions, the cleaved fragments remain associated by noncovalent forces . Thus, the cleaved enzyme sedimented at a rate comparable to intact enzyme during centrifugation on sucrose density gradients, and migrated only slightly faster when electrophoresed on gradient acrylamide gels under nondenaturing conditions. J Neurochem, 1984 Nov, 43(5), 1352 - 8 Studies on the bovine brain pyruvate dehydrogenase complex using the antibodies against kidney enzyme complex; Sheu KF et al.; Pyruvate dehydrogenase complex (PDHC) was purified from bovine kidney with a specific activity of 12-16 mumol of NADH or acetyl-CoA formed/min/mg protein . The four peptides comprising its three catalytic components were separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) . Rabbit antibodies against this highly purified PDHC (anti-PDHC) exhibited similar binding affinity to the phospho-PDHC as it did to the PDHC antigen . To test whether there exist brain isozymes of PDHC differing from kidney enzyme, which has been extensively characterized, the PDHCs in bovine brain and kidney were compared using this anti-PDHC . The PDHC activities in the brain and kidney mitochondrial extracts were inhibited to the same degree by varying amounts of anti-PDHC . Brain PDHC was precipitated with the anti-PDHC and resolved by SDS-PAGE . The four brain PDHC peptides isolated immunochemically with anti-PDHC had the same sizes as the kidney PDHC peptides . These PDHC peptides from kidney and brain were further compared by their peptide fragment patterns, which were generated by partial proteolysis with Staphylococcus aureus V8 protease or by CNBr and resolved by SDS-PAGE . The peptide patterns generated with the former method indicated that the alpha and beta peptides of the pyruvate dehydrogenase (E1) component and the peptide of dihydrolipoyl transacetylase (E2) component of kidney PDHC were very similar to the corresponding peptides immunologically isolated from brain . The peptide patterns generated with CNBr further confirmed that the beta E1 and E2 peptides of kidney PDHC were similar to the corresponding peptides from brain. Biochem J, 1984 Nov 1, 223(3), 911 - 20 Biosynthesis, transport and processing of myeloperoxidase in the human leukaemic promyelocytic cell line HL-60 and normal marrow cells; Olsson I et al.; The processing and intracellular transport of myeloperoxidase were studied in the human promyelocytic leukaemia cell line HL-60 and in normal marrow cells labelled with {35S}methionine or {14C}leucine . Myeloperoxidase was precipitated with antimyeloperoxidase serum; the immunoprecipitates were subjected to sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and radiolabelled myeloperoxidase visualized by fluorography . During a 1 h pulse, myeloperoxidase was labelled in a chain of apparent Mr 90 000 . With a subsequent chase, the Mr 90 000 polypeptide disappeared and was replaced by chains of Mr 62 000 and 12 400 corresponding roughly to the size of neutrophil myeloperoxidase subunits . The identification of the radioactive polypeptides as different forms of myeloperoxidase was established also by the similarity in patterns generated by partial proteolysis with V8 proteinase from Staphylococcus aureus . Processing of myeloperoxidase in HL-60 was slow; mature polypeptides were significantly increased only after 6 h . Another myeloperoxidase chain of apparent Mr 82 000 was an intermediate precursor or degradation form . Pulse-chase experiments in combination with sucrose-density-gradient separations of homogenates showed that the Mr 90 000 precursor was located in light density organelles only and not in granule fractions, whereas the Mr 82 000 precursor was located only in intermediate density organelles, suggesting that the latter is a product of the former . Processed mature myeloperoxidase was concentrated in the granule fraction, but some occurred in lower density organelles, which may indicate processing during intracellular transport . Only the Mr 90 000 polypeptide was secreted into the culture medium; this was also the only form found in the cytosol fraction. J Bacteriol, 1984 Nov, 160(2), 822 - 3 Simultaneous release of penicilloic acid and phenylacetyl glycine by penicillin-binding proteins 5 and 6 of Escherichia coli; Amanuma H et al.; Penicillin-binding proteins (PBPs) 5 and 6 of Escherichia coli released the bound penicilloyl moiety at an intermediate rate relative to, e.g., Staphylococcus aureus PBPs 4 (rapid) and 1 or 2 (slow) . Each of these E . coli PBPs released the bound penicilloyl moiety as both penicilloic acid (hydrolysis) and phenylacetyl glycine (scission of the C-5--C-6 bond followed by hydrolysis). Res Vet Sci, 1984 Nov, 37(3), 359 - 61 Absence of encapsulation in strains of Staphylococcus aureus isolated from bovine mastitis; Anderson JC; Thirty of 104 strains of Staphylococcus aureus isolated from clinical cases of bovine mastitis in England grew as diffuse colonies in serum soft agar (SSA), 45 grew as mixed diffuse and compact colonies and 29 yielded compact colonies only . The compact strains grew as diffuse colonies in SSA after one passage in the mammary gland of mice . However, none of the strains had an unstained halo when examined by the India ink technique and there was a 99.99 per cent reduction in the viable numbers of the bacteria in 30 representative strains 24 hours after inoculation into the peritoneal cavity of mice . By contrast the truly encapsulated strain M had an unstained halo by the India ink technique and resisted phagocytic killing in the peritoneal cavity . It is concluded that these strains from cases of mastitis are not encapsulated and that growth as diffuse colonies in SSA is not a reliable test of encapsulation. J Biol Chem, 1984 Oct 25, 259(20), 12595 - 601 The primary structure of human liver manganese superoxide dismutase; Barra D et al.; The complete amino acid sequence of manganese superoxide dismutase from human liver was determined . The sequence was deduced following characterization of the peptides obtained from tryptic, chymotryptic, and Staphylococcus aureus digests of the apoprotein . Chemical cleavage with dimethyl sulfoxide-hydrobromic acid was also carried out . The amino acid sequence listed below is made up of 196 amino acids and the two subunit polypeptides in the native enzyme appear to be identical . No homology was observed with copper/zinc containing class of superoxide dismutase . Lys-His-Ser-Leu-Pro-Asp-Leu-Pro-Tyr-Asp-Tyr-Gly-Ala-Leu-Glu-Pro-His-Il e -Asn-Ala-Gln-Ile-Met-Gln-Leu-His-His-Ser-Lys-His-His-Ala-Ala-Tyr-Val-Asn -Asn-Leu-Asn-Val-Thr-Gln-Glu-Lys-Tyr-Gln-Glu-Ala-Leu-Ala-Lys-Gly-Asp-Val -Thr-Ala-Gln-Ile-Ala-Leu-Gln-Pro-Ala-Leu-Lys-Phe-Asn-Gly-Gly-Gly-His-Ile -Asn-His-Ser-Ile-Phe-Trp-Thr-Asn-Leu-Ser-Pro-Asn-Gly-Gly-Gly-Gln-Pro-Lys -Gly-Glu-Leu-Leu-Glu-Ala-Ile-Lys-Arg-Asp-Phe-Gly-Ser-Phe-Asp-Lys-Phe-Lys -Gln-Lys-Leu-Thr-Ala-Ala-Ser-Val-Gly-Val-Gln-Gly-Ser-Gly-Trp-Leu-Gly-Phe -Asn-Lys-Gln-Arg-Gly-His-Leu-Gln-Ile-Ala-Ala-Cys-Pro-Asn-Gln-Asp-Pro-Leu -Gln-Gly-Thr-Thr-Gly-Leu-Ile-Pro-Leu-Leu-Gly-Ile-Asp-Val-Trp-Glu-His-Ala -Tyr-Tyr-Leu-Gln-Tyr-Lys-Asn-Val-Arg-Pro-Asp-Tyr-Leu-Lys-Ala-Ile-Trp-Asn -Val-Ile-Asn-Trp-Glu-Asn-Val-Thr-Glu-Arg-Tyr-Met-Ala-Cys-Lys-Lys. Biochemistry, 1984 Oct 23, 23(22), 5376 - 84 Limited proteolysis of covalently labeled glucocorticoid receptors as a probe of receptor structure; Reichman ME et al.; {3H}Dexamethasone 21-mesylate affinity-labeled glucocorticoid receptors were subjected to controlled proteolysis by trypsin, chymotrypsin, and Staphylococcus aureus V8 protease and then analyzed on denaturing constant percentage or gradient polyacrylamide gels . The molecular weights (Mr congruent to 98 000) and cleavage patterns for rat liver and HTC cell receptors indicated extensive homology between the glucocorticoid receptors from normal rat liver and a transformed rat liver cell line . The major DNA-binding species generated by chymotrypsin treatment was found to be a 42K fragment that was accompanied by several unresolved, slightly lower molecular weight fragments . The meroreceptors obtained after trypsinization were comprised of two species of Mr 30 000 and 28 000 . Each of the three proteases, despite their differing specificities, generated fragments with molecular weights close to 42 500, 30 500, and 27 000 . Nevertheless, each of the three proteases gave rise to a distinctive "ladder" of labeled fragments . No differences could be detected in the digestion patterns of unactivated and activated HTC cell complexes for all three proteases . Also, native and denatured receptor-steroid complexes yielded surprisingly similar digestion patterns with each enzyme . Digestion of denatured complexes readily generated large amounts of a fragment of Mr congruent to 15 000 that was much smaller than the protease-resistant meroreceptors formed from native complexes . The presence of these approximately 15K fragments suggested that the {3H}dexamethasone 21-mesylate labeling of the steroid-binding cavity is restricted to a relatively small segment of the receptor. Br Med J (Clin Res Ed), 1984 Oct 13, 289(6450), 946 - 7 "Normal" acute phase response in systemic sclerosis; Chellingsworth M et al.; An Italian woman with classic active progressive systemic sclerosis had a normal serum concentration of C reactive protein (less than 6 mg/1) . During an infection with Staphylococcus aureus, however, the concentration rose to 250 mg/1 . This was unexpected, since in scleroderma the acute phase response to infection (a brisk rise in serum concentrations of various proteins, including C reactive) has been thought to be defective . This patient is evidence that the acute phase response does occur in systemic sclerosis and that probably it is the nature of the primary disease that masks the response in some cases. J Immunol Methods, 1984 Oct 12, 73(1), 189 - 201 Enrichment of macrophages in cell suspensions of human intestinal mucosa by elutriation centrifugation; Beeken W et al.; To obtain macrophage-rich cell suspensions from human intestinal mucosa, lamina propria specimens were dissociated by incubating in EDTA-collagenase-DNAase solutions and further purified by counter-current centrifugation . During enzymatic incubation macrophage dissociation was linear over the first 8-10 h, reaching a maximum concentration of 10% of total cells and then it plateaued . Counter-current centrifugation resulted in a 5-fold enrichment of macrophages to a mean of 50% with an average recovery rate of 84% . Yields exceeded 0.69 X 10(5) macrophages/g mucosa . Greater than 90% of these cells phagocytosed Staphylococcus aureus, and could be maintained in culture for up to 8 days . Electron microscopy showed satisfactory preservation of the ultrastructure of the macrophages, which also seemed functionally intact. J Biol Chem, 1984 Oct 10, 259(19), 11639 - 42 Conformation-dependent proteolysis of smooth-muscle myosin; Ikebe M et al.; The folded 10 S conformation of turkey gizzard myosin is more resistant to proteolysis by papain than the extended 6 S conformation . These findings confirm those of Onishi and Watanabe (Onishi, H., and Watanabe, S . (1984) J . Biochem . (Tokyo) 95, 899-902) . In addition, we suggest that the effect of phosphorylation on heavy-chain digestion by papain is related to the dependence of conformation on phosphorylation and not to a direct effect of phosphorylation itself . Proteolysis by Staphylococcus aureus protease and trypsin also is slower for 10 S compared to the 6 S conformation . Heavy chain hydrolysis by alpha-chymotrypsin is not dependent on myosin conformation . Filamentous myosin and heavy meromyosin are more resistant to papain proteolysis in the dephosphorylated compared to the phosphorylated states . The different sensitivities to proteolysis probably are caused by changes in the subfragment 1-subfragment 2 region of the molecule rather than at the heavy meromyosin-light meromyosin junction . These changes are induced as part of the 6 S-10 S transition and occur in monomeric and filamentous myosin and in heavy meromyosin . These more subtle alterations in the head-neck junctions of the molecule may be more important in modifying myosin enzymatic activity than the actual interaction of the tail and neck regions of the molecule. J Biol Chem, 1984 Oct 10, 259(19), 12117 - 22 Monoclonal antibodies specific for the alpha subunit of the Escherichia coli DNA polymerase III holoenzyme; Wu YH et al.; We have established three murine hybridoma cell lines that secrete monoclonal antibodies directed specifically against the alpha subunit of the DNA polymerase III holoenzyme of Escherichia coli . All three antibodies have been purified and identified to be of the IgM class . competition binding assays indicate that these antibodies bind to at least two distinct but adjacent or interacting sites . An immunoblot assay has been developed that permits quantitation of alpha in crude extracts . These assays agree with previous determinations of the number of polymerase molecules present/cell and indicate that the size of alpha is the same in both the crude and pure form . Anti-alpha IgM can be used with fixed Staphylococcus aureus, cells and anti-IgM to specifically precipitate alpha. Proc Natl Acad Sci U S A, 1984 Oct, 81(19), 5980 - 4 Localization of a fibrin gamma-chain polymerization site within segment Thr-374 to Glu-396 of human fibrinogen; Horwitz BH et al.; Fibrinogen fragment D1 was converted to fragment D3 by plasmic digestion . This conversion eliminates the ability of the fragment to interact with thrombin-exposed sites on fibrin monomer . Peptides released during this plasmic digestion were assayed for the presence of a polymerization site by affinity chromatography on fibrin monomer-Sepharose . We found that a 33-residue peptide, corresponding to gamma-chain Thr-374 to Lys-406, binds to immobilized fibrin monomer . This peptide is a shorter variant of a previously isolated 38-residue peptide (gamma-chain Thr-374 to Val-411) that contains a polymerization site {Olexa, S . A . & Budzynski, A . Z . (1981) J . Biol . Chem . 256, 3544-3549} . The peptide mixture derived from fragment D1 was digested further with Staphylococcus aureus protease V8, and a 23-residue peptide, gamma-chain Thr-374 to Glu-396, carrying a polymerization site, was isolated by affinity chromatography . This 23-residue peptide inhibits the polymerization of desA-fibrinogen . We conclude that a polymerization site complementary to the site exposed by removal of fibrinopeptide A is present in this segment . The localization of the polymerization site within the gamma-chain segment 374-396 implies that the polymerization site does not overlap with segments of the gamma-chain that are responsible for platelet aggregation and for Staphylococcus clumping (residues 400-411 and 397-411, respectively) or with the residues involved in factor XIIIa-catalyzed fibrin crosslinking (Gln-398 and Lys-406). Appl Environ Microbiol, 1984 Oct, 48(4), 870 - 1 Egg yolk-free Baird-Parker medium for the accelerated enumeration of foodborne Staphylococcus aureus; Lachica RV; A simplified procedure is described for the accelerated enumeration of foodborne Staphylococcus aureus . This involves the replacement of egg yolk in the Baird-Parker medium with Tween 80 and MgCl2 . These compounds, along with pyruvate, allow the recovery of stressed cells of S . aureus on a medium which contains potassium tellurite, LiCl, and glycine as selective agents . Black colonies are identified as S . aureus by the simplified thermonuclease test. Zh Mikrobiol Epidemiol Immunobiol, 1984 Oct, (10), 79 - 82 {Epidemiological assessment of the microbial contamination of the breast milk of healthy women}; Budagovskaia SN et al.; The qualitative and quantitative composition of the microflora of human milk has been studied . The dynamics of the occurrence of different microorganisms constituting the microflora at different periods of lactation has been established . The connection between the appearance of septic purulent infections in infants and the microbial contamination of milk has been revealed . The presence of Staphylococcus aureus in human milk in an amount of 500 microbial cells/ml has been found to constitute no menace to healthy infants. Eur Heart J, 1984 Oct, 5 Suppl C, 97 - 100 Neurologic complications in a group of 86 bacterial endocarditis; Le Cam B et al.; Criteria defined by von Reyn were applied to 86 cases of bacterial endocarditis . Neurologic complications (NC) were categorized according to Pruitt definitions . Neurologic accidents were observed in 48 cases . They were the first clinical manifestation in 20 patients . Neurologic events were of poor prognosis in BE, mortality increasing from 26% in patients without NC to 83% in patients with NC (P less than 0.01) . Two factors affect the incidence of NC: first, the location of endocarditis with 76% of NC in mitral valve endocarditis compared with 37% in other cases (P less than 0.005); and second the infecting organism: 71% of NC in staphylococcus aureus endocarditis versus 45% in endocarditis with other bacteria (P less than 0.02) . Cerebral embolism was the most common NC (25 cases) related to an occlusion of the middle cerebral artery in 21 cases with a fatal outcome in 19 patients . Other NC included 15 intracranial hemorrhages with the evidence of an aneurysm in 4 cases, 6 septic meningitis, 2 macroscopic abscesses, and 2 multiple microscopic abscesses . This study emphasizes the high rate and severity of NC in staphylococcal mitral endocarditis despite antibiotic therapy and supports early surgery in this group of bacterial endocarditis. Arch Microbiol, 1984 Oct, 139(2-3), 240 - 4 Identification of a heat-labile cellular nuclease in Staphylococcus aureus with properties similar to the extracellular nuclease (EC 3.1.4.7); Vakil BV et al.; Besides the well-known heat-stable extracellular staphylococcal nuclease (EC 3.1.4.7) and cell surface bound nuclease, one more nuclease, which is heat-labile, has been identified and purified on phosphorylated cellulose column and characterized . Analyses by Sephadex G-75 gel chromatography indicates that the heat-labile cellular nuclease has molecular weight of about 16,000 similar to those of extracellular and cell-surface bound nucleases . Like the heat-stable nucleases, the heat-labile enzyme acts on both DNA and RNA, is more active on heat-denatured DNA, requires Ca2+ ions for activity and maximum catalytic activity is observed at pH 9.8-10 and at 45 degrees C . The results suggest that the three enzymes have properties strikingly similar to one another and therefore may be related structurally. Antimicrob Agents Chemother, 1984 Oct, 26(4), 563 - 9 Gentamicin uptake in Staphylococcus aureus possessing plasmid-encoded, aminoglycoside-modifying enzymes; Mandel LJ et al.; {3H}gentamicin uptake and killing were studied in three strains of gentamicin-resistant Staphylococcus aureus possessing plasmid-encoded, gentamicin-modifying enzymes and in three isogenic, enzyme-free, gentamicin-susceptible derivatives . At low (less than or equal to 2.0 micrograms/ml) concentrations of gentamicin, uptake by resistant organisms was impaired compared with that of susceptible strains, and no killing was noted . In contrast, at higher (2.5 to 10.0 micrograms/ml) concentrations (which were below the MIC for the resistant strains), rapid gentamicin uptake similar to that seen in susceptible isolates was observed . Although growth inhibition at these concentrations was apparent, there was no loss of viability in resistant strains . Consistently, the membrane H+-ATPase inhibitor N,N'-dicyclohexyl carbodiimide caused resistant strains to take up low concentrations (1.0 microgram/ml) of gentamicin at rates comparable to those seen in susceptible organisms without causing an associated loss of viability . These studies show differences between gentamicin uptake in S . aureus and streptomycin uptake in Escherichia coli (Dickie et al., Antimicrob . Agents Chemother . 14:569-580, 1978) regarding the kinetics of uptake in resistant strains with plasmid-encoded aminoglycoside-modifying enzymes . Specifically, they suggest that for 2-deoxystreptamine compounds such as gentamicin, ribosomal binding followed by accelerated uptake and subsequent interference with cell growth may occur without invariably being associated with lethal effect. Acta Pathol Microbiol Immunol Scand {C}, 1984 Oct, 92(5), 255 - 9 Tumor growth inhibition by protein A and non-protein A containing Staphylococcus aureus in a mouse mammary carcinoma model; Langvad E et al.; Reinfusion of tumor-bearer plasma after absorption with killed Staphylococcus aureus strain Cowan I may be followed by inhibition or even acute necrosis of animal and human tumors . The effect has been attributed to protein A produced in large amounts by this staphylococcus . We have examined the effect upon the growth of a transplanted GR mouse mammary tumor of intraperitoneal inoculation of three strains of S . aureus characterized by being either protein A-rich or protein A-free . A significant tumor growth inhibition was found with all three strains of S . aureus . Serum levels of IgG1, IgG2 and IgM were found to be substantially increased . Crossed immunoelectrophoresis revealed increased numbers and titres of precipitins against staphylococcal antigens . It is concluded that staphylococcal moieties other than protein A may be involved in the tumor growth inhibition . The possibility and role of complement activation by protein A-like molecules through the alternative F(ab)2 reactivity is discussed. Acta Pathol Microbiol Immunol Scand {B}, 1984 Oct, 92(5), 265 - 9 Antibodies to Staphylococcus aureus peptidoglycan and lipoteichoic acid in sera from blood donors and patients with staphylococcal infections; Wergeland H et al.; An enzyme-linked immunoassay (ELISA) was used to detect antibodies in human sera to Staphylococcus aureus peptidoglycan (PG) and lipoteichoic acid (LTA) . All the sera from the blood donors contained IgG antibodies to both substances . Among the sera from 34 patients with bacteriologically verified, serious S . aureus infections, 71 per cent contained significantly elevated levels of anti-PG antibodies and 76 per cent of anti-LTA antibodies . Among the sera from 38 patients with suspected but not bacteriologically verified staphylococcal infections, 58 per cent contained significantly elevated levels of anti-PG antibodies and 74 per cent of anti-LTA antibodies . The levels of antibodies to PG correlated well with the levels of antibodies to LTA, but the latter occurred over a broader range in the patient sera . Elevated antibody values were, however, also found in some patients with serious, non-staphylococcal infections . The diagnostic value of PG and LTA antibodies has to be further investigated. J Pharm Sci, 1984 Oct, 73(10), 1403 - 6 Inhibition of human polymorphonuclear leukocyte cell responses by ibuprofen; Maderazo EG et al.; The stimulation of cell swelling, cell aggregation, polymorphonuclear leukocyte locomotion, and lysosomal enzyme release in response to chemoattractant were all inhibited by ibuprofen, a nonsteroidal anti-inflammatory agent . The dosages needed to induce 50% inhibition (ID50) were 5.9, 7.6, 60, and 95 micrograms/mL, respectively . Aside from the differences in ID50, there was also a difference in the degree of maximum inhibition (Imax) of the complement C5a-stimulated responses observed, so that at achievable serum drug concentrations of 20-50 micrograms/mL, inhibition of 67-78% for cell swelling, 69-82% for cell aggregation, 20-35% for migration response, and 17-38% for lysosomal enzyme release were demonstrated . Also observed were a minor stimulatory effect on nitroblue tetrazolium reduction and an inhibitory effect on the ability to kill Staphylococcus aureus, but only at very high concentrations (approximately 2 mg/mL). EMBO J, 1984 Oct, 3(10), 2399 - 405 Control of pT181 replication I . The pT181 copy control function acts by inhibiting the synthesis of a replication protein; Novick RP et al.; pT181 is a fully sequenced 4.4-kb 20 copy Tcr plasmid from Staphylococcus aureus . Its replication system involves a unique unidirectional origin embedded in the coding sequence for a plasmid-determined protein, RepC, that is required for initiation . When joined to a 55 copy carrier plasmid, pE194, pT181 excludes autonomous isologous replicons by inhibiting their replication . Two types of spontaneous pT181 copy mutants have been isolated, one that eliminates sensitivity to this inhibition and another that does not . A spontaneous 180-bp deletion, delta 144, eliminates both the inhibitory activity and sensitivity to it . This deletion increases copy number by 50-fold and RepC production by at least 10-fold . It is located directly upstream from the repC coding sequence and the deletion-bearing plasmid supports the replication of inhibitor-sensitive plasmids in cells containing active inhibitor . This effect is probably due to the overproduction of RepC by the delta 144 plasmid . On the basis of these results, it is suggested that RepC synthesis is negatively controlled by an inhibitor that is encoded directly upstream from the repC coding sequence and acts as a tareget set in the same region . It is likely, therefore, that pT181 replication rate is determined by the level of RepC. Chem Phys Lipids, 1984 Oct, 35(4), 371 - 84 Preparation and characterization of well defined D-erythro sphingomyelins; Cohen R et al.; A simple semisynthetic procedure for the preparation of various D-erythro sphingomyelins (SPMs), differing in their acyl chains, is described . They were prepared by one-step condensation of the desired free fatty acid with sphingosyl phosphorylcholine (SPC) using dicyclohexylcarbodiimide . The D-erythro SPMs were obtained in high purity, high yields and resemble bovine brain SPM in their chromatographic behavior, infrared, circular dichroism (CD) and proton NMR (PMR) spectra as well as in their rate of hydrolysis by Staphylococcus aureus sphingomyelinase . Multilamellar vesicles can be prepared from the semisynthetic SPMs . Their thermotropic behavior is dependent mainly on the acyl chain though it is also affected by the heterogeneity of the sphingosine base composition . Intact sealed small unilamellar vesicles (SUV) cannot be prepared from a single semisynthetic saturated SPM but can be prepared from their mixtures . This acylation procedure can also be applied for preparing simple neutral glycosphingolipids . The sphingolipids prepared by this method can be used to study metabolism, enzymology and physicochemical properties of D-erythro well defined simple sphingolipids. J Med Microbiol, 1984 Oct, 18(2), 205 - 16 Inhibition of secretion of staphylococcal alpha toxin by cerulenin; Saleh FA et al.; Secretion of alpha toxin by Staphylococcus aureus strain Wood 46 was preferentially inhibited by cerulenin, an antibiotic that stops fatty-acid synthesis by inhibiting beta-keto acyl acyl carrier-protein synthetase . At the concentrations used, cerulenin had a negligible effect on cell growth and total protein synthesis, but reduced lipid synthesis by 50% . Extracellular and membrane-associated alpha toxin was absent in cultures treated with cerulenin, but toxin formation was resumed after either removal of the antibiotic or addition of exogenous fatty acids . The apparent absence of toxin precursor in membranes of inhibited cells favours inhibition at an earlier stage in toxin synthesis. J Med Microbiol, 1984 Oct, 18(2), 197 - 204 Binding of 125I-alpha toxin of Staphylococcus aureus to erythrocytes; Phimister GM et al.; Alpha toxin purified from Staphylococcus aureus strain Wood 46 and radioiodinated by the lactoperoxidase method retained full haemolytic activity and was used to study factors affecting binding to rabbit and horse erythrocytes . A relatively fixed percentage of added toxin bound to both cell types; the percentage bound was independent of temperature, pH, cell concentration and toxin concentration . Neither a 50-fold excess of native toxin nor Concanavalin A inhibited the binding of iodinated toxin to erythrocytes . The results suggest that differences in the sensitivity of erythrocytes to haemolysis do not reflect the abundance of high affinity toxin receptors on sensitive cells, but are more probably the result of differences in the intrinsic stability of the membrane and its sensitivity to perturbation by amphiphilic agents. J Infect Dis, 1984 Oct, 150(4), 583 - 8 Demonstration of differences between strains of Staphylococcus aureus by peptidoglycan fingerprinting; Barzilai A et al.; Lysis of peptidoglycans of Staphylococcus aureus yields products that exhibit strain-specific patterns on analysis by thin-layer chromatography and polyacrylamide gel electrophoresis . The patterns are demonstrated when products of the endogenous autolysins are examined and are enhanced and/or altered by the use of exogenous murolytic enzymes . These fingerprinting techniques were applied to the study of 55 strains of S . aureus . Twenty-six strains were from patients in five localized hospital outbreaks, and 29 were obtained randomly from individual patients . With only two exceptions all strains from a given outbreak had an identical murolytic pattern, whereas the random strains showed variable patterns . Peptidoglycan fingerprinting may be an adjunct tool for the hospital epidemiologist and may also be valuable for the study of certain biologic aspects of interstrain differences. J Infect Dis, 1984 Oct, 150(4), 546 - 53 Foreign body infection: role of fibronectin as a ligand for the adherence of Staphylococcus aureus; Vaudaux P et al.; Foreign bodies made of polymethylmethacrylate coverslips were implanted subcutaneously into guinea pigs, were explanted four weeks later, and were tested for in vitro adherence of Staphylococcus aureus strain Wood 46 . In the presence of serum, the level of staphylococcal adherence to explanted coverslips was 20 times higher than that of adherence to unimplanted coverslips . Adherence to explanted coverslips was caused by fibronectin deposits on the foreign body surface and was inhibited in a dose-related fashion by specific antibodies to fibronectin. J Clin Microbiol, 1984 Oct, 20(4), 806 - 7 Patterns of antibodies to staphylococcal DNases in dog sera; Ness E; Sera from clinically healthy dogs and from dogs with a history of skin disease were examined for the presence of antibodies to the DNases of Staphylococcus aureus, Staphylococcus hyicus subsp . hyicus, and Staphylococcus intermedius . The antibodies found most frequently and in the largest amounts were those to the DNase of S . intermedius. J Clin Microbiol, 1984 Oct, 20(4), 614 - 7 Evaluation of three commercial agglutination tests for the identification of Staphylococcus aureus; Pennell DR et al.; Three commercially available rapid slide agglutination tests for the identification of Staphylococcus aureus were evaluated with 354 recent clinical isolates (165 strains of S . aureus) . The test results of two latex agglutination products, SeroSTAT Staph (Scott Laboratories, Inc.) and Staphylatex (American Micro Scan), and one hemagglutination product, Staphyloslide (BBL Microbiology Systems), were compared with the results of the tube coagulase test, which was read at 4 h (4-h tube coagulase test) and, if negative, again after overnight incubation at room temperature (24-h tube coagulase test) . Discrepancies between agglutination and tube coagulase identifications were resolved by use of the thermonuclease, mannitol fermentation, and slide coagulase tests . All sensitivities, specificities, predictive values of a positive result, and predictive values of a negative result for the three agglutination tests were at least 98.8% and comparable with the 4-h tube coagulase test . Best results were obtained with the 24-h tube coagulase test, which yielded one false-negative and no false-positive tests . Agglutination identifications may be performed on organisms taken directly from a primary plate when sufficient growth is present . Kit agglutination procedures yield rapid and reliable identifications and are easy to perform . This study also demonstrates the usefulness of the 24-h tube coagulase test. J Cell Biol, 1984 Oct, 99(4 Pt 1), 1316 - 23 Cell-specific expression of heat shock proteins in chicken reticulocytes and lymphocytes; Morimoto R et al.; We have found that chicken reticulocytes respond to elevated temperatures by the induction of only one heat shock protein, HSP70, whereas lymphocytes induce the synthesis of all four heat shock proteins (89,000 mol wt, HSP89; 70,000 mol wt, HSP70; 23,000 mol wt, HSP23; and 22,000 mol wt, HSP22) . The synthesis of HSP70 in lymphocytes was rapidly induced by small increases in temperature (2 degrees-3 degrees C) and blocked by preincubation with actinomycin D . Proteins normally translated at control temperatures in reticulocytes or lymphocytes were not efficiently translated after incubation at elevated temperatures . The preferential translation of mRNAs that encode the heat shock proteins paralleled a block in the translation of other cellular proteins . This effect was most prominently observed in reticulocytes where heat shock almost completely repressed alpha- and beta-globin synthesis . HSP70 is one of the major nonglobin proteins in chicken reticulocytes, present in the non-heat-shocked cell at approximately 3 X 10(6) molecules per cell . We compared HSP70 from normal and heat-shocked reticulocytes by two-dimensional gel electrophoresis and by digestion with Staphylococcus aureus V8 protease and found no detectable differences to suggest that the P70 in the normal cell is different from the heat shock-induced protein, HSP70 . P70 separated by isoelectric focusing gel electrophoresis into two major protein spots, an acidic P70A (apparent pl = 5.95) and a basic P70B (apparent pl = 6.2) . We observed a tissue-specific expression of P70A and P70B in lymphocytes and reticulocytes . In lymphocytes, P70A is the major 70,000-mol-wt protein synthesized at normal temperatures whereas only P70B is synthesized at normal temperatures in reticulocytes . Following incubation at elevated temperatures, the synthesis of both HSP70A and HSP70B was rapidly induced in lymphocytes, but synthesis of only HSP70B was induced in reticulocytes. Am J Clin Pathol, 1984 Oct, 82(4), 465 - 9 Clinical and bacteriologic evaluation of BACTEC resin-containing blood culture medium; Callihan DR et al.; In a retrospective study of 1,143 blood culture sets, BACTEC aerobic (6B) and osmotically stabilized (8B) media were compared individually with resin-containing (16B) medium for the isolation of bacteria from the blood of patients receiving antimicrobial therapy . The 16B medium was found to detect significantly more positive cultures than either 6B (P less than 0.01) or 8B (P less than 0.001) . For 22 of 25 isolates of Staphylococcus aureus from 18 patients, 16B medium provided the only means of recovery . All but one of these patients were receiving appropriate antimicrobial therapy, and 16 of 18 had a previous blood culture set positive for S . aureus that did not include a 16B bottle . There was no evidence of a change in antimicrobial therapy in response to a 16B positive culture in these patients . No significant increase in the recovery of other gram-positive or gram-negative bacteria or decrease in the time to radiometric detection of positive cultures as a result of using 16B medium was noted. Clin Pediatr (Phila), 1984 Oct, 23(10), 548 - 52 Childhood osteomyelitis . A five-year analysis of 118 cases in Nigerian children; Okoroma EO et al.; During the years 1976 through 1980, 118 children with osteomyelitis were seen at our hospital, an incidence of almost 24 cases per year . Twenty-eight of these had sickle cell disease . Males were more commonly affected than females, with a ratio of 2.1 to 1, and bones of the lower extremities were more commonly involved, than those of the upper extremities with a 2 to 1 ratio . Seventy patients were anemic, with hemoglobin levels of 10 g/dl or less . Staphylococcus aureus was the most common organism isolated from patients with sickle cell disease, as well as those with normal genotype. J Antimicrob Chemother, 1984 Oct, 14(4), 329 - 38 Presence of two unique genes encoding macrolide-lincosamide-streptogramin resistance in members of the Bacteroides fragilis group as determined by DNA-DNA homology; Callihan DR et al.; The relationship of a previously described, plasmid-encoded macrolide-lincosamide-streptogramin (MLS) resistance determinant to Bacteroides fragilis group clinical isolates from 11 different patients with bacteroides infections was assessed using in situ hybridization . Most of the strains were found to have DNA sequences homologous with the bacteroides MLS plasmid, pBF4, but this homology was exclusively to chromosomal DNA . Two organisms isolated from different sites in a single patient did not share homology with the plasmid, and probably represent a second type of MLS gene . The MLS resistance plasmids, pBF4 (from Bact . fragilis) and pE194 (from Staphylococcus aureus), did not share homologous sequences; therefore at least one of the bacteroids MLS genes is different from that found in Gram-positive organisms. J Antibiot (Tokyo), 1984 Oct, 37(10), 1246 - 52 Pristinamycin accumulation by Staphylococcus aureus; Lacroix P et al.; Pristinamycins IA and IIA (PIA and PIIA) accumulation by Staphylococcus aureus has been studied with two hydrogenated analogs, (H2)PIA and (H2)PIIA . Rapid accumulation of both antibiotics at 37 degrees C is observed and internal concentrations can reach up to 58-fold the external concentration; this accumulation cannot be reduced by either metabolic inhibitors or tetracycline . The synergistic activity of pristinamycins IA and IIA is not observed at the bacterial accumulation level . We propose that pristinamycins enter into bacteria by a passive diffusion process and that the internal concentration is maintained by binding of the antibiotic to the bacterial ribosomes. Int J Oral Surg, 1984 Oct, 13(5), 416 - 22 Characterization of Staphylococcus aureus isolates from oral surgical outpatients compared to isolates from hospitalized and non-hospitalized individuals; Kondell PA et al.; In 160 patients, 114 strains of Staphylococcus aureus were isolated from gingival mucosa, saliva, throat and nose . The patients were divided into 4 groups: one group from an oral surgery clinic for outpatients, one group from a dental clinic, one group from a general surgery clinic for inpatients and one group from a clinic for chronically ill and aged patients . The highest frequency of staphylococcal carriers was found in the outpatient groups (oral surgery and dental clinic), 55% and 45%, respectively . Antibiotic sensitivity testing revealed the majority of the strains to be penicillin resistant, but sensitive to isoxazolyl-penicillins, clindamycin and lincomycin . 50% of the strains produced penicillinase . About 90% of all strains produced lipase, nuclease and a haemolysin most active on rabbit erythrocytes . Human and sheep erythrocytes were lysed by 70% and 46% of the strains, respectively . 77% of the strains were bacteriolytic active . No strain produced lecitinase or elastase . No significant difference was found between the 4 groups in the formation of any of these factors . Phage-type I and III dominated but there was no correlation to enzyme production or patient groups . Thus the 4 patient groups were colonized with strains of Staphylococcus aureus that showed mainly the same pathogenetic factors. EMBO J, 1984 Oct, 3(10), 2407 - 14 Control of pT181 replication II . Mutational analysis; Carleton S et al.; We describe the isolation and analysis of mutations affecting the regulation of Staphylococcus aureus plasmid pT181 replication . Previous results suggested that regulation is achieved by control of the synthesis of RepC, a plasmid-coded replication protein and that the primary negative control element is CopA RNA, which consists of two transcripts that are complementary to the 5' region of the repC mRNA leader . CopA inhibition probably involves a base pairing interaction with the complementary region of the RepC mRNA leader which would facilitate the formation of a downstream stem-loop in the leader that occludes the repC ribosome binding site . RepC is freely diffusible so that regulation of pT181 replication is indirect . Both CopA RNA-sensitive (recessive) and -insensitive (dominant) mutants were isolated . The recessives have defects in CopA RNA structure or activity, the dominants have defects in the site of action (target) of the inhibitor . Some dominants were located within the copA coding sequence . These therefore affect the structure of CopA RNA as well as that of its target . Other dominant mutations mapped outside of the copA gene and therefore produced wild-type CopA RNA . In contrast to directly regulated plasmids, pT181 copy mutants producing wild-type inhibitor could be co-maintained with the wild-type plasmid and mutational changes in inhibitor-target specificity did not change incompatibility specificity. J Oral Maxillofac Surg, 1984 Oct, 42(10), 631 - 6 Osteogenic activity of antibiotic-supplemented bone allografts in the guinea pig; Petri WH 3rd; This study evaluated the osteogenic potential of bone allografts supplemented with up to 1000 times the minimum inhibitory concentration of cephalothin and tobramycin effective against Staphylococcus aureus and Pseudomonas aureus . Mineralized and demineralized bone allografts with and without antibiotics were compared in calvarial defects of guinea pigs . Strontium-85 uptake and histologic evaluation indicated there was no significant difference in osteogeneic activity between nonantibiotic-supplemented allografts and antibiotic-supplemented allografts. FEBS Lett, 1984 Oct 1, 175(2), 267 - 74 A model for the secondary structure of beta-lactamases; Bunster M et al.; A 3-dimensional model, common for the secondary structures of four beta-lactamases obtained from Escherichia coli, Bacillus licheniformis, Bacillus cereus and Staphylococcus aureus, is proposed . The predictions of the structures were made by the hydrophobicity profiles method complemented by the modified Chou and Fasman's method . The model proposed presents 56% constancy and can be described as a 2-domain structure, in agreement with low resolution X-ray data reported for the E . coli enzyme . The model would explain how a common function can be performed by enzymes of very different sizes, composition and sequence. Burns Incl Therm Inj, 1984 Oct, 11(1), 35 - 40 Prospective comparison of silver sulfadiazine 1 per cent plus chlorhexidine digluconate 0.2 per cent (Silvazine) and silver sulfadiazine 1 per cent (Flamazine) as prophylaxis against burn wound infection; Inman RJ et al.; Patients with fresh full-thickness burn wounds were randomly assigned to receive wound treatment with daily applications of either 1 per cent silver sulfadiazine plus 0.2 per cent chlorhexidine digluconate cream (Silvazine) or 1 per cent silver sulfadiazine (Flamazine) . Fifty-four patients treated with Silvazine were comparable to 67 treated with Flamazine with respect to extent and distribution of burn, age and all aspects of wound and associated treatment . Overall incidence of wound bacterial colonization was less in the Silvazine treated patients (65 per cent versus 88 per cent; P = 0.002) . With Silvazine, wound colonization by Staphylococcus aureus was less (41 per cent versus 64 per cent; P = 0.01) . Clinical wound infection with Staph, aureus developed in one Silvazine treated patient and five Flamazine treated patients (P = 0.16) . Colonization by and infection due to all other organisms did not differ in the two groups . The incidence of graft failure was similar with both agents . In future increasing the concentration of chlorhexidine digluconate above 0.2 per cent might produce an improved prophylactic effect against Gram negative bacteria reported by other authors using the combined agent in in vitro and clinical trials . Silvazine was effective in reducing the incidence of Staph . aureus burn wound colonization without fostering supervening opportunistic infection. Scand J Immunol, 1984 Oct, 20(4), 333 - 8 Characteristics of the strongly Ia-positive cells in rat liver; Lautenschlager I; The expression of MHC class II antigens (Ia) on rat liver non-hepatocytic cells were analysed by the Staphylococcus aureus rosette method using monoclonal antiserum to the common part of the class II molecules . Of the non-hepatocytic liver cell population, the passenger lymphocytes and monocytes were strongly anti-Ia binding but the most reactive were large mononuclear cells, morphologically macrophages . The characteristics of the strongly Ia-positive cells of rat liver were tested with different 'macrophage markers' . Of the macrophage-like cells, 55% were Ia-positive in the Staph . aureus rosette 84% had intracytoplasmic lysozyme, 100% of them were positive for alpha-naphthyl acetate esterase (ANAE) reaction, and 92% had Fc-receptors on the cell surface . As functional tests of macrophages, phagocytosis and adhesion were used: 79% of the cells were able to phagocytose antibody-coated human red cells and 56% of the cells adhered on glass . As most of the tested cells were positive for the markers the results demonstrate that the most antigenic component of rat liver has the characteristics of macrophages and thus the strongly Ia-positive cells are probably liver tissue macrophages (Kupffer cells). J Antimicrob Chemother, 1984 Oct, 14(4), 373 - 7 Comparison of ceftazidime, cefuroxime and methicillin in the treatment of Staphylococcus aureus endocarditis in rabbits; McColm AA et al.; Ceftazidime, cefuroxime and methicillin proved equally effective in the therapy of experimental Staphylococcus aureus endocarditis in rabbits with a dosing regimen of 40 mg/kg intramuscularly at 8-hourly intervals for three days . Treated animals all demonstrated a thousand to 10,000-fold reduction in the levels of bacteria in the vegetations compared with untreated controls . In-vitro sensitivities of the organism to the test antibiotics were not predictive of therapeutic efficacy in vivo. J Bone Joint Surg Am, 1984 Oct, 66(8), 1219 - 22 Prophylactic cefamandole in orthopaedic surgery; Gatell JM et al.; Three hundred patients were included in a prospective randomized double-blind trial comparing the efficacy of cefamandole with that of a placebo for prophylaxis of sepsis in operations using Ender or Kuntscher nails, bone plates, or other internal fixation devices . Patients with an open fracture, total joint replacement, or direct operation on the hip were not included in the study . Sixteen patients were excluded because the trial protocol was not followed exactly, so a total of 284 patients participated, 134 of whom were given cefamandole and 150, a placebo . The two groups were similar in terms of mean age, sex ratio, duration of preoperative hospital stay, underlying risk factors, and type of surgical procedure . A superficial wound infection developed in none of the 134 patients who were given cefamandole and in seven of those in the control group (p less than 0.05) . Two deep-wound infections developed in the cefamandole-treated group and four, in the control group (p greater than 0.05) . Staphylococcus aureus, Staphylococcus epidermidis, and gram-negative bacilli were the most common infecting organisms . The rates of infection-related mortality and abscopal infection were similar in both groups . No adverse side effects of the drug were encountered. Eur J Biochem, 1984 Oct 1, 144(1), 101 - 11 The complete amino acid sequence of the major alpha subunit of the lectin from the seeds of Dioclea grandiflora (Mart); Richardson M et al.; The complete amino acid sequence of the major alpha subunit of the lectin from seeds of Dioclea grandiflora was determined . The sequence was deduced from analysis of peptides derived from the native alpha subunit by digestion with trypsin, chymotrypsin, the Staphylococcus aureus V8 protease, and pepsin; and from larger peptides produced by digestion of the citraconylated protein with trypsin . The alpha subunit consists of a single polypeptide chain of 237 amino acids which differs from the sequence of concanavalin in 53 positions . Significant levels of heterogeneity were observed in five positions in the sequence. Clin Immunol Immunopathol, 1984 Oct, 33(1), 99 - 110 Analysis of the mitogenic effects of toxic shock toxin on human peripheral blood mononuclear cells in vitro; Calvano SE et al.; It has been shown previously that the staphylococcal enterotoxins A and B are T-cell mitogens and also cause inhibition of murine plaque-forming cells generated in vitro . Similarly, toxic shock toxin, a 24,000-MW protein produced by toxic shock-associated strains of Staphylococcus aureus, is mitogenic and inhibits the generation of both murine and rabbit plaque-forming cells . In this study, an analysis of the T-cell response to toxic shock toxin was performed . Human peripheral blood mononuclear cells responded to toxic shock toxin over a broad dosage range (1 ng/ml to 5 micrograms/ml) with maximum proliferation at day 4 (96 hr) of culture . Heat treatment (100 degrees C for 60 min) of toxic shock toxin attenuated its mitogenic effects by only a small amount, and this attenuation could be reversed with increasing concentration of the toxin . By cytofluorography, both untreated and toxic shock toxin-treated small lymphocytes manifested normal percentages of OKT3+, OKT11+, OKT4+, OKT8+, HLA/DR+, and Leu-7+ cells . However, toxic shock toxin-induced blasts were 99% OKT11+ and expressed the receptor for interleukin 2 (89%-100% TAC+) . Approximately 85% of the blasts were OKT4+, and 25% of the blasts were OKT8+ . Proliferation of purified, double-rosetted T cells was enhanced monotonically by the addition of irradiated "non-T" cells . Irradiated, monocyte-enriched non-T cells were 2.5 times more potent than unfractionated non-T cells in producing quantitatively similar proliferation by toxic shock toxin-stimulated, autologous T cells . In addition, preincubation of non-T cells for 24 hr with toxic shock toxin, followed by extensive washing and irradiation, induced substantial proliferation by unexposed, autologous T cells . These data show that toxic shock toxin is mitogenic for T cells and requires accessory cells for maximal activity . Further, this substance appears to induce both a subset of OKT4+ (Class II MHC-restricted) and OKT8+ (Class I MHC-restricted) blasts. J Immunol, 1984 Oct, 133(4), 1891 - 5 Response of human B cells to Staphylococcus aureus Cowan I: T-independent proliferation and T-dependent differentiation to immunoglobulin secretion involve subsets separable by rosetting with mouse erythrocytes; Ito S et al.; We separated T-depleted mononuclear cells into subsets by rosetting with mouse erythrocytes and studied proliferation and differentiation responses to Staphylococcus aureus Cowan I (SAC), pokeweed mitogen (PWM), and a combination of the two polyclonal activators . All of the T cell-independent proliferation of unfractionated B cells in response to SAC was attributable to mouse erythrocyte rosette-forming cells (BMR+) . BMR- cells were not stimulated to proliferate by SAC in the presence or absence of T cells, but did proliferate to PWM plus irradiated T cells . Co-stimulation of BMR+ cells with SAC and PWM in the presence of autologous T cells did not lead to immunoglobulin secretion . The B cells stimulated to divide by SAC apparently do not become responsive to B cell differentiation factors and are distinct from those that undergo T cell-dependent differentiation. Cell Immunol, 1984 Oct 1, 88(1), 41 - 8 Human B-cell-stimulatory-factor production by both T4+ and T8+ lymphocytes; Schwarting R et al.; The T-cell subsets responsible for the production of human B-cell-stimulatory factor (BSF) have been identified . Peripheral blood mononuclear cells (PBMNC), E-rosette-forming cells, and isolated T4+/T8+ subpopulations were cultured for 5 days with and without phytohemagglutinin (PHA) stimulation; the supernatants were then assayed for BSF . BSF activity was detected by a costimulation assay using either Staphylococcus aureus Cowan I (SAC) or goat anti-human IgM as a B-cell comitogen . Supernatants from all four PHA-stimulated cell preparations were found to exhibit BSF activity . However, supernatants from the unseparated PBMNC showed a kinetic pattern for BSF production different from that of the E-rosette-enriched T-cell population in that the BSF activity of the former reached a maximum at Day 2, followed by a rapid decrease, whereas BSF production by the latter reached a plateau at Days 4-5 . Although BSF activity was observed in supernatants from both T4+- and T8+-stimulated T-cell subsets, supernatants from T8+ cells contained 50% less activity than supernatants from T4+ cells . Supernatants from the unstimulated fractions and the mitogen-containing medium control did not exhibit BSF activity . These results indicate that both the helper and the suppressor cell fractions, i.e., T4+ and T8+ lymphocytes, are responsible for the production of BSF . However, it is unclear whether the BSF activities detected in both T-cell subsets are mediated by the same or different molecular entities. J Neurochem, 1984 Oct, 43(4), 935 - 43 Cross-reaction of anti-rat B-50: characterization and isolation of a "B-50 phosphoprotein" from bovine brain; Oestreicher AB et al.; Antibodies to the phosphoprotein B-50 of rat brain were used to trace cross-reacting brain proteins of vertebrates . With the SDS-gel-immunoperoxidase method, a cross-reacting protein (CP) of apparent Mr 53,000 was demonstrated in the homogenate and the synaptic plasma membrane fraction of bovine brain . Sequence 1-24 of adrenocorticotropin (ACTH1-24) (10(-5) M and 10(-4) M) inhibited endogenous phosphorylation of CP in synaptic plasma membranes . The protein was partially characterized and purified to homogeneity from bovine brain by procedures previously described for rat B-50 . CP was enriched in ammonium sulfate precipitated protein (ASP) fractions and phosphorylated by an endogenous protein kinase . Two-dimensional gel analysis of bovine and rat ASP showed that the cross-reacting protein had an isoelectric point less acidic than B-50 . Limited proteolysis by Staphylococcus aureus protease yielded a "peptide map" analogous to B-50 . Two major fragments of Mr 30,000 and 17,000 were produced . In addition, CP exhibited other similarities to rat B-50: phosphorylation by rat brain protein kinase C, microheterogeneity observed after isoelectric focusing, and possibly degradation by endogenous proteolysis . Cross-reaction of proteins in brain homogenates of other mammalian species and of chicken was demonstrated: the Mr of the proteins ranged from 47,000 to 53,000 . We conclude that (1) the cross-reacting bovine protein is a "B-50 protein," and (2) the Mr of the "B-50 protein" varies from species to species. Biull Eksp Biol Med, 1984 Oct, 98(10), 499 - 502 {RNA synthesis in the polymorphonuclear leukocytes of a suppurative wound}; Sarkisov LS et al.; RNA synthesis in biopsy specimens of burn or traumatic wounds and in blood leukocytes was investigated in 11 patients by electron microscopic radioautography . Experiments with leukocytes were carried out in two variants: 1) incubation with 3H-uridine; 2) incubation with 3H-uridine in the presence of Staphylococcus aureus of Staphylococcus epidermidis suspensions . It was detected that neutrophils incubated without bacteria did not contain the label . Neutrophils incubated with bacteria and phagocytizing them could incorporate 3H-uridine . The label was insignificant in this case and the number of labeled cells did not exceed 18% . The majority of neutrophils (70-90%) in pus accumulations from burn and traumatic wounds contained the label in a higher concentration than neutrophils that phagocytize bacteria in test-tubes. Proc Natl Acad Sci U S A, 1984 Oct, 81(20), 6471 - 5 Remission of leukemia and loss of feline leukemia virus in cats injected with Staphylococcus protein A: association with increased circulating interferon and complement-dependent cytotoxic antibody; Liu WT et al.; We have injected purified Staphylococcus aureus protein A intraperitoneally into leukemic cats infected with feline leukemia virus, into cats persistently infected with feline leukemia virus but without hematologic or cytologic abnormalities, and into healthy cats without feline leukemia virus infection . Pre- and post-treatment serum samples were evaluated sequentially for interferon activity and for complement-dependent cytotoxic antibody . Our results indicate that serum interferon increased dramatically (less than 3 to 324 units/ml) during treatment only in cats that responded to staphylococcal protein A therapy . Increase of interferon preceded or was closely associated with increasing levels of cytotoxic antibody, loss of viremia, and correction of cytological and hematological abnormalities of three leukemic cats . The cytotoxic antibody was shown to be specific for envelope glycoprotein gp70 of the feline leukemia virus . One persistently feline leukemia virus-infected cat without leukemia that became nonviremic also developed high levels of interferon and specific cytotoxic antibody . By contrast, interferon levels of cats not responding to treatment had levels of less than 3 to 27 units/ml . Normal healthy cats injected with staphylococcal protein A showed moderate transient increases of interferon but no detectable cytotoxic antibodies