Antimicrob Agents Chemother, 1984 May, 25(5), 575 - 8 Tolerance percentage as a criterion for the detection of tolerant Staphylococcus aureus strains; Goessens WH et al.; In this study, the degree of tolerance was determined in several populations of Staphylococcus aureus isolates . The degree of tolerance of a staphylococcal strain can be established in a reproducible way by exposing the strain to increasing concentrations of a beta-lactam antibiotic and determining the number of surviving bacteria at each concentration . The number of surviving bacteria was expressed as a fraction of the initial inoculum . By this technique, it appears that for each strain the value of the surviving fraction stabilized above a certain concentration of the antibiotic . This value was called the tolerance percentage of the strain . In 64 S . aureus strains isolated from blood cultures in 1982, the tolerance percentages, after exposure to methicillin, varied from less than or equal to 0.1 to 6; 28% of the strains showed a tolerance percentage of less than or equal to 0.1, and 12.5% showed a tolerance percentage of greater than or equal to 2 . Similar tolerance percentages were found with cloxacillin, nafcillin, cephalothin, and penicillin . Strains with a tolerance percentage of greater than or equal to 2 showed slow killing and lysis in the presence of a high methicillin concentration . A tolerance percentage of 2 appeared to be the breakpoint between susceptible and tolerant strains . Older collections of S . aureus strains, dating from the years 1951 to 1953 and 1957 to 1958, also included strains with a survival percentage of greater than or equal to 2, thus indicating that tolerance of S . aureus to beta-lactam antibiotics is not a new phenomenon. Am J Med Sci, 1984 May-Jun, 287(3), 49 - 54 Rifampin in the treatment of serious staphylococcal infections; Swanberg L et al.; Eight patients with serious staphylococcal infections such as endocarditis, meningitis, epidural abscess, shunt and graft infections were treated with nafcillin, cephalothin or vancomycin in combination with rifampin . In vitro antibiotic sensitivities demonstrated that the Staphylococcus aureus and Staphylococcus epidermidis were highly sensitive to rifampin with minimum inhibitory and bactericidal levels of .0078-.25 micrograms/ml . In all patients reviewed and reported, when serum bactericidal levels were measured before and after the addition of rifampin, there was a positive correlation between microbiological and clinical outcome . Thus, in selected patients with life-threatening infections caused by S . aureus and S . epidermidis rifampin should be considered an adjunctive therapy. Arch Fr Pediatr, 1984 May, 41(5), 353 - 5 {Chronic septic granulomatosis . Disclosed by hepatic abscesses . Contribution of echography}; Cheron G et al.; Skin infections and lymph node abscesses are the most common manifestations of chronic granulomatous disease (CGD) during the first year of life . Intra-hepatic abscesses occur later on . The case reported concerns a 4 month-old infant, in whom the disease presented initially with liver abscesses . Percutaneous needle aspiration of the liver under ultrasonographic control permitted the isolation of Staphylococcus aureus and allowed partial regression of the abscesses . Surgery was not required. Pediatr Infect Dis, 1984 May-Jun, 3(3), 222 - 5 Detection of teichoic acid antibodies in children with staphylococcal infections; Thisyakorn U et al.; The presence of serum antibodies to teichoic acid was evaluated by gel diffusion and enzyme-linked immunosorbent assay in 14 patients with deep-seated staphylococcal infection, in 5 patients with superficial staphylococcal infections, in 10 patients with Gram-positive infections other than staphylococcal and in 12 age-matched, uninfected patients . Serum samples were obtained on admission and serially each week during hospitalization . Teichoic acid antibodies were detected by gel diffusion in only 5 of 14 patients with deep-seated staphylococcal infections, in 1 of 10 patients with other Gram-positive infections and in none of the other patients . With the enzyme-linked immunosorbent assay method all patients with deep-seated staphylococcal infections had concentrations of teichoic acid antibodies of 1:1600 or greater, and these titers were significantly larger than those in the other groups of patients . Using a titer of 1:3200 or greater as a diagnostic level in children with deep-seated Staphylococcus aureus infections, the sensitivity was 93% and the specificity was 89% . For all staphylococcal infections the sensitivity was 79% and the specificity was 96%. Pathol Biol (Paris), 1984 May, 32(5), 362 - 8 {Resistance to pristinamycin (or virginiamycin) of strains of Staphylococcus aureus}; El Solh N et al.; Resistance to pristinamycin (or virginiamycin) was first encountered in Staphylococcus aureus strains in 1975 . These strains are usually multiresistant, in particular to streptogramin A components (SgA), macrolides, lincosamides and streptogramin B components (ML SgB ) . Results of molecular analysis of 16 such strains, recently isolated, suggests that SgA resistance is not encoded by plasmid genes . Curing and mixed culture experiments allowed us to dissociate SgA from SgB resistance genes . Conversely, in a previous study on other strains, the same two resistance genes were shown to be carried by a single plasmid and could not be dissociated . Since 1981, a new type of pristinamycin -resistant S . aureus strains has been isolated . These strains are resistant to SgA and lincosamides but susceptible to macrolides and SgB . Eight such strains from 3 parisian hospitals have been studied . In mixed culture experiments, SgA resistance and penicillinase genes always transferred jointly . In some instances, these two determinants also cotransferred with genes encoding lincomycin, lincomycin and clindamycin, and/or aminoglycosides resistance. J Infect, 1984 May, 8(3), 221 - 6 Comparison of anti-alpha-haemolysin and teichoic acid antibody tests in patients with Staphylococcus aureus endocarditis or bacteraemia; Larinkari U et al.; To assess the value of serological tests in differentiating endocarditis and complicated bacteraemia due to Staphylococcus aureus from uncomplicated S . aureus bacteraemia and from nonstaphylococcal endocarditis we measured teichoic acid antibodies (TAA) (by means of a gel diffusion method) and antibodies to alpha-haemolysin (ASta) in the serum of 22 patients with S . aureus endocarditis, 42 patients with complicated S . aureus bacteraemia, 21 patients with uncomplicated S . aureus bacteraemia, 27 patients with other than S . aureus endocarditis, 17 patients with culture-negative endocarditis and 337 non-infected control patients . TAA and ASta titres were found significantly more often in staphylococcal endocarditis than in non-staphylococcal endocarditis, 59 per cent versus 4 per cent for the TAA test and 32 per cent versus 0 per cent for the ASta assay . The combined use of the two tests proved best for differentiating the two groups of endocarditis from each other, 72 per cent versus 4 per cent respectively . In the culture-negative endocarditis group there were two serologically positive patients whose anti-staphylococcal antibiotic therapy based on the serological findings was successful, supporting the clinical usefulness of staphylococcal serological tests in endocarditis of unknown bacterial aetiology . The serological tests were not useful, however, in differentiating S . aureus endocarditis from complicated or uncomplicated bacteraemia due to S . aureus without endocarditis. Mol Pharmacol, 1984 May, 25(3), 494 - 501 Purification of liver microsomal cytochrome P-450 isozymes 3a and 6 from imidazole-treated rabbits . Evidence for the identity of isozyme 3a with the form obtained by ethanol treatment; Koop DR et al.; Two forms of cytochrome P-450 have been purified to electrophoretic homogeneity from hepatic microsomes of rabbits treated with imidazole . Several criteria indicate that the cytochrome of higher electrophoretic mobility is identical with ethanol-inducible isozyme 3a . "Imidazole-3a" and "ethanol-3a" exhibit the same chromatographic characteristics and have identical electrophoretic mobilities upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Furthermore, the two protein preparations have the same absorbance maxima and absorption coefficients in the oxidized, reduced, and reduced-CO states . A single immunoprecipitin band exhibiting complete identity was observed upon reaction of imidazole-3a and ethanol-3a with the immunoglobulin G fraction from sheep immunized with the latter protein . The amino acid composition and first 10 residues of the amino terminus of the two protein preparations are indistinguishable, as are the high-performance liquid chromatographic maps of the peptides obtained upon cleavage with trypsin, Staphylococcus aureus V8 protease, or Lys C endoproteinase . Furthermore, these preparations have very similar activities in the oxidation of ethanol to acetaldehyde and the p-hydroxylation of aniline . Evidence was obtained that the cytochrome of lower electrophoretic mobility isolated from imidazole-treated rabbits is probably identical with isozyme 6; the spectral characteristics, amino acid composition, and carboxyl-terminal sequence are described . As an inducer, imidazole has the advantage over ethanol of being less variable in its effects and requiring a shorter period of treatment . From the resulting liver microsomes, one can readily isolate, in addition to P-450 isozymes 3a and 6, isozymes 3c and 4 as well as epoxide hydrolase. Prikl Biokhim Mikrobiol, 1984 May-Jun, 20(3), 363 - 8 {Isolation and properties of an extracellular proteinase of Staphylococcus aureus}; Beliaeva EV et al.; Serine proteinase splitting the peptide bonds which are formed by carboxyl groups of dicarboxylic amino acids was isolated from the supernatant of the Staphylococcus aureus culture liquid . It is similar to the enzyme isolated by Drapeau from Staphylococcus aureus strain V8 in its specifidity, molecular weight, amino acid composition, existence of two pH optima (pH 4.6 and 8.2) . But there are some differences between the two proteinase in the content of dicarbonic amino acid residues . It was found that the enzyme can exist in two molecular forms. Appl Environ Microbiol, 1984 May, 47(5), 965 - 70 Comparison of methods for recovery of Escherichia coli and Staphylococcus aureus from seeded laundry fabrics; Cody HJ et al.; To assess the effect of laundry procedures on fabric-associated bacteria, a standard method of enumeration is needed . We evaluated six methods for enumeration of Escherichia coli and Staphylococcus aureus seeded (10(2) and 10(5) CFU/100 cm2 of fabric area) onto sterilized hospital sheets and terry . Two methods involved maceration of seeded swatches in broth followed by passage of the broth through a 0.45-micron-pore-size, 47-mm-diameter filter membrane . Three methods involved agitation of seeded swatches in broth with a paint shaker and membrane filtration of the broth to recover eluted bacterial cells, and the final method involved direct enumeration of cells on fabrics by overlaying seeded swatches with agar containing triphenyltetrazolium chloride as an indicator . The most convenient recovery method employed a 90-s agitation followed by serial dilution of broths and membrane filtration . This method provided 44/57% (low seed/high seed) recovery of E . coli from sheets and 133/31% from terry and 34/74% recovery of S . aureus from sheets and 58/57% from terry . Although maceration provided similar recovery of E . coli and S . aureus, it is a less-practical method . The direct enumeration method was ineffective for enumerating gram-positive bacteria . We conclude that either the agitation or maceration method used enumerated the seeded bacteria to within 1 log10 of their expected number and can be used to assess the bactericidal effectiveness of various steps in the laundering process. J Clin Microbiol, 1984 May, 19(5), 680 - 6 Methodological aspects of Staphylococcus aureus peptidoglycan serology: comparisons between solid-phase radioimmunoassay and enzyme-linked immunosorbent assay; Christensson B et al.; In the present studies we compared the ability of two commonly used assays, solid-phase radioimmunoassay and enzyme-linked immunosorbent assay (ELISA), to detect human antibodies to Staphylococcus aureus peptidoglycan . ELISA was superior, with a reproducibility of 12.0%, as compared with 18.1% in solid-phase radioimmunoassay . Much lower serum dilutions could be used in ELISA . We also studied the effects of solubilizing the antigen by lysostaphin, lysozyme, or ultrasonication . Lysostaphin-treated peptidoglycan cannot be recommended since solid-phase radioimmunoassay could not distinguish positive from negative serum samples with this preparation . On the other hand, the sensitivity in both assays was high when peptidoglycan treated with lysozyme for 240 min or with ultrasonication for 30 min was used as antigen . The interassay correlation between solid-phase radioimmunoassay and ELISA was slightly better with sonicated peptidoglycan (correlation coefficient = 0.94, P less than 0.01), as compared with lysozyme-treated peptidoglycan (correlation coefficient = 0.76, P less than 0.01) . We recommend the ELISA with sonicated peptidoglycan as antigen for use in routine serology. Clin Exp Immunol, 1984 May, 56(2), 415 - 24 Cord blood B cell differentiation . Synergistic effect of pokeweed mitogen and Staphylococcus aureus on in vitro differentiation of B cells from human neonates; Miller KM et al.; B lymphocyte differentiation into immunoglobulin secreting cells is a process depending on the presence of functionally mature B lymphocytes, monocytes and regulatory T lymphocytes . Cord blood B lymphocytes present in isolated cord blood mononuclear cell (MNC) preparations are normally unable to differentiate into immunoglobulin secreting plaque forming cells (PFC) when cultured in the presence of pokeweed mitogen (PWM) alone or killed Staphylococcus aureus Cowan 1 (SAC1) alone . However, each one of these activators induces PFC formation by B lymphocytes in adult MNC cultures . In the present study we show that these two activators can act synergistically to induce a significant in vitro PFC response in cord blood MNC's . The synergism of PWM and SAC1 exhibits a requirement for a specific sequence of addition in order to induce a positive response in neonatal cells . If both activators are not added simultaneously at the initiation of culture, only the initial addition of SAC1 followed by PWM will result in increased PFC production . The action of PWM and SAC1 on cord blood MNC can each be replaced by conditioned media . Supernatants from monocyte cultures containing soluble factors such as interleukin-1 (IL-1) can substitute for the activity of SAC1 while supernatants containing soluble T-cell factors (interleukin-2{IL-2}, T cell replacing factor (TRF), B cell differentiation factor (BCDF), etc) can replace PWM in the cord blood MNC cultures . The results suggest that the synergistic effect of these two activators overcomes a partial immaturity or an excessive suppressor activity of human cord blood MNC. J Clin Immunol, 1984 May, 4(3), 220 - 7 Chronic granulomatous disease in two sisters; D'Amelio R et al.; Two sisters with chronic granulomatous disease (CGD) have been studied . The diagnosis was suggested by the histopathological findings from the spleen and lymph nodes of the proband and confirmed by the low values obtained in the following tests performed on polymorphonuclear leukocytes (PMN): chemiluminescence, nitroblue tetrazolium (NBT) reduction, killing of Staphylococcus aureus, and O2- production . NADPH oxidase activity was not detected in the homogenates of the patients' PMN but cytochrome b was normally present . In addition, PMN depolarization induced by phorbol-myristate acetate was absent, thus suggesting a defect of the activation mechanism of the respiratory enzyme . The normal depolarization induced by ouabain indicated that the membrane polarity regulated by the Na/K pump in the patients' cells was not affected . The low, but not completely absent, respiratory activity of the patients' PMN could suggest an X-linked mode of inheritance with incomplete Lyonization . From a clinical point of view, one sister had mild symptoms whereas the other was almost symptomless, thus confirming once more the heterogeneity of CGD syndrome. Infect Immun, 1984 May, 44(2), 465 - 8 Inhibition of cell-free oxidative bactericidal activity by erythrocytes and hemoglobin; Hand WL; Sickle cell anemia and other chronic hemolytic anemias are associated with an increased frequency of bacterial infections . There is evidence to suggest that in hemolytic states massive erythrocyte (RBC) ingestion by macrophages interferes with their antibacterial function, thereby predisposing infection . Stimulated by this possibility, we recently demonstrated that erythrophagocytosis by macrophages markedly inhibited intracellular killing of bacteria, and that zymosan-stimulated superoxide generation and chemiluminescence were also suppressed by RBC ingestion . We examined the effects of RBC components on generation of chemiluminescence, superoxide, and bactericidal activity by cell-free oxidative systems . Generation of chemiluminescence by hypoxanthine-xanthine oxidase was depressed in the presence of human RBC lysate or column-fractionated hemoglobin but not crystallized human hemoglobin (methemoglobin) (peak cpms of 15,522 {P = 0.00024}, 28,360 {P = 0.0088}, and 50,041 {P = 0.37}, respectively, compared with 59,898 for positive controls) . Similarly, hypoxanthine-xanthine oxidase production of superoxide was inhibited in the presence of column-fractionated human hemoglobin (43.8 versus 17.4 nmol per tube, P = 0.000001) . A cell-free bactericidal system, acetaldehyde and xanthine oxidase with or without myeloperoxidase and Cl-, was markedly inhibited by column-purified hemoglobin . For example, after 2 h of incubation, surviving numbers of Staphylococcus aureus were: control (buffer only), 2.5 X 10(6)/ml; bactericidal system, none; bactericidal system plus hemoglobin, 2.2 X 10(6)/ml (P less than or equal to 0.03, bactericidal system versus other systems) . Our studies have documented that interactions between RBC (hemoglobin) and reactive products of oxygen metabolism inhibit oxidative bactericidal mechanisms in cell-free systems as well as in macrophages.(ABSTRACT TRUNCATED AT 250 WORDS) Biochem J, 1984 May 1, 219(3), 1009 - 15 Affinity purification and subunit structures of beta-N-acetylhexosaminidases A and B from boar epididymis; Parkes HC et al.; beta-N-Acetylhexosaminidase from boar epididymis was separated into two forms, A and B, on DEAE-cellulose . Both these forms were excluded from Sepharose S-200 and had apparent Mr values of 510 000 on gradient gel electrophoresis under non-denaturing conditions . Affinity chromatography on 2-acetamido-N-(6-aminohexanoyl)-2-deoxy-beta-D-glucopyranosylam ine coupled to CNBr-activated Sepharose 4B was used to separate and purify beta-N-acetylhexosaminidases A and B that had specific activities of 115 and 380 mumol/min per mg of protein respectively . Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of denatured beta-N-acetylhexosaminidase A gave a single major component of Mr 67 000 . beta-N-Acetylhexosaminidase B also had this component, and in addition had polypeptides of Mr 29 000 and 26 000 . All these polypeptides were glycosylated . Antiserum to the B form precipitated form A from solution and reacted with the 67 000-Mr component or form A after electrophoretic transfer from sodium dodecyl sulphate/polyacrylamide gels to nitrocellulose sheets . The 67 000-Mr components of forms A and B yielded identical peptide maps when digested with Staphylococcus aureus V8 proteinase, and the 29 000-Mr and 26 000-Mr components in form B may be related to the 67 000-Mr polypeptide. J Infect, 1984 May, 8(3), 205 - 11 A rabbit model of toxic shock syndrome: clinicopathological features; Arko RJ et al.; The complete pathogenesis of toxic shock syndrome (TSS) has yet to be elucidated . Unmasking the complex interactions among bacterial products, host factors, and possibly tampon components requires a suitable in vivo model . For this purpose, subcutaneous chambers implanted in rabbits were inoculated with Staphylococcus aureus isolated from patients with TSS . Infected rabbits developed illness characterised by multisystem involvement that included periportal inflammation of the liver, erythrophagocytosis in the spleen and lymph nodes as well as extreme vascular dilatation and epithelial lesions similar to those described in patients with TSS . Concentrations of serum creatinine (P less than 0.03) and triglycerides (P less than 0.04) were significantly raised in rabbits infected with TSS strains compared with rabbits infected with non-TSS strains of S . aureus . Both TSS and non-TSS strains of S . aureus produced fever and diarrhoea, but TSS strains were significantly (P less than 0.05) more lethal and more likely to produce respiratory distress and lowered blood pressure . This model may help to prove or disprove proposed mechanisms for the development of TSS. J Bacteriol, 1984 May, 158(2), 689 - 95 Autogenous transduction of phi 11de in Staphylococcus aureus: transfer and genetic properties; Dyer DW et al.; The staphylococcal plasmid phi 11de is capable of transduction in the absence of both a helper bacteriophage and detectable plaque-forming bacteriophage . The mechanism of transfer is distinct from generalized transduction in that it does not transduce chromosomal material and is selective with respect to the plasmid DNA that is transduced . The transductants containing phi 11de have the following characteristics: (i) erythromycin resistance at levels displayed by the donor, (ii) expression of and susceptibility to plasmid incompatibility, (iii) dependence upon the host recombination system during transduction, (iv) complementation of phi 11 mutants, and (v) reactivation of UV-irradiated phage. Infect Immun, 1984 May, 44(2), 434 - 8 In vitro synthesis of the delta-lysin of Staphylococcus aureus; Lee KY et al.; The stability of mRNA for the delta-lysin of Staphylococcus aureus was determined by measuring the residual lysin synthesis after inhibition of DNA-dependent RNA polymerase activity with rifampin . At the late logarithmic-early stationary phase of growth the delta-lysin mRNA was very stable, with a half-life of ca . 20 min . Total cellular RNA was extracted from S . aureus and translated with a modified Escherichia coli S-30 system; delta-lysin was identified amongst the translation products by immunoprecipitation and immunoabsorption . The delta-lysin synthesized in vitro was of a size similar to mature delta-lysin and did not require a signal sequence for secretion from the cell. J Virol Methods, 1984 May, 8(3), 191 - 8 Visualization of herpes simplex virus type 1 attachment to target cells using Staphylococcus aureus as a morphologic tag; Hodnichak CM et al.; Staphylococcus aureus was used as a morphologic tag to allow light microscopic localization of herpes simplex virus type 1 (HSV-1) binding and attachment to HEp-2 target cells . The virus was bound to S . aureus through an anti-HSV-1 linkage . The complex was stable and the attached virus still infectious. J Biol Chem, 1984 Apr 25, 259(8), 5222 - 9 Structural analysis of the hepatic glucagon receptor . Identification of a guanine nucleotide-sensitive hormone-binding region; Iyengar R et al.; {125I-Tyr10}Monoiodoglucagon {( 125I}MIG) was cross-linked to liver membrane glucagon receptors with hydroxysuccinimidyl-p-azidobenzoate, and the products were analyzed by sodium dodecyl sulfate-gel electrophoresis . Autoradiograms of the gel obtained after a 24-h exposure showed one major band at Mr = 63,000 that was sensitive to GTP and excess unlabeled glucagon . Exposure for 7 days showed labeling of an additional Mr = 33,000 species that was also sensitive to excess unlabeled glucagon . The Mr = 33,000 peptide can be obtained by subtilisin, trypsin, elastase, or Staphylococcus aureus V8 protease treatment of the {125I}MIG-occupied receptor in the membrane or in Lubrol-PX solution . In contrast, limited proteolysis of membranes containing vacant receptors results in labeling of a Mr = 24,000 peptide . The Mr = 24,000 peptide specifically binds {125I}MIG in a GTP-sensitive manner . The Mr = 33,000 peptide also retains GTP sensitivity since it releases bound {125I}MIG upon addition of GTP . Elastase treatment of the electroeluted Mr = 33,000 peptide yields the Mr = 24,000 and 15,000 fragments . The Mr = 15,000 peptide is the smallest fragment of the receptor as yet identified . Treatment of the Mr = 63,000 receptor with {125I}MIG cross-linked to it with endo-beta-N-acetylglucosaminidase F results in four distinct fragments with Mr values of 61,000, 56,000, 51,000, and 45,000; prolonged treatment resulted in the accumulation of the last two . Neither the Mr = 33,000 nor the Mr = 24,000 fragment appeared to be substrates for endo-beta-N-acetylglucosaminidase F . These data indicate that glucagon receptor is a glycoprotein of approximately 60,000 daltons which contains at least four N-linked glycans accounting for 18,000 daltons of its mass . Both its glucagon binding function and its capacity to interact with the stimulatory regulator of adenylyl cyclase are contained within a fragment of only approximately 21,000 daltons that does not contain any N-linked glycans . Hormone occupancy of the receptor results in a conformational change so as to expose a region that is susceptible to proteolysis by proteases of varying specificities to yield a peptide of approximately 30,000 daltons that also does not contain N-linked glycans. Am J Obstet Gynecol, 1984 Apr 15, 148(8), 1074 - 9 Variant postpartum toxic shock syndrome with probable intrapartum transmission to the neonate; Chow AW et al.; We report a unique mother-infant pair with variant staphylococcal toxic shock syndrome and probable intrapartum transmission to the neonate . Diagnosis of probable toxic shock was supported by the finding of fever, desquamative skin rash, multi-organ system involvement, and pronounced mucocutaneous manifestations, including strawberry tongue, dermal swelling, pharyngitis, and vulvar edema, although hypotension was absent . Staphylococcus aureus was isolated from the vagina, placenta, chorioamnion, and surface swabs and gastric aspirates of the infant . The isolates produced enterotoxin C but not enterotoxin F, and illness developed in both mother and infant despite preexisting high antibody titers to enterotoxin F and enterotoxin C . This unique mother-infant pair highlights our present lack of knowledge of the precise etiology and pathogenesis of toxic shock syndrome and illustrates the consequent difficulty in clinical diagnosis and laboratory confirmation of this disease in certain patients with atypical presentations. J Mol Biol, 1984 Apr 15, 174(3), 419 - 31 Cleavage of two yolk proteins from a precursor in Caenorhabditis elegans; Sharrock WJ; Four yolk proteins have been identified previously in the nematode Caenorhabditis elegans . However, only two of these proteins ( yp170A and yp170B ) are found among the products of in vitro translation of nematode RNA . The other two yolk proteins ( yp115 and yp88 ) are apparently cleaved from a precursor polypeptide of approximately 180,000 Mr . This precursor has been identified as an in vitro translation product and as a metabolically unstable polypeptide in vivo . It is bound by immunoglobulin G (IgG) specific for yp115 and by IgG specific for yp88 . The immunoadsorbed material yields the same pattern of fragments on partial digestion with Staphylococcus aureus V8 protease regardless of whether anti- yp115 or anti- yp88 IgG is used in the adsorption . Like the yp170 polypeptides, the yp115 / yp88 precursor is synthesized by the intestine and secreted intact . The precursor is evidently cleaved to yield yp115 and yp88 after secretion from the intestine but independent of the presence of the gonad . Thus, cleavage probably occurs in the body cavity of the nematode. Cell Immunol, 1984 Apr 15, 85(1), 191 - 200 Distribution of the major histocompatibility complex antigens on different cellular components of human liver; Lautenschlager I et al.; Monoclonal mouse antibodies to the "framework" determinants of the class I and II molecules of the major histocompatibility complex (MHC) were used to demonstrate the presence of the MHC antigens in human liver . First, the localization of these antigens was demonstrated from frozen section histology with indirect FITC immunofluorescence and the cell component(s) binding the mouse antibody were identified by rabbit marker antisera and indirect TRITC immunofluorescence . Second, the antigen expression on the cell surface was analyzed by the Staphylococcus aureus rosette method from cytological cell smears . All antibodies reacted with cells in the liver sinusoids, both with the Kupffer cells and at least partially with the sinusoidal endothelial cells . The same antisera reacted also with the bile duct cells, though weaker, and with some stromal cells in close proximity of the blood vessels . The vascular endothelial cells of hepatic artery, hepatic vein, and portal vein displayed no reaction . Thus human liver differs strikingly from, e.g., human kidney, where the vascular endothelial cells contain large amounts of MHC antigens on the cell surface . This difference may be one explanation to why liver allografts are less promptly rejected than renal allografts in man. Biochem J, 1984 Apr 15, 219(2), 625 - 34 Complete amino acid sequence of the N-terminal extension of calf skin type III procollagen; Brandt A et al.; The N-terminal extension peptide of type III procollagen, isolated from foetal-calf skin, contains 130 amino acid residues . To determine its amino acid sequence, the peptide was reduced and carboxymethylated or aminoethylated and fragmented with trypsin, Staphylococcus aureus V8 proteinase and bacterial collagenase . Pyroglutamate aminopeptidase was used to deblock the N-terminal collagenase fragment to enable amino acid sequencing . The type III collagen extension peptide is homologous to that of the alpha 1 chain of type I procollagen with respect to a three-domain structure . The N-terminal 79 amino acids, which contain ten of the 12 cysteine residues, form a compact globular domain . The next 39 amino acids are in a collagenase triplet sequence (Gly- Xaa - Yaa )n with a high hydroxyproline content . Finally, another short non-collagenous domain of 12 amino acids ends at the cleavage site for procollagen aminopeptidase, which cleaves a proline-glutamine bond . In contrast with type I procollagen, the type III procollagen extension peptides contain interchain disulphide bridges located at the C-terminus of the triple-helical domain. Biochem J, 1984 Apr 15, 219(2), 425 - 8 Localization of viral-envelope-glycoprotein-binding sites in fibronectin; Julkunen I et al.; Purified viral-envelope glycoproteins from influenza A virus were found to bind to two fragments of the fibronectin molecule . Human plasma fibronectin was digested by leucocyte cathepsin G, and three different fragments, of Mr 30000, 40000 and 12000-140000, with specific binding functions were isolated . Micelles of radiolabelled influenza A glycoprotein were allowed to bind to these fragments immobilized on polystyrene micro-titre wells . The C-terminal 120000-14000-Mr fragments that carry the cell-binding activity bound viral proteins most efficiently, whereas the 40000-Mr gelatin-binding fragment bound considerably less . The N-terminal 30000-Mr Staphylococcus aureus-binding fragment was negative in the assays . Laminin, a basement-membrane protein, also bound viral proteins, though less effectively than fibronectin . The binding was abolished if laminin or fibronectin fragments were pretreated with neuraminidase . This suggests that the sialic acids in the sugar moieties of these glycoproteins are involved in the binding . The affinity of viral-envelope glycoproteins for certain domains of fibronectin and for laminin may play a role in virus-cell interactions. Zh Mikrobiol Epidemiol Immunobiol, 1984 Apr, (4), 93 - 7 {Interrelations in the immune system of Staphylococcus aureus carriers}; Kibort RV et al.; The immune status of 60 S . aureus carriers and 60 donors without carrier state was studied with respect to the interrelations of their antistaphylococcal immunity characteristics and natural resistance factors with the levels of different parameters . In contrast to the donors without carrier state, new interrelations between specific and nonspecific humoral immunity factors were shown to appear in S . aureus carriers, their blood sera having the elevated levels of bactericidal activity and the increased titers of staphylococcal antibodies . The phagocytic activity of the blood in the carriers proved to be below the normal level . The S . aureus carriers were found to have disturbances in the interrelations of their phagocytic activity and specific humoral immunity . The immune status of the carriers as characterized by the interrelations of its parameters is compared with the immune status of normal persons. Am J Vet Res, 1984 Apr, 45(4), 786 - 9 Determination of phagocytosis of 32P-labeled Staphylococcus aureus by bovine polymorphonuclear leukocytes; Dulin AM et al.; A procedure for the measurement of phagocytosis by bovine polymorphonuclear leukocytes (PMN) of 32P-labeled Staphylococcus aureus was modified so that a larger number of samples could be compared in a single run, and smaller volumes of sample, PMN, and 32P-labeled S aureus could be used . Results were highly reproducible, with a coefficient of variation between duplicate determinations of less than or equal to 2% . Lysostaphin was prepared from the supernatant of S staphylolyticus and was compared with a commercially available preparation . Effects of lysostaphin on PMN and influence of incubation media on release of 32P from 32P-labeled S aureus by lysostaphin were examined. Monatsschr Kinderheilkd, 1984 Apr, 132(4), 217 - 21 {Toxic shock syndrome in a 14-year-old girl}; Luders D et al.; The toxic shock syndrome in a 14-year old girl is described . This syndrome occurs most frequently - but not exclusively - in the teens and young women during the first days of menstruation, if tampons are used . The patients are acutely ill with high fever, diarrhea and/or vomiting, with a rash, with loss of consciousness, and signs of shock (occasionally shock lung syndrome and renal insufficiency) . During convalescence desquamation of hands and feet shows up . Patients with much less severe symptoms have been seen . The primary lesion is a local infection (e.g . vaginitis) with staphylococcus aureus, the symptoms being caused by staphylococcal toxins . Early recognition and immediate therapy are important for a better prognosis . The therapy consists of removal of the tampon, i.v . fluids including albumin, and the administration of a beta-lactamase-resistant antibiotic. J Dairy Sci, 1984 Apr, 67(4), 826 - 34 Neutrophil migration through test end tissues of bovine mammary quarters experimentally challenged with Staphylococcus aureus; Nickerson SC et al.; Diapedesis and infiltration of neutrophils into internal epithelial tissues of the distal teat end and migration into milk were studied in bovine mammary quarters infected with Staphylococcus aureus . Neutrophil extravasation, penetration of the epithelium, and mode of passage into milk were evaluated by light and transmission electron microscopy . Collection of observations from 10 infected quarters of six cows provided morphologic evidence for the following sequence of events as neutrophils passed from blood into milk . In capillaries of the subepithelial stroma, neutrophils adhered to luminal walls, penetrated endothelia and basal lamina, then migrated across the periendothelial cell layer into extravascular connective tissues adjacent to epithelial linings . The leukocytes then penetrated epithelial basal laminae and migrated between basal epithelial cells to gain access to the luminal cell layer . Possible modes of migration across luminal cells into milk included 1) projection through individual, degenerate luminal cells, 2) penetration between intact epithelia, and 3) passage into milk as luminal cells desquamated in areas of epithelial metaplasia . The first method appeared to be the predominant mechanism of migration . These data suggest that elevated numbers of neutrophils in distal teat end epithelium and in cisternal milk may be instrumental in the initial events that prevent establishment of infection in the bovine mammary gland. J Antibiot (Tokyo), 1984 Apr, 37(4), 325 - 9 A new naphthaquinone antibiotic from a new species of yeast; Flegel TW et al.; A fusiform yeast producing limited pseudomycelium and limited true mycelium on malt extract agar has been isolated . This non-fermentative yeast has hyaline cell walls but produces a thick, black oily exudate which is water insoluble and gives the colony a smooth black lacquered appearance . On the basis of morphology and physiology, this organism is distinctive enough to warrant the designation of a new genus . During stationary phase of cultures on chemically defined medium, a deep red substance is produced which has strong antibiotic activity against Staphylococcus aureus in vitro . The substance has been identified as a new natural naphthaquinone of the empirical formula C16H16O7 . Eur J Clin Microbiol, 1984 Apr, 3(2), 108 - 12 Opsonic activity of fibronectin in the phagocytosis of Staphylococcus aureus by polymorphonuclear leukocytes; Eriksen HO et al.; The effect of the opsonic activity of human purified fibronectin on phagocytosis of Staphylococcus aureus by human polymorphonuclear leukocytes was investigated . After opsonization with fibronectin there was a significant increase in the rate of phagocytosis of four out of six Staphylococcus aureus strains . Both attachment to and ingestion by leukocytes was affected, as revealed by lysostaphin treatment . Incubation of leukocytes with fibronectin prior to phagocytosis did not enhance the phagocytosis of Staphylococcus aureus . This shows that the observed enhancement of phagocytosis was dependent on the binding of fibronectin to Staphylococcus aureus . The ability of fibronectin to opsonize different strains of Staphylococcus aureus varied with the strain . A variation was also observed in fibronectin-induced phagocytosis by leukocytes from different donors. Can J Comp Med, 1984 Apr, 48(2), 125 - 9 Fatal mastitis of dairy cows: a retrospective study; Hazlett MJ et al.; The necropsy records of dairy cows with mastitis were reviewed from the provincial veterinary laboratory in Guelph (44 cases of mastitis in nine years) and from the Ontario Veterinary College (168 cases in 14 years) . Mastitis was considered to be the primary cause of death in 167 of 212 cows (79%) . Of these 167 cases of mastitis, Escherichia coli was involved in 107 (64%), Klebsiella sp . in 12 (7%) and Staphylococcus aureus in 11 (7%) . Bacteriology was not reported in 22 cases . Coliform mastitis, the most commonly identified type of fatal mastitis, was characterized histologically by the presence of infarcted areas in affected glands and by the lack of demonstrable bacteria, and was thus easily identified from fatal mastitis caused by S . aureus. Biol Reprod, 1984 Apr, 30(3), 775 - 86 Production and characterization of monoclonal antibodies to the sperm acrosome stabilizing factor (ASF): utilization for purification and molecular analysis of ASF; Reynolds AB et al.; Utilizing hybridoma technology and highly purified acrosome stabilizing factor (ASF), six monoclonal antibodies (mAbs) specific for ASF were produced and characterized . Specificity and binding properties of each clone were examined by immunoperoxidase labeling of electrophoretic blots of rabbit serum, seminal plasma, cauda epididymal fluid and vasectomized seminal plasma separated on native and sodium dodecyl sulfate (SDS)-polyacrylamide gels . All mAbs recognize ASF in seminal plasma and cauda epididymal fluid but do not bind components in serum or vasectomized seminal plasma . Purification of ASF by affinity chromatography utilizing the mAbs, has shortened the 6-day isolation procedure for ASF used previously to less than 2 h and has increased the yield from 2 micrograms to 300 micrograms of ASF obtained per ml of seminal plasma . Three mAbs were used in conjunction with Cleveland digest with Staphylococcus aureus V8 protease, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoperoxidase labeling of Western Blots, to identify peptides containing specific determinants recognized by each mAb . At least five separate determinants were recognized by the six mAbs . The sensitivity of the Western blotting technique in conjunction with the specificity of the mAbs was exploited to detect polymeric forms of ASF in seminal plasma and cauda epididymal fluid . ASF is shown to be a 360-kd dimer consisting of two identical 180-kd monomers . Tools are now available to develop sensitive qualitative and quantitative assays for ASF, thus providing rapid, extremely sensitive methods for evaluating experiments designed to probe the molecular mechanism of capacitation. J Clin Microbiol, 1984 Apr, 19(4), 552 - 4 Detection of antibody to staphylococcal lipoteichoic acid with a microenzyme-linked immunosorbent assay; Carruthers MM et al.; Sera from individuals with Staphylococcus aureus endocarditis and osteomyelitis and from some individuals with other forms of gram-positive endocarditis yielded higher readings in a microenzyme-linked immunosorbent assay against lipoteichoic acid from S . aureus than did sera from individuals with other types of serious staphylococcal infection or non-staphylococcal osteomyelitis, or from unselected inpatients. J Clin Microbiol, 1984 Apr, 19(4), 511 - 5 Comparison of a new enzyme-linked immunosorbent assay method with counterimmunoelectrophoresis for detection of teichoic acid antibodies in sera from patients with Staphylococcus aureus infections; Herzog C et al.; Ribitol-teichoic acid antibodies were measured by a new enzyme-linked immunosorbent assay (ELISA) and by counterimmunoelectrophoresis in serum samples from 47 patients with serious Staphylococcus aureus infections, 63 infected patients, and 177 healthy controls . The same antigen was used for both tests . The group of patients with S . aureus endocarditis (6 patients) had significantly higher ELISA readings than the patients with other deep-seated infections (26 patients) or with an uncomplicated S . aureus bacteremia (15 patients) . The patients with other serious gram-positive (40 patients) or gram-negative (23 patients) infections did not differ from the healthy control group . There were only three (7.5%) low-level cross-reactions among the infections caused by gram-positive organisms other than S . aureus . Of 46 initially ribitol-teichoic acid antibody-negative patients followed up for 2 weeks or more, only those developing a serious S . aureus infection showed a significant rise of the ELISA reading . There was a good correlation between ELISA and counterimmunoelectrophoresis . Both tests could be useful in the diagnosis and the management of complicated S . aureus infections . The ELISA method is, however, more sensitive and usually reflects the antibody rise after an infection earlier than does counterimmunoelectrophoresis. Ann Surg, 1984 Apr, 199(4), 438 - 44 The walk-in anergic patient . How best to assess the risk of sepsis following elective surgery; Christou NV et al.; This prospective study evaluated host resistance in a surgical population who walked into the hospital for elective surgery . Patients were stratified into Hospital Reactive (HR, n = 19) if they reacted to two or more of five recall skin test antigens and Walk-in Anergic (WA, n = 26) if they did not react to the antigens . The WA patients were slightly older (74.4 +/- 1.8 years, +/- SEM versus 66.7 +/- 2.7 p less than 0.05) . Diagnosis in the HR and WA group were: tumors 13/19 versus 21/26, diverticulitis 3/19 versus 0/19, and miscellaneous 3/19 versus 5/26 . Twenty-five laboratory normal controls (LN) were also studied . There were no significant differences in the following parameters between the HR and WA groups: stage of disease; hemoglobin; circulating leukocyte count; polymorphonuclear cell counts; total lymphocyte counts (both groups lower than LN, p less than 0.05), monocyte counts (both higher than LN, p less than 0.05); per cent E-rosettes and lymphocyte blastogenesis to mitogens (phytohemagglutinin, concanavalin-A) and antigens (purified protein derivative and tetanus); phagocytosis of preopsonised Staphylococcus aureus 502A, at 5, 10, and 20 minutes; alpha, beta, and gamma globulins; C3, and total hemolytic complement (CH50) levels; C-reactive protein; and ANA and DNA levels . The HR group demonstrated an increase in the rate of killing of Staphylococcus 502A at 10, 20, 40, and 80 minutes compared to the LN group but the WA group did not show this augmentation (p less than 0.001) . The serum albumins were: LN = 4.46, HR = 3.98, WA = 3.43 g/dl (p less than 0.05) . Degree and duration of surgery was the same in the HR and WA groups . There were no major sepsis episodes (bacteremia or proven intracavitary abscess) in the HR patients versus 25% in the WA patients (p less than 0.05) . There was one death (6%, pulmonary embolus) in the HR group and 8 (40%) in the WA group (p less than 0.05) . Antibiotic prophylaxis was equal but the WA patients received therapeutic antibiotics more frequently (65% versus 11% p less than 0.05) . Of all the host immunocompetence tests measured in this study, the delayed type hypersensitivity skin test response and the serum albumin were variables abnormal between the survivors and those who died. Surg Gynecol Obstet, 1984 Apr, 158(4), 349 - 53 Penetration of clindamycin into experimental Staphylococcus aureus infections; Simon GL et al.; Inactivation of clindamycin at the site of experimental infection with Staphylococcus aureus was studied using rabbits with plastic capsules implanted in the peritoneal cavity . The mean percentage penetration of bioactive clindamycin (concentration in capsule divided by simultaneous concentration in serum times 100) into infected and noninfected capsules was 30.4 per cent and 13.0 per cent, respectively . In contrast, the mean penetration of radiolabeled clindamycin into infected capsules was 38.4 per cent . These findings indicate that the observed loss of bioactivity in infected capsules is due to intracapsular inactivation of clindamycin and not to an alteration in capsular permeability . Biologic inactivation of clindamycin was not evident after in vitro incubation of the drug with Staphylococcus aureus . These results suggest that the observed loss of bioactivity may be due to chemical modification by enzymes in the inflammatory exudate or to binding of the antibiotic to tissue components. Antimicrob Agents Chemother, 1984 Apr, 25(4), 491 - 3 Subinhibitory concentrations of imipenem induce increased resistance to methicillin and imipenem in vitro in methicillin-resistant Staphylococcus aureus; Forbes BA et al.; Methicillin-resistant (MR) Staphylococcus aureus that was susceptible to less than 0.75 micrograms of imipenem per ml demonstrated inducible resistance . MR S . aureus preincubated with 0.05 microgram of imipenem per ml grew in medium with an imipenem concentration of 32 micrograms/ml, and methicillin MICs increased 20-fold . Non-MR S . aureus exhibited no induction . Preincubation with methicillin produced no effect . Induction appeared to be a unique interaction of imipenem with MR S . aureus. Proc Natl Acad Sci U S A, 1984 Apr, 81(8), 2475 - 8 Development of a human T-cell hybridoma secreting separate B-cell growth and differentiation factors; Butler JL et al.; A cloned human T-cell hybridoma (7D5) secreting B-cell growth factor (BCGF) and B-cell differentiation factor (BCDF) was established . Supernatant from this hybrid was capable of maintaining proliferation in anti-IgM-activated normal human B cells . In addition, the hybridoma supernatant induced differentiation and antibody secretion in Staphylococcus aureus Cowan I-stimulated B cells . No interleukin 2 was present in supernatant from this hybridoma . Molecular size of the hybridoma-derived BCGF and BCDF was determined by gel filtration chromatography . BCGF activity was present in the 20-kDa fractions, and BCDF activity eluted in the 30- to 35-kDa fractions . The isoelectric points of the factors, determined by chromatofocusing, were 6.6 for BCGF and 5.9 for BCDF . Finally, absorption experiments were performed using specific target cells . Phytohemagglutinin-stimulated T-cell blasts did not remove either BCGF or BCDF activity . Anti-IgM-activated B cells absorbed BCGF but not BCDF . In contrast, CESS cells removed BCDF but not BCGF . Thus, a human T-cell hybridoma secreting two distinct B-cell lymphokines was developed . Further immunochemical and functional studies of these immunoregulatory molecules should greatly enhance our understanding of the regulation of human B-cell function in normal and disease states. Arch Otolaryngol, 1984 Apr, 110(4), 228 - 31 beta-Lactamase-producing bacteria recovered after clinical failures with various penicillin therapy; Brook I; The presence of beta-lactamase-producing bacteria in clinical specimens was investigated in 185 children with orofacial or respiratory tract infections . All of these patients failed to respond to antimicrobial therapy, including penicillins, that was administered to 148 (80%) of them . beta-Lactamase-producing aerobic and anaerobic bacteria were detected in 75 (40.5%) of the 185 children . The beta-lactamase-producing strains included all 11 strains of the Bacteroides fragilis group, 30 (45.4%) of the 66 strains of Bacteroides melaninogenicus group, five (41.7%) of the 12 strains of Bacteroides oralis, and 41 (97.6%) of 42 strains of Staphylococcus aureus . All beta-lactamase-producing Bacteroides strains were resistant to penicillin as compared with the non-beta-lactamase-producing strains . Clinical cure was achieved after surgical drainage and a change in antimicrobial therapy in most of the patients . In treatment of orofacial and respiratory tract infections, the clinician should consider the presence of beta-lactamase-producing Bacteroides sp and S aureus as a possible cause of clinical failure with various penicillin therapies. J Immunol, 1984 Apr, 132(4), 1858 - 62 Human B cell-inducing factor(s) for production of IgM, IgG and IgA: independence from IL 2; Ralph P et al.; Human peripheral blood B cells are stimulated into proliferation by killed Staphylococcus aureus bacteria strain Cowan I (Sac) . T lymphocytes in the presence of a T cell mitogen induce high numbers of immunoglobulin-secreting cells (ISC) in these Sac-stimulated B cells . The T cells can be largely replaced by a lymphokine factor . We describe here the 11000-fold purification of this B cell-inducing factor (BIF) . BIF preparations that are free of IL 2 do not require IL 2 for optimal induction of ISC . This was shown by the lack of effect of IL 2 alone or with suboptimal or optimal concentrations of BIF on the induction of ISC and by the absence of IL 2 production in the purified B cell population which, with other controls, excludes significant T cell contamination . BIF, purified through four fractionation steps and free of IL 2, induces IgM, IgG, and IgA-ISC in approximately the same ratio as unfractionated lymphokine . Because we have not yet attained a pure BIF preparation, the possibility of separate factors for the production of each immunoglobulin isotype cannot be ruled out. J Comp Pathol, 1984 Apr, 94(2), 183 - 96 Histopathology and pathogenesis of mouse mastitis induced with Staphylococcus aureus mutants; Haraldsson I et al.; Mammary glands of mice were inoculated with a well-defined strain of Staphylococcus aureus or with derived mutants lacking alpha-haemolysin, coagulase or protein A, in order to evaluate the pathogenicity of these factors . Alpha-haemolysin-negative and coagulase-negative mutants showed less virulence than the wild-type strain . Various protein A-negative mutants gave contradictory results . The lesions ranged from consistently non-reactive necrosis of the entire mammary gland to a limited inflammatory reaction . The necroses, especially when affecting the whole mammary gland, were mostly associated with vascular lesions . Because the necroses were non-reactive, the vascular lesions were considered to be primary, but they could not be linked exclusively with the effect of alpha-haemolysin and a multifactorial aetiology seemed most probable . After inoculation with coagulase-lacking bacteria the development of parenchymal lesions was delayed, hypothetically because of increased phagocytic activity . The investigation indicated that protein A could have influenced the pathogenesis of the lesions, since all but one of the mutants lacking protein A showed low virulence, whereas a high protein A-producing, haemolysin-negative mutant was as virulent as the wild-type strain . A connection between protein A, bacterial adherence and bacterial growth rate is therefore conceivable . The possibility cannot be excluded, however, that during mutagenesis some as yet unknown cell surface factors were affected. J Clin Microbiol, 1984 Apr, 19(4), 464 - 7 Evaluation of the automicrobic system for detection of resistance of Staphylococcus aureus to methicillin; Woolfrey BF et al.; The AutoMicrobic system (AMS) (Vitek System, Inc., Hazelwood, Mo.) was tested for its ability to determine oxacillin and gentamicin susceptibility of 98 known oxacillin-susceptible and 103 known oxacillin-resistant Staphylococcus aureus isolates . AMS and reference oxacillin susceptibility results were in agreement for all 95 (100%) oxacillin-susceptible isolates . In contrast, only 23 (22.3%) of the 103 known oxacillin-resistant isolates were correctly reported . For the known oxacillin-resistant isolates, 65 received AMS reports at 3 to 4 h, with only 9% being correct, whereas 38 were reported at 5 to 6 h, with 47% being correct . The reliability of AMS gentamicin susceptibility results was evaluated by testing the 198 S . aureus isolates in parallel with MIC-2000 broth dilution tests . AMS gentamicin susceptibility results were found to be reliable and essentially identical to MIC-2000 results . The possibility of improving AMS oxacillin resistance detection by using gentamicin resistance as a linked screening marker for oxacillin resistance was evaluated with data from the parallel AMS and MIC-2000 gentamicin susceptibility tests and from data accrued on recent clinical laboratory isolates . By these two approaches, respective sensitivities of 97 and 99.8%, and specificity of 72%, were found for detection of oxacillin-resistant isolates by using gentamicin resistance as a marker. J Urol, 1984 Apr, 131(4), 818 - 21 Absorption of doxorubicin-hydrochloride and mitomycin-C after instillation into noninfected and infected bladders of dogs; Schmidbauer CP et al.; The described investigations were carried out in order to determine the degree of absorption of doxorubicin, and mitomycin-C after intravesical instillation into noninfected or Staphylococcus aureus-infected bladders in beagle dogs . The drug concentrations in the bladder wall were determined using diffusion chambers with permeable membranes . Two hours after end of instillation of 10 mg . doxorubicin, a concentration of 1.4 ng . per ml . was measured in the bladder wall of noninfected animals, and 3.75 ng . per ml . in that of infected animals (p less than 0.05) . The simultaneously measured serum concentration reached mean peak levels after 30 minutes . The concentrations in infected animals were 3 times higher (1.9 ng . per ml.) than in noninfected animals (0.6 ng . per ml.) (p less than 0.05) . After instillation of 1 mg . per kg . bw . mitomycin-C the concentration in both groups of animals was below 0.06 micrograms per ml . Doxorubicin concentrations were determined with a radioimmunoassay and mitomycin-C with a micro-agar diffusion method. Can J Microbiol, 1984 Apr, 30(4), 470 - 4 Evaluation of a pour-plate system with a rabbit plasma-bovine fibrinogen agar for the enumeration of Staphylococcus aureus in food; Beckers HJ et al.; Investigations were carried out concerning the selectivity and productivity of rabbit plasma fibrinogen (RPF) agar according to Beckers et al . (H . J . Beckers, F . M . van Leusden, W . M . Hogeboom, and E . H . M . Delfgou-van Asch . 1980 . De Ware(n)-Chemicus, 10: 125-130) . Its selectivity was compared with pork plasma fibrinogen (PPF) medium according to Hauschild et al . (A . H . W . Hauschild, C . E . Park, and R . Hilsheimer . 1979 . Can . J . Microbiol . 25: 1052-1057) and its productivity was compared with PPF medium and Baird-Parker's egg yolk tellurite glycine pyruvate (ETGP) agar . In total 139 samples of naturally contaminated foodstuffs were examined . RPF agar scored higher than ETGP agar; although only small (mean value of differences 0.09 log units), the differences were statistically significant . While no significant differences in sensitivity between RPF agar and PPF medium were encountered, RPF agar was statistically more selective than PPF medium . It is concluded that RPF agar is very suitable for the enumeration of Staphyloccus aureus in foods. J Gen Microbiol, 1984 Apr, 130 ( Pt 4), 907 - 17 Surface antigens of intact Aspergillus fumigatus mycelium: their localization using radiolabelled protein A as marker; Hearn VM; Isotopically-labelled Protein A from Staphylococcus aureus was used as a marker to quantify binding of IgG molecules to surface antigens of Aspergillus fumigatus mycelium . The IgG class antibodies were obtained from rabbits inoculated with mycelial fractions and from patients suffering from aspergillosis and aspergillus-related diseases . The highest incorporation of label was obtained with an antiserum from rabbits inoculated with A . fumigatus wall material . Antibodies raised to other antigenic fractions of A . fumigatus and antibodies from patients infected with aspergillus gave lower levels of incorporation of Protein A . In competitive binding experiments, pre-incubation of antibodies with partially purified aspergillus antigens depressed subsequent binding to the mycelial surface by 20-30% of that of the control values . Low molecular weight disaccharides and oligosaccharides were without effect in this system . Preincubation of A . fumigatus with lectins having specificities for defined sugar residues did not reduce subsequent antigen/antibody interaction . Treatment of the mycelial surface with certain proteolytic or polysaccharolytic enzymes led to a decrease in antibody binding, while pretreatment of A . fumigatus with the hydrolytic enzyme mixture of Trichoderma harzianum culture filtrate gave increased antibody binding . Aspergillus species showed different susceptibilities to enzyme action and their surface structures could be differentiated from A . fumigatus on this basis . These differences were not obvious in direct binding experiments where antibodies raised to A . fumigatus wall bound with equal facility to antigenic sites located on the walls of other Aspergillus species. Acta Pathol Microbiol Immunol Scand {C}, 1984 Apr, 92(2), 121 - 8 Induction of immunoglobulin secretion in cultured human lymphocytes by 4 Staphylococcus aureus strains and their extracts; Effersoe H et al.; Human blood lymphocytes were stimulated in vitro by four Staphylococcus aureus strains . Activation of immunoglobulin-secreting cells was determined by a reverse plaque forming cell (PFC) assay, and proliferation by quantitation of thymidine incorporation . Whole killed S . aureus were slightly more efficient than water-soluble preparations in the form of sonicated extracts and culture supernatants . Two S . aureus strains rich in protein A (Cowan I and E 2371) and one S . aureus strain deficient in protein A (E 1369) were potent B-lymphocyte stimulators inducing maximal activity on day 6 of culture . Another S . aureus strain deficient in protein A (Wood 46) did not possess++ human lymphocyte stimulating capacity. J Appl Bacteriol, 1984 Apr, 56(2), 215 - 20 A simplified system for biotyping Staphylococcus aureus strains isolated from animal species; Devriese LA; A biotyping system for Staphylococcus aureus strains is proposed which is a simplified version of biotyping procedures described in the literature . It differentiates Staph . aureus strains from man and animals into host-specific ecovars and biotypes which are not host-specific . With the help of tests for beta-haemolysin, staphylokinase , coagulation of bovine plasma and the crystal-violet reaction, the origin of many but not all Staph . aureus strains can be determined: 604 of 809 strains from man, poultry, cattle, pigs, goats, rabbits and foods could be alloted to four ecovars which are typically associated with man, poultry, sheep and goats and cattle . The other strains belonged to five non-host specific biotypes. Acta Endocrinol (Copenh), 1984 Apr, 105(4), 492 - 9 Simple and sensitive method for estimation of antithyroid plasma membrane antibodies in the serum of patients with autoimmune thyroid diseases; comparison with other assays; Gardas A et al.; The ability of protein A from Staphylococcus aureus to interact with Fc fragments of IgG was used to estimate the antithyroid plasma membrane antibodies in sera of patients with Graves' disease . The results were expressed as an antithyroid plasma membrane antibodies (ATMA) index . The ATMA index estimated in 60 healthy blood donors varied from 0.57 to 1.28, with a mean value of 0.99, SD theta 0.20 . The ATMA index in hyperthyroid untreated Graves' disease varied from 1.80 to 8.0, with a mean value of 4.7 . Autoantibody binding to thyroid plasma membranes could be inhibited by (Fab)2 fragments obtained from the serum of patients with Graves' disease but not by (Fab)2 fragments obtained from the serum of healthy blood donors . The influence of rabbit antithyroglobulin and antimicrosomal antibodies on the ATMA index estimation has been evaluated . The ATMA index estimation was compared with the thyrotrophin binding inhibiting immunoglobulins (TBII) index and with the adenyl cyclase stimulating activity of immunoglobulins obtained from 92 hyperthyroid Graves' patients . The ATMA index was positive in 97%, the TBII index in 62% and TSI in 35% of cases . This method using protein A could also be used for estimation of ATMA in other autoimmune thyroid disorders. Proc Natl Acad Sci U S A, 1984 Apr, 81(8), 2553 - 7 Binding of alpha-bungarotoxin to proteolytic fragments of the alpha subunit of Torpedo acetylcholine receptor analyzed by protein transfer on positively charged membrane filters; Wilson PT et al.; Proteolytic fragments of the alpha subunit of the acetylcholine receptor retain the ability to bind alpha-bungarotoxin following resolution by polyacrylamide gel electrophoresis and immobilization on protein transfers . The alpha subunit of the acetylcholine receptor of Torpedo electric organ was digested with four proteases: Staphylococcus aureus V-8 protease, papain, bromelain, and proteinase K . The proteolytic fragments resolved on 15% polyacrylamide gels were electrophoretically transferred onto positively charged nylon membrane filters . When incubated with 0.3 nM 125I-labeled alpha-bungarotoxin and autoradiographed, the transfers yielded patterns of labeled bands characteristic for each protease . The molecular masses of the fragments binding toxin ranged from 7 to 34 kDa, with major groupings in the 8-, 18-, and 28-kDa ranges . The apparent affinity of the fragments for alpha-bungarotoxin as determined from the IC50 value was 6.7 X 10(-8) M . The labeling of fragments with alpha-bungarotoxin could be inhibited by prior affinity alkylation of receptor-containing membranes with 4-(N-maleimido)-alpha-benzyltrimethylammonium iodide . These findings demonstrate that immobilized proteolytic fragments as small as 1/5 the size of the alpha subunit retain the structural characteristics necessary for binding alpha-bungarotoxin, although the toxin is bound to the fragments with lower affinity than to the native receptor . The effect of affinity ligand alkylation demonstrates that the alpha-bungarotoxin binding site detected on the proteolytic fragments is the same as the affinity-labeled acetylcholine binding site on the intact acetylcholine receptor. Mol Cell Biol, 1984 Apr, 4(4), 688 - 94 Fatty acid-acylated proteins in secretory mutants of Saccharomyces cerevisiae; Wen D et al.; Yeast secretory (sec) mutants that are blocked in the transport of secretory proteins and accumulate membrane organelles were used to study the biosynthesis of fatty acid-acylated proteins . Four proteins were labeled with {3H}palmitate in sec mutants accumulating endoplasmic reticulum membranes . Three of these (molecular weights approximately equal to 20,000, 50,000, and 120,000) were N-linked glycoproteins, based on their ability to be labeled with {3H}mannose and their sensitivity to endoglycosidase H . The fourth protein (molecular weight approximately equal to 30,000) also was labeled with {3H}mannose but was insensitive to endoglycosidase H; it appeared to contain O-linked sugars . In sec mutants accumulating Golgi membranes or post-Golgi vesicles, a 35-kilodalton protein was labeled with {3H}palmitate . Analysis of Staphylococcus aureus protease V8 digests and pulse-chase experiments indicated that the 30-kilodalton protein was a precursor of 35 kilodaltons . None of these proteins was labeled with {3H}palmitate in a sec mutant that blocked the penetration of nascent polypeptides into endoplasmic reticulum; thus, acylation occurred in endoplasmic reticulum . All four proteins could be recovered from fractions enriched for yeast membranes . Fatty acids were not released from proteins by boiling in sodium dodecyl sulfate or extraction with organic solvents but were recovered as methyl esters after proteins were treated with KOH-methanol, a reaction characteristic of an acyl ester linkage. J Cell Biol, 1984 Apr, 98(4), 1497 - 504 A domain-specific marker for the hepatocyte plasma membrane . III . Isolation of bile canalicular membrane by immunoadsorption; Roman LM et al.; Previous immunolabeling studies (Roman, L.M., and A.L . Hubbard, 1983, J . Cell Biol., 96:1548-1558; Roman, L.M., and A.L . Hubbard, 1984, J . Cell Biol., 98:1488-1496, companion paper) established leucine aminopeptidase (LAP) as a specific marker for the bile canalicular (BC) domain of the rat hepatocyte plasma membrane (PM) . In this study, we have isolated membrane from a sonicated PM vesicle fraction using anti-LAP-coated Staphylococcus aureus cells as a solid-phase immunoadsorbent . The extent and specificity of the immunoadsorption were assessed by following the behavior of LAP (the BC marker) and 32P-labeled membrane phospholipids (a uniform membrane marker) . The BC fraction obtained was significantly enriched in LAP (yield: greater than 70% of PM-LAP) . Alkaline phosphatase, 5'-nucleotidase, and a 110,000-dalton glycoprotein, HA-4, were enriched in the BC fraction to the same extent as LAP (enzyme or antigen/LAP = 1.0) . However, alkaline phosphodiesterase I was not enriched to the same degree (enzyme/LAP = 0.5) . Contamination of this BC fraction by membrane derived from the sinusoidal domain and endoplasmic reticulum, as determined from the distribution of the asialoglycoprotein receptor and NADH cytochrome c reductase, respectively, was small (less than 13%). Br J Anaesth, 1984 Apr, 56(4), 333 - 8 Impaired B lymphocyte function during open-heart surgery . Effects of anaesthesia and surgery; Eskola J et al.; B lymphocyte function in vitro was measured in patients undergoing open-heart surgery . Conventional balanced anaesthesia, or high-dose fentanyl anaesthesia was used . Pokeweed mitogen (PWM) induced lymphocyte transformation was depressed at the end of the operation, but the response to formalinized Staphylococcus aureus Cowan I (StaCw) was not . The numbers of immunoglobulin producing and secreting cells measured by an indirect protein A plaque-forming cell assay decreased after PWM-stimulation, but remained unchanged after StaCw stimulation at the end of the operation . IgG, IgM and IgA secretion by PWM- and StaCw-stimulated lymphocytes, into the culture medium, was depressed in the period after operation . Depressed immune functions occurred after open-heart surgery, but not in association with anaesthesia alone before surgery . The decreases were not mediated by hydrocortisone-sensitive suppressor cells . Minor differences between the two anaesthetic techniques were found in lymphocyte proliferative responses. J Trauma, 1984 Apr, 24(4), 323 - 6 A model of experimental post-traumatic osteomyelitis in guinea pigs; Passl R et al.; A model of experimental post-traumatic osteomyelitis is described in which the femur of guinea pigs was fractured and infected with E . coli (10(5)) or Staphylococcus aureus (10(4)) . Traumatized uninfected animals served as controls . The animals were further divided within each group by treating the fractured site with an intramedullary wire in one half . Osteomyelitis developed and became chronic in all guinea pigs infected with Staph . aureus, and in nine of 12 infected with E . coli . All animals infected with E . coli treated with an intramedullary wire developed chronic osteomyelitis; only four of seven from E . coli-infected animals with fractures developed this disease . Moreover, Staph . aureus could be recovered from the osseous tissue in the chronic stage of the disease regularly, while E . coli was only present in the early weeks after operation, but not in the chronic stage. Clin Orthop, 1984 Apr, (184), 114 - 7 Efficacy of a topical antibiotic irrigant in decreasing or eliminating bacterial contamination in surgical wounds; Benjamin JB et al.; Using a simple in vitro system, the authors showed that colony counts of Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas species can be reduced by 12%-56% with saline irrigation; the reduction of colony numbers, however, especially for S . aureus, was not always statistically significant . However, even if it were statistically significant, the amount of reduction would not be clinically significant . A topical antibiotic irrigant containing bacitracin/neomycin was effective against S . aureus, S . epidermidis-, and E . coli-treated agar plates . Except for a single plate containing Pseudomonas organisms, the growth of Pseudomonas colonies was also prevented by antibiotic irrigation . Tissue samples of muscle, fat, and bone obtained during operation showed antibiotic levels comparable with, and in most cases greater than, those found in the blood agar plates . These data may be clinically significant and suggest that bacitracin/neomycin irrigation can be safely used to reduce the incidence of postoperative infection. Can J Microbiol, 1984 Apr, 30(4), 461 - 9 In vivo proteins of defective interfering particles of poliovirus; Tershak DR; DX particles of poliovirus are deletion mutants that do not induce synthesis of capsid proteins or the precursor of capsid proteins (NCVPla) during infection . However, cells infected with DX particles synthesize two proteins, p68 and p25, that are not detected during growth of standard virus, and a protein of 27 000 (p27) which is comparable in molecular weight to VP3 . Peptide maps of these proteins were obtained by partial digestion with Staphylococcus aureus V8 protease and elastase . The peptide map of p68 corresponded approximately 70% with the peptide map of NCVPla, and antiserum against virions reacted with p68 . These data suggest that p68 is a large fragment of NCVPla . Digestion of purified structural proteins VP1, VP2, and VP3 yielded distinct peptide maps, but p25 was resistant to both V8 protease and elastase and did not react noticeably with anticapsid antibody . Peptide maps obtained for in vivo viral proteins migrating with a molecular weight of 27 000 were complex, indicating the presence of at least two and possibly three proteins . Cells infected with standard gs and gr viruses produced authentic VP3, but cells infected with defective interfering particles did not . However, one gr variant of standard virus contained a mutation in structural protein VP2. Acta Pathol Microbiol Immunol Scand {B}, 1984 Apr, 92(2), 115 - 8 Recognition of coagulase-negative Staphylococcus aureus strains by primary polymyxin susceptibility testing; Heltberg O et al.; A prospective study evaluating the usefulness of polymyxin susceptibility testing in unveiling coagulase-negative S . aureus strains was undertaken . During a six months study period, 14 staphylococcal isolates from four patients were initially found to lack coagulase activity and to be polymyxin resistant; in comparison, approximately 1500 ordinary, coagulase-positive, polymyxin resistant S . aureus isolates were found during the same period . One isolate from each of the four patients together with a previously isolated coagulase-negative S . aureus strain from our own collection were further characterized . Two of the strains turned out to show delayed coagulase activity on retesting, while the other three strains failed to produce clotting in any coagulase assay . Judged by thermostable nuclease activity, phage typing and biochemical profiles these three coagulase-negative strains are bona fide S . aureus strains . The polymyxin test thus appears to be useful in disclosing S . aureus variants not identified as S . aureus by routinely performed coagulase assays. Thromb Res, 1984 Apr 1, 34(1), 35 - 49 Comparison of platelet fibrinogen receptors on intact and proteolytically-treated platelets by use of an anti-glycoprotein IIIa monoclonal antibody (MA 123); Kornecki E et al.; A murine monoclonal antibody (MA 123) was selected by screening 153 supernatants of hybridoma cells secreting anti-human platelet antibodies for their ability to inhibit the fibrinogen-induced aggregation of chymotrypsin-treated platelets . MA 123 inhibited the binding of 125I-fibrinogen to ADP-stimulated intact human platelets and to platelets treated with chymotrypsin or pronase . Moreover, it inhibited the fibrinogen-induced aggregation of these platelet suspensions . The degree of inhibition was similar in each of the three types of platelets tested . The interactions of MA 123 with the 125I-labeled surface components of intact and chymotrypsin-treated platelets were studied by immunoprecipitation using Staphylococcus aureus coated with goat anti-mouse IgG, followed by SDS-polyacrylamide gel electrophoresis and autoradiography . MA 123 precipitated the glycoprotein IIb-glycoprotein IIIa (GPIIb-GPIIIa) complex from the surface of detergent solubilized intact human platelets; and it precipitated GPIIIa from the surface of chymotrypsin-treated platelets . Partially purified GPIIIa was also immunoprecipitated by MA 123 . Our data suggest that the exposure of fibrinogen receptors by ADP, chymotrypsin or pronase, is associated with alterations of GPIIIa on the platelet surface. J Antimicrob Chemother, 1984 Apr, 13(4), 347 - 52 Analysis of plasmids mediating gentamicin resistance in methicillin-resistant Staphylococcus aureus; Townsend DE et al.; Gentamicin-resistance plasmids in methicillin-resistant Staphylococcus aureus isolated from four Australian hospitals have been studied . All the plasmids conferred resistance to gentamicin, tobramycin, kanamycin and all but one conferred resistance to quarternary ammonium compounds . Plasmids which only carried these resistance determinants were ca 15.3 megadaltons and were only found in isolates from one hospital . The most common plasmid was ca 18.0 megadaltons and in addition encoded trimethoprim resistance . Two plasmids, one of ca 19.6 megadaltons and one of ca 28.5 megadaltons were found to carry the penicillinase determinant . However only the larger of these encoded heavy metal ion resistance and was sensitive to quaternary ammonium compounds . EcoR1 analysis indicated that all but the ca 28.5 megadalton plasmid were closely related . The EcoR1 analysis of the ca 28.5 megadalton plasmid indicated that it could have resulted from recombination between a gentamicin resistance determinant and a penicillinase plasmid. J Clin Invest, 1984 Apr, 73(4), 987 - 91 Antibody-mediated bacterial adhesion to cytomegalovirus-induced Fc receptors . Potential relationship to secondary infections complicating herpesvirus infections; Mackowiak PA et al.; Cytomegalovirus (CMV) and other viruses within the herpes group have recently been shown to induce Fc receptors in infected monolayers . We have examined the possibility that such receptors might facilitate the adherence of antibody-coated bacteria to CMV-infected cells . To do this, we infected confluent human embryonic lung (HEL) cell monolayers with CMV (strain AD 169) and then used a double radiolabel assay to measure adherence of Escherichia coli 06 to both infected and control monolayers . We examined infected monolayers 48 h after viral seeding, at which time 30-60% of the cells exhibited characteristic cytopathic changes . We compared the adherence of untreated E . coli 06 with the adherence of E . coli 06 that had been preincubated for 1 h at 37 degrees C with either nonimmune or anti-E . coli 06 antiserum . Pretreatment of the E . coli 06 with specific antiserum significantly enhanced its adherence to CMV-infected, but not to control, monolayers (P less than 0.01 by the Mann-Whitney U test) . We did not see such enhancement when we used anti-E . coli 06 antiserum to treat a nontypable E . coli . The augmented adherence of antibody-coated E . coli 06 to CMV-infected monolayers was abrogated by pretreating the monolayers with nonimmune serum or purified Fc fragments, but not by pretreating with IgA, IgM, or 1 mM trypan blue . Preincubating HEL cell monolayers with 100 U/ml human leukocyte interferon for 72 h at 37 degrees C did not affect the adherence of antibody-coated E . coli 06 to the monolayers . To determine if antibody-coated bacteria that adhered to the surface of CMV-infected monolayers might themselves act as receptors for microorganisms with Fc binding potential, we compared the adherence of Cowan strain Staphylococcus aureus to CMV-infected and control monolayers that had been preincubated with antibody-coated E . coli 06 . The S . aureus adhered significantly better to the former monolayers (P less than 0.001) . These results illustrate a previously unrecognized mechanism by which certain herpesviruses might enhance the adherence of secondary pathogens to nonphagocytic cell populations . Such a mechanism, if active in vivo, might facilitate the colonization of mucosal surfaces by these pathogenic microorganisms, and in this way might contribute to both the reported predisposition of CMV-infected patients to secondary infections and to the high prevalence of S . aureus in the vaginal flora of women with histories of genital herpes. J Clin Invest, 1984 Apr, 73(4), 1191 - 200 Pathogenesis of foreign body infection . Evidence for a local granulocyte defect; Zimmerli W et al.; Implanted foreign bodies are highly susceptible to pyogenic infections and represent a major problem in modern medicine . In an effort to understand the pathogenesis of these infections, we studied the phagocytic function in the vicinity of a foreign body by using a recently developed guinea pig model of Teflon tissue cages subcutaneously implanted (Zimmerli, W., F.A . Waldvogel, P . Vaudaux, and U.E . Nydegger, 1982, J . Infect . Dis., 146:487-497) . Polymorphonuclear leukocytes (PMN) purified from tissue cage fluid had poor bactericidal activity against a catalase-positive microorganism . When compared with blood or exudate PMN, they exhibited a significant reduction in their ability to generate superoxide in response to a particulate or a soluble stimulus (72 and 57%, respectively, P less than 0.001) . Not only their total contents in myeloperoxidase, beta-glucuronidase, lysozyme, and B12 binding protein were significantly reduced (by 62, 21, 47, and 63%, respectively, P less than 0.01), but also their capability for further secretion of residual B12 binding protein upon stimulation . Ingestion rates of endotoxin-coated opsonized oil particles were reduced by 25% (P less than 0.05) . In an effort to reproduce these abnormalities in vitro, fresh peritoneal exudate PMN were incubated with Teflon fibers in the presence of plasma . Interaction of PMN with the fibers led to significant increases in hexose monophosphate shunt activity and exocytosis of secondary granules (P less than 0.01) . PMN eluted after such interaction showed defective bactericidal activity, oxidative metabolism, and granular enzyme content similar to those observed in tissue cage PMN . The local injection of fresh blood PMN into tissue cages at the time of, or 3 h after, inoculation with 100 microorganisms (Staphylococcus aureus Wood 46) reduced the infection rate from 50 to 56 cages to 1 of 21 (P less than 0.001) and 3 of 8 cages (P less than 0.001), respectively . These results suggest that the in vivo as well as in vitro interaction of PMN with a nonphagocytosable foreign body induces a complex PMN defect, which may be partly responsible for the high susceptibility to infection of foreign bodies. Diagn Microbiol Infect Dis, 1984 Apr, 2(2), 85 - 91 Predominance of two newly described capsular polysaccharide types among clinical isolates of Staphylococcus aureus; Arbeit RD et al.; A capsular polysaccharide typing schema for Staphylococcus aureus, based upon the preparation of rabbit typing sera with eight prototype strains, has been reported ( Karakawa and Vann , 1982) . These antisera were used to classify the capsular polysaccharides of 246 S . aureus isolates from patients in a survey of hospitals in several countries and 49 consecutive blood isolates obtained over a 17-month period in a clinical study at the Boston Veterans' Administration Medical Center . Two capsular types, 5 and 8, accounted for about 70% of these isolates; most of the remaining strains could not be typed with the available antisera . The clinical study of bacteremia identified capsular types 5 and 8 among both community-acquired and nosocomial isolates and showed that strains bearing these two types caused the patterns of disease reported for staphylococcal bacteremia . There was an association between the phage type and the capsular type of these bacteremic strains . The capsular types of the "classic" encapsulated strains of S . aureus, M (type 1) and Smith (type 2), were not observed among blood isolates in this study . The observation that most clinical isolates of S . aureus belong to two recently defined capsular types provides a new focus for investigations into the virulence of this common nosocomial pathogen and suggests the potential for protective acquired immunity against staphylococcal bacteremia. J Clin Microbiol, 1984 Apr, 19(4), 453 - 6 Fluorescent staining of intracellular and extracellular bacteria in blood; Mansour JD et al.; The fluorescent dye ethidium bromide stains Escherichia coli and Staphylococcus aureus in whole blood . The staining is rapid, relatively specific, and does not require fixation of the sample . Furthermore, stained bacteria can be seen microscopically without the need for a final wash to remove unbound dye . By using lysostaphin, an S . aureus-specific lytic enzyme, we have demonstrated that S . aureus can be stained with ethidium bromide even after phagocytosis . After short periods of incubation with the dye (less than 5 min), bacteria, both intracellular and extracellular, were the predominant fluorescent particles . With increasing time of incubation, blood cell components, notably leukocyte nuclei, began to fluoresce. Infect Immun, 1984 Apr, 44(1), 175 - 81 Purification and characterization of Staphylococcus aureus FRI 1169 and 587 toxic shock syndrome exotoxins; Igarashi H et al.; An exotoxin was purified from a toxic shock toxin (TST)-producing Staphylococcus aureus strain, FRI 1169, and another exotoxin was purified from a pyrogenic exotoxin C (PEC)-producing S . aureus strain, 587 . Both strains had been isolated from toxic-shock syndrome patients . The two exotoxins were purified by the same method of ion-exchange chromatography, chromatofocusing, and gel filtration . After purification, those exotoxins gave a line of identity against an anti-TST serum and also were immunologically similar to TST in a double-diffusion test . In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, each exotoxin gave a single band with a relative mobility identical to that of the other . Their molecular weights (24,000), isoelectric points (7.0), amino acid compositions, and NH2-terminal amino acid sequences (the first four residues) were identical . Both produced fever and enhanced host susceptibility to lethal endotoxin shock in rabbits, comparable with PEC . These findings show that the two exotoxins are the same protein, which is assumed to be TST . When injected into rabbits, the culture supernatant of strain 587 showed biological activity like that described above, whereas the culture supernatant neutralized with anti-TST immunoglobulin did not . This showed that PEC-producing strain 587 does not produce any toxin with these biological activities in rabbits except TST. J Immunol, 1984 Apr, 132(4), 1761 - 6 Deoxyadenosine modulates human suppressor T cell function and B cell differentiation stimulated by Staphylococcus aureus protein A; Cohen AH et al.; Adenosine deaminase (ADA) deficiency and the resultant accumulation of deoxyadenosine (AdR) are associated with profound T cell dysfunction and variable B cell dysfunction in vivo . We examined the effects of AdR on the in vitro function of normal human peripheral blood B and T lymphocytes whose ADA activity was inhibited by 2'-deoxycoformycin . We found that OKT8+ T cell-mediated suppression of SPA-induced Ig production was markedly reduced by concentrations of AdR (3 to 10 microM) that did not affect helper T cell function . Because the lectin-induced proliferative responses of OKT8+ T cells and OKT8- T cells were equally susceptible to AdR, modulation of in vitro immune responses by low-dose AdR probably reflected different proliferative requirements for the expression of T cell helper or suppressor functions . Although low doses of AdR did not inhibit Ig production in SPA-stimulated cultures, we found that T cell-dependent, SPA-stimulated B cell proliferation was blocked by 3 to 10 microM AdR . Therefore, it appeared that B cell proliferation was not required for the induction of Ig synthesis in this system . Higher doses (30 to 100 microM) of AdR did block the induction of Ig synthesis, presumably by interfering with T-helper functions via a mechanism other than inhibition of proliferation and/or by inhibiting B cell differentiation events. Biochim Biophys Acta, 1984 Mar 29, 785(3), 104 - 10 The reversible deactivation of beta-lactamase from Staphylococcus aureus by quinacillin and cephaloridine and its modification by antibodies; Carrey EA et al.; The effect of antibody on the reversible deactivation of the beta-lactamase (penicillin amino-beta-lactamhydrolase, EC 3.5.2.6) from Staphylococcus aureus has been studied using quinacillin and cephaloridine as substrates . The latter has been shown to exhibit the characteristics of an A-type substrate Citri, N., Samuni, A . and Zyk, N . (1976) Proc . Natl . Acad . Sci . U.S.A . 73, 1048-1052) and reversibly to lower the activity of the enzyme towards benzylpenicillin in a manner analogous to quinacillin . Both divalent and monovalent antibodies reduce the activity of the lactamase to 60% of the native value in the absence of substrate . The reduction by monovalent antibody is slow (t1/2 approximately equal to 25 min) . Both divalent and monovalent antibodies modify the time-course of reversible deactivation independently of being added before or subsequent to deactivation by substrate . The full recovery of activity is delayed in the case of quinacillin and accelerated for cephaloridine . The activity against benzylpenicillin in the deactivated states is unaffected . These effects are shown to reflect the changed rates of hydrolysis of the two substrates in the presence of antibody . The effect of antibody is mediated by minor conformational change . Continuous assays for following the hydrolysis of quinacillin and cephaloridine by optical rotation are reported. J Biol Chem, 1984 Mar 10, 259(5), 3152 - 9 Induction of vascular smooth muscle alpha-isoactin expression in BC3H1 cells; Strauch AR et al.; An isoactin analysis was performed on L-{35S}cysteine labeled BC3H1 cells to determine if these smooth muscle-like cells synthesize vascular smooth muscle actin . Three different NH2-terminal peptides were identified on thin layer electrophoretograms of DNase I-purified and trypsin-digested BC3H1 cell actin . Results obtained from secondary digestion with thermolysin or Staphylococcus aureus V8 protease showed that the most acidic NH2-terminal peptide was derived from vascular smooth muscle alpha-isoactin . Treatment of cell monolayers with serum-free medium caused a 3-fold increase in the level of alpha-isoactin expression and a concomitant decrease in the level of non-muscle beta- and gamma-isoactin . Cell-cell contact was required for induction of alpha-isoactin, and the effects of serum depletion on isoactin expression and cell growth were reversible . The intensity of about 11 out of 500 polypeptide spots on two-dimensional gels of BC3H1 cell polypeptides also was influenced by the culture conditions . The finding that smooth muscle isoactin expression was coupled to cell growth conditions indicate the potential usefulness of BC3H1 cells in studies of isoactin expression and utilization during vascular smooth muscle development. J Biol Chem, 1984 Mar 10, 259(5), 3350 - 4 Association of alpha- and beta-subunits during the biosynthesis of beta-hexosaminidase in cultured human fibroblasts; Proia RL et al.; Subunit association of beta-hexosaminidase was studied in intact fibroblasts using antisera that discriminate between free and associated alpha-chains . These were anti-beta-hexosaminidase A (anti-alpha beta), which precipitated all alpha-chains, free or associated; anti-beta-hexosaminidase B (anti-beta beta), which precipitated those alpha-chains that were associated with beta; and anti-alpha-chains, which recognized only monomeric alpha-chains . After biosynthetic labeling, beta-hexosaminidase or its free alpha-subunit were immuno-precipitated from extracts of cells and medium with the aid of protein A-bearing Staphylococcus aureus, subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and visualized by fluorography . Pulse-chase labeling showed that the alpha-chains existed predominantly in the monomeric precursor form during the first 5 h, and then began to accumulate in the mature (lysosomal) associated alpha beta form . Precursor alpha beta complexes were secreted, along with some precursor alpha monomers; the latter were catalytically inert . Both alpha- and beta-chains were phosphorylated (a Golgi modification) prior to association . Thus alpha-beta association probably occurred in the Golgi area before transfer to lysosomes and before secretion . Cycloheximide inhibited the association and subsequent maturation of preformed alpha-chains, perhaps by causing a depletion of a pool of beta-chain precursor upstream from the site of subunit association . In fibroblasts from a patient with Sandhoff disease, that produced no beta-chains, the alpha-chains self-associated but their maturation was markedly decreased . We suggest that association with beta-chains is necessary not only for acquisition of catalytic activity but also for transport of alpha-chains to lysosomes. Biochim Biophys Acta, 1984 Mar 7, 792(3), 367 - 70 Improvement in the lipid extraction of staphylococcal cells by lysostaphin treatment; Rizzo AF et al.; A new method for extracting lipids from Staphylococcus aureus is described in this paper . The extracts of cells treated with lysostaphin are compared with those from untreated cells . The recovery of lipids improved with the enzymatic treatment . Tentative identification of the components of the lipid extracts has been carried out. Arch Fr Pediatr, 1984 Mar, 41(3), 201 - 3 {Staphylococcal toxic shock}; Guillois B et al.; The authors report the case of a 5 year-old child who, after a sore throat, developed gastrointestinal problems, high fever, scarlatinaform rash, conjunctivitis, shock with renal failure and involvement of liver, pancreas and muscles . No infectious site was detected . However, he had positive blood culture for staphylococcus aureus . The child fully recovered after a period of desquamation of the palms and soles. Arch Inst Cardiol Mex, 1984 Mar-Apr, 54(2), 137 - 44 {Right infectious endocarditis . Study of 30 cases}; Requeni F et al.; Due to the lack of specificity of the clinical picture in the right-sided infective endocarditis, the correct diagnosis is rarely made . We reviewed 30 cases with right-sided or right and left infective endocarditis, treated in the INC from 1946 to 1982 . The average age was 20 years . Rheumatic fever (53%), congenital heart disease (40%) and cardiac prostheses (7%) were the more common underlying diseases . The diagnosis was made on an average 7.3 months after the first symptom . Heart failure (93%), fever (76%), weight loss (73%), haemoptysis (66%) and general malaise (53%) were the predominant symptoms . There was no diagnostic suspicion in 9 patients (30%) and in 7 from 16 with negative blood culture, the infection was exclusively right-sided . Peripheral and pulmonary embolism was the most frequent complication . (66%) There were 29 deaths (96.6%) . In all of them the diagnosis was confirmed in the postmortem examination . Heart failure and septic shock were the main causes of death . Almost all patients were infected with gram-negative germs and staphylococcus Aureus . This diagnosis should be suspected in a patient with known heart disease, who develops unexplained heart failure, moreover if pulmonary emboli are a feature . The diversity of the isolated germs is different from other publication that have shown staphylococcus as the most prevalent microorganism . This difference can be explained by the lack of drug abuse in our cases . The mortality rate is higher than in the left sided endocarditis. Southeast Asian J Trop Med Public Health, 1984 Mar, 15(1), 63 - 7 Infection rates of respiratory syncytial virus in pediatric patients attending Phra Mongkutklao Hospital, Bangkok; Tantivanich S et al.; Respiratory syncytial virus (RSV) and other pathogens were isolated from nasopharyngeal secretions from 200 pediatric patients attending the Out Patient Department of Phra Mongkutklao Hospital with symptoms of upper respiratory tract infections . Their sera were also taken for determination of class specific immunoglobulin antibody titers . The positive isolation rates were 36% for RSV, 5.5% for adenovirus 1.5% for herpes simplex virus (HSV), and 4% for Staphylococcus aureus . One to 5.5% of these patients had mixed infection . Ninety five percent of patients with positive RSV isolations had IgM antibody which was found only in 30.7% in patients with negative RSV isolations . This result indicated that RSV was likely to be the most common pathogen responsible for the upper respiratory tract infections in children in Bangkok during the rainy season. Antibiotiki, 1984 Mar, 29(3), 201 - 5 {Fatty acid composition of variant Staphylococcus aureus 209 P resistant to antibiotic AL-87}; Smirnov VV et al.; The antibiotic AL-87 resistant variant of S . aureus 209P was grown on the antibiotic-free medium . The study on its fatty acid composition showed that it was analogous to the fatty acid composition of the initial sensitive strain with the predominance of branched fatty acids . However, when the resistant variant was grown on the antibiotic-containing medium, it was characterized by a markedly changed fatty acid composition with the predominance of the straight chain saturated and unsaturated fatty acids, mainly hexadecanoic and octadecenic acids . The fatty acid composition of the resistant variant grown in the presence of the antibiotic was close to that of a wide range of gram-positive and gram-negative microogranisms. Orig Life, 1984 Mar, 13(3-4), 169 - 76 The tubulins of animals, plants, fungi and protists implications for metazoan evolution; Little M et al.; alpha-Tubulin subunits from trout (S . gairdneri) sperm tails, sea urchin (S . purpuratus) cilia, protistan alga (C . elongatum ) flagella and rose (Paul's Scarlet) cytoplasm have been characterized by limited proteolytic cleavage with the enzyme Staphylococcus aureus protease and electrophoresis of the digestion products on SDS-PAGE . The resulting patterns corresponded to either of two major types representative of animal and non-animal alpha-tubulins, respectively . A total of 28 alpha-tubulins have now been characterized by this method . They are classified in this paper according to the type of cleavage pattern generated by the enzyme S . aureus protease . The implications of these results for metazoan evolution are discussed. J Dairy Sci, 1984 Mar, 67(3), 620 - 7 Staphylococcal capsular vaccine for preventing mastitis in two herds in Georgia; Yoshida K et al.; An encapsulated staphylococcal vaccine, consisting of heat-killed capsular-type A and B Staphylococcus aureus strains and capsular polysaccharide extracted from strain ATCC 31432 of Staphylococcus epidermidis, was used to control bovine mastitis in two herds in Georgia . The vaccine was administered intramuscularly into 97 and 125 cows in the herds, and equal numbers of animals were controls . Two weeks after primary vaccination a booster injection was given . No side effects were observed . In one herd, leucocyte content of milk samples decreased remarkably 1 wk after the booster injection . Significant resistance to infection was maintained for 4 mo after vaccination . Estimation of the total loss of milk yield showed less loss compared to that in the control group for 4 mo after vaccination . In the other herd, remarkable improvements of milk samples were observed as early as 1 wk after primary vaccination and resistance to infection continued for 6 mo after vaccination, when experiments were terminated . At 3 mo after vaccination, loss of milk yield was approximately one-third of that in the control group, and this reduction of loss was maintained for 6 mo after vaccination. Am J Vet Res, 1984 Mar, 45(3), 420 - 3 Effect on outcome of intramammary challenge exposure with Staphylococcus aureus of somatic cell concentration and presence of an intramammary device; Schultze WD et al.; Results of experimental Staphylococcus aureus intramammary challenge of all quarters of 6 cows, each fitted with an intramammary device (IMD) in 2 quarters, and of 10 quarters of 3 cows not fitted with an IMD were reported . Infection was established in all 34 quarters, regardless of presence or absence of an IMD . Neither the course nor severity of early S aureus intramammary infection were influenced by the presence of an IMD or by differences in milk somatic cell (MSC) concentration in the gland at the time of bacterial challenge infusion, up to a MSC concentration of nearly 1 million/ml . Cumulative success of experimental infection in this and a previous study from our laboratory was nearly 100% in glands in which the MSC concentration was less than 1 million/ml and about 17% when the MSC concentration exceeded 1 million/ml. J Trauma, 1984 Mar, 24(3), 208 - 13 The role of plasma fibronectin as a nonantibody, noncomplement opsonin for Staphylococcus aureus; Deitch EA et al.; The possible role of plasma fibronectin as a nonantibody, noncomplement opsonin for S . aureus was studied using peripheral blood leukocytes from healthy rabbits . Fibronectin depletion of normal rabbit serum by affinity absorption chromatography reduced the opsonic ability of that serum (p less than 0.05) and resulted in impaired bacterial killing of S . aureus in vitro . The addition of purified fibronectin to fibronectin-depleted serum significantly reversed the opsonic defect (p less than 0.01) . The combination of complement inactivation, plus fibronectin depletion, resulted in a severe opsonic deficiency that was much worse than either deficiency alone (p less than 0.05) . However, fibronectin alone in the absence of other serum factors was a poor opsonin for S . aureus, suggesting that its major role as an opsonin might be to augment or amplify other serum factors. Arch Ophthalmol, 1984 Mar, 102(3), 461 - 3 Occurrence of phlyctenules after immunization with ribitol teichoic acid of Staphylococcus aureus; Mondino BJ et al.; We immunized rabbits to ribitol teichoic acid (RTA), a component of the cell wall of Staphylococcus aureus, by intravenous injections of RTA-sensitized sheep RBCs . Hemagglutination titers to RTA and corneal phlyctenules developed in these rabbits after topical challenge with viable S aureus, but did not develop in control rabbits immunized with unsensitized sheep RBCs . These results suggest that hypersensitivity to RTA, the major antigenic determinant of S aureus, plays a role in the development of corneal phlyctenules. Arch Intern Med, 1984 Mar, 144(3), 541 - 5 Complications associated with Staphylococcus aureus bacteremia; Libman H et al.; Thirty-nine consecutive Staphylococcus aureus bacteremias were reviewed with particular attention to complications . Thirty-four (87%) of the bacteremias were nosocomial, with intravascular catheters (20 episodes) and dialysis-access sites (six episodes) the most common sources . Complications developed in 36% (14/39) of all bacteremias and in 30% (6/20) of those that were catheter-associated . Acute complications (shock, adult respiratory distress syndrome, disseminated intravascular coagulation) occurred in six patients and were fatal in four . In nine patients metastatic suppurative complications developed, six at sites of preexisting abnormalities . There were no episodes of endocarditis . Most patients received prolonged antibiotic therapy, and the majority of all suppurative complications required surgical intervention . Staphylococcus aureus bacteremia, even when not associated with endocarditis, is a cause of considerable morbidity and mortality in hospitalized patients. South Med J, 1984 Mar, 77(3), 302 - 3 Abuse of antibiotics by abusers of parenteral heroin or cocaine; Novick DM et al.; We studied antibiotic intake in 197 abusers of alcohol, sedatives, or parenteral heroin or cocaine . Thirteen patients, all abusers of parenteral heroin or cocaine, had taken antibiotics without prescription, obtained from friends, from old prescriptions, or by purchase on the street . Past or present street purchase of antibiotics was noted in 27 patients and was more common (P less than .02) in parenteral substance abusers . Four parenteral substance abusers had infection or colonization with methicillin-resistant Staphylococcus aureus, and three of them had purchased antibiotics on the street before the present or a recent past hospitalization . Physicians treating abusers of parenteral heroin or cocaine should be aware that such patients may be taking antibiotics without medical supervision . This practice may be an important factor in the development and spread of methicillin-resistant S aureus. J Surg Res, 1984 Mar, 36(3), 237 - 43 The biochemical bonding of cefoxitin to a microporous polytetrafluoroethylene surface; Greco RS et al.; Cefoxitin was bound to a microporous polytetrafluoroethylene (PTFE) surface with tridodecylmethylammonium chloride (TDMAC) . Bactericidal concentrations of cefoxitin were achieved with very small doses of the antibiotic . Elution of cefoxitin from microporous PTFE occurs by two concurrent first-order processes, each occurring at a different rate constant . Bound cefoxitin inhibits the growth of Staphylococcus aureus in a bioassay . Finally, cefoxitin can be adsorbed to TDMAC-treated microporous PTFE in vivo when the antibiotic is administered locally or intravenously . The application of antibiotic bonding to the prevention of vascular prosthetic infections is discussed. J Pediatr Orthop, 1984 Mar, 4(2), 162 - 9 Foci of chronic circumscribed osteomyelitis (Brodie's abscess) that traverse the epiphyseal plate; Bogoch E et al.; We observed six children who presented with chronic circumscribed osteomyelitis involving the adjacent metaphysis and epiphysis of a long bone, communicating through and damaging the growth cartilage of the epiphyseal plate . Four of the six children were less than or equal to 10 years of age . All six patients presented with the mild symptoms and subtle clinical findings that are characteristic of "Brodie's abscess," which is usually confined to metaphyseal, or occasionally epiphyseal, bone . Four children were treated with antibiotics and by surgical evacuation of the abscess, with visualization of the defect in the epiphyseal plate . Two children were treated with antibiotics alone, initially by the intravenous route . At follow-up 2-14 years after treatment, all affected children had a normal result without evidence of growth disturbance . There are seven previously reported cases of chronic circumscribed osteomyelitis traversing the epiphyseal plate that resulted in growth disturbance . Based on our experience and that reported in the literature, we believe that the intravenous administration of appropriate antibiotics in high doses, followed by oral antibiotics, is sufficient treatment for some children presenting with this condition . The pathogenic organism is likely to be Staphylococcus aureus . Surgical evacuation of the lesion should be performed for acute osteomyelitis involving the epiphyseal plate, for sinus formation or drainage into a synovial joint, for failure of the patient to respond clinically to nonoperative therapy, and for confirmation of the diagnosis if doubt exists. J Bacteriol, 1984 Mar, 157(3), 863 - 7 Quantitative association between electrical potential across the cytoplasmic membrane and early gentamicin uptake and killing in Staphylococcus aureus; Eisenberg ES et al.; The relationship between the magnitude of the transmembrane electrical potential and the uptake of {14C}gentamicin was examined in wild-type Staphylococcus aureus in the logarithmic phase of growth . The electrical potential (delta psi) and the pH gradient across the cell membrane were determined by measuring the equilibrium distribution of {3H}tetraphenyl-phosphonium and {14C}acetylsalicylic acid, respectively . Incubation in the presence of the H+-ATPase inhibitor N,N'-dicyclohexylcarbodiimide (DCCD) led to an increase in delta psi with no measurable effect on the pH gradient at external pHs ranging from 5.0 to 6.5, and the effect on delta psi was DCCD concentration dependent . In separate experiments, gentamicin uptake and killing were studied in the same cells under identical conditions . At pH 5.0 (delta psi = -140 mV), no gentamicin uptake occurred . In the presence of 40 and 100 microM DCCD, delta psi was increased to -162 and -184 mV, respectively, and gentamicin uptake was observed in a manner that was also dependent on the DCCD concentration . At pH 6.0 (delta psi = -164 mV), gentamicin uptake occurred in the absence of the carbodiimide but was enhanced in a concentration-dependent fashion by 40 and 100 microM DCCD (delta psi = -174 and -216 mV, respectively) . In all cases increased gentamicin uptake was associated with an enhanced bactericidal effect . The results indicate that initiation of gentamicin uptake requires a threshold level of delta psi (-155 mV) and that above this level drug uptake is directly dependent on the magnitude of delta psi. Infect Immun, 1984 Mar, 43(3), 954 - 8 Association of high levels of serum antibody to staphylococcal toxic shock antigen with nasal carriage of toxic shock antigen-producing strains of Staphylococcus aureus; Ritz HL et al.; Forty-four asymptomatic male subjects were examined for their nasal carriage of strains of Staphylococcus aureus capable of producing staphylococcal toxic shock antigen (TSA), an exotoxin implicated in the pathogenesis of toxic shock syndrome . In addition, the levels of antibody to TSA in sera from these subjects were determined by an enzyme-linked immunosorbent assay . S . aureus was isolated from the anterior nares of 23 subjects . Of those 23 isolates of S . aureus, 9 were found to produce TSA . All individuals carrying strains of S . aureus capable of producing TSA had high to moderate levels of antibody to TSA . In contrast, those individuals carrying strains not producing TSA had levels of antibody to TSA ranging from high to nondetectable . A second examination of nasal samples from 42 of these subjects revealed that 86% of those carrying S . aureus initially still carried S . aureus after a period of 3 months; all subjects found to carry TSA-producing strains initially and that were examined a second time yielded TSA-producing strains once again. Eur J Biochem, 1984 Mar 1, 139(2), 235 - 46 Multiple forms of hepatic cytochrome P-450 . Purification, characterisation and comparison of a novel clofibrate-induced isozyme with other major forms of cytochrome P-450; Tamburini PP et al.; In the present studies, a novel form of highly purified cytochrome P-450 (cytochrome P-452) isolated from the hepatic microsomes of clofibrate-pretreated rats has been compared to the major isozymes isolated from the hepatic microsomes of rats pretreated with phenobarbital (cytochrome P-450) and 2-naphthoflavone (cytochrome P-447) using a number of biochemical criteria . The results show that these three isozymes exhibit marked structural differences from each other as judged by a complete lack of immunochemical cross-reactivity between the isozymes and the heterologous rabbit serum antibodies using Ouchterlony double diffusion, and non-identity between the limited proteolytic digestion maps of the three isozymes obtained in the presence of chymotrypsin, papain and Staphylococcus aureus V8 proteases . Furthermore, the three isozymes exhibited clear differences in their monomeric molecular weights determined on calibrated sodium dodecyl sulphate/polyacrylamide gel electrophoresis in gels of varying acrylamide concentration . Substantial differences were also observed in the substrate specificities of the isozymes, which were reflected in differences in the turnover rates and positional selectivities of the hemoproteins for some model substrates . In addition, the isozymes differed in their substrate binding affinities and their ability to interact with purified hepatic microsomal cytochrome b5, as judged using difference spectrophotometry . Finally, subtle differences were detected in the ultraviolet visible absorbance spectra of the hemoproteins in the ferric, ferrous, and carbonmonoxyferrous states . Taken collectively, the above data provides compelling evidence that fundamental differences exist between these cytochrome P-450 isozymes, further establishing the uniqueness of the major form of cytochrome P-450 induced by clofibrate pretreatment. Exp Cell Res, 1984 Mar, 151(1), 264 - 7 Changes in deoxyadenosine production during human lymphocyte mitogenesis; Iizasa T et al.; Immunodeficient children who